Calcareous Compacted Mine Soil in Southeast Ohio

Calcareous Compacted Mine Soil in Southeast Ohio: A Prairie Grass Habitat

Dissertation

Presented in Partial Fulfillment of the Requirements for the Degree Doctor of

Philosophy in the Graduate School of The Ohio State University

By

Mark Ervin Thorne, B.S., M.S.

Environmental Science Graduate Program

The Ohio State University

2010

Dissertation Committee:

John Cardina, Advisor

James Metzger

Landon Rhodes

Craig Davis

Copyright by

Mark Ervin Thorne

2010

ii

Abstract

The degradation of terrestrial ecosystems has been an unfortunate by-product

of the human demand for natural resources. As a result, environmental quality suffers

as watersheds are lost, global warming continues, species go extinct, and productivity

is reduced. Restoring ecosystems such as prairies or forests to their original character

is desirable but may be impossible if the extent of degradation is too great.

Rehabilitation of severely degraded ecosystems may require establishment of

alternative communities to recreate functional ecosystem attributes such as diversity,

stability, and productivity. In Ohio, surface coal mining has replaced thousands of

hectares of diverse forests with a low-diversity, non-native forage complex. Adverse

soil conditions have prohibited reestablishment of native forest species; thus,

restoration of the original ecosystem is unlikely. As an alternative, establishment of a

diverse prairie would restore ecosystem attributes and be more desirable than the

current forage complex. This research examines suitability of native prairie grasses on

reclaimed mine land in southeast Ohio. Grass species were selected form habitats

across North America and represent differences in climate and native habitat. Grasses

were planted on compacted, calcareous, bare-soil field plots in 2005 and 2006 on land

that had been mined and reclaimed in the mid-1980s and was previously supporting a

low-diversity, non-native forage complex. Establishment and growth of prairie

grasses was monitored through 2007 with population counts, cover assessments, and

iii

biomass measurements. Glasshouse experiments were conducted to compare growth

of selected grasses and effectiveness of mine soil arbuscular-mycorrhizal fungi (AMF)

with native tallgrass prairie AMF at different levels of soil phosphorus and

compaction. Eastern gamagrass (Tripsacum dactyloides L.) was the most successful

in relation to number of seeds sown and equaled productivity of the existing

vegetation by the second year. Eastern gamagrass was also found to be highly

dependent on AMF when phosphorus was low. Tall dropseed (Sporobolus asper

(Michx.) Kunth) was successful and tolerated low phosphorus without AMF.

Buffalograss (Buchloe dactyloides (Nutt.) Engelm) established well with spreading

stolons and produced a dense ground cover within the first year of growth. Big

bluestem (Andropogon gerardii Vitman), Indiangrass (Sorghastrum nutans (L.) Nash),

and little bluestem (Schizachyrium scoparium (Michx.) Nash) are highly dependent on

AMF and were least successful in establishment, but increased in productivity during

the study. Sideoats grama (Bouteloua curtipendula (Michx.) A. Gray) and blue grama

(Bouteloua gracilis (Willd. ex Kunth) Lag. ex Griffiths) did not persist, likely due to

periodic anaerobic soil. Mine soil AMF were found to be equally or more effective

than the tallgrass prairie AMF, and would likely not be a barrier for establishment of

other prairie species. Soil compaction was not a factor for eastern gamagrass but early

growth was greater for tall fescue (Festuca arundinaceae Schreb.), one of the existing

non-native forage species, which explains its success in mine soil revegetation.

Establishing a diverse prairie community on mine soil can likely be accomplished, but

with a mix of prairie species that contain different strategies to overcome the poor

growing conditions of the mine soil habitat.

iv

Dedication

I dedicate this work to my best friend,

Deborah Mills Thorne.

The journey, we share.

v

Acknowledgements

This work would not have occurred without the outstanding support and

mentoring from Dr. John Cardina, my advisor, who was genuinely interested in my

success. I also sincerely appreciate the members of my committee, Dr. James

Metzger, Dr. Landon Rhodes, and Dr. Craig Davis who provided advice, laboratory

space and equipment, and were great mentors in the classroom. I recognize and

appreciate the assistance of Dr. Nicole Cavender of the Wilds, who provided a great

field location for this research. Assistance given by Jim Vent and David Snodgrass at

the Howlett greenhouse and nursery on The Ohio State University campus was

invaluable. First-year funding provided by Dr. Martin Quigley, and consultation given

by Dr. J.P. Vimmerstedt, Professor Emeritus, The Ohio State University / Ohio

Agricultural Research and Development Center, and Gary Kaster, American Electrical

Power, was also greatly appreciated. And, I am forever grateful to The Ohio State

University for awarding me a Mary S. Muellhaupt Presidential Fellowship, which

significantly aided in the completion of this work.

vi

Vita

1980 ..................... A.A. Liberal Arts, Blue Mountain Community College, Pendleton,

OR.

1989 ..................... B.S. Crop Science, Oregon State University, Corvallis, OR.

1990-1994 ........... Agricultural Research Technician, Washington State University,

Pullman, WA.

1996 ..................... M.S. Natural Resource Sciences, Washington State University,

Pullman, WA.

1996-1997 ........... Associate of Research, Washington State University, Pullman, WA.

1997-1998 ........... Agricultural Research Technologist, Washington State University,

Pullman, WA.

2000 ..................... Instructor – Crop Science 305 – Principles of Weed Science,

Washington State University. Pullman, WA.

2002 ..................... Graduate Teaching Associate – Weed Science 422 – Principles of

Weed Science, The Ohio State University, Columbus, OH.

2004 ..................... Graduate Teaching Associate – Environment and Natural Resources

201 – Introduction to Environmental Science, The Ohio State

University, Columbus, OH.





203 – Society and Natural Resources, The Ohio State University,

Columbus, OH.

2007 ..................... Mary S. Muellhaupt Presidential Fellowship, The Ohio State


vii




Publications

1990 ..................... Eastern Washington Weed Control Report, C.M. Boerboom, Dept.

of Crop and Soil Sciences, Washington State University, Pullman,

WA.

1991 ..................... Eastern Washington Weed Control Report, C.M. Boerboom, and

M.E. Thorne, Dept. of Crop and Soil Sciences, Washington State

University, Pullman, WA.


M.E. Thorne, Technical Report 93-1, Dept. of Crop and Soil

Sciences, Washington State University, Pullman, WA.


M.E. Thorne, Technical Report 94-3, Dept. of Crop and Soil

Sciences, Washington State University, Pullman, WA.

1998 ..................... Thorne, M.E., B.A. Zamora, and A.C. Kennedy. Sewage sludge

and mycorrhizal effects on Secar bluebunch wheatgrass in mine

spoil. Journal of Environmental Quality 27:1228-1233.

1998 ..................... Integrated spring cropping systems for the semiarid wheat-fallow

region. Thorne, M. and F. Young. Technical Report 98-1, Dept. of

Crop and Soil Sciences, Washington State University, Pullman,

WA.

2003 ..................... Thorne, M.E., F.L. Young, W.L. Pan, R. Bafus, and J.R. Alldredge.

No-till spring cereal cropping systems reduce wind erosion

susceptibility in the wheat/fallow region of the Pacific Northwest.

Journal of Soil and Water Conservation 58:250-257.

2004 ..................... Thorne, D., A. Tickamyer, and M. Thorne. Poverty and income in

Appalachia. Journal of Appalachian Studies, Special Issue:

Appalachian Counts: The Region in the 2000 Census. 10:341-358.

viii

2004 ..................... Young, F.L. and M.E. Thorne. Weed-species dynamics and

management in no-till and reduced-till fallow cropping systems for

the semi-arid agricultural region of the Pacific Northwest, USA.

Crop Protection 23:1097-1110.

2006 ..................... Young, F.L., M.E. Thorne, and D.L. Young. Nitrogen fertility and

weed management critical for continuous no-till wheat in the

Pacific Northwest. Weed Technology 20:658-669.

2007 ..................... Thorne, M.E., F.L. Young, J.P. Yenish. Cropping systems alter

weed seed banks in Pacific Northwest semi-arid wheat region.

Crop Protection 26:1121-1134.

Fields of Study

Major Field: Environmental Science Graduate Program

ix

Table of Contents

Abstract ......................................................................................................................... ii

Dedication .................................................................................................................... iv

Acknowledgments ......................................................................................................... v

Vita ............................................................................................................................... vi

List of Tables................................................................................................................ xi

List of Figures ............................................................................................................. xv

Chapter 1: Introduction ................................................................................................ 1

Chapter 2: Prairie Grass Establishment on Calcareous Reclaimed Mine Soil ............. 4

Chapter 3: Productivity of Native Prairie Grasses on Reclaimed Calcareous

Mine Soil ............................................................................................................... 44

Chapter 4: Safe-sites Control Warm-season Prairie Grass Establishment on

Calcareous Compacted Mine Soil ............................................................................... 70

Chapter 5: Effectivity of Arbuscular Mycorrhizal Fungi of Reclaimed Calcareous

Surface Mine Soil and Native Tallgrass Prairie Soil on Growth of

Prairie Grasses ....................................................................................................... 87

x

Chapter 6: Effect of soil compaction and arbuscular-mycorrhizal fungi on tall fescue,

big bluestem and eastern gamagrass seedling growth......................................... 112

Chapter 7: Summary and Conclusions ..................................................................... 134

References ................................................................................................................. 138

Appendix A: Photographs of Reclaimed Surface Mined Land in Southeast Ohio .. 160

Appendix B: Cover Measurements of Existing Mine Soil Vegetation Re-establishing

in Prairie Grass Plots at the Conclusion of the Research .................................... 165

xi

List of Tables

Table 1. List of grasses planted in this research and a description of habitat

characteristics where each species is native. ......................................................... 32

Table 2. Precipitation data recorded 17 km northwest of the Wilds study site at the

Zanesville, Ohio municipal airport. ...................................................................... 35

Table 3. Soil analysis of reclaimed calcareous mine soil and adjacent non-mined forest

soil collected from a former surface coal mine in southeast Ohio. ....................... 36

Table 4. Properties of soil collected from the top 20 cm, in 5-cm increments, from

reclaimed coal mined study site in southeast Ohio. Samples collected randomly

from the Wilds study site on October 2007. ......................................................... 37

Table 5. Population density 30 days after planting (30DAP) in 2005 and 2006 in

relation to number of viable seeds sown. .............................................................. 38

Table 6. Changes in population density measured 30 days after planting (30DAP), in

October of the first year following planting (YR1), in October of the second and

third year after sowing (YR2 and YR3), respectively........................................... 39

Table 7. Species comparison of stand density as a percent of viable seeds sown, at

each census for the 2005 and 2006 plantings. ....................................................... 41

xii

Table 8. Properties of soil collected from the top 20 cm, in 5 cm increments, from

reclaimed coal mined study site in southeast Ohio. Samples collected randomly

from the Wilds study site on October 2007. ......................................................... 66

Table 9. Canopy cover of grasses, measured the first (YR1), second (YR2), and third

(YR3) years following planting, on reclaimed mine soil in southeast Ohio. ........ 67

Table 10. Aboveground biomass of grasses, measured in censuses in October of the

first (YR1), second (YR2), and third (YR3) years following planting, on reclaimed

mine soil in southeast Ohio. .................................................................................. 68

Table 11. Per-plant biomass of grasses growing on calcareous reclaimed mine soil in

southeast Ohio measured the year of planting (YR1) and in the second (YR2) and

third (YR3) years following planting. ................................................................... 69

Table 12. Effect of seeding rate on stand establishment of three warm season native

grasses on reclaimed calcareous mine spoil. ......................................................... 84

Table 13. Effect of seeding rate on stand biomass of three warm season native grasses

on reclaimed calcareous mine spoil. ..................................................................... 85

Table 14. Soil properties of reclaimed mine and AMF inoculum soil used to compare

growth of prairie grasses with different concentrations of phosphorus (P) and

different sources of arbuscular mycorrhizal fungi (AMF). ................................. 105

Table 15. Percent colonization by arbuscular mycorrhizal fungi (AMF) as affected by

AMF sources, soil phosphorus concentration (P), and grass species (SPP) in a

16-week glasshouse experiment. ......................................................................... 106

xiii

Table 16. Biomass production as affected by grass species (SPP) and three levels of

soil phosphorus (P) in a 16-week glasshouse experiment. .................................. 107

Table 17. Analysis of variance table (PROC GLM) for the full model testing the

difference in biomass production for grass species (SPP) colonized with arbuscular

mycorrhizal fungi (AMF) and non-AMF-inoculated plants. Dependent variables

shoot difference (SDIFF), root difference (RDIFF), and total difference (TDIFF)

were produced by subtracting biomass of non-AMF plants from AMF plants in a

paired-pot glasshouse experiment examining the effects of AMF source and P on

growth of prairie SPP grown in sterilized mine soil. .......................................... 108

Table 18. Interaction of arbuscular mycorrhizal fungi (AMF) from the Claridon

tallgrass prairie remnant (CL) and the Wilds reclaimed calcareous mine soil (WL)

in Ohio and soil phosphorus concentration (P). Values represent the difference

between AMF-colonized and non-AMF-colonized grasses1 in a 16-week

glasshouse experiment measuring shoot (SDIFF), root (RDIFF), and total biomass

difference (TDIFF) .............................................................................................. 109

Table 19. Interaction of grass species (SPP) and soil phosphorous concentration (P)

on the difference in biomass between grasses colonized with arbuscular

mycorrhizal fungi (AMF) and non-AMF colonized grasses in a 16-week

glasshouse experiment measuring shoot (SDIFF), root (RDIFF), and total biomass

difference (TDIFF). ............................................................................................. 110

xiv

Table 20. Soil properties of reclaimed mine soil and arbuscular mycorrhizal fungi

(AMF) pot-culture soil used in a 10-week glasshouse study examining the

affects of soil compaction and AMF on the growth of three grass species. ....... 130

Table 21. Arbuscular mycorrhizal fungi (AMF) root colonization in three grasses

growing in mine soil during a 10-week glasshouse study. ................................. 131

Table 22. Interaction between grass species and arbuscular mycorrhizal fungi (AMF)

on growth parameters in sterilized mine soil. Sources of AMF are the Wilds

(WL), a 30-year old reclaimed surface mine area near Cumberland, OH, and the

Claridon (CL) tallgrass prairie remnant near Marion, OH, in a 10-week

glasshouse experiment. ....................................................................................... 132

Table 23. Canopy cover estimates of tall fescue (FEAR), Kentucky bluegrass (POPR),

and bird‘s-foot trefoil (LOCO) three years after the 2005 plantings and two years

following the 2006 planting, following re-establishment in native grass plots. . 166

xv

List of Figures

Figure 1. Surface water depth (a) and soil moisture content in the top 15 cm (b) of

reclaimed mine soil at the Wilds study site from April 22 through May 28, 2008.

Reference line in (b) with long dash lines (— — —) indicates soil water potential

at 0.0 MPa, or soil completely saturated. Short dash reference line (– – –) in (b)

indicates soil water potential at -0.3 MPa, or soil at field capacity. Pearson

correlation coefficient between water depth and moisture percent r = -0.69

(P<0.0001). ............................................................................................................ 42

Figure 2. Soil profile of contoured reclaimed calcareous mine soil in southeast Ohio.

Note the color change from dark brown surface layer (approximately 20 cm thick)

to the grayish sub-soil layer below. The surface layer is composed of stockpiled

topsoil or substitute that was spread over the gray-cast overburden during

reclamation. ........................................................................................................... 43

Figure 3. Comparison of per-plant aboveground biomass (grams per plant) for big

bluestem, little bluestem, and Indiangrass in the first and second year of

establishment on reclaimed coal mine land in southeast Ohio. Differences

between species at each harvest were determined with Fisher‘s protected LSD

(α=0.05), and represented with lower case letters (a, b, c) for YR1; differences

between species for YR2 were not significant (ns). Species are represented by

xvi

lines and symbols as follows: big bluestem —●—; little bluestem ···○···;

Indiangrass – –▼– –. .............................................................................................. 86

Figure 4. Affect of arbuscular mycorrhizal fungi (AMF) on number of leaves

produced by prairie grasses at three concentrations of soil phosphorus (P) level

during a 16-week glasshouse experiment. Solid lines and circles (●—●) represent

AMF colonized; dotted lines and open circles (○····○) represent non-AMF

colonized plants. P-values comparing AMF and non-AMF leaf counts are

signified as follows: = 0.05<P<0.001, = 0.001<P<0.0001, = P<0.0001,

and ns = P>0.05. .................................................................................................. 111

Figure 5. Relationships between grass species and soil compaction in sterile mine soil

inoculated with arbuscular mycorrhizal fungi in a 10-week glasshouse study.

Species are tall fescue (—●—), big bluestem (– –■– –), and eastern gamagrass

(····▲····). Differences among compaction levels, for each species, are compared

with Fisher‘s protected LSD (α=0.05) and are shown by the following letters for

each species: tall fescue (a,b,c), big bluestem (j,k,l), eastern gamagrass (x,y,z),

and ns=non-significant. ....................................................................................... 133

Figure 6. Muskingum Mine near Cumberland, OH. Compacted reclaimed land in

foreground persists in non-native forage complex while non-compacted reclaimed

land in background supports forest vegetation. Photo taken early spring,

2004. .................................................................................................................... 161

Figure 7. Contoured compacted mine soil with non-native forage complex and

runoff collecting in low-lying swales. Photo taken early spring, 2004. ............. 162

xvii

Figure 8. Eastern deciduous forests in southeast Ohio near Cumberland, OH. Photo

taken Autumn, 2005. ........................................................................................... 163

Figure 9. Second-year growth of grasses in field plots on reclaimed mine soil at the

Wilds. Plot with dense cover in foreground is tall dropseed, while lighter green

grass with wider leaves in middle of plot area is eastern gamagrass. Non-native

forage vegetation persists around the plot area. Photo taken June 5, 2006. ....... 164

1

Chapter 1: Introduction

Humans have profoundly and negatively affected most natural ecosystems on

the planet, and the expanding demand for natural resources will perpetuate this trend.

In North America alone, diverse prairies and forests have been completely lost, either

replaced by a few species of economic interest, or completely abandoned. The effects

of lost or degraded native ecosystems are seen across the planet as watershed loss

leads to flooding, carbon stored for centuries is released and contributes to global

warming, and species become threatened or extinct as their habitat disappears. To

reverse these trends, humans must become actively involved in restoring damaged

ecosystems to their original integrity. The major problem, however, is that when

ecosystem degradation reaches a specific threshold, complete restoration is unlikely

because original native species may be unable to reestablish, even with attempts to

remediate the disturbed habitat (Ries, 1993). After the point-of-no-return, restoration

efforts must focus on building a new functional ecosystem that will provide positive

ecosystem services and be obtainable (Bradshaw, 1988).

In southeast Ohio, thousands of hectares have been surfaced-mined for coal on

land that once supported vast hardwood forest communities (Kaster and Vimmerstedt,

1996). Restoration of the original forest has not been successful at any level,

2

primarily due to extreme soil compaction that occurred during post-mining

reclamation operations. In place of the original forest species, non-native pasture

grasses and legumes were planted and continue to persist as a low-diversity forage

complex (See Appendix A). While this complex does protect the thin layer of topsoil

from eroding, it does not have the functional qualities of either a native prairie or

forest. It is habitat to few animals because of its limited structural diversity, species

planted are relatively shallow rooted and provide little soil development or carbon

storing potential, and low species diversity leaves unfilled niches available for

invasion by non-native invasive plants.

Since restoration of the original forest ecosystem has failed, an alternative

native ecosystem model must be explored. Establishment of a diverse prairie system

would be more desirable than the current forage complex and may help remediate

existing adverse soil conditions. Native prairie ecosystems are diverse, containing

multiple species of grasses and other herbaceous plants, and are noted for their ability

to increase soil quality and support complex faunal communities (Kline, 1997).

Furthermore, diverse prairie ecosystems are excellent sources of biofuel energy and

are more efficient than high-input corn or soybean systems (Tilman, 2006). However,

since reclaimed mined land is not a naturally occurring habitat in Ohio, there is no

historical record of native species suited to this disturbed environment. Therefore,

establishing a diverse ―native‖ prairie ecosystem on reclaimed mine land must be an

inventive process (rather than restorative), in which we select and evaluate species that

can fill niches in this new habitat (Gross, 2003).

3

Since grasses are the major component of all prairie systems, they are critical

for prairie establishment on reclaimed mine land. Grasses were responsible for

building the deep, rich soils of the Midwest ―corn-belt‖ and supporting the vast herds

of bison on the American Great Plains. However, grasses that establish on reclaimed

mine land must be able to tolerate adverse soil conditions. Reclaimed mine soils are

highly compacted, which limits nutrient and water uptake and reduces root exploration

(Soil Survey Staff, 1996). Low nutrient status, especially of phosphorus, and loss or

inadequacy of mycorrhizae may also be barriers for establishment. Mycorrhizae are

important symbiotic associations between plant roots and soil fungi where the fungi

transfer phosphorus from the soil to the plants in exchange for plant-synthesized

carbohydrates. Many prairie grasses depend on this symbiosis for survival but it is

unknown if the reclaimed mine land have adequate mycorrhizae to meet this need.

The overall goal of this research is to study North American prairie grass

establishment on calcareous reclaimed mine land in southeast Ohio and explore

mycorrhizae and soil compaction as limitations to establishment in an effort to

develop a diverse prairie ecosystem on this disturbed landscape. To achieve this goal,

separate studies examine the following specific objectives: 1) suitability of North

American prairie grasses on reclaimed mine land, 2) adequacy of mine soil

mycorrhizae for native grass growth, and 3) the relationship between soil compaction

and mycorrhizae on grass establishment and growth.

4

Chapter 2: Prairie Grass Establishment on Calcareous Reclaimed Mine Soil.

Abstract

Reclamation of surface mined land in southeast Ohio has created habitats

incapable of supporting native deciduous forest communities. Compaction of

overburden and topsoil restricts plant root and water movement. Limestone in the

overburden produces calcareous soil that is low in available phosphorus. Competition

from non-native forage species inhibits native plant establishment. Replacing the

existing low-diversity forage complex with diverse native prairie communities would

increase ecosystem function; however, a native model for reclaimed mine land prairies

does not exist. Fifteen grasses native to grasslands across North America were planted

on reclaimed mine land in southeast Ohio. Seeds were incorporated into bare soil

during May 2005 and 2006. Population density was assessed at 30 days after planting

and in October of 2005 through 2007. Eastern gamagrass, tall dropseed, buffalograss,

and slender wheatgrass were most successful in the first year of establishment. Big

bluestem, little bluestem, and Indiangrass had the least emergence, although

Indiangrass was more successful in 2006. Eastern gamagrass, tall dropseed, and big

bluestem maintained their population density throughout the study period while

Western wheatgrass had the greatest increase. Big bluestem and little bluestem

5

established poorly, but maintained their population density through the study period.

Blue grama and sideoats grama declined following the first year. Arid-land grasses

Snake River wheatgrass and Sandberg‘s bluegrass failed to establish. Species most

successful were able to tolerate compacted calcareous mine soil that was seasonally

saturated during spring and sometimes droughty during summer. Species that

established well in this research represented species that either diminish the effect of

soil compaction by rooting through the compacted layers, or, that avoid the

compaction by rooting shallow and tolerating any water stress that occurs.

Keywords: Calcareous reclaimed mine soil, native grasses, disturbed land, ecosystem

restoration, grass establishment, southeast Ohio.

Introduction

The loss of ecological memory and resilience occurs as habitats are severely

disturbed and are unable to recover naturally (Nyström and Folke, 2001; Schaefer,

2009). This is often the scenario with human-manipulated terrestrial systems and can

occur when sources of reproductive material are lost or replaced by invasive species,

or when physical changes in the habitat keep native species from reestablishing. As a

result, newly developing communities will likely have different ecological trajectories

and resulting physiognomy, and may not be able to reach the level of functioning as

the original ecosystem (Seaman, 2007; Dodds, et al. 2008). Consequently, the

challenge for restoration ecologists and land managers is to accurately assess the level

6

of disturbance and the potential of attainable outcomes. Serious problems may arise if

rehabilitation plans fail to consider the degree of disturbance in attempting to restore

or establish communities that have little chance of surviving.

Reclamation and restoration of surface mined land is a challenge due to the

degree of disturbance imposed on the land. Surface mining, either open-pit or

mountain-top removal, is one of the most complete forms of human-caused habitat

alteration and degradation (Fischer and Fischer, 2006). Although surface mining in

the U.S. is regulated by state and federal laws which require reclamation of mined

areas, it is impossible to completely restore the original soil profile, soil structure, seed

bank, micro- and macro-biota, and landscape form that existed before mining. In

addition, emergent properties associated with the undisturbed landscape (community

interactions, food webs, nutrient cycling, aesthetics, etc.) cannot be restored simply by

putting pieces (topsoil, topography, plants, etc.) back together.

Furthermore, changes to soil pH and structure are inevitable. Layers of

geologic material are mixed throughout the reconstructed landscape and can

significantly change the character of the profile (Indorante, et al, 1981). In areas

where limestone (calcium carbonate) is a component of the geologic material above

the coal seams (overburden), calcareous soil conditions are created. Calcareous soils

are high in pH and low in available phosphorus (P) as excess calcium binds with P to

produce insoluble precipitates (Brady, 1984; McBride, 1994). Consequently, plants

must depend on adaptive mechanisms or microorganisms to survive in these low-

nutrient conditions (Kothari, et al., 1990; Sanchez, 2007). Furthermore, creating a

7

calcareous soil where a non-calcareous soil existed before may filter out the native

flora making it difficult for native communities to establish.

Coal has been mined since the early 1800‘s in the unglaciated hill country of

the Allegheny Plateau region, but it wasn‘t until the development of large-capacity

earth-moving equipment in the 1900‘s that surface mining had the potential to impact

large areas of the landscape. The native vegetation in this region is primarily eastern

deciduous forest communities (Braun, 1950; Goebel and Hix, 1996). Prior to 1972,

surface mining consisted of complete vegetation removal followed by excavation of

all overburden, including topsoil. The geologic profile consisted of limestone,

claystone, and sandstone, as well as one or more coal seams (Brant, 1964). During

mining the overburden was pushed aside into previously mined pits (spoil piles) to

gain access to the coal, but reclamation and revegetation was left to the discretion of

the mining company.

In 1972, Ohio passed the Ohio Surface Mining and Reclamation Law that

required mined land to be contoured to approximate the original landscape and

specified that topsoil, or a suitable substitute be replaced. In 1977, the U.S.

government enacted the Surface Mining Control and Reclamation Act (SMCRA,

1977), which also required contouring and topsoil replacement. The federal law went

one step further in requiring that vegetation of the original community be

reestablished; however, a loophole in the law stated that introduced species could be

planted ―to achieve the approved postmining land use plan.‖

8

These laws had both positive and negative implications for restoration.

Contouring the mine spoil produced gently rolling slopes less prone to slippage than

steep slopes on spoil piles; however, contouring also required the use of heavy

equipment, which caused a high degree of compaction. Compaction reduces pore

space causing reduced water holding capacity and restricted root growth. For many

plants, soil bulk density greater than 1.5 g cm-3

in fine textured soils is considered very

restrictive for growth (Kozlowski, 1999). Secondly, replacement of topsoil meant that

the top layer of the reconstructed landscape would have silts and clays potentially

higher in nutrient capacity and biological activity than deeper layers of the

overburden; but, silts and clays are also more susceptible to soil compaction, and

quality of stockpiled topsoil can diminish with inadequate storage (Rives, et al., 1980;

Abdul-Kareem and McRae, 1984; Rokich, et al., 2000). Finally, the ―approved

postmining land use plan‖ specified forage grasses and forbs for agricultural use,

rather than native deciduous forest. These forage species were able to establish well

within the 5-year bond window given the mining companies, but were non-native and

competitive and may have excluded native species from establishing.

The end result is nearly 80,000 hectares of land in Ohio stuck in a permanent,

low-diversity, non-native forage complex (Kaster and Vimmerstedt, 1996) with no

ecological blueprint for recovery. The overriding factors of compaction, competitive

grass cover, and high pH may be a permanent barrier for establishment of deciduous

forest vegetation (Burger et al., 2002). Furthermore, the woody invasive autumn olive

(Elaeagnus umbellata Thunb.) has been able to colonize highly compacted reclaimed

9

mine land (Ingold, 2002), while establishment of native forest species has not occurred

even where seed sources are available.

As an alternative to the existing forage vegetation, diverse prairie communities

may be useful in restoring ecological function to these areas. Diverse prairie

communities are more stable than low-diversity grasslands and are associated with

greater soil development (Jastrow, 1987; Tilman and Downing, 1994). Reclaimed

mine land is an artificially constructed ecosystem; consequently, there are no historical

records or native reference areas to help with restoration planning. The objective of

this research was to examine the establishment and growth of grass species native to

different habitats across North America with the goal of increasing diversity and

function on the compacted calcareous reclaimed mined land. The overall hypothesis is

that by assessing which species are most successful at establishing on the reclaimed

mine soil, generalizations can be made about the nature of the mine land habitat and

its ability to support plants with a range of adaptive traits. For example, if species that

establish well are predominately from drier regions, then it is likely that the physical

conditions of the mine soil are more arid than would normally be expressed by the

mesic climate of the region.

Methods

Study Area

This study was initiated in March of 2005 on reclaimed surface mined land

near Cumberland, Ohio. The area is part of the Allegheny Plateau of southeast Ohio,

10

which extends westward from the Allegheny Mountains and is a subdivision of the

Appalachian Mountain Range. The landscape is unglaciated rolling hills with

deciduous forest communities dominated by Quercus, Acer, Fagus, and Carya species

(Braun, 1950). Much of the land had been cleared for agricultural purposes beginning

in the 1800‘s; therefore, the existing forests are in various stages of regeneration

following decline of agricultural use in the 1900‘s.

The study site is located on land that had been mined in the early 1980‘s, and

was once part of the Muskingum Mine, then owned and mined by Central Ohio Coal

Company, a subsidiary of American Electric Power Company, Columbus, OH. All

overburden was removed above the coal seams during mining and was piled in

previously mined areas. As required by law, the mine spoil was then contoured to

approximate the original ―rolling hill‖ landscape and then covered with stockpiled

topsoil or substitute material approximately 20 cm deep. A mix of non-native forage

grasses and legumes were seeded to establish a vegetation ground cover.

It is unknown exactly what species were planted during reclamation, but a

survey of vegetation at the study site in 2005 included tall fescue, Kentucky bluegrass,

and bird‘s-foot trefoil (Table 1). Other forage species found nearby included smooth

brome (Bromus inermis Leyss.) and meadow foxtail (Alopecurus pratensis L.). Non-

forage species included Canada thistle (Cirsium arvense (L.) Scop.), an introduced

weedy species, common milkweed (Asclepias syriaca L.), a native perennial forb, and

autumn olive, an invasive woody species.

11

Plot establishment

The area of interest is located on a 37-km-2

portion of the Muskingum Mine

donated by the Central Ohio Coal Company in 1986 to The International Center for

the Preservation of Wild Animals, Inc. (the Wilds). To this day, this land is managed

by the Wilds for conservation and educational purposes. Plots were set up in April

2005 near the base of a southwest facing slope on a reclaimed spoil ridge. The overall

plot area measured 32x24 m and consisted of four blocks each containing 16 2x6-m

plots. The experimental design was a randomized complete-block with four

replications for each planting. Each plot was further subdivided into three 2-m2

subplots to facilitate plantings in both 2005 and 2006. In April 2005, dead material

and debris was raked off the plot area to expose growing grass. All existing

vegetation was then killed using glyphosate herbicide at a rate of 4.6 kg active

ingredient (a.i.) ha-1

. Four check plots containing the existing forage complex were

left un-sprayed. In accordance with label recommendations, NH4SO4 was added to the

spray mix at a rate of 20 g l-1

to aid movement of the herbicide into the plant. On May

7, 2005, the dying plant material was mowed with a rotary lawn mower. Thatch and

surface soil were loosened with a Bluebird® power dethatcher, and the remaining

plant debris was raked off the plot area by hand. To loosen the soil for planting, plots

were raked with a heavy steel-tined rake to a depth of approximately 2-3 cm.

12

Species selection and seeding

Grass species selected for this study are native to North American grassland

ecosystems (Hitchcock, 1971; Stubbendieck et al., 1986; Klips, 2004); however, each

species has its own preferred habitat where it is more likely to be a dominant or

prominent component of the community (Table 1). Selection areas include the

tallgrass prairies of the Midwest and eastern North America, mixed and short-grass

prairies of the Great Plains region, and species from the intermountain-west shrub

steppe. These grassland ecosystems represent a moisture gradient across North

America from mesic to arid as follows: tallgrass>mixed-grass>short-grass>shrub

steppe.

Warm-season tallgrass prairie species were eastern gamagrass (Tripsacum

dactyloides L.), big bluestem (Andropogon gerardii Vitman), tall dropseed

(Sporobolus asper (Michx.) Kunth), Indiangrass (Sorghastrum nutans (L.) Nash), and

sideoats grama (Bouteloua curtipendula (Michx.) A. Gray). Cool-season grasses

primarily associated with tallgrass prairie were Virginia wildrye (Elymus virginicus

L.) and slender wheatgrass (Elymus trachycaulus (Link) Gould ex Shinners); however,

slender wheatgrass is also found abundantly in the northern Great Plains. Warm-

season grasses primarily associated with mixed- and short-grass prairies were little

bluestem (Schizachyrium scoparium (Michx.) Nash), blue grama (Bouteloua gracilis

(Willd. Ex Kunth) Lag. Ex Griffiths), and buffalograss (Buchloe dactyloides (Nutt.)

Engelm). Cool-season grasses in the mixed- and short-grass regions were western

wheatgrass (Pascopyrum smithii (Rydb.) A. Löve), green needlegrass (Stipa viridula

13

Trin.), and prairie Junegrass (Koeleria macrantha (Ledeb.) J.A. Schultes); however,

prairie Junegrass also occurs frequently in western grasslands. Cool-season grasses

primarily associated with the shrub-steppe of the intermountain west include and

Sandberg bluegrass (Poa secunda J. Presl) and Snake River wheatgrass (Elymus

wawawaiensis J. Carlson & Barkworth). (Table 1).

Seeds for all species except eastern gamagrass were hand-broadcasted into

their respective plots, 1 m2 at a time. After broadcasting, seeds were incorporated into

the loose soil to a depth of 2 cm with a steel-tined rake. Eastern gamagrass seeds were

individually planted to a depth of 1-2 cm. Buffalograss burs that remained on the

surface were pushed into the soil by hand. For both eastern gamagrass and

buffalograss, the seed/burs were difficult to cover by raking alone because of their

large size. Once seeds were sown, the seedbed was firmed up using foot pressure.

Planting in both years occurred during May when the soil was dry enough to loosen

with a rake and get seeds incorporated.

All seeds, except tall dropseed, eastern gamagrass, Sandberg bluegrass, and

Snake River wheatgrass were purchased from Western Native Seeds, Coaldale, CO

USA. Tall dropseed was purchased from Oak Prairie Farm, Pardeeville, WI USA.

Eastern gamagrass was purchased from the Gamagrass Seed Company, Falls City, NE

USA. Sandberg bluegrass and Snake River wheatgrass seed were purchased from

Grassland West Co., Clarkston, WA USA.

14

Plot Management

Following seeding, supplemental irrigation of 15 mm was applied on June 27

and August 9, 2005 as soil surface was drying and cracking. In 2006, supplemental

irrigation of 7 and 5 mm was applied on May 25 and 29, respectively, to moisten soil

that had lightly crusted following a recent rain. Irrigation was only applied as an aid

and would likely not have been needed in 2005 if precipitation would have followed

average patterns, or in either year if seeds had been drilled into the soil instead of

broadcasted and raked (Diboll, 1997).

On August 23, 2005, a dense stand of Canada thistle growing in all plots was

sprayed with mix of 2,4-D amine, mecoprop, dicamba, and sulfetrazone herbicides at

rates of 0.8 kg, 0.28 kg, 0.08 kg acid equivalent (a.e.), and 0.03kg a.i. ha-1

,

respectively. This was a delayed application so that newly seeded grasses had

sufficient growth, at least 5 leaves, to escape injury from the herbicides. Application

was made with a CO2-pressurized backpack sprayer and a 4-nozzle spray wand

applying 140 l spray mix ha-1

.

Prior to the 2006 plantings, plots were sprayed with glyphosate herbicide on

May 2 at a concentration of 20 g a.i. l-1

. On June 5, 2006, Canada thistle growing in

the 2005-planted plots was sprayed with clopyralid herbicide at a concentration of 7.2

g a.e. l-1

. In both applications, leaves were thoroughly covered with spray solution

using a Solo® backpack sprayer and hand-held single-nozzle wand.

On June 26 2007, Canada thistle and other dicotyledonous weeds were sprayed

in all plots with tryclopyr herbicide at an application rate of 1.0 kg a.e. ha-1

. At the

15

same time, witchgrass (Panicum capillare L.) seedlings in the cool-season grass plots

were sprayed with fenoxyprop herbicide at a rate of 0.22 kg a.i. ha-1

. Witchgrass

seedlings in the warm-season grass plots were sprayed with sulfometuron methyl

herbicide at a rate of 0.02 kg a.i. ha-1

. Fenoxyprop is effective at selective controlling

annual warm-season grass seedlings growing in cool-season perennial grass stands,

while sulfometuron methyl at low rates is effective at controlling annual warm-season

grasses in perennial warm-season grass stands without causing injury to the perennial

grasses at the applied rates (Peters et al., 1989).

To eliminate the effect of canopy shading, the plot area was mowed after each

year‘s growth when warm-season grasses were dormant, leaving a stubble height of

10-15 cm.

Population measurements

Population densities were counted at 30 days after planting (30DAP) to assess

early establishment following the 2005 and 2006 plantings, and then during October of

2005, 2006, and 2007 to measure change in populations over time. Counts were made

within a 1-m2 frame placed in the center of each 2x2-m subplot to avoid border effect.

In plots with dense and even populations, counting was limited to 0.25-m2 and

adjusted up to a 1-m2 basis. For buffalograss and western wheatgrass, all rooted nodes

on stolons or stems arising from rhizomes were counted as individual plants. This

reduced confusion during counting but still assessed the status of each population.

16

Ground water monitoring wells

Proximity of ground water to the soil surface and associated soil moisture

content was measured in 2008 between April 1 and May 28 at weekly intervals.

Ground water was measured in 18 monitoring wells systematically placed across the

plot area. Wells measured 90 cm deep by 5.5 cm in diameter and were dug with a

hand-powered bucket auger. A 3.5-cm diameter PVC pipe measuring 100 cm long

was placed in the center of each well. Holes 5 mm in diameter were drilled the length

of each pipe at 5-cm increments to facilitate water flow. Pipes were covered with

plastic-mesh window screen to keep rocks and other material out of the pipe. The pipe

was surrounded by course silica sand to keep soil from plugging the 5-mm holes. At

the soil surface, mortar mix was poured around the pipe to seal the top of the well.

Ground water depth was measured as the distance from the soil surface to the water

level in the pipe. In addition to water depth, soil moisture content was measured at the

same time ground water was measured. Two 2x15-cm soil cores were extracted near

each monitoring well and placed in a sealable plastic bag. Sample bags were taken to

the lab where soil was weighed wet, placed in a 55 ºC drying oven for a minimum of

48 h, then re-weighed dry. Gravimetric soil moisture content was determined by

dividing the soil dry weight (g) by the soil wet weight (g).

Soil compaction and soil moisture characteristics

Soil compaction (g dry soil cm-3

) was determined at 0-5, 5-10, 10-15, and 20-

25 cm depths of the mine soil profile. Two 5-cm diameter soil cores, 0-15 cm and 20-

17

35 cm deep were collected from each of the four plots left in the original forage

complex with a slide-hammer, removable-sleeve soil coring device. Each 15-cm core

was divided into three 5-cm lengths using individual 5-cm sleeves placed in the coring

device. The 20-25 cm depth was collected from the second core taken in the same

hole. The individual 5-cm long cores were weighed wet, dried at 55 ºC for 96 h, then

re-weighed. Weight of the sleeve was subtracted from both the wet and dry weights.

Bulk density was calculated on a wet and dry basis by dividing the soil weight (g) by

the sleeve volume (cm3).

The relationship between soil water content and soil water potential (soil

moisture characteristics) was determined for the top 15 cm of mine soil. A single

5x15-cm core was collected from each of the four check plots, as described above, and

then divided into three 5-cm lengths. Water potentials of 0.0, -0.003, and -0.01

megapascals (MPa) were determined with a gravity tension table. Water potentials of

-0.03, -0.1 and -0.3 MPa were determined using a ceramic pressure plate and negative

vacuum pressure (Richards, 1947).

Statistical analysis

All population data were transformed by adding 0.5 to the raw count and then

calculating the quarter-root of the sum. This transformation was used to improve

conformance with assumptions inherent in analysis of variance (ANOVA) statistical

tests. Transformed data were analyzed using PROC GLM in SAS/STAT® software

(SAS, 2007). Protected Fisher‘s LSD test was used to separate means within

18

comparisons of interest and differences were accepted only if the P-value calculated

by the GLM test was at or less than 0.05 (Milliken and Johnson, 1984). Means were

back-transformed for presentation in tables.

Results and Discussion

Precipitation and soil moisture

The mean precipitation in this region is approximately 1000 mm yr-1

, which is

within the 762-1270 mm yr-1

range given for the eastern deciduous forest biome

(Raven and Berg, 2004). On average, spring and summer months have greater

precipitation than autumn and winter (Table 2). During 2005, precipitation at the

Wilds study site was above average in April and plots were too wet for planting until

May. Precipitation was below average in June and July, but above average in August

(Table 2). In 2006, precipitation in March and April was near average, but ground

water was again at or near the soil surface into April. Precipitation in June and July

was above average but below average in August. As a result, seedlings in 2005

experienced drier conditions into the summer than those in 2006. In contrast,

precipitation in 2007 was well below average in April, May, and June. Drought stress

on established grasses was evident early in the growing season and persisted through

fall. The above average precipitation in July and August was too little and too late to

overcome earlier shortages.

Soil saturation during late winter and spring is a common occurrence at this

site (personal observation). Measurements in 2008 showed the ground water table

19

near or at the soil surface well into May (Figure 1a) with a trend toward a high

perched water table during spring, especially following rain events. Furthermore,

when the water table was high, soil moisture remained above field capacity (Figure

1b). The perched water table and subsequent saturated soil is likely related to the

compacted soil profile and is problematic for early seeding operations and for species

that are intolerant of anaerobic soil conditions.

Soil analysis

Federal surface mining law requires that surface-mined land be contoured

during reclamation to approximate the original landscape, and that a layer of topsoil,

or the best available subsoil, be spread over the contoured overburden (SMCRA,

1977). An exposed profile at the Wilds shows a distinct difference that occurs at

about 20 cm below the soil surface where the brown and brownish-red colors of the

topsoil layer gives way to grayish hues of the contoured overburden (Figure 2). Soil

tests confirmed the calcareous nature of the reconstructed soil profile as pH was above

7.0 and calcium (Ca) levels high at all depths sampled (Table 3). Reclaimed soil in

this region is classified as the Morristown series, which consists of a 20-cm surface

layer of dark brown silty clay loam above light gray silty clay loam overburden that is

alkaline and contains 10 to 40 percent coarse fragments (Soil Survey Staff, 1996). In

contrast, the adjacent non-mined forest soil had slightly acidic pH and much lower Ca

levels. Both sites were non-saline, but, the reclaimed mine soil had higher levels of

Ca and potassium (K) than did the forest soil. High Ca reduces available P as calcium

20

phosphate precipitates are formed (Brady, 1984; McBride, 1994). This is evident in

the mine soil as P is low at all depths compared with the forest soil.

Soil pH was consistently basic in each 5-cm layer of the 20-cm replaced

topsoil layer; however, soil organic matter (SOM) and available P were both greater in

the 0-5 cm surface layer (Table 4). This is likely a result of root growth and decay that

occurred since the land was reclaimed and seeded with non-native forage species,

including the sod-forming Kentucky bluegrass. Furthermore, Ca and K were each

relatively high throughout the 20-cm profile when compared to the non-mined forest

soil (Table 3). Bulk density of the 0-5 cm layer averaged 1.05 Mg m-3

, while depths

below this layer were more compact with bulk densities ranging between 1.5 and 1.8

Mg m-3

(Table 4).

Population dynamics

Emergence of seedlings 30DAP in 2005 and 2006, with the exception of

eastern gamagrass, was consistently low compared with the number of viable seeds

sown (Table 5). However, statistical analysis showed that tall dropseed, Indiangrass,

buffalograss, and slender wheatgrass had 1.8, 1.7, 7.2, and 2.5 times greater

emergence in 2006, respectively, compared with 2005 (Table 5). Big bluestem also

appeared to have greater emergence in 2006; however, in the 2005 30DAP census,

data from only one of the four blocks were available due to a recording error; hence,

no statistical comparison could be made between 2005 and 2006. The 2006 plantings

were on plots that had been fallowed since May 2005 and the seedbeds may have been

21

more suitable for germination of these species. Because of the fallowing, the 2006

plots were also easier to till than plots in 2005. Furthermore, the decaying Kentucky

bluegrass roots had a year to release nutrients and lose any allelopathic affect that may

have been present in 2005 (Bosy and Reader, 1995).

Weather conditions may have also been a factor in the difference in

establishment between 2005 and 2006. Greater precipitation in June of 2006

compared with June of 2005 (Table 2) meant that establishing seedlings were less

likely to experience drought stress when roots were young and encountering the

compacted soil just below the surface layer. Also, mean temperatures, recorded 17 km

NE of the study site, in May and June of 2005 were 13.4 C and 22.2 C, respectively.

Mean temperatures for the same months in 2006 were 14.3 C and 19.1, respectively.

Compared with 2005, the soil warmed up earlier in May 2006, but stayed slightly

cooler in June.

Eastern gamagrass emergence 30DAP in 2006 was 20 plants m-2

, which was

slightly lower than the 24 plants m-2

recorded in the 2005 30DAP census (Table 5);

however, population density was relatively consistent for the two years, especially

when compared with the other species. The consistency of emergence of this species

likely resulted from planting each seed by hand 1-2 cm into the soil, which ensured

that each seed was less affected by fluctuating surface temperature and moisture.

However, high pH and compaction, and low available P were apparently not barriers

for establishment of eastern gamagrass.

22

Emergence for Virginia wildrye, little bluestem, blue grama, and sideoats

grama was similar in 2005 and 2006 (Table 5). Variability among blocks strongly

influenced the analysis, but the exact cause of this variability was undetermined and

may have been related to differences in soil moisture across the plot area.

Species that did not establish in 2005, or were present only at low densities,

were not planted in 2006. These included green needlegrass, prairie Junegrass,

Sandberg‘s bluegrass, and Snake River wheatgrass. A final assessment of these

species was made in May 2006 and a sparse few individuals were found.

Furthermore, no consistency could be seen across the blocks, except for Sandberg‘s

bluegrass, which could not be found in any of the plots (data not shown). These

species may be poorly adapted to the mine soil habitat, general climate, or may

establish better if planted in autumn.

Western wheatgrass planted in 2005 had poor seed viability; therefore,

emergence was near zero (data not shown). Since this species is tolerant of high pH

and moist soil (Table 1), a new seed lot was planted in 2006 and no comparison was

made with the failed 2005 planting. As a precaution, a seed rate twice that used in

2005 was sown in 2006 (Table 5). Establishment density of western wheatgrass in

2006 appeared to be consistent with blue grama, sideoats grama, and big bluestem.

When comparing stand density over time, eastern gamagrass, tall dropseed,

and big bluestem had little change in population from 30DAP through the three years

following planting (Table 6). Final density in YR3 of the 2005 planting of eastern

gamagrass, tall dropseed, and big bluestem density was 24, 104, and 21 plants m-2

,

23

with each density was consistent with earlier censuses. This trend was also persistent

in the 2006 planting and suggests that once these species establish, individuals are

tolerant of the mine soil. Furthermore, tall dropseed was observed growing in plots

adjacent to plots where it was originally seeded, suggesting it is capable of spreading

in this habitat. In contrast, blue grama and sideoats grama quickly diminished in

density after the first year. In the 2005 planting, density of sideoats grama declined

from 19 plants m-2

at 30DAP to 0 plants m-2

at YR3 (Table 6). Blue grama declined

from 130 to 1 plants m-2

over the same period. Similar outcomes were also seen in the

2006 planting. These two species are listed as intolerant of anaerobic soil (Table 1),

which likely explains the decline in density.

Virginia wildrye and slender wheatgrass, in general, established relatively well

but then appeared to decline in density by the final census. Greatest density for

Virginia wildrye in the 2005 planting was 87 plants m-2

at the YR2 census, then

declined to 36 plants m-2

in YR3 (Table 6). Slender wheatgrass density in YR2 of the

2005 planting was 163 plants m-2

, and then declined to just 39 plants m-2

in YR3. By

the end of the study, it was difficult to distinguish individual plants when growing in

dense clumps. If several plants were mistakenly counted as one, then this may

partially explain the decline in density. However, these two cool-season grasses were

highly preferred by deer, rabbits, and rodents especially through the first winter after

planting. Fecal material from these animals was commonly found in the plots and

grazing activity was intense throughout the winter when the warm season-grasses were

dormant. It may be that young slender wheatgrass and Virginia wildrye plants are

24

simply more palatable than the surrounding Kentucky bluegrass or tall fescue, and are

more desirable from a wildlife perspective.

Buffalograss and western wheatgrass both produced dense stands.

Buffalograss, which spreads by stolons as well as seed, increased during the first year

of the 2005 planting from 33 seedlings 30DAP to 205 rooted plants at the end of the

first year (Table 6). In 2006, no difference could be detected for buffalograss from the

30DAP census through the second year census as initial germination was greater than

in 2005. Furthermore, both buffalograss and western wheatgrass spread out of the

plots in which they were seeded. Western wheatgrass substantially increased its

presence from 62 plants m-2

in the first year to 1360 plants m-2

by the end of the

second year following planting (Table 6).

Population density expressed as percent of viable seeds sown was used to

compare establishment success among species. At each of the four censuses of the

2005 planting, eastern gamagrass stand density average 60% or greater of viable seeds

initially sown, which was greater than all other grasses (Table 7). In the 2006

planting, species with the greatest emergence at the 30DAP were eastern gamagrass

(52%), tall dropseed (41%), buffalograss (38%), and slender wheatgrass (63%) (Table

7). At the YR1 census, density as percent of seeds sown was greatest for eastern

gamagrass (49%), buffalograss (47%), and slender wheatgrass (29%). At the YR2

census, density of eastern gamagrass was 52% of seeds sown, but overall greatest for

western wheatgrass, which had increased from 4% to 154% of seeds sown between the

YR1 and YR2 censuses (Table 7). This increase was primarily a function of new

25

shoots emerging from rhizomes spreading into open gaps left from the initial seeding.

However, of the species with the greatest percent emergence, only eastern gamagrass

and tall dropseed maintained their absolute population density throughout the study

(Table 6).

In contrast, 30DAP emergence of big bluestem, little bluestem, and Indiangrass

was only 5%, 8%, and 5% of viable seeds sown for the 2005 planting, and 8%, 24%,

and 3% in the 2006 planting (Table 7). In both plantings, 30DAP emergence of

Indiangrass was greater than either big bluestem or little bluestem, and in the 2006

planting, Indiangrass percent emergence (24%) was not different from tall dropseed

(41%), which was one of the most successful grasses in this study (Table 7). Greater

emergence of Indiangrass in the 2006 planting may have been due to greater rainfall

during June and July in 2006 when seedlings were germinating (Table 2). Indiangrass

density in the 2006 planting was 17 and 11% of seed sown at the YR1 and YR2

census, respectively, and was similar to tall dropseed densities of 24 and 27% of seeds

sown at the same censuses, respectively. During the first year of growth, seedlings of

the big bluestem, little bluestem, and Indiangrass exhibited signs of stress, including

blotchy leaves, crowns elevated above the soil surface, dull color, and early

senescence (personal observation). Low establishment success of these tallgrass

prairie species may indicate impaired or limited ability to development tallgrass prairie

on reclaimed calcareous compacted mine soil. In contrast, seedlings of eastern

gamagrass, tall dropseed, and buffalograss showed little signs of stress through their

seedling year (personal observation).

26

The results from this study indicate several characteristics of the reclaimed

mine soil habitat that may affect revegetation of the landscape. First, anaerobic soil

conditions can occur following periods of heavy rainfall, especially in spring, which

may filter out intolerant species. Both blue grama and sideoats grama diminished by

the second year following planting. Blue grama is native to the short-grass and mixed-

grass prairies of the Great Plains where moisture is often deficient for establishment

(Laurenroth et al., 1994). It is not unexpected that the mesic climate of southeast

Ohio may be too wet for this species: however, sideoats grama is native to tallgrass

prairie communities from Kansas through Ohio and is found in most southern and

eastern U.S. states (USDA, 2009). Therefore, the exit of sideoats grama from the

mine soil is likely related to saturated soil and not the overall climate. Problems

associated with saturated soil are lack of O2 around the roots and reduction of metals,

e.g. iron and manganese, which are toxic to many plants at higher concentration (Taiz

and Zeiger, 2002).

The moist climate and the periodically saturated soil may benefit the non-

native tall fescue and Kentucky bluegrass. Both species have been developed as

highly competitive forage species and are well adapted to a wide range of soil and

climate conditions. The success of tall fescue and Kentucky bluegrass on the mine

soil is likely due to a combination of two main factors. First, both species are well

adapted to the habitat, and secondly, the habitat is not suitable for more competitive

vegetation, i.e. the native forest species, that would naturally establish through

succession. Tall fescue is adapted to poorly drained soil (Lauriault et al., 2005;

27

Hannaway et al., 1999), while certain Kentucky bluegrass cultivars are adapted to

periods of flooding (Wang and Jiang, 2007). The saturated conditions of the mine soil

are directly related to soil compaction; however, tall fescue and Kentucky bluegrass

are also able to tolerate the compaction. Tall fescue establishes well by rooting

directly into the compacted matrix (Crews, 1984). Kentucky bluegrass avoids the

compaction by rooting primarily in the top 20 cm of the soil (DaCosta, et al., 2004).

The mine soil was least compacted in the top 5 cm (Table 4) and periods of saturation

were less frequent and of shorter duration than at deeper depths (Figure 1a). Soil

organic matter was also greater in the 0-5-cm depth (Table 4), reflecting long-term

root growth of the Kentucky bluegrass in the surface layer.

Poor success of the tallgrass prairie species, big bluestem, little bluestem, and

Indiangrass reflects the general difficulty for late-successional prairie species to

establish on bare soil (Kleijn, 2003; Skousen and Venable, 2008). Late-successional

species often experience low seedling establishment rates and stands are slow to

develop when planted in early-successional environments. Seedling desiccation,

improper seeding method, weed competition, and low seeding rate are reasons given

for poor establishment (Launchbaugh and Owensby, 1970; Potvin, 1993; Masters,

1997; Abbott and Roundy, 2003). In this study, germination rates were lower for both

big bluestem and Indiangrass in 2005 when the climate was warmer and drier, than in

2006 with greater precipitation and lower average temperature. Little bluestem

establishment was equally low in both years. This suggests that desiccation during

seedling growth limits establishment of these late-successional grasses; thus, it may be

28

unreasonable to expect a high emergence percentage in this habitat. However, big

bluestem density remained consistent through the three years of the study, whereas the

2005 Indiangrass and little bluestem plantings declined by the last census.

In contrast, eastern gamagrass, tall dropseed, buffalograss, and western

wheatgrass were the most successful grasses to establish and maintained stand density

throughout this research. Eastern gamagrass, a perennial relative of maize (Zea mays

L.), is native in the tallgrass prairie regions but also extends into the southern USA,

Mexico, Central America, and into South America (Randolph, 1970; Hitchcock,

1971). This species is often found on stream banks and floodplains where

establishment is likely to follow disturbance as seeds require warm temperatures

associated with bare soil (Anderson, 1985). Warm temperatures in May and June in

southeastern Ohio, coupled with ephemeral wetland-like conditions of the mine soil

may explain the establishment success this species. However, persistence of

established plants is likely due to the ability of eastern gamagrass roots to penetrate

compacted soil layers (Clark et al., 1998; Gilker et al., 2002) where they can access

water during drought periods. Furthermore, it is also apparent that eastern gamagrass

tolerates the calcareous nature of the reclaimed mine soil. Ability to withstand

saturated calcareous soil and to root into the compacted layers below the soil surface

represents mechanisms for adaptation to the mine soil habitat.

The success of tall dropseed would not be predicted given the listing in the

USDA-NRCS Plants Database as intolerant to high soil pH and anaerobic soil

conditions (USDA-NRCS, 2009). Tall dropseed is a component of tallgrass prairie

29

communities and produces small seeds that accumulate in the soil surface, where they

can germinate following disturbance (Mitchell et al., 1996; Pérez, 1998). Methods of

plot preparation and seeding in this study likely favored tall dropseed as the small

seeds were able to nestle into soil depressions and were not affected by litter layers or

existing canopies of other species. However, the persistence of this species through

the duration of the study is remarkable as populations did not decline in the plots.

Furthermore, at the end of the study, tall dropseed had begun to spread into plots

adjacent to where it was originally planted (personal observation), indicating

adaptation to the habitat. It is not known how deep this species roots into compacted

soil; however, a closely related species, sand dropseed (Sporobolus cryptandrus

(Torr.) A. Gray) was found more effective at extracting water from 0-30 cm than

Broom snakeweed (Gutierrezia sarothrae Shinners), a shrubby competitor effective at

extracting water from 30-60 cm depths (Wan et al., 1993). This suggests that tall

dropseed may limit its root growth to the surface layers, but is very effective at

extracting water in this zone.

Buffalograss, which spreads by stolons, and western wheatgrass, which

spreads by rhizomes, were able to establish by spreading into gaps where seeds

initially did not germinate. The success of these two species represents mechanisms

that are similar to those of Kentucky bluegrass. All three species tolerate low

precipitation, high pH, and anaerobic soil (Table 1); however, they can also utilize

abundant precipitation or irrigation. The reported maximum rooting depth of

buffalograss (0.9 m) is slightly greater than that reported for Kentucky bluegrass (0.6

30

m) but buffalograss is able to extract greater amounts of water from the profile

(Stewart et al., 2004). Western wheatgrass is capable of rooting to 1.2 m (Frank and

Bauer, 1991). Soil compaction of the mine soil likely limits rooting depth of these

three species to shallower layers; however, their persistence is likely due to their

ability to tolerate water stress that naturally occurs at shallow depths in a compacted

profile during periods of drought.

Replacing the non-native forage complex with a diverse plant community

will require a diverse set of mechanisms to overcome constraints imposed by the

compacted calcareous mine soil. Species that established well in this research

represented species that either diminish the effect of soil compaction by rooting

through the compacted layers, or, that avoid the compaction by rooting shallow and

tolerating any water stress that occurs. Species also must tolerate periodic flooding

and the resulting anaerobic soil conditions. However, late-successional grasses like

big bluestem may also persist, even though early establishment is low, because they

are effective at tolerating stress, including low levels of nutrients such as P (Grime,

1977). A community of grasses that include only the species that established in this

research would increase both structural diversity and species richness over the existing

forage complex. However, a greater number of species that tolerate the compaction

and periodic flooding of the calcareous mine soil would further increase diversity in

this habitat.

Furthermore, a diverse prairie ecosystem that contains ecological resilience

and memory would be more able to respond to disturbance and changes in the

31

environment and still maintain functional quality and provide greater ecosystem

services. Greater soil development and carbon sequestration, improved animal

diversity and habitat, and a preserve for plant species that have been diminished or

eliminated from their original ranges due to human-centered land use, are benefits that

could be realized by prairie establishment on compacted mine soil in the Appalachian

coal mining region.

31

Table 1. List of grasses planted in this research and a description of habitat characteristics where each species is native.1

Common name Scientific name

PPT

range

pH

range

Anaerobic

tolerance Habitat; Distribution

---------------------------------------------------------Native prairie grasses--------------------------------------------------------------------

Eastern gamagrass Tripsacum dactyloides L. 406 –

1524

5.1 –

7.5

Medium Facultative wetland species; Gulf

coast into Midwest and

southeastern U.S.

Virginia wildrye Elymus virginicus L. 914 –

1397

5.0 –

7.0

Medium Moist ground, low woods, and along

streams; Great Plains, Midwest,

and eastern U.S.

Big bluestem Andropogon gerardii Vitman 305 –

1397

6.0 –

7.5

Medium Prairies, open woods, and dry

slopes, but may occur near

wetlands; Great Plains, Midwest,

and eastern U.S.

Tall dropseed1 Sporobolus asper (Michx.)

Kunth

406 –

1270

5.5 –

7.0

None Prairies and sandy meadows; Great

Plains, Midwest and northeast

U.S.

Little bluestem Schizachyrium scoparium

(Michx.) Nash

305 –

1143

4.8 –

8.0

None Prairies, open woods, and dry


wetlands; Great Plains through

eastern U.S.

Indiangrass Sorghastrum nutans (L.) Nash 279 –

1143

5.0 –

8.4

Low Prairies, open woods, and dry


wetlands; Great Plains through

eastern U.S.

Continued

32

32

Table 1. Continued.

Common name Scientific name

PPT

range

pH

range

Anaerobic

tolerance Habitat; Distribution1

Western wheatgrass Pascopyrum smithii (Rydb.)

A. Löve

203 –

914

4.5 –

9.0

Medium Moist, usually alkaline soil; western

U.S., Great Plains, and Midwest

Sideoats grama Bouteloua curtipendula

(Michx.) A. Gray

152 –

635

5.5 –

8.5

None Plains, prairies, and rocky hills;

Great Plains and Midwest

Buffalograss Buchloe dactyloides (Nutt.)

Engelm

178 –

813

6.5 –

8.0

High Dry plains; Great Plains

Slender wheatgrass Elymus trachycaulus (Link)

Gould ex Shinners

203 –

635

5.6 –

9.0

Low Moist and well drained soil along

rivers and in meadows; western

U.S., Great Plains, Midwest, and

northeastern U.S.

Green needlegrass Stipa viridula Trin. 381 –

610

6.6 –

8.4

None Plains and dry slopes; Upper

Midwest through Great Plains and

into southwest U.S.

Blue grama Bouteloua gracilis (Willd. Ex

Kunth) Lag. Ex Griffiths

203 –

559

6.6 –

8.4

None Plains; Great Plains and Midwest

Prairie Junegrass Koeleria macrantha (Ledeb.)

J.A. Schultes

356 –

508

6.0 –

8.0

None Prairies, open woods, and sandy

soil; western U.S., Great Plains,

and Midwest

Sandberg‘s bluegrass Poa secunda J. Presl 203 –

406

6.0 –

8.0

None Plains, dry woods, rocky slopes;

western U.S. into Great Plains

Snake River

wheatgrass

Elymus wawawaiensis J.

Carlson & Barkworth

na na NA Dry slopes; Pacific Northwest.

Continued

33

33

Table 1. Continued.

Common name Scientific name PPT

range

pH

range

Anaerobic

tolerance Habitat; Distribution1

---------------------------------------------------Existing non-native vegetation---------------------------------------------------------------

Tall fescue Festuca arundinaceae Schreb. 762 –

1651

5.0 –

9.0

Low Roadsides, meadows, and pastures;

introduced from Europe, widely

cultivated in northern U.S. states.

Kentucky bluegrass Poa pratensis L. 610 –

1651

5.0 –

8.4

Low Open woods, meadows, except arid

areas; introduced from Europe,

widely distributed at all latitudes

below alpine.

Bird‘s-foot trefoil Lotus corniculatus L. 711 –

1651

5.0 –

7.7

Medium Pastures; introduced from Eurasia,

widespread in the U.S.

1Precipitation, pH ranges, and anaerobic tolerance from USDA, NRCS. 2009. The PLANTS Database (http://plants.usda.gov,

22 April 2009). National Plant Data Center, Baton Rouge, LA 70874-4490 USA. Habitat description from Hitchcock,

A.S. 1971. Manual of the grasses of the United States. Ed. A. Chase. Dover Public., New York, and, Stubbendieck, J.,

S.L. Hatch, and K.J. Hirsch. 1986. North American range plants. 3rd

Edition. Univ. of Nebraska Press, Lincoln, NE.

34

http://plants.usda.gov/

35

Table 2. Precipitation data recorded 17 km northwest of the Wilds study site at

the Zanesville, Ohio municipal airport.

Month 2005 2006 2007 1988-2007

------------------------------------(mm)---------------------------------

January 213 72 99 80

February 46 30 34 56

March 67 70 133 78

April 104 75 61 88

May 100 80 32 105

June 85 102 56 95

July 64 114 117 96

August 195 38 150 93

September 76 133 71 87

October 80 148 80 68

November 94 35 61 71

December 23 43 85 62

Annual 1149 942 980 979

36

Table 3. Soil analysis of reclaimed calcareous mine soil and adjacent non-mined

forest soil collected from a former surface coal mine in southeast Ohio.

Parameter ---Reclaimed mine soil--- ---Non-mined forest soil---

Sampling depth (cm) 0-20 20-51 51-81 0-20 20-51 51-81

pH 7.3 7.4 7.6 6.4 5.8 5.7

EC (dS m-1

) 0.1 0.5 0.4 0.11 0.10 0.14

P (mg Kg-1

) 12.1 2.6 2.3 16.0 13.0 8.0

Ca (mg Kg-1

) 3768 4162 6859 952 437 2403

K (mg Kg-1

) 161 112 107 89 70 43

SOM (%) 4.9 2.3 2.0 4.5 2.4 1.1

NO3-N (mg Kg-1

) 8.5 5.0 5.8 4.0 2.8 5.1

Nitrogen (g Kg-1

) 1.7 0.8 0.7 1.8 0.7 0.4

Carbon (g Kg-1

) 30.0 16.0 19.0 23.9 8.2 3.0

C:N 17.9 19.0 27.4 13.0 11.6 7.1

1 Samples were collected June 2004 with a 6.4 cm diameter probe from four

reclaimed mine sites and an adjacent non-mined forest soil site.

37

Table 4. Properties of soil collected from the top 20 cm, in 5-cm increments,

from reclaimed coal mined study site in southeast Ohio. Samples collected

randomly from the Wilds study site on October 2007.

Depth1 pH SOM P K Ca Bulk density

2

(cm)

(%) ---------(mg Kg-1

)--------- (Mg m-3

)

0-5 7.5 5.8 19 233 4091 1.05

5-10 7.6 1.6 4 225 5608 1.51

10-15 7.9 1.0 2 168 5869 1.65

15-20 7.7 0.7 3 183 6630 1.743

1 Six samples were collected from four plots containing pre-existing vegetation

(See Table 3.) and were bulked together. Individual samples measured

2x20 cm and were partitioned into their corresponding depth.

2 Samples for bulk density were collected with a slide-hammer soil probe

containing a 3.2x15 cm sleeve.

3 15-20 cm depth was not available; value is from 20-25 cm depth.

38

Table 5. Population density 30 days after planting (30DAP) in 2005 and 2006

in relation to number of viable seeds sown.

Population density 30DAP

2

Species1

Viable seeds

sown 2005 2006 P-value

(seeds m-2

) ---(count m-2

)--

Eastern gamagrass 39 24 20 0.0337

Tall dropseed 428 101 177 0.0112

Big bluestem 474 25 39 ---

Indiangrass 453 62 108 0.0146

Little bluestem 566 28 18 0.3434

Sideoats grama 432 19 30 0.7090

Blue grama 924 130 62 0.2230

Buffalograss3 633 33 238 0.0120

Virginia wildrye 434 49 77 0.5529

Slender wheatgrass 424 103 259 0.0430

Western wheatgrass4 882 --- 62 ---

Green needlegrass 432 T --- ---

Prairie Junegrass 878 T --- ---

Sandberg‘s bluegrass 815 0 --- ---

Snake River wheatgrass 431 T --- ---

1 Species with trace (T) or zero densities in 2005 were not planted in 2006.

2 P-value compares population density between the 2005 and 2006 planting for

each species. No data available for western wheatgrass in 2005 due to a

poor seed lot. Green needlegrass, prairie Junegrass, Sandberg‘s bluegrass

and Snake River wheatgrass were not did not establish in 2005 and were

not replanted in 2006.

3 Seeding rate refers to whole burs.

39

Table 6. Changes in population density measured 30 days after planting (30DAP), in October of the first year following

planting (YR1), in October of the second and third year after sowing (YR2 and YR3), respectively.

2005 planting2 2006 planting

3

Species1

Viable seeds

sown 30DAP YR1 YR2 YR3 30DAP YR1 YR2

(seeds m

-2) -----------------(plants m

-2)----------------- ----------(plants m

-2)----------

Eastern gamagrass 39 24 a 30 a 33 a 23 a 20 x 19 x 20 x

Tall dropseed 428 101 a 111 a 149 a 104 a 177 x 101 x 114 x

Big bluestem 474 25 a 19 a 15 a 21 a 39 x 29 x 29 x

Indiangrass 453 28 a 35 a 15 b 16 b 108 x 77 xy 49 y

Little bluestem 566 28 ab 40 a 12 b 10 b 18 x 18 x 18 x

Sideoats grams 432 19 b 51 a 1 c 0 c 30 x 34 x 6 y

Blue grama 924 130 a 210 a 7 b 1 b 62 x 25 x 0 y

Continued

39

40

Table 6. Continued.

2005 planting2 2006 planting

3

Species1

Viable seeds

sown 30DAP YR1 YR2 YR3 30DAP YR1 YR2

(seeds m

-2) -----------------(plants m

-2)----------------- ----------(plants m

-2)----------

Buffalograss 633 33 b 205 a 237 a 132 ab 238 x 296 x 160 x

Virginia wildrye 434 49 bc 75 ab 87 a 36 c 77 x 23 x 16 x

Slender wheatgrass 424 103 a 163 a 132 a 39 b 259 x 68 y 90 y

Western wheatgrass4 882 --- --- --- --- 62 y 33 z 1360 x

1 Buffalograss and western wheatgrass counts include plants arising from seed as well as clonal shoots; all other counts

refer only to plants arising from seed.

2 Counts in each row for each the 2005 planting date, are compared with a protected Fisher‘s LSD (α=0.05) and are

different only if followed by a different letter (a, b, or c).

3

Counts in each row for each the 2006 planting date, are compared with a protected Fisher‘s LSD (α=0.05) and are

different only if followed by a different letter (x, y, or z).

4 No data for western wheatgrass in 2005 due to a poor seed lot.

40

41

Table 7. Species comparison of stand density as a percent of viable seeds sown, at each census for the 2005 and

2006 plantings.1

2005 planting 2006 planting

Species 30DAP YR1 YR2 YR3 30DAP YR1 YR2

-------- (% of viable seeds sown )-------- -------(% of viable seeds sown)------

Eastern gamagrass 62 a 65 a 85 a 60 a 52 a 49 a 52 b

Tall dropseed 24 b 26 bc 35 b 24 b 41 ab 24 b 27 c

Big bluestem 5 d 4 e 3 de 4 cd 8 de 6 d 6 ef

Indiangrass 8 c 8 de 3 d 4 d 24 bc 17 bc 11 cd

Little bluestem 5 d 7 de 2 def 2 d 3 e 3 d 3 f

Sideoats grama 4 d 12 cde <1 f <1 e 9 de 10 cd 2 f

Blue grama 14 c 24 bc 1 ef <1 e 7 e 3 d <1 g

Buffalograss 6 d 38 b 38 b 22 b 38 abc 47 a 26 c

Virginia wildrye 11 c 18 bcd 20 c 8 c 20 cd 5 d 4 f

Slender wheatgrass 24 b 40 b 31 bc 9 c 63 a 29 ab 21 cd

Western wheatgrass2 --- --- --- --- 7 e 4 d 154 a

1 At each census, percent emergences followed by the same letter are determined not different with a protected

Fisher‘s LSD (α=0.05). 2 No data for western wheatgrass in 2005 due to a poor seed lot.

41

42

(a)

Sample Date

04/22 04/29 05/05 05/13 05/21 05/28

So

il M

ois

ture

Per

cen

t

0

5

10

15

20

25

30

35

0.0 MPa

-0.3 MPa

Su

rfac

e W

ater

Dep

th (

cm)

0

10

20

30

40

50

60

70

(b)

Figure 1. Surface water depth (a) and soil moisture content in the top 15 cm (b) of

reclaimed mine soil at the Wilds from April 22 through May 28, 2008. Reference

line in (b) with long dash lines (— — —) indicates soil water potential at 0.0 MPa,

or soil completely saturated. Short dash reference line (– – –) in (b) indicates soil

water potential at -0.3 MPa, or soil at field capacity. Pearson correlation

coefficient between water depth and moisture percent is r = -0.69 (P<0.0001).

43

Figure 2. Soil profile of contoured reclaimed calcareous mine soil in southeast Ohio.

Note the color change from dark brown surface layer (approximately 20 cm thick)

to the grayish sub-soil layer below. The surface layer is composed of stockpiled

topsoil or substitute that was spread over the gray-cast overburden during

reclamation.

44

Chapter 3: Productivity of Native Prairie Grasses on Reclaimed Calcareous Mine Soil.

Abstract

Reclamation of surface coal mined land in the eastern U.S. has produced

habitats incapable of supporting deciduous hardwood forests native to the region.

Compacted calcareous soil limits restoration and reduces overall ecosystem function

and diversity. Furthermore, competitive non-native forage species planted during

reclamation persist as a low-diversity forage complex. As an alternative, a native

prairie community may increase diversity, ecosystem function, and productivity.

Fifteen grasses native to North American were planted on bare soil in monoculture

plots on reclaimed surface mine land in southeast Ohio. Plantings were in 2005 and

repeated in 2006. Plots were visually assessed for canopy cover and clipped for

aboveground biomass production in October of the first, second, and third year

following planting. Cover and biomass were consistently high for eastern gamagrass,

tall dropseed, and buffalograss, which had produced dense stands. These species

produced less biomass than the existing forage vegetation only in the first year of

growth. In contrast, tallgrass prairie dominants big bluestem, little bluestem, and

Indiangrass had low cover and biomass production. Sideoats and blue grama were

45

productive in the first year but diminished by year two. Western wheatgrass

increased substantially in cover and biomass by the second year from spreading

rhizomes. Slender wheatgrass and Virginia wildrye were productive but results were

mixed in the second planting. Biomass on a per-plant basis was greatest for eastern

gamagrass and lowest for western wheatgrass. Results show that native prairie grasses

can be productive on calcareous reclaimed mine soil.

Keywords: Native grass productivity, disturbed land restoration, surface mined land,

calcareous soil, soil compaction, prairie establishment.

Introduction

Primary productivity is the basis for ecosystem function and is critical in

restoring core processes to degraded landscapes. Carbon sequestration, nutrient

cycling, and community diversity all depend on the ability of producers to transform

inorganic carbon and energy into plant biomass (Martin et al., 2005; Kivimäki et al,

2008; Wu et al., 2008). Reestablishing native communities is a preferred means of

restoring ecosystem function on disturbed land; however, habitat conditions on

degraded sites may limit productivity and may inhibit native plants from establishing.

Surface mined land in the eastern U.S. is permanently impaired by reclamation

that produced adverse soil conditions, primarily soil compaction and high pH, which

prohibit reestablishment of the native deciduous forest communities (Burger et al.,

2002). Soil compaction caused by heavy equipment used to spread and contour spoil

46

and stockpiled topsoil during reclamation is a major factor limiting forest

regeneration. Bulk densities ranging from 1.5 to 1.6 g cm-3

can occur in the top 5 cm

of recently reclaimed land and can range between 1.6 and 1.8 g cm-3

at lower depths

(Shrestha et al., 2009). Bulk densities of 1.5 g cm-3

or greater can seriously reduce

root growth, mineral and water uptake, lead to anaerobic soil conditions, and reduce

plant establishment and productivity (Kozlowski, 1999). Furthermore, in areas with

limestone in the substrata, calcareous soil conditions occur when limestone is mixed

and spread near the surface. Calcareous soils are alkaline in pH and can be critically

low in available phosphorus (Brady, 1984; McBride, 1994).

These habitat conditions are unfavorable for the majority of deciduous trees

native to the region. A study conducted on reclaimed surface mined land in southeast

Ohio found that only white ash (Fraxinus americana L.) and green ash (Fraxinus

pennsylvanica Marsh.) had established successfully (>90%) 5 years after planting,

whereas mature-community dominants such as red oak (Quercus rubra L.) and

Shumard oak (Quercus shumardii Buckley var. shumardii) established poorly

(<20%), and yellow poplar (Liriodendron tulipifera L.), a regional early successional

hardwood species, essentially failed to establish (0.5%) (Kost et al., 1994).

Reclamation of surface coal mined land in the United States is mandated by

Public Law 95-87 (Surface Mining Control and Reclamation Act of 1977), which

requires establishment of a ―permanent vegetative cover of the same seasonal variety

native to the area of land to be affected‖ (SMCRA, 1977). This implies that

reclamation of surface mined lands in the eastern U.S would include establishment of

47

the native deciduous forest species; however, a loophole in the law allows the use of

non-native species if they fit within an ―approved postmining land use plan‖

(SMCRA, 1977). Furthermore, mining companies are given five years to establish a

permanent ground cover if they are to retrieve bond money paid to cover reclamation

costs in the event of abandonment. It is within this context that mining companies

plant non-native forage grasses and legumes that establish easily within the five-year

bond window as alternatives to native forest vegetation. Forages such as tall fescue

(Festuca arundinaceae Schreb.) and bird‘s-foot trefoil (Lotus corniculatus L.) are

selected because they establish well, produce adequate ground cover for soil

conservation, and fit within a pre-approved agricultural use plan. Restoration of

native forests would require restoration of habitat conditions favorable to the native

species. Reclamation law specifies only that the original topographical contour be

restored and that a layer of topsoil, or suitable substitute, covers the reclaimed land

surface, but the law contains no directive on limiting compaction.

Replacing the low diversity, non-native forage vegetation with a diverse

prairie community may be a reasonable alternative for increasing ecosystem function

on compacted, calcareous reclaimed surface mined land. Evidence suggests that

species diversity is associated with greater ecosystem stability and productivity

(Tilman et al., 2006) and that aboveground net primary productivity increases as

species richness increases (Foster et al., 2007; Flombaum and Sala, 2008).

Furthermore, soils that develop under native prairie vegetation have greater aggregate

stability and organic matter (Jastrow, 1987), which would help alleviate compaction

48

and poor aeration problems. However, the key to establishing prairie communities on

reclaimed mine soil rests in finding native prairie species that establish well, are

productive, and are able to persist over time (Evanylo et al., 2005; Casselman et al.,

2006).

Tallgrass prairie ecosystems are found in the eastern part of the North

American central plains (Sala et al., 1988). These prairies are highly productive,

diverse, and are the most mesic of the central plains grassland biome. Because of their

proximity to eastern U.S. coal mining areas, they may be useful models for assembling

prairie communities on reclaimed mined land; yet, the same habitat barriers to native

forest establishment may also apply to prairie species. Consequently, species from

other grasslands systems should be considered in constructing a mine soil prairie.

Compacted calcareous mine soil habitat is potentially very different from the well-

developed prairie soils that support tallgrass prairie. For example, the mine soil

habitat may be more or less mesic compared with what the general climate would

reflect on naturally developed well drained soil not compacted or chemically

compromised. Historically, climate change and associated wildfire were responsible

for the development of the grassland biome throughout central North America

(Axelrod, 1985). Similarly, habitat changes from surface mining and reclamation

practices are responsible for creating new habitats that appear to favor grassland

communities; however, the question remains as to what type of grassland ecosystem

would restore function, diversity, and increase productivity.

49

The goal of this research is to identify prairie grass species that are best suited

to reclaimed calcareous mine soil in southeast Ohio. Suitability of grasses

representing different prairie ecosystems was assessed by measuring productivity and

cover during the first three years of establishment. Species selected occur naturally in

North American tallgrass prairies, mixed and short-grass prairies, and the shrub-

steppes of the intermountain western U.S. We hypothesized that mesophyllic tallgrass

prairie species will be less successful than those more tolerant of arid environments

because of the moisture limitations imposed by soil compaction.

Methods

Study Area

This study was initiated in 2005 on reclaimed surface mined land near

Cumberland, Ohio. The area is part of the Allegheny Plateau of southeast Ohio, which

extends westward from the Allegheny Mountains as a subdivision of the Appalachian

Mountain Range. The landscape is unglaciated rolling hills with deciduous forest

communities dominated by Quercus, Acer, Fagus, and Carya species (Braun, 1950).

However, much of the land was cleared for agricultural purposes beginning in the

1800s; therefore, the existing forests are in various stages of regeneration following

decline of agricultural use in the 1900s.

The study site is located on land that was mined in the early 1980s originally

part of the Muskingum Mine. The land was owned and mined by Central Ohio Coal

Company, a subsidiary of American Electric Power Company, Columbus, OH, but

50

was donated to The International Center for the Preservation of Wild Animals, Inc.

(the Wilds) for conservation and educational purposes. During mining, all overburden

was removed above the coal seams and was piled in previously mined areas. As

required by SMCRA 1977, the mine spoil was then contoured during reclamation to

approximate the original ―rolling hill‖ landscape and covered with stockpiled topsoil

or substitute material approximately 20 cm deep. Non-native forage grasses and

legumes were seeded to establish a vegetation ground cover.

It is unknown exactly what species were planted during reclamation, but a

survey of vegetation at the study site in 2005 included tall fescue (Festuca

arundinaceae Schreb.), Kentucky bluegrass (Poa pratensis L.), and bird‘s-foot trefoil

(Lotus corniculatus L.). Other forage species found nearby included smooth brome

(Bromus inermis Leyss.) and meadow foxtail (Alopecurus pratensis L.). Non-forage

species included Canada thistle (Cirsium arvense (L.) Scop.), a non-native weedy

species, common milkweed (Asclepias syriaca L.), a native perennial forb, and

autumn olive (Elaeagnus umbellata Thunb.), an invasive woody species.

Plot establishment and management

A plot area was selected in April 2005 near the base of a southwest facing

slope on a reclaimed spoil ridge. The overall plot area measured 32x24 m and

consisted of four blocks each containing 16 2x6-m plots. The experimental design

was a randomized complete-block with four replications for each planting. Each plot

was further subdivided into three 2 m2 subplots to facilitate multiple plantings in both

51

2005 and 2006. In April 2005, the plot area was sprayed to kill existing vegetation

using glyphosate herbicide at a rate of 4.6 kg active ingredient (a.i.) ha-1

. Four plots

containing the existing forage complex were left un-sprayed as controls. On May 7,

2005, dying plant material and debris were removed and the soil surface was loosened

with a Bluebird® power dethatcher. Plots were raked with a heavy steel-tined rake to

a depth of approximately 2-3 cm prior to seeding.

Following initial plot area preparation and planting, herbicides were applied to

selectively kill Canada thistle and witchgrass (Panicum capillare L.) in the planted

grass stands. Plots planted in 2006 were re-sprayed prior to planting with glyphosate

herbicide to kill remaining vegetation. See chapter two for specific details.

Species selection

Grass species selected for this study are native to North American grassland

ecosystems (Hitchcock, 1971; Stubbendieck et al., 1986; Klips, 2004). Warm-season

tallgrass prairie species were eastern gamagrass (Tripsacum dactyloides L.), big

bluestem (Andropogon gerardii Vitman), tall dropseed (Sporobolus asper (Michx.)

Kunth), Indiangrass (Sorghastrum nutans (L.) Nash), and sideoats grama (Bouteloua

curtipendula (Michx.) A. Gray). Cool-season tallgrass prairie grasses seeded were

Virginia wildrye (Elymus virginicus L.) and slender wheatgrass (Elymus trachycaulus

(Link) Gould ex Shinners); however, slender wheatgrass is also found abundantly in

the northern Great Plains. Warm-season grasses primarily associated with mixed- and

short-grass prairies were little bluestem (Schizachyrium scoparium (Michx.) Nash),

52

blue grama (Bouteloua gracilis (Willd. Ex Kunth) Lag. Ex Griffiths), and buffalograss

(Buchloe dactyloides (Nutt.) Engelm). Cool-season grass occurring in the mixed- and

short-grass regions were western wheatgrass (Pascopyrum smithii (Rydb.) A. Löve),

green needlegrass (Stipa viridula Trin.), and prairie Junegrass (Koeleria macrantha

(Ledeb.) J.A. Schultes); however, prairie Junegrass also occurs frequently in western

grasslands. Cool-season grasses primarily occurring in the shrub-steppe of the

intermountain west include and Sandberg bluegrass (Poa secunda J. Presl) and Snake

River wheatgrass (Elymus wawawaiensis J. Carlson & Barkworth).

All seeds, except tall dropseed, eastern gamagrass, Sandberg bluegrass, and

Snake River wheatgrass were purchased from Western Native Seeds, Coaldale, CO

USA. Tall dropseed was purchased from Oak Prairie Farm, Pardeeville, WI USA.

Eastern gamagrass was purchased from the Gamagrass Seed Company, Falls City, NE

USA. Sandberg bluegrass and Snake River wheatgrass seed were purchased from

Grassland West Co., Clarkston, WA USA.

Soil compaction and analysis

Soil compaction (g dry soil cm-3

) was determined at 0-5, 5-10, 10-15, and 20-

25 cm depths of the mine soil profile. Two 5-cm diameter soil cores, 0-15 cm and 20-

35 cm deep were collected from each of the four plots left in the original forage

vegetation with a slide-hammer soil coring device with removable sleeves. Each 15-

cm core was divided into three 5-cm lengths using individual 5-cm sleeves placed in

the coring device. The 20-25 cm depth was collected from a second core taken in the

53

same hole. The individual 5-cm segments were weighed wet, dried at 55 ºC for 96 h,

then re-weighed. Weight of the sleeve was subtracted from both the wet and dry

weights. Bulk density was calculated on a dry-weight basis by dividing the dried soil

weight (g) by the sleeve volume (cm3).

Soil samples for chemical analysis were collected in October 2007 from plots

left in the existing forage vegetation. Samples were collected from 0-5, 5-10, 10-15,

and 15-20 cm depths. Samples were analyzed for pH, soil organic matter (SOM),

nitrate nitrogen (NO3-N), phosphorus (P), potassium (K), calcium (Ca), and

magnesium (Mg) by A & L Analytical Laboratories, Inc., Memphis, TN.

Biomass and canopy cover measurements

Aboveground biomass was sampled in all plots during October of 2005, 2006,

and 2007. Sampling was conducted to minimize the amount of disturbance to the

plots. For the 2005 planting, these censuses represented first year (YR1), second year

(YR2), and third year (YR3) growth, respectively. For the 2006 planting, 2006 and

2007 represented YR1 and YR2 growth, respectively. Clipping was limited to the area

outside the center 1 m2 area, which was reserved for population density counts.

Clipped biomass was put in paper bags and dried at 55 ºC for a minimum of 96 h. For

buffalograss and tall dropseed, which produced dense uniform stands, a 20x50-cm

area was clipped and dry weight biomass was calculated on an area basis. For all

other species, biomass was estimated by clipping individual plants to determine

average dry weight per plant, and then multiplying by the population density. For

54

species other than western wheatgrass, 5 representative plants were clipped from each

plot. Several species had low population density and 5 individuals was the minimum

that could be found outside the center 1 m2 area. For western wheatgrass, 100

individual shoots were clipped. For the existing vegetation, a 1 m2 area in the center

of one of the three sub-plots in each main plot was clipped and all biomass was

bagged together. Clipping height was 1 cm for all species except eastern gamagrass,

which was clipped 5 cm above the ground. All biomass measurements were

calculated on a 1 m2 area basis for analysis and comparison. In addition, per-plant

biomass (g dry weight plant-1

) for each species was calculated by dividing biomass (g

m-2

) by population density (plants m-2

).

During the three years of this study, biomass and cover were measured in

October when most warm-season grasses were dormant, or nearly so; however, eastern

gamagrass and buffalograss still had green leaves and appeared to be actively growing.

The cool-season grasses, slender wheatgrass, Virginia wildrye, and western

wheatgrass, and the existing forage species, were all actively producing new leaves

with the cooler autumn temperatures (personal observation). Sampling at this time

was intended to capture the current year‘s growth; however, cool-season grass

biomass may have included some senescent leaf material from the previous autumn

and winter. Furthermore, turnover of leaves and culms during the growing season

could have resulted in underestimation of biomass. During sampling, attention was

given to collecting all living and senescent biomass from the past year‘s growth for

each plant harvested to minimize this potential error.

55

Canopy coverage was estimated visually within a 1 m2 frame placed in the

center of each subplot and assessed on six coverage classes as follows: class 1 = 0-5%,

class 2 = 5-25%, class 3 = 25-50%, class 4 = 50-75%, class 5 = 75-95%, and class 6 =

95-100% canopy cover (Daubenmire, 1959). For consistency, all estimates were made

by the author. The midpoints of each range were substituted for the class number in

the statistical analysis and presentation.


All data were mathematically transformed to improve conformance with

assumptions inherent in analysis of variance (ANOVA) statistical tests. Biomass data

were transformed using the following equation:

(biomass m-2

+ 0.05)0.25

eq. 1

Cover percents were transformed using the following equation:

(arcsine(cover percent / 100))0.5

eq. 2

Per-plant biomass was transformed using the following equation:

(biomass plant-1

)0.25

eq. 3

Transformed data were analyzed using PROC GLM in SAS/STAT® software (SAS,

2007). Protected Fisher‘s LSD test was used to separate means within comparisons of

interest, differences were accepted only if the P-value calculated by the GLM test was

0.05, or less (Milliken and Johnson, 1984). All transformed means were back

transformed for presentation.

56


Yearly fluctuations in precipitation likely had an influence on production and

growth. May and June of 2007 were considerably drier than in 2005 or 2006 (Table

2). In 2006, near-normal precipitation was recorded from March through July;

whereas, in 2007, precipitation was below normal from April through June causing

drought stress to be visible on many of the grasses, including the existing Kentucky

bluegrass and tall fescue.

Soil properties most conducive for plant growth were found in the top 5 cm of

the mine soil profile. In this layer, SOM measured 5.8%, P was 19 mg Kg-1

, and bulk

density averaged 1.05 Mg m-3

(Table 8). Below 5 cm, P was low, ranging between 2

and 4 mg Kg-1

, and soil was more compact with bulk density up to 1.74 Mg m-3

at 20

cm. Furthermore, SOM was only 1.6% at the 5-10 cm and 0.7% at the 15-20 cm

depth. At all depths, pH was basic, ranging from 7.5 to 7.9, reflecting the high

concentration of Ca from limestone deposits in the overburden.

Canopy cover

Canopy cover is a measure of how well a species (or group of species)

occupies space, at least in a two-dimensional plane. For comparison, canopy cover of

the existing forage vegetation in control plots was assessed as one entity and measured

98% cover in all plots all years (Table 9). Individually, Kentucky bluegrass coverage

was assessed at 98% in all plots, whereas tall fescue and bird‘s-foot trefoil cover

averaged 38 and 19%, respectively (data not shown). Canopy cover is important from

57

a soil conservation standpoint; however, Kentucky bluegrass is a vigorous competitor

that produces a thick sod of densely growing rhizomes, which inhibits seedling

establishment of other species (Curtis and Partch, 1948; Sampson and Moser, 1982;

Jackson, 1999) and likely plays a role in maintaining low plant diversity on reclaimed

mine lands. Successful establishment of a diverse community would require

disruption of the existing vegetation so that new species could establish.

In this study, the existing vegetation was removed with herbicides prior to

planting. Species with the quickest development and most consistent maintenance of

canopy cover were buffalograss, tall dropseed, and eastern gamagrass. Canopy cover

for these grasses was estimated between 80 and 90% in all censuses except for the

2006 spring planting of eastern gamagrass, which was only 59% (Table 9). This

demonstrates an ability to compete quickly for space, independent of yearly

fluctuations in weather. Slender wheatgrass and Virginia wildrye planted in 2005 also

covered the soil quickly, averaging 80 and 89% cover in YR1, respectively.

Establishment of these grasses was lower in the 2006 planting (See Chapter 2), which

resulted in lower canopy cover values of 7 and 44%, respectively, in the YR1 census

(Table 9). However, slender wheatgrass cover increased to 84% by the YR2 census of

the 2006 planting. Virginia wildrye was severely grazed by rodents during the 2006

growing season, which may explain the 7% canopy cover measured in YR1 of this

planting.

Big bluestem, little bluestem, and Indiangrass had the lowest canopy cover in

YR1 of the 2005 planting, averaging 20, 20, and 37%, respectively (Table 9). By

58

YR3, big bluestem averaged 74% and was not different from either slender wheatgrass

or Virginia wildrye. Better establishment of Indiangrass in the 2006 planting resulted

in 68 and 74% cover in the YR1 and YR2 censuses, respectively, which was not

different from the existing forage vegetation. Little bluestem cover ranged from 15 to

37% in all censuses, which reflected low establishment density (See Chapter 2).

Western wheatgrass cover was 15% in YR1, but 84% in YR2 as it was able to spread

and fill open gaps left from YR1. Western wheatgrass is tolerant of high soil pH and

apparently not inhibited by periodically saturated soil. In spite of the low cover in

YR1, western wheatgrass seemed well adapted to the mine soil habitat.

The decline in cover for sideoats grama and blue grama reflected their decline

in plant density through the second and third year following planting (See Chapter 2).

Sideoats grama went from 74% cover in YR1 of the 2005 planting to only 2% by YR2

and 0% by YR3 (Table 9). This trend was consistent in both plantings and also

apparent for blue grama as it had only 2% cover in YR3 of the 2005 planting, and 1%

cover in YR2 of the 2006 planting. The quick and complete loss of cover and

presence of these species resulted in bare soil, which was open for reinvasion by the

non-native forage species, or invasion by early successional species represented in the

seedbank or nearby extant populations (Appendix B, Table 23).

In addition to canopy cover, eastern gamagrass had greater vertical structure

than all other species in the study. Leaf canopy height for eastern gamagrass, 3 years

post-planting, measured 90 cm (data not shown). Leaf canopy height for eastern

gamagrass was three times higher than Kentucky bluegrass (30 cm), and more than

59

twice the height of tall fescue (40 cm). All other species, except tall dropseed, had

leaf canopy heights less than 40 cm. From a restoration standpoint, eastern gamagrass

would provide greater structural diversity on reclaimed mine soil. Greater

aboveground structure should also correlate with greater belowground growth and

structure (Fitter, 1986; Tilman and Wedin, 1991; Dwire et al., 2004).

Biomass

Aboveground biomass produced by the existing forage vegetation, which

consisted primarily of Kentucky bluegrass and tall fescue, yielded 421, 440, and 314 g

m-2

in 2005, 2006, and 2007, respectively (Table 10). For comparison, mean biomass

yield of tall fescue monocultures grown in 2005 and 2006 southern Ohio forage

research trials was 1175 and 1324 g m-2

, respectively (Sulc et al., 2006). These yields

were produced on silt-loam soil that was fertilized with N, and represented multiple

clippings per year. On the reclaimed mine soil, the existing non-native forage

vegetation produced roughly one third of that produced by the pure stands of tall

fescue grown on productive agricultural land, and was more similar in yield to two

sites of Nebraska tallgrass prairie where biomass of 400 and 470 g m-2

were recorded

(Mitchell et al., 1996). Furthermore, tall fescue growing on a reclaimed mine soil in

Virginia yielded 458 g m-2

six years after planting, but then yielded only 197 g m-2

on

the seventh year with different climate conditions (Evanylo et al., 2005). At three

southeast Ohio reclaimed mine sites, aboveground biomass 5 years after planting with

forage grass/legume mixes, averaged 329, 288, and 337 g m-2

(Shrestha et al., 2009).

60

These reports suggest that productivity of the reclaimed mine site in this study

was comparable with other reclaimed mine soils in the region, and even with some

tallgrass prairies, but soil compaction, low fertility, and species composition may have

limited potential productivity. Alleviating compaction on reclaimed mine soil would

require deep ripping of the profile with heavy equipment (Croton and Ainsworth,

2007) and could be costly if applied to large areas of land. Increasing fertility is costly

but less difficult than modifying bulk density throughout the profile; however, simply

fertilizing the existing forage vegetation would likely increase biomass at the expense

of diversity (Tilman, 1993; Baer, et al., 2003). If biomass production is the ultimate

goal, i.e. biofuel production, then increasing fertility to benefit highly competitive

species with superior growth potential would be an option, as is the case with corn-

based ethanol production or forage production. However, long-term sustainability

would be compromised if input costs outweigh marginal benefits. Changing

community composition would be more desirable from an ecological perspective as

ecosystems with greater diversity tend to optimize stability and productivity (Tilman

et al., 2006).

In this research, biomass production was higher for the existing forage

vegetation (421 and 440 g m-2

), compared with the other grasses, only in the first year

of production of both the 2005 and 2006 plantings (Table 10). This was anticipated as

new plantings of perennial grasses are normally less productive than established

stands. However, by YR2, biomass production from tall dropseed, buffalograss,

eastern gamagrass, and western wheatgrass were each similar to the existing

61

vegetation. For the 2005 planting, YR2 production of tall dropseed, eastern

gamagrass, and buffalograss averaged 626, 571, and 277 g m-2

, respectively (Table

10). Western wheatgrass had produced 212 g m-2

by YR2 of the 2006 planting, and

was also similar to the existing forage vegetation. Productivity of these native grasses

was similar to the existing non-native forage vegetation in the second year of

establishment, which suggests good pre-adaptation to the habitat. Furthermore, these

grasses had established dense stands (See Chapter 2) and were tolerating the

compacted soil. By YR3 of the 2005 planting, Virginia wildrye had produced 271 g

m-2

and was also similar to the existing forage vegetation (Table 10).

Production of big bluestem, Indiangrass, and little bluestem was consistently

less than the existing forage vegetation in all censuses (Table 10); however, this was

also a function of lower population density (See Chapter 2). These grasses are

dominant components of mixed and tallgrass prairies throughout the North American

central grasslands, but are considered slower to establish than other grasses, especially

non-native cool-season species (Robocker et al., 1953; Hsu and Nelson, 1986; Tilman

and Wedin, 1991). In YR3 of the 2005 planting, biomass produced by big bluestem,

Indiangrass, and little bluestem was only 95, 87, and 49 g m-2

, whereas the existing

vegetation produced 314 g m-2

. In contrast, sideoats and blue grama could not

maintain productivity over the duration of the study. Yield of sideoats grama declined

from 152 to 1 g m-2

between YR1 and YR3 of the 2005 planting, while blue grama

yield declined from 98 to 0 g m-2

in the same planting (Table 10). At the end of the

62

study in 2007, only a sparse few plants of these two species could be found and were

growing only in drier areas of the plot site (personal observation).

Performance of the cool-season slender wheatgrass and Virginia wildrye was

mixed. Virginia wildrye produced greater biomass (271 g m-2

) than slender

wheatgrass (94 g m-2

) only in YR3 of 2005 planting (Table 10). Virginia wildrye did

not yield well in the 2006 planting, producing only 2 and 11 g m-2

in YR1 and YR2,

respectively, but herbivory from rodents was observed and likely reduced the harvest

yield. Slender wheatgrass biomass in YR2 of the 2005 planting (256 g m-2

) was not

different from the existing forage vegetation, but in YR3 yielded 94 g m-2

and was

30% less than the existing vegetation. This suggests that slender wheatgrass may be

productive early during community development but declines with time. Leaves of

slender wheatgrass developed discoloration during summer (personal observation),

which may have indicated a mineral deficiency, such as phosphorus given the

calcareous nature of the mine soil. Virginia wildrye did not show signs of stress at

anytime during the study; however, it is apparently preferred by some herbivores,

which suggests it may be beneficial from a wildlife perspective.

Comparing these grasses on a per-plant basis gives a different assessment than

could be made from comparisons on an area basis, as above. Per-plant biomass for

Indiangrass (2.9 g plant-1

), big bluestem (1.7 g plant-1

), and little bluestem (1.3 g plant-

1), averaged across all three years and both plantings, was not different from tall

dropseed, which averaged 2.2 g plant-1

(Table 11). This is an important comparison

because tall dropseed yielded as well as the existing forage vegetation on an area basis

63

in both YR2 and YR3. In addition, Indiangrass, big bluestem, and little bluestem did

not decline in per-plant biomass from YR1 through YR3. This suggests that once

these species become established, they are productive on the reclaimed mine soil.

Furthermore, eastern gamagrass, which was more robust than all other grasses, had the

greatest per-plant biomass yielding 11.7 g plant-1

. In contrast, sideoats and blue grama

both declined in per-plant biomass during this research. In YR1, sideoats and blue

grama averaged 3.2 and 0.7 g plant-1

, respectively, but averaged only 0.1 and 0.2 g

plant-1

, respectively, by YR2 (Table 11).

Summary

Productivity of grasses planted in this study reflected variability in moisture

conditions during the growing season on the mine soil habitat. Moisture was

consequential, but at different levels. Soil moisture status ranged from standing water

during late winter and spring to very dry during summer months when precipitation

was limited and evaporation greater. During spring months, and especially following

rainy periods, the soil profile was saturated over much of the plot area to the extent

that the water table was at the soil surface (Chapter 2). Both eastern gamagrass and

buffalograss were productive throughout the study, which suggests moisture

limitations due to soil compaction are not necessarily a barrier to species from either

arid or mesic environments. However, excess moisture is a barrier for species

intolerant of anaerobic conditions, such as blue grama and sideoats grama. Sideoats

64

and blue grama were the only species that declined in both per-plant biomass and total

biomass during this research. Both these grasses are listed as intolerant of anaerobic

soil conditions (USDA, 2009), thus their decline is likely related to their intolerance to

the mine soil habitat. These grasses were productive in their first year, but did not

survive the first winter and spring following planting when the soil was saturated.

In general, with the exception of buffalograss, species that were most

productive on the mine soil were native to moist environments. Eastern gamagrass,

tall dropseed, Virginia wildrye, and western wheatgrass are found in mesic grassland

habitats and were most productive in this research in both aboveground biomass and

canopy cover production. In their second year of establishment, these species, with

the exception of Virginia wildrye, equaled the yearly biomass produced by the existing

non-native forage vegetation. Buffalograss is native to the driest region of the Great

Plains and is very tolerant of drought, but also appears tolerant of periodic saturated

soil and is productive in higher rainfall regions. Tallgrass prairie species big bluestem

and Indiangrass established poorly but were productive on a per plant basis,

suggesting the mine soil may be somewhat limiting for seedlings, but less problematic

for established plants.

This research suggests that mesic environments should be referenced for

assembling a diverse prairie community on the reclaimed mine soil. Tallgrass prairies

in the southeast portion of the central grasslands where precipitation is greatest and

species such as eastern gamagrass are prevalent may be useful models. Also, wetland

prairies may also contain species such as sedges and forbs that would be good

65

candidates in constructing a ―native‖ calcareous mine soil community. This research

demonstrates native prairie grasses can be productive on calcareous compacted mine

soil, but species selection must include adaptation to the mine soil habitat.

66

Table 8. Properties of soil collected from the top 20 cm, in 5 cm increments,

from reclaimed coal mined study site in southeast Ohio. Samples collected

randomly from the Wilds study site on October 2007.

Depth1 pH SOM NO3-N P K Ca Mg

Bulk

density2

(cm)

(%) --------------(mg Kg-1

)-------------- (Mg m-3

)

0-5 7.5 5.8 < 5 19 233 4091 940 1.05

5-10 7.6 1.6 < 5 4 225 5608 995 1.51

10-15 7.9 1.0 < 5 2 168 5869 851 1.65

15-20 7.7 0.7 < 5 3 183 6630 986 1.743

1 Six samples were collected from four plots containing pre-existing vegetation

and were bulked together. Individual samples measured 2x20 cm and were

partitioned into their corresponding depth.

2 Samples for bulk density were collected with a slide-hammer soil probe

containing a 3.2x15 cm sleeve.

3 15-20 cm depth was not available; value is from 20-25 cm depth.

67

Table 9. Canopy cover of grasses, measured the first (YR1), second (YR2), and

third (YR3) years following planting, on reclaimed mine soil in southeast Ohio.

2005 Planting2 2006 Planting

3

Species1 YR1 YR2 YR3 YR1 YR2

----------- (%) ----------- --------- (%) ---------

Existing vegetation4 98 a 98 a 98 a 98 a 98 a

Eastern gamagrass 85 ab 98 a 98 a 59 bcd 95 ab

Tall dropseed 85 ab 98 a 98 a 83 ab 95 ab

Big bluestem 20 c 30 bc 74 bc 11 f 62 bc

Indiangrass 37 c 43 b 56 cd 68 abc 74 abc

Little bluestem 20 c 21 bcd 31 d 15 ef 37 cd

Sideoats grama 74 b 2 d 0 e 70 abc 15 de

Blue grama 80 ab 12 cd 2 e 30 def 1 e

Buffalograss 87 ab 98 a 98 a 98 a 98 a

Virginia wildrye 80 ab 98 a 92 ab 7 f 15 de

Slender wheatgrass 89 ab 92 a 88 ab 44 cde 84 ab

Western wheatgrass --- --- --- 15 ef 88 ab

1 Multiple comparisons between species are made for each census using protected

Fisher‘s LSD (α=0.05). Values in each column followed by the same letter are

not different.

2 YR1 = 2005, YR2 = 2006, and YR3 = 2007.

3 YR1 = 2006, YR2 = 2007.

4 Existing vegetation consisted of Kentucky bluegrass, tall fescue, and bird‘s-foot

trefoil.

68

Table 10. Aboveground biomass of grasses, measured in censuses in October of

the first (YR1), second (YR2), and third (YR3) years following planting, on

reclaimed mine soil in southeast Ohio.

2005 Planting year2 2006 Planting year

3

Species1 YR1 YR2 YR3 YR1 YR2

----------- (g m

-2) ----------- ------- (g m

-2) -------

Existing vegetation4 421 a 440 ab 314 ab 440 a 314 a

Eastern gamagrass 114 b 571 ab 496 a 176 b 271 a

Tall dropseed 119 b 626 a 413 ab 111 b 306 a

Big bluestem 8 d 82 de 95 cd 5 cd 72 cd

Indiangrass 31 cd 150 cd 87 cd 94 b 79 bcd

Little bluestem 13 d 30 ef 49 d 10 cd 52 de

Sideoats grama 152 b 3 g 1 e 111 b 15 e

Blue grama 98 b 9 fg 0 e 23 c 0 f

Buffalograss 156 b 277 abc 197 bc 180 b 223 ab

Virginia wildrye 91 b 61 de 271 ab 2 d 11 ef

Slender wheatgrass 83 bc 256 bc 94 cd 25 c 83 bcd

Western wheatgrass --- --- --- 4 cd 212 abc

1 Comparison between species are made at each census with a protected Fisher‘s

LSD (α=0.05). Values in each column followed by the same letter are not

different.

2 YR1 = 2005, YR2 = 2006, and YR3 = 2007.

3 YR1 = 2006, YR2 = 2007.

4 Existing vegetation consisted of Kentucky bluegrass, tall fescue, and bird‘s-foot

trefoil.

69

Table 11. Per-plant biomass of grasses growing on calcareous reclaimed mine soil

in southeast Ohio measured the year of planting (YR1) and in the second

(YR2) and third (YR3) years following planting.

Biomass per plant

Species YR1 YR2 YR3

Mean

-----------(g plant-1

)1 -----------

(g plant

-1)2

Eastern gamagrass 6.0 b 15.3 a 21.7 a

11.7 a

Indiangrass 1.1 b 4.6 a 5.5 a

2.9 b

Tall dropseed 1.1 b 3.3 a 4.0 a

2.2 bc

Big bluestem 0.3 b 3.8 a 4.6 a

1.7 bc

Little bluestem 0.5 a 2.5 a 1.7 a

1.3 bcd

Buffalograss 0.7 b 1.3 a 1.6 a

1.1 cd

Virginia wildrye 0.4 b 0.7 b 7.4 a

1.0 cd

Slender wheatgrass 0.4 b 1.4 a 2.4 a

1.0 cd

Sideoats grama 3.2 a 0.1 b 0.0 b

0.5 de

Blue grama 0.7a 0.2 a 0.0 a

0.2 e

Western wheatgrass 0.1a 0.2 a na

0.1 e

1 The change in per-plant biomass for each species over the three-year census

periods is compared using protected Fisher‘s LSD (α=0.05).

2 Differences between species for mean per-plant density are compared using

protected Fisher‘s LSD (α=0.05).

70

Chapter 4: Safe-sites Control Warm-season Prairie Grass Establishment on

Calcareous Compacted Mine Soil.

Abstract

Restoring native vegetation on surface mined land can be limited by available

propagules or safe sites where seedlings can establish. Surface mining in the eastern

U.S. has altered the landscape such that native forest species are unable to establish in

compact soil following reclamation. In place of forest species, non-native forage

species were planted and persist as a low-diversity forage complex. Native prairie

communities would impart greater diversity and function, but it is not clear if

increasing seed density would increase establishment success. This research examined

seeding rate on establishment of three dominant North American prairie grasses on

reclaimed calcareous mine soil in southeast Ohio. Big bluestem (Andropogon gerardii

Vitman), Indiangrass (Sorghastrum nutans (L.) Nash), and little bluestem

(Schizachyrium scoparium (Michx.) Nash) are prairie dominants in late-successional

prairie communities but are recognized as slow to establish. Seeds of each species

were sown at 1x and 2x rates onto bare-ground plots in May, 2006. Stand density and

plant biomass were assessed 30 days after planting (30DAP), and in autumn of the

first year (YR1) and second year (YR2) after planting. Density of each species sown

71

at the 2x rate was greater at 30DAP, but not different from the 1x rate at YR1 or YR2.

Indiangrass experienced the greatest decline through YR2. Per-plant biomass was

greatest for Indiangrass in YR1, but no differences between species were found in

YR2. Only big bluestem and little bluestem plants increased in biomass between YR1

and YR2. Results indicate that population density is more related to safe site

availability than seed density, and that mechanisms of establishment differ between

species.

Keywords: Native grass establishment, seeding rate, safe sites, calcareous mine soil,

soil compaction.

Introduction

As Harper (1977) describes, ―the presence or absence and the density of a

seedling population depends not only on the availability of seed but on the frequency

of ‗safe sites‘ that provide the precise conditions required by a particular seed‖ to

germinate and develop into a viable plant. This dichotomy is realized in restoration of

disturbed land where habitat changes and propagule loss have altered the ‗normal‘

response to disturbance. Restoration often requires that species be reintroduced into

disturbed areas because of limited propagule availability (Foster and Tilman, 2003).

However, variability of microsites, i.e. combinations of mineral nutrients, space, light,

moisture, temperature, ground cover, competition, and soil exist across the landscape

and strongly affect seedling survival (Vogel, 1987; McKenna et al., 1991; Oswald and

72

Neuenschwander, 1993; Burke and Grime, 1996; Masters, 1997; Getzin et al., 2008).

Consequently, poor stand establishment can result from either insufficient seed

number or from a limited number of microsites where resources are adequate for

germination and establishment of individual plants.

Reestablishing native vegetation on reclaimed surface mined land can be a

major challenge. Seed reserves in stockpiled topsoil are often inadequate to restore

disturbed land to pre-mined native communities (Iverson and Wali, 1982; Bellairs and

Bell, 1993). Therefore, seeds or live material, e.g. tree seedlings, must be planted

during reclamation to have any chance of reaching restoration goals. However,

reclaimed mine soil can differ structurally and chemically compared with the pre-

mined soil, and may not be capable of supporting native vegetation, i.e, inadequate

safe sites. Mine spoil can range in pH from acid to alkaline and be too low in nutrients

to support plant growth. Reclamation practices using heavy equipment to spread and

contour overburden can cause structural problems, such as soil compaction, which

impedes root growth and water permeability. For example, in the Appalachian coal

mining region of the eastern U.S., reclamation of surface coal mined land since the

enactment of the Surface Mining Control and Reclamation Act of 1977 (SMCRA,

1977) has produced highly compact soils incapable of regenerating the native

deciduous forest vegetation (Burger et al., 2002; Casselman, et al., 2006). In place of

native hardwood trees, these lands have been re-vegetated with highly competitive

non-native forage grasses and legumes that further inhibit native plant seedlings from

establishing (Evanylo et al., 2005; Casselman, et al., 2006). In areas where limestone

73

sediments compose a portion of the overburden, calcareous conditions persist that

maintain alkaline pH and reduce the availability of phosphorus (Brady, 1984;

McBride, 1994). These conditions together alter the successional processes that would

regenerate hardwood forest, and favor a low-diversity forage species complex.

Establishing prairie communities would be an alternative for restoring diversity

and increasing structural and functional qualities to reclaimed mined land. Prairie

species native to the region, include warm-season grasses big bluestem, little

bluestem, and Indiangrass (Hitchcock, 1971; Klips, 2003; Klips, 2004). These species

are dominant components of tallgrass and mixed-grass prairies in the North American

central grasslands (Weaver, 1931); however, establishment is often slow compared

with non-native species such as tall fescue. In a related study (Chapter 2), big

bluestem, Indiangrass, and little bluestem establishment was sparse compared with

other prairie grass sown on reclaimed mine soil.

The objective of this study was to determine if establishment of big bluestem,

Indiangrass, and little bluestem on calcareous reclaimed mine soil is affected more by

seed rate or limited number of safe sites. Poor establishment would leave open gaps

that could be colonized by weedy species, thus slowing establishment of a prairie

community. If increasing seed rate would benefit stand establishment, then fewer

open gaps would be available. However, if establishment is limited by safe sites, then

no amount of seed is going to increase establishment success.

74

Methods

Study Area

The area of interest is part of the Allegheny Plateau of southeast Ohio, which

extends westward from the Allegheny Mountains and is a subdivision of the

Appalachian Mountain Range. The landscape is unglaciated rolling hills with

deciduous forest communities dominated by Quercus, Acer, Fagus, and Carya species

(Braun, 1950). The study site is on reclaimed surface mined land are near

Cumberland, Ohio as part of a larger study examining native grass alternatives to non-

native forages (See Chapter 2). The site is located on land that had been mined in the

early 1980‘s by Central Ohio Coal Company, a subsidiary of American Electric Power

Company, Columbus, OH. During mining, all overburden material above the coal

seams was removed and, as required by reclamation law, then contoured to

approximate the original ―rolling hill‖ landscape during reclamation. The contoured

overburden (mine spoil) was then covered with a layer of stockpiled topsoil or

substitute material approximately 20 cm deep, and seeded with a mix of non-native

forage grasses and legumes to establish a vegetation ground cover.

The reclaimed soil in this region is classified as the Morristown series, a deep

and well drained soil with slow permeability that contains free lime and coarse

fragments with slopes ranging from 1 to 70 percent (Soil Survey staff, 1996). The top

20 cm consists of a dark brown silty clay loam soil with 10 percent coarse fragments

of shale, siltstone, and limy shale spread over the contoured overburden as a topsoil

replacement. Below 20 cm, the profile consists of light gray silty clay loam

75

overburden that is alkaline and contains 10 to 40 percent coarse fragments. Bulk

density measured at the study site averaged from 1.05 g cm-3

at the surface to 1.74 g

cm-3

at 20 cm, while pH measured 7.5 to 7.9 (See Chapter 2). Available phosphorus

measured 19 mg Kg-1

and organic matter 5.8% in the top 5 cm but considerably less at

lower depths. During plot preparation, tillage was kept to a minimum to conserve

nutrients and structure in the surface layer.

Plot establishment

The experimental design was a randomized complete-block with four

replications for each planting. Whole plots measured 2x6 m and were subdivided into

three 2-m2 subplots to facilitate paired-plot comparisons between 1x and 2x seeding

rates. The third subplot was used for a different component of the study. Plots were

initially prepared in April 2005 near the base of a southwest facing slope on a

reclaimed calcareous spoil ridge. Pre-existing vegetation, consisted primarily of

Kentucky bluegrass (Poa pratensis L.), tall fescue (Festuca arundinaceae Schreb.),

and bird‘s-foot trefoil (Lotus corniculatus L.). All vegetation was killed in 2005 using

glyphosate herbicide at a rate of 4.6 kg active ingredient (a.i.) ha-1

. The soil surface

was then lightly tilled and loosened with a Bluebird® power dethatcher. Dead and

dying debris was removed from the plot area and the plots were fallowed through

2005 using glyphosate to control vegetation regrowth.

76

Plot Management

Prior to planting in 2006, plots were re-sprayed in early-May with glyphosate

herbicide to kill vegetation that had re-established through the winter and spring. Two

weeks after spraying, plots were tilled with a steel-tine rake to prepare a loose 2–3 cm

deep seedbed. In late-May 2006, seeds were broadcasted into each plot and

incorporated with a steel-tine rack. Species planted were ‗Bison‘ big bluestem

(Andropogon gerardii Vitman), ‗Tomahawk‘ Indiangrass (Sorghastrum nutans (L.)

Nash), and ‗Itasca‘ little bluestem (Schizachyrium scoparium (Michx.) Nash). Seeds

were purchased from Western Native Seeds, Coaldale, CO USA. Two seeding rates

were applied for each species. Big bluestem was seeded at 474 (1x) and 948 (2x)

viable seeds m-2

, Indiangrass was seeded at 453 (1x) and 906 (2x) seeds m-2

, and little

bluestem was seeded at 566 (1x) and 1132 (2x) seeds m-2

(Table 12). Following

planting, supplemental irrigation of 7 and 5 mm was applied on May 25 and 29, 2006,

respectively, to moisten soil that had lightly crusted following a heavy rain soon after

planting.

In 2007, Canada thistle and other dicotyledonous weeds were selectively

controlled with tryclopyr herbicide on June 26 at an application rate of 1.0 kg acid

equivalent (a.e.) ha-1

. At the same time, sulfometuron methyl herbicide was applied at

a rate of 0.02 kg a.i. ha-1

to kill a flush of witchgrass (Panicum capillare L.) seedlings.

Sulfometuron methyl at low rates is effective at controlling annual warm-season

grasses in perennial warm-season grass stands without causing injury to the perennial

grasses at the applied rates (Peters et al., 1989).

77

Population and biomass measurements

Population densities were counted at 30 days after planting (30DAP) in 2006

to assess early establishment, and then during October of 2006 (YR1) and 2007 (YR2)

to measure change in population density over time. Counts were made within a 1-m2

frame placed in the center of each 2x2-m subplot to avoid border effect. Aboveground

biomass was also sampled during October of 2006 and 2007 by collecting five

individual plants from each subplot, clipped 1 cm from the soil surface in the area

surrounding the center 1 m2 area. Biomass from each plot was bagged and dried in a

55 ºC oven for a minimum of 96 h, and then weighed to determine mean dry weight

per plant (per-plant biomass). The mean per-plant biomass was multiplying by the

population density to determine production on an area basis.


All data were mathematically transformed to improve conformance with

assumptions inherent in analysis of variance (ANOVA) statistical tests. For

population density and stand biomass, data were transformed by taking the square root

of each measurement. Per-plant biomass data were transformed by calculating the

log10 of each value. Transformed data were analyzed using PROC GLM in

SAS/STAT® software (SAS, 2007). Protected Fisher‘s LSD test was used to separate

per-plant biomass means within comparisons of interest and differences were accepted

only if the P-value from the GLM test was 0.05, or less (Milliken and Johnson, 1984).

All transformed means were back transformed for presentation.

78


Overall, establishment rates were low for all three species. Big bluestem,

Indiangrass, and little bluestem had maximum 30DAP emergence of 8%, 23%, and

5% of viable seed sown, respectively (Table 12). However, low germination rates are

not unusual for these grasses. Cornelius (1944) averaged only 6.5%, 17.9%, and 3.3%

establishment rate of viable seeds sown for big bluestem, Indiangrass, and little

bluestem, respectively, during three consecutive years of a tallgrass prairie

revegetation study in Kansas. Seeding rate was not discussed as a factor; however,

Masters (1977) reported an increase in stand frequency in two Nebraska sites when

seeding rate of big bluestem was increased from 220 to 440 viable seed m-2

.

In our study, differences between seeding rates for each species were more

evident at 30DAP than at YR1 or YR2. The 30DAP counts in June 2006 for big

bluestem, Indiangrass, and little bluestem were 1.8, 1.9, and 3.0 times greater at the 2x

than the 1x seeding rates, respectively, but these margins diminished by the YR1

census (Table 12). At the 30DAP census, big bluestem densities averaged 69 and 39

plants m-2

for the 1x and 2x seeding rates, respectively, and the comparison was nearly

significant at α=0.05 (P=0.0664). However, at the YR1 and YR2 censuses, densities

were nearly identical between the two seeding with associated P-values of 0.8020 and

0.7677, respectively. This occurred because of a declining trend in big bluestem

density between the 30DAP and YR1 and YR2 censuses. Density at the 2x rate

declined from 69 to 33 plants m-2

(P=0.0585) whereas, density at the 1x rate declined

from 39 to 29 plants m-2

(P=0.0994).

79

Indiangrass populations responded similarly to big bluestem at the 30DAP

census with germination reflecting the difference between seeding rates. Densities of

109 and 201 plants m-2

were counted at the 1x and 2x rates, respectively, with an

associated P-value of 0.0653 (Table 12). At the YR1 and YR2 censuses, differences

between rates were not found. However, between the 30DAP and YR2 censuses,

Indiangrass declined in density from 201 to 69 plants m-2

at the 2x rate (P=0.0006),

and from 108 to 49 plants m-2

at the 1x rate (P=0.0407). These differences clearly

reflect greater seedling mortality than was seen for big bluestem.

Little bluestem population at the 30DAP census was found to be greater at the

2x rate than the 1x rate (P=0.0285) with densities of 54 and 18 plants m-2

, respectively

(Table 12). But again, differences could not be detected between rates at the YR1 and

YR2 censuses, even though means were 2.5 and 1.6 greater for the 2x rate,

respectively. There was also no detectable change in density from 30DAP to YR2 for

either seeding rate. Stand density at the 2x rate averaged 54 plants m-2

at 30DAP and

29 plants m-2

in YR2. At the 1x rate, 18 plants m-2

were counted at each census.

For each species, difference between the 1x and 2x seeding rates did not

remain consistent throughout the course of the study (Table 12). Densities at the 2x

rate tended to be greater at the 30DAP census, but this was not evident at the YR1 or

YR2 censuses. This interaction would suggest that population density on reclaimed

mine soil is limited by the number of safe sites and not seed density. Given the rocky,

compacted soil, it is likely that the safe sites were related to areas conducive to root

development. The surface 0-5 cm of the mine soil contained greater organic matter

80

and nutrients and was less compact than lower layers. At depths below 5 cm the soil

was compact and may have been a barrier to root growth and access to moisture.

However, rocks in the mine soil profile may have contributed to seedling survival

because roots growing along rock faces can access greater depths and moisture than

roots restricted by impenetrable layers (Nobel, et al., 1992; Haering, et al., 2004).

Grass seedlings that have access to greater soil moisture are more likely to

survive than those exposed to dry conditions (Stubbendieck and McCully, 1972;

Frasier et al., 1984). In this study, seedlings likely experienced dry soil during

summer months between periodic rainfall events. Soil moisture is critical for the

development of secondary roots as moist periods of 3 days or more are required for

their initiation and elongation (Newman and Moser, 1988a). Seedling mortality

occurring between the 30DAP census (June 2006) and the YR1 census (October 2006)

could have been related to moisture stress if seedlings were not able to develop

secondary roots. By October of 2006, surviving seedlings were dormant and some

crowns were observed elevated above the soil surface. Warm-season grasses tend to

elevate their crowns during germination due to elongation of the subcoleoptile

internode (Newman and Moser, 1988b). It is possible that seedlings germinating from

shallow depths elevated their crowns above the surface before secondary roots could

establish, and then died from drought stress.

Stand biomass measurements were similar between seeding rates for all

species at both YR1 and YR2 censuses (Table 13). This is consistent with the

similarity in population density between seeding rates. However, stand biomass for

81

big bluestem and little bluestem increased from YR1 to YR2, whereas Indiangrass

production did not differ between years. From YR1 to YR2 at the 1x rate, big

bluestem biomass increased from 5 to 72 g m-2

(P<0.0001) and little bluestem biomass

increased from 11 to 54 g m-2

(P=0.0062). At the 2x seeding rate, big bluestem

biomass increased 8 to 61 g m-2

(P=0.0099) and little bluestem biomass increased 27

to 85 g m-2

(P=0.0263). Indiangrass seeded at the 1x rate, produced 96 g m-2

in YR1

and 81 g m-2

in YR2 with a comparison P-value of 0.4333. At the 2x rate, biomass

measured 157 and 115 g m-2

at the YR1 and YR2 censuses, respectively, with a

comparison P-value of 0.2506.

However, when biomass was examined on a per-plant basis, each of the three

species differed at the YR1 census (Figure 3). In YR1, Indiangrass plants were largest

averaging 1.8 g plant-1

, followed by little bluestem yielding 0.3 g plant-1

and big

bluestem averaging 0.02 g plant-1

. But in YR2, differences could not be detected

between the three species as Indiangrass, little bluestem, and big bluestem averaged

3.1, 11.5, and 5.5 g plant-1

, respectively. Between YR1 and YR2, the change in per-

plant biomass was significant for big bluestem (P<0.0001) and little bluestem

(P<0.0001), but not for Indiangrass (P=0.2094). This is partly because big bluestem

and little bluestem plants were smaller than Indiangrass plants in YR1; however, it is

unclear why Indiangrass plants did not gain in biomass during YR2.

Greater growth in the first year of establishment may or may not be associated

with likelihood of long-term survival (Huston and Smith, 1987). Individuals of early

successional species typically grow vigorously soon after disturbance and then decline

82

in abundance and productivity during later years as individuals of late-successional

species become dominant. However, all three of the species in this study are

considered late-successional species in the North American central grasslands, but

they may differ in early development. For big bluestem and little bluestem, seedlings

that germinated in the first year acquired little aboveground biomass and only survived

in the most favorable microsites. In the second year, surviving individuals

substantially increased in biomass. However, Indiangrass seedlings were equally

productive during the first and second years of growth; yet, early biomass

accumulation apparently did not prevent seedling mortality.

This study suggests that a limited number of available microsites where

seedlings can germinate and survive through the first year of growth may be a limiting

factor for establishment of these three late-successional prairie species. Increasing

seeding rate does not appear to increase population density. This may be problematic

if the goal is to quickly establish a dense stand that will compete with pioneer and

non-native species, such as Kentucky bluegrass and tall fescue seeded during

reclamation. Competition from the non-native forage species would likely exclude

many bluestem or Indiangrass seedlings. Forage species were initially planted that

established quickly and were pre-adapted to the mine soil conditions, i.e. compaction,

basic pH, low permeability, etc.

Given the results from this study, it is clear that increasing seeding rate alone

would be insufficient to establishing warm-season late-successional prairie grasses. If

late-successional prairie species are to have a chance, strategies must be used to allow

83

the seedlings time to establish without competition from existing or re-establishing

non-native species. Prior to planting, existing vegetation must be removed with

herbicides or tillage. Herbicides, such as atrazine or metolachlor applied prior to

planting (Vogel, 1987; Masters, 1997), or sulfometuron methyl at low rates for

selective grass weed control following planting (Peters et al., 1989) could release the

prairie grasses from competition. Furthermore, a fallow year prior to seeding using

glyphosate to control all vegetation, as was used in this study, is helpful in slowing the

reestablishment of non-native species, which are likely well represented in the seed

bank.

84

Table 12. Effect of seeding rate on stand establishment of three warm season

native grasses on reclaimed calcareous mine spoil.

Species Seeding rate1 30DAP

2 YR1

3 YR2

4 P-value

----------(plants m

-2)---------

Big bluestem 2x 69 34 33 0.0585

1x 39 29 29 0.0994

P-value

0.0664 0.8020 0.7677

Indiangrass 2x 201 116 69 0.0006

1x 108 77 49 0.0407

P-value

0.0653 0.1958 0.2718

Little bluestem 2x 54 45 29 0.1149

1x 18 18 18 0.9874

P-value

0.0285 0.1187 0.1797

1 Big bluestem 1x rate = 474, 2x rate = 948 seeds m

-2; Indiangrass 1x rate = 453,

2x rate = 906 seeds m-2

; little bluestem 1x rate = 566, 2x rate = 1132 seeds m-2

.

2 30DAP census occurred 30 days after planting to assess early emergence.

3 YR1 census occurred in October of the first year following planting.

4 YR2 census occurred in October of the second year following planting.

85

Table 13. Effect of seeding rate on stand biomass of three warm season native

grasses on reclaimed calcareous mine spoil.1

Species Seeding rate YR1 YR2 P-value

--------(g m

-2)--------

Big bluestem 2x 8 61 0.0099

1x 5 72 <0.0001

P-value

0.5588 0.6911

Indiangrass 2x 157 115 0.2506

1x 96 81 0.4333

P-value

0.3312 0.2039

Little bluestem 2x 27 85 0.0263

1x 11 54 0.0062

P-value

0.1084 0.1959

1 See Table 12 for description of seeding rates and censuses.

86

YR1 YR2

Ab

ov

egro

un

d p

er-p

lant

bio

mas

s (g

pla

nt-1

)

0.01

0.1

1

10

100

(YR1-YR2; P=0.2094)

(YR1-YR2; P<0.0001)

(YR1-YR2; P<0.0001)

a

b

c

ns

Figure 3. Comparison of per-plant aboveground biomass (grams per plant) for big

bluestem, little bluestem, and Indiangrass in the first and second year of

establishment on reclaimed coal mine land in southeast Ohio. Differences

between species at each harvest were determined with Fisher‘s protected LSD

(α=0.05), and represented with lower case letters (a, b, c) for YR1; differences

between species for YR2 were not significant (ns). Species are represented by

lines and symbols as follows: big bluestem —●—; little bluestem ···○···;

Indiangrass – –▼– –.

87

Chapter 5: Effectivity of Arbuscular Mycorrhizal Fungi of Reclaimed Calcareous

Surface Mine Soil and Native Tallgrass Prairie Soil on Growth of Prairie Grasses.

Abstract

Soil compaction and competition from non-native forages preclude

reestablishment of native deciduous forest communities on reclaimed calcareous

surface mined land on the Allegheny Plateau. Establishing tallgrass prairie

communities may increase structural and functional qualities of these lands.

Arbuscular mycorrhizal fungi (AMF) are critical for establishment and survival of

warm-season prairie grasses, especially when soil phosphorus (P) is low. It is unclear

if AMF associated with the mine soil vegetation would be effective in supporting

warm-season prairie grasses. Studies were conducted to determine if resident AMF are

as affective as native tallgrass prairie AMF for prairie grasses on reclaimed mine soil,

and to determine how the prairie grasses respond to AMF over a range of P levels.

Soil AMF inoculum was prepared from Claridon (CL) tallgrass prairie soil near

Marion, OH, and from a 30-year old reclaimed surface mine soil, now part of the

Wilds (WL), near Cumberland, OH. Growth of four prairie grasses colonized with CL

and WL AMF were compared with non-colonized controls, at three P levels, in a 16-

week glasshouse study. The WL AMF were as effective as CL AMF for all grasses

88

suggesting a lack of effective AMF would not likely be a barrier for prairie grass

establishment. Soil P did not reduce AMF colonization, but did reduce AMF efficacy

at higher levels. Big bluestem (Andropogon gerardii Vitman) and Indiangrass

(Sorghastrum nutans (L.) Nash) benefited most from AMF. Tall dropseed

(Sporobolus asper (Michx.) Kunth) and slender wheatgrass (Elymus trachycaulus

(Link) Gould ex Shinners) growth at low P was not different between AMF and non-

AMF plants, indicating a facultative association with AMF for each species. Growth

of slender wheatgrass without AMF exceeded growth with AMF at higher levels of P.

Keywords: Calcareous mine soil, land restoration, arbuscular mycorrhizal fungi,

prairie grass establishment.

Introduction

Surface coal mining negatively impacts landscapes by altering soil structure

and chemistry, and by disturbing or diminishing beneficial soil organisms such as

AMF. During mining, vegetation is removed so that topsoil can be pushed aside in

stockpiles until post-mining reclamation. The removal of topsoil and subsequent

stockpiling disrupts AMF symbiosis, reduces inoculum potential, and alters species

composition of the AMF community (Gould and Liberta, 1981; Waaland and Allen,

1987). Revegetation of mined land is a critical component of reclamation but a

reduced or altered AMF presence in the mine soil may inhibit establishment of desired

species.

89

The role of AMF is critical in restoring reclaimed mine soils (Lambert and

Cole, 1980; Waaland and Allen, 1987; Allen, 1989; Hetrick et al., 1994). Arbuscular

mycorrhizal fungi are symbiotic organisms that colonize root systems of most land

plants to gain organic carbon in return for minerals, primarily phosphorus, absorbed

and transferred to the plant by the fungi (Gerdemann, 1968; Rhodes and Gerdemann,

1975; Barrow et al., 1977; Smith and Smith, 1977; Cavagnaro, et al., 2005). This

symbiotic relationship is critical to the survival of many plant species, especially when

soil phosphorus is low (Brejda et al., 1993). In addition, AMF may affect plant

community composition and successional trajectories by differentially benefiting some

plants over others (Janos, 1980; Allen and Allen, 1984; Crowell and Boerner, 1988;

Allen and Allen, 1992; Gange et al., 1990; Gange et al., 1993; Hartnett et al., 1994;

Koske and Gemma, 1997; Gillespie and Allen, 2006).

While AMF symbiosis is common and occurs in nearly every terrestrial

environment (Smith and Smith, 1997), differences in the effectiveness of AMF occurs

over the landscape and with different management histories (Scullion et al., 1998;

Gillespie and Allen, 2006). Strains of AMF from infertile soils are shown to be more

effective at phosphorus transfer to plants than AMF from fertile soil (Henkel et al.,

1989). Greater effectivity has been found in AMF from zinc contaminated soil as well

other stressful habitats (Shetty et al., 1995; Thorne et al., 1998). These studies suggest

that in harsh, low-nutrient habitats, there is selection for superior AMF strains.

Furthermore, a certain degree of host-plant-specificity occurs between AMF and host

plants (Zhu et al., 2000; Ronsheim and Anderson, 2001; Bevor, 2002; Sanders, 2003).

90

This suggests that AMF specific to undisturbed native communities might be best

adapted and more effective to associated plants than introduced or non-native AMF.

Conversely, plants introduced into new environments might benefit less from the

indigenous AMF than would plants long established in those environments.

In southeast Ohio, surface coal mining has converted nearly 80,000 hectares of

native deciduous forest land to non-native forage grassland (Kaster and Vimmerstedt,

1996). Since 1972, reclamation laws have required the overburden to be contoured to

approximate the original landscape form, and the stockpiled topsoil to be spread over

the newly constructed landscape. However, revegetation need not necessarily include

native species if preapproved plans state otherwise (SMCRA 1977). In place of the

native forest species, reclaimed mined land has been seeded with non-native forage

species such as tall fescue (Festuca arundinaceae Schreb.), Kentucky bluegrass (Poa

pratensis L.), and bird‘s-foot trefoil (Lotus corniculatus L.), which establish easily and

tolerate soil compaction caused by reclamation procedures. These cool-season forages

have established a thick ground cover important for controlling erosion and have

likely maintained AMF across the landscape, but, they are competitive and have

inhibited development of the native vegetation and diversity.

Replacing the non-native forage complex with regionally native prairie species

is one alternative for increasing biodiversity as a first step in restoring ecosystem

function on reclaimed mine sites that are incapable of supporting native forest species.

Tallgrass prairies are native to parts of Ohio and may represent a diverse set of species

that could enhance the functional quality of the mined land (Transeau, 1935; Sala et

91

al., 1988; Klips, 2004). However, it is unclear if AMF associated with the cool-season

forage species currently growing on the reclaimed mine land would be effective in

supporting the tallgrass prairie vegetation. Warm-season tallgrass species are more

dependent on AMF than cool-season grasses (Loree and Williams, 1987; Hetrick et

al., 1991; Brejda et al., 1993; Noyd et al, 1995) and problems with host specificity or

effectivity could delay or limit their establishment (Cavender and Knee, 2006).

This research compares the infective and effective potential of AMF collected

from a central Ohio tallgrass prairie remnant with AMF from reclaimed mine soil on

growth of four native tallgrass prairie grasses. The grass species evaluated are big

bluestem (Andropogon gerardii Vitman), Indiangrass (Sorghastrum nutans (L.) Nash),

tall dropseed (Sporobolus asper (Michx.) Kunth), and slender wheatgrass (Elymus

trachycaulus (Link) Gould ex Shinners). Slender wheatgrass is a cool-season grass

while big bluestem, Indiangrass and tall dropseed are all warm-season grasses; all four

occur throughout the central grassland region of North America, including tallgrass

prairies (Hitchcock, 1971). The reclaimed mine soil in question has supported a non-

native forage complex of three species for 30 years. The tallgrass prairie remnant

contains 177 plant species including the signature tallgrass prairie grasses big

bluestem, Indiangrass, tall dropseed, little bluestem (Schizachyrium scoparium

(Michx.) Nash), and switchgrass (Panicum virgatum L.) (Klips, 2003; Klips, 2004).

The objectives of this research were 1) to determine if AMF associated with mine soil

vegetation are as affective as native tallgrass prairie AMF in supporting prairie grass

growth on reclaimed mine soil, and 2) to determine how these prairie grasses respond

92

to each AMF in relation to a range of soil phosphorus levels. The goal of these studies

was to identify growth responses of prairie grasses to AMF and phosphorus that would

aid in developing strategies to increase biodiversity and ecosystem function on

compacted reclaimed mine soil.

Methods

AMF sources and pot culture

Sources of AMF were the Claridon (CL) tallgrass prairie remnant near Marion,

Ohio, and the Wilds (WL), a reclaimed surface mined area near Cumberland, Ohio.

The CL site is a 2.2 ha linear remnant owned by the CSX Railroad and is overseen by

the Marion County Historical Society (Klips, 2003; Klips 2004). The WL site is

located on land that had been surface mined in the early 1980‘s, and was once part of

the Muskingum Mine, then owned and mined by Central Ohio Coal Company, but

was donated in 1986 to The International Center for the Preservation of Wild Animals,

Inc. (the Wilds). The area is part of the Allegheny Plateau of southeast Ohio, which


Appalachian Mountain Range.

Approximately 35 liters of soil were collected from each site during

September, 2005. Soil from the surface 20 cm was collected from 15-20 randomly

selected locations at each site using a metal spade shovel. At the CL location, samples

were collected along side of established prairie grasses, including big bluestem and

Indiangrass, to be sure that grass roots containing AMF would be included. At the

93

WL location, soil was collected from an area supporting non-native forage grasses and

legumes including Kentucky bluegrass, tall fescue, and birds-foot trefoil. These

species were found throughout the reclaimed mined area. Pot cultures of each AMF

source were prepared by mixing soil from each location 1:1 by volume with silica

sand in a portable cement mixer. The soil/sand mix was poured into 3.8-l plastic

nursery containers and sown with white clover (Trifolium repens L.). The containers

were placed on benches in a 20-27º C glasshouse with artificial lighting 12 hr day-1

.

The pot cultures were watered daily without fertilizer for 10 months. Inoculum was

prepared by chopping up soil and roots from each pot, discarding course roots and

tops, then mixing all soil and fine roots together for each AMF source.

Sterile soil to be used as growing medium was prepared by mixing topsoil

collected from the surface 20-cm at the Wilds with silica sand, 1:1 by volume, in a

portable cement mixer. The soil/sand mix was then steamed for 5 hr at 100º C. The

sterile soil was rested in plastic bins for 21 d at 20º C prior to use in the experiment.

Soil from each pot culture, the sterilized growing medium soil, and original WL

topsoil were analyzed by the Service Testing and Research Laboratory (STAR lab),

The Ohio State University/Ohio Agricultural Research and Development Center,

Wooster, OH (Table 14). Identification of AMF species was not attempted for this

study.

Experiment establishment and design

Experimental units consisted of individual grass seedlings growing in 660-cm3

pots (D40 Deepot®, Stuewe and Sons, Inc., Corvallis, OR) containing 500 cm3 sterile

94

growing medium soil plus one of four AMF inoculum treatments, and one of three

phosphorus (P) levels. Inoculum treatments included 100 cm3 of CL or WL pot-

culture soil, or 100 cm3 of sterilized CL (CLS) or sterilized WL (WLS) pot culture

soil. Sterilized inoculum soil was added to the non-AMF pots to control for possible

fertilizer effects from adding pot culture soil to the AMF pots. The sterilized soil was

prepared by autoclaving 8 l of each pot culture soil for 70 min at 130º C, and then

resting the soil in plastic bags at 40º C for 96 h.

Soil P levels of 5, 12, and 24 mg kg-1

were established by mixing 0.0, 0.1, and

0.3 g triple super phosphate (0-45-0) (Bonide Products Inc., Oriskany, NY) per pot,

respectively. Calculations were based on the recommendation that 10 mg kg-1

P is

required to increase available soil P 1 mg kg-1

(Dr. Donald Eckert, PhD, The Ohio

State University). Each pot was standardized for bacteria by adding 100 ml of sievate

corresponding to each particular AMF inoculum. The sievate for each inoculum was

prepared by mixing 1000 cm3 pot culture soil and 16 l water, allowing the slurry to

settle for a few seconds, and pouring the liquid and suspended matter through a 53-µm

sieve.

The experiment was set up as a randomized complete block with a factorial

arrangement of 4 levels of grass species, 4 levels of AMF source (CL, CLS, WL, and

WLS), and 3 levels of P (5, 12, and 24 mg kg-1

). Each treatment was replicated six

times. The four grass species (SPP) were ‗Bison‘ big bluestem, ‗Tomahawk‘

Indiangrass, ‗Revenue‘ slender wheatgrass, and tall dropseed. Big bluestem,

Indiangrass, and slender wheatgrass were purchased from Western Native Seeds,

95

Coaldale, CO USA, and tall dropseed was purchased from Oak Prairie Farm,

Pardeeville, WI USA. Seeds of each species were sown 10-20 per pot, and thinned to

leave a single seedling in each pot.

Pots were placed in trays and arranged so that live AMF treatments were

adjacent to non-AMF control pots to allow for paired-pot comparisons of AMF

sources. Trays were placed on a glasshouse bench in a randomized-block design, such

that each treatment was contained in each block to control for distance from the

cooling/heating source on one end and exhaust fan at the other. Artificial lighting was

set to maintain a minimum of 300 w m-2

16 h day-1

, and temperature ranged between

19-27º C.

Grass leaf and biomass measurements

At 4, 8, 12, and 16 weeks following germination, the numbers of living and

dead leaves were counted on each plant. To reduce confusion in successive censuses,

dead leaves were removed at each census and stored for later biomass measurement.

At the end of the 16-week experiment, plants were destructively harvested to assess

above- and belowground biomass. Culms and leaves were clipped at the soil surface

and put in paper bags along with dead leaves from earlier censuses. Roots were

washed to remove the soil and then bagged separately from shoots. Biomass samples

were dried at 55º C for a minimum of 96 h, and then weighed. Three small sub-

samples were cut fresh from each root system to assess AMF colonization. The root

sub-samples were approximately 10x25 mm each and cut from the top, middle, and

96

bottom third of the root length. Root sub-samples were stored in a 48% ethanol

solution until being processed for AMF evaluation.

AMF colonization assessment

Root samples were cleared and stained according to a modified Phillips and

Hayman (1970) procedure. During processing, root samples from each plant were

contained in 28x5-mm tissue processing cassettes (Canemco Inc., Quebec, Canada).

Roots were cleared in 10% KOH solution and autoclaved at 130º C for 10 min, and

then acidified in a 1% HCL solution for 20 min at room temperature. Roots were

stained in 0.05% Trypan blue staining solution containing 1:2:1 distilled water, lactic

acid, and glycerin, and autoclaved for 7 min at 130 º C. Following staining, roots

were rinsed in tap water and stored in plastic Petri dishes covered with a 1:1 solution

of distilled water and glycerin and kept in a 40º C cooler. Colonization was assessed

using a gridline method (Newman, 1966; Giovannetti and Mosse, 1980). Roots

bisecting gridlines scored on the bottom of a Petri dish and spaced 13 mm apart, were

designated either colonized or not colonized. For each root sample, the first 50 roots

observed were scored colonized if the root segment crossing a gridline contained

hyphae, arbuscules, or vesicles. Percent colonization was calculated by dividing the

number colonized by 50, then multiplying by 100. The root sample was then dried at

55º C and weighed, and the dry weight was added back to the total root biomass.

97


Data were analyzed using PROC GLM in SAS/STAT® software (SAS, 2007)

and significance was accepted at α=0.05. Main effects were SPP, AMF, and P. Post-

hoc comparisons were made using protected Fisher‘s LSD test and differences were

accepted only if the P-value calculated by PROC GLM was equal or less than 0.05

(Milliken and Johnson, 1984). Dependent variables were leaf number, shoot, root, and

total biomass, root-to-shoot ratio (RSR), difference between AMF and non-AMF for

shoot (SDIFF), root (RDIFF), and total biomass (TDIFF), and AMF root colonization

percent. Difference in biomass was calculated as a separate continuous random

variable for each paired-pot comparison (Zar, 1999). Benefit from AMF inoculation

was indicated by a positive outcome after subtracting the non-AMF value from the

AMF value. Analysis of colonization percent only included the inoculated treatments

in order to accurately the level of infectivity of each inoculum.


AMF colonization as affected by soil P, inoculum, and grass species

The reclaimed mine soil used in this experiment measured 12 mg kg-1

P, 3768

mg kg-1

calcium (Ca), and pH of 7.3 (Table 14). These values indicate a calcareous

soil with limited available P. Mixing the soil with silica sand reduced the available P

to 5 mg kg-1

creating a soil critically low in P for plant growth and a soil environment

conducive to test and compare the efficacy of AMF strains. In our study, we found

that neither AMF source nor P concentration had any effect on colonization percent

98

when averaged over all other factors (Table 15). Grasses inoculated with CL averaged

52.3% AMF colonization, whereas grasses with WL averaged 53.1%. This indicates

that both AMF cultures were equally accepted by the host grasses in this study.

Grasses grown at 5, 12, and 24 mg kg-1

P averaged 55.6, 53.7, and 48.9% AMF

colonization, respectively, but were not different (Table 15). A number of studies

have shown that AMF colonization is reduced by higher soil P (Mosse, 1973; Schubert

and Hayman, 1986; Sainz and Arines, 1988; Al-Karaki and Al-Omoush, 2002), but

that was not evident in this study. If plants are able to obtain P with their own root

systems, then the symbiosis would clearly be less beneficial. However, there is often

no clear relationship between colonization percent and P uptake or plant growth

response (Lioi and Giovannetti, 1987; Sanders and Fitter, 1992; Mohammad et al.,

1995), meaning that efficacy in not necessarily related to the magnitude of

colonization.

Colonization did differ by species as tall dropseed and slender wheatgrass had

the highest percentages with 69.9 and 54.7%, respectively (Table 15). Big bluestem

and Indiangrass had lowest colonization with 51.0 and 35.5%, respectively. It is

known that warm-season grasses tend to be more dependent on AMF, especially when

P is limited, than cool-season grasses. Cool-season grasses tend to have finer root

systems that are better suited to gaining P from the soil, while warm-season grasses

tend to have more coarse root systems (Hetrick et al., 1991), which is why the high

abundance of AMF colonization in slender wheatgrass roots was unexpected. Big

bluestem is known to be very dependent on AMF (Hetrick et al., 1986) and is a

99

dominant species in tallgrass prairies across North America. The mycorrhizal status of

tall dropseed has not been reported, but a related species, Sporobolus heterolepis was

found to be mycorrhizal (Dhillion, 1992).

AMF and soil P effect on plant growth

Overall, biomass production was greatest for slender wheatgrass, which

averaged 1.5 g shoot biomass, 1.3 g root biomass, and 2.8 g total biomass plant-1

(Table 16). Big bluestem root biomass averaged 1.5 g plant-1

and was not different

from slender wheatgrass; however, big bluestem averaged only 0.7 g shoot biomass

plant-1

. The difference in allocation of resources between these species was reflected

in the RSR as slender wheatgrass averaged 0.9 while big bluestem averaged 2.4 (Table

16). Indiangrass and tall dropseed each produced 1.8 g total biomass plant-1

, which

was less than either slender wheatgrass or big bluestem. However, the RSR differed

between Indiangrass and tall dropseed as Indiangrass averaged 1.8, and tall dropseed

averaged 0.7. This indicates that big bluestem and Indiangrass appear to direct more

resources, proportionately, to root growth during seedling establishment, compared

with slender wheatgrass and tall dropseed.

Response to soil P concentration was predictable as an increase in P resulted in

an increase in biomass production (Table 16). However, this occurred only for shoot

biomass, as biomass increased from 0.6 to 1.0 to 1.3 g plant-1

as P increased from 5 to

12 to 24 mg kg-1

, respectively. Root biomass increased from 0.7 to 1.4 g plant-1

at 5

and 12 mg kg-1

P, respectively, but did not increase at 24 mg kg-1

P. This resulted in a

100

decline in the RSR from 1.6 to 1.3 between 12 and 24 mg kg-1

P as the increased shoot

growth at the higher P was not matched by a corresponding increase in root growth.

This is likely a result of space limitation in the pots and not a lack of response to

increased P, as pots with the highest P level were densely packed with roots when

harvested. However, total biomass was greatest at 24 mg kg-1

P compared with the 5

and 12 mg kg-1

levels (Table 16).

AMF effectivity in paired-pot comparison

In comparing the effectiveness of the AMF cultures, a paired-pot arrangement

was used to examine the difference in biomass accumulation between colonized and

non-colonized plants. By subtracting the biomass of a non-AMF plant from an

adjacent AMF-plant for each component (shoot/root/total), new variables were created

that if positive, indicated AMF benefit, and if negative, indicated AMF detriment.

Findings in the GLM analysis indicated that P and SPP had the greatest influence on

all three difference variables (Table 17). The F-values associated with P were 38.9,

29.4, and 51.2 for shoot difference (SDIFF), root difference (RDIFF), and total

differences (TDIFF), respectively. For SPP, F-values were 15.5, 19.2, and 25.5 for the

same variables, respectively. Furthermore, the F-values were highly significant for P

and SPP, each having P-values <0.0001 for all variables. In contrast, the F-values for

AMF source were only 6.0 for SDIFF, 0.6 for RDIFF, and 0.2 for TDIFF.

The interaction between AMF and P was significant for RDIFF (P<0.0001)

and TDIFF (P<0.0001), but not for SDIFF (P=0.0643) (Table 17). For SDIFF, the

101

response to AMF to P was nearly identical with each AMF source (Table 18), which

would explain the lack of interaction. For SDIFF, the outcome was positive at 5 mg

kg-1

P, averaging 0.22 for CL and 0.24 for WL, meaning that the grasses benefitted

similarly from each culture. However, at 12 mg kg-1

P the outcome was negative for

both AMF sources averaging -0.38 for CL and -0.31 for WL, suggesting AMF was

detrimental. At 28 mg kg-1

P, the outcomes for SDIFF were also negative and

averaged -0.81 for CL and -0.34 for WL (Table 18). Furthermore, within each AMF

source, SDIFF decreased with each increase in P. In contrast, RDIFF outcomes were

similar between 5 and 12 mg kg-1

P with CL and WL averaging -0.06 and -0.34,

respectively (Table 18). The RDIFF was positive only at 5 mg kg-1

P with the WL

source, which averaged 0.30, but averaged -1.50 and -0.76 at the 12 and 24 mg kg-1

P,

respectively. Similarly, TDIFF was positive only at the 5 mg kg-1

P and averaged 0.15

for CL and 0.53 from WL.

The interaction between SPP and P was also significant for SDIFF, RDIFF,

and TDIFF. A breakdown of this interaction shows that AMF benefited only big

bluestem and Indiangrass at the lowest P level (Table 19). Big bluestem averaged

0.41, 1.01, and 1.42 for SDIFF, RDIFF, and TDIFF, respectively, at 5 mg kg-1

, but

each increase in P yielded negative values. At 24 mg kg-1

P, SDIFF, RDIFF, and

TDIFF averaged -0.92, -1.19, and -2.11, respectively. Indiangrass responded similarly

as SDIFF, RDIFF, and TDIFF were positive, 0.55, 0.89, and 1.44, respectively, at 5

mg kg-1

P, but were negative at the higher P levels. However, for Indiangrass, no

102

decrease in these variables occurred between 12 and 24 mg kg-1

P. Neither species

appeared to benefit from AMF at the higher P levels.

In contrast, no apparent benefit from AMF at any concentration of P could be

seen for tall dropseed or slender wheatgrass (Table 19). Tall dropseed had positive

SDIFF outcomes of 0.09 and 0.01 for 5 and 24 mg kg-1

P, but these were not different

from the -0.13 outcome recorded at 12 mg kg-1

P. Furthermore, differences were not

found between P concentrations for RDIFF or TDIFF, and all outcomes were negative.

Likewise, slender wheatgrass had negative values for SDIFF, RDIFF and TDIFF at all

P concentrations; however, SDIFF and TDIFF at 5 mg kg-1

P were less negative than

at 12 and 24 mg kg-1

P. This suggests that response of slender wheatgrass to AMF is

more negatively affected by high P concentrations than is tall dropseed.

The interaction between SPP and P was evident also in the number of leaves

produced during the 16-week experiment (Figure 4). Leaf production with 5 mg kg-1

P was greater for AMF big bluestem, Indiangrass, and tall dropseed at each census.

Indiangrass appeared to benefit more from AMF than either big bluestem or tall

dropseed as the difference in leaf count between colonized and non-colonized plants

was significantly greater for Indiangrass. In contrast, at 5 mg kg-1

P, slender

wheatgrass with AMF produced slightly more leaves only at 4 weeks.

At 12 and 24 mg kg-1

P, benefit from AMF was less evident (Figure 4). Big

bluestem with AMF had slightly greater leaf counts at the 4, 12, and 16-week censuses

with 12 mg kg-1

P, but only at 8 weeks with 24 mg kg-1

P. At 4 weeks with 12 mg kg-1

P, Indiangrass plants with AMF had slightly greater leaf counts than non-AMF plants.

103

Leaf counts for tall dropseed with AMF at the higher P levels were not different from

non-AMF plants at any census. In contrast slender wheatgrass with AMF produced

fewer leaves than non-AMF plants at all four censuses at the 12 and 24 mg kg-1 P

levels. Furthermore, more leaves were produced by non-AMF slender wheatgrass

with 24 mg kg-1

P than for any other grass or treatment level, indicating superior

growth response to added soil P.

Conclusions

Results of this research indicate that AMF associated with reclaimed mine soil

are not likely a barrier for establishing tallgrass prairie species on the reclaimed mine

soil. Colonization levels are similar between the two AMF inoculums, and plant

growth response to each AMF is similar at all three P levels. This would also suggest

that host specificity is not an issue even though the mine soil AMF have been

associated with non-native forage species for 30 years. It appears that poor soil

conditions of the mine soil, i.e. compacted calcareous soil with low available

phosphorus, may have selected for AMF equally effective in supplying P as AMF

from undisturbed tallgrass prairie.

Big bluestem and Indiangrass responded to AMF and P as expected according

to previous research (Hetrick et al., 1986). Both species benefited from AMF when

soil P was low, and showed less benefit as P increased. Both of these two grasses

allocated more resources to roots than aboveground tissue, which is important for

access to nutrients and water during periods of stress. Harris (1967) determined that

104

competitive success of non-native downy brome (Bromus tectorum L.) was due to its

ability to establish a deep root system during autumn and winter when the native

bluebunch wheatgrass (Pseudoroegneria spicata (Pursh) A. Löve) was dormant.

During spring, a downy brome infestation depleted soil moisture before the

wheatgrass was able to complete its reproductive cycle. The dominance of big

bluestem and Indiangrass in tallgrass prairies is likely due to their ability to establish

deep root systems over time, as well as their association with AMF when soil P is

limited.

Tall dropseed and slender wheatgrass both appear facultative in their response

to AMF. Tall dropseed is a warm-season prairie grass, but seems to respond to AMF

and P similarly to facultative cool-season grasses. Greater production of aboveground

biomass than root biomass, and low dependence on AMF, would suggest that tall

dropseed can quickly establish following disturbance in habitats where P may be

limiting. Slender wheatgrass forms association with AMF but is clearly not dependent

on AMF and is able to access P when soil levels are low, and can be very productive

when soil P is higher. Tall dropseed and slender wheatgrass both appear to establish

well when P is low, with or without AMF and would be useful in early establishment

of a tallgrass prairie community on reclaimed mine soil. Big bluestem and Indiangrass

are more dependent on AMF and benefited from the mine soil AMF in this study.

Mine soil AMF. Establishing native prairie species on reclaimed compacted mine soil

would not likely be limited by the effectivity of the mine soil AMF.

105

Table 14. Soil properties of reclaimed mine and AMF inoculum soil used to

compare growth of prairie grasses with different concentrations of phosphorus

(P) and different sources of arbuscular mycorrhizal fungi (AMF).

Mine

2

topsoil

(0-20 cm)

Sterile mine

soil/sand

mix (1:1)

AMF3

Soil Parameter1 CL WL

pH 7.3 7.3 7.7 7.9

P (mg kg-1

) 12 5 < 1 7

K (mg kg-1

) 161 80 77 41

Ca (mg kg-1

) 3768 1722 1345 1262

Mg (mg kg-1

) 321 198 235 198

1 Soil analyzed by STAR lab, Wooster, OH. P analyzed with Bray P1 method; K,

Ca, and Mg analyzed with ammonium acetate extract method.

2 Soil collected from reclaimed surface mine land near Cumberland, OH currently

part of the Wilds (The International Center for the Preservation of Wild

Animals, Inc.).

3 CL collected from Claridon tallgrass prairie remnant near Marion, OH; WL

collected from the Wilds reclaimed mine soil supporting non-native forage

grasses. AMF inoculum soil prepared as pot-cultures containing a 1:1 mix of

soil and silica sand.

106

Table 15. Percent colonization by arbuscular mycorrhizal fungi (AMF) as

affected by AMF sources, soil phosphorus concentration (P), and grass

species (SPP) in a 16-week glasshouse experiment.

Parameter Colonization1

AMF (%)

CL 52.3 a

WL 53.1 a

P-value (0.7917)

P (mg kg-1

)

5 55.6 a

12 53.7 a

24 48.9 a

P-value (0.2271)

SPP

Big bluestem 51.0 bc

Indiangrass 35.5 c

Tall dropseed 69.9 a

Slender wheatgrass 54.7 ab

P-value (0.0226)

1 Colonization percents reflect only AMF-inoculated plants. Non-inoculated

plants had 0% AMF colonization.

107

Table 16. Biomass production as affected by grass species (SPP) and three levels

of soil phosphorus (P) in a 16-week glasshouse experiment.

Dependent variables1

Main effects Shoot Root Total RSR2

SPP --------------- (g dry weight) -------------- --- (g/g) ---

Big bluestem 0.7 c 1.5 a 2.2 b 2.4 a

Indiangrass 0.7 c 1.1 b 1.8 c 1.8 b

Tall dropseed 1.1 b 0.7 c 1.8 c 0.7 c

Slender wheatgrass 1.5 a 1.3 ab 2.8 a 0.9 c

P-value <0.0001 <0.0001 <0.0001 <0.0001

P

5 mg kg-1

0.6 c 0.7 b 1.4 c 1.5 ab

12 mg kg-1

1.0 b 1.4 a 2.4 b 1.6 a

24 mg kg-1

1.3 a 1.4 a 2.7 a 1.3 b

<0.0001 <0.0001 <0.0001 0.0231

1 P-values represent the probability of differences within each dependent variable

for each main-effect. Differences within each variable are determined using

protected Fisher‘s LSD (α=0.05), and are designated by lower-case letters.

2 Root to shoot ratio (RSR) calculated by dividing root weight by shoot weight.

108

Table 17. Analysis of variance table (PROC GLM) for the full model testing the

difference in biomass production for grass species (SPP) colonized with

arbuscular mycorrhizal fungi (AMF) and non-AMF-inoculated plants. Dependent

variables shoot difference (SDIFF), root difference (RDIFF), and total difference

(TDIFF) were produced by subtracting biomass of non-AMF plants from AMF

plants in a paired-pot glasshouse experiment examining the effects of AMF

source and P on growth of prairie SPP grown in sterilized mine soil.

SDIFF RDIFF TDIFF

Model DF F value P > F F value P > F F value P > F

REP1 5 0.4 0.8753 2.2 0.0601 1.2 0.3225

SPP 3 15.5 <.0001 19.2 <.0001 25.5 <.0001

AMF 1 6.0 0.0162 0.6 0.4294 0.2 0.6244

P 2 38.9 <.0001 29.4 <.0001 51.2 <.0001

SPP *AMF 3 0.7 0.5354 2.8 0.0461 2.8 0.0428

SPP*P 6 3.9 0.0013 3.3 0.0053 5.2 <.0001

AMF*P 2 2.8 0.0629 15.6 <.0001 13.1 <.0001

SPP*AMF*P 6 0.5 0.8391 0.7 0.6449 0.9 0.5276

TOTAL 141

1 REP refers to replicate.

109

Table 18. Interaction of arbuscular mycorrhizal fungi (AMF) from the Claridon

tallgrass prairie remnant (CL) and the Wilds reclaimed calcareous mine soil

(WL) in Ohio and soil phosphorus concentration (P). Values represent the

difference between AMF-colonized and non-AMF-colonized grasses1 in a 16-

week glasshouse experiment measuring shoot (SDIFF), root (RDIFF), and total

biomass difference (TDIFF).

AMF P SDIFF RDIFF TDIFF

(mg kg-1

) ------------------ (g difference) -------------------

CL 5 0.22 a -0.06 a 0.15 a

12 -0.38 b -0.34 a -0.73 b

24 -0.81 c -1.23 b -2.03 c

P-value (<0.0001) (0.0002) (<0.0001)

WL 5 0.24 a 0.30 a 0.53 a

12 -0.31 b -1.50 c -1.81 b

24 -0.34 c -0.76 b -1.10 b

P-value (0.0013) (<0.0001) (<0.0001)

1 Grass species are big bluestem, Indiangrass, tall dropseed, and slender wheatgrass.

110

Table 19. Interaction of grass species (SPP) and soil phosphorous concentration (P)

on the difference in biomass between grasses colonized with arbuscular

mycorrhizal fungi (AMF) and non-AMF colonized grasses in a 16-week

glasshouse experiment measuring shoot (SDIFF), root (RDIFF), and total

biomass difference (TDIFF).

Grass species P SDIFF RDIFF TDIFF

(mg kg-1

) ----------------- (g difference) -------------------

Big bluestem 5 0.41 a 1.01 a 1.42 a

12 -0.27 b -0.34 b -0.60 b

24 -0.92 c -1.19 c -2.11 c

P-value (<0.0001) (<0.0001) (<0.0001)

Indiangrass 5 0.55 a 0.89 a 1.44 a

12 -0.16 b -0.64 b -0.80 b

24 -0.35 b -0.49 b -0.83 b

P-value (<0.0001) (0.0003) (<0.0001)

Tall dropseed 5 0.09 a -0.38 a -0.29 a

12 -0.13 a -0.94 a -1.07 a

24 0.01 a -0-79 a -0.79 a

P-value (0.4259) (0.1029) (0.0936)

Slender wheatgrass 5 -0.11 a -0.84 a -0.95 a

12 -0.82 b -1.79 a -2.60 b

24 -0.98 b -1.47 a -2.45 b

P-value (<0.0001) (0.1927) (0.0116)

111

Ind

iang

rass

(lea

ves

pla

nt-1

)

0

10

20

30

40

Tal

l d

rop

seed

(lea

ves

pla

nt-1

)

0

10

20

30

40

(weeks)

5 mg kg-1

P

0 4 8 12 16

Sle

nd

er w

hea

tgra

ss

(lea

ves

pla

nt-1

)

0

20

40

60

80

(weeks)

12 mg kg-1

P

0 4 8 12 16

(weeks)

24 mg kg-1

P

0 4 8 12 16

Big

blu

este

m

(lea

ves

pla

nt-1

)

0

10

20

30

40

**

* *

**ns

* *

ns*

ns ns

***

***

******

*

ns

nsns

ns

ns

nsns

ns*

**

*ns

nsns

ns

ns

nsns

*ns

ns

ns

*

**

*

*

**

**

**

**

Figure 4. Affect of arbuscular mycorrhizal fungi (AMF) on number of leaves

produced by prairie grasses at three concentrations of soil phosphorus (P) level during

a 16-week glasshouse experiment. Solid lines and circles (●—●) represent AMF

colonized; dotted lines and open circles (○····○) represent non-AMF colonized plants.

P-values comparing AMF and non-AMF leaf counts are signified as follows:

= 0.05<P<0.001, = 0.001<P<0.0001, = P<0.0001, and ns = P>0.05.

112

Chapter 6: Effect of soil compaction and arbuscular-mycorrhizal fungi on tall fescue,

big bluestem and eastern gamagrass seedling growth.

Abstract

Soil compaction on reclaimed surface-mined lands in Appalachia is a barrier to

restoration of deciduous forests native to the region. Non-native tall fescue (Festuca

arundinaceae Schreb.) was planted because it establishes quickly and tolerates soil

compaction. Establishing robust native grasses such as eastern gamagrass (Tripsacum

dactyloides L.) and big bluestem (Andropogon gerardii Vitman) could help ameliorate

the compaction by rooting deep into the profile, while increasing biodiversity. These

warm-season grasses are native to the tallgrass prairie regions of North America and

are major components of prairie communities. Warm-season grasses tend to have

coarse roots and are more dependent on arbuscular-mycorrhizal fungi (AMF) than

cool-season grasses. The symbiosis with AMF increases survival in soils with low

available phosphorus. A10-week glasshouse study was conducted to examine the

interaction between soil compaction and AMF source (reclaimed mine soil, native

tallgrass prairie soil, or none) on seedling growth of tall fescue, big bluestem, and

eastern gamagrass. Growth of tall fescue with AMF was not different from that with

no AMF. Big bluestem and eastern gamagrass were highly dependent on AMF, but

differences were not found between mine soil (WL) and prairie (CL) AMF.

113

Colonization was greatest for eastern gamagrass and exceeded 70% with either WL or

CL. Big bluestem and tall fescue colonization was 1.75 and 3 times greater with WL

AMF than CL AMF, respectively. Tall fescue produced 3 times greater shoot and root

biomass than eastern gamagrass and 6 times greater than big bluestem when colonized

with AMF. The highest compaction level reduced tall fescue root biomass by 50%

and shoot biomass by 40%. Big bluestem root biomass was reduced 60% and shoot

biomass 50% by the highest compaction. Eastern gamagrass shoot and root biomass

was not affected by increasing compaction. Eastern gamagrass is slower to establish

than tall fescue but tolerates compaction, which would be beneficial in compacted

mine soil.

Keywords: Soil compaction, arbuscular-mycorrhizal fungi, reclaimed mine soil,

prairie grasses.

Introduction

Soil compaction is a major limiting factor in restoring native vegetation on

reclaimed surface mined land in the Appalachian coal mining region of the eastern

U.S. (Larson and Vimmerstedt, 1983; Kost et al, 1994; Zeleznik and Skousen, 1996;

Haering et al., 2004; Casselman et al., 2006). Bulk densities of up to 1.8 g cm-3

to

depths of at least 60 cm have been recorded following use of heavy equipment to

redistribute and contour overburden and stockpiled topsoil during reclamation

(Shrestha et al., 2009). Soil compaction limits available water and nutrients, reduces

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pore volume, restricts root elongation and development, reduces shoot biomass and

growth, and leads to anaerobic conditions that many plants are unable to tolerate

(Unger and Kaspar, 1994; Whalley et al., 1995; Bengough et al., 1997; Nadian et al.,

1997; Kozlowski, 1999).

Ameliorating soil compaction on reclaimed mine land is possible with deep

tillage (Sinnett et al., 2006; Croton and Ainsworth, 2007; Shrestha et al., 2009).

Tillage can reduce bulk density and improve water infiltration and has been used

extensively in agriculture; however, deep tillage requires heavy equipment and high

levels of energy input (Sinnett et al., 2006; Croton and Ainsworth, 2007) and disrupts

developing soil structure and beneficial micro-organism such as arbuscular-

mycorrhizal fungi (AMF) (Jasper et al., 1989; Lal, 1993). An alternative to tillage is

planting deep-rooting plant species that can penetrate and increase organic matter and

biological activity in compacted regions of the profile. Roots that grow into

compacted soil create channels that increase water flow and are used by the roots of

other species to access soil otherwise not available because of the compaction (Meek

et al, 1990; Cresswell and Kirkegaard, 1995; Rasse and Smucker, 1998; Unger and

Kaspar, 1994; Williams and Weil, 2004). Tap rooted annual plants are often used in

agricultural systems; however, perennial species such as alfalfa may be more effective

because of long-term growth and deeper rooting characteristics. Using perennial

species would also eliminate the need for annual tillage, fertilization, and planting

associated with annual cropping.

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Tall fescue, a perennial non-native forage grass, has been planted extensively

on reclaimed surface-mined land in Appalachia (Casselman et al, 2006). Tall fescue is

very productive and tolerates mine soil compaction (Crews, 1984; Sulc et al., 2006)

and establishes within the five-year window mining companies have to establish a

vegetation cover on reclaimed surface-mined land. However, tall fescue productivity

declines over time on mine soil without organic amendments supplying nitrogen and

phosphorus, or legumes fixing nitrogen, (Roberts et al., 1988a). Furthermore, only

planting a few non-native forages on mine soil results in a low diversity landscape.

Establishing large-statured, robust prairie species as an alternative to low-

diversity forages, might improve soil conditions on reclaimed mine land over time.

Big bluestem (Andropogon gerardii Vitman) and eastern gamagrass (Tripsacum

dactyloides L.) are warm-season C4 grasses native to prairies in the Midwest and

southeastern U.S. (Hitchcock, 1971). Big bluestem is dominant in tallgrass prairies

and historically comprised as much as 90% canopy cover within its range (Weaver,

1931). Eastern gamagrass is a robust perennial relative of corn (Zea mays L.) and is

prevalent in the southern tallgrass prairies regions; however has declined over time

with overgrazing and tillage-based agriculture (Gilker et al., 2002). Furthermore,

eastern gamagrass is productive on compacted mine soil due to its ability to root

through compacted soil layers and tolerance to flooding (Clark et al., 1998: Gilker et

al., 2002; Krizek et al., 2003).

Warm-season prairie grasses tend to have coarser root systems than cool-

season grasses but thrive in soils with low available nutrients, especially phosphorus,

116

because of the symbiotic association with AMF (Daniels et al., 1988; Hetrick et al.,

1991; Brejda et al., 1993). The importance of AMF to plant survival is well

documented (Mosse, 1973; Gerdemann, 1968), but AMF also benefit plants growing

in compacted soil (Nadian et al., 1997; Miransari et al., 2007). Establishment of

warm-season prairie grasses on reclaimed mine soil may help ameliorate mine soil

compaction and increase community diversity, but only if the AMF associated with

the mine soil is effective and will benefit the prairie grasses. The objective of this

research is to compare the early growth of tall fescue with big bluestem and eastern

gamagrass in compacted soil in relation to AMF from reclaimed mine soil, native

tallgrass prairie soil, and soil with no AMF. The goal was to determine the

effectiveness of mine soil AMF relative to tallgrass prairie AMF, and to compare the

growth response of each grass species under varying levels of soil compaction, in a

glasshouse study.

Methods

AMF sources and pot culture

Sources of AMF were the Claridon (CL) tallgrass prairie remnant near Marion,

Ohio, and the Wilds (WL), a reclaimed surface mined area near Cumberland, Ohio.

The CL site is a 2.2 ha linear remnant owned by the CSX Railroad and is overseen by

the Marion County Historical Society (Klips, 2003; Klips 2004). The WL site is

located on land that had been surface mined in the early 1980‘s, and was once part of

the Muskingum Mine, then owned and mined by Central Ohio Coal Company, but

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was donated in 1986 to The International Center for the Preservation of Wild Animals,

Inc. (the Wilds). The area is part of the Allegheny Plateau of southeast Ohio, which


Appalachian Mountain Range.

Approximately 35 liters of surface topsoil were collected at each site during

September, 2005 from 15-20 randomly selected locations at each site using a metal

spade shovel to a depth of 20 cm. At the CL location, samples were collected along

side of established prairie grasses, including big bluestem and Indiangrass, so that

grass roots containing AMF would be included. At the WL location, soil was

collected from an area supporting non-native forage grasses and legumes including

Kentucky bluegrass (Poa pratensis L.), tall fescue, and birds-foot trefoil (Lotus

corniculatus L.). These species were found throughout the reclaimed mined area.

Pot cultures of each AMF source were prepared by mixing soil from each

location 1:1 by volume with silica sand in a portable cement mixer. The soil/sand mix

was poured into 3.8-l plastic nursery containers and sown with white clover (Trifolium

repens L.) as a host plant for the AMF (Liu and Wang, 2003). The clover was

inoculated with rhizobia to insure nitrogen fixation. The containers were placed on

benches in a 20-27º C glasshouse with artificial lighting 12 hr day-1

. The pot cultures

were watered daily without fertilizer for 10 months. Soil and fine roots were collected

from each pot, mixed 1:1 with sterile soil/sand mix, and repotted and seeded with

white clover and sideoats grama (Bouteloua curtipendula (Michx.) Torr.) for a second

pot-culture cycle. After 4 months, watering was ceased and the pots moved to an

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artificially-lighted 16-18º C room until the white clover and sideoats grama had wilted

and dried. Fine roots and soil were collected from each pot and then mixed together

for each AMF source and stored in plastic bags at 5º C for several days before being

used in the experiment.

Sterile growing medium was prepared by sieving topsoil collected from the

surface 20-cm at the Wilds through a 6-mm sieve. The sieved soil was then steamed

for 5 hr at 100º C and rested in plastic bins at 20º C prior to use in the experiment. The

sterilized mine soil and the WL and CL pot-culture soils were analyzed by the Service

Testing and Research Laboratory (STAR lab), The Ohio State University/Ohio

Agricultural Research and Development Center, Wooster, OH (Table 20).

Identification of AMF to species was not attempted for this study.

Experiment design and establishment

Experimental design was a randomized complete block with a factorial

arrangement of three grass species, three soil compaction levels, and three AMF

treatments; each complete block was replicated six times. Experimental units

consisted of individual plants growing in polyvinyl chloride (PVC) tubes measuring 8-

cm inside diameter by 30-cm deep. The inside of each tube was coated with copper

hydroxide (SpinOut® root growth regulator, American Hydrotech, Inc., Chicago, IL.)

to prevent roots from avoiding the compacted soil medium by growing down the side

of the tubes. Soil was mixed individually for each tube by measuring an appropriate

amount of sterile soil, on a dry weight basis, plus 120 cm3 AMF-inoculum soil, to

119

yield bulk densities (BD) of 1.0, 1.25 and 1.5 g cm-3

. The AMF control tubes were

filled only with sterile soil. Compaction levels were established by filling tubes in 5-

cm increments with a measured amount of soil to give the desired bulk density. A

solid-wood plunger with 5-cm reference lines, slightly smaller in diameter than the

PVC tubes, was pressed down on the soil while the bottom of tube was simultaneously

tapped on a concrete floor. Tapping continued until the appropriate reference line for

each depth lined up with the top of the tube.

Seeds of ‗Jesup MaxQ‘ tall fescue, ‗Bison‘ big bluestem and ‗Pete‘ eastern

gamagrass were germinated in Petri dishes and transplanted into each tube. After

transplanting, the soil surface was covered with a layer of vermiculite to prevent

drying and cracking. Each tube was then standardized for bacteria by adding 100 ml

of sievate corresponding to each particular AMF inoculum. The sievate for each

inoculum was prepared by mixing 1000 cm3 pot culture soil and 16 l water, allowing

the slurry to settle for a few seconds, and pouring the liquid and suspended matter

through a 53-µm sieve. Sievate from the WL inoculum was applied to the non-AMF

tubes. An endophyte-free tall fescue cultivar was used in this experiment to avoid

inhibition of AMF (Chu-Chou et al., 1992; Guo et al., 1992)

The 27 tubes in each block were randomly arranged in three rows of nine tubes

in a 43x122-cm wooden rack fitted into a 10x60x122-cm plastic tub. Each tube was

placed on a 10x8x10-cm block of floral foam (Aquafoam®, Syndicate Sales, Inc.,

Kokomo, IN). Holes were cut in the sides of the tubs 1 cm from the bottom for

drainage and the tubes were watered daily as needed without fertilizer. The tubs were

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placed on benches in a glasshouse under artificial lighting set to maintain a minimum

of 300 w m-2

16 h day-1

, and temperature set to range between 19-27º C. Block

establishment was staggered so that harvest of the plants would not occur at the same

time, and yet maintain an overall 10-week growing period. The experiment was

conducted from April through July 2008, to take advantage of increasing natural day

length, and moderate outside temperatures.

Plant growth and AMF measurements

At the end of the 10-week growth period, plant height (highest culm), number

of leaves and tillers, aboveground (shoot) biomass, root biomass, and AMF

colonization were measured. Shoots were clipped at the soil surface and soil was

washed from the roots. Shoots and roots were placed in separate paper bags and dried

at 55º C for a minimum of 96 hr, then weighed. Root to shoot ration (RSR) was

calculated by dividing root dry weight by shoot dry weight. Three small root sub-

samples 10x25 mm were cut from each root length to assess AMF colonization.

AMF colonization assessment

Root samples were cleared and stained according to a modified Phillips and

Hayman (1970) procedure. During processing, root samples from each plant were

contained in 5x28 mm tissue processing cassettes (Canemco Inc., Quebec, Canada).

Roots were cleared in 10% KOH solution and autoclaved at 130º C for 10 min, and

then acidified in a 1% HCL solution for 20 min at room temperature to improve

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staining. Roots were stained in 0.05% Trypan blue staining solution containing 1:2:1

distilled water, lactic acid, and glycerin, and autoclaved for 7 min at 130 º C.

Following staining, roots were rinsed in tap water and stored in plastic Petri dishes

covered with a 1:1 solution of distilled water and glycerin and kept in a 40º C cooler.

Colonization was assessed using a gridline method (Newman, 1966; Giovannetti and

Mosse, 1980). Roots were scored either colonized or not colonized when they

bisected gridlines on the bottom of a Petri dish spaced 13 mm apart. For each root

sample, only the first 50 roots were scored. Roots were scored colonized if the root

segment crossing a gridline contained hyphae, arbuscules, or vesicles. Percent

colonization was calculated by dividing the number colonized by 50, then multiplying

by 100.


Data were analyzed using PROC GLM in SAS/STAT® software (SAS, 2007)

and significance was accepted at α=0.05. Independent variables were grass species,

AMF inoculum, compaction level, and block. Dependent variables were leaf and tiller

count, shoot and root biomass, RSR, and percent AMF root colonization. All data

except AMF colonization were rank transformed (Iman et al., 1984) to address

normality and equal variance issues; AMF percent colonization data were arcsine

transformed. Post-hoc comparisons were made on transformed data using protected

Fisher‘s LSD test and differences were accepted only if the P-value calculated by

PROC GLM was equal or less than 0.05 (Milliken and Johnson, 1984). For

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presentation, actual means were used in place of ranks, and the sine of the arcsine

means were used to back-transform the colonization data.


Soil analysis

The calcareous mine soil used in this study had pH of 7.3, calcium (Ca)

content of 3768 mg kg-1

and available phosphorus (P) content of 12 mg kg-1

(Table

20). The basic pH and high Ca content is due to CaCO3 from limestone layers

unearthed during the mining process (Brant, 1964; Soil Survey Staff, 1996). In

calcareous soil, P becomes unavailable for plant uptake as it binds with Ca to form

calcium phosphate compounds (Brady, 1984; McBride, 1994). Availability of soil P

depends on factors including parent material, soil pH, temperature, total soil P, and

associated plant species (Pearson et al., 1940; Barrow et al., 1977; Wuenscher and

Gerloff, 1977; Roberts et al., 1988b; Jayachandran et al., 1989; Richardson et al.,

2009). Schubert and Hayman (1986) found that AMF was not a benefit to plant

growth when available soil P was greater than 50 mg kg-1

, suggesting that plant

growth may be limited without AMF in the 12 mg kg-1

P mine soil used in this

experiment.

AMF colonization

Percent AMF colonization was greatest for eastern gamagrass, but infectivity

was similar between the WL or CL inoculums (Table 21). Eastern gamagrass root

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colonization with WL averaged 76.2% and 73.3% with CL. The non-inoculated

eastern gamagrass roots were slightly colonized at 0.3%. In contrast, big bluestem had

55.9% root colonization with WL, which was greater than with CL at 31.7%, or 0%

for the non-inoculated plants. Tall fescue was 29.8% colonized with WL, but only

9.8% and 0% with CL and no AMF, respectively. Difference in colonization between

CL- and non-inoculated tall fescue plants was not found. Tall fescue was apparently a

better host for WL AMF than CL AMF. This is likely due to WL AMF being

associated with tall fescue and other cool-season grass on the reclaimed mine soil;

whereas, CL AMF from the tallgrass prairie remnant were primarily associated with

warm-season grasses including big bluestem. Host specificity has been shown for

other AMF cultures (Zhu et al., 2000; Ronsheim and Anderson, 2001; Bevor, 2002;

Sanders, 2003). However, big bluestem also had greater colonization by WL than CL

AMF, which would suggest that WL AMF is more infective than CL. Eastern

gamagrass showed no preference between the two AMF sources, but was clearly more

receptive to AMF than the other two grasses, especially tall fescue. Colonization

levels are not always correlated with efficacy; however, higher colonization may

reflect greater dependence, especially in coarse-rooted species such as eastern

gamagrass (Daniels et al., 1988; Hetrick et al., 1991). Compaction level had no

apparent effect on colonization percent in this study as none of the GLM model

components with compaction were significant at α=0.05 (data not shown).

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Effectivity of AMF on grass growth

Effectiveness of the AMF inoculums were evaluated, in part, based on grass

growth measured at the completion of the 10-week study. The three-way interaction

between grass species, AMF inoculum, and compaction level was not significant at

α=0.05 for any of the growth variables (data not shown); however, the interaction

between grass species and AMF was significant for all variables. When averaged over

compaction levels, plant growth was found to be similar for two AMF sources, but

differed between grass species (Table 22). For tall fescue, there were no differences in

growth between WL, CL, or no AMF inoculum treatments. Tall fescue averaged 77.4,

66.0, and 59.8 leaves plant-1

with WL, CL, and no AMF, respectively, while shoot

biomass averaged 6 g plant-1

for all three AMF inoculums, and root biomass averaged

3.8 for WL and 3.6 for both the CL and non-inoculated plants. Furthermore, the

similarity in shoot and root growth with each inoculum resulted in similar RSR values

as WL-inoculated tall fescue averaged 0.7 and CL and non-inoculated plants averaged

0.6. The similarity between AMF and non-AMF measurements supports other

findings that tall fescue‘s association with AMF is highly facultative, and in the

absence of AMF, can perform as well as AMF-colonized plants when soil is low in

available P (Lambert and Cole, 1980).

Growth measurements for big bluestem were similar between the WL and CL

inoculated plants for all variables, but, differences existed between AMF-inoculated

and non-inoculated plants (Table 22). Big bluestem produced 16.9 and 13.5 leaves

plant-1

, and 1.9 and 1.4 tillers plant-1

with WL and CL, respectively, but only averaged

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5.5 leaves and 1.0 tiller plant-1

with no AMF. Shoot biomass averaged 1.1 g plant-1

with either AMF, but only 0.01 g plant-1

with no AMF. Root biomass averaged 0.6

and 0.5 g plant-1

with WL and CL, respectively, and 0.1 g plant-1

with no AMF.

Similar results were found with eastern gamagrass as AMF-inoculated plants had

greater growth than non-inoculated plants (Table 22). Plants with AMF had 3 times as

many leaves and 4 times as many tillers as non-inoculated plants. Furthermore, shoot

biomass was 14 times greater and root growth 13 times greater for AMF-inoculated

plants. The one exception to this trend was in RSR as no difference was found

between AMF-inoculated (RSR=0.59) and non-inoculated eastern gamagrass

(RSR=0.67). In contrast, non-inoculated big bluestem had a RSR of 1.2, which was

greater than all others, and the only RSR greater than 1.0. Without AMF, big

bluestem was the only species to allocate more resources to root growth, likely to gain

access to P; however, overall biomass production with no AMF was nearly non-

existent.

The difference between AMF and non-AMF big bluestem and eastern

gamagrass was also evident in culm height (Table 22). Big bluestem culm height

averaged 49 cm with either WL or CL AMF, while non-AMF plants grew to only 6

cm. Eastern gamagrass culm height was 61 cm with WL, 55 cm with CL, and 27 cm

with no AMF. Culm height of non-inoculated eastern gamagrass was greater than

non-inoculated big bluestem suggesting slightly better access to P in the absence of

AMF. In contrast, no differences between AMF inoculums was found for tall fescue

as culm height reached 37, 44, and 43 cm with WL, CL, and no AMF, respectively

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(Table 22). Tall fescue culm height with no AMF was not different from big bluestem

with either WL or CL, or eastern gamagrass with CL, but greater than non-inoculated

big bluestem and eastern gamagrass. The tallest plants in this comparison were WL-

inoculated eastern gamagrass averaging 61 cm.

Without AMF, it is likely that either big bluestem or eastern gamagrass would

not survive in habitats where they had to compete against facultative, competitive

species such as tall fescue. Warm-season grasses, including big bluestem, are

dependent on AMF when soil P is limited (Daniels et al., 1988; Brejda et al., 1993;

Noyd et al., 1995; Collins and Foster, 2009). Root growth of tall fescue in this study

was at least 6 times greater than big bluestem with AMF and over 350 times greater

than big bluestem with no AMF (Table 22). Eastern gamagrass with no AMF had 25%

less root growth than tall fescue with or without AMF. However, inoculation with

either WL or CL was equally beneficial to the warm-season grasses, suggesting that

the WL AMF would not be a limiting factor for establishing prairie grasses on the

reclaimed mine soil. In contrast, tall fescue is clearly able to establish quickly in low

P soil, with or without AMF, which demonstrates why it has been widely used in

reclamation of surface mined lands in the eastern U.S. (Roberts et al., 1988a).

Newman and Moser (1988b) found significantly greater adventitious root growth for

tall fescue compared with big bluestem and other warm-season grass during a

glasshouse study comparing root growth at the emergence of a third leaf; however,

third leaf emergence occurred in 15-16 days for big bluestem and 28-32 days for tall

fescue. This suggests that tall fescue seedlings put early resources into root growth.

127

Effect of soil compaction on grass growth

The interaction between grass species and soil compaction was not significant

at α=0.05; however, the response of the each species to increasing soil compaction

was of particular interest in this study. Therefore, the effect of soil compaction on

growth of each species was analyzed with only the WL and CL AMF-inoculated

plants. The non-inoculated plants were not included in the analysis because they

would bias the results. The reclaimed mine soil examined in this research contains

AMF (WL); therefore, a non-AMF scenario is unlikely in the effort to replace the non-

native forage complex with prairie vegetation.

Although tall fescue produced more shoot and root biomass than the other

grasses, increasing compaction did result in a reduction of growth (Figure 5). Tall

fescue shoot biomass was reduced from 6.5 to 4.2 g plant-1

, and root biomass declined

from 4.0 to 2.3 g plant-1

between 1.25 and 1.5 BD, respectively. Furthermore, tall

fescue averaged 22 and 17 tillers plant-1

at 1.0 and 1.25 BD, respectively, and only 14

tillers plant-1

at 1.5 BD; however, no difference was seen in culm height or RSR. Leaf

count also appeared to decline as BD increased, but differences were not detected.

These results conflict with an earlier study that found an increase in plant biomass of

KY-31 tall fescue between 1.6 and 2.0 g cm-3

in loam soil and only a slight decrease in

clay soil (Crews, 1984). However, in a turfgrass study, compaction during summer

resulted in a decrease in root biomass for tall fescue and not for perennial ryegrass

(Lolium perenne L.) (Carrow, 1980), indicating that tall fescue is potentially affected

by compaction.

128

Big bluestem was also affected by compaction as plants at 1.0 BD averaged 18

leaves and 2 tillers, but declined to 11 leaves and 1.2 tillers plant-1

at 1.5 BD (Figure

5). Differences were not found between 1.0 and 1.25 BD. Culm height declined from

63 to 34 cm between 1.25 and 1.5 BD. Shoot and root biomass also declined with the

highest compaction level. Shoot biomass declined from 1.5 and 0.7 g plant-1

and root

biomass declined from 0.7 to 0.3 g plant-1

, between 1.25 and 1.5 BD, respectively.

The RSR did not differ between compaction levels averaging 0.6 at 1.0 BD, and 0.7 at

both 1.25 and 1.5 BD. Warm-season prairie grasses are generally slower to establish

than cool-season forage grasses (Newman and Moser, 1988b; Evanylo et al., 2005)

and big bluestem in this study was consistent with this generalization. Had the plants

grown for a longer period, growth likely would have been greater at all levels of

compaction.

Eastern gamagrass also produced fewer leaves and tillers, and less biomass

than tall fescue; however, it was not negatively affected by any level of compaction

(Figure 5). Leaf counts plant-1

averaged 18 to 24, tiller count averaged 3.6 to 4.8, and

culm height averaged 56 to 62 cm plant-1

. Shoot biomass averaged 1.7 to 2.5 g plant-1

,

root biomass averaged 1.0 to 1.4 g plant-1

, and the RSR remained constant at 0.6 for

each level of compaction. Eastern gamagrass appears slower to establish than tall

fescue; however, eastern gamagrass is effective in rooting through compacted soil and

has cellular compartments (aerenchyma) that allow air flow into deep roots (Clark et

al., 1998; Gilker et al., 2002; Krizek et al., 2003). Eastern gamagrass is also

successful planted in hedgerows for erosion and runoff control, suggesting tolerance

129

of flooding (Dewald et al., 1996; Ritchie et al., 2000), and is as productive as

switchgrass (Panicum virgatum L.) in paired plantings (Edwards et al., 2000).

Conclusions

Establishing native prairie grasses on compacted reclaimed mine soil is limited

by adverse soil conditions. Tall fescue has been successful on reclaimed surface-

mined land in the eastern U.S. coal mining region, but is not native to North America

and is a major component of the low-diversity forage complex planted on mine soil.

Tall fescue establishes quickly but is slightly inhibited by compact soil of at least 1.5 g

cm-3

BD. Big bluestem is a dominant tallgrass prairie species that is slow to establish,

but also is slightly affect by compact soil. Eastern gamagrass is native to the region

and tolerates compacted and wet soil, and may be useful in ameliorating mine soil

compaction, which would facilitate establishment of other native species. Eastern

gamagrass is slower to establish than tall fescue, but is affected relatively less by

compaction. The AMF associated with cool-season forage grasses on reclaimed mine

soil is suitable for establishment of warm-season AMF-dependent prairie grasses;

however, tall fescue is not colonized well by AMF from native tallgrass prairie,

suggesting that tallgrass prairie AMF are more host specific than reclaimed mine soil

AMF. Big bluestem and eastern gamagrass are highly dependent on AMF in low-P

mine soil, whereas, the AMF association with tall fescue is completely facultative.

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Table 20. Soil properties of reclaimed mine soil and arbuscular mycorrhizal fungi

(AMF) pot-culture soil used in a 10-week glasshouse study examining the

affects of soil compaction and AMF on the growth of three grass species.

Mine

2

topsoil

(0-20 cm)

AMF3

Soil Parameter1 WL CL

pH 7.3 7.9 7.7

P (mg kg-1

) 12 7 < 1

K (mg kg-1

) 161 41 77

Ca (mg kg-1

) 3768 1262 1345

Mg (mg kg-1

) 321 198 235

1 Soil P analyzed with Bray P1 method; K, Ca, and Mg analyzed with ammonium

acetate extract method by STAR lab, Wooster, OH.

2 Soil collected from reclaimed surface mine land near Cumberland, OH currently

part of the Wilds (The International Center for the Preservation of Wild

Animals, Inc.).

3 Pot-culture soil containing AMF from the Wilds (WL) reclaimed surface mine

and from Claridon (CL) tallgrass prairie remnant near Marion, OH.

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Table 21. Arbuscular mycorrhizal fungi (AMF) root colonization in three

grasses growing in mine soil during a 10-week glasshouse study.

Grass species AMF1 Colonization (%)

2

Tall fescue WL 29.8 c

CL 9.8 d

None 0.0 d

Big bluestem WL 55.9 b

CL 31.7 c

None 0.0 d

Eastern gamagrass WL 76.2 a

CL 73.3 a

None 0.3 d

1 Sources of AMF are the Wilds (WL), a 30-year old reclaimed surface mine

area near Cumberland, OH, and the Claridon (CL) tallgrass prairie

remnant near Marion, OH.

2 Numbers followed by the same letter are not different using Fisher‘s

protected LSD (α=0.05).

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Table 22. Interaction between grass species and arbuscular mycorrhizal fungi (AMF) on growth parameters in sterilized

mine soil. Sources of AMF are the Wilds (WL), a 30-year old reclaimed surface mine area near Cumberland, OH, and

the Claridon (CL) tallgrass prairie remnant near Marion, OH, in a 10-week glasshouse experiment.

Growth parameters

1

Grass species AMF Leaves Tillers Height Shoot Root RSR

(plant-1

) (plant-1

) (cm plant-1

) (g plant-1

) (g plant-1

) (g g-1

)

Tall Fescue WL 77.4 a 19.4 a 36.7 c 5.9 a 3.8 a 0.7 b

CL 66.0 a 16.5 a 44.3 bc 6.1 a 3.6 a 0.6 b

None 59.8 a 27.9 a 43.1 bc 6.0 a 3.6 a 0.6 b

Big bluestem WL 16.9 cd 1.9 c 49.1 bc 1.1 c 0.6 c 0.8 b

CL 13.5 d 1.4 cd 49.1 bc 1.1 c 0.5 c 0.6 b

None 5.5 e 1.0 d 6.3 e 0.01 e 0.01 e 1.2 a

Eastern gamagrass WL 22.1 b 4.3 b 61.2 a 2.2 b 1.2 b 0.6 b

CL 19.6 bc 3.9 b 54.8 ab 2.0 b 1.2 b 0.6 b

None 6.1 e 1.0 d 26.8 d 0.14 d 0.09 d 0.7 b

1 Numbers in each column followed by the same letter are not different using Fisher‘s protected LSD (α=0.05).

132

133

1.00 1.25 1.50

Lea

ves

(p

lant-1

)

0

20

40

60

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100

1.00 1.25 1.50

Sh

oo

t bio

mas

s (g

pla

nt-1

)

0

2

4

6

8

1.00 1.25 1.50

Til

lers

(p

lant-1

)

0369

1215182124

1.00 1.25 1.50

Ro

ot

bio

mas

s (g

pla

nt-1

)0

1

2

3

4

5

Soil bulk density (g cm-3

)

1.00 1.25 1.50

Hei

gh

t cm

(pla

nt-1

)

0

10

20

30

40

50

60

70

Soil bulk density (g cm-3

)

1.00 1.25 1.50

Ro

ot:

sho

ot

(g g

-1)

0.0

0.2

0.4

0.6

0.8

1.0

a

jk

a

a

a

x xx

kj

a

ab

b

x xx

j

jk

j

l

a

xx

a

a

xjk

k

xx

x

a

b

jk

a

b

a

xx

x

jk jk

ns

nsns

j

Figure 5. Relationships between grass species and soil compaction in sterile mine soil

inoculated with arbuscular mycorrhizal fungi in a 10-week glasshouse study.

Species are tall fescue (—●—), big bluestem (– –■– –), and eastern gamagrass

(····▲····). Differences among compaction levels, for each species, are compared

with Fisher‘s protected LSD (α=0.05) and are shown by the following letters for

each species: tall fescue (a,b,c), big bluestem (j,k,l), eastern gamagrass (x,y,z), and

ns=non-significant.

134

Chapter 7: Summary and Conclusions

This research finds prairie grasses native to North America able to establish on

compacted, calcareous reclaimed mine soil with the potential to increase plant

diversity and ecological function on landscapes with habitats compromised by surface

mining. However, species selection must include those that tolerate limitations caused

by soil compaction, including periods of saturation and flooding due to poor

permeability, and restricted rooting leading to drought and limited mineral availability.

Species most successful in establishing employed different mechanisms to survive on

the mine soil. Eastern gamagrass produced thick, coarse crowns and roots that

penetrated the compacted profile. Tall dropseed established a dense stand as its small

seeds were able to find suitable safe sites to germinate. Tall dropseed was also not

dependent on AMF for growth in low P soil, which is unlike other warm-season

prairie grasses. Buffalograss thrived on the bare soil plots as its spreading stolons

quickly produced a dense ground cover. Furthermore, its natural tolerance of drought

maintained its growth during summer between rainfall events when the clayey topsoil

dried and cracked. Cool-season slender wheatgrass germinated quickly and also

established early ground cover; however, it declined after two years and appeared

stressed by either drought or low available soil nutrients. Virginia wildrye established

135

well in the first planting year, but not well in the second, which may have due to

seedling herbivory. Western wheatgrass had low initial germination but spread by

rhizomes into bare-soil areas and appears to be well adapted to the mine soil habitat.

In contrast, sideoats and blue grama established well in their seedling year, but were

unable to persist into year two. This was likely due to intolerance of anaerobic soil

conditions that persist on the mine soil during late winter and spring. Cool-season

species limited to the arid West did not establish in the first year and likely reflected

incompatibility with the general climate.

Tallgrass and mixed prairie dominants big bluestem, Indiangrass, and little

bluestem had low initial establishment success. Seedling establishment rates were low

and not related to number of seeds sown. These grasses are recognized as slow to

establish but limited number of available safe sites on the mine soil clearly diminished

their success. It was unclear why initial germination rates were low, but seedlings

were observed with elevated crowns through summer of the seedling year, and first

year biomass production was low. Poor germination and early growth left open gaps

in plots seeded with these species. Symbiosis with AMF is critical for survival of

these warm-season grasses when soil P is low; however, the mine soil AMF was found

to be equally or more effective than AMF from the Claridon tallgrass prairie remnant.

Therefore, inadequate AMF was not a limiting factor for establishment on the mine

soil.

The success of non-native forages on the mine soil is also due to mechanisms

that overcome the poor soil habitat. Tall fescue is able to root in compacted soil,

136

establishes early, and is not dependent on AMF when soil P is low. Kentucky

bluegrass appears to avoid the compacted soil by producing thick sod at the soil

surface and tolerating drought periods when they occur. Furthermore, Kentucky

bluegrass sod is a barrier for seedling establishment and may function to reduce

abundance of tall fescue on compacted mine soil when older fescue plants weaken and

die. Bird‘s-foot trefoil is a deep-rooting legume that benefits from symbiosis with

nitrogen fixing bacteria and AMF. These species are also tolerant of saturated soil and

are quick to establish, thus producing a competitive ground cover. Establishing native

prairie communities on mine soil that has supported these non-native forages is

dependent on controlling their reestablishment following planting of prairie species.

Large gaps left open by poor establishment and slow growth of species like big

bluestem are open for reinvasion by the forage species. In this research, plots with

poor establishment were quickly re-colonized by Kentucky bluegrass in the second

year after planting when no fallow year preceded the prairie grass planting; however,

when a fallow year preceded planting, bird‘s-foot trefoil seeds broke dormancy and

established a thick cover (Table 23, Appendix B). Furthermore, intensive use of

herbicides was used to control the invasive Canada thistle during the first two years.

Managing the succession and development of prairie species on the mine soil

landscape is critical for the establishment success of a ‗native‘ mine-soil prairie

community. Finding a compliment of early and late establishing species is critical for

long-term stability and functioning of a new, diverse prairie community. Tall

dropseed, buffalograss, and slender wheatgrass establish easily and produce early

137

ground cover. Eastern gamagrass establishes easily and is highly productive and

produces greater vertical structure than the other grasses. Eastern gamagrass also is

less affected by compaction than the other species in this study, including the non-

native forages, and likely would ameliorate the compaction problem in time as its

roots penetrate deeper into the profile. Late-successional prairie species big bluestem

and Indiangrass may become dominant on mine soil, but would easily be overtaken by

the non-native forage grasses during seedling competition. Therefore, a threshold

must be crossed where native prairie species can establish, reproduce, and replace

themselves on the mine soil without the system reverting back to the non-native forage

complex, or in a different direction, i.e. autumn olive thicket. This will result from

selection of a diverse set of species that incorporates multiple mechanisms to deal with

the soil habitat, and provides opportunities for management inputs, e.g. fire,

herbicides, mowing, grazing, to correct deviations from desired prairie development.

This research also proposes questions regarding how to achieve desired

successional goals, not just for calcareous compacted reclaimed mine soil in Ohio, but

for any restoration problem. Ecological restoration can be viewed as ‗managed

succession‘ where at any point during development, a number of outcomes are

possible given events or conditions that may occur. The challenge is to find

opportunities through species selection or habitat manipulation that create possibilities

for management activities that promote development of desired communities that

ultimately increase ecosystem function on damaged or degraded landscapes.

138

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Appendix A: Photographs of Reclaimed Surface Mined Land in Southeast Ohio.

161

Figure 6. Muskingum Mine near Cumberland, OH. Compacted reclaimed land in

foreground persists in non-native forage complex while non-compacted reclaimed

land in background supports forest vegetation. Photo taken early spring, 2004.

162

Figure 7. Contoured compacted mine soil with non-native forage complex and runoff

collecting in low-lying swales. Photo taken early spring, 2004.

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Figure 8. Eastern deciduous forests in southeast Ohio near Cumberland, OH. Photo

taken Autumn, 2005.

164

Figure 9. Second-year growth of grasses in field plots on reclaimed mine soil at the

Wilds. Plot with dense cover in foreground is tall dropseed, while lighter green

grass with wider leaves in middle of plot area is eastern gamagrass. Non-native

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165

Appendix B: Cover Measurements of Existing Mine Soil Vegetation Re-establishing

in Prairie Grass Plots at the Conclusion of the Research.

166

Table 23. Canopy cover estimates of tall fescue (FEAR), Kentucky bluegrass

(POPR), and bird‘s-foot trefoil (LOCO) three years after the 2005 plantings

and two years following the 2006 planting, following re-establishment in native

grass plots.

Species FEAR POPR LOCO TOTAL

---------------(percent canopy cover)

1---------------

----2005 planting, measurements taken 3 years after planting; no fallow year----

Eastern gamagrass 0 a 8 c 0 c 8 b

Tall dropseed 0 a 11 bc 0 c 11 b

Big bluestem 0 a 79 a 0 c 79 a

Indiangrass 1 a 79 a 1 bc 81 a

Little bluestem 19 a 31 c 4 b 54 a

Sideoats grama 4 a 65 ab 18 a 86 a

Blue grama 4 a 40 bc 18 a 61 a

Buffalograss 9 a 64 ab 0 c 74 a

Virginia wildrye 9 a 62 ab 0 c 71 a

Slender wheatgrass 9 a 68 ab 1 bc 78 a

Continued

167

Table 23. Continued.

Species FEAR POPR LOCO TOTAL

---------------(percent canopy cover)---------------

-2006 planting, measurements taken 2 years after planting and one year of fallow-

Eastern gamagrass 0 a 0 c 0 d 0 d

Tall dropseed 0 a 1 bc 9 bc 9 cd

Big bluestem 0 a 10 ab 35 abc 45 ab

Indiangrass 9 a 0 c 15 abc 24 bc

Little bluestem 4 a 5 a 29 abc 38 ab

Sideoats grama 1 a 1 bc 29 abc 30 bc

Blue grama 1 a 0 c 44 ab 44 ab

Buffalograss 0 a 0 c 4 cd 4 d

Virginia wildrye 0 a 0 c 63 a 63 a

Slender wheatgrass 0 a 0 c 13 cd 13 cd

1 Numbers with the same letter in each column, for each, planting date, are not

statistically different (α=0.05).

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