Bio-Controls: Bacterial Bio-Controls: Bacterial Infection FundamentalsInfection Fundamentals

By Ryan MurrayBy Ryan Murray

NeedNeed

Crops decimated by Pythium (Stietsma, 1973)

NeedNeed

Corn Fields killed by Stewarts

Former Methods of ControlFormer Methods of Control DDTDDT

Dioxin (Agent Dioxin (Agent Orange)Orange)

Bio ControlsBio Controls Espinoza (2006) and Garsney (2001)Espinoza (2006) and Garsney (2001)

PurposePurpose Finding bio controls- fight off Finding bio controls- fight off

diseases and pestsdiseases and pestsAlternate HypothesesAlternate Hypotheses

1.If Soybean Plants exposed to Pythium and Fusarium are treated with P.Putida, Fungal Mycelium, and S. griesevoridis, the infections will decrease2. If soybean plants exposed to Root Knot Nematodes are treated with Phasmarhabditis hermaphrodita, the infestation will decrease.

Pythium Root RotPythium Root Rot

(Gubler, 1999)(Shapiro, 2002)

Lit ReviewLit Review (Sietsma,1973) Wall structure and (Sietsma,1973) Wall structure and

Growth in Pythium Bacterial Growth in Pythium Bacterial Infection. Cambridge University PressInfection. Cambridge University Press

Fungal Mycelium

FusariumFusarium

(Ajello,1979)

Lit ReviewLit Review (Tari,1988) Fusarium Wilt Suppression and (Tari,1988) Fusarium Wilt Suppression and

Agglutinability of Pseudomonas putida. Agglutinability of Pseudomonas putida. Appl Environ Microbiol. AugustAppl Environ Microbiol. August19881988 Leonard KJ, Bushnell WR. 2003. Fusarium Leonard KJ, Bushnell WR. 2003. Fusarium

head blight of wheat and barley. American head blight of wheat and barley. American Phytopathological Society Press .Phytopathological Society Press .

Root Knot Nematodes Root Knot Nematodes Ester (2003) Ester (2003) in cooperation with the

U.S. Department of Agriculture. Stanley R. Johnson, director, Cooperative Extension Service, Iowa State University of Science and Technology,Ames, Iowa.

Rhabditid Nematode attacking root knots

Root- Knot Nematodes

Stewart’s DiseaseStewart’s Disease

(Munkvold, 2000)

(Stevens-Boewe, 2006)

Pseudomonas PutidaPseudomonas Putida

(Kachlany, 2002)

S. griseoviridisS. griseoviridis Against resident levels of Fusarium-Against resident levels of Fusarium-

reduced Fusarium infection by 16%reduced Fusarium infection by 16%

Similarity Similarity Distinct similarities between the three Distinct similarities between the three

diseases.diseases.SOYBEAN PLANTS SOYBEAN PLANTS

Soybean yields are affected quantitatively and qualitatively by fungi, insects, nematodes, and viruses (Domier, 2009)

METHODOLOGY METHODOLOGY N= 50 Soybean Plants

N=10Exposed to Pythium

N=10Exposed to Fusarium

N=10 Exposed to Stewart’s (rootknots)

Treated with P.putida, Fungal Mycelium and S. griseoviridis

Treated with Phasmarhabditis hermaphrodita

Time = 30 daysMeasurements taken using Diluting Fluid (measuring volume of bacterial infection) and Leaf health scale

SPPS Statistical Analysis by ANOVA comparing the various bio controls to the control followed by Sheffe Post Hoc Test where p<.05

N=10 ControlN=10 Exposed to Root Knots (No Stewarts)

Protocols for BacteriaProtocols for Bacteria All strains must be kept isolated from All strains must be kept isolated from

one another to avoid contaminationone another to avoid contamination Plants infected by injection or pin-Plants infected by injection or pin-

prick methodprick method Root Knots exposed to Stewarts by Root Knots exposed to Stewarts by

ingestion in mediumingestion in medium

Budget PlanBudget PlanVendor

Order Number Quant Description Price

Carolina Online 155265 1 Pseudomonas Putida, Living tube $9.20

Carolina NP-15-6033 1 Fusarium Oxysporum $9.20

Cornell University-Xiaohong Wang 1 Root Knot Nematodes

Carolina Online 593120 1 Walter QZG Zoom Stereomicroscope $519.00

BD CATALOG 442104 1 Diluting fluid- used to count Bacteria

Carolina Online 721169 1 Pyrex® Vista™ Test Tubes 50 count $14.50

Cornell University-Xiaohong Wang 1 Phasmarhabditis hermaphrodita

Carolina Online 158443 1 Bean, Soy, Viable Seed, 1 lb $8.75

Still Searching Fungal Mycelium

Do- AbilityDo- Ability1.1. Pythium, Fusarium, and Root Knots Pythium, Fusarium, and Root Knots

obtainable by online and collegiate obtainable by online and collegiate sourcessources

2.2. P.Putida, Mycelium, P.Putida, Mycelium, S. griseoviridis,S. griseoviridis, and and Phasmarhabditis Phasmarhabditis hermaphrodita; available online at hermaphrodita; available online at Carolina and at other sourcesCarolina and at other sources

BibliographyBibliography Bagnall, R.. Control of Pythium wilt and root rot of hydroponically grown lettuce by means of Bagnall, R.. Control of Pythium wilt and root rot of hydroponically grown lettuce by means of

chemical treatment of the nutrient solution. University of Pretoria Biological Journal. 2007. chemical treatment of the nutrient solution. University of Pretoria Biological Journal. 2007. Pgs 4-24Pgs 4-24

Bergstrom, Gary C. Biocontrol for plants with Bacillus subtilis, Pseudomonas putida, and Bergstrom, Gary C. Biocontrol for plants with Bacillus subtilis, Pseudomonas putida, and Sporobolomyces roseus. Cornell Research Foundation, Inc. 2005.pgs 45-59Sporobolomyces roseus. Cornell Research Foundation, Inc. 2005.pgs 45-59

Gilardi, Gerald. “Pseudomonas”. McGraw-Hill Encyclopedia of Science and Technology. ©1992Gilardi, Gerald. “Pseudomonas”. McGraw-Hill Encyclopedia of Science and Technology. ©1992 Grewal, P.S., Ehlers, R.-U., Shapiro-Ilan, D.I. Nematodes as Biocontrol Agents. CAB Grewal, P.S., Ehlers, R.-U., Shapiro-Ilan, D.I. Nematodes as Biocontrol Agents. CAB

International Publisher (2006) pp. 505 International Publisher (2006) pp. 505 Howard ,DH. Pathogenic Fungi in Humans and Animals (2nd ed.)New York. Routledge, USA. Howard ,DH. Pathogenic Fungi in Humans and Animals (2nd ed.)New York. Routledge, USA.

(2003)(2003) Jarvis, W.R. Managing Diseases in Greenhouse Crops. American Phytopathological Society Jarvis, W.R. Managing Diseases in Greenhouse Crops. American Phytopathological Society

Press. St. Paul. 1992.Pg 288. Press. St. Paul. 1992.Pg 288. Kloepper. Bacterial cultures for root-colonizing plants.Cominco Fertilizers Research. Volume 6. Kloepper. Bacterial cultures for root-colonizing plants.Cominco Fertilizers Research. Volume 6.

April, 1996.Pg 71. April, 1996.Pg 71. Leonard KJ, Bushnell WR. Fusarium head blight of wheat and barley. American Leonard KJ, Bushnell WR. Fusarium head blight of wheat and barley. American

Phytopathological Society Press .(2003) pgs 181-185Phytopathological Society Press .(2003) pgs 181-185 Owen-Going, T.N.; Sutton, J.C.; Grodzinski, B. Relationships of Owen-Going, T.N.; Sutton, J.C.; Grodzinski, B. Relationships of PythiumPythium isolates and sweet isolates and sweet

pepper plants in single-plant hydroponic units. Canadian Journal of Plant Pathology. 2003 pepper plants in single-plant hydroponic units. Canadian Journal of Plant Pathology. 2003 Volume25. Pages 155-167. Volume25. Pages 155-167.

Walsh TJ, Dixon DM “Spectrum of Mycoses”. Baron's Medical Microbiology (4th ed.).(1996).Walsh TJ, Dixon DM “Spectrum of Mycoses”. Baron's Medical Microbiology (4th ed.).(1996). Wyss, P. Testing the effect of biological control agents on the formation of vesicular Wyss, P. Testing the effect of biological control agents on the formation of vesicular

arbuscular mycorrhiza. Plant and Soil Journal. Volume 147, Number 1.November, 1992. 159-arbuscular mycorrhiza. Plant and Soil Journal. Volume 147, Number 1.November, 1992. 159-162162