Establishment of wild pea Pisum fulvum chromosome segment substitution lines in cultivated P. sativum genetic background, as a tool to study domestication and to broaden genetic diversity.
Petr Smýkal Department of Botany Palacký University in Olomouc, Czech Republic
(*1822 - †1884)
J.G.Mendel and pea diversity exploration
8. February/8. March
1865
Lack of wild accessions in legume germplasm collections
Total numberwild species landraces
breeding
linesvarieties
bean Phaseolus 261 963 2 39 4 10
soya Glycine 229 444 6 17 7 13
groundnut Arachis 128 435 3 31 10 4
chickpea Cicer 98 313 1 50 7 6
pea Pisum 94 001 2 19 8 17
cowpea Vigna 65 323 3 40 4 2
lentil Lens 58 405 3 36 4 3
faba Vicia 43 695 1 32 9 7
pigeonpea Cajanus 40 820 3 49 13 2
lupine Lupinus 38 050 18 27 12 6
bambara Vigna 6 125 1 79 2 1
medic Medicago 91 922 47 6 7 6
clover Trifolium 74 158 53 5 3 6
vetch Vicia 38 460 25 23 3 3
grasspea Lathyrus 26 066 25 21 1 1
trefoil Lotus 8 128 52 6 4 5
1 041 345
Percentage
49248
16910 51450
12396
4980
all wild (2.5%, 1863) 11938
commercial varieties (34%)
breeding lines (13%)
landraces (38%)
mutant stock (2%)
RILs (3.7%)
P. subsp. elatius (0,42%)
"P.humile/syriacum" (1.2%)
"P. transcaucassicum, asiaticum" (0.2%)
P. abyssinicum (0.36%)
P. fulvum (0.46%)
uknown
Pea accessions stratification from 75 324 accessions with known passport data (total 98,000 acc.)
Smýkal P, Coyne C, Redden R, Maxted N (2013) Peas. Chapter 3. In: Singh M, Bisht IS editors, “Genetic and Genomic Resources for Grain Legume Improvement.” Elsevier Insights, London. Pp. 41-80.
Pea germplasm diversity
Jing et al. (2010) BMC Evol. Biol., Smýkal et al. (2012) Plant Genetic Resources
4,538 Pisum accessions held in six European Genebanks
wild Pisum
Pisum genus diversity and phylogeny
Jing et al. (2010) BMC Evol. Biology
Pisum sativum subsp. sativum Pisum fulvum
Pisum sativum subsp. elatius Pisum abyssinicum
Pisum genus geographical distribution variety of habitats and bioclimatic parameters
Secondary genepool Pisum fulvum
P. sativum subsp. abyssinicum
Primary genepool Pisum sativum subsp. sativum subsp. elatius Tertiary genepool
Vavilovia formosa
Pea genepools
Smýkal et al. (2014) Critical Reviews in Plant Sciences
Domestication and selection
Wild species
wild species domestication modern crop
How to reverse the domestication bottleneck
Crop x wild species
Use of crop wild relatives to broad diversity
Jing et al. 2010
P. fulvum WL2140 P. elatius L100
Pea genome – large (4.6 Gbp) and rich in repetitive DNA sequences
9.09 pg DNA/2C haploid genome size of 4.6 Gbp repetitive DNA (70-80%)
Macas et al. (2006) BMC Genomics
Smýkal et al. (2012) Agronomy DPMI
Parent B
Parent A
Principle of introgression lines (backrosses)
I II III IV V VI VII
Linkage group position of used molecular markers
AA212 16
Accox 30
6572
-90
-102
-120
-130
-5
-21
-30
-58
-84
-100
-158
-10
-21
-45
-55
-70
-80
-92
-125
-149
-180
-262
-18-25
-45
-63
-72-79
-120
-180
-19
-30
-55-61
-86
-109
-135
-180
-188
-5
-19
-33
-45
-53
-80
-117
-125
-163
-7
-18
-30
-59
-76
-100
-120
-134
-145
-300
-250
-200
-150
-100
-50
0
0 1 2 3 4 5 6 7 8
Selected 28 microsatellite and 44 gene-specific markers cover all seven linkage groups of pea 4.6 Gbp genome at 2 to 82 cM spacing, with mean of 15.4 cM, delimiting 78 chromosomal regions
Bin-approach
CSSL Finder visualization of genotyped P. fulvum x P. sativum BC3S4 104 lines (unordered)
CSSL Finder selected subset of 49 lines (of 104 analyzed) as sufficiently representative
Issue of nuclear-cytoplasmic conflict
F2 segregation of WL1238 x WL2140 (P.sativum x P. fulvum) 9 albinotic : 31 green
Heritability of mtDNA (cox1) and cpDNA (rbcLA) in F2 plants
cox-1/PsiI rbcLA/AspLEI
WL1238(P.sativum) x WL2140(P.fulvum) WL1238(P.sativum) x WL2140(P.fulvum)
WL1238 x WL2140 WL1238 x WL2140
WL2140 x WL1238
WL2140 x WL1238
WL2140 WL1238
WL2140 x WL1238
WL2140 x WL1238
WL2140 WL1238
Mitochondria Inheritance Chloroplast Inheritance
Selective elimination of cpDNA from P. fulvum P. Smýkal
The heterozygozity was detected by molecular markers in 533 (8%) cases while recurrent Pisum sativum parent in 4552 (69%) and introgressed segments of P. fulvum in 1551 (23%) of 30 cM in average. Taking in account the detection limit achieved with 72 markers for 7LG and spacing total length of 1389 cM of Pisum sativum genome with average spacing 15 cM, the average size of introgressed Pisum fulvum fragmets was estimated to be 40 cM. The were 1.2, 1.3, 2.3, 1.3, 0.8, 1.1 and 1.5 segments per LG. There were 5 to 14 segments per line, with mean of 9.6
Summary of current genotyping
Height (cm) Number of nodes Number of pods
Example of ILs used for gene identification Case of pod dehiscence
Wild pea = pod dehiscent
Cultivated pea = pod indehiscent
• Preliminar phenotyping for 14 traits showing transgression and novel traits.
• Further backcrosses is being undertaken to obtain „single“ fragment lines.
• Upon suffficient homozygozity, lines will be genotyped by medium to high density within LEGATO project partners.
• Such lines will allow phenotypic characterization of unlimited number of target traits, which, coupled together with higher density markers, will provide means for QTL and gene identification and subsequent incorporation in desired genotypes ultimately leading to better performing commercial pea varieties.
• In parallel the series of lines is being established with Pisum sativum subsp. elatius (L100) and P. sativum cv. Cameor parents.
Conclusions and prospects
Kosterin O. Institute of Cytology and Genetics, Novosibirsk, Russia Burstin J., Aubert G. INRA, Dijon, France Gyorgy Kiss AMBIS Ltd., Hungary Peter Winter GenXPro, Germany
Financial support acknowledgement: This work was supported by ME10062 project and current research received funding from the European Community's Seventh Framework Programme (FP7/ 2007-2013) under the grant agreement n°FP7-613551, LEGATO project (2014-2017).
J.G.Mendel W.Bateson Watson/Crick human genom
1865 1900 1902 1915 1942 1953 1970 1973 1983 2000
Sutton-Bovery Maxam/Gilbert/Sanger Morgan Avery
Ch.Darwin McClintock
1944/DNA GMO PCR gene 1859
An International Consortium for Pea Genome Sequencing