Estimation of pea (<Emphasis Type="Italic">Pisum sativum</Emphasis> L.) microsatellite mutation rate based on pedigree and single-seed descent analyses |
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Authors: | Jaroslava Cieslarová Pavel Haná?ek Eva Fialová Miroslav Hýbl Petr Smýkal |
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Institution: | 1.Department of Plant Biology,Mendel University,Brno,Czech Republic;2.Department of Grain Legumes,AGRITEC Plant Research Ltd.,?umperk,Czech Republic;3.Department of Plant Biotechnology,AGRITEC Plant Research Ltd.,?umperk,Czech Republic |
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Abstract: | Microsatellites, or simple sequence repeats (SSRs) are widespread class of repetitive DNA sequences, used in population genetics,
genetic diversity and mapping studies. In spite of the SSR utility, the genetic and evolutionary mechanisms are not fully
understood. We have investigated three microsatellite loci with different position in the pea (Pisum sativum L.) genome, the A9 locus residing in LTR region of abundant retrotransposon, AD270 as intergenic and AF016458 located in
5′untranslated region of expressed gene. Comparative analysis of a 35 pair samples from seven pea varieties propagated by
single-seed descent for ten generations, revealed single 4 bp mutation in 10th generation sample at AD270 locus corresponding
to stepwise increase in one additional ATCT repeat unit. The estimated mutation rate was 4.76 × 10-3 per locus per generation, with a 95% confidence interval of 1.2 × 10-4 to 2.7 × 10-2. The comparison of cv. Bohatyr accessions retrieved from different collections, showed intra-, inter-accession variation
and differences in flanking and repeat sequences. Fragment size and sequence alternations were also found in long term in
vitro organogenic culture, established at 1983, indicative of somatic mutation process. The evidence of homoplasy was detected
across of unrelated pea genotypes, which adversaly affects the reliability of diversity estimates not only for diverse germplasm
but also highly bred material. The findings of this study have important implications for Pisum phylogeny studies, variety identification and registration process in pea breeding where mutation rate influences the genetic
diversity and the effective population size estimates. |
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