The use of mitochondrial DNA polymorphisms to distinguish commercial cultivars of the broad bean, Vicia faba L.

Abstract

Restriction fragment length polymorphisms (RFLPs) have been used to detect mitochondrial DNA (mtDNA) variation among 9 commercial cultivars of Vicia faba L. The mitochondrial DNA was initially digested with 8 restriction endonucleases revealing complex banding patterns on ethidium bromide (EtBr) stained gels . However, no RFLPs were visualised from these complex profiles . Southern hybridisation using total digested mtDNA as a probe against mtDNA digested with the same restriction enzymes revealed a limited number of RFLPs which allowed the 9 cultivars to be consistently distinguished into two main cytoplasmic types. Southern hybridisation with 23 random mitochondrial clones covering 56 kb of the mitochondrial genome revealed considerable levels of polymorphisms. Of the 23 clones analysed, 12 detected at least 22 polymorphisms using 3 restriction enzymes among the cultivars analysed . These RFLPs allowed the 9 commercial cultivars analysed in this study to be distinguished into at least 6 separate groups. Most polymorphisms distinguished the cultivars into two main cytoplasmic groups .

Molecular tools for utilization of mitochondrial diversity in faba bean (Vicia faba)

“…Finally, we support previous reports on intraspecific variability at the mtDNA level in V. faba (SCALLAN and HARMEY, 1996) because variability in length was detected at nad7 intron 4/5, while variability in primary nucleotide sequence was found within the 5'-end of the second intron of the nad1 gene based on analyses in 16 genotypes of V. faba representative of the worldwide gene pool of this species. The latter locus is one of the most commonly employed non-coding mtDNA regions in various studies in higher plants (e.g.…”

“…Given the lack of cpDNA diversity in V. faba (SHIRAN and MASHAYEKH, 2004;HAIDER et al, 2012) and high mtDNA diversity obtained via PCR-RFLP method and random mtDNA clones used as probes for hybridisation (SCALLAN and HARMEY, 1996) as well as nucleotide and length variability at intron 2/3 of the first gene of the NADH dehydrogenase (nad1 intron 2/3) in the genus Vicia (RYZHOVA et al, 2012), it appears that mtDNA may be a promising tool not only for phylogeographic studies in V. faba that may shed more light on species origin and domestication but also for phylogenetic surveys that may resolve species evolutionary unfolding. However, utilization of maternally inherited (MOGENSEN, 1996) and slowly evolving mtDNA genome (WOLFE et al, 1987;LAROCHE et al, 1997) in diverse studies in V. faba may not be straightforward because plant mtDNA genomes are exceptionally variable in size (200 to 2.900 kb) and organization frequently altered by insertions, deletions and structural re-arrangements (LAROCHE et al, 1997; see also NEGRUK, 2013 and references therein).…”