Construction of an intraspecific linkage map of lentil (Lens culinaris ssp. culinaris)

Abstract

The first intraspecific linkage map of the lentil genome was constructed with 114 molecular markers (100 RAPD, 11 ISSR and three RGA) using an F(2) population developed from a cross between lentil cultivars ILL5588 and ILL7537 which differed in resistance for ascochyta blight. Linkage analysis at a LOD score of 4.0 and a maximum recombination fraction of 0.25 revealed nine linkage groups comprising between 6 and 18 markers each. The intraspecific map spanned a total length of 784.1 cM. The markers were distributed throughout the genome, however markers were clustered in the middle or near the middle of the linkage groups, suggesting the location of centromeres. Of 114 mapped markers, 16 (14.0%) were distorted, usually at the end or middle of the linkage groups. The utility of ISSR and RGA markers for mapping in lentil was explored, and the primer with an (AC) repeat motif was found to be useful.

An intersubspecific genetic map of Lens

“…Molecular markers, RAPDs, AFLPs and ISSRs, have already proven their usefulness for Lens genetic mapping (Eujayl et al 1997(Eujayl et al , 1998Rubeena et al 2003), but microsatellite markers have not been previously used in lentil mapping. AFLPs have provided a robust method for producing linkage maps in breeding populations that have a relatively narrow genetic base (Powell et al 1997).…”

“…The previous most extensive lentil maps obtained from inter-subspecific and intraspecific crosses, respectively, cover 1,073 cM with an average distance between markers of 6.0 cM (Eujayl et al 1998), and 784.1 cM with a density of 6.9 cM between adjacent markers (Rubeena et al 2003). As the physical size of the lentil genome was estimated to be 4,086 Mpb (Arumuganathan and Earle 1991), 1 cM in the present map would represent 1.88 Mpb on average, which is half the value of 3.8 Mpb/cM calculated in the inter-subspecific map obtained by Eujayl et al (1998).…”

“…ISSR markers are highly polymorphic and are useful in studies on genetic diversity, phylogeny, gene tagging, genome mapping and evolutionary biology (Reddy et al 2002). They have been used in many crop species, including legumes (Ratnaparkhe et al 1998;Ajibade et al 2000;Bornet and Branchard 2001;Chowdhury et al 2002;Iruela et al 2002;Rajesh et al 2003), and have proved their usefulness in genetic mapping and marker-assisted selection (Zietkiewicz et al 1994;Ratnaparkhe et al 1998;Winter et al 2000;Rubeena et al 2003). ISSR markers have already been used in lentil to evaluate genetic variation in a collection of cultivated lentils (Zµvodnµ et al 2000;Sonante and Pignone 2001).…”

“…A low genetic variability and insufficient genetic information are the reasons why until recently genetic maps of this species consisted of a relatively small number of markers, mainly isozymes and restriction fragment length polymorphisms (RFLPs) that covered an also relatively small portion of the lentil genome (Havey and Muehlbauer 1989;Weeden et al 1992;Tahir et al 1993). The recent development of a large number of molecular markers, especially RAPDs (randomly amplified polymorphic DNA) and AFLPs (amplified fragment length polymorphisms), is now providing the basis for constructing saturated maps in this species (Eujayl et al 1997(Eujayl et al , 1998Rubeena et al 2003), which consequently enables for the localization of genes of agronomic interest and facilitates a faster improvement in breeding programs (Tullu et al 2003). Inter-simple sequence repeats (ISSRs) are a relatively new type of DNA marker that involves the direct use of microsatellite sequences as primers in PCR analyses (Gupta et al 1994) to obtain DNA markers.…”

Application of biotechnology in breeding lentil for resistance to biotic and abiotic stress

“…Restriction fragment length polymorphism (RFLP) markers, developed from cutting genomic DNA with restriction enzymes and separating the resulting DNA fragments with electrophoresis, were the first type of molecular marker used in the construction of a lentil genetic linkage map (Havey & Muehlbauer, 1989). Subsequently, arbitrarily produced polymerase chain reaction (PCR)-based markers, such as random amplified polymorphic DNA (RAPD) were used to study diversity, phylogeny and taxonomy of Lens (Ford et al, 1997;Ferguson et al, 2000;Sharma et al, 1996), to develop linkage maps (Eujayl et al, 1997(Eujayl et al, , 1998aRubeena et al, 2003), to tag genes of interest (Chowdhury et al, 2001;Eujayl et al, 1998bEujayl et al, , 1999Ford et al, 1999;Tullu et al, 2003) and to determine pathogen population structure (Ford et al, 2000). Arbitrarily produced amplified fragment length polymorphism (AFLP) markers have also been used in lentil linkage mapping (Durán et al, 2004;Eujayl et al, 1998a;Hamwieh et al, 2005;Kahraman et al, 2004) and to study genetic diversity (Sharma et al, 1996), differentiate cultivars (Závodná et al, 2000) and identify markers linked to specific traits .…”