Development of two sequence-specific PCR markers linked to the le gene that reduces pod shattering in narrow-leafed Lupin (Lupinus angustifolius L.)

Abstract

Wild types of narrow-leaf lupin (Lupinus angustifolius L.) have seed pods that shatter upon maturity, leading to the loss of their seeds before or during the harvest process. Two recessive genes have been incorporated into domesticated cultivars of this species to maximize harvest-ability of the produce. One of these genes is called lentus (le). Two microsatellite -anchored fragment length polymorphism (MFLP) candidate markers were identified as closely linked to the le gene in a recombinant inbred line (RIL) population derived from a domesticated x wild type cross. The candidate MFLP markers were isolated from the gel, re-amplified by PCR, cloned and sequenced. The MFLP polymorphisms were converted into sequence-specific PCR-based markers. Linkage analysis by MapManager indicated that one of the markers, LeM1, was 2.6 centiMorgans (cM) and the other, LeM2, was 1.3 cM from the gene, with both being on the same side. The correlation between the marker genotype and the plant phenotype for the le gene is 95% for the Australian cultivars, and approximately 36% on wild types tested. These markers may be useful in marker assisted selection for the le gene when introgressing wild material into lupin breeding programs.

Grain Legumes

“…This modification results in a reduction of torsional forces upon drying and causes reduced pod shatter (Gladstones 1967). Three simple PCR-based molecular markers closely linked to lentus gene have been developed for narrow-leafed lupin (Boersma et al 2007c;Li et al 2012b). Two of them are dominant markers (LeM1 and LeM2), unable to discriminate between homozygous and heterozygous genotypes (Boersma et al 2007c).…”

“…Three simple PCR-based molecular markers closely linked to lentus gene have been developed for narrow-leafed lupin (Boersma et al 2007c;Li et al 2012b). Two of them are dominant markers (LeM1 and LeM2), unable to discriminate between homozygous and heterozygous genotypes (Boersma et al 2007c). Recently, Li et al (2012b) reported the development of a new codominant marker (LeLi) flanking the lentus gene.…”

Comparative genomics of Lupinus angustifolius gene-rich regions: BAC library exploration, genetic mapping and cytogenetics

“…Examples are mollis, which confers soft seediness (marker MoLi) [5]; tardus and le , which reduce pod shattering (markers TaLi, LeM1, and LeM2) [6,7]; and Ku , which removes the vernalization requirement (marker KuHM1) [8]. Furthermore, the MFLP method was used to develop sets of markers tagging hypothetical genes involved in lupin resistance to pathogenic fungi, including Lanr1, conferring anthracnose resistance (markers AntjM1 and AntjM2) [9-11]; Phr1 and Phr2 , carrying resistance to Phomopsis stem blight, a lupin disease caused by the necrotrophic fungus Diaporthe toxica (markers Ph258M1, Ph258M2, PhtjM1, and PhtjM2) [12,13]; Hua’an Yang, unpublished; and a pair of markers, RustM1 and RustM2, linked to rust-resistance loci [14].…”

Development of a co‐dominant DNA marker linked to the gene lentus conferring reduced pod shattering for marker‐assisted selection in narrow‐leafed lupin (Lupinus angustifolius) breeding

“…Recently, two PCR‐based markers for detecting the presence or absence of lentus gene were developed for narrow‐leafed lupin (Boersma et al. 2007).…”

“…Plant materials: An F 8 recombinant inbred line (RIL) population was developed from a cross between a domesticated breeding line 83A:476 ( lentus/lentus ) and a wild‐type P27255 ( Lentus/Lentus ) of L. angustifolius using a single‐seed‐descent method (Boersma et al. 2007).…”