Morpho-biochemical characterization of a RIL population for seed parameters and identification of candidate genes regulating seed size trait in lentil (Lens culinaris Medik.)

Abstract

The seed size and shape in lentil (Lens culinaris Medik.) are important quality traits as these influences the milled grain yield, cooking time, and market class of the grains. Linkage analysis was done for seed size in a RIL (F5:6) population derived by crossing L830 (20.9 g/1000 seeds) with L4602 (42.13 g/1000 seeds) which consisted of 188 lines (15.0 to 40.5 g/1000 seeds). Parental polymorphism survey using 394 SSRs identified 31 polymorphic primers, which were used for the bulked segregant analysis (BSA). Marker PBALC449 differentiated the parents and small seed size bulk only, whereas large seeded bulk or the individual plants constituting the large-seeded bulk could not be differentiated. Single plant analysis identified only six recombinant and 13 heterozygotes, of 93 small-seeded RILs (<24.0 g/1000 seed). This clearly showed that the small seed size trait is very strongly regulated by the locus near PBLAC449; whereas, large seed size trait seems governed by more than one locus. The PCR amplified products from the PBLAC449 marker (149bp from L4602 and 131bp from L830) were cloned, sequenced and BLAST searched using the lentil reference genome and was found amplified from chromosome 03. Afterward, the nearby region on chromosome 3 was searched, and a few candidate genes like ubiquitin carboxyl-terminal hydrolase, E3 ubiquitin ligase, TIFY-like protein, and hexosyltransferase having a role in seed size determination were identified. Validation study in another RIL mapping population which is differing for seed size, showed a number of SNPs and InDels among these genes when studied using whole genome resequencing (WGRS) approach. Biochemical parameters like cellulose, lignin, and xylose content showed no significant differences between parents and the extreme RILs, at maturity. Various seed morphological traits like area, length, width, compactness, volume, perimeter, etc., when measured using VideometerLab 4.0 showed significant differences for the parents and RILs. The results have ultimately helped in better understanding the region regulating the seed size trait in genomically less explored crops like lentils.

Genome-wide discovery of InDels and validation of PCR-Based InDel markers for earliness in a RIL population and genotypes of lentil (Lens culinaris Medik.)

“…Lentil crop remains susceptible to drought and heat stress, particularly during grain filling stage, which results in forced maturity and reduced yield. With climate change projections indicating increased frequency and intensity of drought and heat stress [ 4 ], there is an urgent need to develop early-maturing lentil cultivars capable of surviving under such challenges with minimum or no yield penalty [ 5 ]. To thrive in cereal-based cropping systems, especially in rice fallow areas in South Asia, there is a demand for extra-early varieties that mature in less than 100 days.…”

Strategies for identifying stable lentil cultivars (Lens culinaris Medik) for combating hidden hunger, malnourishment, and climate variability

“…The grain legume lentil (Lens culinaris Medikus. culinaris) is rich in macro-and micronutrients as well as protein, vitamins, and prebiotic carbohydrates (Taleb et al, 2013;Gupta et al, 2018;Khazaei et al, 2019). Daily consumption of 100 grams of lentils can supply significant amounts of Zn and Fe (Thavarajah et al, 2009;.…”