Genetic analysis, QTL mapping and gene expression analysis of key visual quality traits affecting the market value of field pea
Visual quality is one of the major factors that determine the market value of field pea (Pisum sativum L.). Breeding for improved visual quality of pea seeds is currently a challenging task, because of the complexity and lack of sound genetic knowledge of the traits. The objectives of this research were to characterize the genetic basis and identify the genomic regions associated with four key visual quality traits (cotyledon bleaching in green pea, greenness in yellow pea, and seed shape and seed dimpling in both green and yellow types) in field pea. Biochemical and gene expression profiling to understand the molecular basis of post-harvest cotyledon bleaching in green pea was also addressed. Two F5:6 recombinant inbred line (RIL) populations (90 lines from Orb X CDC Striker cross, and 120 lines from Alfetta X CDC Bronco cross) were developed and evaluated for visual quality traits in two locations in Saskatchewan, Canada in 2006 and 2007. The four quality traits evaluated all displayed a continuous range of expression with moderate to high heritability. Two genetic linkage maps utilizing 224 markers (29 simple sequence repeat (SSR) (from Agrogene) and 195 amplified fragment length polymorphism (AFLP)) and 223 markers (27 SSR and 196 AFLP ) were constructed for the Orb X CDC Striker population and the Alfetta X CDC Bronco population, respectively. Multiple quantitative traits (QTL) mapping detected major QTLs on linkage group (LG) IV and LG V, as well as location- and year-specific QTLs on LG II and LG III associated with green cotyledon bleaching resistance. Nine QTLs controlling yellow seed lightness, three for yellow seed greenness, 15 for seed shape and nine for seed dimpling were detected. Among them, 5 QTLs located on LG II, LG IV and LG VII were consistent in at least two environments. The QTLs and their associated markers will be useful tools to assist pea breeding programs attempting to pyramid positive alleles for the traits. The bleaching resistant cultivar CDC Striker had a slower rate of chlorophyll degradation in cotyledons and a higher carotenoid to chlorophyll ratio in seed coats than the bleaching susceptible cultivar Orb when seed samples were exposed to high intensity light. An oligo-nucleotide microarray (Ps6kOLI1) was utilized to investigate the gene expression profiles of CDC Striker and Orb seed coats at different developmental stages. It clearly indicated that the expression of genes involved in the production and accumulation of secondary metabolites was significantly different between these cultivars. The results of both biochemical and gene expression studies suggested the bleaching resistance in CDC Striker was not due to the accumulation of chlorophyll pigments in the cotyledons, but rather due to the ability of seed coats to protect them from photooxidation. Accumulation of specific carotenoids which could bind with the reaction center protein complex more effectively and accumulation of phenolic secondary metabolites which could enhance the antioxidant properties and structural integrity of the seed coats may lead to the bleaching resistant phenotype. Therefore, breeding green pea cultivars with higher seed coat antioxidant properties would improve both visual and nutritional quality. This research has provided several insights into molecular approaches to improve field pea visual quality for food markets.
visual quality, Field pea, SSR, AFLP, linkage mapping, QTL analysis, gene expression
Doctor of Philosophy (Ph.D.)