Marker-assisted pyramiding of lycopene-ε-cyclase, ß-carotene hydroxylase1 and opaque2 genes for development of biofortified maize hybrids.

Malnutrition affects growth and development in humans and causes socio-economic losses. Normal maize is deficient in essential amino acids, lysine and tryptophan; and vitamin-A. Crop biofortification is a sustainable and economical approach to alleviate micronutrient malnutrition. We combined favorable alleles of crtRB1 and lcyE genes into opaque2 (o2)-based four inbreds viz. QLM11, QLM12, QLM13, and QLM14 using marker-assisted backcross breeding. These are parents of quality protein maize versions of two elite hybrids viz. Buland and PMH1, grown in India. Gene-based SSRs for o2 and InDel markers for crtRB1 and lcyE were successfully employed for foreground selection in BC1F1, BC2F1, and BC2F2 generations. The recurrent parent genome recovery ranged from 88.9 to 96.0% among introgressed progenies. Kernels of pyramided lines possessed a high concentration of proA (7.14-9.63 ppm), compared to 1.05 to 1.41 ppm in the recurrent parents, while lysine and tryptophan ranged from 0.28-0.44% and 0.07-0.09%, respectively. The reconstituted hybrids (RBuland and RPMH1) showed significant enhancement of endosperm proA (6.97-9.82 ppm), tryptophan (0.07-0.09%), and lysine (0.29-0.43%), while grain yield was at par with their original versions. The dissemination of reconstituted hybrids holds significant promise to alleviate vitamin-A deficiency and protein-energy malnutrition in developing countries.

Genome wide association mapping for heat tolerance in sub-tropical maize.

BACKGROUND: Heat tolerance is becoming increasingly important where maize is grown under spring season in India which coincide with grain filling stage of crop resulting in tassel blast, reduced pollen viability, pollination failure and barren ears that causes devastating yield losses. So, there is need to identify the genomic regions associated with heat tolerance component traits which could be further employed in maize breeding program. RESULTS: An association mapping panel, consisting of 662 doubled haploid (DH) lines, was evaluated for yield contributing traits under normal and natural heat stress conditions. Genome wide association studies (GWAS) carried out using 187,000 SNPs and 130 SNPs significantly associated for grain yield (GY), days to 50% anthesis (AD), days to 50% silking (SD), anthesis-silking interval (ASI), plant height (PH), ear height (EH) and ear position (EPO) were identified under normal conditions. A total of 46 SNPs strongly associated with GY, ASI, EH and EPO were detected under heat stress conditions. Fifteen of the SNPs was found to have common association with more than one trait such as two SNPs viz. S10_1,905,273 and S10_1,905,274 showed colocalization with GY, PH and EH whereas S10_7,132,845 SNP associated with GY, AD and SD under normal conditions. No such colocalization of SNP markers with multiple traits was observed under heat stress conditions. Haplotypes trend regression analysis revealed 122 and 85 haplotype blocks, out of which, 20 and 6 haplotype blocks were associated with more than one trait under normal and heat stress conditions, respectively. Based on SNP association and haplotype mapping, nine and seven candidate genes were identified respectively, which belongs to different gene models having different biological functions in stress biology. CONCLUSIONS: The present study identified significant SNPs and haplotype blocks associated with yield contributing traits that help in selection of donor lines with favorable alleles for multiple traits. These results provided insights of genetics of heat stress tolerance. The genomic regions detected in the present study need further validation before being applied in the breeding pipelines.

Genetic enhancement of essential amino acids for nutritional enrichment of maize protein quality through marker assisted selection.

Maize grain protein is deficient in two essential amino acids, lysine and tryptophan, defining it as of low nutritive value. The discovery of opaque2 (o2) gene has led to the development of quality protein maize (QPM) that has enhanced levels of essential amino acids over normal maize. However, the adoption of QPM is still very limited. The present study aims at improving the quality of normal four maize inbred lines (LM11, LM12, LM13 and LM14) of single cross hybrids; Buland (LM11 × LM12) and PMH1 (LM13 × LM14) released in India for different agro-climatic zones by introgressing o2 allele along-with modifiers using marker assisted backcross breeding. Both foreground and background selection coupled with phenotypic selection were employed for selection of o2 specific allele and maximum recovery of the recurrent parent genome (87-90%) with minimum linkage drag across the crosses. The converted QPM lines had < 25% opaqueness which is close to the respective recurrent parents. The QPM versions showed high level of tryptophan content ranging from 0.72 to 1.03 across the four crosses. The newly developed best QPM lines were crossed in original combinations to generate QPM hybrids. The grain yield of improved QPM hybrids was at par and there was significant increase in tryptophan content over the original hybrids.The integrated marker assisted, and phenotypic selection approach holds promise to tackle complex genetics of QPM. The dissemination and adoption of improved QPM versions may help to counteract protein-energy malnutrition in developing countries.