Influence of rare alleles of β‐carotene hydroxylase and lycopene epsilon cyclase genes on accumulation of provitamin A carotenoids in maize kernels

Vitamin A deficiency is one of the major health problems worldwide. Traditional yellow maize possesses very low provitamin A (proA) concentration in endosperm. The influence of rare alleles of β‐carotene hydroxylase (crtRB1) and lycopene epsilon cyclase (lcyE) genes capable of enhancing proA concentration was studied in four BC₂F₂ populations generated using subtropical inbreds and CIMMYT‐HarvestPlus lines. The occurrence of severe segregation distortion for crtRB1 gene was observed, while lcyE gene was segregated as per Mendelian ratio. Genotype with favourable allele of crtRB1 (CC) had a significant effect on β‐carotene (BC) (7.9‐fold), β‐cryptoxanthin (BCX) (twofold) and proA (5.5‐fold) accumulation, compared to unfavourable genotype (C⁺C⁺). Genotype with favourable allele of lcyE (LL) showed 2.1‐fold, 1.6‐fold and twofold significant enhancement in BC, BCX and proA, respectively, over unfavourable genotype (L⁺L⁺) in pooled analysis. Of the nine genotypes, double homozygote (CC/LL) had the highest mean BC (12.60 μg/g), BCX (4.44 μg/g) and proA (14.82 μg/g), and combined effect was significantly better than individual gene effects or any other combinations. The information generated here would be useful in designing strategy for proA enrichment in subtropical maize.     

Accelerated development of quality protein maize hybrid through marker‐assisted introgression of opaque‐2 allele

Vivek Maize Hybrid 9‐ a popular single‐cross hybrid developed by crossing CM 212 and CM 145 was released for commercial cultivation in India. The parental lines, being deficient in lysine and tryptophan, were selected for introgression of opaque‐2 allele using CML 180 and CML 170 as donor lines through marker‐assisted backcross breeding. The opaque‐2 homozygous recessive genotypes with >90% recovery of the recurrent parent genome were selected in BC₂F_2, and the seeds with <25% opaqueness in BC₂F₃ were forwarded for seed multiplication. Vivek Quality Protein Maize (QPM) 9, the improved QPM hybrid, showed 41% increase in tryptophan and 30% increase in lysine over the original hybrid. The grain yield of the improved hybrid was on par with the original hybrid. The newly improved QPM maize hybrid released in 2008 will help in reducing the protein malnutrition because its biological value is superior over the normal maize hybrids. This short duration QPM maize hybrid has been adopted in several hill states of North Western and North Eastern Himalayan regions.     

Marker‐assisted introgression of lpa2 locus responsible for low‐phytic acid trait into an elite tropical maize inbred (Zea mays L.)

Maize is an important food and feed crop worldwide. Phytic acid (PA), in maize kernel, is an antinutritional factor. PA chelates mineral cations and causes mineral deficiency in humans and phosphorous deficiency in animals. The undigested PA excreted by monogastric animals causes phosphorous eutrophication. Therefore, development of low‐phytate maize is indispensable. The low‐phytate locus (lpa2 allele) has been transferred from low‐phytate mutant line ‘EC 659418’ into an elite inbred UMI 395 through marker‐assisted backcross breeding (MABB). The MABB involved three backcrosses followed by two selfing steps, including ‘foreground selection’, that is, selecting lines with lpa2 allele with the help of a codominant SSR marker ‘umc2230’ and ‘background selection’, that is, selecting plants having genetic background similar to that of the recurrent parent using 50 codominant SSR markers. Two low‐phytate lpa2 lines with genome similar (>90% similarity) to that of recurrent parent have been identified. These lines can be used as parent in future hybridization programmes for obtaining low‐phytate high‐yielding maize hybrids.     

Marker‐assisted backcrossing of a null allele of the α‐subunit of Soybean (Glycine max) β‐conglycinin with a Chinese soybean cultivar (a). The development of improved lines

The development of soybean varieties that lack the β‐conglycinin α‐subunit is an attractive goal because the β‐conglycinin α‐subunit negatively influences the nutrition and gelation of tofu and is a major allergen. To remove this undesirable allergen and simultaneously improve the seed nutritional value and food‐processing quality, marker‐assisted background selection (MABS) was used in backcross breeding to incorporate cgy‐2, a null phenotype version of the gene encoding the β‐conglycinin α‐subunit, from the donor line ‘RiB’ into the genetic background of the Chinese cultivar ‘Dongnong47’ (DN47), a popular high‐oil superfine seed soybean cultivar from Heilongjiang Province, China. In each F₂ (F₂, BC_nF₂) generation of the breeding programme, the offspring that carried the introgressed cgy‐2 were identified by sodium dodecyl sulphate–polyacrylamide gel electrophoresis and rescreened by MABS using simple sequence repeat markers to accelerate recurrent parent genome recovery. Of the 49 advanced backcrossing breeding lines (ABLs), the three best lines, ABL1, ABL2 and ABL3, were selected from the BC₁, BC₂ and BC₃ populations, respectively. The ABLs were evaluated for desirable agronomic characteristics, yield‐related traits, amino acid composition, free amino acid composition and tofu‐processing quality in the mature seeds. All of the ABLs lacked the α‐subunit but grew and reproduced normally without deleterious effects on physiological processes such as seed development and germination. The free amino acid content of ABL1 was significantly higher than that of ‘DN47’, with arginine (Arg) being particularly enriched. Compared to the recurrent parent ‘DN47’, the total protein content of the three ABLs was higher, the amino acid composition of the seed proteins was markedly modified and the yield and hardness of the tofu that was made from the ABLs were significantly increased. MABS combined with stringent phenotypic selection in a backcross breeding programme is a feasible strategy for the genetic engineering of seed protein components to produce allergenic subunit‐deficient variant alleles.     

Effects of the Gpc‐B1 locus on high grain protein content introgressed into Argentinean wheat germplasm

Wheat grain protein content (GPC) is important for human nutrition and has a strong influence on the quality of pasta and bread. The objective of this study was to analyse the introduction of the Gpc‐B1 allele into two Argentinean bread wheat cultivars. Near‐isogenic lines were developed in ‘ProINTA Oasis’ and ‘ProINTA Granar’ using marker‐assisted selection. Gpc‐B1 lines showed a significant (P = 0.01) increase in GPC and a significant (P = 0.001) decrease in grain weight in comparison with control lines without Gpc‐B1. Differences in yield were not significant (P = 0.49) between lines. Gpc‐B1 lines significantly reduced (P = 0.02) straw nitrogen concentration at maturity and significantly increased (P = 0.02) the nitrogen harvest index. When data were analysed by genotype and environment, differences in some analysed parameters were found, indicating that Gpc‐B1 expression may be affected by different genetic backgrounds and environmental conditions. These results suggest that the introgression of the Gpc‐B1 allele into Argentinean wheat germplasm could be a valuable resource for improving GPC with no detrimental effect on grain yield.