Enhancement of nutritional quality in maize kernel through marker-assisted breeding for vte4, crtRB1, and opaque2 genes.

Traditional maize is poor in vitamin-E [α-tocopherol (α-T): 6-8 ppm], vitamin-A [provitamin-A (proA): 1-2ppm], lysine (0.150-0.2-50%), and tryptophan (0.030-0.040%). Here, we combined favourable alleles of vte4, crtRB1, and opaque2 (o2) genes in the parents of maize hybrids, viz., APQH-10 (PMI-PV-9 × PMI-PV-14) and APQH-11 (PMI-PV-9 × PMI-PV-15) using molecular breeding. Gene-specific markers were successfully used to select vte4, crtRB1, and o2 in BC1F1, BC2F1, and BC2F2 generations. Simple sequence repeats (104-109) were used for background selection, leading to an average recovery of 94% recurrent parent genome. The introgressed inbreds possessed significantly higher α-T: 18.38 ppm, α-/γ-tocopherol (α-/γ-T: 52%), and α-/total tocopherol (α-/TT: 32%) compared to original inbreds (α-T: 8.17 ppm, α-/γ-T: 25%, α-/TT: 18%). These newly derived inbreds also possessed higher ß-carotene (BC: 8.91 ppm), ß-cryptoxanthin (BCX: 1.27 ppm), proA (9.54 ppm), lysine (0.348%), and tryptophan (0.082%) compared to traditional maize inbreds. The reconstituted hybrids recorded higher α-T (2.1-fold), α-/γ-T (1.9-fold), and α-/TT (1.6-fold) over the original hybrids. These reconstituted hybrids were also rich in BC (5.7-fold), BCX (3.3-fold), proA (5.3-fold), lysine (1.9-fold), and tryptophan (2.0-fold) over the traditional hybrids. The reconstituted hybrids had similar grain yield and phenotypic characteristics to original versions. These multinutrient-rich maize hybrids hold great potential to alleviate malnutrition in sustainable and cost-effective manner.

Development and validation of breeder-friendly gene-based markers for lpa1-1 and lpa2-1 genes conferring low phytic acid in maize kernel.

Based on C (wild) to T (mutant) transition at amino acid position 1432 bp of lpa1-1 gene, two dominant markers each specific to wild type (LPA1) and mutant (lpa1-1) allele were developed and validated across seven F2 populations. Joint segregation of these markers behaved in co-dominant fashion, clearly distinguishing heterozygote from two other homozygote genotypes. Full length sequence alignment between wild type (LPA2) and mutant (lpa2-1) allele revealed one transition mutation (A to G) and a co-dominant CAPS marker was developed which differentiated all three types of segregants across seven F2 populations. Across populations, segregants with lpa1-1/lpa1-1 (1.77 mg/g) and lpa2-1/lpa2-1 (1.85 mg/g) possessed significantly lower phytic acid compared to LPA1/LPA1 (2.58 mg/g) and LPA2/LPA2 (2.53 mg/g). Inorganic phosphorus was however higher in recessive homozygotes (lpa1-1/lpa1-1: 0.77 mg/g, lpa2-1/lpa2-1: 0.53 mg/g) than the dominant homozygotes (LPA1/LPA1: 0.33 mg/g, LPA2/LPA2: 0.19 mg/g). Overall, homozygous segregants of lpa1-1 and lpa2-1 showed 31% and 27% reduction of phytic acid, respectively. Analysis of phytate and inorganic phosphorous in the maize kernel in these segregating populations confirmed co-segregation of trait and markers specific to lpa1-1 and lpa2-1. This is the first report of the development of breeder-friendly gene-based markers for lpa1-1 and lpa2-1; and it holds great significance for maize biofortification.

Molecular analysis of mutant granule-bound starch synthase-I (waxy1) gene in diverse waxy maize inbreds.

Waxy corn is popular beacuse of its high amylopectin due to mutation in granule-bound starch synthase-I or Waxy1 (Wx1) gene. Here, we characterized the wx1 allele among 24 diverse waxy inbreds using gene-based markers. A total of 29 alleles with average of 1.81 alleles/locus were observed. Major allele frequency varied from 0.42 to 1.00, with mean of 0.74. The polymorphism information content ranged from 0.00 to 0.56 (average 0.24). Three simple sequence repeat markers, viz., phi027, phi022 and phi061 were more polymorphic in the study. The mean heterozygosity was 0.04, which indicated attainment of higher levels of homozygosity. Dissimilarity coefficient varied from 0.00 to 0.90 with average of 0.51. Seventeen diverse haplotypes of wx1 allele were observed that was consistent with the pedigree. Cluster analyses grouped 24 genotypes into two main clusters each having sub-clusters. The information generated here possesses great potential for improvement of high amylopectin in maize through marker-assisted selection. This is the first report of molecular dissection of wx1 gene among the novel waxy inbreds developed in India.

Gastroprotective effect of the iridoid fraction from Barleria prionitis leaves on experimentally-induced gastric ulceration.

AIM: To study the gastroprotective effect and in vivo antioxidant potential of a standardized iridoid fraction from B. prionitis leaves (BPE) against different gastric ulcer models in rats. METHOD: The standardized iridoid fraction from BPE at 50, 100, and 200 mg/kg body weight was administered orally, twice daily for 5 days for prevention from aspirin, ethanol, cold-restraint stress (CRS), and pylorus ligation (PL)-induced ulcers. Estimation of the antioxidant enzyme activity was carried out in a CRS-induced ulcer model, and various gastric secretion parameters including volume of gastric juice, acid output, and pH value were estimated in the PL-induced ulcer model. RESULTS: BPE showed a dose-dependent ulcer protective effect in PL (18.67%-66.26% protection), aspirin (24.65%-63.25% protection), CRS (20.77%-59.42% protection), and EtOH (16.93%-77.04% protection)-induced ulcers. BPE treatment in PL-rats showed a decrease in acid-pepsin secretion, and enhanced mucin and mucosal glycoproteins. However, BPE reduced the ulcer index with significant decrease in LPO (P < 0.01-0.001), SOD (P < 0.01-0.001), and an increase in CAT (P < 0.01-0.001), activity in the CRS-induced model. CONCLUSION: The data shows that the iridoid fraction from BPE possesses anti-ulcerogenic and antioxidant potential.

Gastroprotective effect of standardized leaf extract from Argyreia speciosa on experimental gastric ulcers in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Argyreia speciosa (L.f), Sweet (Family Convolvulaceae) is used traditionally in Indian System of Medicine as aphrodisiac, rejuvenating agent, intellect promoting agent, brain tonic and in the therapy of hepatomegaly, diabetes and chronic ulcer. AIM OF THE STUDY: To study the gastroprotective effect of standardized butanol fraction of Argyreia speciosa leaf (ASE). MATERIALS AND METHODS: The butanol fraction of Argyreia speciosa leaf (ASE; 50, 100 and 200mg/kg body weight) was administered orally, twice daily for 5 days for prevention from Aspirin (ASP)-, ethanol (EtOH)-, cold-restraint stress (CRS) - and pylorus ligation (PL)-induced ulcers. Estimation of antioxidant enzymes activity was carried out in CRS-induced ulcer model, and various gastric secretion parameters like volume of gastric juice, acid output, and pH value were estimated in PL-induced ulcer model. RESULT: ASE showed dose-dependent ulcer protective effect in ASP 23.64-58.76% (p<0.01 to p<0.001), EtOH 15.45-58.45% (p<0.001), CRS 19.39-78.36% (p<0.001) and PL 19.67-69.04% (p<0.05 to p<0.01), respectively. The percentage of protection by standard drug ranitidine was 77.77-84.32% (p<0.01 to p<0.001) in various gastric ulcer models. The gastric wall mucus was significantly (p<0.001) enhanced by ASE and is regarded as the first line of defence against EtOH-induced gastric ulcers showing cytoprotective property. ASE showed a marginal decrease in volume, acid pepsin concentration and acid pepsin output. However, ASE reduced the ulcer index with significant decrease in LPO level (p<0.001), and SOD level (p<0.01 to p<0.001) as compared with CRS-induced group. A gradual and significant increase in CAT values were observed at 100 and 200mg/kg dose levels (p<0.01 to p<0.001). CONCLUSIONS: The results of our study revealed that Argyreia speciosa possess significant dose dependent gastroprotective activity, probably due to its free radical scavenging activity.