Low expression of lipoxygenase 3 (LOX3) enhances the retention of kernel tocopherols in maize during storage.

BACKGROUND: Deficiency of vitamin E results in several neurological and age-related disorders in humans. Utilization of maize mutants with favourable vte4-allele led to the development of several α-tocopherol (vitamin E) rich (16-19 µg/g) maize hybrids worldwide. However, the degradation of tocopherols during post-harvest storage substantially affects the efficacy of these genotypes. METHODS AND RESULTS: We studied the role of lipoxygenase enzyme and Lipoxygenase 3 (LOX3) gene on the degradation of tocopherols at monthly intervals under traditional storage up to six months in two vte4-based contrasting-tocopherol retention maize inbreds viz. HKI323-PVE and HKI193-1-PVE. The analysis revealed significant degradation of tocopherols across storage intervals in both the inbreds. Lower retention of α-tocopherol was noticed in HKI193-1-PVE. HKI323-PVE with the higher retention of α-tocopherol showed lower lipoxygenase activity throughout the storage intervals. LOX3 gene expression was higher (~ 1.5-fold) in HKI193-1-PVE compared to HKI323-PVE across the storage intervals. Both lipoxygenase activity and LOX3 expression peaked at 120 days after storage (DAS) in both genotypes. Further, a similar trend was observed for LOX3 expression and lipoxygenase activity. The α-tocopherol exhibited a significantly negative correlation with lipoxygenase enzyme and expression of LOX3 across the storage intervals. CONCLUSIONS: HKI323-PVE with high tocopherol retention, low -lipoxygenase activity, and -LOX3 gene expression can act as a potential donor in the vitamin E biofortification program. Protein-protein association network analysis also indicated the independent effect of vte4 and LOX genes. This is the first comprehensive report analyzing the expression of the LOX3 gene and deciphering its vital role in the retention of α-tocopherol in biofortified maize varieties under traditional storage.

Promising physiological traits associated with nitrogen use efficiency in rice under reduced N application.

Higher grain yield in high-yielding rice varieties is mostly driven by nitrogen (N) fertilizer applied in abundant amounts leading to increased production cost and environmental pollution. This has fueled the studies on nitrogen use efficiency (NUE) to decrease the N fertilizer application in rice to the possible extent. NUE is a complex physiological trait controlled by multiple genes, but yet to be completely deciphered in rice. With an objective of identifying the promising physiological traits associated with NUE in rice, the performance of 14 rice genotypes was assessed at N0, N50, N100, and N150 for four (two wet and two dry) seasons using agro-morphological, grain yield, flag leaf traits, photosynthetic pigment content, flag leaf gas exchange traits, and chlorophyll fluorescence traits. Furthermore, the data were used to derive various NUE indices to identify the most appropriate indices useful to screen rice genotypes at N50. Results indicate that with the increase in N application, cumulative grain yield increased significantly up to N100 (5.02 t ha-1); however, the increment in grain yield was marginal at N150 (5.09 t ha-1). The mean reduction of grain yield was only 26.66% at N50 ranging from 15.0% to 34.2%. The significant finding of the study is the identification of flag leaf chlorophyll fluorescence traits (Fv/Fm, ΦPSII, ETR, and qP) and Ci associated with grain yield under N50, which can be used to screen N use efficient genotypes in rice under reduced N application. Out of nine NUE indices assessed, NUpE, NUtE, and NUEyield were able to delineate the high-yielding genotypes at N50 and were useful to screen rice under reduced N conditions. Birupa emerged as one of the high yielders under N50, even though it is a moderate yielder at N100 and infers the possibility of cultivating some of the released rice varieties under reduced N inputs. The study indicates the possibility of the existence of promising genetic variability for grain yield under reduced N, the potential of flag leaf chlorophyll fluorescence, and gas exchange traits as physiological markers and best suitable NUE indices to be deployed in rice breeding programs.

Expression Dynamics of lpa1 Gene and Accumulation Pattern of Phytate in Maize Genotypes Possessing opaque2 and crtRB1 Genes at Different Stages of Kernel Development.

Phytic acid (PA) acts as a storehouse for the majority of the mineral phosphorous (P) in maize; ~80% of the total P stored as phytate P is not available to monogastric animals and thereby causes eutrophication. In addition, phytic acid chelates positively charged minerals making them unavailable in the diet. The mutant lpa1-1 allele reduces PA more than the wild-type LPA1 allele. Further, mutant gene opaque2 (o2) enhances lysine and tryptophan and crtRB1 enhances provitamin-A (proA) more than wild-type O2 and CRTRB1 alleles, respectively. So far, the expression pattern of the mutant lpa1-1 allele has not been analysed in maize genotypes rich in lysine, tryptophan and proA. Here, we analysed the expression pattern of wild and mutant alleles of LPA1, O2 and CRTRB1 genes in inbreds with (i) mutant lpa1-1, o2 and crtRB1 alleles, (ii) wild-type LPA1 allele and mutant o2 and crtRB1 alleles and (iii) wild-type LPA1, O2 and CRTRB1 alleles at 15, 30 and 45 days after pollination (DAP). The average reduction of PA/total phosphorous (TP) in lpa1-1 mutant inbreds was 29.30% over wild-type LPA1 allele. The o2 and crtRB1-based inbreds possessed ~two-fold higher amounts of lysine and tryptophan, and four-fold higher amounts of proA compared to wild-type alleles. The transcript levels of lpa1-1, o2 and crtRB1 genes in lpa1-1-based inbreds were significantly lower than their wild-type versions across kernel development. The lpa1-1, o2 and crtRB1 genes reached their highest peak at 15 DAP. The correlation of transcript levels of lpa1-1 was positive for PA/TP (r = 0.980), whereas it was negative with inorganic phosphorous (iP) (r = -0.950). The o2 and crtRB1 transcripts showed negative correlations with lysine (r = -0.887) and tryptophan (r = -0.893), and proA (r = -0.940), respectively. This is the first comprehensive study on lpa1-1 expression in the maize inbreds during different kernel development stages. The information generated here offers great potential for comprehending the dynamics of phytic acid regulation in maize.

Identification of Genomic Regions Associated with High Grain Zn Content in Polished Rice Using Genotyping-by-Sequencing (GBS).

Globally, micronutrient (iron and zinc) enriched rice has been a sustainable and cost-effective solution to overcome malnutrition or hidden hunger. Understanding the genetic basis and identifying the genomic regions for grain zinc (Zn) across diverse genetic backgrounds is an important step to develop biofortified rice varieties. In this case, an RIL population (306 RILs) obtained from a cross between the high-yielding rice variety MTU1010 and the high-zinc rice variety Ranbir Basmati was utilized to pinpoint the genomic region(s) and QTL(s) responsible for grain zinc (Zn) content. A total of 2746 SNP markers spanning a genetic distance of 2445 cM were employed for quantitative trait loci (QTL) analysis, which resulted in the identification of 47 QTLs for mineral (Zn and Fe) and agronomic traits with 3.5-36.0% phenotypic variance explained (PVE) over the seasons. On Chr02, consistent QTLs for grain Zn polished (qZnPR.2.1) and Zn brown (qZnBR.2.2) were identified. On Chr09, two additional reliable QTLs for grain Zn brown (qZnBR.9.1 and qZnBR.9.2) were identified. The major-effect QTLs identified in this study were associated with few key genes related to Zn and Fe transporter activity. The genomic regions, candidate genes, and molecular markers associated with these major QTLs will be useful for genomic-assisted breeding for developing Zn-biofortified varieties.

Identification of alternate dwarfing gene sources to widely used Dee-Gee-Woo-Gen allele of sd1 gene by molecular and biochemical assays in rice ( Oryza sativa L. )

After the success of IR8 and TN1, breeders depended heavily on these two rice cultivars for source of short stature led to the narrow genetic base to majority of present day rice varieties, as far as sd1 (semi-dwarf1) gene is concerned. In addition, analysis of genetic lineage of the majority of the cultivated rice varieties in tropical Asia reveals that sd1 from DGWG (Dee-Gee-Woo-Gen) is the major source of dwarfing gene. Such high amount of genetic homogeneity renders rice plants vulnerable to epidemic of diseases and insect pests. In the current study, we made an attempt to identify the alternate sources of DGWG allele of sd1 gene by characterizing 29 induced and 3 spontaneous dwarf accessions employing marker for DGWG allele of sd1 gene and exogenous application of gibberellic acid (GA3). When occurrence of DGWG allele of sd1 gene and GA3 response were analyzed together, existence of two kinds of dwarfs was noticed viz., dwarf accessions with DGWG allele and dwarf accessions without DGWG allele of sd1 allele exhibiting varying responses to GA3. As many as 22 of 32 dwarf accessions showed absence of DGWG allele of sd1 gene with varying response to GA3 could be used as excellent alternate sources for DGWG allele of sd1 gene. These dwarf accessions could be used for broadening the genetic base for the plant height and thereby minimize the risk of genetic vulnerability. Our strategy of combining molecular and biochemical assays can be efficiently used for identifying alternate dwarfing gene sources to the Green Revolution gene sd1.