A novel micronutrients and methyl jasmonate cocktail of elicitors via seed priming improves drought tolerance by mitigating oxidative stress in rice (Oryza sativa L.).

Drought is a major limiting factor for rice (Oryza sativa L.) production globally, and a cost-effective seed priming technique using bio-elicitors has been found to have stress mitigating effects. Till date, mostly phytohormones have been preferred as bio-elicitors, but the present study is a novel attempt to demonstrate the favorable role of micronutrients-phytohormone cocktail, i.e., iron (Fe), zinc (Zn), and methyl jasmonate (MJ) via seed priming method in mitigating the deleterious impacts of drought stress through physio-biochemical and molecular manifestations. The effect of cocktail/priming was studied on the relative water content, chlorophyll a/b and carotenoid contents, proline content, abscisic acid (ABA) content, and on the activities of ascorbate peroxidase (APX), superoxide dismutase (SOD), NADPH oxidase (Nox), and catalase (CAT). The expressions of drought-responsive genes OsZn-SOD, OsFe-SOD, and Nox1 were found to be modulated under drought stress in contrasting rice genotypes -N-22 (Nagina-22, drought-tolerant) and PS-5 (Pusa Sugandh-5, drought-sensitive). A progressive rise in carotenoids (10-19%), ABA (18-50%), proline (60-80%), activities of SOD (27-62%), APX (46-61%), CAT (50-80%), Nox (16-30%), and upregulated (0.9-1.6-fold) expressions of OsZn-SOD, OsFe-SOD, and Nox1 genes were found in the primed plants under drought condition. This cocktail would serve as a potential supplement in modern agricultural practices utilizing seed priming technique to mitigate drought stress-induced oxidative burst in food crops.

Soybean (Glycine max) isoflavone conjugate hydrolysing ß-glucosidase (GmICHG): a promising candidate for soy isoflavone bioavailability enhancement.

Isoflavones are a sub-class of phenylpropanoids having health benefits and a role in plant defence and plant-rhizobium interaction. Isoflavone conjugate hydrolysis is crucial in determining the bioactivity and bioavailability of these isoflavones inside the human body. This study examined the different characteristics of soy isoflavone conjugate hydrolysing ß-glucosidase (GmICHG) to explore its potential for isoflavone bioavailability enhancement. We cloned the full-length GmICHG cDNA from the soybean seedling roots from the DS2706 variety of 1545 bp. The bioinformatics analysis revealed secretion and glycosylation of this protein. The evolutionary relatedness of this gene to the other glucosidases interestingly had related sequences outside the Papilionaceae family. The protein had a pI above neutral of 7.62 and optimum pH of 6.0, indicating its activity in the extracellular acidic environment. The GmICHG gene expression at three stages of seedling roots gradually rose to 1.84 ± 0.54 fold and a concomitant increase in the ß-glucosidase activity. The enzyme kinetics of GmICHG showed a K m of 6.38 mM and V max of 2.82 U/ml and an optimum temperature of 40 °C. These hint that soy ICHG can be a potent candidate for the isoflavone bioavailability enhancement by hydrolysing their ß-glycosidic bonds. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03427-5.

Starch molecular configuration and starch-sugar homeostasis: Key determinants of sweet sensory perception and starch hydrolysis in pearl millet (Pennisetum glaucum).

Starch-sugar homeostasis and starch molecular configuration regulates the dynamics of starch digestibility which result in sweet sensory perception and eliciting glycemic response, which has been measured in vitro as inherent glycemic potential (IGP). The objective of the research was to understand the key determinants of IGP as well as sweetness in different Pearl millet (PM) genotypes. To understand the intricate balance between starch and sugar, total starch content (TSC) and total soluble sugars (TSS) were evaluated. Higher concentrations of TSC (67.8%), TSS (2.75%), glucose (0.78%) and sucrose (1.68%) were found in Jafarabadi Bajra. Considering the role of compact molecular configuration of starch towards digestibility, X-ray powder diffraction (XRD) analysis was performed. A-type crystallinity with crystallinity degree (CD %) ranged from 53.53-62.63% among different genotypes, where the least CD% (53.53%) was found in Jafarabadi Bajra. In vitro starch hydrolyzation kinetics carried out to determine IGP, revealed a maximum of 77.05% IGP with minimum 1.42% resistant starch (RS) in Jafarabadi Bajra. Overall our results suggest higher sweet sensory perception of Jafarabadi Bajra which is contributed by the matrix composition with least molecular compactness of starch. Also, the interdependence among starch quality parameters; CD%, IGP, RS and amylose has also been discussed.

Role of nutraceutical starch and proanthocyanidins of pigmented rice in regulating hyperglycemia: Enzyme inhibition, enhanced glucose uptake and hepatic glucose homeostasis using in vitro model.

Dysregulation of glucose homeostasis result in hyperglycemia and pigmented rice, unique combination of high quality starch and phenolics has the potential in regulating it. In this study, pigmented rice was characterized in terms of nutraceutical starch (NS) and phenolic content. Further the effect of rice phenolics on carbolytic enzyme inhibition, glucose uptake, hepatic glucose homeostasis and anti-glycation ability was analyzed in vitro. The most relevant effect on enzyme inhibition (α-amylase: IC50-42.34 µg/mL; α-glucosidase: IC50:63.89 µg/mL), basal uptake of glucose (>39.5%) and anti-glycation ability (92%) was found in red rice (RR), than black rice (BR). The role of RR phenolics in regulating glucose homeostasis was deciphered using hepatic cell line system, which found up-regulation of glucose transporter 2 (GLUT2) and glycogen synthase 2 (GYS2); while expression of gluconeogenic genes were found down regulated. To our knowledge this study is the first report validating the role of starch-phenolic quality towards anti-hyperglycemic effect of RR.

Protein and gene integration analysis through proteome and transcriptome brings new insight into salt stress tolerance in pigeonpea (Cajanus cajan L.).

Salt stress is a major constrain to the productivity of nutritionally rich pigeonpea, an important legume of SE Asia and other parts of the world. The present study provides a comprehensive insight on integrated proteomic and transcriptomic analysis of root and shoot tissues of contrasting pigeonpea varieties (ICP1071- salt-sensitive; ICP7- salt-tolerant) to unravel salt stress induced pathways. Proteome analysis revealed 82 differentially expressed proteins (DEPs) with ≥±1.5 fold expression on 2-Dimensional (2D) gel. Of these, 25 DEPs identified through MALDI-TOF/TOF were classified using Uniprot software into functional categories. Pathways analyses using KAAS server showed the highest abundance of functional genes regulating metabolisms of carbohydrate followed by protein folding/degradation, amino acids and lipids. Expression studies on six genes (triosephosphate isomerase, oxygen evolving enhancer protein 1, phosphoribulokinase, cysteine synthase, oxygen evolving enhancer protein 2 and early nodulin like protein 2) with ≥±3 fold change were performed, and five of these showed consistency in transcript and protein expressions. Transcript analysis of root and shoot led to positive identification of 25 differentially expressed salt-responsive genes, with seven genes having ≥±5 fold change have diverse biological functions. Our combinatorial analysis suggests important role of these genes/proteins in providing salt tolerance in pigeonpea.

Pullulanase activity: A novel indicator of inherent resistant starch in rice (Oryza sativa. L).

Starch quality studies over the decades highlighted the immense health benefits of resistant starch (RS), but still much is needed to elucidate the science behind its inherent formation. Till date, there is no report to establish the relationship between inherent RS content and pullulanase (PUL) activity in any of the crops. In this study, we emphasize the novel role of debranching enzyme, PUL towards inherent RS formation, using rice as a model crop. Biochemical analysis of 51 rice genotypes for amylose content (AC) revealed a good amount of variation ranging from 7.03 to 37.33%. Further, hierarchical clustering which resulted in 11 genotypes of varying RS (0.33-2.7%), highlighted medium dependency towards amylose and low dependency towards amylopectin content. The discrete differences in microstructure, unimodal distribution and tight packing of starch granules observed in higher RS genotype indicated the higher possibility of compact cluster structure of amylopectin, modulated by PUL. Qualitative and quantitative assays performed validated the relevant role of PUL towards inherent RS content with very high dependency score (R2 = 0.98). This is the first report regarding the fact that higher PUL activity contribute to inherent RS using novel hypothetical 'Pullulanase-Amylopectin Trimming Model'.

Physiological, Biochemical, Epigenetic and Molecular Analyses of Wheat (Triticum aestivum) Genotypes with Contrasting Salt Tolerance.

Abiotic stress exerts significant impact on plant's growth, development, and productivity. Productivity of crop plants under salt stress is lagging behind because of our limited knowledge about physiological, biochemical, epigenetic, and molecular mechanisms of salt tolerance in plants. This study aimed to investigate physio-biochemical, molecular indices and defense responses of selected wheat cultivars to identify the most contrasting salt-responsive genotypes and the mechanisms associated with their differential responses. Physio-biochemical traits specifically membrane stability index, antioxidant potential, osmoprotectants and chlorophyll contents, measured at vegetative stage, were used for multivariate analysis to identify the most contrasting genotypes. Genetic and epigenetic analyses indicated the possible mechanisms associated with differential response of the wheat genotypes under salt stress. Better antioxidant potential, membrane stability, increased accumulation of osmolytes/phytophenolics, and higher K+/Na+ ratio under 200 mM NaCl stress identified Kharchia-65 to be the most salt-tolerant cultivar. By contrast, increased MDA level, reduced soluble sugar, proline, total chlorophyll, total phenolics contents, and lower antioxidant potential in HD-2329 marked it to be sensitive to the stress. Genetic and bioinformatics analyses of HKT1;4 of contrasting genotypes (Kharchia-65 and HD-2329) revealed deletions, transitions, and transversions resulting into altered structure, loss of conserved motifs (Ser-Gly-Gly-Gly and Gly-Arg) and function in salt-sensitive (HD-2329) genotype. Expression analysis of HKTs rationalized the observed responses. Epigenetic variations in cytosine methylation explained tissue- and genotype-specific differential expression of HKT2;1 and HKT2;3.

Salt-Induced Tissue-Specific Cytosine Methylation Downregulates Expression of HKT Genes in Contrasting Wheat (Triticum aestivum L.) Genotypes.

Plants have evolved several strategies, including regulation of genes through epigenetic modifications, to cope with environmental stresses. DNA methylation is dynamically regulated through the methylation and demethylation of cytosine in response to environmental perturbations. High-affinity potassium transporters (HKTs) have accounted for the homeostasis of sodium and potassium ions in plants under salt stress. Wheat (Triticum aestivum L.) is sensitive to soil salinity, which impedes its growth and development, resulting in decreased productivity. The differential expression of HKTs has been reported to confer tolerance to salt stress in plants. In this study, we investigated variations in cytosine methylation and their effects on the expression of HKT genes in contrasting wheat genotypes under salt stress. We observed a genotype- and tissue-specific increase in cytosine methylation induced by NaCl stress that downregulated the expression of TaHKT2;1 and TaHKT2;3 in the shoot and root tissues of Kharchia-65, thereby contributing to its improved salt-tolerance ability. Although TaHKT1;4 was expressed only in roots and was downregulated under the stress in salt-tolerant genotypes, it was not regulated through variations in cytosine methylation. Thus, understanding epigenetic regulation and the function of HKTs would enable an improvement in salt tolerance and the development of salt-tolerant crops.

Elicitor-Induced Biochemical and Molecular Manifestations to Improve Drought Tolerance in Rice (Oryza sativa L.) through Seed-Priming.

Rice (Oryza sativa L.) is one of the major grain cereals of the Indian subcontinent which face water-deficit stress for their cultivation. Seed-priming has been reported to be a useful approach to complement stress responses in plants. In the present study, seed-priming with hormonal or chemical elicitor [viz. methyl jasmonate (MJ), salicylic acid (SA), paclobutrazol (PB)] showed significant increase in total phenolic content, antioxidant activity and expression of Rice Drought-responsive (RD1 and RD2) genes (of AP2/ERF family) in contrasting rice genotypes (Nagina-22, drought-tolerant and Pusa Sugandh-5, drought-sensitive) under drought stress. However, decrease in lipid peroxidation and protein oxidation was observed not only under the stress but also under control condition in the plants raised from primed seeds. Expression analyses of RD1 and RD2 genes showed upregulated expression in the plants raised from primed seeds under drought stress. Moreover, the RD2 gene and the drought-sensitive genotype showed better response than that of the RD1 gene and the drought-tolerant genotype in combating the effects of drought stress. Among the elicitors, MJ was found to be the most effective for seed-priming, followed by PB and SA. Growth and development of the plants raised from primed seeds were found to be better under control and drought stress conditions compared to that of the plants raised from unprimed seeds under the stress. The present study suggests that seed-priming could be one of the useful approaches to be explored toward the development of simple, cost-effective and farmer-friendly technology to enhance rice yield in rainfed areas.