Impact of foliar application of iron and zinc fertilizers on grain iron, zinc, and protein contents in bread wheat (Triticum aestivum L.).

Introduction: Micronutrient deficiencies, particularly iron (Fe) and zinc (Zn), are prevalent in a large part of the human population across the world, especially in children below 5 years of age and pregnant women in developing countries. Since wheat constitutes a significant proportion of the human diet, improving grain Fe and Zn content in wheat has become important in improving human health. Objective: This study aimed to quantify the effect of foliar application of iron sulfate heptahydrate (FeSO4.7H2O) and zinc sulfate heptahydrate (ZnSO4.7H2O) and their combination on grain Fe and Zn concentrations, as well as grain protein content (GPC). The study also aimed to assess the utility of these applications in large field conditions. Methods: To address this issue, field experiments were conducted using 10 wheat cultivars and applying a foliar spray of FeSO4.7H2O (0.25%) and ZnSO4.7H2O (0.50%) separately (@400 L of solution in water per hectare during each spray) and in combination at two different crop growth stages (flowering and milking) for three consecutive crop seasons (2017-2020). The study used a split-plot design with two replications to assess the impact of foliar application on GFeC, GZnC, and GPC. In addition, an experiment was also conducted to assess the effect of soil (basal) @ 25 kg/ha ZnSO4, foliar @ 2 kg/ha, ZnSO4.7H2O (0.50%), and the combination of basal + foliar application of ZnSO4 on the grain micronutrient content of wheat cultivar WB 02 under large field conditions. Results: GFeC increased by 5.1, 6.1, and 5.9% with foliar applications of FeSO4, ZnSO4, and their combination, respectively. GZnC increased by 5.2, 39.6, and 43.8% with foliar applications of FeSO4, ZnSO4, and their combination, respectively. DBW 173 recorded the highest increase in GZnC at 56.9% with the combined foliar application of FeSO4 and ZnSO4, followed closely by HPBW 01 at 53.0% with the ZnSO4 foliar application, compared to the control. The GPC increased by 6.8, 4.9, and 3.3% with foliar applications of FeSO4, ZnSO4, and their combination, respectively. Large-plot experiments also exhibited a significant positive effect of ZnSO4 not only on grain Zn (40.3%, p ≤ 0.001) and protein content (p ≤ 0.05) but also on grain yield (p ≤ 0.05) and hectoliter weight (p ≤ 0.01), indicating the suitability of the technology in large field conditions. Conclusion: Cultivars exhibited a slight increase in GFeC with solitary foliar applications of FeSO4, ZnSO4, and their combination. In contrast, a significant increase in GZnC was observed with the foliar application of ZnSO4 and the combined application of FeSO4 and ZnSO4. In terms of GPC, the most significant enhancement occurred with the foliar application of FeSO4, followed by ZnSO4 and their combination. Data demonstrated the significant effect of foliar application of ZnSO4 on enhancing GZnC by 39.6%. Large plot experiments also exhibited an increase of 40.3% in GZnC through the foliar application of ZnSO4, indicating the effectiveness of the technology to be adopted in the farmer's field.

Utilization of Grain Physical and Biochemical Traits to Predict Malting Quality of Barley (Hordeum vulgare L.) under Sub-Tropical Climate.

Barley is the most popular raw material for malting, and recently, the demand for malt-based products has increased several folds in India and other South Asian countries. The barley growing season is peculiar in the sub-tropical plains region compared to European or Northern American conditions, characterized by a total crop duration of 130-145 days with a maximum grain filling duration of around only 35-40 days. A total of 19 barley genotypes were grown for three years to assess the comparative performance in relation to different quality traits, including grain physical traits and biochemical and malt quality parameters. Analysis of variance, Pearson correlation, and principal component analysis were performed to determine the correlation among different traits. The results showed significant genotypic variation among genotypes for individual grain and malt traits. Despite the shorter window for grain filling, several good malting genotypes have been developed for the sub-tropical climates. The genotypes DWRUB52, DWRB101, RD2849, DWRUB64, and DWRB91 were found suitable for malting. Based on correlation studies, a few grain parameters have been identified which can be used to predict the malting potential of a barley genotype. The hot water extract was found to be positively correlated with the grain test weight, thousand-grain weight, and malt friability but was negatively correlated with the husk content. Beta-glucan content varied from 3.4 to 6.1% (dwb); reducing the grain beta-glucan content and increasing the amylase could be priorities to address in future malt barley improvement programs under sub-tropical climatic conditions.

GWAS to Identify Novel QTNs for WSCs Accumulation in Wheat Peduncle Under Different Water Regimes.

Water-soluble carbohydrates (WSCs) play a vital role in water stress avoidance and buffering wheat grain yield. However, the genetic architecture of stem WSCs' accumulation is partially understood, and few candidate genes are known. This study utilizes the compressed mixed linear model-based genome wide association study (GWAS) and heuristic post GWAS analyses to identify causative quantitative trait nucleotides (QTNs) and candidate genes for stem WSCs' content at 15 days after anthesis under different water regimes (irrigated, rainfed, and drought). Glucose, fructose, sucrose, fructans, total non-structural carbohydrates (the sum of individual sugars), total WSCs (anthrone based) quantified in the peduncle of 301 bread wheat genotypes under multiple environments (E01-E08) pertaining different water regimes, and 14,571 SNPs from "35K Axiom Wheat Breeders" Array were used for analysis. As a result, 570 significant nucleotide trait associations were identified on all chromosomes except for 4D, of which 163 were considered stable. A total of 112 quantitative trait nucleotide regions (QNRs) were identified of which 47 were presumable novel. QNRs qWSC-3B.2 and qWSC-7A.2 were identified as the hotspots. Post GWAS integration of multiple data resources prioritized 208 putative candidate genes delimited into 64 QNRs, which can be critical in understanding the genetic architecture of stem WSCs accumulation in wheat under optimum and water-stressed environments. At least 19 stable QTNs were found associated with 24 prioritized candidate genes. Clusters of fructans metabolic genes reported in the QNRs qWSC-4A.2 and qWSC-7A.2. These genes can be utilized to bring an optimum combination of various fructans metabolic genes to improve the accumulation and remobilization of stem WSCs and water stress tolerance. These results will further strengthen wheat breeding programs targeting sustainable wheat production under limited water conditions.

Comparative physiological, biochemical and transcriptomic analysis of hexaploid wheat (T. aestivum L.) roots and shoots identifies potential pathways and their molecular regulatory network during Fe and Zn starvation.

The combined effect of iron (Fe) and zinc (Zn) starvation on their uptake and transportation and the molecular regulatory networks is poorly understood in wheat. To fill this gap, we performed a comprehensive physiological, biochemical and transcriptome analysis in two bread wheat genotypes, i.e. Narmada 195 and PBW 502, differing in inherent Fe and Zn content. Compared to PBW 502, Narmada 195 exhibited increased tolerance to Fe and Zn withdrawal by significantly modulating the critical physiological and biochemical parameters. We identified 25 core genes associated with four key pathways, i.e. methionine cycle, phytosiderophore biosynthesis, antioxidant and transport system, that exhibited significant up-regulation in both the genotypes with a maximum in Narmada 195. We also identified 26 microRNAs targeting 14 core genes across the four pathways. Together, core genes identified can serve as valuable resources for further functional research for genetic improvement of Fe and Zn content in wheat grain.

Transcriptomic dataset reveals the molecular basis of genotypic variation in hexaploid wheat (T. aestivum L.) in response to Fe/Zn deficiency.

The datasets depicted in the paper are related to the original article entitled "Identifying transcripts associated with efficient transport and accumulation of Fe and Zn in hexaploid wheat (T. aestivum L.)" [1]. Four wheat genotypes i.e. Sonora 64, CRP 1660, Vinata, and DBW 17 were selected for RNA sequencing using Illumina HiSeq4000 platform. These genotypes were grown in Fe/Zn sufficient and deficient conditions in sand pot culture with intermittent administration of Hoagland solution. Pooled assembly was carried out for all of the four varieties subsequent to discarding low-quality reads, adaptor sequences and contamination resulting in approximately 315,904 clean transcripts of around 937 bp lengths and N50 of 1,294 bp. For the functional annotation of the identified transcripts databases like Pfam, KEGG pathway, Uniprot, PlnTFDB and wheat proteins were utilized. Differential expression calculation of transcripts was carried out by DESeq, an R package and real-time PCR study of 12 Fe/Zn metabolic pathway related transcripts was utilized for further revalidation of data. Elemental analysis of grain Fe and Zn was performed using Flame Atomic Absorption Spectrometry (FAAS). The RNA-seq data of all the four wheat genotypes was uploaded on Sequence Read Archive (SRA: SUB6961770 and BioProject: PRJNA605909), enabling easy access to the researchers worldwide.

Identifying transcripts associated with efficient transport and accumulation of Fe and Zn in hexaploid wheat (T. aestivum L.).

Wheat (T. aestivum L.) is the second most important staple food crop consumed in the form of various end-use products across the world. However, it contains lower concentrations of Fe and Zn leading to micronutrient deficiency in human beings where wheat is the sole diet. Therefore, increasing grain Fe/Zn content in wheat has become priority in wheat breeding programmes across the world. Understanding the molecular mechanism of Fe/Zn transport and accumulation in grains is required to expedite the breeding process. For this purpose, whole seedling transcriptome analysis was conducted in four wheat genotypes (CRP 1660, Sonora 64, Vinata, : high, and DBW17: low) differing in grain Fe/Zn content under controlled and Fe/Zn deficient conditions. Twenty eight key transcripts involved in phytosiderophore biosynthesis, Fe/Zn uptake and transport were identified. Expression analysis of 12 of the transcripts using qPCR was conducted in seedling stage and flag leaf which exhibited greater differential accumulation in CRP 1660 followed by Vinata, Sonora 64 and DBW 17 in both flag leaf and seedling. However, there was significantly higher differential accumulation of the transcripts in flag leaf as compared to seedling. In CRP 1660, transcripts pertaining to phytosiderophore biosynthesis like DMAS1-B, NRAMP2 and NAAT2-D showed greater accumulation. Additionally, corresponding miRNAs were also identified for these 28 transcripts. The findings will help in better understanding of molecular basis of Fe/Zn transport and accumulation in grain and subsequent utilization in breeding to improve Fe/Zn content in wheat grain.

Hulless barley as a promising source to improve the nutritional quality of wheat products.

In this study, efforts were made to utilize hulless barley (variety BHS352) to enhance the nutritive value of chapatti and biscuit made from wheat flour. Barley flour was added to wheat flour in different ratios (5 to 30%). Antioxidant activity, total phenolic content and ß-glucan content were determined both in flour blends and their products. Changes in physical quality and taste of chapatti and biscuits after blending of hulless barley flour with wheat flour were measured. The chapatti quality score decreased by 15% and biscuit spread factor by 33% after 30% barley flour blending. Significant increase in ß-glucan content and antioxidant activity of flour blends and their products was observed at 30% blending level. The phenolic content increased from 63 to 135 µg for biscuits and 237 to 287 ug GAE/g for chapatti with blending of 30% barley flour.

Differential Activity and Expression Profile of Antioxidant Enzymes and Physiological Changes in Wheat (Triticum aestivum L.) Under Drought.

Wheat crop may experience water deficit at crucial stages during its life cycle, which induces oxidative stress in the plants. The antioxidant status of the plant plays an important role in providing tolerance against the water stress. The objective of this study was to investigate the impact of water stress on physiological traits, antioxidant activity and transcript profile of antioxidant enzyme related genes in four wheat genotypes (C306, AKAW3717, HD2687, PBW343) at three crucial stages of plants under medium (75% of field capacity) and severe stress (45% of field capacity) in pots. Drought was applied by withholding water for 10 days at a particular growth stage viz. tillering, anthesis and 15 days after anthesis (15DAA). For physiological traits, a highly significant effect of water stress at a particular stage and genotypic variations for resistance to drought tolerance was observed. Under severe water stress, the malondialdehyde (MDA) content increased while the relative water content (RWC) and chlorophyll index decreased significantly in all the genotypes. The drought susceptibility index (DSI) of the genotypes varied from 0.18 to 1.9. The drought treatment at the tillering and anthesis stages was found more sensitive in terms of reduction in thousand grain weight (TGW) and grain yield. Antioxidant enzyme activities [superoxide dismutase (SOD) and peroxidase (POX)] increased with the decrease in osmotic potential in drought tolerant genotypes C306 and AKAW3717. Moreover, the transcript profile of Mn-SOD upregulated significantly and was consistent with the trend of the variation in SOD activity, which suggests that Mn-SOD might play an important role in drought tolerance.