Shailender Kumar Verma

Transfer of useful variability of high grain iron and zinc from Aegilops kotschyi into wheat through seed irradiation approach

Abstract Purpose To transfer the 2S chromosomal fragment(s) of Aegilops kotschyi (2Sk) into the bread wheat genome which could lead to the biofortification of wheat with high grain iron and zinc content. Materials and methods Wheat-Ae. kotschyi 2A/2Sk substitution lines with high grain iron and zinc content were used to transfer the gene/loci for high grain Fe and Zn content into wheat using seed irradiation approach. Results Bread wheat plants derived from 40 krad-irradiated seeds showed the presence of univalents and multivalents during meiotic metaphase-I. Genomic in situ hybridization analysis of seed irradiation hybrid F2 seedlings showed several terminal and interstitial signals indicated the introgression of Ae. kotschyi chromosome segments. This proves the efficacy of seed radiation hybrid approach in gene transfer experiments. All the radiation-treated hybrid plants with high grain Fe and Zn content were analyzed with wheat group 2 chromosome-specific polymorphic simple sequence repeat markers to identify the introgression of small alien chromosome fragment(s). Conclusion Radiation-induced hybrids showed more than 65% increase in grain iron and 54% increase in Zn contents with better harvest index than the elite wheat cultivar WL711 indicating effective and compensating translocations of 2Sk fragments into wheat genome.

Uptake, distribution, and remobilization of iron and zinc among various tissues of wheat–Aegilops substitution lines at different growth stages

Biofortification of wheat for higher grain iron and zinc is the most feasible and cost-effective approach for alleviating micronutrient deficiency. The non-progenitor donor Aegilops species had 2–3 times higher grain iron and zinc content than the wheat cultivars, whereas the wheat–Aegilops substitution lines mostly of group 2 and 7 chromosomes had intermediate levels of grain micronutrients. The non-progenitor Aegilops species also had the highest iron content and intermediate-to-highest zinc content in straw, lower leaves, and flag leaves at the pre-anthesis, grain-filling, and maturity growth stages. The micronutrients accumulation status is followed by wheat–Aegilops substitution lines and is the least in wheat cultivars indicating that the donor Aegilops species and their substituted chromosomes possess genes for higher iron and zinc uptake and mobilization. The grain iron content was highly positively correlated with iron content in the plant tissues. Most of the lines had much higher iron and zinc content in all tissues during grain-filling period indicating higher iron and zinc uptake from soil during this stage. Although iron and zinc contents are nearly similar in grains, there was much less zinc content in the plant tissues of all the lines suggesting that the Triticeae species take up less zinc which is mobilized to grains more effectively than iron.

Proteome scale identification, classification and structural analysis of iron-binding proteins in bread wheat

Bread wheat is one of the major staple foods of worldwide population and iron plays a significant role in growth and development of the plant. In this report, we are presenting the genome wide identification of iron-binding proteins in bread wheat. The wheat genome derived putative proteome was screened for identification of iron-binding sequence motifs. Out of 602 putative iron-binding proteins, 130 were able to produce reliable structural models by homology techniques and further analyzed for the presence of iron-binding structural motifs. The computationally identified proteins appear to bind to ferrous and ferric ions and showed diverse coordination geometries. Glu, His, Asp and Cys amino acid residues were found to be mostly involved in iron binding. We have classified these proteins on the basis of their localization in the different cellular compartments. The identified proteins were further classified into their protein folds, families and functional classes ranging from structure maintenance of cellular components, regulation of gene expression, post translational modification, membrane proteins, enzymes, signaling and storage proteins. This comprehensive report regarding structural iron binding proteome provides useful insights into the diversity of iron binding proteins of wheat plants and further utilized to study their roles in plant growth, development and physiology.

Transferability and Polymorphism Between Group 7 Chromosome Specific Simple Sequence Repeat (SSR) Markers of Bread Wheat and Its Related Non-Progenitor Aegilops Species

ABSTRACT Simple sequence repeat (SSR) markers are highly efficient for genetic mapping and molecular breeding in crop plants. Chromosomes 7S and 7U of related non-progenitor Aegilops species possess superior genetic systems for high grain iron and zinc content. This study was undertaken to conduct a comprehensive analysis of transferability and polymorphism among group-7 anchored SSR markers of bread wheat (Triticum aestivum L.) to Aegilops species for wheat improvement. The study revealed 77% transferability of group 7-specific SSR markers of bread wheat to 7U/7S chromosomes of Aegilops species. More than 80% of the 7D specific markers were found to be transferable with a high level of polymorphism. The transferability to 7S and 7U genome suggested higher similarity between bread wheat genome(s) and S genome as compared with U genome of Aegilops species. These polymorphic markers are highly informative, robust and cost-effective for molecular breeding and could be further utilized for identification of introgression/transfer(s) of 7S/7U chromosomal fragments in bread wheat genome through wide hybridization.

Targeting metabolic pathways proteins of Orientia tsutsugamushi using combined hierarchical approach to combat scrub typhus.

Orientia tsutsugamushi (Ott) is a causative agent of chigger‐borne zoonosis, scrub typhus which is life threatening and highly pervasive illness in humans. In this report, we have mined and classified the proteins involved in pathways unique to Ott by using high‐throughput computational techniques. The 12 metabolic pathways were found to be unique to the pathogen. Forty‐six proteins were reported to be essential for the pathogens survival and non‐homologous to the humans. The proteins were categorized into different classes, ie, enzymes, transporters, DNA‐binding, secretory, and outer membrane proteins. Further, in silico analysis of 46 proteins showed that 25 proteins were suitable therapeutic targets with known druggable properties. The structural modeling of B3CSG3 (MurA) protein was carried out and catalytic site essential for its functioning was analyzed. Virtual screening of chemical compounds was performed against modeled structure. The docking study by AutodockVina reported compound from PubChem with CID: 16036947 as best and potential inhibitor by means of docking score and binding affinity. The reliability and stability of the MurA‐16036947 complex were confirmed with molecular dynamics simulation. The report will provide insight to understand the mechanism of pathogenesis of Ott and instigate the development of effective treatment strategies against this disease.

In silico Study of Iron, Zinc and Copper Binding Proteins of Pseudomonas syringae pv. lapsa: Emphasis on Secreted Metalloproteins

The phytopathogenic bacteria, Pseudomonas syringae pv. lapsa (P. syringae pv. lapsa) infects the staple food crop wheat. Metalloproteins play important roles in plant-pathogen interactions. Hence, the present work is aimed to predict and analyze the iron (Fe), zinc (Zn), and copper (Cu) binding proteins of P. syringae pv. lapsa which help in its growth, adaptation, survival and pathogenicity. A total of 232 Fe, 307 Zn, and 38 Cu-binding proteins have been identified. The functional annotation, subcellular localization and gene ontology enriched network analysis revealed their role in wide range of biological activities of the phytopathogen. Among the identified metalloproteins, a total of 29 Fe-binding, 31 Zn-binding, and 5 Cu-binding proteins were found to be secreted in nature. These putative secreted metalloproteins may perform diverse cellular and biological functions ranging from transport, response to oxidative stress, proteolysis, antimicrobial resistance, metabolic processes, protein folding and DNA repair. The observations obtained here may provide initial information required to draft new schemes to control microbial infections of staple food crops and will further help in developing sustainable agriculture.