Browsing by Author "Ankesh Pandey"

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Correction to: Expression of the entomotoxic Cocculus hirsutus trypsin inhibitor (ChTI) gene in transgenic chickpea enhances its underlying resistance against the infestation of Helicoverpa armigera and Spodoptera litura (Plant Cell, Tissue and Organ Culture (PCTOC), (2021), 146, 1, (41-56), 10.1007/s11240-021-02041-2)
(Springer Science and Business Media B.V., 2021) Ankesh Pandey; Reena Yadav; Sanoj Kumar; Anil Kumar; Priya Shukla; Ankita Yadav; Indraneel Sanyal
The following sections were omitted in the initial online publication: Acknowledgements, Author contributions, and Declaration. The original article has been corrected. © 2021, Springer Nature B.V.
Expression of the entomotoxic Cocculus hirsutus trypsin inhibitor (ChTI) gene in transgenic chickpea enhances its underlying resistance against the infestation of Helicoverpa armigera and Spodoptera litura
(Springer Science and Business Media B.V., 2021) Ankesh Pandey; Reena Yadav; Sanoj Kumar; Anil Kumar; Priya Shukla; Ankita Yadav; Indraneel Sanyal
A trypsin inhibitor from Cocculus hirsutus, commonly known as “Farid Buti” has been demonstrated to exhibit insecticidal, fungicidal, as well as nematocidal activity. The ChTI (Cocculus hirsutus Trypsin Inhibitor) gene was designed in silico and synthesized by PCR-based gene synthesis and cloned in the plant expression vector pBI121, with kanamycin, as the selectable marker. Agrobacterium strain LBA4404 was transformed with pBI121:ChTI vector for plant transformation. For developing insect-tolerant chickpea, Agrobacterium-mediated transformation of ChTI gene was performed in cultivar P-362. Twenty-day-old cotyledonary node (CN) explants were used for sonication-assisted Agrobacterium-mediated transformation. Three cycles of increasing concentrations of kanamycin were used for the selection of transformed shoots. In vitro grown transgenic chickpea shoots were grafted on decapitated stock of chickpea seedlings. After 45–50 days of acclimatization and hardening, pod development and its maturation occurred. After screening by PCR, seven transgenic events were confirmed to be positive by Southern blot hybridization analysis, showing 1–4 copies of the transgene. The quantitative expression of the ChTI gene by qRT-PCR analysis showed up to 12–17-fold change in the T1 progeny. Immunoblot analysis revealed the expression of 31 kDa and 15 kDa ChTI protein in E.coli and transgenic plants respectively. Trypsin activity assay was performed in the T1 generation and higher anti-trypsin activity was recorded. Insect tolerance against Helicoverpa armigera and Spodoptera litura were estimated by insect bioassay, wherein an overall mortality of 60–80% and weight loss (30–60% and 40–60% for Spodoptera litura and Helicoverpa armigera respectively) have been recorded in the plants of T1 generation. © 2021, The Author(s), under exclusive licence to Springer Nature B.V. part of Springer Nature.
Genome-wide profiling of drought-tolerant Arabidopsis plants over-expressing chickpea MT1 gene reveals transcription factors implicated in stress modulation
(Springer Science and Business Media Deutschland GmbH, 2022) Sanoj Kumar; Ankita Yadav; Nasreen Bano; Arvind Kumar Dubey; Rita Verma; Ankesh Pandey; Anil Kumar; Sumit Bag; Sudhakar Srivastava; Indraneel Sanyal
Drought, a major abiotic limiting factor, could be modulated with in-built reprogramming of plants at molecular level by regulating the activity of plant developmental processes, stress endurance and adaptation. The transgenic Arabidopsis thaliana over-expressing metallothionein 1 (MT1) gene of desi chickpea (Cicer arietinum L.) was subjected to transcriptome analysis. We evaluated drought tolerance of 7 days old plants of Arabidopsis thaliana in both wild-type (WT) as well as transgenic plants and performed transcriptome analysis. Our analysis revealed 24,737 transcripts representing 24,594 genes out of which 5,816 were differentially expressed genes (DEGs) under drought conditions and 841 genes were common in both genotypes. A total of 1251 DEGs in WT and 2099 in MT1 were identified in comparison with control. Out of the significant DEGs, 432 and 944 were upregulated, whereas 819 and 1155 were downregulated in WT and MT1 plants, respectively. The physiological and molecular parameters involving germination assay, root length measurements under different stress treatments and quantitative expression analysis of transgenic plants in comparison to wild-type were found to be enhanced. CarMT1 plants also demonstrated modulation of various other stress-responsive genes that reprogrammed themselves for stress adaptation. Amongst various drought-responsive genes, 24 DEGs showed similar quantitative expression as obtained through RNA sequencing data. Hence, these modulatory genes could be used as a genetic tool for understanding and delineating the mechanisms for fine-tuning of stress responses in crop plants. © 2021, The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.
Metallothionein (MT1): A molecular stress marker in chickpea enhances drought and heavy metal stress adaptive efficacy in transgenic plants
(Elsevier B.V., 2022) Sanoj Kumar; Ankita Yadav; Rita Verma; Arvind Kumar Dubey; Shiv Narayan; Ankesh Pandey; Anshu Sahu; Sudhakar Srivastava; Indraneel Sanyal
Metallothioneins (MTs) are diverse class of cysteine-rich proteins having metal-chelation properties. The role of MTs has been demonstrated in different abiotic stresses and MTs have been designated as biomolecular stress markers. Chickpea is an important legume crop supplying proteins to humans, as well as acting as great soil-binders along with nitrogen-fixation capability. The present research deals with the development of transgenic chickpea overexpressing metallothionein type-1 (CarMT1) gene for analyzing its role in stress tolerance against drought and heavy metals. The overexpression construct was designed using binary expression vector, pBI121 and transformed in chickpea desi cultivar, Pusa-362 for functional validation by using sonication-assisted Agrobacterium-mediated transformation. The results indicated high transcript levels under the drought (22-folds) and changes in physiological (photosynthesis rate, transpiration rate, stomatal conductance, water-use efficiency) and biochemical (antioxidant enzymes and compatible solutes) parameters suggesting stress-mitigating roles of CarMT1. The transgenic seeds were evaluated for heavy metal stress adaptation that resulted in better seed survival efficiency under different heavy metal stresses. The results indicated beneficial roles of MT gene in transgenic lines of chickpea in presence of different abiotic stresses, which could pave the way for multi-stress tolerant crop development. © 2022 Elsevier B.V.