Browsing by Author "Ankita Yadav"

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An overview on miRNA-encoded peptides in plant biology research
(Academic Press Inc., 2021) Ankita Yadav; Indraneel Sanyal; Shashi Pandey Rai; Charu Lata
MicroRNAs (miRNAs) are short (21–23 nt) regulatory RNA molecules present in plants and animals which are known for regulating the mRNA target gene expression either by cleavage or translational repression. With the advancements in miRNAs research in plants towards their biogenesis and applications has directed the recent discovery of pri-miRNAs encoding functional peptides or microRNA peptides (miPEPs). These miPEPs are encoded by 5′ of pri-miRs containing short ORFs (miORFs). miPEPs are known to enhance the activity of their associated miRNAs by increasing their accumulation and hence downregulating the target genes. Since miPEPs are very specific for each miRNA, they are considered as novel and effective tools for improving traits of interest for plant growth promotion and plant-microbe interaction. Entire peptidome research is the need of the hour. This review thus summarizes recent advancements in miPEP research and its applications as a technology with important agronomical implications with miRNAs augmentation. © 2021 Elsevier Inc.
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.
Development of a high-frequency in vitro regeneration system in Indian lotus (Nelumbo nucifera Gaertn.)
(Springer, 2024) Rita Verma; Ankita Yadav; Rajan Kumar Gupta; Indraneel Sanyal
The Indian lotus (Nelumbo nucifera Gaertn.) is a popular ornamental plant and a source of traditional herbal medicine. Its various parts are widely used in the pharmaceutical, cosmeceutical, and nutraceutical industries. The recent study aimed to develop a high-frequency in vitro regeneration system in Indian lotus. This study utilizes the pink lotus cultivar from the Botanical Garden of CSIR-NBRI Lucknow. The study was successfully achieved through direct and indirect methods using different plant growth regulators (PGRs). A direct regeneration system was established using explants shoot apical meristem and plumule cultured on SIM supplemented with 17 combinations and concentrations of BAP and NAA. Both explants produced the highest number of shoots with a combination of 4.44 μM BAP and 0.55 μM NAA. The highest number of shoots per explant 25 ± 1.0 was developed from the shoot apical meristem, while the plumule explant developed 16.3 ± 0.5 shoots per explant. Thereafter, the plantlets were transferred to LRIM, which contained 17 combinations and concentrations of NAA or IBA and BAP. The maximum number of roots, per explant 23.6 ± 0.5, was developed from shoot apical meristem using 2.22 μM NAA and 0.54 μM BAP. The highest number of roots, per explant 23 ± 1.0, was developed from the plumule using 4.44 μM IBA. Indirect somatic embryogenesis has been established through callus culture. The leaf segments were cultured onto a callus induction medium supplemented with ten combinations of 2,4-D and BAP. The high-frequency callus formation 24.33 ± 0.5 was obtained with a 5.0 μM 2,4-D and 1.0 μM BAP combination. All developmental stages at the proembryo, globular, heart, torpedo, and mature embryos were formed on concentrations of 2,4-D and BAP. After inducing shoot and root growth, well-developed plantlets were transferred to the greenhouse, resulting in a success rate of 18.47%. © The Society for In Vitro Biology 2024.
Effect of Stress-WIN, a novel polyherbal formulation, on DOCA-salt-induced hypertensive Wistar rats
(Springer Nature, 2025) Amit Ranjan; Anirban Roy; Poonam Pal; Somesh Agarwal; Amaresh Kumar Singh; Vinod N. Tiwari; Shreyans Kumar Jain; Hitesh Harsukhbhai Chandpa; Ankita Yadav; Sanjeev Kumar
Hypertension is a multifactorial disorder and one of the most important risk factors and a leading cause of stroke, heart disease, and end-organ damage. Hypertension leads to the production of high levels of pro-inflammatory cytokines and cholesterol, which increase thrombogenesis and fibrosis, ultimately resulting in chronic inflammation. Chronic inflammation causes endothelial dysfunction by producing excess reactive oxygen species (ROS) through pro-inflammatory cytokines such as interleukine-1β (IL-1β) and tumor necrosis factor-α (TNF-α). Further, the high level of nitric oxide (NO) and malonaldehyde (MDA) increases the oxidative stress, which worsens the liver's production of SGOT and SGPT and glomerular filtration of the kidney. In this study, an Ayurvedic polyherbal formulation (StressWIN) was shown to be a potent therapeutic drug for the treatment of deoxycorticosterone (DOCA)-salt-induced hypertension in Wistar rats. This study showed that administration of Stress-WIN significantly reduced the expression of pro-inflammatory cytokines (IL-1β and TNF-α) and also reduced the level of ROS, NO, MDA, SGOT, and SGPT in the serum of DOCA-salt-induced hypertension in Wistar rats. Furthermore, Stress-WIN treatment exhibited reduced cholesterol levels and glutathione concentration. Histological analysis showed that infiltration of immune cells was reversed by Stress-WIN A treatment (500 mg/kg and 1000 mg/kg). Furthermore, Stress-WIN A (2000 mg/kg) administration attenuated DOCA-salt-mediated morphological changes in the kidney and the heart of the Wistar rat. These promising outcomes underscore the potential of Stress-WIN as a viable alternative or adjunct therapy for hypertension, warranting further clinical investigations. © The Author(s) 2025.
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.
microRNA 166: an evolutionarily conserved stress biomarker in land plants targeting HD-ZIP family
(Springer, 2021) Ankita Yadav; Sanoj Kumar; Rita Verma; Charu Lata; Indraneel Sanyal; Shashi Pandey Rai
MicroRNAs (miRNAs) are significant class of noncoding RNAs having analytical investigating and modulatory roles in various signaling mechanisms in plants related to growth, development and environmental stress. Conserved miRNAs are an affirmation of land plants evolution and adaptation. They are a proof of indispensable roles of endogenous gene modulators that mediate plant survival on land. Out of such conserved miRNA families, is one core miRNA known as miR166 that is highly conserved among land plants. This particular miRNA is known to primarily target HD ZIP-III transcription factors. miR166 has roles in various developmental processes, as well as regulatory roles against biotic and abiotic stresses in major crop plants. Major developmental roles indirectly modulated by miR166 include shoot apical meristem and vascular differentiation, leaf and root development. In terms of abiotic stress, it has decisive regulatory roles under drought, salinity, and temperature along with biotic stress management. miR166 and its target genes are also known for their beneficial synergy with microorganisms in leguminous crops in relation to lateral roots and nodule development. Hence it is important to study the roles of miR166 in different crop plants to understand its defensive roles against environmental stresses and improve plant productivity by reprogramming several gene functions at molecular levels. This review is hence a summary of different regulatory roles of miR166 with its target HD-ZIP III and its modulatory and fine tuning against different environmental stresses in various plants. © 2021, Prof. H.S. Srivastava Foundation for Science and Society.
Overexpression of miR166 in Response to Root Rhizobacteria Enhances Drought Adaptive Efficacy by Targeting HD-ZIP III Family Genes in Chickpea
(Springer Science and Business Media Deutschland GmbH, 2024) Ankita Yadav; Sanoj Kumar; Rita Verma; Shiv Narayan; Uma Gupta; Charu Lata; Shashi Pandey Rai; Indraneel Sanyal
Using the transgenic approach, the current study investigated the tripartite interaction of miRNA166, Plant Growth Promoting Rhizobacteria (PGPR), and chickpea crops in response to drought. miR166, an evolutionarily conserved small RNA, was cloned and transformed in a homologous manner. This Car-miR166 is reported in our previous research to have drought-enduring roles in response to microbial candidates. A Pseudomonas putida strain RA (MTCC5279) is used as a PGPR for the whole study. The overexpressed lines generated using tissue-culture practice were functionally validated with physiological parameters studied using Li-Cor 6400XT, including photosynthesis rate, transpiration rate, water-use efficiency, and electron transport rate. We also studied the relative water content of the overexpressed lines in comparison to treated control plants. In biochemical methods, we studied the accumulation of proline, superoxide dismutase, peroxidase, catalase, H2O2 and lipid peroxidation levels. miR166 has its target as ATHB15 (Homeobox-leucine zipper protein-15) validated using 5’ RNA Ligase-Mediated Rapid Amplification of cDNA Ends (RLM-RACE) experiment. At the molecular levels, we carried out the stem-loop quantitative real-time (qRT) PCR analysis of miR166 and the expression analysis of ATHB15 in transgenic lines. As per our study, the results reported that the transgenic lines showed a positive interaction of miR166 with PGPR, resulting in drought stress mitigation and better plant survival in harsh drought conditions. In conclusion, the physiology, biochemistry, and molecular expression levels of Car-miR166 (Cicer arietinum L.) in transgenic lines in response to PGPR support enhanced growth and development in response to PGPR in transgenic lines under drought. © The Author(s) under exclusive licence to Sociedad Chilena de la Ciencia del Suelo 2024.
Overexpression of PGPR responsive chickpea miRNA166 targeting ATHB15 for drought stress mitigation
(Springer Science and Business Media B.V., 2023) Ankita Yadav; Sanoj Kumar; Rita Verma; Shiv Narayan; Ram Jatan; Charu Lata; Shashi Pandey Rai; Pramod A. Shirke; Indraneel Sanyal
Water limitation creates drought-like situations and constrains the life cycle of crop plants by modulating their biological processes at physiological, biochemical, and molecular levels. The microbial measures, including plant growth-promoting rhizobacteria (PGPR), could be used in plant adaptation. These PGPR escape water scarcity conditions and relates to plants by modulating several microRNAs in plant stress responses. The present study relates the beneficial role of PGPR (Pseudomonas putida-RA) responsive Car-miR166 of chickpea in drought mitigation with phytohormonal crosstalk in transgenic Arabidopsis lines. The transgenic lines showed an increased percentage of seed germination in comparison to treated control plants with highest germination rate in T2 (90%) and highest root length was observed in drought treated inoculated T1 lines (29%) under 300 mM of mannitol. The various physiological parameters including photosynthesis rate, transpiration rate, water-use efficiency and stomatal conductance were also better along with lower electrolyte leakage and higher relative water content in treated transgenic lines under inoculated conditions. The biochemical parameters including enzymatic and non-enzymatic antioxidants were improved in transgenic lines with less membrane damage and the highest accumulation of proline in T2 lines under RA inoculation and drought stress in comparison to treated control. The miR166 in drought-treated inoculated plants was highly upregulated (≥ 4) log2 fold change in T3 whereas the target was highly downregulated (≥ -2) log2 fold change in T2. Overall, our results concluded that RA-responsive Car-miR166 plays beneficial stress-mitigating roles under drought in transgenic plants, suggesting its crucial role in crop enhancement in response to PGPR. © 2023, The Author(s), under exclusive licence to Springer Nature B.V.
Target cleavage mapping and tissue-specific expression analysis of PGPR responsive miR166 under abiotic stress in chickpea (Cicer arietinum L.)
(Springer Science and Business Media B.V., 2023) Ankita Yadav; Sanoj Kumar; Rita Verma; Shashi Pandey Rai; Charu Lata; Indraneel Sanyal
Legumes are an indispensable food after cereals with extensive production across the world. Legume production is imposed with limitations and has been augmented by various environmental stresses. The symbiotic relations between legumes and rhizobacteria have been an intriguing topic of research in view of their roles in plant growth, development and various stress responses. Recent advances in gene networks involving a plethora of evolutionarily conserved miRNAs have been investigated pertaining to their roles in plant stress responses. The interaction between plant growth-promoting rhizobacteria (PGPR) strain Pseudomonas putida (RA), MTCC5279 and abiotic stress-responsive miRNAs have previously been studied with roles in abiotic stress mitigation by modulating stress-responsive miRNAs and their target genes. The present study is an investigation involving the role of RA-responsive miR166 for drought mitigation in desi chickpea genotype. Drought-stressed chickpea plants when inoculated with RA, the inverse correlation in expression patterns were noticed in miR166 and its validated target, ATHB15. miR166-directed cleavage of ATHB15 has been carried out in drought-treated plantlets upon RA inoculation using 5´RLM-RACE analysis. Tissue-specific expression patterns in 15 days old chickpea seedlings including leaves, shoot and roots when exposed to salinity, drought and abscisic acid at different time points indicating the role of miR166 in different abiotic stress responses. In view of the results, validation and functional characterization of such interactions involving stress-responsive miRNAs along with microbial applications in stress management could be an important method for crop improvement. © 2023, The Author(s), under exclusive licence to Springer Nature B.V.