Abstract
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.
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Abbreviations
- ANOVA:
-
Analysis of variance
- HD-ZIP III:
-
Homeodomain leucine zipper III
- MDA:
-
Malondialdehyde
- PEG:
-
Polyethylene glycol
- PGPR:
-
Plant growth promoting rhizobacteria
- RLM-RACE:
-
RNA ligase-mediated rapid 5′ amplification of cDNA ends
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Acknowledgements
The study was supported by the “Genome-wide editing for enhanced yield and quality traits” of CSIR Niche Creating High Science Project (MLP 0026) from the Council of Scientific and Industrial Research (C.S.I.R.), New Delhi, India. AY acknowledges a fellowship from the Department of Science and Technology, Government of India as a senior research fellowship [IF180146]. The authors also thank Dr. Puneet S. Chauhan for providing the PGPR, Pseudomonas putida bacterial strain (RA). CSIR-NBRI Manuscript No: CSIR-NBRI_MS/2022/05/03.
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Council for Scientific and Industrial Research, India, MLP0026, Indraneel Sanyal
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AY: conceptualized and performed the experiments. SK: was involved in writing, review & formal analysis. RV and SN: formally analysed and investigated the data. RJ, CL, SPR and PAS: critically reviewed the manuscript and provided guidance. The whole work was designed and performed under the supervision of Indraneel Sanyal from funding acquisition, review, editing and guidance for improving the manuscript. All authors have read and approved the manuscript.
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11240_2023_2458_MOESM1_ESM.pptx
Supplementary file1 Sequence similarity of miR166 and ATHB15 target sequence in chickpea and model plant, Arabidopsis thaliana (PPTX 50 KB)
11240_2023_2458_MOESM2_ESM.pptx
Supplementary file2 2.5 kb fragment of pmiR166 with BamHI and SacI used for double digestion to derive the 187 bp Car-miR166 gene fragment. 2B: PCR validation of transgenic lines (T0): M: marker (100 bp), 1-11: transgenic lines, C: negative control, NTC: no template control, P: positive control. CaMV35S F and miR166 R primer combinations were used for transgenic validation. (PPTX 423 KB)
11240_2023_2458_MOESM3_ESM.pptx
Supplementary file3 Different phytohormone applications (IAA, GA and ABA) on control plants andtransgenic lines (T1, T2 and T3) in presence of RA (PPTX 1402 KB)
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Yadav, A., Kumar, S., Verma, R. et al. Overexpression of PGPR responsive chickpea miRNA166 targeting ATHB15 for drought stress mitigation. Plant Cell Tiss Organ Cult 154, 381–398 (2023). https://doi.org/10.1007/s11240-023-02458-x
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DOI: https://doi.org/10.1007/s11240-023-02458-x