Abstract
Key message
XPB2 and SEN1 helicases were identified through activation tagging as potential candidate genes in rice for inducing high water-use efficiency (WUE) and maintaining sustainable yield under drought stress.
Abstract
As a follow-up on the high-water-use-efficiency screening and physiological analyses of the activation-tagged gain-of-function mutant lines that were developed in an indica rice variety, BPT-5204 (Moin et al. in Plant Cell Environ 39:2440–2459, 2016a, https://doi.org/10.1111/pce.12796), we have identified two gain-of-function mutant lines (XM3 and SM4), which evidenced the activation of two helicases, ATP-dependent DNA helicase (XPB2) and RNA helicase (SEN1), respectively. We performed the transcript profiling of XPB2 and SEN1 upon exposure to various stress conditions and found their significant upregulation, particularly in ABA and PEG treatments. Extensive morpho-physiological and biochemical analyses based on 24 metrics were performed under dehydration stress (PEG) and phytohormone (ABA) treatments for the wild-type and the two mutant lines. Principal component analysis (PCA) performed on the dataset captured 72.73% of the cumulative variance using the parameters influencing the first two principal components. The tagged mutants exhibited reduced leaf wilting, improved revival efficiency, constant amylose:amylopectin ratio, high chlorophyll and proline contents, profuse tillering, high quantum efficiency and yield-related traits with respect to their controls. These observations were further validated under greenhouse conditions by the periodic withdrawal of water at the pot level. Germination of the seeds of these mutant lines indicated their insensitivity to high ABA concentration. The associated upregulation of stress-specific genes further suggests that their drought tolerance might be because of the coordinated expression of several stress-responsive genes in these two mutants. Altogether, our results provided a firm basis for SEN1 and XPB2 as potential candidates for manipulation of drought tolerance and improving rice performance and yield under limited water conditions.
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Abbreviations
- XPB2:
-
Xeroderma pigmentosa group B2
- SEN1:
-
T-RNA splicing endonuclease
- WUE:
-
Water-use efficiency
- PEG:
-
Polyethylene glycol
- ABA:
-
Abscisic acid
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Acknowledgments
MD is grateful to UoH BBL and UGC for the research fellowship and to DBT for funding the rice activation tagging project. MM is thankful to DST for the fellowship and research grant in the form of the INSPIRE-faculty program. AS and AB are grateful to DBT, and DD to DST-INSPIRE for proving Research Fellowships. MD is grateful to Dr. M.S. Madhav, Department of Biotechnology, Indian Institute of Rice Research, Hyderabad, India, for providing the infected rice samples and wild-type BPT-5204 seeds. PBK is grateful to the National Academy of Sciences-India for the NASI-Platinum Jubilee Senior Scientist award. The authors are grateful to Prof. M. Udaya Kumar, Bengaluru, India, for his help in IRMS analysis.
Funding
The activation-tagged rice population was developed in a DBT funded Project (BT/PR13105/Agr/02/684/2009) sanctioned to PBK while he was working at the University of Hyderabad. A part of the funding for this work is also obtained from the DST-INSPIRE faculty project (IFA17-LSPA67) sponsored to MM.
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PBK, MM and MD designed the experiments and prepared the manuscript. MD performed the experiments. MM and AB generated and maintained the activation-tagged mutant lines. MM performed the flanking sequence analysis through TAIL-PCR and identified the SEN1 and XPB2 genes. AS helped in the qRT-PCR experiments, analysis and seed counting. DD helped in PCA of the dataset. PBK supervised the work. MD, MM and PBK prepared the manuscript.
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Dutta, M., Moin, M., Saha, A. et al. Gain-of-function mutagenesis through activation tagging identifies XPB2 and SEN1 helicase genes as potential targets for drought stress tolerance in rice. Theor Appl Genet 134, 2253–2272 (2021). https://doi.org/10.1007/s00122-021-03823-0
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DOI: https://doi.org/10.1007/s00122-021-03823-0