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StCaM2, a calcium binding protein, alleviates negative effects of salinity and drought stress in tobacco

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Overexpression of StCaM2 in tobacco promotes plant growth and confers increased salinity and drought tolerance by enhancing the photosynthetic efficiency, ROS scavenging, and recovery from membrane injury.

Abstract

Calmodulins (CaMs) are important Ca2+ sensors that interact with effector proteins and drive a network of signal transduction pathways involved in regulating the growth and developmental pattern of plants under stress. Herein, using in silico analysis, we identified 17 CaM isoforms (StCaM) in potato. Expression profiling revealed different temporal and spatial expression patterns of these genes, which were modulated under abiotic stress. Among the identified StCaM genes, StCaM2 was found to have the largest number of abiotic stress responsive promoter elements. In addition, StCaM2 was upregulated in response to some of the selected abiotic stress in potato tissues. Overexpression of StCaM2 in transgenic tobacco plants enhanced their tolerance to salinity and drought stress. Accumulation of reactive oxygen species was remarkably decreased in transgenic lines compared to that in wild type plants. Chlorophyll a fluorescence analysis suggested better performance of photosystem II in transgenic plants under stress compared to that in wild type plants. The increase in salinity stress tolerance in StCaM2-overexpressing plants was also associated with a favorable K+/Na+ ratio. The enhanced tolerance to abiotic stresses correlated with the increase in the activities of anti-oxidative enzymes in transgenic tobacco plants. Overall, our results suggest that StCaM2 can be a novel candidate for conferring salt and drought tolerance in plants.

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Acknowledgements

Dr. Deepak Kumar is thankful to the Science and Engineering Research Board (Grant No. EEQ/2016/000487), New Delhi, Government of India for providing the financial support.

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Authors and Affiliations

  1. Department of Botany, Central University of Jammu, Rahya-Suchani (Bagla), Dist- Samba, Jammu and Kashmir, 181143, India

    Meenakshi Raina & Deepak Kumar

  2. Plant Molecular Biology Laboratory, Faculty of Life Sciences and Biotechnology, South Asian University, Akbar Bhawan, Chanakyapuri, New Delhi, 110021, India

    Ashish Kumar, Nikita Yadav & Ananda Mustafiz

  3. Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, Jammu and Kashmir, India

    Sumita Kumari

  4. Department of Bioengineering, Integral University, Dasauli, Kursi Road, Lucknow, 226026, India

    Mohd Aslam Yusuf

  5. Department of Botany, Institute of Science, Banaras Hindu University, Varanasi, 221005, Uttar Pradesh, India

    Deepak Kumar

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Contributions

The work presented here was carried out in collaboration among all the authors. DK, SK, and AM conceived and designed the research. MR contributed to cloning and functional validation experiments in plants. AK contributed to Bioinformatics experiments and NY and MR conducted the real-time experiments and analysed the data. MR, AK, and DK wrote the manuscript, and MAY provided vital advice on the article and edited the manuscript. All authors read and approved the final manuscript.

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Correspondence to Ananda Mustafiz or Deepak Kumar.

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11103_2021_1131_MOESM1_ESM.pptx

Supplementary Fig. S1 Wordcloud based on results from PLACE database. (a) Using the keywords associated with promoter elements present in the StCaMgenes. (b) Using promoter motif names. Larger word size means larger representation in the promoters. Supplementary Fig. S2 RNA-seq based expression profiling of StCaMs. Expression heatmap of StCaM genes under (a) abiotic stress, (b) hormone treatment and (c) different tissues. FPKM values of StCaM genes were scaled and centered before plotting the heatmap. Clustering in the heatmap is based on correlation. Supplementary Fig. S3 Generation and molecular analysis of transgenic tobacco plants expressing the StCaM2 gene. (a) T-DNA portion of the pCAM-StCaM2 vector construct used for tobacco transformation. (b) PCR analysis to screen the transgenic plants for the presence of transgene was done using StCaM2 gene-specific primers. (c) Southern blot analysis showing the integration and copy number of the transgene. (d) RT-PCR analysis confirming expression of StCaM2 in young fully expanded leaves of transgenic tobacco plants. Actin gene was used as an internal control. Wild-type (WT) tobacco control. (C1, C3, C4, C6, C7, and C9) independently transformed T1 transgenic lines of tobacco (PPTX 1043 KB)

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Raina, M., Kumar, A., Yadav, N. et al. StCaM2, a calcium binding protein, alleviates negative effects of salinity and drought stress in tobacco. Plant Mol Biol 106, 85–108 (2021). https://doi.org/10.1007/s11103-021-01131-1

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