Abstract
Winter survival develops efficient tolerance mechanisms in plants by regulating cold-responsive and cold-regulated genes at the transcriptional level. Hence, an insight into the expression would provide the molecular function of these cold responsive genes. In this study, an uncharacterized gene encoding a cold-regulated (Cor413) protein identified from Saccharum spontaneum (wild relative species of sugarcane) low-temperature transcriptome with environmental adaptability is isolated and characterized. The full-length coding region possesses an open reading frame of 642 bp, which encodes a putative polypeptide of 213 amino acids of molecular weight 25.6 kDa and an isoelectric point (Pi) of 9.69. The SsCor413 sequence showed a high similarity to monocot Cor413 proteins comprising a WCOR413 domain. Bioinformatics analysis revealed that Cor413 protein has multispanning transmembrane helices along with highly conserved phosphorylation sites. String analysis suggested that SsCor413 is grouped with LEA and Rab proteins that are involved in freezing tolerance. Gene ontology analysis assigned the protein to terms such as “plasma membrane,” “cold acclimation,” and “response to cold.” Sub-cellular localization experiments of sugarcane callus and onion epidermal cells indicated the nuclear localized expression. Quantitative gene expression analysis indicated that the SsCor413 gene is up-regulated in leaf and root tissues of S. spontaneum under low temperature, salinity, and water deficit stress conditions. These results highlight the potential role of SsCor413 in abiotic stress tolerance, and this gene could be a new candidate for combating multiple stresses in sugarcane.
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Abbreviations
- Cor:
-
Cold regulated
ORF
Open reading frame
GFP
Green Fluorescent Protein
LEA
Late Embryogenic Abundant Protein
GO
Gene Ontology
LT
Low temperature
MS
Murashige and Skoog
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Acknowledgments
The authors would like to thank ICAR—Sugarcane Breeding Institute, Coimbatore for providing the necessary infrastructure. We thank Dr. Shobakumari, ICAR-SBI for extending microscope facility. We greatly acknowledge Dr. K. Kadirvelu, DRDO-BU, Coimbatore to access the fluorescence microscope. Thanks to Mr. K. Selvamuthu for his technical assistance to carry out the work.
Funding
The authors would like to thank Science and Engineering Research Board (SERB), Department of Science and Technology (DST), New Delhi (Grant No SB/YS/LS-165/2013) for financial support. Authors thank the Department of Science and Technology, India (DST-INSPIRE, IF150891) for financial support to DS.
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DS and AC designed the experiments. DS performed the experiments and wrote the manuscript. SGS provided localization control vectors, assisted in microscopic studies, and manuscript revision. MVM and ANJ supported in cloning. SPTS did the artwork for figs. RK and MRM helped in bioinformatics analysis. MM, BR, and AC revised the manuscript. All authors read and approved the final manuscript.
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Key Message
Cor413 is identified as abiotic stress responsive gene in Saccharum spontaneum and acts as potential gene for combating abiotic stresses in sugarcane.
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Dharshini, S., Manoj, V.M., Suresha, G.S. et al. Isolation and Characterization of Nuclear Localized Abiotic Stress Responsive Cold Regulated Gene 413 (SsCor413) from Saccharum spontaneum. Plant Mol Biol Rep 38, 628–640 (2020). https://doi.org/10.1007/s11105-020-01224-z
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DOI: https://doi.org/10.1007/s11105-020-01224-z