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Engineering resistance against Cucumber mosaic virus in Nicotiana tabacum through virus derived transgene expressing hairpin RNA

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Abstract

Cucumber mosaic virus (CMV) is the one of notorious virus known for its ubiquitous nature and causes substantial yield loss worldwide. The resistance against the Cucumber mosaic virus (CMV) was envisaged in Nicotiana tabacum transgenic lines by introducing viral gene fragments. The chimeric hairpin RNA constructs incorporating 401 bp of coat protein, 411 bp of replicase protein and 361 bp of 2b gene were developed respectively and transformed into N. tabacum. The regenerated transgenic lines introduced with inverted repeats of CMV gene fragments exhibited enhanced resistance against CMV. The preliminary molecular screening and qPCR confirmed the integration of transgene in the transgenic lines. The spectrum of resistance in transgenic lines was evaluated by challenge inoculation with CMV and the resistance was determined through DAC-ELISA. The complete resistance was achieved in the hpRNA-CP transformant with a very low titre (0.029) of CMV followed by hpRNA-REP (0.099) with no symptoms.

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Acknowledgements

We are thankful to Department of Plant Pathology, TNAU, Coimbatore for providing facilities during the course of study. We would like to extend our gratitude to CISRO Plant Industry, Australia and Dr. Varanavasiappan, Asst. Professor, Department of Biotechnology, TNAU for their immense help and support.

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Authors and Affiliations

  1. Department of Plant Pathology, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu, 641003, India

    J. Vinodhini, L. Rajendran & G. Karthikeyan

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  1. J. Vinodhini
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  2. L. Rajendran
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  3. G. Karthikeyan
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Contributions

GK designed the project, supervised, revised and approved the manuscript for submission; JV drafted the experiments and prepared the manuscript; LR gave proposition to conduct the experiment. All authors reviewed and approved the submission.

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Correspondence to G. Karthikeyan.

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Vinodhini, J., Rajendran, L. & Karthikeyan, G. Engineering resistance against Cucumber mosaic virus in Nicotiana tabacum through virus derived transgene expressing hairpin RNA. 3 Biotech 13, 143 (2023). https://doi.org/10.1007/s13205-023-03576-1

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