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Modulation of NPV gene expression pattern and retention of RNAi- based antiviral activity in inbred transgenic silkworm

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Abstract

Infections due to Bombyx mori nucleopolyhedro virus (BmNPV) in commercially important silkworm, Bombyx mori L. (Lepidoptera: Bombycidae) causes up to 50% loss to silk production in India. Increased resistance to NPV through introduction of double stranded RNA transgenes against multiple NPV genes is reported in B. mori. However, stability of the RNAi - mediated antiviral activity has not been determined after several generations of inbred transgenic B. mori. Hence, we examined the NPV multiplication rate and induction of the disease symptoms in NPV- infected transgenic and non-transgenic silkworm after 35 generations. Significantly higher multiplication rate of NPV supported by higher copy number of gp-41 gene was observed in non- transgenic larvae as compared to transgenic larvae. Expression of NPV genes, ie-1, lef1 and p74 enhanced significantly in non-transgenic larva as compared to transgenic larva after NPV infection. Strong positive correlation was observed in the expression pattern of NPV genes in transgenic larvae which was absent in non-transgenic larvae. This indicated co-regulation of NPV gene expression in the presence of transgenes.

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Acknowledgements

The authors express sincere thanks to the anonymous reviewers for critical reading and Central Silk Board, Bangalore for facilities and approval.

Funding

This work was supported by Department of Biotechnology (DBT), Government of India, New Delhi in the form of a research project on transgenic silkworm under Center of Excellence program (BT/01/COE/15/12-II dated 28/09/2011) to Center for DNA Fingerprinting & Diagnostics, Hyderabad and partially to Seribiotech Research Laboratory, Bangalore (AKA). VB is supported by research fellowship from the project.

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Authors and Affiliations

  1. Proteomics Division, Seribiotech Research Laboratory, CSB- Kodathi Campus, Carmelaram Post, Bangalore, 560035, Karnataka, India

    Burdekar Varada, Appukuttan Nair R. Pradeep, Arvind K. Awasthi & Kangayam M. Ponnuvel

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  1. Burdekar Varada
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  2. Appukuttan Nair R. Pradeep
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  3. Arvind K. Awasthi
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  4. Kangayam M. Ponnuvel
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Correspondence to Appukuttan Nair R. Pradeep.

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Supplementary Fig. 1

Genomic DNA of BmNPV purified from the NPV-infected B. mori larva was amplified using primers of the NPV genes ie-1, lef-1, lef-3, p-74 and gp-41 confirming NPV infection in early stage. Product size of the amplicons (in base pairs) is given in brackets. M- DNA ladder- Mass Ruler, Thermo). (PNG 3675 kb)

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Varada, B., Pradeep, A.N.R., Awasthi, A.K. et al. Modulation of NPV gene expression pattern and retention of RNAi- based antiviral activity in inbred transgenic silkworm. Int J Trop Insect Sci 40, 483–491 (2020). https://doi.org/10.1007/s42690-020-00201-z

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