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Transgenic expression of coat protein gene of Rice tungro bacilliform virus in rice reduces the accumulation of viral DNA in inoculated plants

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Abstract

Rice tungro, a devastating disease of rice in south and southeast Asia, is caused by the joint infection of Rice tungro bacilliform virus (RTBV) and Rice tungro spherical virus (RTSV). In order to obtain transgenic resistance against RTBV, indica rice cultivar Pusa Basmati-1 was transformed to express the coat protein (CP) gene of an Indian isolate of RTBV. Rice plants containing the transgene integrated in low copy numbers were obtained, in which the CP was shown to accumulate in the leaf tissue. The progenies representing three independent transformation events were challenged with Indian isolates of RTBV using viruliferous Green leafhoppers, and the viral titers in the inoculated plants were monitored using DNA dot-blot hybridization. As compared to non-transgenic controls, two independent transgenic lines showed significantly low levels of RTBV DNA, especially towards later stages of infection and a concomitant reduction of tungro symptoms.

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Acknowledgments

Authors acknowledge the expert technical assistance of Anand Singh Rana and efficient maintenance of plants and insects by Trilok Singh Rawat. This work was supported by grants from Department of Biotechnology, Government of India, New Delhi, to I.D. and M.V.R. (Grant no. BT/AB/NP/07/98). U.G. is grateful to Council of Scientific and Industrial Research, New Delhi, for the award of senior research fellowship.

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Authors and Affiliations

  1. Plant Polyamine and Transgenic Research Laboratory, Department of Genetics, University of Delhi, South Campus, New Delhi, 110021, India

    Uma Ganesan, Sarabjeet Singh Suri & Manchikatla Venkat Rajam

  2. Department of Plant Molecular Biology, University of Delhi, South Campus, New Delhi, 110021, India

    Shanmugam Rajasubramaniam & Indranil Dasgupta

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  1. Uma Ganesan
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  2. Sarabjeet Singh Suri
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  4. Manchikatla Venkat Rajam
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  5. Indranil Dasgupta
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Correspondence to Indranil Dasgupta.

Electronic supplementary material

11262_2009_359_MOESM1_ESM.pdf

Supplementary material 1 Absolute values of RTBV DNA levels expressed as densitometric readings of dot-blot hybridization using total DNA isolated from inoculated untransformed control and CP-60 transgenic plants taken at different days post-inoculation (PDF 22 kb)

11262_2009_359_MOESM2_ESM.pdf

Supplementary material 2 DNA from inoculated untransformed and transgenic rice plants used for dot-blot stained with methylene blue (left panel) and the same probed with RTBV DNA representing RT-RNase H region (right panel). C indicates DNA from untransformed control plants and T, transgenic line CP60. The numbers at the left indicate the days post-inoculation (dpi) at which the DNAs were extracted (PDF 15 kb)

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Ganesan, U., Suri, S.S., Rajasubramaniam, S. et al. Transgenic expression of coat protein gene of Rice tungro bacilliform virus in rice reduces the accumulation of viral DNA in inoculated plants. Virus Genes 39, 113–119 (2009). https://doi.org/10.1007/s11262-009-0359-9

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