Antisense RNA approach targeting Rep gene of Mungbean yellow mosaic India virus to develop resistance in soybean

Abstract

The Yellow mosaic disease is caused by Mungbean yellow mosaic India virus (MYMIV) and Mungbean yellow mosaic virus (MYMV) belonging to the genus Begomovirus of the family Geminiviridae. Yellow mosaic disease (YMD) is a major constraint to the production of soybean in South-East Asia. In India, yield losses of 10–88% had been reported due to YMD of soybean. An effort has been made to generate resistant soybean plants, by a construct targeting replication initiation protein (Rep) gene sequences of MYMIV. A construct containing the sequences of Rep gene (566 bp) in antisense orientation was used to transform cotyledonary node explants of three soybean cultivars (JS 335, JS 95-60 and NRC 37). Transformation efficiencies of 0.2, 0.21 and 0.24% were obtained with three soybean cultivars, JS 335, JS 95-60 and NRC 37, respectively. The presence of transgene in T₁ plants was confirmed by polymerase chain reaction (PCR) and sequence analysis. The level of resistance was observed by challenge inoculation with the virus in T₁ lines. The inheritance of transgene showed classical Mendelian pattern in six transgenic lines.

Keywords:

• Begomovirus
• RNAi
• PDR
• yellow mosaic virus
• legumes

Acknowledgements

The financial support given by the Indian Council of Agricultural Research (ICAR), Government of India is gratefully acknowledged. We are thankful to Dr H.S. Gupta, Director, Indian Agricultural Research Institute, New Delhi-110012, India, Dr R.K. Jain, Head, Division of Plant Pathology, for providing the necessary facilities. We are also grateful to the scientists and staff at National Phytotron Facility, Indian Agricultural Research Institute, New Delhi-110012, India, for their guidance and support to grow plants under controlled conditions.