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Mungbean yellow mosaic virus (MYMV) AC4 suppresses post-transcriptional gene silencing and an AC4 hairpin RNA gene reduces MYMV DNA accumulation in transgenic tobacco

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Abstract

Mungbean yellow mosaic virus (MYMV) is a legume-infecting geminivirus that causes yellow mosaic disease in blackgram, mungbean, soybean, Frenchbean and mothbean. AC4/C4, which is nested completely within the Rep gene, is less conserved among geminiviruses. Much less is known about its role in viral pathogenesis other than its known role in the suppression of host-mediated gene silencing. Transient expression of MYMV AC4 by agroinfiltration suppressed post-transcriptional gene silencing in Nicotiana benthamiana 16c expressing green fluorescence protein, at a level comparable to MYMV TrAP expression. AC4 full-length gene and an inverted repeat of AC4 (comprising the full-length AC4 sequence in sense and antisense orientations with an intervening intron) which makes a hairpin RNA (hpRNA) upon transcription were introduced into tobacco by Agrobacterium-mediated leaf disc transformation. Leaf discs of the transgenic plants were agroinoculated with partial dimers of MYMV and used to study the effect of the AC4-sense and AC4 hpRNA genes on MYMV DNA accumulation. Leaf discs of two transgenic plants that express the AC4-sense gene displayed an increase in MYMV DNA accumulation. Leaf discs of six transgenic plants containing the AC4 hpRNA gene accumulated small-interfering RNAs (siRNAs) specific to AC4, and upon agroinoculation with MYMV they exhibited a severe reduction in the accumulation of MYMV DNA. Thus, the MYMV AC4 hpRNA gene has emerged as a good candidate to engineer resistance against MYMV in susceptible plants.

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Acknowledgments

We thank Prof. Thomas Hohn, University of Basel for providing Nicotiana benthamiana 16c seeds. We thank Dr. Donna Bond, University of Cambridge for her comments on the manuscript. We are thankful to Dr. K. Dharmalingam, Madurai Kamaraj University for permitting us to use the Liquid Scintillation Counter. Ms. Jasvinder Kaur and Ms. K. Bhagyalakshmi are acknowledged for the construction of pCAM-AC4-gus. This study was financially supported by the Department of Biotechnology (Project Number-BT/PR9823/AGR/36/10/2007 dt 7/12/2007), Government of India and University Grants Commission, Government of India. S.S. acknowledges the Council of Scientific and Industrial Research, Government of India for the research fellowship.

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  1. Department of Plant Biotechnology, School of Biotechnology, Madurai Kamaraj University, Madurai, 625021, India

    Sukumaran Sunitha, Gnanasekaran Shanmugapriya, Veluthambi Balamani & Karuppannan Veluthambi

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  1. Sukumaran Sunitha
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  2. Gnanasekaran Shanmugapriya
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Sunitha, S., Shanmugapriya, G., Balamani, V. et al. Mungbean yellow mosaic virus (MYMV) AC4 suppresses post-transcriptional gene silencing and an AC4 hairpin RNA gene reduces MYMV DNA accumulation in transgenic tobacco. Virus Genes 46, 496–504 (2013). https://doi.org/10.1007/s11262-013-0889-z

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