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Analysis of transitive RNA silencing after grafting in transgenic plants with the coat protein gene of Sweet potato feathery mottle virus

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Abstract

We have previously reported the graft transmission of target specificity for RNA silencing using transgenic Nicotiana benthamiana plants expressing the coat protein gene (CP, including the 3′ non-translated region) of Sweet potato feathery mottle virus. Transgenic plants carrying the 5′ 200 and 400 bp regions of CP were newly produced. From these plants, two silenced and two non-silenced lines were selected to investigate the manifestation of transitive RNA silencing by graft experiments. Non-silenced scions carrying the entire transgene were grafted onto either 5′ or 3′ silencing inducer rootstocks. When non-silenced scions were grafted onto 5′ silencing inducer rootstocks, RNA silencing was induced in the non-silenced scions and spread toward the 3′ region of the transgene mRNA. Similarly, when non-silenced scions were grafted onto 3′ silencing inducer rootstocks, RNA silencing was induced in the non-silenced scions, but was restricted to the 3′ region of the transgene and did not spread to the 5′ region. In addition, results from crossing experiments, involving non-silenced and 3′ silencing inducer plants, confirmed the above finding. This indicates that RNA silencing spreads in the 5′–3′ direction, not in the 3′–5′ direction, along the transgene mRNA.

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Abbreviations

abRNA:

Aberrant RNA

Ago:

Argonaute

CP :

Coat protein gene

dsRNA:

Double-stranded RNA

IR:

Inverted repeat

PTGS:

Post-transcriptional gene silencing

PVX:

Potato virus X

RdRP:

RNA-dependent RNA polymerase

RISC:

RNA-induced silencing complex

siRNA:

Short interfering RNA

SPFMV:

Sweet potato feathery mottle virus

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Acknowledgements

The authors would like to thank David Baulcombe (The Sainsbury Laboratory, Norwich, UK) and his other lab members for kindly providing the PVX vector and specific comments on the crossing of N. benthamiana. They are also grateful to Sk. Md. Fazle Akbar (School of Medicine, Ehime University, Japan) and Thangavelu U. Arumugam (Faculty of Agriculture, Ehime University) for critical reading of the manuscript, and Charles Lecellier (Institut de Biologie Moleculaire des Plantes - CNRS UPR2357, France) for suggestions concerning the siRNA detection experiment. This work was financially supported in part by Ministry of Education, Culture, Sports, Science and Technology of Japan and by Ministry of Agriculture, Forestry and Fisheries of Japan.

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Authors and Affiliations

  1. Faculty of Agriculture, Ehime University, 3-5-7 Tarumi, Matsuyama, Ehime, 790-8566, Japan

    A. K. M. N. Haque & Masamichi Nishiguchi

  2. National Institute of Agrobiological Resources, Kan-nondai, Tsukuba, Ibaraki, 305-8602, Japan

    Yoshikazu Tanaka, Shoji Sonoda & Masamichi Nishiguchi

  3. Wakasa Wan Energy Research Center, Tsuruga, Fukui, 914-0192, Japan

    Yoshikazu Tanaka

  4. Research Institute of Bioresources, Okayama University, Kurashiki, Okayama, 710-0046, Japan

    Shoji Sonoda

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  1. A. K. M. N. Haque
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Correspondence to Masamichi Nishiguchi.

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Haque, A.K.M.N., Tanaka, Y., Sonoda, S. et al. Analysis of transitive RNA silencing after grafting in transgenic plants with the coat protein gene of Sweet potato feathery mottle virus . Plant Mol Biol 63, 35–47 (2007). https://doi.org/10.1007/s11103-006-9070-6

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