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Untranslatable tospoviral NSs fragment coupled with L conserved region enhances transgenic resistance against the homologous virus and a serologically unrelated tospovirus

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Abstract

Tospoviruses cause severe damages to important crops worldwide. In this study, Nicotiana benthamiana transgenic lines carrying individual untranslatable constructs comprised of the conserved region of the L gene (denoted as L), the 5′ half of NSs coding sequence (NSs) or the antisense fragment of whole N coding sequence (N) of Watermelon silver mottle virus (WSMoV), individually or in combination, were generated. A total of 15–17 transgenic N. benthamiana lines carrying individual transgenes were evaluated against WSMoV and the serologically unrelated Tomato spotted wilt virus (TSWV). Among lines carrying single or chimeric transgenes, the level of resistance ranged from susceptible to completely resistant against WSMoV. From the lines carrying individual transgenes and highly resistant to WSMoV (56–63 % of lines assayed), 30 % of the L lines (3/10 lines assayed) and 11 % of NSs lines (1/9 lines assayed) were highly resistant against TSWV. The chimeric transgenes provided higher degrees of resistance against WSMoV (80–88 %), and the NSs fragment showed an additive effect to enhance the resistance to TSWV. Particularly, the chimeric transgenes with the triple combination of fragments, namely L/NSs/N or HpL/NSs/N (a hairpin construct), provided a higher degree of resistance (both 50 %, with 7/14 lines assayed) against TSWV. Our results indicate that the untranslatable NSs fragment is able to enhance the transgenic resistance conferred by the L conserved region. The better performance of L/NSs/N and HpL/NSs/N in transgenic N. benthamiana lines suggests their potential usefulness in generating high levels of enhanced transgenic resistance against serologically unrelated tospoviruses in agronomic crops.

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Acknowledgments

We thank the funding supports of the Ministry of Science and Technology (NSC92-2317-B-005-024, NSC-101-2911-I-005-301 and NSC-102-2911-I-005-301), and the Ministry of Education, Taiwan, ROC under the ATU plan for this study.

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No potential conflicts of interest are disclosed.

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Authors and Affiliations

  1. Department of Plant Pathology, College of Agriculture and Natural Resource, National Chung Hsing University, 250-Kuo-Kuang Road, Taichung, 40227, Taiwan, ROC

    Uthaman Yazhisai, Prem Anand Rajagopalan & Shyi-Dong Yeh

  2. Agricultural Biotechnology Center, National Chung Hsing University, Taichung, 40227, Taiwan, ROC

    Uthaman Yazhisai & Shyi-Dong Yeh

  3. NCHU-UCD Plant and Food Biotechnology Center, National Chung Hsing University, 250-Kuo-Kuang Road, Taichung, 40227, Taiwan, ROC

    Joseph A. J. Raja

  4. Department of Biotechnology, Asia University, Wufeng, Taichung, 41354, Taiwan, ROC

    Tsung-Chi Chen

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  1. Uthaman Yazhisai
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  2. Prem Anand Rajagopalan
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  3. Joseph A. J. Raja
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  4. Tsung-Chi Chen
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  5. Shyi-Dong Yeh
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Correspondence to Shyi-Dong Yeh.

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Yazhisai, U., Rajagopalan, P.A., Raja, J.A.J. et al. Untranslatable tospoviral NSs fragment coupled with L conserved region enhances transgenic resistance against the homologous virus and a serologically unrelated tospovirus. Transgenic Res 24, 635–649 (2015). https://doi.org/10.1007/s11248-015-9865-9

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  • DOI: https://doi.org/10.1007/s11248-015-9865-9

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