Roles of Nicotiana tobacum NtWRKY40 in Plant Responding to Virus Infection

doi:10.13560/j.cnki.biotech.bull.1985.2016.10.023

Abstract

Abstract: Some of WRKY transcription factors have involved in plant abiotic defense. A WRKY gene,a homologue of Arabidopsis WRKY40,was found being dramatically induced by plant virus and further characterized in this study. To verify its function,NtWRKY40over-expressed tobacco plants were established. After virus infection,the transcription levels of NtWRKY40 and virus related genes were quantified using quantitative real time PCR at three stages（6 h,3 d,9 d after infection）. Results showed that tobacco mosaic virus infection led to 150 fold higher of NtWRKY40 transcript in non-transgenic wild type tobacco than non-infected control. No significant change in NtWRKY40 transcription levels was noted in the NtWRKY40 over-expressed tobacco after virus infection.NtWRKY40 over-expressed tobaccos were more sensitive to TMV infection than non-transgenic wild type. The data suggested that NtWRKY40 was a negative regulator in responding to TMV. In addition,an artificial miRNA vector amiR167-PVY was constructed,for silencing the coat protein gene of Potato virus Y,using the miR167 precursor of Nicotiana tabacum as backbone and cauliflower mosaic virus 35S promoter. Over-expression of amiR167-PVY exhibited inhibitory effect on the virus. The results provide the basis for the further research on amiRNA-mediated virus resistance mechanism,and contributes to antiviral transgenic plants cultivation with the optimal target sequence.

Key words: Tobacco mosaic virus, Potato virus Y, WRKY transcription factor, artificial microRNA, virus resistance

LIU Jing-jing, CHENG Chun-ling, XI Yu-zhen, WEI Shu. Roles of Nicotiana tobacum NtWRKY40 in Plant Responding to Virus Infection[J]. Biotechnology Bulletin, 2016, 32(10): 188-198.

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