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Title: Exploring the trans-acting short interfering RNAs (ta-siRNAs) technology for virus control in plants

Author
item PADMANABHAN, CHELLAPPAN
item ZHENG, YI - BOYCE THOMPSON INSTITUTE
item Shamimuzzaman, Md - Shamim
item FEI, ZHANGJUN - BOYCE THOMPSON INSTITUTE
item Ling, Kai-Shu

Submitted to: American Phytopathological Society Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 5/5/2017
Publication Date: 12/1/2017
Citation: Padmanabhan, C., Zheng, Y., Shamimuzzaman, M., Fei, Z., Ling, K. 2017. Exploring the trans-acting short interfering RNAs (ta-siRNAs) technology for virus control in plants. American Phytopathological Society Annual Meeting. 107(12S):S5.91. https://doi.org/10.1094/PHYTO-107-12-S5.1.
DOI: https://doi.org/10.1094/PHYTO-107-12-S5.1

Interpretive Summary: N/A

Technical Abstract: Small ribonucleic acid (RNAs) (~20-24nt) processed from double-stranded RNA in plants can trigger degradation of the target mRNAs in cytoplasm or de novo DNA methylation in nucleus leading to gene silencing. Trans-acting short-interfering RNAs (ta-siRNAs) have been shown to enhance the target mRNA degradation and maintenance of host gene expression. In this study, we were interested in exploring ta-siRNAs technology for virus control on tomato. It was expected that by targeting the viral genomic RNA for degradation, these transgenic plants might interfere or suppress virus replication, thus confer disease resistance in the transgenic plants. Using 22 nt miRNAs targeting the viral RNA genomes of Pepino mosaic virus (PepMV) or Tomato spotted wilt virus (TSWV) for insertion into tomato pre-miRNA sequences, we were able to generate gene constructs with multiple virus-specific ta-siRNAs (21 nt). Through Agrobacterium-mediated transformation, transgenic tomato plants containing the ta-siRNAs against PepMV or TSWV were generated and the transgene integration was confirmed by polymerase chain reaction (PCR). In screening transgenic plants for virus resistance through mechanical inoculation, although no immunity to the respective virus infection was observed, a number of transgenic plants exhibited delayed or milder symptoms over control tomato plants under the same treatments. The ta-siRNA-mediated resistance may be a novel strategy in controlling viruses in plants.