Bacterially expressed dsRNA protects maize against SCMV infection.
RNA interference (RNAi) is a sequence-specific, posttranscriptional gene silencing (PTGS) process in plants that is mediated by dsRNA homologous to the silenced gene(s). In this study, we report an efficient method to produce dsRNA using a bacterial expression system. Two fragments of the Sugarcane Mosaic Virus (SCMV) CP (coat protein) gene were amplified by RT-PCR, and cloned into the inverted-repeat cloning vector pUCCRNAi. The two recombinant plasmids were transformed individually into E. coli HT115, an RNase-III deficient strain, and dsRNA was induced by isopropyl-β-D: -thiogalactopyranoside (IPTG). The crude extracts of E. coli HT115 containing large amounts of dsRNA were applied to plants as a spray and the experiment confirmed a preventative efficacy. Our findings demonstrated that spraying crude dsRNA-containing extracts inhibited SCMV infection, and the dsRNA derived from an upstream region (CP1) was more effective than was dsRNA derived from a downstream region (CP2) of the SCMV CP gene. The results provide a valuable tool for plant viral control using dsRNA and the PTGS approach.
Gan
Defang
D
Anhui Province Key Laboratory of Crop Biology, School of Life Science, Anhui Agricultural University, Hefei, 230036, China.
Zhang
Jiao
J
Jiang
Haibo
H
Jiang
Tong
T
Zhu
Suwen
S
Cheng
Beijiu
B
D036022:Capsid Proteins / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D009029:Mosaic Viruses / Q000235:genetics / Q000472:pathogenicity*; D010935:Plant Diseases / Q000517:prevention & control* / Q000821:virology; D010957:Plasmids; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000494:pharmacology*; D003313:Zea mays / Q000821:virology*
D036022:Capsid Proteins; D012330:RNA, Double-Stranded; D007544:Isopropyl Thiogalactoside