Worrall Elizabeth A, Hayward Alice C, Fletcher Stephen J, Mitter Neena
Centre of Horticultural Science, Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, St Lucia, Brisbane, QLD, 4072, Australia.
Arch Virol. 2019 Jan;164(1):181-194. doi: 10.1007/s00705-018-4065-6. Epub 2018 Oct 9.
Australian bean common mosaic virus (BCMV) isolates were sequenced, and the sequences were compared to global BCMV and bean common mosaic necrosis virus (BCMNV) sequences and analysed for conserved potyviral motifs to generate in planta RNA-interference (RNAi) resistance. Thirty-nine out of 40 previously reported potyvirus motifs were conserved among all 77 BCMV/BCMNV sequences. Two RNAi target regions were selected for dsRNA construct design, covering 920 bp of the nuclease inclusion b (NIb) protein and 461 bp of the coat protein (CP). In silico prediction of the effectiveness of these constructs for broad-spectrum defence against the 77 BCMV and BCMNV sequences was done via analysis of putative 21-nucleotide (nt) and 22-nt small-interfering RNAs (siRNAs) generated from the target regions. The effectiveness of both constructs for siRNA generation and BCMV RNAi-mediated resistance was validated in Nicotiana benthamiana transient assays.
对澳大利亚菜豆普通花叶病毒（BCMV）分离株进行了测序，并将这些序列与全球BCMV和菜豆普通花叶坏死病毒（BCMNV）序列进行比较，分析其保守的马铃薯Y病毒基序，以产生植物体内RNA干扰（RNAi）抗性。在所有77个BCMV/BCMNV序列中，40个先前报道的马铃薯Y病毒基序中有39个是保守的。选择了两个RNAi靶区域用于设计双链RNA（dsRNA）构建体，覆盖核酸酶包含体b（NIb）蛋白的920 bp和外壳蛋白（CP）的461 bp。通过分析从靶区域产生的假定21核苷酸（nt）和22 nt小干扰RNA（siRNA），对这些构建体针对77个BCMV和BCMNV序列进行广谱防御的有效性进行了计算机模拟预测。在本氏烟草瞬时试验中验证了这两种构建体产生siRNA和BCMV RNAi介导抗性的有效性。
