Kung Yi-Jung, You Bang-Jau, Raja Joseph A J, Chen Kuan-Chun, Huang Chiung-Huei, Bau Huey-Jiunn, Yang Ching-Fu, Huang Chung-Hao, Chang Chung-Ping, Yeh Shyi-Dong
1] Department of Plant Pathology, National Chung Hsing University, Taichung, Taiwan, R.O.C. [2] NCHU-UCD Plant and Food Biotechnology Center, National Chung Hsing University, Taiwan, R.O.C.
1] Department of Plant Pathology, National Chung Hsing University, Taichung, Taiwan, R.O.C. [2] Department of Chinese Pharmaceutical Sciences and Chinese Medicine Resources, China Medical University, Taichung, Taiwan, R.O.C.
Sci Rep. 2015 Apr 27;5:9804. doi: 10.1038/srep09804.
Controlling plant viruses by genetic engineering, including the globally important Papaya ringspot virus (PRSV), mainly involves coat protein (CP) gene mediated resistance via post-transcriptional gene silencing (PTGS). However, the breakdown of single- or double-virus resistance in CP-gene-transgenic papaya by more virulent PRSV strains has been noted in repeated field trials. Recombination analysis revealed that the gene silencing suppressor HC-Pro or CP of the virulent PRSV strain 5-19 is responsible for overcoming CP-transgenic resistance in a sequence-homology-independent manner. Transient expression assays using agro-infiltration in Nicotiana benthamiana plants indicated that 5-19 HC-Pro exhibits stronger PTGS suppression than the transgene donor strain. To disarm the suppressor from the virulent strain, transgenic papaya lines were generated carrying untranslatable 5-19 HC-Pro, which conferred complete resistance to 5-19 and other geographic PRSV strains. Our study suggested the potential risk of the emergence of more virulent virus strains, spurred by the deployment of CP-gene-transgenic crops, and provides a strategy to combat such strains.
通过基因工程控制植物病毒,包括全球重要的番木瓜环斑病毒(PRSV),主要涉及通过转录后基因沉默(PTGS)介导的外壳蛋白(CP)基因抗性。然而,在反复的田间试验中发现,更具毒性的PRSV毒株会导致CP基因转基因番木瓜中的单病毒或双病毒抗性失效。重组分析表明,毒性PRSV毒株5-19的基因沉默抑制因子HC-Pro或CP以序列同源性无关的方式克服了CP转基因抗性。在本氏烟草植株中使用农杆菌浸润进行的瞬时表达试验表明,5-19 HC-Pro比转基因供体毒株表现出更强的PTGS抑制作用。为了去除毒性毒株中的抑制因子,构建了携带不可翻译的5-19 HC-Pro的转基因番木瓜品系,该品系对5-19及其他地理区域的PRSV毒株具有完全抗性。我们的研究表明,CP基因转基因作物的应用可能会促使出现更具毒性的病毒毒株,并提供了应对此类毒株的策略。