Position: Home > Articles > Constructing Transgenic Tobacco with Resistance against Potato Virus Y Using Synthetic Trans-siRNA
Shandong Agricultural Sciences
2018
(12)
86-90
利用人工ta-siRNA策略培育抗PVY烟草
作 者：
吴斌;姜珊珊;张眉;徐德坤;辛志梅;王升吉
单 位：
山东省农业科学院植物保护研究所;临沂市植物保护站
关键词：
人工ta-siRNA;马铃薯Y病毒(PVY);RNA介导的病毒抗性;转基因烟草
摘 要：
人工ta-siRNA基因沉默技术具有同时表达多个ta-siRNA和产生ta-siRNA位置固定等特点,在精确性和高效性等方面有明显优势。本研究以马铃薯Y病毒(potato virus Y,PVY)外壳蛋白(coat protein,CP)为靶基因,选取不同位置的3个siRNA为靶序列,烟草TAS3a作为ta-siRNA表达的骨架,分别构建植物表达载体PVY-CP5'、PVY-CPM和PVY-CP3',并利用叶盘法转化烟草NC89。瞬时表达检测结果显示,3个syn-tasiRNA植物表达载体均能在植物体内成功表达siRNA;抗病性鉴定发现,转PVY-CP5'、PVY-CPM和PVY-CP3'的转基因株系中抗病株系的比率分别为45. 08%、27. 21%、61. 90%;Northern杂交分析显示,目的片段转录产物(总RNA)的积累量与植株的抗病性呈负相关,转基因植株中均有siRNA存在,表明植株的病毒抗性是由RNA介导的。
译 名：
Constructing Transgenic Tobacco with Resistance against Potato Virus Y Using Synthetic Trans-siRNA
作 者：
Wu Bin;Jiang Shanshan;Zhang Mei;Xu Dekun;Xin Zhimei;Wang Shengji;Institute of Plant Protection,Shangdong Academy of Agricultural Sciences;Plant Protection Station of Linyi City;
单 位：
Wu Bin%Jiang Shanshan%Zhang Mei%Xu Dekun%Xin Zhimei%Wang Shengji%Institute of Plant Protection,Shangdong Academy of Agricultural Sciences%Plant Protection Station of Linyi City
关键词：
Synthetic trans-siRNA;;Potato virus Y(PVY);;RNA-mediated virus resistance;;Transgenic tobacco
摘 要：
Synthetic trans-siRNA has the characteristics of simultaneous expression of multiple ta-siRNA and fixed location,which has obvious advantages in accuracy and efficiency. In this study,synthetic trans-siRNA constructs based on 3 regions of the CP gene of PVY were designed by using TAS3 a of tobacco as ta-siRNA expression skeleton. The plant expression vectors dubbed PVY-CP5',PVY-CPM and PVY-CP3' were transformed into tobacco NC89 by leaf disc method. The siRNAs were detected in all expression vectors in transient assay,the ratio of disease-resistant lines in transgenic lines of PVY-CP5',PVY-CPM and PVY-CP3' were 45. 08%,27. 21% and 61. 90%,respectively. Northern blot analysis revealed an inverse correlation between accumulation of target transcripts(total RNA) and plant disease resistance,siRNAs were present in all selected resistant and susceptible transgenic plants. The results proved this resistance was RNA-mediated.
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