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利用 RNAi 介导的方法来抵抗转基因木薯中的乌干达木薯褐色条斑病毒。

RNAi-mediated resistance to Cassava brown streak Uganda virus in transgenic cassava.

机构信息

International Laboratory for Tropical Agricultural Biotechnology, Donald Danforth Plant Science Center, St. Louis, MO 63132, USA.

出版信息

Mol Plant Pathol. 2011 Sep;12(7):677-87. doi: 10.1111/j.1364-3703.2010.00700.x. Epub 2011 Feb 17.

Abstract

Cassava brown streak disease (CBSD), caused by Cassava brown streak Uganda virus (CBSUV) and Cassava brown streak virus (CBSV), is of new epidemic importance to cassava (Manihot esculenta Crantz) production in East Africa, and an emerging threat to the crop in Central and West Africa. This study demonstrates that at least one of these two ipomoviruses, CBSUV, can be efficiently controlled using RNA interference (RNAi) technology in cassava. An RNAi construct targeting the near full-length coat protein (FL-CP) of CBSUV was expressed constitutively as a hairpin construct in cassava. Transgenic cassava lines expressing small interfering RNAs (siRNAs) against this sequence showed 100% resistance to CBSUV across replicated graft inoculation experiments. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis showed the presence of CBSUV in leaves and some tuberous roots from challenged controls, but not in the same tissues from transgenic plants. This is the first demonstration of RNAi-mediated resistance to the ipomovirus CBSUV in cassava.

摘要

木薯褐色条斑病(CBSD)由乌干达木薯褐色条斑病毒(CBSUV)和木薯褐色条斑病毒(CBSV)引起,对东非的木薯(Manihot esculenta Crantz)生产具有新的流行重要性,也是中、西非该作物的一种新出现的威胁。本研究表明,至少有一种杆状病毒,即 CBSUV,可以通过 RNA 干扰(RNAi)技术在木薯中得到有效控制。针对 CBSUV 近全长外壳蛋白(FL-CP)的 RNAi 构建体作为发夹结构在木薯中组成型表达。表达针对该序列的小干扰 RNA(siRNA)的转基因木薯系在重复的嫁接接种实验中表现出对 CBSUV 的 100%抗性。逆转录聚合酶链反应(RT-PCR)分析显示, challenged 对照的叶片和一些块根中存在 CBSUV,但转基因植物的相同组织中不存在。这是 RNAi 介导的对木薯 CBSUV 的杆状病毒抗性的首次证明。

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