Crude extracts of bacterially expressed dsRNA can be used to protect plants against virus infections.
Crude extracts of bacterially expressed dsRNA can be used to protect plants against virus infections.
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DOI：
10.1186/1472-6750-3-3
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发表时间：
2003-03-20
影响因子：
3.5
通讯作者：
Díaz-Ruíz JR
中科院分区：
文献类型：
--
作者：
Tenllado F;Martínez-García B;Vargas M;Díaz-Ruíz JR
双链RNA（dsRNA）是植物、秀丽隐杆线虫、果蝇和哺乳动物等多种生物体中基因沉默的有效启动子。我们之前已经证明并获得专利，来自病毒序列的体外转录的dsRNA的机械接种特异性地防止植物中的病毒感染。该方法需要在体外合成大量的RNA，涉及高成本和相当大的劳动力。我们已经开发了一种在细菌中产生大量病毒衍生的dsRNA的体内表达系统，以期提供对植物中病毒疾病的实际控制。部分纯化的细菌双链RNA促进了植物中两种病毒感染的特异性干扰，这两种病毒属于马铃薯Y病毒属和马铃薯Y病毒属。此外，我们已经证明，当通过简单的程序喷洒到植物表面上时，容易获得的细菌表达的dsRNA的粗提取物同样有效地保护植物免受病毒感染。在病毒接种前几天用法国压榨裂解物喷洒植物时，病毒感染性显著消除。我们的方法提供了一种替代的植物物种的遗传转化与dsRNA表达的构建体能够干扰植物病毒。与再生转基因植物的要求相比，这种dsRNA生产模式的主要优点是其简单性和极低的成本。该方法不仅为植物抗病毒病提供了可靠的工具，而且为植物病毒感染中基因沉默机制的研究提供了潜在的工具。
Double-stranded RNA (dsRNA) is a potent initiator of gene silencing in a diverse group of organisms that includes plants, Caenorhabditis elegans, Drosophila and mammals. We have previously shown and patented that mechanical inoculation of in vitro-transcribed dsRNA derived from viral sequences specifically prevents virus infection in plants. The approach required the in vitro synthesis of large amounts of RNA involving high cost and considerable labour. We have developed an in vivo expression system to produce large amounts of virus-derived dsRNAs in bacteria, with a view to providing a practical control of virus diseases in plants. Partially purified bacterial dsRNAs promoted specific interference with the infection in plants by two viruses belonging to the tobamovirus and potyvirus groups. Furthermore, we have demonstrated that easy to obtain, crude extracts of bacterially expressed dsRNAs are equally effective protecting plants against virus infections when sprayed onto plant surfaces by a simple procedure. Virus infectivity was significantly abolished when plants were sprayed with French Press lysates several days before virus inoculation. Our approach provides an alternative to genetic transformation of plant species with dsRNA-expressing constructs capable to interfere with plant viruses. The main advantage of this mode of dsRNA production is its simplicity and its extremely low cost compared with the requirements for regenerating transgenic plants. This approach provides a reliable and potential tool, not only for plant protection against virus diseases, but also for the study of gene silencing mechanisms in plant virus infections.
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影响因子：
2.7
作者：
LOPEZMOYA, JJ;CANTO, T;DIAZRUIZ, JR
通讯作者：
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影响因子：
56.9
作者：
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通讯作者：
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影响因子：
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作者：
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作者：
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通讯作者：
Tsagris, M
DOI：
10.1073/pnas.200217597
发表时间：
2000-10-10
影响因子：
11.1
作者：
Fagard, M;Boutet, S;Vaucheret, H
通讯作者：
Vaucheret, H