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病毒衍生的发夹 RNA 转基因赋予烟草花叶病毒和黄瓜花叶病毒的免疫性。

Virus-derived transgenes expressing hairpin RNA give immunity to Tobacco mosaic virus and Cucumber mosaic virus.

机构信息

State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou 310029, P.R. China.

出版信息

Virol J. 2011 Jan 27;8:41. doi: 10.1186/1743-422X-8-41.

DOI:10.1186/1743-422X-8-41
PMID:21269519
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3038950/
Abstract

BACKGROUND

An effective method for obtaining resistant transgenic plants is to induce RNA silencing by expressing virus-derived dsRNA in plants and this method has been successfully implemented for the generation of different plant lines resistant to many plant viruses.

RESULTS

Inverted repeats of the partial Tobacco mosaic virus (TMV) movement protein (MP) gene and the partial Cucumber mosaic virus (CMV) replication protein (Rep) gene were introduced into the plant expression vector and the recombinant plasmids were transformed into Agrobacterium tumefaciens. Agrobacterium-mediated transformation was carried out and three transgenic tobacco lines (MP16-17-3, MP16-17-29 and MP16-17-58) immune to TMV infection and three transgenic tobacco lines (Rep15-1-1, Rep15-1-7 and Rep15-1-32) immune to CMV infection were obtained. Virus inoculation assays showed that the resistance of these transgenic plants could inherit and keep stable in T₄ progeny. The low temperature (15 °C did not influence the resistance of transgenic plants. There was no significant correlation between the resistance and the copy number of the transgene. CMV infection could not break the resistance to TMV in the transgenic tobacco plants expressing TMV hairpin MP RNA.

CONCLUSIONS

We have demonstrated that transgenic tobacco plants expressed partial TMV movement gene and partial CMV replicase gene in the form of an intermolecular intron-hairpin RNA exhibited complete resistance to TMV or CMV infection.

摘要

背景

通过在植物中表达病毒衍生的 dsRNA 诱导 RNA 沉默是获得抗性转基因植物的有效方法，该方法已成功应用于生成对许多植物病毒具有抗性的不同植物系。

结果

将烟草花叶病毒（TMV）运动蛋白（MP）基因和黄瓜花叶病毒（CMV）复制蛋白（Rep）基因的部分反向重复序列导入植物表达载体，并将重组质粒转化为根癌农杆菌。进行根癌农杆菌介导的转化，获得了对 TMV 感染免疫的 3 个转基因烟草品系（MP16-17-3、MP16-17-29 和 MP16-17-58）和对 CMV 感染免疫的 3 个转基因烟草品系（Rep15-1-1、Rep15-1-7 和 Rep15-1-32）。病毒接种试验表明，这些转基因植物的抗性可在 T₄代中遗传并保持稳定。低温（15°C）不影响转基因植物的抗性。抗性与转基因拷贝数之间没有显著相关性。表达 TMV 发夹 MP RNA 的转基因烟草植物中 CMV 感染不能破坏对 TMV 的抗性。

结论

我们已经证明，以分子间内含子-发夹 RNA 形式表达部分 TMV 运动基因和部分 CMV 复制酶基因的转基因烟草植物对 TMV 或 CMV 感染表现出完全抗性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/15c8/3038950/62eb4ee04aef/1743-422X-8-41-6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/15c8/3038950/5194df0921b4/1743-422X-8-41-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/15c8/3038950/5ddfb5ba3d84/1743-422X-8-41-2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/15c8/3038950/b468dded20c4/1743-422X-8-41-3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/15c8/3038950/8842a349d33f/1743-422X-8-41-4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/15c8/3038950/05e8a30351d7/1743-422X-8-41-5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/15c8/3038950/62eb4ee04aef/1743-422X-8-41-6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/15c8/3038950/5194df0921b4/1743-422X-8-41-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/15c8/3038950/5ddfb5ba3d84/1743-422X-8-41-2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/15c8/3038950/b468dded20c4/1743-422X-8-41-3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/15c8/3038950/8842a349d33f/1743-422X-8-41-4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/15c8/3038950/05e8a30351d7/1743-422X-8-41-5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/15c8/3038950/62eb4ee04aef/1743-422X-8-41-6.jpg

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