Method for cultivating tobacco capable of resisting various viruses by adopting RNAi (RNA interference) technique

An anti-virus and tobacco technology, applied in botanical equipment and methods, biochemical equipment and methods, applications, etc., can solve the problems of less biological control, low chemical pesticide control effect, limited effective ingredients and formulations, etc.

Active Publication Date: 2013-02-27
ZHENGZHOU TOBACCO RES INST OF CNTC
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Problems solved by technology

However, the breeding of disease-resistant varieties has problems such as long breeding time and prone to loss of resistance; agricultural measures can only reduce or prevent the initial source of virus infection; in terms of chemical agents, there is currently no good way to treat tobacco Viral diseases mainly take the prevention and treatment of virus transmission media as the mai...

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  • Method for cultivating tobacco capable of resisting various viruses by adopting RNAi (RNA interference) technique
  • Method for cultivating tobacco capable of resisting various viruses by adopting RNAi (RNA interference) technique
  • Method for cultivating tobacco capable of resisting various viruses by adopting RNAi (RNA interference) technique

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Embodiment Construction

[0048] The present invention will be further described below in conjunction with the accompanying drawings and implementation examples. The methods used in the following implementation examples are conventional methods unless otherwise specified.

[0049] (1) Design and synthesis of chimeric genes

[0050] By comparing the full-length genome sequences of the four viruses (CMV, PVY, PVX, TMV) in GenBank, multiple relatively conserved regions of the four viruses were screened across the entire genome. In addition, according to relevant literature reports, the 2b encoded by the CMV genome and the Hc-Pro protein encoded by PVY are RNAi suppressors (RNAi suppressor), which can inhibit the RNAi mechanism used by the host to resist the virus. In order to prevent the virus from potentially inhibiting the RNAi effect, the present invention designs an anti-CMV 2b , PVY Hc-Pro The gene selects the targeted region. TMV and PVX both selectively target CP Gene (encodes coat protein...

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Abstract

The invention relates to a method for cultivating tobacco capable of resisting various viruses by adopting an RNAi (RNA interference) technique. The method comprises the following steps of: obtaining relatively conservative areas of four kinds of viruses within a genome range through screening by comparing full-length sequences of multiple genomes of four kinds of viruses, i.e. CMV (cucumber mosaic virus), PVY (potato virus Y), PVX (potato virus X) and TMV (tobacco mosaic virus) in a genBank; selecting virus sequences and artificially synthesizing 800bp mosaic genes; and accordingly constructing RNAi plant expression carriers of the mosaic genes and transforming common tobacco through agrobacterium to obtain transgenic plants. The method for cultivating tobacco capable of resisting various viruses by adopting the RNAi technique has the characteristics that the four kinds of major tobacco viruses in China are selected as targets, the artificially constructed hairpin structures comprising the sequences of the four kinds of viruses are transformed into the tobacco by using the plant genetic engineering technique, a hairpin double-strand RNA structure transcribed by the tobacco is cut into siRNA (small interfering RNA) by the plant self mechanism, the normal duplication and the accumulation of target virus genes in tobacco plants are specifically interfered, degraded or silenced, and new tobacco materials capable of resisting various viruses can be obtained through screening.

Description

[0001] technical field [0002] The invention belongs to the technical field of plant genetic engineering, and specifically relates to a method for cultivating tobacco resistant to various viruses using RNAi technology, mainly for tobacco cucumber mosaic virus ( Cucumber mosaic virus , CMV), tobacco potato virus Y ( Potato Y virus , PVY), tobacco common mosaic virus ( Tobacco mosaic virus , TMV) and tobacco potato virus X ( Potato virus X , PVX) RNA interference vector construction method and its application in tobacco. [0003] Background technique [0004] Viral disease is a disease that cannot be ignored in tobacco production, and is an important cause of tobacco production loss. The main types of viral diseases in my country are TMV, CMV, PVY, PVX and tobacco etch disease ( Tobacco etch virus , TEV) etc. Before the 1960s, TMV was the most harmful in the tobacco-growing areas of the country; after the 1970s, the CMV hazards in the Huanghuai tobacco-growi...

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Application Information

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IPC IPC(8): C12N15/84A01H5/00
Inventor 杨军张俊祺宋纪真王燃罗朝鹏李锋魏春阳曹培健林福呈
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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