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

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...

AI Technical Summary

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 main means. Chemical pesticides have low control efficiency, many residues on crops, harmful to humans and animals, and pollute the environment. Although the pesticide preparations currently used to prevent and control viral diseases There are many, but the real active ingredients and dosage forms are very limited; the real application of biological control in production is still very little

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