| abstract |
The invention provides a tobacco anti-virus regulatory gene screening method and application. The method firstly uses a virus-induced gene silencing VIGS vector as a target vector to construct a tobacco cDNA library, which can be directly used for gene silencing screening and analysis after homogenization treatment. The VIGS-cDNA library was obtained by transforming the VIGS-cDNA library plasmid into Agrobacterium to obtain an Agrobacterium library; the Agrobacterium library was plated and cultured to obtain a single colony, and each single colony was subjected to VIGS silencing analysis, and the antivirus-related colonies were obtained by screening. ; Sequencing and analyzing the genes corresponding to the anti-virus-related colonies obtained by screening, and performing homology analysis and function prediction on the obtained sequences to obtain anti-viral regulatory genes. The method of the invention is easy to operate, does not need to construct transgenic plants or mutants, takes less time, and has high overall screening efficiency, is particularly suitable for screening of mutant lethal genes, and has broad application prospects. |