摘要：In this paper, the mrrl gene of Botrytis cinerea (B.cinerea)was cloned, and its resistance to six types of soil pollutants was tested. The plate trans-parent circle method was used to screen strains, and a strain of B.cinerea PTH-mrr1 with activity against multiple soil pollutants was obtained. Re-al-Time PCR found that mrr-I gene was signifi-cantly down-regulated in this strain. PCR technol-ogy was used to successfully clone the mrr-I mutant gene from this strain. Sequencing results revealed that the mrrI mutant gene contained 2 490 bases and 830 amino acids. Compared with the normal base sequence, multiple base insertions or deletions occurred at multiple sites. The mrr7 mutant gene was expressed in E.coli to obtain a prokaryotic ex-pression product, which was then used to act on a variety of soil pollutants. It was found that E.coli with mrrl gene mutation was resistant to car-bendazim,procymidone, iprodione, fludioxonil, pyrimethamnil and thiram.The prokaryotic expres-sion conditions were further optimized, and it was found that when E. coli containing mrrl gene was cultured with shaking at 30℃C for 24 hours,and the IPTG concentration was 0.75 mmol/L, the expres-sion level of mrrl protein was the highest. Fur-thermore, it has the best resistance to carbendazim, procymidone, iprodione,fludioxonil,pyrimethanil and thiram. In conclusion, in this study, we dis-cussed the gene function of B.cinerea mrrl and determined its resistance to various soil pollutants. This research laid a theoretical foundation for the future development ofantisoil pollutant products.
关键词：Bcinerea;mrl gene;multiple site mutation;soil pollu-tant;resistance test
