發(fā)布時間：2018-05-18 07:30

  本文選題：莫海威芽胞桿菌 + GFP基因　； 參考：《植物保護學(xué)報》2017年04期

【摘要】：為明確生防莫海威芽胞桿菌Bacillus mojavensis菌株ZA1在作物體內(nèi)定殖與作物和病原物的作用機理,運用電擊轉(zhuǎn)化法將綠色熒光蛋白表達(dá)載體pGFP78導(dǎo)入菌株ZA1體內(nèi),并采用細(xì)菌培養(yǎng)法、繼代培養(yǎng)法和平板抑制法檢測其功能穩(wěn)定性。結(jié)果表明,質(zhì)粒pGFP78成功導(dǎo)入菌株ZA1,標(biāo)記后菌株ZA1-gfp經(jīng)形態(tài)學(xué)和分子生物學(xué)驗證均證明菌株ZA1已成功標(biāo)記,標(biāo)記菌株ZA1-gfp與菌株ZA1形態(tài)一致,在熒光顯微鏡下呈現(xiàn)明亮的綠色,并且能夠提取導(dǎo)入質(zhì)粒pGFP78,擴展出質(zhì)粒所攜帶的啟動子78序列。功能穩(wěn)定性鑒定結(jié)果表明,標(biāo)記菌株ZA1-gfp遺傳穩(wěn)定性達(dá)100%;與菌株ZA1的生長速率差異不顯著,且不影響菌株的運動性;菌株ZA1對馬鈴薯枯萎病菌、馬鈴薯炭疽病菌和馬鈴薯壞疽病菌的抑制率分別為64.50%、68.35%和66.55%,標(biāo)記菌株ZA1-gfp對3種病原真菌的抑制率分別為67.46%、76.56%和66.75%,表明外源質(zhì)粒的導(dǎo)入,不影響菌株的抑菌能力。
[Abstract]:In order to clarify the mechanism of colonization, crops and pathogens of Bacillus mojavensis strain ZA1 in crops, the green fluorescent protein expression vector (pGFP78) was introduced into ZA1 by electroporation, and the bacterial culture method was used. The functional stability was detected by subculture method and plate inhibition method. The results showed that the plasmid pGFP78 was successfully introduced into strain ZA1, and the labeled strain ZA1-gfp was proved by morphological and molecular biological verification to be marked successfully. The labeled strain ZA1-gfp was identical to the strain ZA1 in morphology and appeared bright green under fluorescence microscope. Moreover, the plasmid pGFP78 can be extracted and the promoter 78 sequence carried by the plasmid can be expanded. The results of functional stability identification showed that the genetic stability of the labeled strain ZA1-gfp was 100%, the growth rate of the labeled strain was not significantly different from that of strain ZA1, and it did not affect the motility of the strain. The inhibition rates of anthracnose and gangrene were 64.50% and 66.55%, respectively. The inhibition rates of the labeled strain ZA1-gfp on the three pathogenic fungi were 76.56% and 66.75%, respectively, indicating that the introduction of exogenous plasmids did not affect the bacteriostatic ability of the strains.
【作者單位】： 甘肅農(nóng)業(yè)大學(xué)植物保護學(xué)院甘肅省農(nóng)作物病蟲害生物防治工程實驗室;甘肅農(nóng)業(yè)大學(xué)甘肅省作物遺傳改良與種質(zhì)創(chuàng)新重點實驗室;
【基金】：國家現(xiàn)代農(nóng)業(yè)(馬鈴薯)產(chǎn)業(yè)技術(shù)體系(CARS-10-P18) 國家自然科學(xué)基金(31660148)
【分類號】：S476.1
，

本文編號：1904970