苏云金芽孢杆菌及vip基因在不同自然环境中分布情况的比较

发布时间：2018-04-21 03:14

本文选题：苏云金芽孢杆菌 + vip基因　；参考：《江苏农业科学》2017年04期

【摘要】：营养期杀虫蛋白(vegetative insecticidal protein,简称VIP)是苏云金芽孢杆菌(Bacillus thuringiensis,Bt)在营养期表达的一类新型杀虫毒蛋白,该蛋白的发现使得Bt杀虫剂在杀虫活性、杀虫范围方面得到很大突破。首次比较我国不同气候类型(热带:海南省,亚热带:广西壮族自治区,温带:黑龙江省)的Bt菌株和vip基因的分布情况。结果表明,从海南省841份土样中分离出156株Bt菌(18.55%),从广西地区1 420份土样中分离出356株Bt菌(25.07%),从黑龙江省1 010份土样中分离出167株Bt菌(16.53%);从海南省的156株Bt菌中鉴定出22个vip基因(占14.1%),从广西地区的356株Bt菌中鉴定出2个vip基因(占5.62%),然而,从黑龙江省的156株Bt菌中没有鉴定出vip基因(0%)。利用聚合酶链式反应(polymerase chain reaction,简称PCR)方法成功克隆出1个vip3Aa的全长基因并获得登录号KT792883,将该基因插入到表达载体p ET-21b中,转化菌株Rosetta(DE3),在低温条件下表达88 ku的蛋白,将该蛋白进行生物活性测定,结果显示,该蛋白对甜菜夜蛾(Spodoptera exigua)具有较好的杀虫活性,LC50为5.124 ng/mg,对棉铃虫(Heliothis armigera)活性较低,LC50为870.1 ng/mg。研究结果显示,Bt菌株和vip基因的分布有地域性差异,在温度较高的热带、亚热带地区有比较丰富的Bt菌株和vip基因资源。研究将有利于充分利用土壤中的Bt菌株,分离具有自主知识产权和重要应用价值的新型vip抗虫基因,这将会为研究新型杀虫蛋白、延缓害虫抗性问题提供有益帮助。
[Abstract]:Vegetative insecticidal protein (vip) is a new type of insecticidal protein expressed by Bacillus thuringiensis Bt. during the nutritive period. The discovery of this protein has made a great breakthrough in insecticidal activity and scope of BT insecticide. It is the first time to compare the distribution of BT strain and vip gene in different climatic types of China (tropical: Hainan province, subtropical region: Guangxi Zhuang Autonomous region, temperate zone: Heilongjiang Province). The results show that 156 strains of BT bacteria were isolated from 841 soil samples in Hainan Province, 356 strains of BT bacteria were isolated from 1,420 soil samples from Guangxi region, 167 strains of BT bacteria were isolated from 1,010 soil samples in Heilongjiang Province, and 22 strains were identified from 156 strains of BT bacteria in Hainan Province. Two vip genes (5.62%) were identified from 356 BT strains in Guangxi. The vip gene was not identified from 156 BT strains in Heilongjiang Province. A full-length vip3Aa gene was successfully cloned by polymerase chain reaction (PCR) method, and the accession number KT792883 was obtained. The gene was inserted into the expression vector p ET-21b. The transformed strain Rosetta-DE3H expressed 88 ku protein at low temperature. The bioactivity of the protein was determined. The results showed that the protein had a better insecticidal activity against Spodoptera exigua (5.124 ng / mg) and a lower activity of 870.1 ng / mgagainst Helicoverpa armigera (Heliothis armigera). The results showed that there were regional differences in the distribution of BT and vip genes, and there were abundant BT strains and vip gene resources in the tropics and subtropics where the temperature was higher. The study will be beneficial to the utilization of BT strains in soil and the isolation of new vip insecticidal genes with independent intellectual property rights and important application value, which will provide useful help for studying new insecticidal proteins and delaying the problem of pest resistance.
【作者单位】：东北农业大学;
【基金】：国家自然科学基金(编号:31401812) 东北农业大学科学研究基金(编号:2010RCB54)
【分类号】：S476

【相似文献】