苏云金芽孢杆菌埃及分离物营养杀虫蛋白(VIPs)对白蝇毒效的生物学和分子研究

L. El-Gaied, H. El-Sheshtawy, W. Elmenofy
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引用次数: 2

摘要

烟粉虱(Gennadius)传播的双病毒在蔬菜和纤维作物中流行。它可以感染600多种寄主植物。它还被认为是传播番茄黄卷叶病毒(TYLCV)等多种双病毒的主要载体,对番茄作物造成严重危害。已有研究报道苏云金芽孢杆菌(Bacillus thuringiensis, Bt)的营养杀虫蛋白(VIPs)对不同昆虫具有杀虫活性。本研究对苏云金芽孢杆菌本地分离株BtC-18和BtDI-29进行了vip对粉虱种群的杀虫活性筛选。中位致死浓度(LC50)分析表明,苏云金芽孢杆菌BtC-18对白蝇的效价和毒性均高于BtDI-29菌株,其LC50分别为90 ppm和160 ppm。但中位致死时间(LT50)值在两株间无显著差异。对vip基因的PCR分析证实,BtC-18基因组中存在vip1、vip2和vip3基因。BtC-18培养球的蛋白提取物经80%饱和硫酸铵沉淀进一步纯化。纯化后的蛋白在88 KDa处显示一条清晰的条带,与先前证明的Vip3A蛋白相对应。纯化条带在88KDa下的LC50值为898 ppm,显示出对白蝇的杀虫活性。
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BIOLOGICAL AND MOLECULAR STUDIES ON THE TOXIC EF-FECT OF VEGETATIVE INSECTICIDAL PROTEIN (VIPs) OF Ba-cillus thuringiensis EGYPTIAN ISOLATES AGAINST WHITEFLIES
Whitefly Bemisia tabaci (Gennadius) transmitted geminiviruses cause epidemics in vegetable and fiber crops. It can infect more than 600 species of host plants. It also considered as the major vector transmitting various types of geminiviruses such as tomato yellow leaf curl virus (TYLCV) which cause great damage to tomatoes crop. Previous studies have reported that the vegetative insecticidal proteins (VIPs) of Bacillus thuringiensis (Bt) revealed insecticidal activity against different insect species. In this study, two local isolates of B. thuringiensis named BtC-18 and BtDI-29 were screened for the insecticidal activity of VIPs against whitefly population. Analysis of median lethal concentration (LC50) revealed that B. thuringiensis strain BtC-18 is more potent and toxic than BtDI-29 strain against whiteflies with an estimated LC50 of 90 ppm and 160 ppm for BtC-18 and BtDI-29, respectively. However, the median lethal time (LT50) value did not show significant difference between both isolates. PCR analysis of vip genes confirmed the presence of vip1, vip2 and vip3 genes on BtC-18 genome. Proteins extract from the BtC-18 culture pellet were further purified by 80% saturation of ammonium sulfate precipitation. The purified protein showed a clear band at 88 KDa corresponding to Vip3A protein as previously demonstrated. The LC50 of the purified band at 88KDa showed insecticidal activity against white fly with an estimated LC50 of 898 ppm.
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