ISOLATION, TOXICITY AND MOLECULAR CHARACTERIZATION OF NATIVE Bacillus thuringiensis ISOLATES FROM EGYPTIAN SOIL

A. Makhlouf, Amira A. El Kemany, A. Riad, Y. A. Abou-Youssef
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引用次数: 2

Abstract

Thirty three local soil samples were collected from Westward till Sewa Oasis in Egypt to search novel isolates of Bacillus thuringiensis (Bt) and evaluate their toxic potentiality to overcome the serious problem of evolved resistance by insects to the pesticidal activity. The first instars larvae of cotton leaf worm (Spodoptera littoralis) were used to test their toxic potentiality in the presence of the two standard strains kurstaki (K) and neoleonensis H24a (N). The results showed that three isolates from Alexandria (AL3, AL7 and AL11) and two isolates from Kafr - el-Dawar (KD2 and KD3) were presumptively confirmed as Bt. by morphological and microscopic characters. The treated larvae with Insecticidal crystal protein (ICP) exhibited mortality percentage around 50% for all isolates except AL3 isolate. The vegetative insecticidal proteins (VIPs) of KD3, KD2 and AL11 isolates revealed mortality percentages more than 75%. While, AL3 and AL7 isolates showed differential toxicity. Crystal proteins analysis by SDS-PAGE showed KD2, KD3 and AL11 isolates gave similar profiles as those of "K" strain which was characterized with 135 kDa and 70 kDa bands. AL3 isolate failed to show any noticeable bands. According to their plasmids patterns, 4 kb was shown in all isolates and the reference strains except AL3 isolate. Cry1, Vip1 and Vip2 genes of the isolates was detected by polymerase chain reaction (PCR). The results indicated that one band with 550bp in size was present in all isolates except the AL3 isolate. Vip1 primer succeeds to amplify a band 400 bp in size in all isolates and “K" strain. Vip2 primer failed to react with any genome of the studied isolates or reference strains. This study suggested that KD2, KD3 and AL11 isolates may lead to be identified as potential strains of Bt. for their use in the development of bioinsecticide to control insect pests in Egypt.
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从埃及土壤中分离的苏云金芽孢杆菌的分离、毒性和分子特性
为了寻找苏云金芽孢杆菌(Bacillus thuringiensis, Bt)的新分离株,并评估其毒性潜力,以克服昆虫对农药活性产生进化抗性的严重问题,在埃及西田塞瓦绿洲采集了33份当地土壤样本。用棉花叶虫1龄幼虫对两种标准菌株kurstaki (K)和neoleonensis H24a (N)进行毒力测试。结果表明,来自Alexandria的3株菌株(AL3、AL7和AL11)和来自Kafr - el-Dawar的2株菌株(KD2和KD3)通过形态学和显微特征推定为Bt.。除AL3菌株外,其余菌株经杀虫晶体蛋白(ICP)处理后的幼虫死亡率均在50%左右。KD3、KD2和AL11的营养杀虫蛋白致死率均在75%以上。而AL3和AL7的毒性差异较大。SDS-PAGE晶体蛋白分析显示,KD2、KD3和AL11与“K”菌株具有相似的晶体蛋白图谱,分别具有135 kDa和70 kDa条带。AL3分离物没有显示任何明显的条带。根据质粒结构,除AL3外,其余菌株均为4 kb。采用聚合酶链反应(PCR)检测分离株Cry1、Vip1和Vip2基因。结果表明,除AL3外,其余菌株均存在一条550bp的条带。Vip1引物在所有菌株和“K”菌株中扩增出400bp的条带。Vip2引物不能与所研究的分离株或参考菌株的任何基因组发生反应。本研究提示,KD2、KD3和AL11分离株可能被鉴定为潜在的bt菌株,用于开发生物杀虫剂来防治埃及的害虫。
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TRANSFER OF GLUCANASE GENE TO RESIST LATE BLIGHT DISEASE IN POTATO (Solanum tuberosum L.) ANTIMUTAGENICS EFFECTS OF STIGMASTEROL ON TWO SALT STRESSED Lupinus termis CULTIVARS POTENTIAL EFFECT OF SOME NATURAL FOOD ADDITIVES AGAINST MONOSODIUM GLUTAMATE-INDUCED GENOTOXICITY IN Vicia faba ASSESSMENT OF GENETIC COMPONENTS AND GENETIC DIVERSITY OF SIX EGYPTIAN CLOVER (Trifolium alexandrinum L.) GENOTYPES USING ISSR AND URP MARKERS ASSOCIATION OF PNPLA3 (rs738409) AND GCKR (rs1260326) GENE POLYMORPHISMS WITH THE DEVELOPMENT OF NONALCOHOLIC FATTY LIVER DISEASE IN OBESE EGYPTIAN CHILDREN
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