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伯氏致病杆菌(Xenor habdus bovienii BE)杀虫活性物质的分离和生物测定
引用本文:段秀峰,林茂松,李红梅,王暄,刘维红.伯氏致病杆菌(Xenor habdus bovienii BE)杀虫活性物质的分离和生物测定[J].江苏农业学报,2009,25(1).
作者姓名:段秀峰  林茂松  李红梅  王暄  刘维红
作者单位:南京农业大学植物病理学系,江苏,南京,210095
基金项目:高等学校博士学科点专项科研基金 
摘    要:对斯氏线虫(Steinernera feltiae)比利时品系的共生菌伯氏致病杆菌(Xenorhabdus bovienii BE)进行发酵培养,测定结果显示,该菌发酵液对供试昆虫大蜡螟老龄幼虫有很高的血腔毒性.对发酵液硫酸铵盐析和乙酸乙酯抽提物等衍生物进行测定,发现共生菌的杀虫物质主要为胞外和胞内蛋白成分,且胞内粗提蛋白活性明显高于胞外蛋白.对两种粗提蛋白进行Marker分子量范围为(1.43~9.72)×104 的SDS-PAGE分析,发现胞外粗提蛋白在此分子量范围内没有明显的条带,而胞内粗提蛋白则有较明显的5个条带.研究表明胞内粗提蛋白具有较高的热稳定性,测定的盐析最佳硫酸铵饱和度为80%,此粗提蛋白在24 h和48 h后对昆虫血腔毒性的LD50值分别为4.731 6 g/L和 1.571 5 g/L;其在5 d后对二龄幼虫的口服LD50为 253.757 9 μg/g.胞内粗提蛋白凝胶过滤后呈现4个峰值,分别对其进行SDS-PAGE分析,只有浓度较大的峰值1和4产生明显条带,分子量分别为6.64×104 和(2.01~2.90)×104 的2种复合蛋白;分别对它们进行血腔和口服毒性生物活性测定,发现分子量为(2.01~2.90)×104的2种复合蛋白杀虫活性高于分子量为6.64×104 蛋白的杀虫活性,其在24 h和48 h后对昆虫血腔毒性的LD50分别为 0.377 2 g/L和 0.123 8 g/L,其在5 d后对二龄幼虫的口服LD50为 93.002 1 μg/g.

关 键 词:斯氏线虫  大蜡螟  伯氏致病杆菌  血腔毒性  口服毒性

Isolation and Bioassays of Insecticidal Substances from Symbiotic Bacterium Xenorhabdus bovienii BE of Entomopathogenic Nematode Steinernera feltiae
DUAN Xiu-feng,LIN Mao-song,LI Hong-mei,WANG Xuan,LIU Wei-hong.Isolation and Bioassays of Insecticidal Substances from Symbiotic Bacterium Xenorhabdus bovienii BE of Entomopathogenic Nematode Steinernera feltiae[J].Jiangsu Journal of Agricultural Sciences,2009,25(1).
Authors:DUAN Xiu-feng  LIN Mao-song  LI Hong-mei  WANG Xuan  LIU Wei-hong
Institution:Department of Plant Pathology;Nanjing Agricultural University;Nanjing 210095;China
Abstract:The symbiotic bacterium Xenorhabdus bovienii BE strain was extracted from the Belgian population of entomopathogenic nematode Steinernera feltiae.High toxicity to Galleria mellonella fifth instar larvae was observed when the insect hemocoel cavities were injected with the fermentation solutions of the bacteria.Studies on the insecticidal activity of substances from fermentation solution indicated that the toxic substances were proteins existed in both extracellular and intracellular environment,and the toxi...
Keywords:SDS-PAGE
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