厚垣普可尼亚菌发酵液中胞外酶的活性分析及其杀虫毒力测定  

Analysis of extracellular hydrolyase activity inPochonia chlamydosporia fermentation broth and its determination of nematode killing virulence

作　　者：郝陆瑶 焦万明 尹衍峰 马园 张艳妮 李倩楠 符江涛 贾晓晴 訾星月 马俊杰 郭志凯 丁玉林[1] 杜山[1] 陆静[1] 王瑞[1] HAO Lu-yao;JIAO Wan-ming;YIN Yan-feng;MA Yuan;ZHANG Yan-ni;LI Qian-nan;FU Jiang-tao;JIA Xiao-qing;ZI Xing-yue;MA Jun-jie;GUO Zhi-kai;DING Yu-lin;DU Shan;LU Jing;WANG Rui(College of Veterinary Medicine,Key Laboratory of Clinical Diagnosis and Treatment of Animal Diseases,Ministry of Agriculture,National Animal Medicine Experimental Teaching Center,Inner Mongolia Agricultural University,Hohhot 010010,China;Animal Disease Prevention and Control Center,Ejin Banner,Alxa League 735400,China;Animal Husbandry and Veterinary Medicine Development Center of Changqing District,Jinan 250399,China)

机构地区：[1]内蒙古农业大学兽医学院,农业农村部动物疾病临床诊疗技术重点实验室/国家级动物医学实验教学中心,内蒙古呼和浩特010010 [2]内蒙古阿拉善盟额济纳旗动物疫病预防控制中心,内蒙古额济纳旗735400 [3]济南市长清区畜牧兽医事业发展中心,山东济南250399

出　　处：《中国兽医科学》2023年第9期1158-1165,共8页Chinese Veterinary Science

基　　金：国家自然科学基金资助项目(31960724,32160838,31460656);内蒙古自治区重点研发和成果转化计划项目(2023YFDZ0048);内蒙古自然科学基金资助项目(2021MS03088,2019MS03017,2023LHMS03005,2023LHMS03022);内蒙古自治区高等学校科学技术研究项目(NJZY21495)。

摘　　要：为研究卵寄生性真菌——厚垣普可尼亚菌(P.chlamydosporia)发酵液中胞外酶的活性及其杀虫作用,使用不同营养成分组成的液体培养基及诱导物对P.chlamydosporia进行培养和诱导,测定P.chlamydosporia发酵液中蛋白质含量、胞外水解酶活性及发酵液对秀丽新杆线虫及马副蛔虫卵的毒性作用,确定产生胞外蛋白质和具杀虫作用物质的最适液体培养基与诱导物种类。结果显示,马副蛔虫卵诱导的枸橼酸培养基发酵液中蛋白质种类最丰富且质量浓度最高,可达3.37±0.03 g/L,蛋白酶活性(2.00±0.06 U/m L)与磷酸酶活性[酸性磷酸酶(28.58±1.30 U/L)、碱性磷酸酶(12.68±1.57 U/L)]均较强,对秀丽新杆线虫的致死率为73.91%±1.72%,同时对马副蛔虫卵的孵化也具有一定抑制作用,相对孵化抑制率可达64.06%±2.46%,均显著高于对照组。结果表明,不同培养基及诱导物对P.chlamydosporia的代谢过程产生显著影响,导致其分泌的蛋白质种类不同,其质量浓度和酶活性均有显著区别(P<0.05)。发酵液不仅具有杀线虫活性,还有良好的抑制虫卵孵化作用。上述结果为进一步研究卵寄生性真菌的杀虫作用与机理提供了重要参考依据。To study the production of extracellular enzyme and their nematode killing effects of the fermentation broth of the egg destroying fungus-Pochonia chlamydosporia,liquid media composed of different nutrients and inducers were used to culture P.chlamydosporia,followed by measuring the protein content,extracellular hydrolase activity in P.chlamydosporia fermentation broth and the toxic effects of P.chlamydosporia fermentation broth on Caenorhabditis elegans and the toxicity of Parascaris equorum eggs,then the optimal liquid medium and inducers to produce extracellular proteins and nematode killing effects were determined.The results showed that,there were the most abundant and the highest mass concentration of proteins,which was 3.37±0.03 g/L,while the protease activity was 2.00±0.06 U/mL.Besides,the phosphatase activity was also the highest in the fermentation induced by Parascaris equorum eggs,with acidic phosphatase activity was 28.58±1.30 U/L and alkaline phosphatase activity was 12.68±1.57 U/L.The lethality rate of the fermentation forC.elegans was 73.91%±1.72%,meanwhile the fermentation liquid also had inhibitory effect on the hatching of the Parascaris equorum eggs,with the relative inhibition rate up to 64.06%±2.46%,which were significantly higher than the corresponding control group.The results showed that different media and inducers had a significant effect on the metabolic process of P.chlamydosporia,resulting in different protein secreted,and significantly different in protein concentration and enzyme activity(P<0.05).There was not only nematode activity but also good inhibition of egg hatching.The above results would provide an important reference for further studying on the nematode killing mechanism of the fungus.

关 键 词：卵寄生性真菌 厚垣普可尼亚菌 胞外蛋白质 胞外水解酶活性 杀虫作用 

分 类 号：S852.66[农业科学—基础兽医学]