哈尔滨两种斯氏线虫及其共生菌的分离鉴定和低温侵染力分析  

Isolation and Identification of Entomopathogenic Nematodes and Their Symbiotic Bacteria from Harbin(Northeast China) and Infectivity of Entomopathogenic Nematodes in Low Temperature

作　　者：吕洋[1] 韩岚岚[1] 赵奎军[1] 宋捷[1] 朱明贺 

机构地区：[1]东北农业大学农学院,哈尔滨150030

出　　处：《中国生物防治学报》2014年第4期558-563,共6页Chinese Journal of Biological Control

基　　金：现代农业产业技术体系建设专项(CARS-04);公益性行业(农业)科研专项(201103002)

摘　　要：从哈尔滨当地土壤中分离出2个斯氏线虫品系。并通过分子生物学技术对这2种斯氏线虫的内转录间隔区(ITS)进行PCR扩增和序列分析,明确这2种哈尔滨斯氏线虫分别是海滨斯氏线虫Steinernema litorale和拟双角斯氏线虫Steinernema ceratophorum,其扩增的DNA片段长度分别为913和1000 bp。关于海滨斯氏线虫的共生菌研究目前尚未见报道,进而对其共生菌进行了分离鉴定,通过扩增核糖体16S rDNA片段,确定海滨斯氏线虫携带的共生菌为伯氏杆菌Xenorhabdus bovienii,其扩增片段大小为1388 bp;拟双角斯氏线虫携带的共生菌为Xenorhabdus budapestensis,其扩增片段大小为1517 bp。为了验证海滨斯氏线虫及拟双角斯氏线虫在高寒地区的活力,在低温条件下对大蜡螟的侵染力进行了测定。结果表明,5℃条件下侵染144 h,4种线虫处理的大蜡螟幼虫均未死亡;10℃条件下,海滨斯氏线虫表现出明显较高的侵染力,在处理96 h大蜡螟幼虫的校正死亡率为85.83%,其他3个处理校正死亡率都为0;15℃条件下,拟双角斯氏线虫表现出明显较高的侵染力,在处理72 h大蜡螟的校正死亡率为89.17%,海滨斯氏线虫在处理72 h大蜡螟未出现死亡,处理96 h开始出现死亡,且校正死亡率为87.5%,而拟双角斯氏线虫在处理96 h后校正死亡率为96.67%,拟双角斯氏线虫表现出较高的侵染力显著高于其他2种处理,但与海滨斯氏线虫相比,二者无显著差异。Steinernema litoraleandSteinernemaceratophorumwere isolated from a soil sample collected from Harbin of Heilongjiang province and identified by molecular approach. The results of PCR amplification and sequence analysis of these twoSteinernema internal transcribed spacer （ITS）, indicated that the fragment length of ITS region ofS. litoraleandS.ceratophorum were 913 and 1000 bp, respectively. There is no report about symbiotic bacteria ofS. litorale research. So we isolated and identified its symbiotic bacteria through amplifying 16S rDNA fragment, the symbiotic bacteria ofS. litorale wasXenorhabdus bovienii with 1388 bp and the symbiotic bacteria of S. ceratophorum wasXenorhabdus budapestensis with 1517 bp. In order to clarify S. litorale and S. ceratophorumvitality in the alpine region, we analyzed the infectivity of the both symbiotic bacteria to infest Galleria mellonella. The results showed that theG. mellonella larvaes no died when they were infested the four kinds nematodes at 5℃ condition for 144 h.G. mellonella infected byS. litorale was significantly higher molitalythan other EPNs at 10℃, the corrective mortality ofG. mellonellalarvae being treated for 96 h was 85.83%, and the other three treated corrective mortalities were 0 （P〈0.05）;at 15℃,S. ceratophorum appeared an relatively high infectivity, the corrective mortality ofG. mellonella larvae being treated for 72 h was 89.17%, andS. litorale did not die after treating G.mellonella larvae for 72 h, and started to die after treatingG. mellonella larvae for 96 h, and its corrective mortality was 87.5%, the corrective mortality ofS. ceratophorumafter treatingG.mellonella larvae for 96 h was 96.67%,S. ceratophorum appeared relatively high infectivity than the other two treatments, however, while comparing toS. litorale, there was no significant difference between them.

关 键 词：斯氏线虫 分离鉴定 低温 侵染力 

分 类 号：S476[农业科学—农业昆虫与害虫防治]