两株石榴内生细菌对松材线虫的生防效果和鉴定  

The Biocontrol Efficacy of two Strains of Endophytic Bacteria from Pomegranate on Bursaphelenchus xylophilus and their Identification

作　　者：雷秋艳 王国丽 刘冬梅 贺小如 于金英 汪冶[1] LEI Qiuyan;WANG Guoli;LIU Dongmei;HE Xiaoru;YU Jinying;WANG Ye(School of Ecology,Lishui University,Lishui 323000,Zhejiang;College of Life Science and Medicine,Zhejiang Sci-Tech University,Hangzhou 310018,Zhejiang)

机构地区：[1]丽水学院生态学院,浙江丽水323000 [2]浙江理工大学生命科学与医药学院,浙江杭州310018

出　　处：《丽水学院学报》2024年第5期31-40,共10页Journal of Lishui University

基　　金：丽水市莲都区森林病虫防治检疫站委托项目“微生物源生物农药防治松材线虫病研究”(202207SF15)。

摘　　要：为寻找对松材线虫具毒杀活性的植物内生细菌,对石榴进行内生细菌分离纯化,共得到13株内生细菌,采用浸渍法测定细菌发酵滤液的杀线活性,并通过松枝水培法对活性菌株发酵滤液抗松材线虫病进行评价。结果显示,松材线虫经菌株YL07和YL08的发酵滤液处理24 h后,校正死亡率均达100%,松材线虫体内过氧化氢酶(CAT)、乙酰胆碱酯酶(AChE)、谷胱甘肽转移酶(GST)的活性显著低于对照组。水培实验中,菌株YL07、YL08的发酵滤液处理组和CK1(未接松材线虫+蒸馏水)处理组处理12 d后,未出现任何病理症状(病情指数为0),而CK3(接松材线虫+LB培养基)和CK2(接松材线虫+蒸馏水)处理组出现萎蔫病理症状(病情指数分别为0.417和0.917)。通过形态学特征、生理生化特征、16S r DNA序列及其系统发育分析,鉴定菌株YL07和YL08分别为蜡样芽孢杆菌(Bacillus cereus)和暹罗芽孢杆菌(Bacillus siamensis)。综上所述,菌株YL07和YL08对松材线虫具显著毒杀作用,且在发酵滤液的处理下松枝表现出抗病性,对松材线虫病有一定的防治效果。To searched for endophytic bacteria in plants that have toxic activity against pine wood nematodes,pomegranate was selected for the isolation and purification of endophytic bacteria,and a total of 13 strains of endophytic bacteria were obtained.The bacterium fermentation filtrate was tested for its nematocidal activity with the immersion method.The anti-pine wilt disease effect of the fermentation filtrate of the active strain was evaluated by using the pine branch hydroponic method.The results showed that the corrected mortality rate of Bursaphelenchus xylophilus treated with fermentation filtrate of strains YL07 and YL08 for 24 hours reached 100%.The activities of catalase(CAT),acetylcholinesterase(AChE),and glutathione transferase(GST)in B.xylophilus were significantly lower than those in the control group.In the hydroponic experiment,after 12 days of treatment with the fermentation filtrate of strains YL07 and YL08 and the CK1(no inoculation with pine wood nematode+distilled water)treatment group,no pathological symptoms(disease index of 0)were observed,while the CK3(inoculation with pine wood nematode+LB medium)and CK2(inoculation with pine wood nematode+distilled water)treatment groups showed wilting pathological symptoms(disease index of 0.417 and 0.917,respectively).Through morphological,physiological and biochemical characteristics,16S rDNA sequence and phylogenetic analysis,strains YL07 and YL08 were identified as Bacillus cereus and Bacillus siamensis,respectively.In summary,strains YL07 and YL08 have significant toxic effects on pine wood nematodes,and under the treatment of fermentation filtrate,pine branches exhibit disease resistance,which has certain control effects on pine wilt disease.

关 键 词：内生细菌 松材线虫 杀线活性 

分 类 号：S763.3[农业科学—森林保护学]