摘要
【目的】筛选球孢白僵菌并探究其对长林小蠹幼虫和成虫的致病性,以及分析球孢白僵菌对长林小蠹幼虫和成虫的侵染过程。【方法】1)通过形态学鉴定和分子生物学鉴定确定所筛选菌株的品系;2)利用Excel2016软件计算累计校正死亡率,SPSS 26.0软件进行方差分析,并计算毒力回归方程式和LC50;3)利用扫描电镜观察球孢白僵菌侵染长林小蠹幼虫和成虫的过程。【结果】1)经鉴定,从长林小蠹僵虫虫体上分离纯化得到的是4株不同的球孢白僵菌(编号分别为Bbz1、Bbz2、Bbz3和Bbz4);2)对长林小蠹幼虫和成虫进行毒力测定,4株球孢白僵菌对长林小蠹幼虫和成虫毒力并不相同,其中,菌株Bbz4对幼虫的毒杀效果最好,接种7 d后,1×10^(8)孢子/m L浓度的孢子悬浮液对长林小蠹幼虫致死率达到100%,LC50为3.68×10^(5)孢子/m L,对成虫毒杀效果最好的是菌株Bbz1,接种7 d后,1×10^(8)孢子/m L浓度的孢子悬浮液对长林小蠹成虫致死率达到100%,LC_(50)为3.27×10^(6)孢子/m L;3)使用菌株Bbz4和菌株Bbz1对长林小蠹幼虫和成虫的侵染进行超微结构观察发现,在接种0~12 h时,菌株分生孢子接触并附着在长林小蠹体表;接种18 h时,发现孢子萌发并产生芽管;接种36 h时,发现有菌丝向内穿透体壁的现象出现;接种72 h时,菌丝伸出体外;接种96~120 h时,整个虫体被菌丝覆盖。【结论】确定了从长林小蠹僵虫虫体上分离到的是4株不同球孢白僵菌,其中菌株Bbz4和Bbz1分别对长林小蠹幼虫和成虫致病性最强,并确定接种后不同时间节点上白僵菌的侵染进程。
【Objective】Beauveria bassiana was screened and its pathogenicity on larvae and adults of Hylurgus ligniperda was investigated,the infection process of B.bassiana on larvae and adults of H.ligniperda was verified.【Method】1)Morphological identification and molecular biology identification were used to determine the strain of the selected strains;2)The cumulative corrected mortality rate was calculated using Excel 2016 and the SPSS 26.0 was used for ANOVA and the virulence regression equation and LC50 were calculated;3)Electron microscopy scanning was used to observe the infection of larvae and adults of H.ligniperda.【Result】1)Through bioassay,it was found that 4 strains of B.bassiana were not the same virulence to the larvae and adults of H.ligniperda,named Bbz1,Bbz2,Bbz3 and Bbz4;2)Among them,the strain Bbz4 had the best poisoning effect on larvae,after inoculation for 5 days,the killing effect of 1×10^(8) spores/mL concentration of spore suspension on the larvae of H.ligniperda reached 100%,LC50 was 3.68×10^(5) spores/mL.The best poisoning effect on the adult of H.ligniperda was the strain Bbz1.After inoculation for 5 days,the killing effect of 1×10^(8) spores/mL concentration of spore suspension on the alduts of H.ligniperda reached 100%,and the LC50 was 3.27×10^(6) spores/mL;3)Scanning electron microscopy was performed on the infection of larvae and adults of H.ligniperda used strain Bbz4 and strain Bbz1,and it was found that the conidia of these strains touched and attached to the body surface of H.ligniperda during 0-12 h after inoculation.When inoculated for 18 h,spore germination was found and bud tubes were produced;At 36 h,there was mycelium penetrating the body wall;When inoculated for 72 h,hyphae appeared on the surface of the worm;At 96-120 h of inoculation,the entire body was covered with hyphae.【Conclusion】It was determined that four different strains of B.bassiana were isolated from the body of H.ligniperda,among which strains Bbz4 and Bbz1 were the most pathogenic to the larvae
作者
张瀚文
陈汝婷
许建娇
周俊华
石鹏
杨青山
韩俊
初奎
宇佳
迟德富
ZHANG Hanwen;CHEN Ruting;XU Jianjiao;ZHOU Junhua;SHI Peng;YANG Qingshan;HAN Jun;CHU Kui;YU Jia;CHI Defu(Key Laboratory of Sustainable Management of Forest Ecosystems of the Ministry of Education,Northeast Forestry University,Harbin 150000,Heilongjiang,China;Forestry Station of Dayao Street,Muping District,Yantai City,Yantai 264100,Shandong,China;Forest Resources Monitoring and Protection Service Center of Muping District,Yantai City,Yantai 264100,Shandong,China)
出处
《中南林业科技大学学报》
CAS
CSCD
北大核心
2024年第2期62-72,共11页
Journal of Central South University of Forestry & Technology
基金
国家重点研发计划项目(2021YDF1400300)。
关键词
球孢白僵菌
长林小蠹
扫描电镜
侵染过程
致病性
Beauveria bassiana
Hylurgus ligniperda
scanning electron microscope
infestation process
pathogenicity
