[1]马琪琪,胡宗伟,夏鹏亮,等.防治茄二十八星瓢虫Bt菌株的筛选及杀虫活性分析[J].北方园艺,2021,(14):31-38.[doi:10.11937/bfyy.20204881]
　MA Qiqi,HU Zongwei,XIA Pengliang,et al.Screening of Bacillus thuringiensis Strains and Analysis of Their Insecticidal Activity Against Henosepilachna vigintioctopunctata[J].Northern Horticulture,2021,(14):31-38.[doi:10.11937/bfyy.20204881]
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防治茄二十八星瓢虫Bt菌株的筛选及杀虫活性分析

《北方园艺》[ISSN:1001-0009/CN:23-1247/S] 卷: 期数: 2021年14 页码: 31-38 栏目: 出版日期: 2021-07-30
Title:
Screening of Bacillus thuringiensis Strains and Analysis of Their Insecticidal Activity Against Henosepilachna vigintioctopunctata
作者:
马琪琪1; 胡宗伟1; 夏鹏亮2; 冯万祖1; Natasha I T MBIZA1; 张建民1
（1.长江大学 农学院，湖北 荆州 434025；2.湖北省恩施州烟草公司，湖北 恩施 445000）
Author(s):
MA Qiqi1; HU Zongwei1; XIA Pengliang2; FENG Wanzu1; NATASHA I T Mbiza1; ZHANG Jianmin1
(1.College of Agriculture,Yangtze University,Jingzhou,Hubei 434025;2.Enshi Tobacco Company,Enshi,Hubei 445000)
关键词:
苏云金芽孢杆菌; 茄二十八星瓢虫; 菌株筛选; cry基因; 杀虫活性
Keywords:
Bacillus thuringiensis; Henosepilachna vigintioctopunctata; strains screening;  cry gene; insecticidal activity
DOI:
10.11937/bfyy.20204881
文献标志码:
A
摘要:
以从湖北宜昌、襄阳、荆州、广东东莞、安徽安庆5个地区采集得到的52份土样为试材，分别采用温度筛选法、镜检法、PCR-RFLP法和SDS-PAGE等方法，分离了土壤中的苏云金芽孢杆菌，并鉴定了分离出的Bt菌株的基因型，通过茄二十八星瓢虫二龄幼虫的室内杀虫试验测定分离株的杀虫活性，同时分析了部分Bt菌株的伴孢晶体蛋白，以期挖掘新的Bt菌株资源，筛选出对茄二十八星瓢虫具有高毒力的Bt菌株。结果表明：分离得到10株含cry3基因的菌株，3株含cry7基因的菌株和2株含cry8基因的菌株，得到1株对茄二十八星瓢虫二龄幼虫有高毒力的Bt菌株，当其伴孢晶体悬浮液的浓度在40 mg?mL-1时，校正死亡率最高可达40.42%；发现YC-2-20、GD-4-1均表达分子量约为67 kDa的晶体蛋白；GD-5-7表达分子量约为130 kDa和80 kDa的晶体蛋白；YC-12-6表达分子量约为130 kDa和70 kDa的晶体蛋白。试验筛选出的Bt菌株为农业生产中茄二十八星瓢虫的绿色防控提供了可应用的菌株及基因资源，同时对延缓害虫抗药性的产生也具有重要意义。
Abstract:
Taking 52 soil samples as test materials,soil samples were collected from five areas:Yichang,Xiangyang,Jingzhou in Hubei Province;Dongguan in Guangdong Province;Anqing in Anhui Province.The temperature screening method,microscopic examination method,PCR-RFLP and SDS-PAGE were used as experimental methods.The Bt in the soil was isolated,and the genotype of the isolated Bt strain was identified.The insecticidal activity of the isolated strain was determined by indoor insecticidal test for the second instar larvae of Henosepilachna vigintioctopunctata,and the crystal protein of some Bt strains was analyzed.In order to discover new Bacillus thuringiensis strains resources.The results showed that a total of ten strains containing cry3 gene,three strains containing cry7 gene and two strains containing cry8 gene were isolated,and one strain of Bt strain with high virulent to the second instar larva of Henosepilachna vigintioctopunctata was obtained.When the concentration of its adjoint crystal suspension was 40 mg?mL-1,the corrected mortality rate could reach 40.42%.It was found that YC-2-20 and GD-4-1 both expressed crystal proteins with molecular weight of about 67 kDa.GD-5-7 expressed crystal proteins with molecular weights of about 130 kDa and 80 kDa.YC-12-6 expresses crystal proteins with molecular weights of about 130 kDa and 70 kDa.The Bt strains screened out in the experiment provided applicable strains and genetic resources for the green prevention and control of Henosepilachna vigintioctopunctata in agricultural production,and also had important significance for delaying the emergence of pest resistance.

参考文献/References:

[1]杨会玲,韩魁魁,赵唯,等.茄二十八星瓢虫的发生与综合防治[J].西北园艺(蔬菜),2011(6):36-37.[2]涂小云,王国红.茄二十八星瓢虫生物防治研究进展[J].中国植保导刊,2010,30(3):13-16.[3]〖JP2〗刘瑶.茄子病虫害综合防治措施[J].吉林蔬菜,2019(2):43.〖JP〗[4]MARTINEZ C,CABALLERO P.Contents of cry genes and insecticidal toxicity of Bacillus thuringiensis strains from terrestrial and aquatic habitats[J].Journal of Applied Microbiology,2002,92(4):745-752.[5]HERNANDEZ C,ANDREW R,BEL Y,et al.Isolation and toxicity of Bacillus thuringiensis from potato-growing areas in Bolivia[J].Journal of Invertebrate Pathology,2005,88(1):8-16.[6]罗兰.苏去金杆菌的分离、cry/cyt基因鉴定及杀线虫活性的研究[D].杨凌:西北农林科技大学,2005.[7]HUANG D F,ZHANG J,SONG F P,et al.Microbial control and biotechnology research on Bacillus thuringiensis in China[J].Journal of Invertebrate Pathology,2007,95(3):175-180.[8]杨佳,姚瑶.Bt基因及其在抗鞘翅目害虫上的应用[J].吉林农业,2019(10):67.[9]张杰,宋福平,李长友,等.对鞘翅目害虫高毒力Bt基因cry3Aa7的分离克隆及表达研究[J].中国农业科学,2002(6):650-653.[10]刘艳菊.发根农杆菌介导的Bt基因Cry8e转化大豆抗蛴螬研究[D].新乡:河南师范大学,2010.[11]方梅,谢建坤,朱敏,等.光周期和LED光源对茄二十八星瓢虫成虫行为的影响[J].昆虫学报,2018,61(11):1295-1299.[12]CRAWFORD I T,GREIS K D,PARKS L,et al.Facile autoplast generation and transformation in Bacillus thuringiensis subsp.kurstaki[J].Journal of Bacteriology,1987,169(12):5423-5428.[13]崔莹莹,席宇,杨艳坤,等.一株对烟草甲高毒力的苏云金芽孢杆菌的分离与鉴定[J].化学与生物工程,2009,26(5):42-45.[14]李云龙.对鞘翅目昆虫高毒力苏云金芽孢杆菌的筛选鉴定[D].哈尔滨:黑龙江大学,2012.[15]谢柳,张文飞,全嘉新,等.广西大王岭和大明山自然保护区苏云金芽孢杆菌收集与鉴定[J].基因组学与应用生物学,2009,28(1):62-68.[16]张路路.苏云金芽孢杆菌的分离、筛选、鉴定及发酵条件优化[D].海口:海南大学,2014.[17]初立良,郑桂玲,周洪旭,等.两株杀鞘翅目害虫Bt菌株的生物活性及杀虫蛋白基因鉴定[J].华北农学报,2010,25(3):235-238.[18]李帅,张茜,高剑,等.牡丹江火山口原始森林苏云金芽孢杆菌的分离和cry基因鉴定[J].牡丹江师范学院学报(自然科学版),2016(4):47-49.[19]谷有芳.西藏土壤苏云金芽孢杆菌菌株分离及基因型鉴定研究[D].哈尔滨:黑龙江大学,2014.[20]MEIHIAR M,AHMAD M,AL-ZYOUD F,et al.Environmental distribution,frequency and toxicity of Bacillus thuringiensis in syria[J].Annual Research & Review in Biology,2015,40(40):141-158.[21]谢本贵,王香萍,李传仁,等.茄二十八星瓢虫耐饥能力及饥饿对雌成虫寿命和繁殖的影响[J].世界生态学,2017,6(1):5.

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备注/Memo

第一作者简介：马琪琪（1995-），女，硕士研究生，研究方向为植物抗虫基因工程。E-mail：1846047269@qq.com.责任作者：张建民（1974-），男，博士，教授，硕士生导师，现主要从事植物抗虫基因工程等研究工作。E-mail：371995966@qq.com.基金项目：国家自然科学基金资助项目（31471783）。收稿日期：2020-11-19

更新日期/Last Update: 2021-10-27