对草地贪夜蛾高毒力的苏云金杆菌菌株筛选

doi:10.16409/j.cnki.2095-039x.2019.05.028

中国生物防治学报 ›› 2019, Vol. 35 ›› Issue (5): 721-728.DOI: 10.16409/j.cnki.2095-039x.2019.05.028

• 草地贪夜蛾生物防治专栏 • 上一篇下一篇

对草地贪夜蛾高毒力的苏云金杆菌菌株筛选

刘华梅¹, 胡虓¹, 王应龙², 杨普云³, 束长龙², 朱晓明³, 张杰², 孙刚忠¹, 张晓明¹, 李青¹

1. 武汉科诺生物科技股份有限公司, 武汉 430074;
2. 中国农业科学院植物保护研究所/植物病虫害生物学国家重点实验室, 北京 100193;
3. 全国农业技术推广服务中心, 北京 100125

收稿日期:2019-08-21 出版日期:2019-10-08 发布日期:2019-10-26
通讯作者: 李青,硕士,副研究员,E-mail:liqing@biokn.com。
作者简介:刘华梅,博士,副高级工程师,E-mail:liuhuamei@biokn.com
基金资助:
国家重点研发计划（2017YFD0201204）

Screening for Bacillus thuringiensis Strains with High Toxicity against Spodoptera frugiperda

LIU Huamei¹, HU Xiao¹, WANG Yinglong², YANG Puyun³, SHU Changlong², ZHU Xiaoming³, ZHANG Jie², SUN Gangzhong¹, ZHANG Xiaoming¹, LI Qing¹

1. Wuhan Kernel Bio-tech Co., Ltd, Wuhan 430074, China;
2. State Key Laboratory for Biology of Plant Diseases and Insect Pests/Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China;
3. National Agro-Tech Extension and Service Center, Beijing 100125, China

Received:2019-08-21 Online:2019-10-08 Published:2019-10-26

摘要/Abstract

摘要： 筛选对草地贪夜蛾高毒力Bt菌株，对于开发绿色高效防治草地贪夜蛾新产品具有重要意义。本研究从自行筛选的对夜蛾科害虫高毒力的Bt菌株资源中，选取9株菌株进行草地贪夜蛾室内毒力测定，并对高毒力菌株进行杀虫基因分析；选取室内毒力测定活性最高的3株菌株进行田间小区药效试验。结果表明，9株Bt菌株在药后7 d对草地贪夜蛾初孵幼虫均表现出显著的毒杀效果，LC₅₀范围在0.07～3.38 μg/g，其中毒力最高的3个菌株分别是KN50、KN11和KNR8，LC₅₀分别为0.07、0.23和0.43 μg/g，表现非常显著的致死效果；对KN50、KN11和KNR8等6种高毒力的Bt菌株的杀虫基因分析发现，他们都含有vip3Aa、cry2Ab、cry1Ac、cry1Ia杀虫基因。KN50、KN11和KNR8菌株的32000 IU/mg制剂在田间对草地贪夜蛾表现出不同程度防效，其防效高低与室内毒力结果一致，其中KN50药后7 d防效最高，在200和400 g/667 m²下对1～3龄幼虫防效分别可达86.09%和95.22%，对不同龄期幼虫混发的防效为72.61%和86.59%。因此KN50、KN11和KNR8这3个菌株能作为备选高效菌株用于防治草地贪夜蛾新产品开发，为实现草地贪夜蛾的绿色防控、化学农药合理减替提供重要保障。

关键词: 草地贪夜蛾, 苏云金杆菌, 毒力测定, 杀虫基因, 田间防效

Abstract: To screen Bt strains with high virulence against Spodoptera frugiperda is crucial for the development of green and efficient IPM of S. frugiperda. In this study, nine Bt strains with high toxicity to Noctuidae, isolated by our company, were tested for their virulence against S. frugiperda in laboratory, and analyzed the insecticidal genes. Three of the highest virulent strains were further tested in field trial for their control efficacies against S. frugiperda. The results indicated that nine Bt strains showed prominent insecticidal activity against neonate of S. frugiperda at 7 days post treatment. The LC₅₀ ranged from 0.07 to 3.38 μg/g. The three highest virulent strains were KN50, KN11 and KNR8, with LC₅₀ of 0.07, 0.23 and 0.43 μg/g, respectively, showing very remarkable efficacy. Insecticidal gene analysis results demonstrated that six highly virulent Bt strains contained the same four insecticidal genes-vip3Aa, cry2Ab, cry1Ac and cry1Ia. Field trial results showed that the control efficacies of KN50, KN11 and KNR8 with 32000 IU/mg were significantly different, but were consistent with laboratory results. KN50 showed the highest insecticidal efficacy at 7 days post treatment, which was 86.09% and 95.22% for 200 g/667 m² and 400 g/667 m², respectively, against the first to third instar larvae, and 72.61% and 86.59% against larval populations of mixed instars. Therefore, the KN50, KN11 and KNR8 strains represent candidates for new product development to replace chemical pesticides and achieve green control S. frugiperdar.

Key words: Spodoptera frugiperda, Bacillus thuringiensis, bioassay, insecticidal gene analysis, field control efficacy

中图分类号:

S476.1

刘华梅, 胡虓, 王应龙, 杨普云, 束长龙, 朱晓明, 张杰, 孙刚忠, 张晓明, 李青. 对草地贪夜蛾高毒力的苏云金杆菌菌株筛选[J]. 中国生物防治学报, 2019, 35(5): 721-728.

LIU Huamei, HU Xiao, WANG Yinglong, YANG Puyun, SHU Changlong, ZHU Xiaoming, ZHANG Jie, SUN Gangzhong, ZHANG Xiaoming, LI Qing. Screening for Bacillus thuringiensis Strains with High Toxicity against Spodoptera frugiperda[J]. Chinese Journal Of Biological Control, 2019, 35(5): 721-728.

参考文献

[1] Cruz I, Figueiredo M C, Silva R B, et al. Using sex pheromone traps in the decision-making process for pesticide application against fall armyworm, (Spodoptera frugiperda (Smith) (Lepidoptera:Noctuidae) larvae in maize[J]. International Journal of Pest Management, 2012, 58(1):83-90.
[2] Nagoshi R N, Rosas-Garcia N M, Meagher R L, et al. Haplotype profile comparisons between Spodoptera frugiperda (Lepidoptera:Noctuidae) populations from Mexico with those from Puerto Rico, South America, and the United States and their implications to migratory behavior[J]. Journal of Economic Entomology, 2015, 108(1):135-144.
[3] 杨学礼, 刘永昌, 罗茗钟, 等. 云南省江城县首次发现迁入我国西南地区的草地贪夜蛾[J]. 云南农业, 2019(1):72.
[4] 于文静. 我国夏粮增产成定局秋粮丰收有希望[EB/OL].[2019-06-26]. http://www.xinhuanet.com/2019-06/26/c_1124674399.htm
[5] 杨普云, 朱晓明, 郭井菲, 等. 我国草地贪夜蛾的防控对策与建议[J]. 植物保护, 2019, 45(4):1-6.
[6] 姜玉英, 刘杰, 朱晓明. 草地贪夜蛾侵入我国的发生动态和未来趋势分析[J]. 中国植保导刊, 2019, 39(2):33-35.
[7] Viteri D M, Linares A M, Cabrera I, et al. Presence of corn earworm and fall armyworm (Lepidoptera:Noctuidae) populations in sweet corn and their susceptibility to insecticides in Puerto Rico[J]. Florida Entomologist, 2019, 102(2):451-454.
[8] Carvalho R A, Omoto C, Field L M, et al. Investigatingthe molecular mechanisms of organophosphate and pyrethroid resistance in the fall armyworm Spodopterafrugiperda[J]. PLoS ONE, 2013, 8:1-11.
[9] Monnerat R, Martins E, Macedo C, et al. Evidence of field-evolved resistance of Spodoptera frugiperda to Bt corn expressing Cry1F in Brazil that is still sensitive to modified Bt toxins[J]. PLoS ONE, 2015, 10:1-12.
[10] Zhu Y C, Blanco C A, Portilla M, et al. Evidence of multiple/cross resistance to Bt and organophosphate insecticides in Puerto Rico population of the fall armyworm, Spodoptera frugiperda[J]. Pesticide Biochemistry and Physiology, 2015, 122:15-21.
[11] Santos-Amaya O F, Tavares C S, Monteiro H M, et al. Genetics basis of Cry1F resistance in two Brazilian populations of fall armyworm[J]. Crop Protection, 2016, 81(3):154-162.
[12] 王磊, 陈科伟, 钟国华, 等. 重大入侵害虫草地贪夜蛾发生危害、防控研究进展及防控策略探讨[J]. 环境昆虫学报, 2019(3):479-487.
[13] Sanchis V. From microbial sprays to insect-resistant transgenic plants:History of the biospesticide Bacillus thuringiensis. A review[J]. Agronomy for Sustainable Development, 2011, 31(1):217-231.
[14] 束长龙, 张风娇, 黄颖, 等. Bt杀虫基因研究现状与趋势[J]. 中国科学(生命科学), 2016, 46(5):548-555.
[15] 徐国升, 王翠翠, 张贤, 等. 两种不同化学农药抗性背景小菜蛾对BtCry1Ac蛋白敏感度的比较[J]. 中国生物防治学报, 2017, 33(6):726-731.
[16] Raymond B, Johnston P R, Nierlsen-Leroux C, et al. Bacillus thuringiensis:an impotent pathogen?[J]. Trends in Microbiology, 2010, 18(5):189-194.
[17] Andrade R, Rodriguez C, Oehlschlager A C. Optimization of a pheromone lure for Spodopterafrugiperda (Smith) in Central America[J]. Journal of the Brazilian Chemical Society, 2001, 11(6):609-613.
[18] Sanahuja G, Banakar R, Twyman R M. Bacillus thuringiensis:a century of research, development and commercial applications[J]. Plant Biotechnology Journal, 2011, 9:283-300.
[19] 中华人民共和国国家质量监督检验检疫总局, 中国国家标准化管理委员会. GB/T 19567.3-2004苏云金芽胞杆菌可湿性粉剂[S]. 北京:中国标准出版社, 2004.
[20] 唐启义, 冯明光. DPS数据处理系统[M]. 北京:科学出版社, 2006.
[21] Li Y Q, Shu C L, Shan Y M, et al. Complete genome sequence of Bacillus thuringiensis Bt185, a potential soil insect biocontrol agent[J]. Journal of Integrative Agriculture, 2017, 16(3):749-751.
[22] 农业农村部.农业农村部办公厅关于做好草地贪夜蛾应急防治用药有关工作的通知[EB/OL].[2019-06-3]. http://www.moa.gov.cn/gk/tzgg_1/tfw/201906/t20190605_6316201.htm.
[23] Monnerat R G, Batista A C, TellesdeMedeiros P, et al. Screening of Brazilian Bacillus thuringiensis isolates active against Spodoptera frugiperda, Plutella xylostella and Anticarsia gemmatalis[J]. Biological Control, 2007, 41(3):291-295.
[24] Santos K B, Neves P, MeneguimA M, et al. Selection and characterization of the Bacillus thuringiensis strains toxicto Spodoptera eridania (Cramer), Spodoptera cosmioides (Walker) and Spodoptera frugiperda (Smith) (Lepidoptera:Noctuidae)[J]. Biological Control, 2009, 50(2):157-163.
[25] Escudero, I R, Banyuls N, Bel Y, et al. A screening of five Bacillus thuringiensis Vip3A proteins for their activity against lepidopteran pests[J]. Journal of Invertebrate Pathology, 2014, 117(3):51-55.
[26] Fatoretto J C, Michel A P, Silva Filho M C, et al. Adaptive potential of fall armyworm (Lepidoptera:Noctuidae) limits Bttrait durability in Brazil[J]. Journal of Integrated Pest Management, 2017, 8(1):1-10,17.
[27] 张丹丹, 吴孔明. 国产Bt-Cry1Ab和Bt-(Cry1Ab+Vip3Aa)玉米对草地贪夜蛾的抗性测定[J]. 植物保护, 2019, 45(4):54-60.
[28] 李国平, 姬婷婕, 孙小旭, 等. 入侵云南草地贪夜蛾种群对5种常用Bt蛋白的敏感性评价[J]. 植物保护, 2019, 45(3):15-20.

对草地贪夜蛾高毒力的苏云金杆菌菌株筛选

Screening for Bacillus thuringiensis Strains with High Toxicity against Spodoptera frugiperda

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

联系我们

[1]	李贝贝, 田野, 杜桂林, 岳方正, 农向群, 刘廷辉, 张泽华, 王广君. 丝氨酸蛋白酶抑制剂Serpin1对绿僵菌侵染飞蝗的影响[J]. 中国生物防治学报, 2020, 36(5): 729-736.
[2]	田俊策, 鲁艳辉, 王国荣, 郑许松, 杨亚军, 徐红星, 方琦, 叶恭银, 臧连生, 吕仲贤. 种赤眼蜂对草地贪夜蛾卵的寄生能力研究[J]. 中国生物防治学报, 2020, 36(4): 485-490.
[3]	路子云, 杨小凡, 马爱红, 冉红凡, 刘文旭, 李建成. 管侧沟茧蜂对不同日龄草地贪夜蛾幼虫的寄生效果[J]. 中国生物防治学报, 2020, 36(4): 491-495.
[4]	杨磊, 李芬, 吴少英. 草地贪夜蛾寄生蜂资源及其调控寄主免疫反应的研究[J]. 中国生物防治学报, 2020, 36(4): 496-506.
[5]	黄潮龙, 汤印, 何康来, 王振营. 双斑青步甲幼虫对草地贪夜蛾幼虫的捕食能力[J]. 中国生物防治学报, 2020, 36(4): 507-512.
[6]	李萍, 李玉艳, 向梅, 王孟卿, 毛建军, 陈红印, 张礼生. 大草蛉幼虫对草地贪夜蛾低龄幼虫的捕食能力评价[J]. 中国生物防治学报, 2020, 36(4): 513-519.
[7]	王燕, 张红梅, 李向永, 尹艳琼, 赵雪晴, 陈福寿, 张礼生. 益蝽不同龄期若虫对草地贪夜蛾幼虫的捕食能力[J]. 中国生物防治学报, 2020, 36(4): 520-524.
[8]	王亚楠, 赵胜园, 何运转, 吴孔明, 李国平, 封洪强. 黄带犀猎蝽对草地贪夜蛾幼虫的捕食作用[J]. 中国生物防治学报, 2020, 36(4): 525-529.
[9]	徐翔, 杨淞杰, 李维强, 衡晓蓉, 尹勇. 绿僵菌油悬浮剂与减量化学农药联用对冬玉米草地贪夜蛾的防控效果[J]. 中国生物防治学报, 2020, 36(4): 530-533.
[10]	蒋春先, 田亚静, 李庆, 王海建, 杨群芳. 杀螨剂对朱砂叶螨的毒力测定和对加州新小绥螨的安全性评价[J]. 中国生物防治学报, 2020, 36(4): 551-557.
[11]	宋健, 曹伟平, 郭庆港, 王猛, 李社增, 杜立新. 苏云金芽胞杆菌JQD117菌株对韭蛆体内几种酶活性的影响[J]. 中国生物防治学报, 2020, 36(4): 558-563.
[12]	苏造堂, 张凌英, 徐天梅, 杜广祖, 陈斌, 肖关丽. 马铃薯块茎蛾幼虫致病黏质沙雷氏菌的分离鉴定及杀虫活性[J]. 中国生物防治学报, 2020, 36(3): 361-370.
[13]	韩光杰, 徐灵环, 李传明, 刘琴, 彭琦, 徐彬, 宋福平, 徐健. 苏云金杆菌Bt-59菌株Cry/Cyt毒素蛋白鉴定及杀虫活性[J]. 中国生物防治学报, 2020, 36(3): 458-464.
[14]	张桂芬, 张毅波, 张杰, 刘万学, 王玉生, 万方浩, 束长龙, 刘慧, 王福莲, 赵林, 李庆红, 王树明, 蒋家强. 苏云金芽胞杆菌G033A对新发南美番茄潜叶蛾的室内毒力及田间防效[J]. 中国生物防治学报, 2020, 36(2): 175-183.
[15]	王红丽, 善文辉, 胡海瑶, 李燕, 王琦, 王坤, 卞凤娥. 生防菌混合接种对葡萄灰霉病菌的防治效果[J]. 中国生物防治学报, 2020, 36(2): 265-271.