椰子织蛾幼虫致病Bt菌的鉴定与杀虫活性测定

doi:10.16409/j.cnki.2095-039x.2016.02.022

中国生物防治学报

• 研究简报 • 上一篇

椰子织蛾幼虫致病Bt菌的鉴定与杀虫活性测定

孙晓东^1,2, 阎伟¹, 李朝绪¹, 吕朝军¹, 覃伟权¹

1. 中国热带农业科学院椰子研究所, 文昌 571339;
2. 东北农业大学生命科学学院, 哈尔滨 150030

收稿日期:2015-06-30 出版日期:2016-04-08 发布日期:2016-04-08
通讯作者: 覃伟权
作者简介:孙晓东(1983-),男,博士,助理研究员,E-mail:sxd1949@163.com
基金资助:
海南省重大项目(ZDZX2013008);海南省重点项目(ZDXM20130049)

Identification of Bacillus thuringiensis from Opisina arenosella and Determination of Its Insecticidal Bioactivity

SUN Xiaodong^1,2, YAN Wei¹, LI Chaoxu¹, LÜ Chaojun¹, TAN Weiquan¹

1. Coconut Research Institute, Chinese Academy of Tropical Agricultural Sciences, Wenchang 571339, China;
2. College of Science, Northeast Agricultural University, Harbin 150030, China

Received:2015-06-30 Online:2016-04-08 Published:2016-04-08

摘要/Abstract

摘要： 从海南省五指山采集的椰子织蛾僵虫上分离得到一株对椰子织蛾幼虫具有毒杀作用的菌株WZS171并对其生物防治潜力进行评测。通过形态学、cry基因的鉴定明确了该菌株为苏云金芽胞杆菌(Bt)。基因鉴定结果表明该菌株含有cry1Aa13、cry1Gb1、cry1D、cry1Ia、cry2Af共5种新型cry基因,NCBI Blast与GenBank中已知基因序列最大相似性分别为98%、99%、90%、92%、99%;生测结果表明72 h后该菌株菌悬液对椰子织蛾幼虫的LC₅₀为9.08 μg/mL,95%置信区间6.72~12.26 μg/mL。该菌株含有5种新型的杀虫编码基因,并且对椰子织蛾幼虫具有较高毒力,生物防治潜力大。

关键词: Bt, 椰子织蛾, 鉴定, 杀虫活性

Abstract: A virulent strain WZS171 with high infectivity was isolated from naturally infected and dead larvae of Opisina arenosella Walker collected from Hainan Wuzhishan. The strain was evaluated for its insecticidal bioactivities. The strain was identified as Bacillus thuringiensis by morphological tests and analysis of cry genes. The new genes of cry1Aa13, cry1Gb1, cry1D, cry1Ia, cry2Af were found in this isolate. NCBI BLAST revealed that the nucleotide fragments of these genes showed homology of 98%, 99%, 90%, 92% and 99%, respectively, with the known genes at the GeneBank. Bioactivity of WZS171 against the larvae of Opisina arenosella Walker was assayed in laboratory. WZS171 showed high insecticidal activity at 72 h post-treatment, with a LC₅₀ of 9.08 μg/mL and a confidence interval of 6.72-12.26 μg/mL at the 95% confidence level. These results show that WZS171 is host specific in virulence and highly virulent against larvae of Opisina arenosella Walker.

Key words: Bacillus thuringiensis, Opisina arenosella Walker, identification of cry gene, insecticidal activity

孙晓东, 阎伟, 李朝绪, 吕朝军, 覃伟权. 椰子织蛾幼虫致病Bt菌的鉴定与杀虫活性测定[J]. 中国生物防治学报, DOI: 10.16409/j.cnki.2095-039x.2016.02.022.

SUN Xiaodong, YAN Wei, LI Chaoxu, LÜ Chaojun, TAN Weiquan. Identification of Bacillus thuringiensis from Opisina arenosella and Determination of Its Insecticidal Bioactivity[J]. journal1, DOI: 10.16409/j.cnki.2095-039x.2016.02.022.

参考文献

[1] Shivashankar T, Annadurai R S, Srinivas M. Control of coconut black-headed caterpillar (Opisina arenosella Walker) by systemic application of ‘Soluneem’-A new water-soluble neem insecticide formulation[J]. Current Science, 2000, 78(2):25.
[2] Venkatesan T, Jalali S K, Srlnivasa M K, et al. A novel method of field release of Goniozus nephantidis (Muesebeck), an important primary parasitoid of Opisina arenosella Walker on coconut[J]. Journal of Biological Control, 2003, 17(1):79-80.
[3] Venkatesan T, Jalali S K, Srinivasa M K. Competitive interactions between Goniozus nephantidis and Bracon brevicornis,parasitoids of the coconut pest Opisina arenosella[J]. International Journal of Pest Management, 2009, 55(3):257-263.
[4] 刘向蕊, 吕宝乾, 金启安, 等. 5种杀虫剂对入侵害虫椰子织蛾的室内毒力测定[J]. 生物安全学报, 2014, 23(1):13-17.
[5] 阎伟, 吕宝乾, 李洪, 等. 椰子织蛾传入中国及其海南省的风险性分析[J]. 生物安全学报, 2013, 22(3):163-168.
[6] 李洪, 阎伟, 刘丽. 新入侵害虫椰子织蛾的发生及防治[J]. 中国森林病虫, 2015, 34(4):10-13.
[7] 吕宝乾, 严珍, 金启安, 等. 警惕椰子织蛾Opisina arenosella Walker (鳞翅目:织蛾科)传入中国[J]. 生物安全学报, 2013, 22(1):17-22.
[8] Aaron J G, Jennifer L P, Eric H C. Field-evolved resistance by western corn rootworm to multiple Bacillus thuringiensis toxins in transgenic maize[J]. PNAS, 2014, 111(14):5141-5146.
[9] 束长龙, 谷少华, 窦黎明, 等. 对小地老虎具有杀虫毒力的苏云金芽胞杆菌菌株的分离及鉴定[J]. 植物保护, 2007, 33(5):41-44.
[10] Eitan B D. Bacillus thuringiensirs subsp. israelensis and its dipteran-specific toxins[J]. Toxins, 2014, 6(4):1222-1243.
[11] 刘楠, 王少丽, 宋福平, 等. Cry1Ba3、Cry1Ia8蛋白对Cry1Ac抗性小菜蛾的杀虫活性研究[J]. 植物保护, 2010, 36(2):66-70.
[12] Sareh B R, Esmat M M. Efficacy of Bacillus thuringiensis cry14 toxin against root knot nematode, Meloidogyne javanica[J]. Plant Protect Science, 2015, 51(1):46-51.
[13] Eva V A R, José R P G, Víctor M H V, et al. In vitro susceptibility of Varroa destructor and Apis mellifera to native strains of Bacillus thuringiensis[J]. Apidologie, 2014, 45(6):707-718.
[14] Sambrook J, Fritsch E F, Maniatis T, et al. Molecular cloning//A Laboratory Manual[M]. NY:Cold Spring Harbor Laboratory, 1989.
[15] 苏旭东. 苏云金芽胞杆菌菌株的分离和cry基因的鉴定[D]. 保定:河北农业大学, 2005.
[16] 宋福平, 张杰, 黄大昉, 等. 苏云金芽胞杆菌cry基因PCR-RFLP鉴定体系的建立[J]. 中国农业科学, 1998, 31(3):13-18.
[17] Narva K E, Payne J M, Schwab G E, et al. Novel Bacillus thuringiensis microbes active against nemotodes and genes encoding novel nemotodes active toxins cloned from Bt isolates[P]. Patent for an Invention, 1991, EP 19910305047.
[18] http://www.lifesci.sussex.ac.uk/home/Neil_Crickmore/Bt/[DB].
[19] 秦悦. 小菜蛾对Bt杀虫蛋白Cry1Ac抗性的分子机制研究[D]. 北京:中国农业科学院, 2011.
[20] 曹雪艳, 王会利, 张杰, 等. 9种杀虫剂对中华大刀螂幼虫的毒性[J]. 生态毒理学报, 2013, 8(3):435-441.
[21] 靳婷婷, 王振营, 何康来. 亚洲玉米螟中肠类钙粘蛋白Cry1A毒素结合区对Cry1Ac毒素杀虫活性的影响[J]. 中国生物防治学报, 2015, 31(1):41-47.

椰子织蛾幼虫致病Bt菌的鉴定与杀虫活性测定

Identification of Bacillus thuringiensis from Opisina arenosella and Determination of Its Insecticidal Bioactivity

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]	赵劲捷, 王帅, 范海燕, 赵迪, 刘晓宇, 朱晓峰, 王媛媛, 段玉玺, 陈立杰. 防治南方根结线虫的根瘤内生细菌的筛选及鉴定[J]. 中国生物防治学报, 2020, 36(5): 811-820.
[2]	吴琦琦, 吕宝乾, 曹凤勤, 吴晓霜, 卢辉, 唐继洪, 陆庆志, 张志扬. 椰子织蛾蛹寄生蜂——金刚钻大腿小蜂的生物学特性[J]. 中国生物防治学报, 2020, 36(4): 534-538.
[3]	罗梅, 罗玉霖, 陈沫冰, 舒永馨, 陈欣瑜, 董章勇. 拟康宁木霉Tk1的分离鉴定、拮抗作用及其生物学特性[J]. 中国生物防治学报, 2020, 36(4): 581-586.
[4]	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.
[5]	孙旺旺, 闫丽, 陈昌龙, 田宇, 李晓颖, 陈杰, 谢华. 生菜软腐和菌核病拮抗菌贝莱斯芽胞杆菌BPC6鉴定与防效[J]. 中国生物防治学报, 2020, 36(2): 231-240.
[6]	季冠宁, 汪志鹏, 焦加国, 胡锋, 李辉信. 枯草芽胞杆菌M29产抑菌物质培养条件的优化及抑菌物质组成初探[J]. 中国生物防治学报, 2019, 35(6): 930-939.
[7]	千慧敏, 文艺, 赵辉, 倪云霞, 刘新涛, 邱睿, 李小杰, 赵新贝, 李淑君, 刘红彦. 烟草黑胫病和根黑腐病生防假单胞杆菌的筛选与鉴定[J]. 中国生物防治学报, 2019, 35(6): 940-948.
[8]	彭国雄, 张淑玲, 夏玉先. 杀虫真菌对草地贪夜蛾不同虫态的室内活性[J]. 中国生物防治学报, 2019, 35(5): 729-734.
[9]	彭国雄, 张淑玲, 张维, 夏玉先. 杀虫真菌与苏云金芽胞杆菌对草地贪夜蛾的联合室内杀虫活性研究[J]. 中国生物防治学报, 2019, 35(5): 735-740.
[10]	姚锦爱, 黄鹏, 陈峰, 黄敏敏, 余德亿. 建兰炭疽病拮抗放线菌的筛选及防治效果[J]. 中国生物防治学报, 2019, 35(5): 805-812.
[11]	圆圆, 张虹, 苏道拉呼, 峥嵘, 石玲玲, 吴慧玲. 抑制黄瓜枯萎病的链霉菌S-101及其生防作用研究[J]. 中国生物防治学报, 2019, 35(5): 813-820.
[12]	赵曼, 汤金荣, 牛琳琳, 陈琳, 梁革梅. 不同Bt杀虫蛋白通过食物链对大草蛉的影响[J]. 中国生物防治学报, 2019, 35(4): 557-562.
[13]	王帆, 向剑, 胡军华, 李春峰, 陈若霓, 陈仕江, 卿玉玲, 秦少容. 一株高致病力赭绿青霉的生物学特性及杀虫活性[J]. 中国生物防治学报, 2019, 35(4): 576-585.
[14]	王燕, 王春伟, 王琳, 刘倩如, 董田, 侯普兴, 张曦倩, 王美琴, 张作刚, 王建明. 梨灰霉病菌拮抗细菌Pseudomonas extremorientalis的鉴定、发酵条件优化及防效评价[J]. 中国生物防治学报, 2019, 35(3): 437-448.
[15]	宋健, 曹伟平, 张晓, 杜立新. 对葱黄寡毛跳甲高毒力Bt菌株的鉴定及效果评价[J]. 中国生物防治学报, 2019, 35(2): 197-202.