Screening of Insecticidal Activity Protein against Holotrichia parallela Adults from Bacillus thuringiensis

doi:10.16409/j.cnki.2095-039x.2018.04.008

Abstract

Abstract: Holotrichia parallela is an important agricultural pests, damaging several crops at both the adult and larval stages. To screen Bacillus thuringiensis protein of high insecticidal activity a bioassay method of the Bt protein against H. parallela adults was established by feeding on the adults with Ulmus pumila L leaves soaked in Bt proteins. Six Cry8 full-length crystal proteins and trypsin-digested proteins were prepared and used in the insecticidal activity bioassay. Cry8Ga1 protoxin and digested protein showed high insecticidal activity in the primary test, with 7-day corrected mortality rates of 100% and 55%, respectively, at the concentration of 1.0 mg/mL. Further, the median lethal concentration (LC₅₀) values were determined to be 0.387 mg/mL (95% CL 0.108-0.746) for Cry8Ga1 protoxin and 0.702 mg/mL (95% CL 0.150-2.664) for digested Cry8Ga1 protein. When fed with Cry8Ga1 protoxin and digested protein, H. parallela adults exhibited obvious pathological changes at the organelles in midgut cells. The Bt protein showing high insecticidal activity against H. parallela adults is a new approach for the biological control of Scarabaeoideas.

Key words: Bacillus thuringiensis, Cry8Ga1 protein, Holotrichia parallela adults, insecticidal activity

CLC Number:

S476.1

HUANG Shizhi, SHU Changlong, LIU Chunqin, DING Xuezhi, ZHANG Jie. Screening of Insecticidal Activity Protein against Holotrichia parallela Adults from Bacillus thuringiensis[J]. journal1, 2018, 34(4): 546-552.

References

[1] 中华人民共和国农业部. 中国农业统计资料. 2013[M]. 北京:中国农业出版社, 2014.
[2] 张美翠, 尹姣, 李克斌, 等. 地下害虫蛴螬的发生与防治研究进展[J]. 中国植保导刊, 2014, 34(10):20-28.
[3] Hugo A, Hortensia C R, Miguel A M. Record of scarabaeoidea larvae and adults associated with Amaranthus hypochondriacus and living fences[J]. Southwestern Entomologist, 2017, 41(3):675-680.
[4] 姚庆学, 张勇, 丁岩. 金龟子防治研究的回顾与展望[J]. 东北林业大学学报, 2003, 31(3):64-66.
[5] 鞠倩, 李晓, 姜晓静, 等. 3种金龟甲对寄主植物的行为反应研究[J]. 植物保护, 2014, 40(4):76-79.
[6] Jouzani G S, Valijanian E, Sharafi R. Bacillus thuringiensis:a successful insecticide with new environmental features and tidings[J]. Applied Microbiology Biotechnology, 2017, 101(7):2691-2711.
[7] Shu C, Liu R, Wang R, et al. Improving toxicity of Bacillus thuringiensis strain contains the cry8Ca gene specific to Anomala corpulenta larvae[J]. Current Microbiology, 2007, 55(6):492-496.
[8] Yu H, Zhang J, Huang D, et al. Characterization of Bacillus thuringiensis strain Bt185 toxic to the Asian cockchafer:Holotrichia parallela[J]. Current Microbiology, 2006, 53(1):13-17.
[9] Shu C, Yu H, Wang R, et al. Characterization of two novel cry8 genes from Bacillus thuringiensis strain BT185.[J]. Current Microbiology, 2009, 58(4):389-392.
[10] Shu C, Yan G, Wang R, et al. Characterization of a novel cry8 gene specific to Melolonthidae pests:Holotrichia oblita and Holotrichia parallela[J]. Applied Microbiology Biotechnology, 2009, 84(4):701-707.
[11] 束长龙, 宋福平, 王容燕, 等. 对鞘翅目害虫高效的苏云金芽胞杆菌cry8H基因、蛋白及其应用[P]. 中国发明专利, 2010, ZL200710120020.2.
[12] 闫贵欣, 束长龙, 张杰, 等. 对鞘翅目害虫高效的苏云金芽胞杆菌cry8Ⅰ基因、蛋白及其应用[P]. 中国发明专利, 2010, ZL 200810226214.5.
[13] Bi Y, Zhang Y, Shu C, et al. Genomic sequencing identifies novel Bacillus thuringiensis Vip1/Vip2 binary and Cry8 toxins that have high toxicity to Scarabaeoidea larvae[J]. Applied Microbiology Biotechnology, 2015, 99(2):753-760.
[14] Asano S, Yamashita C, Iizuka T, et al. A strain of Bacillus thuringiensis subsp. galleriae containing a novel cry8 gene highly toxic to Anomala cuprea (Coleoptera:Scarabaeidae)[J]. Biological Control, 2003, 28(2):191-196.
[15] Yamaguchi T, Sahara K, Bando H, et al. Discovery of a novel Bacillus thuringiensis Cry8D protein and the unique toxicity of the Cry8D-class proteins against scarab beetles[J]. Journal of Invertebrate Pathology, 2008, 99(3):257-262.
[16] Zhang Y, Zheng G, Tan J, et al. Cloning and characterization of a novel cry8Ab1 gene from Bacillus thuringiensis strain B-JJX with specific toxicity to scarabaeid (Coleoptera:Scarabaeidae) larvae[J]. Microbiological Research, 2013, 168(8):512-517.
[17] 茅洁瑜, 束长龙, 李克斌, 等. 对蛴螬有活性的苏云金芽胞杆菌菌株的筛选与基因鉴定[J]. 中国生物防治学报, 2011, 27(2):176-181.
[18] Zhou Z, Yang S, Shu C, et al. Comparison and optimization of the method for Cry1Ac protoxin preparation in HD73 strain[J]. Journal of Integrative Agriculture, 2015, 14(8):1598-1603.
[19] 萨姆布鲁克J, 拉塞尔D W著. 黄培堂译. 分子克隆实验指南(第3版)[M]. 北京:科学出版社, 2002.
[20] 张彦, 梁革梅, 张丽丽, 等. 棉铃虫幼虫取食Vip3Aa蛋白后的中肠组织病理变化[J]. 昆虫学报, 2012, 55(7):869-876.
[21] 魏红爽, 谭树乾, 曹雅忠, 等. 暗黑鳃金龟幼虫取食Bt菌株HD8E和HD8G后的中肠细胞病理变化[J]. 应用昆虫学报, 2017, 54(6):1015-1022.
[22] 李杨, 韩君, 于春雷, 等. 七种杀虫剂对暗黑鳃金龟成虫和幼虫的毒力及田间防控效果[J]. 植物保护学报, 2012, 39(2):147-152.
[23] 刘福顺, 王庆雷, 刘春琴, 等. 大黑蛴螬活动规律及对农作物幼苗的取食趋性研究[J]. 环境昆虫学报, 2014, 36(4):635-639.
[24] 朱秀蕾, 陆秀君, 刘子欢, 等. 六种杂草对华北大黑鳃金龟甲取食和繁殖的影响[J]. 植物保护学报, 2017, 44(2):351-352.
[25] Grochulski P, Masson L, Borisova S, et al. Bacillus thuringiensis CryIA(a) insecticidal toxin:crystal structure and channel formation[J]. Journal of Molecular Biology, 1995, 254(3):447-464.
[26] Bravo A, Gómez I, Conde J, et al. Oligomerization triggers binding of a Bacillus thuringiensis Cry1Ab pore-forming toxin to aminopeptidase N receptor leading to insertion into membrane microdomains[J]. Biochimica et Biophysica Acta (BBA) Biomembranes, 2004, 1667(1):38-46.
[27] Bravo A, Likitvivatanavong S, Gill S S, et al. Bacillus thuringiensis:A story of a successful bioinsecticide[J]. Insect Biochemistry Molecular Biology, 2011, 41(7):423-431.
[28] Zhang X, Candas M, Griko N B, et al. A mechanism of cell death involving an adenylyl cyclase/PKA signaling pathway is induced by the Cry1Ab toxin of Bacillus thuringiensis[J]. Proceedings of the National Academy of Sciences, 2006, 103(26):9897-9902.
[29] 彭琦, 周子珊, 张杰. 苏云金芽胞杆菌杀虫晶体蛋白研究进展[J]. 中国生物防治学报, 2015, 31(5):712-722.
[30] Shu C, Tan S, Yin J, et al. Assembling of Holotrichia parallela, (dark black chafer) midgut tissue transcriptome and identification of midgut proteins that bind to Cry8Ea toxin from Bacillus thuringiensis[J]. Applied Microbiology Biotechnology, 2015, 99(17):7209-7218.
[31] 张美翠, 尹姣, 曹雅忠, 等. 暗黑鳃金龟幼虫类钙粘蛋白基因的克隆及组织表达特异性分析[J]. 西北农业学报, 2015, 24(9):152-160.
[32] Zhao D, Guo W, Li S, et al. Identification of a new peritrophic membrane protein from larval Holotrichia parallela (Coleoptera:Motschulsky)[J]. Molecules, 2014, 19(11):17799-17809.