Abstract:Rhynchophorus ferrugineus is a major quarantine pest that damages the coconut trees (Cocos nucifera) and other palm plants (Arecaceae). In this study, the strain Yel-8 was isolated from the corpse of R. ferrugineus, which had insecticidal activity against the larvae of R. ferrugineus which was suffered from sepsis and identified by morphological identification, 16S rDNA, physiological and biochemical tests, and drug susceptibility analysis, respectively. The results showed that Yel-8 was in line with the characteristic of Yersinia entomophaga in L-arabinose, rhamnose, xylose, sucrose, aesculin, and cellobiose metabolism, whereas opposed to Yersinia only in sorbitol metabolism. Therefore, this strain was identified as a new strain of Yersinia genus. The bioassay of Yel-8 against R. ferrugineus was performed using feeding and injection methods, and the median lethal concentration (LC50) of Yel-8 against R. ferrugineus was 3.184×108 and 5.1295×106 CFU/mg, respectively. The chitinase genes Chi1 and Chi2 were cloned from the Ye1-8 strain and the proteins were expressed for bioassay. The results showed that the Chi1 and Chi2 proteins had no insecticidal activity against R. ferrugineus alone, thus suggesting the necessity of complete toxin complex for toxicity. Yel-8 exhibited insecticidal activity may provide an ideal material for the construction of the biophytes against R. ferrugineus.
[1]Anonymous.Data sheets on quarantine pests. [J].Eppo Bulletin, 2005, 35(3):361-362
[2]Arfaoui A, Adam L R, Bezzahou A, et al.Isolation and identification of cultivated bacteria associated with soybeans and their biocontrol activity against Phytophthora sojae.[J].BioControl, 2018, 1(7):1-11
[3]Busby J N, Landsberg M J, Simpson R M, et al.Structural analysis of Chi1 Chitinase from Yen-Tc: the multisubunit insecticidal ABC toxin complex of Yersinia entomophaga. [J].Journal of Molecular Biology, 2012, 415(2):359-371
[4]Carsolio C, Guti??rrez A, Jim??nez B, et al.Characterization of ech-42, a Trichoderma harzianum endochitinase gene expressed during mycoparasitism. [J].Proceedings of the National Academy of Sciences of the United States of America, 1994, 91(23):10903-10907
[5]Chernin L S, Fuente L D L, Sobolev V, et al.Molecular cloning, structural analysis, and expression in Escherichia coli of a chitinase gene from Enterobacter agglomerans.[J].Applied and Environmental Microbiology, 1997, 63(3):834-839
[6]Cover T L, Aber R C.Yersinia enterocolitica.[J].New England Journal of Medicine, 2013, 2(3):16-24
[7]Ewing W H, Ross A J, Brenner D J, et al.Yersinia ruckeri sp. nov., the redmouth (RM) bacterium. [J].International Journal of Systematic Bacteriology, 1978, 28(1):37-44
[8]Fouz B, Zarza C, Amaro C.First description of non-motile Yersinia ruckeri serovar I strains causing disease in rainbow trout, Oncorhynchus mykiss (Walbaum), cultured in Spain. [J].Journal of Fish Diseases, 2006, 29(6):339-339
[9]Guan Y, Zhang W, Deng M, et al.Significant enhancement of photobiological H2 evolution by carbonylcyanide m-chlorophenylhydrazone in the marine green alga Platymonas subcordiformis. [J].Biotechnology Letters, 2004, 26(13):1031-1035
[10]Hurst M R, Jones S A, Binglin T, et al.The main virulence determinant of Yersinia entomophaga MH96 is a broad-host-range toxin complex active against insects.[J].Journal of Bacteriology, 2011, 193(8):1966-1980
[11]Hurst M R H, Becher S A, Young S D, et al.Yersinia entomophaga sp. nov., isolated from the New Zealand grass grub Costelytra zealandica.[J].International Journal of Systematic & Evolutionary Microbiology, 2011, 61(4):844-849
[12]Hurst M R H, Koten C V, Jackson T A.Pathology of Yersinia entomophaga MH96 towards Costelytra zealandica (Coleoptera; Scarabaeidae) larvae.[J].Journal of Invertebrate Pathology, 2014, 115(1):102-102
[13]Kuranda M J, Robbins P W.Cloning and heterologous expression of glycosidase genes from Saccharomyces cerevisiae.[J].Proceedings of the National Academy of Sciences of the United States of America, 1987, 84(9):2585-2589
[14]Landsberg M J, Jones S A, Rothnagel R, et al.3D structure of the Yersinia entomophaga toxin complex and implications for insecticidal activity.[J].Proceedings of the National Academy of Sciences of the United States of America, 2011, 108(51):20544-20549
[15]Liang Y, Jing M, Tan L, et al.Isolation and identification of biocontrol strain and its effect on the control of potato late blight.[J].Journal of Northern Agriculture, 2017, :-
[16]Limón M C, Lora J M, García I, et al.Primary structure and expression pattern of the 33-kDa chitinase gene from the mycoparasitic fungus Trichoderma harzianum.[J].Current Genetics, 1995, 28(5):478-478
[17]Liu M, Cai Q X, Liu H Z, et al.Chitinolytic activities in Bacillus thuringiensis and their synergistic effects on larvicidal activity.[J].Journal of Applied Microbiology, 2010, 93(3):374-379
[18]Martin P a W, Shropshire A D S, Gundersen-Rindal D E, et al.2007. Chromobacterium subtsugae sp. nov. for control of insect pests. US 7244607 B2 [P].
[19]Morgan J A, Sergeant M, Ellis D, et al.Sequence analysis of insecticidal genes from Xenorhabdus nematophilus PMFI296.[J].Applied and Environmental Microbiology, 2001, 67(5):2062-2062
[20]Murroskontiainen A, Fredrikssonahomaa M, Korkeala H, et al.Yersinia nurmii sp. nov.[J].International Journal of Systematic & Evolutionary Microbiology, 2010, 61(10):2368-2372
[21]Paff J R, Triplett D A, Saari T N.Clinical and Laboratory Aspects of Yersinia pseudotuberculosis Infections, with a Report of Two Cases.[J].American Journal of Clinical Pathology, 1976, 66(1):101-110
[22]Peng L, Hou Y.Red Palm Weevil Rhynchophorus ferrugineus (Olivier).[J].Biological Invasions and Its Management in China, 2017, 11:245-256
[23]Perry R D, Fetherston J D.Yersinia pestis--etiologic agent of plague.[J].Clinical Microbiology Reviews, 1997, 10(1):35-66
[24]Sulakvelidze A.Yersiniae other than Y. enterocolitica, Y. pseudotuberculosis, and Y. pestis: the ignored species.[J].Microbes & Infection, 2000, 2(5):497-513
[25]Suzuki K, Taiyoji M, Sugawara N, et al.The third chitinase gene (chiC) of Serratia marcescens 2170 and the relationship of its product to other bacterial chitinases.[J].Biochemical Journal, 1999, 343(3):587-587
[26]Tamura K, Stecher G, Peterson D, et al.MEGA6: Molecular Evolutionary Genetics Analysis version 6.0.[J].Molecular Biology and Evolution, 2013, :2725-2729
[27]Tobback E, Decostere A, Hermans K, et al.Yersinia ruckeri infections in salmonid fish.[J].Journal of Fish Diseases, 2007, 30(5):257-257
[28]Wang S, Ala D S, Huang W, et al.Driving mosquito refractoriness to Plasmodium falciparum with engineered symbiotic bacteria.[J].Science, 2017, 357(3658):1399-1402
[29]Wang S, Ghosh A K, Bongio N, et al.Fighting malaria with engineered symbiotic bacteria from vector mosquitoes.[J].Proceedings of the National Academy of Sciences of the United States of America, 2012, 109(31):12734-12739
[30]Wiwat C, Thaithanun S, Pantuwatana S, et al.Toxicity of chitinase-producing Bacillus thuringiensis ssp. kurstaki HD-1 (G) toward Plutella xylostella.[J].Journal of Invertebrate Pathology, 2000, 76(4):270-277
[31]Woese C R, Fox G E.Phylogenetic structure of the prokaryotic domain: the primary kingdoms.[J].Proceedings of the National Academy of Sciences of the United States of America, 1977, 74(11):5088-5090
[32]崔玉军, 宋亚军, 杨瑞馥.鼠疫耶尔森氏菌的进化研究: 从系统发育学到系统发育基因组学[J].中国科学:生命科学, 2013, 1(1):23-30
[33]东秀珠, 蔡妙英.2001. 常见细菌系统鉴定手册 [M]. 北京: 科学出版社, 267-295. (Dong X Z, Cai M Y. 2001. Manual for identification of common bacterial systems [M]. Beijing: Science Press, 267-295.)
[34]范方玲, 汪开毓, 耿毅, 等.斑点叉尾(Ictalunes punctatus)源鲁氏耶尔森氏菌的分离鉴定及系统发育分析. [J].海洋与湖沼, 2010, 41(6):862-868
[35]费来玺, 何国协.耶尔森氏菌属研究近况. [J].地方病译丛, 1987, 1(4):1-6
[36]侯有明, 郭伟艺, 李猷, 等.2013. 一种红棕象甲半人工饲料及其制备方法. 中国, 103431160 A [P]. (Hou Y M, Guo W Y, Li Y, et al. 2013. A red palm weevil semi-artificial feed and its preparation method. China, 103431160 A [P].)
[37]姜海荣, 倪永清, 宋丽军, 等.新鲜哈密瓜汁中可培养细菌的分离鉴定及系统发育分析. [J].食品与生物技术学报, 2010, 29(3):426-431
[38]卢伟, 蔡峻, 陈月华.苏云金芽孢杆菌几丁质酶的研究进展.[J].微生物学通报, 2007, 34(1):143-147
[39]牟亚妮, 徐卓吟, 翁小倩, 等.铜绿假单胞菌BRC-CXG5菌株的分离鉴定及其对松墨天牛(Monochamus alternatus Hope)幼虫的杀虫活性.[J].生物技术进展, 2016, 6(1):41-46
[40]覃伟权, 赵辉, 韩超文.红棕象甲在海南发生为害规律及其防治.[J].热带农业科技, 2002, 25(4):29-30
[41]吴松青, 陈思琪, 陈小刚, 等.沙雷氏菌菌株BRC-CXG2的分离、鉴定及其粗提物对苏云金芽胞杆菌的增效作用.[J].农业生物技术学报, 2015, 23(11):1516-1523
[42]熊悦婷, 夏枫, 黄志成, 等.芽孢杆菌BRC-XYT1菌株的分离和鉴定及其对松墨天牛(Monochamus alternatus Hope)的杀虫活性.[J].生物技术进展, 2016, 6(1):47-51
[43]张令要, 岳木生, 袁平, 等.神农架原始森林高毒力苏云金芽孢杆菌菌株分离鉴定研究.[J].中国媒介生物学及控制杂志, 2001, 12(4):271-274