感染油茶象幼虫的高致病力金龟子绿僵菌菌株筛选

doi:10.11707/j.1001-7488.20150807

摘要/Abstract

摘要：

[目的] 筛选对油茶象幼虫具有高致病力的绿僵菌菌株,为防治油茶象幼虫提供高效的生防菌。[方法] 在测定7个金龟子绿僵菌菌株的生长速度和产孢量的基础上,采用浸渍法和孢子拌土法对油茶象幼虫进行致病力测定,并以土表喷菌液的方式进一步研究优良菌株对土壤中油茶象幼虫的感染效果。[结果] FJMa201101和FJMa201205 两个菌株产孢量最高; 不同绿僵菌菌株对油茶象幼虫均有一定的致病力,10⁷孢子·mL^-1的孢悬液浸渍法接种后第13天和10⁵孢子·g^-1土的孢子拌土法接种后第9天,油茶象幼虫的死亡率均达到100%,LT₅₀分别在1.65~4.26天和1.96~3.51天之间; FJMa201101和FJMa201205 两个菌株对油茶象幼虫的致病力最强。FJMa201101菌株浸渍法接种LT₅₀为1.65天,感染率为86.6%; 孢子拌土法LT₅₀为2.11天,感染率为94.5%。FJMa201205菌株浸渍法接种LT₅₀为1.71天,感染率为91.1%; 孢子拌土法LT₅₀为1.96天,感染率为88.9%。林间土表喷菌液的方式,FJMa201101和FJMa201205 两个菌株之间的感染效果差异不显著,但先喷菌再引入试虫较引入试虫后再喷菌的感染效果更好。[结论] FJMa201101和FJMa201205 两个金龟子绿僵菌菌株可以作为防治油茶象幼虫的优良菌株在生产上应用。

关键词: 油茶象, 金龟子绿僵菌, 致病力, 浸渍法, 孢子拌土法, 蛀果害虫

Abstract:

[Objective] Curculio chinensis (Coleoptera: Curculionidae) is one of the most serious pests of oil-tea Camellia fruits.Screening of Metarhizium anisopliae strains with high virulence against this pest is important for using fungi as biocontrol agent to suppress this pest.[Method] Colony growth and sporulation of 7 strains of M. anisopliae were measured and their virulence against C. chinensis larvae were investigated in laboratory by using two inoculating methods ( Being dipped on larvae with conidia suspension of 10⁷ spores·mL^-1and being mixed in soil with 10⁵spores·g^-1). The infection effects in field were evaluated through spraying conidia suspension of two superior strains. [Result] Bioassay results showed that larvae of C. chinensis were susceptible to different M. anisopliae strains. The larval mortalities reached 100% on the 13th day after inoculation using dipping inoculative method and the 9th day for conidia-soil mixing inoculative method. The median lethal time (LT₅₀) of dipping inoculative method was 1.65 days to 4.26 days, and that of conidia-soil mixing inoculative method was 1.96 days to 3.51 days. The FJMa201101 and FJMa201205 strains of M. anisopliae were the most virulent against larvae of C. chinensis. When larvae inoculated with FJMa201101, the LT₅₀ was 1.65 days, and the infection rate was 86.6% when using dipping inoculative method, while the LT₅₀ was 2.11 days and the infection rate was 94.5% respectively for conidia-soil mixing inoculative method. When inoculated with FJMa201205, the LT₅₀ was 1.71 days with the infection rate of 91.1% by using dipping inoculative method, whilst it was 1.96 days with the infection rate of 88.9% with conidia-soil mixing inoculative method. In field test, there was no significant difference in infection rate between the two superior strains, but the infection rates of larvae introduced to soil after spraying conidia suspension were significantly higher than those introduced before spraying. Meanwhile, the two strains were excellent at growth performance. [Conclusion] Therefore, the two strains of M. anisopliae, FJMa201101 and FJMa201205, could be the promising candidate in controlling C. chinensis larvae.

Key words: Curculio chinensis, Metarhizium anisopliae, virulence, dipping inoculative method, conidia-soil mixing inoculative method, pest of oil-tea Camellia fruit

中图分类号:

S763.306

何学友, 蔡守平, 杜月飞, 陈德兰, 黄金水, 李孔泉. 感染油茶象幼虫的高致病力金龟子绿僵菌菌株筛选[J]. 林业科学, 2015, 51(8): 52-59.

He Xueyou, Cai Shouping, Du Yuefei, Chen Delan, Huang Jinshui, Li Kongquan. Screening of Metarhizium anisopliae Strain with High Virulence against Larvae of Curculio chinensis (Coleoptera: Curculionidae)[J]. Scientia Silvae Sinicae, 2015, 51(8): 52-59.

参考文献

蔡守平, 何学友, 李志真, 等. 2011. 油茶象危害油茶果实的初步研究. 福建林业科技, 38(2): 14-16. (Cai S P, He X Y, Li Z Z,et al. 2011. Study on damage of Curculio chinensis on Camellia oleifera fruit. Journal of Fujian Forestry Science and Technology, 38(2): 14-16.[in Chinese])
陈川, 唐周怀, 郭小侠, 等. 2009. 金龟子绿僵菌对华北大黑鳃金龟幼虫的杀虫活性研究. 中国农学通报, 25(4): 208-211. (Chen C, Tang Z H, Guo X X,et al. 2009. The pathogenicity of an isolate of the fungus Metarhizium anisopliae against the larvae of Holotrichia oblita (Fald.). Chinese Agricultural Science Bulletin, 25(4): 208-211.[in Chinese])
陈祝安, 谢佩华, 黄基荣, 等. 1995. 绿僵菌对暗黑金龟蛴螬的室内致病力测定. 中国生物防治, 11(2): 54-55. (Chen Z A, Xie P H, Huang J R,et al. 1995. Infectivity of Metarhizium anisopliae against the mulberry brown chafer Holotrichia parallela. Chinese Journal of Biological Control, 11(2): 54-55.[in Chinese])
董辉, 张泽华, 高松, 等. 2011. 绿僵菌治蝗对草原昆虫生物多样性的影响. 沈阳农业大学学报, 42 (1):37-40. (Dong H, Zhang Z H, Gao S,et al. 2011. The effect on grassland insects biodiversity of Metarhizium anisopliae controlling grasshopper. Journal of Shenyang Agricultural University, 42 (1):37-40.[in Chinese])
樊美珍, 李增智. 1996. 绿僵菌在土壤中的延续及控制桃小食心虫的潜力. 应用生态学报, 7(1): 49-55. (Fan M Z, Li Z Z. 1996. Persistence of Metarhizium anisopliae in soil and Its potential in controlling Carposina nipponensis. Chinese Journal of Applied Ecology, 7(1): 49-55.[in Chinese])
何立红, 李志文, 刘劲军, 等. 2014. 油茶象危害与小果油茶果实特征的相关性. 林业科学, 50(12): 151-155. (He L H, Li Z W, Liu J J, et al. 2014. Correlation between damage of Curculio chinensis and fruit traits of Camellia meioearpa. Scientia Silvae Sinicae, 50(12): 151-155.[in Chinese])
何学友, 蔡守平, 黄金水, 等. 2011. 应用绿僵菌和白僵菌林间防治木麻黄毒蛾的初步研究. 福建林业科技, 38(2): 10-13. (He X Y, Cai S P, Huang J S, et al. 2011. Preliminary study on controlling Lymantria xylina in Casuarina forest by using Metarhizium anisopliae and Beauveria bassiana. Journal of Fujian Forestry Science and Technology, 38(2): 10-13.[in Chinese])
何学友, 陈顺立, 黄金水. 2005. 感染松墨天牛的金龟子绿僵菌菌株的初步筛选. 昆虫学报, 48(6): 975-981. (He X Y, Chen S L, Huang J S,et al. 2005. Preliminary screening of virulent strains of Metarhizium anisopliae against Monochamus alternatus. Acta Entomologica Sinica, 48(6): 975-981.[in Chinese])
蒋三俊. 2009. 油茶象鼻虫的防治. 特种经济动植物, (8): 54. (Jiang S J. 2009. Control ofCurculio chinensis. Special Economic Animal and Plant, (8): 54.[in Chinese])
姜晓装. 2001. 油茶病虫害的防治措施. 林业科技开发, 15(5): 35-37. (Jiang X Z. 2001. Control strategy and technique against pests and diseases of Camellia oleifera. China Forestry Science and Technology, 15(5): 35-37.[in Chinese])
焦玉海. 2013. 全国油茶种植面积已达383万公顷. 林业与生态, (12): 46. (Jiao Y H. 2013. Cultivated area of Camellia oleifera reached 383 hm² in China. Forestry and Ecology, (12): 46.[in Chinese])
刘爱英. 1991. 中国虫生真菌的研究和应用. 北京: 中国农业科技出版社, 159-164. (Liu A Y. 1991. Research and application of entomogenous fungus in China. Beijing: China Agriculture Science and Technology Press, 159-164.[in Chinese])
陆永跃, 梁广文. 2004. 绿僵菌防治香蕉假茎象甲的使用技术. 华南农业大学学报, 25(3): 70-72. (Lu Y Y, Liang G W. 2004. Control efficience of Metarhizium anisopliae on the banana pseudostem weevil (Odoiporus longicollis). Journal of South China Agricultural University, 25(3): 70-72.[in Chinese])
宋漳, 江英成. 2001. 绿僵菌防治第1代马尾松毛虫的研究. 应用与环境生物学报, 7(1): 604-608. (Song Z, Jiang Y C. 2001. Application of Metarhizium anisopliae to control the first generation of Dendrolimus punctatus. Chinese Journal of Applied and Environmental Biology, 7(1): 604-608.[in Chinese])
唐启义, 冯明光. 2002. 实用统计分析及其DPS数据处理系统. 北京: 科学出版社. (Tang Q Y, Feng M G, 2002. DPS Data processing system for practical statistics. Beijing: Science Press.[in Chinese])
童应华, 陈顺立, 林强. 2009. 绿僵菌超低容量油剂防治萧氏松茎象试验研究. 福建林学院学报, 29(1): 33-36. (Tong Y H, Chen S L, Lin Q. 2009. Experiment study on controlling Hylobitelus xiaoi with oil formulation of Metarhizium anisopliae by ultra-low-volume spraying. Journal of Fujian College of Forestry, 29(1): 33-36.[in Chinese])
王宝辉, 郑建伟, 黄大庄, 等. 2009. 绿僵菌MS01菌株的生物学特性及在不同温湿度下对光肩星天牛幼虫的致病力. 林业科学, 45(9): 151-162. (Wang B H, Zheng J W, Huang D Z,et al. 2009. Biological characteristics of Metarhizium and effects of temperature and humidity on the pathogenicity against Anoplophora glabripennis larvae. Scientia Silvae Sinicae, 45(9): 151-162.[in Chinese])
吴青, 曾玲, 徐大高, 等. 2006. 感染椰心叶甲绿僵菌菌株的筛选. 华南农业大学学报, 27(2): 32-34. (Wu Q, Zeng L, Xu D G,et al. 2006. Screening of Metarhizium anisopliae strains for controlling Brontispa longissima. Journal of South China Agricultural University, 27(2): 32-34.[in Chinese])
肖育贵,郭亨孝. 2000. 绿僵菌防治鞭角华扁叶蜂试验研究. 四川林业科技, 21(3): 10-13.
Xiao Y G, Guo H X. 2000. A study on Metarhizium anisopliae for control of Chinolyda flagelli. Journal of Sichuan Forestry Science and Technology, 21(3): 10-13.[in Chinese])
赵文琴, 樊美珍, 蔡守平, 等. 2005. 不同绿僵菌、白僵菌菌株对铜绿丽金龟幼虫的毒力生物测定. 生物学杂志, 22(5): 43-45. (Zhao W Q, Fan M Z, Cai S P,et al. 2005. The virulence test of different strains of Metarhizium and Beauveria against Anomala corpulenta mlotsch. Journal of Biology, 22(5): 43-45.[in Chinese])
张雯龙, 谭志琼. 2012. 不同来源绿僵菌菌株对椰心叶甲的毒力测定. 云南农业大学学报, 27(1): 34-38. (Zhang W L, Tan Z Q. 2012. The virulence test of different strains of Metarhizium anisopliae against Brontispa longissima. Journal of Yunnan Agricultural University, 27(1): 34-38.[in Chinese])
张晶, 覃伟权, 阎伟, 等. 2012. 金龟子绿僵菌对红棕象甲的室内致病力测定. 热带作物学报, 33(5):899-905. (Zhang J, Qin W Q, Yan W,et al. 2012. Detection of pathoqenicity of Meatarhiziums against Rhynchophorus ferrugineus in laboratory. Chinese Journal of Tropical Crops, 33(5):899-905.[in Chinese])
周燚, 王中康, 喻子牛. 2006. 微生物农药研发与应用. 北京: 化学工业出版社. (Zhou Y, Wang Z K, Yu Z N. 2006. Development and Application of Microbial Pesticide. Beijing: Chemical Industry Press.[in Chinese])
庄乾营, 刘同金, 赵维, 等. 2010. 金龟子绿僵菌防治苹果桃小食心虫和草地蝗虫的效果. 山东农业科学, 45(10): 92-93. (Zuang Q Y, Liu T J, Zhao W,et al. 2010. Effect of control Carposina sasakii and Eacromius coerulipes using Metarhizium anisopliae. Shandong Agricultural Sciences, 45(10): 92-93.[in Chinese])
Ansari M A, Vestergaard S, Tirry L,et al. 2004. Selection of a highly virulent fungal isolate, Metarhizium anisopliae CLO 53, for controlling Hoplia philanthus. Journal of Invertebrate Pathology, 85(2): 89-96.
Flores A G, De La Rosa W, Rojas J C, et al. 2002. Evaluation of Beauveria bassiana and Metarhizium anisopliae (mitosporic) against species of the "white grub complex" in the South of Mexico. Southwestern Entomologist, 27(1): 73-83.
Mazodze R, Zvoutete P. 1999. Efficacy of Metarhizium anisopliae against Heteronychus licas (Scarabaedae: Dynastinae) in sugarcane in Zimbabwe. Crop Protect, 18(9): 571-575.
Milner R J, Samson P, Morton R. 2003. Persistence of conidia of Metarhizium anisopliae in sugarcane fields: effect of isolate and formulation on persistence over 3.5 years. Biocontrol Science and Technology, 13(5): 507-516.
Roberts D W, Leger R J S. 2004.Metarhizium spp., cosmopolitan insect-pathogenic fungi: mycological aspects. Advances in Applied Microbiology, 54: 1-70.

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