三株球孢白僵菌的分离鉴定与其对粘虫的室内毒力

doi:10.16409/j.cnki.2095-039x.2022.03.006

摘要/Abstract

摘要： 通过黄粉虫虫饵法从河北省采集的土样中分离得到三株昆虫病原真菌，结合形态学特征和rDNA序列分析对菌株进行了鉴定，结果表明：三株均为球孢白僵菌Beauveria bassiana，编号依次为Z-G2-5、M-G4、M-G3。通过生测发现3株球孢白僵菌菌株均能有效感染粘虫幼虫，三株球孢白僵菌对粘虫3龄幼虫的毒力不同：在5×10⁸孢子/mL剂量下，菌株Z-G2-5第6 d时对粘虫的累计校正死亡率达到96.77%，LT₅₀为3.02 d，LC₅₀为4.95×10⁶个孢子/mL。而菌株M-G3、M-G4的累计死亡率分别为66.67%、71.11%；LT₅₀分别为4.89 d、3.97 d；LC₅₀分别为1.18×10⁸个孢子/mL、1.13×10⁷个孢子/mL。上述结果表明，相比其他两株菌，菌株Z-G2-5对粘虫的致病力更强，致死速度更快，具有较大的应用价值，可作为粘虫幼虫生物防治的候选菌株。

关键词: 球孢白僵菌, 粘虫, 致病力, 生物防治, 鉴定

Abstract: Three entomopathogenic fungi strains were isolated from soil samples collected from Hebei Province using Tenebrio molitor larvae as bait and were identified through morphological characteristics and rDNA sequence analysis. Results showed that the three strains were Beauveria bassiana, named Z-G2-5, M-G4 and M-G3, respectively. Bioassay showed that all the three B. bassiana strains could infect the larvae of Mythimna separata effectively, but showed varying virulence to the 3^rd instar larvae of M. separata. At 5×10⁸ spores/mL, the strain Z-G2-5 caused a peak cumulative corrected mortality of 96.77% at the 6^th day post infection, with a LT₅₀ of 3.02 d, a LC₅₀ of 4.95×10⁶ spores/mL. At the same concentration, the cumulative mortality with M-G3 and M-G4 strains were only 66.67% and 71.11%, respectively, the LT₅₀ values were 4.89 d and 3.97 d, respectively, and the LC₅₀ values were 1.18×10⁸ spores/mL and 1.13×10⁷ spores/mL, respectively. The results indicated that, compared with the other two strains, the strain Z-G2-5 had stronger pathogenicity and faster lethal rate to armyworm larvae, and it can be used as a candidate strain with potential application for biological control of M. separata.

Key words: Beauveria bassiana, Mythimna separata, pathogenicity, biological control, identification

中图分类号:

S476.1

程茵, 郑吉阳, 王敦. 三株球孢白僵菌的分离鉴定与其对粘虫的室内毒力[J]. 中国生物防治学报, 2022, 38(2): 521-530.

CHENG Yin, ZHENG Jiyang, WANG Dun. Isolation and Identification of Three Beauveria bassiana Isolates and Their Virulence against Mythimna separata under Laboratory Condition[J]. Chinese Journal of Biological Control, 2022, 38(2): 521-530.

参考文献

[1] 姜玉英, 李春广, 曾娟, 等. 我国粘虫发生概况: 60年回顾[J]. 应用昆虫学报, 2014, 51(4): 890-898.
[2] Sharma H C, Sullivan D J, Bhatnagar V S. Population dynamics and natural mortality factors of the Oriental armyworm, Mythimna separata[J]. Crop Protection, 2002, 21(9): 721-732.
[3] 时春利. 北票地区玉米粘虫综合防治措施[J]. 现代农业, 2018, 507(9): 25.
[4] 张海龙. 玉米粘虫防治技术要点[J]. 农业与技术, 2019, 39(11): 105-106.
[5] 刘杰, 杨俊杰, 张智, 等. 2018年我国黏虫发生特点分析[J]. 植物保护, 2020, 46(1): 229-233.
[6] Wang J D, Chen L F, Wang Y R, et al. Silence of ryanodine receptor gene decreases susceptibility to chlorantraniliprole in the oriental armyworm, Mythimna separata Walker[J]. Pesticide Biochemistry and Physiology, 2018, 148: 34-41.
[7] Chen L F, Lin D J, Zhang J S, et al. Molecular cloning, characterization and functional analysis of GluCl from the oriental armyworm, Mythimna separata Walker[J]. Pesticide Biochemistry and Physiology, 2019, 156: 56-62.
[8] 江幸福, 张蕾, 程云霞, 等. 我国粘虫研究现状及发展趋势[J]. 应用昆虫学报, 2014, 51(4): 881-889.
[9] 刘杰, 姜玉英, 曾娟. 2013年我国黏虫发生特点分析[J]. 植物保护, 2015, 41(3): 131-137.
[10] 张云慧, 张智, 姜玉英, 等. 2012年三代黏虫大发生原因初步分析[J]. 植物保护, 2012, 38(5): 1-8.
[11] 潘蕾, 吴秋琳, 陈晓, 等. 华北三代粘虫大发生虫源的形成[J]. 应用昆虫学报, 2014, 51(4): 958-973.
[12] Roditakis E, Vasakis E, Garcia-Vidal L, et al. A four-year survey on insecticide resistance and likelihood of chemical control failure for tomato leaf miner Tuta absoluta in the European/Asian region[J]. Journal of Pest Science, 2018, 91: 421-435.
[13] 阙生全, 喻爱林, 刘亚军, 等. 白僵菌应用研究进展[J]. 中国森林病虫, 2019, 38(2): 29-35.
[14] 于健, 查萌, 郑梦君, 等. 白僵菌和绿僵菌对棉铃虫的室内防控效果评价[J]. 新疆农业科学, 2020, 57(4): 608-615.
[15] Zimmermann G. Review on safety of the entomopathogenic fungi Beauveria bassiana and Beauveria brongniartii[J]. Biocontrol Science and Technology, 2007, 17(5): 553-596.
[16] 黄少华, 张宝鑫, 李敦松, 等. 珠江三角洲甜玉米螟害防治试验初报[J]. 广东农业科学, 2004(1): 44-46.
[17] 刘晓晓, 王星冉, 王义勋, 等. 马尾松毛虫高毒力球孢白僵菌菌株的生物学特性[J]. 南方农业学报, 2017, 48(6): 1019-1023.
[18] 徐超民, 李霜, 孟祥晨, 等. 不同浓度增效剂对金龟子绿僵菌、黄绿绿僵菌和球孢白僵菌产孢的影响[J].中国生物防治学报, 2020, 5(4): 1-10.
[19] 陈斌, 李正跃, 桂富荣. 绿僵菌对草坪灰翅贪夜蛾和粘虫的毒力及田间防治效果[J]. 植物保护, 2004(1): 32-36.
[20] 张琛, 陈思博, 吴晨源, 等. 粘虫高毒力白僵菌菌株筛选及其感菌后虫体内保护酶活性的变化[J].核农学报, 2020, 34(12): 2701-2707.
[21] 蔡守平. 绿僵菌MaFZ-13对油桐鹰尺蠖致病力的时间-剂量-死亡率模型分析[J]. 中国森林病虫, 2020, 39(3): 1-4.
[22] 贾春生, 由士江, 高文韬. 利用黄粉虫分离土壤昆虫病原真菌[J]. 昆虫知识, 2006(2): 260-261, 280.
[23] Abolfazl M. 中国西部森林昆虫病原真菌多样性与致病力研究[D]. 杨凌: 西北农林科技大学, 2018.
[24] Bischoff J F, Rehner S A, Humber R A. A multilocus phylogeny of the Metarhizium anisopliae lineage[J]. Mycologia, 2009, 101(4): 512-530.
[25] White T J, Bruns T, Lee S. Analysis of phylogenetic relationships by amplification and direct sequencing of ribosomal genes//[M]. PCR Protocols: A Guide to Methods and Applications[J]. New York Academic, 1990, 315-322.
[26] Rehner S A, Posada F, Buckley E P, et al. Phylogenetic origins of African and Neotropical Beauveria bassiana s.l. pathogens of the coffee berry borer, Hypothenemus hampei[J]. Journal of Invertebrate Pathology, 2006, 93(1): 11-21.
[27] Kumar S, Tamura K, Nei M. MEGA3: Integrated software for molecular evolutionary genetics analysis and sequence alignment[J]. Briefings in Bioinformatics, 2004, 5(2): 150-163.
[28] 何学友, 潘爱芳, 蔡守平, 等. 金龟子绿僵菌FJMa2014-0501菌株对缀叶丛螟的致病力[J]. 福建林业科技, 2018, 45(2): 1-5.
[29] 吴志鹏, 童应华. 球孢白僵菌和金龟子绿僵菌对红火蚁工蚁的致病力测定[J]. 森林与环境学报, 2020, 40(1): 99-105.
[30] Vandenberg J D. Standardized bioassay and screening of Beauveria bassiana and Paecilomyces fumosoroseus against the Russian wheat aphid (Homoptera: Aphididae)[J]. Journal of Economic Entomology, 1996, 89(6): 1418-1423.
[31] 邓嘉茹, 孙龙华, 毕可可, 等. 埃及吹绵蚧昆虫病原真菌的筛选、鉴定及其致病性测定[J]. 植物保护学报, 2020, 47(1): 53-64.
[32] 庄宝龙, 赵薇, 裴海英, 等. 白僵菌菌株分离培养及对四种重要害虫的致病力[J]. 应用昆虫学报, 2020, 57(6): 1417-1426.
[33] Mandira K B. 白僵菌、绿僵菌和棒束孢对两种昆虫的毒力与病理学[D]. 杨凌: 西北农林科技大学, 2018.
[34] McClatchie G V, Moore D, Bateman R P, et al. Effects of temperature on the viability of the conidia of Metarhizium flavoviride in oil formulations[J]. Mycological Research, 1994, 98(7): 749-756.
[35] Abdulhai M, El-bouhssini M, Jamal M, et al. Beauveria bassiana characterization and efficacy vs. Sunn pest, Eurygaster integriceps Puton (Hemiptera: Scutelleridae)[J]. Pakistan Journal of Biological Sciences, 2010, 13(21): 1052-1056.
[36] Hiromori H, Yaginuma D, Kajino K, et al. The effects of temperature on the insecticidal activity of Beauveria amorpha to Heptophylla picea[J]. Applied Entomology and Zoology, 2004, 39(3): 389-392.
[37] Alali S, Mereghetti V, Faoro F, et al. Thermotolerant isolates of Beauveria bassiana as potential control agent of insect pest in subtropical climates[J]. PLoS ONE, 2019, 14(2): e0211457.
[38] Hegedus D D, Khachatourians G G. The effects of temperature on the pathogenicity of heat-sensitive mutants of the entomopathogenic fungus, Beauveria bassiana, toward the migratory grasshopper, Melanoplus sanguinipes[J]. Journal of Invertebrate Pathology, 1996, 68(2): 160-165.
[39] 王成树, 王四宝, 李增智, 等. 球孢白僵菌种群野生菌株异核现象的分子验证[J]. 菌物系统, 2001(4): 475-481.
[40] 张璐璐. 防治西花蓟马的球孢白僵菌液固双相发酵条件优化[D]. 北京:中国农业科学院, 2016.