木贼镰刀菌的鉴定及其对桃蚜的致病性测定

doi:10.16409/j.cnki.2095-039x.2022.07.002

摘要/Abstract

摘要： 从罹病桃蚜虫尸上分离到了一株昆虫病原真菌，命名为JMF-01，本文旨在确定该菌株分类地位并探索其生防潜能。对罹病的桃蚜虫尸进行分离纯化，基于形态学观察和rDNA-ITS、RPB2、IGS序列分析，构建系统发育树对该菌株进行鉴定。采用浸叶浸虫法研究该菌株不同浓度条件下对桃蚜的致病力并进行温室防效测定。结果表明菌株JMF-01在PDA培养基上5 d后菌落直径58～60 mm，分生孢子镰刀形，3～7隔。菌株JMF-01对桃蚜具有较强致病力，处理桃蚜在7 d后累计校正死亡率和LC₅₀达到85%和9.87×10⁵ cfu/mL；孢子悬浮液最高浓度的温室防效在14 d时高于70%。该菌株的rDNA-ITS序列（MW404610）与木贼镰刀菌Fusarium equiseti（GenBank登录号：JF773657）相似度达100%，位于系统发育树的同一分支；RPB2、IGS序列分别与木贼镰刀菌（GenBank登录号：MK077112，KX583611）的相似度也达到99%以上，聚在系统发育树的同一分支。菌株JMF-01经鉴定为桃蚜的病原真菌木贼镰刀菌，具有生防潜力。

关键词: 桃蚜, 木贼镰刀菌, 昆虫病原真菌, 致病力

Abstract: A strain of entomopathogenic fungus was isolated from the corpse of Myzus persicae and designated as JMF-01. In this study, the author aims to determine the taxonomic status of the strain and explore its biocontrol potential. The strain was isolated and purified from the diseased peach aphid corpse. Based on morphological observation and rDNA-ITS, RPB2 and IGS sequence analysis, the phylogenetic tree was constructed to identify the strain. The pathogenicity of the strain to M. persicae under different spore suspension concentrations was measured by leaf dipping and insect dipping method and the control of the aphids by the strain was determined in greenhouse tests. The results showed that the colony diameter of JMF-01 was 58―60 mm at 5 days on PDA medium, conidia were sickle-shaped and separated by 3―7. Strain JMF-01 showed strong virulence to M. persicae and the cumulative corrected mortality and LC₅₀ reached 85% and 9.87×10⁵ cfu/mL at 7 days after treatment, respectively. The control efficiency of the aphids by the spore suspension at the highest concentration in greenhouse was higher than 70% at 14 days after treatment. The rDNA-ITS sequence (MW404610) of the strain was 100% consistent with that of Fusarium equiseti (GenBank accession number: JF773657), locating in the same branch of the phylogenetic tree; the similarity of RPB2 and IGS sequences with F. equiseti (GenBank accession number: MK077112, KX583611) also reached more than 99%, clustering in the same branch of the phylogenetic tree. In conclusion, the strain JMF-01 identified as F. equiseti is a virulent pathogen of M. persicae with high biocontrol potential.

Key words: Myzus persicae, Fusarium equiseti, entomopathogenic fungi, pathogenicity

中图分类号:

S476.1

赵雪怡, 柴军发, 张暄翊, 洪波, 贾彦霞. 木贼镰刀菌的鉴定及其对桃蚜的致病性测定[J]. 中国生物防治学报, 2023, 39(1): 69-76.

ZHAO Xueyi, CHAI Junfa, ZHANG Xuanyi, HONG Bo, JIA Yanxia. Identification of Fusarium equiseti and Its Pathogenicity to Myzus persicae[J]. Chinese Journal of Biological Control, 2023, 39(1): 69-76.

参考文献

[1] Mohammed A A, Hatcher P E. Combining entomopathogenic fungi and parasitoids to control the green peach aphid Myzus persicae[J]. Biological Control, 2017, 110:44-55.
[2] Bass C, Puinean A M, Zimmer C T, et al. The evolution of insecticide resistance in the peach potato aphid, Myzus persicae[J]. Insect Biochemistry and Molecular Biology, 2014, 51:41-51.
[3] Prince G, Chandler D. Susceptibility of Myzus persicae, Brevicoryne brassicae and Nasonovia ribisnigri to fungal biopesticides in laboratory and field experiments[J]. Insects, 2020, 11(1):55-70.
[4] 汤秋玲,马康生,高希武. 蔬菜蚜虫抗药性现状及抗性治理策略[J]. 植物保护, 2016, 42(6):11-20.
[5] 张帅. 2015年全国农业有害生物抗药性监测结果及科学用药建议[J]. 中国植保导刊, 2016, 36(3):61-65.
[6] Simon J, Peccoud J. Rapid evolution of aphid pests in agricultural environments[J]. Current Opinion in Insect Science, 2018, 26:17-24.
[7] 张平艳, 周小毛. 华南地区桃蚜对7种杀虫剂的抗药性研究[J]. 广东农业科学, 2014, 41(10):81-84.
[8] 王峥, 王秀梅, 阮长春. 两种蚜茧蜂在桃蚜上的繁殖特性及其控害潜能评估[J]. 吉林农业大学学报, 2020, 42(1):20-25.
[9] 田佳, 汝冰璐, 王颖, 等. 一株对桃蚜有高致病性球孢白僵菌的分离、筛选与鉴定[J]. 植物保护学报, 2018, 45(3):606-613.
[10] 王利军, 谭万忠, 罗华东, 等. 虫生真菌及其在害虫生物控制中的应用现状与展望[J]. 河南农业科学, 2010(4):119-125.
[11] 王记祥, 马良进. 虫生真菌在农林害虫生物防治中的应用[J]. 浙江林学院学报, 2009, 26(2):286-291.
[12] 亓兰达, 魏鹏霖, 张晗星, 等. 后基因组时代害虫生防真菌次级代谢产物的研究进展[J]. 中国科学:生命科学, 2020, 50(6):589-598.
[13] 王清海, 万平平, 黄玉杰, 等. 虫生真菌在害虫生物防治中的应用研究[J]. 山东科学, 2005, 18(4):37-41.
[14] 孙大平, 路茜, 王鹏, 等. 4种病原真菌对桃蚜和温室白粉虱的致病力[J]. 吉林农业大学学报, 2021, 43(6):664-672.
[15] 孟豪, 田晶, 付淑慧, 等. 玫烟色棒束孢与球孢白僵菌对桃蚜致病力对比[J]. 植物保护学报, 2014, 41(6):717-722.
[16] 贾廷祥, 刘传德, 吴桂本, 等. 小麦根腐镰刀菌鉴定及其生物学特性[J]. 植物保护学报, 1995(3):259-264.
[17] 闫文雪, 石延霞, 李盼亮, 等. 大白菜枯萎病病原镰刀菌种类的初步研究[J]. 植物病理学报, 2018, 48(5):587-593.
[18] Batta Y A. The first report on entomopathogenic effect of Fusarium avenaceum (Fries) Saccardo (Hypocreales, Ascomycota) against rice weevil (Sitophilus oryzae L.:Curculionidae, Coleoptera)[J]. Journal of Entomological and Acarological Research, 2012, 44(3):e11.
[19] 叶琪铭. 昆虫天敌——虫生镰刀菌.[J]. 昆虫天敌, 1989(1):5-9.
[20] 白逢彦, 陈庆涛. 中国部分昆虫上的镰刀菌[J]. 真菌学报, 1991, 10(2):120-128.
[21] 张挺峰, 王睿, 刘长仲. 一株侵染豌豆蚜的昆虫病原真菌的分离及鉴定[J]. 昆虫学报, 2020, 63(1):22-28.
[22] 雷妍圆, 章玉苹, 薛志洪, 等. 一株球孢白僵菌的分离鉴定及其对草地贪夜蛾的致病性[J]. 环境昆虫学报, 2020, 42(3):593-601.
[23] 蒲蛰龙, 李增智. 昆虫真菌学[M]. 合肥:安徽科学技术出版社, 1996.
[24] 邹东霞, 史国英, 廖旺姣, 等. 桉蝙蛾高毒力白僵菌菌株筛选及鉴定[J]. 中国森林病虫, 2016, 35(5):1-4.
[25] 黄鹏, 姚锦爱, 余德亿, 等. 虫生真菌BB-T02分离鉴定及其对两种蓟马的致病力[J]. 中国生物防治学报, 2020, 36(6):929-937.
[26] O'Donnell K, Sutton D A, Fothergill A, et al. Molecular phylogenetic diversity, multilocus haplotype nomenclature, and in vitro antifungal resistance within the Fusarium solani species complex[J]. Journal of Clinical Microbiology, 2008, 46(8):2477-2490.
[27] Mbofung G Y, Hong S G, Pryor B M. Phylogeny of Fusarium oxysporum f. sp. lactucae inferred from mitochondrial small subunit, elongation factor 1-alpha, and nuclear ribosomal intergenic spacer sequence data[J]. Phytopathology, 2007, 97(1):87-98.
[28] 吴益东, 沈晋良, 陈进, 等. 用点滴法和浸叶法监测棉铃虫抗药性的比较[J]. 植物保护, 1996, 22(5):3-6.
[29] 王联德, 尤民生, 黄建, 等. 虫生真菌多样性及其在害虫生物防治中的作用[J]. 江西农业大学学报, 2010, 32(5):920-927.
[30] 李宏科. 虫生真菌-镰刀菌的研究[J]. 湖南农业科学, 1989(4):36-37.
[31] 李宏科. 水稻害虫上三种病原镰刀菌的研究[J]. 植物保护学报, 1983, 10(4):217-224.
[32] 舒正义, 王中康, 尹勇. 四川地区嗜蚧镰刀菌的培养和侵染研究[J]. 西南农业大学学报, 1992, 14(3):244-246.
[33] 王辉, 杨志荣, 朱文, 等. 龟蜡蚧病原真菌的分离及应用研究[J]. 西南农业大学学报, 1999, 21(4):45-49.
[34] Claydon N, Grove J F, Pople M. Insecticidal secondary metabolic products from the entomogenous fungus Fusarium larvarum[J]. Journal of Invertebrate Pathology, 1979, 33(3):364-367.
[35] 苑森行, 杨玉景. 虫生真菌——木贼镰刀菌的初步研究[J]. 微生物学通报, 1986(1):13-14.
[36] 周与良, 邢来君. 一株从高粱蚜分离的镰刀菌的鉴定[J]. 昆虫学报, 1979, 22(2):217-218.
[37] 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.
[38] Ahmad F, Anwar W, Javed M A, et al. Infection mechanism of Aspergillus and Fusarium species against Bemisia tabaci[J]. Mycopath, 2019, 17(2):69-78.
[39] Chehri K. Molecular identification of entomopathogenic Fusarium species associated with Tribolium species in stored grains[J]. Journal of Invertebrate Pathology, 2017, 144:1-6.
[40] 高文彬, 田颖超, 董乾, 等. 异色瓢虫共生真菌木贼镰刀菌次生代谢产物研究[J]. 河南师范大学学报(自然科学版), 2019, 47(6):110-112.
[41] Sharma L, Marques G. Fusarium, an entomopathogen-a myth or reality?[J]. Pathogens, 2018, 7(4):93.
[42] de H. C. Maciel M, Amaral A C T D, Silva T D D, et al. Evaluation of mycotoxin production and phytopathogenicity of the entomopathogenic fungi Fusarium caatingaense and F. pernambucanum from Brazil[J]. Current Microbiology, 2021, 78(4):1218-1226.