楚雄腮扁叶蜂虫生真菌粉红粘帚霉的鉴定及其生物学特性 |
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引用本文:芦俊佳,夏举飞,庄飞,徐荣,曾廷潇,李永和.楚雄腮扁叶蜂虫生真菌粉红粘帚霉的鉴定及其生物学特性.植物保护学报,2018,45(3):622-631 |
DOI:10.13802/j.cnki.zwbhxb.2018.2018007 |
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中文摘要:为明确楚雄腮扁叶蜂Cephalcia chuxiongica病原真菌的种类,以组织分离法对自然罹病死亡的楚雄腮扁叶蜂进行虫生真菌的分离培养,根据形态学特征和rDNA ITS序列分析对所得虫生真菌进行鉴定,并对其中的高毒力菌株进行生物学特性测定。结果表明:共分离得到4属25株菌株,其中以粉红粘帚霉Clonostachys rosea出现频率最高,共分离到15株,均与GenBank中粉红粘帚霉相关菌株亲缘关系最近,ITS序列同源性达100%,属优势种。按照柯赫法则证实4个属的虫生真菌对楚雄腮扁叶蜂均有致病力,其中粉红粘帚霉致病力最强,其高毒力菌株SWFUYHL 02-01对楚雄腮扁叶蜂幼虫致死中时为10 h;蛋白胨马铃薯葡萄糖琼脂培养基最适宜该菌株生长和产孢,其次是马铃薯葡萄糖琼脂培养基;该菌的最适生长温度为25℃,光照对其菌丝生长无显著影响;在以不同种类糖为碳源的培养基上该菌株均能生长,以蔗糖为碳源的培养基上产孢量最高,显著高于以其它糖为碳源时;适宜其生长的氮源是酵母浸出粉,适宜产孢的氮源是硝酸铵和尿素,以硫酸铵和酵母浸出粉为氮源时不产孢;适宜菌丝生长和产孢的无机盐分别是MnSO4和MgSO4,添加CaCl2的培养基上未见产孢;光照对粉红粘帚霉孢子萌发无显著影响。粉红粘帚霉菌株在采样点的出现频率与其生物学特性完全吻合。 |
中文关键词:楚雄腮扁叶蜂 虫生真菌 粉红粘帚霉 鉴定 生物学特性 |
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Identification and biological characterization of mycoparasitic fungus Clonostachys rosea isolated from sawfly Cephalcia chuxiongica |
Author Name | Affiliation | E-mail | Lu Junjia | College of Biodiversity Conservation and Utilisation, Southwest Forestry University, Kunming 650224, Yunnan Province, China | | Xia Jufei | Yunnan Dianzhong New District, Kunming 650000, Yunnan Province, China | | Zhuang Fei | College of Life Sciences, Southwest Forestry University, Kunming 650224, Yunnan Province, China | | Xu Rong | College of Biodiversity Conservation and Utilisation, Southwest Forestry University, Kunming 650224, Yunnan Province, China | | Zeng Tingxiao | College of Biodiversity Conservation and Utilisation, Southwest Forestry University, Kunming 650224, Yunnan Province, China | | Li Yonghe | College of Biodiversity Conservation and Utilisation, Southwest Forestry University, Kunming 650224, Yunnan Province, China | swfclyh@126.com |
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Abstract:To clarify the entomopathogenic fungal species of sawfly Cephalcia chuxiongica, the isolation and culture of the fungus on morbid Ce. chuxiongica were carried out on the basis of tissue separation. The isolated fungus was identified based on morphological and molecular evidences, and the biological characters of the highly pathogenic strain were determined. The results showed that 25 strains belonging to four genera were obtained, and the highest frequency was Clonostachys rosea, which contained 15 strains and was closely related to Cl. rosea registered in GenBank. Their maximum similarity were 100%; therefore, Cl. rosea was the dominant species. According to the Koch's postulate, the pathogenicity of Cl. rosea to Ce. chuxionggica was confirmed. Cl. rosea had the strongest pathogenicity among four genera, and the highest pathogenic strain was SWFUYHL 02-01, of which the LT50 was 10 h against the larvae of Ce. chuxiongica. The optimum solid medium for the growth and spore production of SWFUYHL 02-01 was PPDA, followed by PDA. The optimum temperature for growth was 25℃, and the effect of light was not obvious on the mycelial growth. This strain could grow on the medium with different sugars as carbon sources, and the optimum medium for conidial production was sucrose, significantly better than other sugars. The best nitrogenous source for mycelium growth was the yeast extract powder. NH4NO3 and urea were the best nitrogenous sources for conidial production, and there were no spores in the medium with (NH4)2SO4 and yeast extract powder for nitrogenous sources. The best inorganic salt for mycelial growth and conidial production were MnSO4 and MgSO4, respectively, and there were no spore production in the medium with CaCl2 added. The effect of light was not obvious on the spore germination of Cl. rosea. The frequency of Cl. rosea at the sampling sites was consistent with its biological characteristics. |
keywords:Cephalcia chuxiongica entomopathogenic fungus Clonostachys rosea identification biological characteristics |
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