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丁莉莉,罗忍,潘琦,王翠伦,于士将,丛林,刘浩强,李鸿筠,冉春.柑橘粉虱寄生性真菌渐狭蜡蚧菌TN002菌株培养条件的探究[J].中国南方果树,2017,46(2):
柑橘粉虱寄生性真菌渐狭蜡蚧菌TN002菌株培养条件的探究
The study on cultivating conditions of Lecanicillium attenuatum TN002 strain isolated from citrus whitefly
投稿时间:2016-11-30  修订日期:2016-12-15
DOI:
中文关键词:  渐狭蜡蚧菌  基本生物学特性  摇瓶培养
英文关键词:Lecanicillium attenuatum  biological characteristics  liquid cultivation
基金项目:国家科技支撑计划课题(2012BAD19B06);南方山地果园智能化管理技术与装备协同创新中心开放基金(编号:IX2016XCHJ01);中央高校基本业务费(编号:SDJK2015C085)
作者单位E-mail
丁莉莉 中国农业科学院柑桔研究所(西南大学柑桔研究所) dinglijiayoua@126.com 
罗忍 中国农业科学院柑桔研究所(西南大学柑桔研究所) koalox@163.com 
潘琦 中国农业科学院柑桔研究所(西南大学柑桔研究所) 17725015153@163.com 
王翠伦 中国农业科学院柑桔研究所(西南大学柑桔研究所) cuilunwang@163.com 
于士将 中国农业科学院柑桔研究所(西南大学柑桔研究所) yushijiang@sinocitrus.com 
丛林 中国农业科学院柑桔研究所(西南大学柑桔研究所) conglin@cric.cn 
刘浩强 中国农业科学院柑桔研究所(西南大学柑桔研究所) liuhaoqiang@cric.cn 
李鸿筠 中国农业科学院柑桔研究所(西南大学柑桔研究所) lihongjun@cric.cn 
冉春* 中国农业科学院柑桔研究所(西南大学柑桔研究所) ranchun@cric.cn 
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中文摘要:
      渐狭蜡蚧菌(Lecanicillium attenuatum)是一种地理分布和寄主范围广泛的重要昆虫病原真菌,具有潜在的生物防治应用前景。本研究以产孢量和菌落直径,菌丝干重为测定指标设计单因素实验,对从重庆地区柑桔粉虱上分离得到的渐狭蜡蚧菌TN002菌株的基本生物学特性和摇瓶培养条件进行了探究,以期为TN002菌株商业化生产提供理论依据。结果表明,渐狭蜡蚧菌TN002菌株在PDA培养基上10℃~35℃范围内均可生长,25℃条件下产孢量和菌落直径最大。不同pH对其生长产孢影响差异不大;pH=5.5时产孢量最大达到(6.52+-0.45)x10^8 个孢子/皿。有机氮更利于TN002菌株固体生长和产孢,其中酵母浸粉为氮源时菌落直径最大达(41.91+-0.46)mm,蛋白胨为氮源时产孢量最多,为(8.56+-0.4)x10^8 个孢子/皿。蔗糖和葡萄糖较利于TN002菌株固体生长和产孢。在葡萄糖酵母浸粉液体培养基培养下,产孢量能达到最大值为(8.08+-0.16)x10^8 个孢子/mL。摇瓶培养24 h~93 h的结果表明在60 h时TN002菌株产孢量和菌丝干重均达到最大值。对装量、转速和接种量进行正交设计实验,结果表明最优的处理组合为500 mL锥形瓶中装量100 mL,接种量为4%(孢子悬液浓度为:1x10^7 个孢子/mL),转速为190 r/min或220 r/min。该研究明确了渐狭蜡蚧菌TN002菌株生物学特性,筛选得到了摇瓶培养最佳条件,为进一步扩大培养提供了理论基础。
英文摘要:
      Lecanicillium attenuatum is an entomopathogenic fungus with a wide range of geographical distribution and host, has a high potential in the biological control of pests. In this work, the biological characteristics and the condition of liquid cultivating of the Lecanicillium attenuatum TN002 strain isolated from whitefly body in Chongqing were investigated, which provided theory basis for the commercial production of TN002 strain. The single factor experiments and orthogonal experiments were designed to measure the spore production, colony diameter and dry weight of mycelium. The result indicated that it was suitable for TN002 strain to grow and spore production at 10 to 35℃, while the best temperature was 25℃. The influence of the differential pH value on colonial growth and mycelial dry weight were not significant, and the maximal (6.52+-0.45)x10^8 spores per dish was found at pH 5.5. The organic nitrogen was benefits for the growth and yield of spores. The colony diameter cultured with yeast extract powder was the largest (41.91+-0.46 mm), while, the strain cultivated with peptone showed a maximal yield of conidial production (8.56+-0.4)x10^8 spores per dish. Sucrose and glucose were the best carbon sources for TN002 strain. The conidial production would reach the maximum when media prescription was selected as the glucose yeast extract, and the maximal conidial production was (8.08+-0.16)x10^8 spores.mL-1. In addition, the highest conidial production and mycelial dry weight were observed at 60 h after shaker cultivation for 24 h~93 h. The influence of volume of culture medium, initial conidial concentration and rotation speed on the spore production was studied utilizing orthogonal test. The optimal growing condition of spore production as the inoculum size was 4% (1x10^7 spores.mL-1 ) in 100 mL medium at 190 r.min-1 or 220 r.min-1. The biological characteristics of TN002 strain were ascertained and the optimum condition of liquid cultivating was obtained, which provided theory basis for the utilization of TN002 strain.
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