中国农业科学 ›› 2017, Vol. 50 ›› Issue (3): 591-598.doi: 10.3864/j.issn.0578-1752.2017.03.018

• 研究简报 • 上一篇    

球孢白僵菌对葱蝇成虫血淋巴蛋白质及游离氨基酸的影响

张慧1,吴圣勇1,王晓青2,雷仲仁1

 
  

  1. 1中国农业科学院植物保护研究所植物病虫害生物学国家重点实验室,北京 100193;2北京市植物保护站,北京 100029
  • 收稿日期:2016-06-13 出版日期:2017-02-01 发布日期:2017-02-01
  • 通讯作者: 雷仲仁,Tel:010-62815930;E-mail:leizhr@sina.com
  • 作者简介:张慧,E-mail:huizhang0926@sina.com
  • 基金资助:
    国家大宗蔬菜产业技术体系(CARS-25-B-07)、韭菜安全生产关键技术研究及科技示范基地建设(Z151100001215009)

Changes in the Contents of Proteins and Free Amino Acid in haemolymph of Delia antique Adult Infected by Beauveria bassiana
 

ZHANG Hui1, WU ShengYong1, WANG XiaoQing2, LEI ZhongRen1   

  1. 1State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193; 2Plant Protection Station of Beijing, Beijing 100029
  • Received:2016-06-13 Online:2017-02-01 Published:2017-02-01

摘要: 【目的】球孢白僵菌(Beauveria bassiana)是一种重要的昆虫病原真菌,在毁灭性害虫葱蝇(Delia antiqua)的防治上具有重大潜力。论文旨在明确球孢白僵菌侵染葱蝇成虫后,血淋巴中蛋白质和游离氨基酸的变化及其致病的关系,揭示球孢白僵菌对葱蝇的生理调控机制。【方法】基于球孢白僵菌对葱蝇成虫的室内生物测定的研究基础,使用菌株GZGY-1-3,采用点滴法,用浓度为1×108个孢子/mL的孢子悬浮液处理葱蝇初羽化成虫,采用BCA法及高效液相色谱——串联质谱法(HPLC-MS/MS)测定球孢白僵菌侵染不同时间(12、24、36、48、60和72 h)后,侵染组与对照组葱蝇成虫血淋巴中蛋白质总量及20种游离氨基酸含量;分析不同处理时间蛋白质和游离氨基酸总量和重要氨基酸的含量变化。【结果】球孢白僵菌侵染12—36 h,侵染组与对照组血淋巴蛋白浓度变化趋势相近,均呈先下降后小幅度上升趋势。但侵染组血淋巴中蛋白含量高于对照组,侵染24 h后,侵染组血淋巴内蛋白质是对照组的1.20倍;侵染48 h后,侵染组中蛋白含量下降到最低,仅为对照组的0.83倍。侵染60—72 h,血淋巴蛋白含量升高。葱蝇成虫血淋巴中共检测出20种游离氨基酸。侵染12—36 h,葱蝇成虫血淋巴中游离氨基酸总量下降;侵染36 h,游离氨基酸总量下降到最低,仅为对照组的0.63倍;侵染48—60 h,游离氨基酸总量持续上升;侵染60 h后,游离氨基酸总量上升到最高,为对照组的2.01倍;感染72 h,游离氨基酸总量下降。侵染后,血淋巴内游离氨基酸种类没有变化,各种游离氨基酸的含量波动差异较大。球孢白僵菌对血淋巴重要氨基酸的含量变化有显著影响。【结论】球孢白僵菌的侵染引起葱蝇成虫血淋巴中蛋白质和游离氨基酸的一系列变化,严重阻碍了葱蝇成虫的正常生理活动,在葱蝇生物防治中具有应用潜力。

关键词: 葱蝇, 球孢白僵菌, 血淋巴, 蛋白质, 游离氨基酸

Abstract: 【Objective】Beauveria bassiana is an important biological agent for control of Delia antiqua. The objective of this study is to assess the changes of proteins and free amino acid in haemolymph of D. antiqua adult infected by B. bassiana at different times, understand the relationship between the contents of proteins and free amino acids and the infection of B. bassiana.【Method】Based on the result of dose-response bioassay against D. antiqua adults infected by B. bassiana,individual newly enclosed D. antiqua adults were inoculated with 5 µL 1×108 conidia/mL conidial suspension of the GZGY-1-3 strain by micro-drop method. Then the contents of proteins and free amino acid of at different times (12, 24, 36, 48, 60 and 72 h) were measured in infected and uninfected D. antiqua adult using BCA and HPLC-MS/MS. Finally, the relationship between the infection and the contents of the key free amino acid was analyzed. 【Result】The content of proteins declined first and then increased at 12-36 h. The treatment group and control group showed similar changes. However the contents of proteins of the treatment group were higher than the control group. The content of proteins increased to the maximum in 24 h after inoculation, was 1.20 times greater than the control group. The content of proteins of the treatment group declined to the minimum at 48 h, was only 0.83 lower times than the control group, and increased at 60-72 h. The results of amino acids analysis showed that there were no different of free amino acid between the treatment and control groups of D. antiqua adult. The content of free amino acids decreased in 12-36 h, increased in 48-60 h and decreased in 72 h again after inoculation. The content of free amino acids declined to the minimum in 36 h after inoculation, was 0.63 times lower than the control group and increased to the maximum in 60 h after inoculation, was 2.01 times greater than the control group. After infection, there was no change in the type of free amino acids in the hamolymph, free amino acid content of various fluctuations was quite different. The infection of B. bassiana had significant influence on the contents of important amino acids in the haemolymph. 【Conclusion】 The B. bassiana invaded into the insect body affected the balance of the proteins and amino acids in the haemolymph. It seriously hampered the normal physiological activity of the D. antiqua adult. The results of the experiment indicated that B. bassiana strains couldeffectively control D. antiqua adult.

Key words: Delia antique, Beauveria bassiana, haemolymph, protein, free amino acid