短时高温暴露下东亚飞蝗雌虫血淋巴蛋白表达分析及质谱鉴定

Expression analysis and identification of proteins in hemolymph of Locusta migratoria manilensis Meyen(Orthoptera:Acrididae ) under short-term exposure to high temperature

为研究短时高温对东亚飞蝗Locusta migratoria manilensis Meyen血淋巴蛋白的影响,采用Bradford法、SDSPAGE电泳和质谱等方法,对东亚飞蝗雌虫血淋巴样品进行检测。结果表明:短时高温对血淋巴蛋白含量有显著影响(P<0.01),36℃-42℃范围内,随温度升高,血淋巴蛋白浓度亦升高,其中39℃、42℃处理组与对照组差异显著(P<0.01);短时高温对血淋巴蛋白种类存在一定影响,对照组雌虫血淋巴中存在11种蛋白,高温处理后,4种蛋白含量逐渐增加,6种蛋白含量没有明显变化,1种蛋白消失;经质谱检测,鉴定了5种蛋白,分别为载脂蛋白前体、酚氧化酶原、2个储存蛋白和19 kDa血淋巴蛋白,另外6条蛋白未被鉴定。推测载脂蛋白前体、酚氧化酶原、储存蛋白在东亚飞蝗应对高温胁迫过程中发挥重要作用。

In order to study the effect of short-term exposure to high temperature on proteins in hemolymph of Locusta migratoria manilensis Meyen,proteins in hemolymph of female adult locust were detected by Bradford assay,SDS-PAGE electrophoresis and MALDI-TOF-TOF/MS analysis.The results showed that short-term exposure to high temperature did a significant effect on concentration of protein in hemolymph(P<0.01).The concentration was increased with increasing temperature in the range of 36℃-42℃.There had significant increase on concentration between the treatment group(39℃,42℃)and the control(P<0.01).Short-term exposure to high temperature affected types of proteins in hemolymph.There had 11 types of proteins in hemolymph in the control.With increasing temperature,the concentration of 4 types of proteins were increased while 6 types of proteins showed no changed.Under different temperature treatments,1 type of proteins was disappeared.By MALDI-TOF-TOF/MS analysis,5 types of proteins were identified as apolipophorin precursor protein,pro-phenoloxidase 2,hexamerin-like protein 1,hexamerin-like protein 2 and basic 19 kDa hemolymph protein.The other 6 types of proteins were not identified.Apolipophorin precursor protein,pro-phenoloxidase,hexamerin protein may play pivotal roles in response to high temperature stress.