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为了揭示不同蛋白酶抑制剂对绿僵菌致病力的影响,明确绿僵菌经消化道侵染东亚飞蝗的作用机制,采用分光光度法测定了经绿僵菌IPPM202及不同蛋白酶抑制剂处理后,不同时间东亚飞蝗中肠总蛋白酶、类胰蛋白酶、类胰凝乳蛋白酶、乙酰胆碱酯酶(Ach E)和谷胱甘肽硫转移酶(GSTs)活力的动态变化。结果表明,绿僵菌IPPM202处理后10 d内,东亚飞蝗中肠总蛋白酶活性呈现先上升再下降再上升的趋势,类胰蛋白酶活性呈先上升再下降趋势,类胰凝乳蛋白酶活性逐渐上升;但两种解毒酶的变化均为先上升后下降。加入两种抑制剂后,总体上抑制了东亚飞蝗中肠总蛋白酶和两种解毒酶的活性,降低了菌株IPPM202的侵染力,其中,甲苯磺酰基-L-氨基联苯氯甲基酮(tosyl-L-phenylalanine chloromethyl ketone,TPCK)作用较对脒苯基甲磺酰氯(4-amidino phenyl methane sulfonyl fluoride,APMSF)更加显著,使各种酶活力降低至与对照无差异水平。本研究对于揭示绿僵菌在中肠的致病机理及寄主对真菌侵染的免疫机制有一定的指导意义。
In order to reveal the effect of different protease inhibitors on the pathogenicity of Metarhizium anisopliae, the mechanism of Metarhizium anisopliae infecting Locusta migratoria manillensis by the digestive tract was clarified. After treated with IPPM202 and different protease inhibitors , Dynamic changes of total protease, tryptase, chymotrypsin, acetylcholinesterase (AchE) and glutathione S-transferase (GSTs) activity in midgut of Locusta migratoria manilensis at different times. The results showed that within 10 days after treatment with IPPM202, the total protease activity of midgut of the migratory locust showed upward trend, then downward trend and then upward trend. The tryptase activity increased first and then decreased, and the chymotrypsin activity gradually increased However, the changes of the two detoxification enzymes first increased and then decreased. The addition of two inhibitors inhibited the activity of total protease and two detoxification enzymes in the midgut of Locusta migratoria manilensis and reduced the infectivity of the strain IPPM202. Toluene sulfonyl-L-aminobiphenyl chloromethyl ketone The effect of tosyl-L-phenylalanine chloromethyl ketone (TPCK) was more significant than 4-amidino phenyl methane sulfonyl fluoride (APMSF), which reduced the activities of various enzymes to the same level as the control. This study has certain guiding significance for revealing the pathogenic mechanism of Metarhizium anisopliae in the midgut and the host immune mechanism to fungal infection.