\r王洁华，毛虹禹，杨少辉\r
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AuthorsHTML:\r王洁华，毛虹禹，杨少辉\r
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AuthorsListE:\rWang Jiehua，Mao Hongyu，Yang Shaohui\r
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AuthorsHTMLE:\rWang Jiehua，Mao Hongyu，Yang Shaohui\r
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Unit:\r\r\r天津大学环境科学与工程学院，天津 300072\r
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Unit_EngLish:\rSchool of Environmental Science and Engineering，Tianjin University，Tianjin 300072，China\r
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Abstract_Chinese:\rKunitz 型胰蛋白酶抑制剂(Kunitz-type trypsin inhibitor，KTI)属于蛋白酶抑制剂(protease inhibitor，PI)家族中的丝氨酸蛋白酶抑制剂．本研究选择黑杨(Poplus nigra，Pn)中4 个可被机械创伤诱导的KTI 基因(PnKTIA6、PnKTIB1、PnKTIC1、PnKTID2)，在拟南芥中过表达并以转基因拟南芥叶片饲喂小菜蛾，观察其从幼虫到成蛾的整
个生长发育周期生理指标，从而检测4 种PnKTI 基因的抗虫效果．结果发现，4 种PnKTI 基因的抗虫效果依次为35,S：PnKTIA6＞35,S：PnKTID2＞35,S：PnKTIB1＞35,S：PnKTIC1，其中，过表达PnKTIA6 基因的拟南芥叶片饲喂的小菜蛾存活率最低，仅为9.8%,．此外，采用酶化学法测定分析了PnKTIs 基因编码产物的功能及效果．结果发现，PnKTIs 基因的编码产物对胰凝乳蛋白酶没有抑制作用，但对胰蛋白酶有很强的抑制作用，作用效果依次为35,S：PnKTIA6＞35,S：PnKTID2＞35,S：PnKTIB1＞35,S：PnKTIC1，与小菜蛾抗性测定结果一致．\r
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Abstract_English:\rThe Kunitz-type trypsin inhibitor(KTI) is a serine protease inhibitor that belongs to the protease inhibitor(PI)family. In this study，we selected four KTI gene (PnKTIA6，PnKTIB1，PnKTIC1，and PnKTID2) induced by mechanical trauma in Populus nigra，which were overexpressed in Arabidopsis thaliana that fed on Plutella xylostella to observe the physiological indicators of the entire growth and development cycle from larvae to adult moths and analyze their anti-insect effects. The results showed that the anti-insect effects of the four PnKTI genes were in the order 35,S：PnKTIA6＞35,S：PnKTID2＞35,S：PnKTIB1＞35,S：PnKTIC1. Among them，the survival rate of Plutella xylostella fed with Arabidopsis thaliana over-expressing PnKTIA6 was only 9.8%. We also applied enzymatic
chemistry to determine the function and effect of the PnKTI gene-encoding product，which showed that the PnKTI encoding product had no inhibitory effect on chymotrypsin，but had a strong inhibitory effect on trypsin. The effects were in the order 35,S：PnKTIA6＞35,S：PnKTID2＞35,S：PnKTIB1＞35,S：PnKTIC1，which were consistent with the result of Plutella xylostella resistance determination.\r
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Keyword_Chinese:PnKTI；拟南芥；小菜蛾；抗性；功能分析\r

Keywords_English:PnKTI；Arabidopsis thaliana；Plutella xylostella；resistance；functional analysis\r


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