海芋凝集素的纯化及其对4种昆虫细胞的毒杀作用  

Purification of a Lectin from Alocasia macrorrhiza and its Toxic Effect on Four Cultured Insect Cell Line

作　　者：潘科[1] 黄炳球[1] 侯学文[2] 

机构地区：[1]华南农业大学资源环境学院,广东广州510642 [2]华南农业大学生命科学学院,广东广州510642

出　　处：《安徽农业科学》2007年第18期5484-5485,5495,共3页Journal of Anhui Agricultural Sciences

基　　金：广东省自然科学基金项目(000643)

摘　　要：通过离体昆虫细胞培养研究海芋凝集素（Alocasia macrorrhiza lectin,AML）的毒杀作用。采用MTT法研究了提取自海芋叶0-30%、叶30%-80%、块茎0-30%、块茎30%-80%盐析段海芋凝集素粗品对4种昆虫细胞[草地贪夜蛾Spodoptera frugiperda Smith细胞（简称Sf-9细胞）、甜菜夜蛾Spodoptera exigua Hübner细胞（简称Se 301细胞）、粉纹夜蛾Trichoplusia ni Hübner细胞（简称Hi5细胞）和美洲棉铃虫Heliothis zea Boddie细胞（简称Hz细胞）]的毒杀作用,结果表明,海芋叶、块茎上述盐析段的粗提物均对4种昆虫细胞有明显的毒杀作用,其中以块茎30%-80%的样品毒杀活力最高：在2%浓度处理24 h条件下,上述4种细胞的相对死亡率分别为91.40%、91.62%、97.13%和82.30%。用Q Sepharose Fast Flow柱（2.6 cm×30 cm）对海芋块茎30%-80%的盐析物进行分离纯化,用最大活性峰样品对Sf-9细胞的毒杀作用进行研究,得到处理24 h的毒力回归方程,LC50为0.79%。The toxic effect of Alocasia macrorrhiza lectin （AML） on in vitro cultured insect cells was studied in this report. Four types of crude lectin were extracted and their toxic effects were tested with four insect cell lines which were Sf-9 from Spodoptera frugiperda Smith,Se301 from Spodoptera exigua Habner, Hi5 from Trichoplusia ni Htthner, and Hz from Heliothis zea Boddie, respectively. With MTT method, the obvious toxic effect of AML on four tested insect cell lines was found, with the 30% -80% tuber extracts showed the strongest effect on above cells,and the relative cell death rotes were 91.40%, 91.62% ,97.13% and 82.30% respectively,under 2% crude extracts concentration and 24 h treatment.This extract was further purified by Q Sepharose Fast Flow （2.6 cm ×30 cm）, and the highest activity peak was tested by Sf-9 cell. The toxicity regression equation under 24 h treatment was obtained,with the LC50 value of 0.79%.

关 键 词：海芋凝集素 昆虫细胞培养 MTT法 

分 类 号：Q946[生物学—植物学]