弹尾虫单克隆抗体的制备及其在捕食研究中的应用  

Development of a monoclonal antibody to Collembola and its application on predation evaluation

作　　者：赵伟春[1] 王光华[1] 程家安[1] 陈正贤[2] 

机构地区：[1]浙江大学昆虫科学研究所 [2]浙江大学生物技术研究所,杭州310029

出　　处：《生态学报》2007年第9期3694-3700,共7页Acta Ecologica Sinica

基　　金：国家"973"资助项目(G2000016210);国家自然科学基金资助项目(30370934)~~

摘　　要：应用杂交瘤技术制备了针对弹尾虫的单克隆抗体2F10。该抗体的效价为1.024×108,只与灰橄榄长角跳虫、球角跳虫和钩圆跳虫等弹尾虫发生强烈反应而不与稻田常见的其它昆虫和蜘蛛发生交叉反应,具有高度特异性。建立了2F10、HRP-2F10和蜘蛛样品分别稀释4000倍(34.193ng/L)、1500倍(2.4624ng/L)和50倍(50ml/individual)的抗体夹心ELISA检测系统用于检测稻田常见蜘蛛对弹尾虫的捕食作用。其检测灵敏度为1/2头灰橄榄长角跳虫(4.49μg),拟环纹豹蛛捕食1头灰橄榄长角跳虫成虫后,在25℃下猎物的可测定时间为4.5h。应用该检测系统研究了不同稻区常见蜘蛛对2F10的阳性反应率。其中狡蛛、拟环纹豹蛛、纵条蝇狮和纵条蝇虎的阳性反应率显著高于食虫瘤胸蛛、八斑球腹蛛和锥腹肖蛸。Collembola is a common detritivore in many cropping system. In order to understand the impact of Collembola on predators and the prey shifting behaviour in rice ecosystem, research was carried out to develop a monoclonal antibody （McAb） for major species of Collembola for identification. This paper reports the characterization of the McAb and evaluation to detecting the Collembola diet of spiders. A McAb for Collembola, namely 2F10, was developed using hybridoma technique. It had high absorption values even it was diluted over 1. 024 × 10^8 times. It is specific to the proteins of Entomobrya griseoclivata （ Packcord）, Hypogastra rnatura and BourletieUa christianseni （Banks） , and has no-reaction with other species in subtropical rice ecosystem. A double antibody sandwich method of enzyme-linked immunosorbent assay （ELISA） was established to detect the Collembola antigens in the guts of the spiders. In this detecting system, 2F10 diluted 4000 times （34. 193 ng/L）, enzymelinked antibody HRP-2F10 diluted 1500 times （2. 463 ng/L） and samples diluted 50 times （50ml/individual） were used. About 4.49 μg prey antigens （1/2 a Collembola individual） remaining in a predator gut could be detected under this system. It could recognize E. griseoclivata proteins after one aduh was ingested by Pardosa pseudoannulata for about 4.5 hours at 25 ℃. Samples of major spider species collected from rice paddies in Hangzhou, Fuyang and Anji were studied using this system. The result showed that Dolomedes sp. , Paradosa pseudoannulata, Marpissa magister and Plexippus setipesp preyed significantly on more Collembola than Ummeliata insecticeps, Coleosoma octomaculatum and Tetragnatha sp.

关 键 词：弹尾虫 蜘蛛 单克隆抗体 酶联免疫吸附试验 捕食作用 

分 类 号：S186[农业科学—农业基础科学]