用细胞周期分析结合免疫印迹方法检测微型浮游植物细胞增长率的研究  

Detection of In Situ Growth Rate of Phytoplankton Based on Cell-cycle Analysis and Western Blotting

作　　者：赖红艳[1] 武模戈[2] 侯建军[1] 刘绍平[3] 

机构地区：[1]湖北省生物资源保护与利用重点实验室(湖北民族学院) [2]濮阳职业技术学院生物系,河南濮阳457000 [3]农业部淡水鱼类种质资源与生物技术重点开放实验室(中国水产科学研究院长江水产研究所),湖北荆州434000

出　　处：《湖北民族学院学报（自然科学版）》2008年第1期92-96,103,共6页Journal of Hubei Minzu University(Natural Science Edition)

基　　金：中国博士后基金项目(20060400854);农业部淡水鱼类种质资源与生物技术重点实验室开放课题(LFB20070611);教育部博士点基金项目(20070504076);湖北省生物资源保护与利用重点实验室开放课题资助项目

摘　　要：采用基于流式细胞仪的细胞周期分析和免疫印迹方法来研究浮游植物的细胞增长率.检测和分析同步化后的有害浮游植物Takayama pulchellum,可知T.pulchellum的S期大约出现在10h左右.免疫印迹检测的结果显示抗微小原甲藻(Prorocentrum minimum)和抗Pfiesteria piscicida的PCNA抗体与T.pulchellum无明显的阳性杂交信号出现,表明这2种PCNA抗体不能作为检测T.pulchellum PCNA表达的抗体.Rubisco抗体免疫印迹反应的信号强烈,但与细胞生长状态的相关关系不够明显,其特异性不高使其不能作为理想的指示细胞原位生长率的指标.Estimating harmful algal bloom species＇ growth rates can be difficult to apply in the field, we have been exploring the feasibility of using proliferating cell nuclear antigen and Rubisco as the basis for making such estimates. In situ growth rate of phytoplankton was studied by western blotting and cell - cycle analysis based on flow cytometry. Cell - cycle progression of Takayam pulchellum at different times after synchronization was detected and analyzed and the results showed that S phase of T. pulchellum cell -cycle was at 10 h after synchronization. The result from western blotting exhibited that there were no positive antigenic reactions of Prorocentrum minimum and Pfiesteria piscicida -specific PCNA antibodies with T. pulchellum respectively, which suggested that the two antibodies could not be an immunofluores- cence assay to detect T. pulchellum. The detectable signal from western blotting of Rubisco antibody of P. minimum and P. piscicida was strong reacted with other algae tested, including T. pulchellum, but there was no obvious relationship between the detectable signal and its cell growth rate, and the results suggested that Rubisco was not too specific to be a perfect indicator of in situ growth rate of phytoplankton.

关 键 词：浮游植物 细胞周期分析 细胞增值核抗原 原位增长率 免疫印迹 

分 类 号：Q145[生物学—生态学]