细胞凝集素探针对典型赤潮生物的定量检测  

Quantitative detection of harmful algea species using lectin probes

作　　者：黄辉[1] 武模戈[2] 雷红灵[3] 赖红艳[3] 侯建军[3] 

机构地区：[1]西南大学水产系,重庆荣昌402460 [2]濮阳职业技术学院生物系,河南濮阳457000 [3]湖北民族学院湖北省生物资源保护与利用重点实验室,湖北恩施445000

出　　处：《中国水产科学》2008年第2期279-292,共14页Journal of Fishery Sciences of China

基　　金：国家重点基础研究发展规划973项目(CEOHAB2001CB409704);厦门市创新科技基金项目(3502Z20041059);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)

摘　　要：采用14种异硫氰酸荧光素(FITC)标记的细胞凝集素探针对23株典型的有害赤潮生物进行了检测,结合流式细胞仪、荧光显微镜、荧光分光光度计对探针标记区分和识别目标藻的特异性和有效性进行定性和定量测定。结果表明,DBA可以把裸甲藻(Gymnodinium)GIXM01株与其他裸甲藻区分开;SBA、WGA和PNA探针能定性或定量地区分形态相似的裸甲藻GMDH01和TPXM01。PHA-E和SJA能分别将塔玛亚历山大藻(Alexandrium tamarense)ATDH04、ATMJ02株系与同种其他的藻株区分开;PNA也能从同种其他株系中识别塔玛亚历山大藻ATDH01,ATDH02,ATDH03株系;UEA I能从同种藻株中识别塔玛亚历山大藻ATCI01-JN、ATCI01藻株;RCA120可以区分产毒水平不同的亚历山大藻属(Alexandrium)的ASPGX01和ASPGX02。DBA、SBA、PNA和LCA探针能定性和定量区分微小原甲藻(Prorocentrum minimum)PMDH01和PMXM01;而PNA探针仅能定量区分原甲藻属(Prorocentrum)PDDH01与PMXM01。用WGA探针结合流式细胞仪、荧光显微镜、荧光分光光度等检测手段可以把探针标记的目标裸甲藻GSPXM01和TPXM01从米氏凯伦藻(Karenia mikimotoi)、东海原甲藻(P.donghaiense)和微小原甲藻(P.minimum)等组成的混合样品中区分开。Lectins are carbohydrate-binding proteins of nonimmune origin that agglutinate cells or precipitate glycoconjugates, and they have been found in a wide range of organisms from bacteria to higher vertebrates. They are non-enzymatic secretory proteins or glycoproteins that play important physiological roles as recognition molecules within cell, between cells, or between organisms. Because of the ability of lectins to discriminate specific carbohydrate structures and the molecules＇ biological activities, lectins are useful as probes for carbohydrate structures studies in many areas of biological research. Many researches have been done using lectins to detect and differentiate harmful algae. However, there has been limited work on quantification of bound lectins, and the lectin probes used are not specific enough to detect HAB species whose taxonomies are still unclear. There has been little work on utilization of FITC-conjugated lectins for identifying and differentiating harmful microalgae collected from coastal waters in China. In the present study, 14 lectin probes were used to assess their specificity （both quality and quantity） on Chinese harmful algae using different detection methods, such as flow cytometry （FCM）, epifluorescence microscopy （EFM） and spectrofluorometer （SFM）. The purpose of the study was to identify lectins that can distinguish morphologically similar, toxic and nontoxic algae in coastal waters in China. Results showed that of the tested lectin probes, except GMDH01 strain to each of the tested Gymnodinium species bound to most probe PHA-E, SBA, WGA and GSL Ⅰ, TPXM01 strain to probe PHA-E and GSL Ⅰ, CCMP1740 strain to probe PHA-E. GIXM01 could be be differentiated by DBA probe from other Gymnodinium species, and SBA, WGA could discriminate morphologically similar GMDH01 and TPXM01 from each other. The efficiency of GSL Ⅰ and PHA-E probe was not strong enough to distinguish different Gymnodinium species. Although GMDH01 and TPXM01 were similar in morphology and both co

关 键 词：细胞凝集素探针 赤潮种类 检测 流式细胞仪 荧光显微镜 荧光分光光度计 

分 类 号：X171[环境科学与工程—环境科学]