不同分子探针对赤潮样品检测效果的比较研究  

Comparison of Efficiency for Different Molecular Probes Applied to Detect HAB Species in Natural Samples

作　　者：侯建军[1,2,3] 赖红艳[1] 武模戈[4] 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 [2]华中农业大学水产学院,湖北武汉430070 [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43400 [4]濮阳职业技术学院生物系

出　　处：《湖北民族学院学报（自然科学版）》2008年第3期249-255,共7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)

摘　　要：采用荧光标记的DNA和lectin探针对来自赤潮现场样品中的目标赤潮原因种进行了检测.结果表明这些探针能从比较复杂的现场样品背景中把目标赤潮生物识别出来.从检测效果上看,各种DNA探针对塔玛亚历山大藻、微小原甲藻和T.pulchellum标记识别的荧光信号强度相对于lectin探针要弱.Lectin探针与明视野下的显微镜计数结果呈现出很好的相关性,但lectin探针对目标藻的绝对检出数量比明视野下的计数结果要低.lectin探针(ConA)和DNA探针(PM18S02、PM28S02)对现场样品中微小原甲藻的检测计数结果也有较好的相关性,表明在这些检测条件下,不同的探针对特定目标藻的检测结果具有可比性、相关性和可操作性.Harmful Algal Blooms （HABs） are increasing globally. Rapid and unequivocal identification of toxic species has become a focal point of recent toxic algal research. Some new technologies such as molecular probes have been developed to differentiate non -toxic from toxic algae and to monitor the development of algal blooms in coastal waters. Probes of FITC - conjugated lectins and oligonueleotide （DNA） were applied to detect harmful algal bloom- forming species from natural samples in this study. The results showed that both lectin and DNA probes could identify these algal target cells from other negative algal cells and complicated background of natural samples. The detecting efficiency for Prorocentrum minimum, Alexandrium tamarense and Takayama pulcheilum by DNA and lectin probes was that the fluorescence intensity of algal ceils labeled by DNA was weaker than that by lectin probes. The correlation of cell counts for lectin probes in natural sample with those made by enumeration under the light microscope （LM） was good （R^2 =0.9778, WGA to LM for T. pulchellum;R^2 =0.9948, DBA to LM for A. tamatense; R^2 =0. 9980, ConA to LM for P. minimum）, but absolute cell numbers counted by lectin probes were less than that by light microscope. Cell counts of P. minimum enumerated by ConA probe and DNA probes （PMISSO2,PM28S02） respectively in different natural samples, the correlation was alse good （R^2 = 0.615, n = 28, P 〈 0. 05 ）. All results suggested that detection efficiency of different probes, such as DNA and lectin for identifying given HAB species were comparable, correlative and exercisable under the same detecting condition and method, and these probes and protocols might become useful tools to monitor in situ distribution of bloom - forming taxa, such as P. minimum, A. tamarense and T. pulchellum in natural water.

关 键 词：赤潮生物 DNA探针 细胞凝集素探针 原位荧光杂交 现场样品 

分 类 号：Q178.53[生物学—水生生物学] Q179.1[生物学—普通生物学]