应用荧光原位杂交方法检测中国沿海塔玛/链状亚历山大藻复合种(亚洲温带基因型)  

Application of fluorescence in situ hybridization (FISH) method to detect “tamarense/catenella species complex” (“Temperate Asian” ribotype) in Genus Alexandrium along Chinese coast

作　　者：于仁成[1] 唐祥海[1] 张清春[1] 陈洋[1] 王云峰[1] 颜天[1] 周名江[1] 

机构地区：[1]中国科学院海洋研究所海洋生态与环境科学重点实验室,青岛266071

出　　处：《环境科学学报》2006年第4期646-651,共6页Acta Scientiae Circumstantiae

基　　金：国家高技术研究发展计划(863)项目(No.2002AA639420);国家重点基础研究项目(No.2001CB409704);中国科学院海洋研究所知识创新项目(No.200223107)~~

摘　　要：近年来,有害赤潮于中国沿海频繁发生,对沿海居民身体健康、水产养殖和自然生态形成了潜在的威胁.由于部分有毒赤潮藻在很低的密度下就有可能导致严重的危害,但传统的采样和分析方法无法对这类有毒赤潮进行有效的检测,因而急需发展准确、高效的检测新方法.根据对中国沿海分离的塔玛/链状亚历山大藻(亚洲温带基因型)核糖体大亚基DNA(LSUrDNA)序列信息的分析,设计了两条特异性的荧光标记探针,并建立了针对中国沿海塔玛/链状亚历山大藻复合种(亚洲温带基因型)的荧光原位杂交检测方法.室内模拟实验显示两条探针都能够特异性地标记中国沿海塔玛/链状亚历山大藻(亚洲温带基因型),但标记效果有一定差异,探针SPEC-PROBE2标记效果远好于探针SPEC-PROBE1.经过标记的藻细胞可以通过荧光显微镜和流式细胞仪区分.中国沿海塔玛/链状亚历山大藻荧光原位杂交检测方法的建立将有助于提高海水样品中亚历山大藻监测的准确性和工作效率.The intensities and frequencies of harmful algal blooms （HABs） along the coast of China have been increasing dramatically in the past two decades, posing potent threats to the natural ecosystems, human health and aquaculture industry. Some toxic HAB species could lead to serious impacts even when the cell concentration in seawater is very low. Therefore, new techniques with high accuracy and efficiency for monitoring toxic species need to be developed since traditional monitoring methods could not fulfill the requirements. Here we report the development in fluorescence in situ hybridization （FISH） method to detect the ＂Alexandrium tamarense/catenella species complex＂ （ ＂Temperate Asian＂ ribotype） along Chinese coast, using two specific FITC-labeled probes targeted on Large Sub-Unit ribosomal RNA （ LSU rRNA）. The results showed that both of the probes could identify the toxic species specifically, but the labeling efficiency of probe SPEC-PROBE2 was much better than that of SPEC-PROBE1. The probe-labeled cells could be easily identified using fluorescence microscope or flow-cytometry. The application of the technique would improve the accuracy and efficiency in routine monitoring of Alexandrium species in China.

关 键 词：塔玛/链状亚历山大藻复合种 遗传探针 核糖体RNA 荧光原位杂交 

分 类 号：X835[环境科学与工程—环境工程] X171.5