Rapid detection of natural cells of Alexandrium tamarense and A-catenella (Dinophyceae) by fluorescence in situ hybridization
Rapid detection of natural cells of Alexandrium tamarense and A-catenella (Dinophyceae) by fluorescence in situ hybridization
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DOI：
10.1016/j.hal.2004.04.002
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发表时间：
2005-02-01
期刊：
影响因子：
6.6
通讯作者：
Sako, Y
中科院分区：
文献类型：
--
作者：
Hosoi-Tanabe, S;Sako, Y
根据常规监测的形态特征，对引起麻痹性贝类中毒(PSP)的海洋有毒甲藻Alexandrium Tamarense(LeBor)Balech和A.catenella(Whedon And Kofoid)Taylor进行了鉴定。然而，由于形态上的相似性，快速而简单的鉴定往往是困难的。利用核糖体RNA(RRNA)靶向探针的荧光原位杂交(FISH)已被研究为一种容易识别和计数有害藻华(HAB)物种的方法。然而，它在监测赤潮物种自然种群方面的应用是有限的。在此，我们应用FISH方法对采自日本近海的天然浮游生物群落中的塔玛螺旋藻和链状螺旋藻进行了鉴定和计数。根据大亚基核糖体RNA基因(28S RDNA)的D2区，建立了塔玛氏不动杆菌特异性(Atm1)和链球菌特异性(Act1)探针。利用这两种探针，在满足以下三个条件的情况下，可以很容易地鉴定出野外样品中的塔玛依虫或链球菌的天然细胞。首先，细胞应该通过过滤而不是离心法进行浓缩，以避免细胞的损失。其次，应尽量减少自发荧光；丙酮是一种有效的脱色剂。第三，样品应保存在-20℃或-80℃，以便长期保存。结果表明，FISH是一种快速鉴定有毒亚历山大藻的有用工具。SPP.并可方便对大量天然样品进行分析。(C)2004爱思唯尔B.V.保留所有权利。
The marine toxic dinollagellates Alexandrium tamarense (Lebor) Balech and A. catenella (Whedon and Kofoid) Taylor that cause paralytic shellfish poisoning (PSP) are identified on the basis of morphological features in routine monitoring. Rapid and simple identification is, however, often difficult because of the morphological similarity. Fluorescent in situ hybridization (FISH) using ribosomal RNA (rRNA)-targeted probes has been studied as a method of easily identifying and enumerating species responsible for harmful algal blooms (HABs). Its application to monitoring natural populations of HAB species, however, is limited. Here, we applied the FISH method to identify and enumerate cells of A. tamarense and A. catenella in natural plankton assemblages collected from Japanese coastal waters. A. tamarense-specific (Atm1) and A. catenella-specific (Act1) probes were established based on the D2 region of the large-subunit ribosomal RNA gene (28S rDNA). With these two probes, natural cells of A. tamarense or A. catenella in field samples could easily be identified when the following three conditions were met. First, cells should be concentrated by filtration, not centrifugation, in order to avoid the loss of cells. Second, autofluorescence should be minimized; acetone was an effective decolorization reagent. Third, samples should be stored at -20 or -80 degreesC for long-term preservation. The results indicate that FISH is a useful tool for the rapid identification of toxic Alexandrium. spp. and can facilitate the analysis of numerous natural samples. (C) 2004 Elsevier B.V. All rights reserved.