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Development of a fluorescence in situ hybridization (FISH) method for rapid detection of Ulva prolifera
Zhang, Qing-Chun1; Liu, Qing1,2; Kang, Zhen-Jun1,2; Yu, Ren-Cheng1; Yan, Tian1; Zhou, Ming-Jiang1
2015-09-20
发表期刊ESTUARINE COASTAL AND SHELF SCIENCE
卷号163期号:2015页码:103-111
文章类型Article
摘要Large-scale green tides have occurred consecutively since 2007 in the Yellow Sea (YS), China. The dominant causative species of the green tides has been identified as Ulva prolifera. The origin of green tides in the YS has been traced back to the Subei Shoal based on the results of remote-sensing, numerical simulations and field investigations. However, it is difficult to study the early development of green tides in the Subei Shoal because of the mixture of multiple green algae and the morphological diversity of U. prolifera when under variable environmental conditions. In this study, a rapid and accurate fluorescence in situ hybridization (FISH) method was developed to detect U. prolifera from the community of green algae targeting the 5S rDNA spacer region of U. prolifera. Two specific probes, 5S-1 and 5S-2, were designed based on the sequences of the 5S rDNA spacer regions of U. prolifera, Ulva linza and Ulva flexuosa. Specificity of the FISH method was tested using the six species of green algae commonly occurring in the Subei Shoal, including U. prolifera, U. linza, U. flexuosa, Ulva compressa, Ulva pertusa and Blidingia sp. The results showed that only U. prolifera could be labeled with both probes. Probe 5S-1, which showed a much higher labeling efficiency on U. prolifera, was ultimately selected as the probe for the FISH detection. The sample preparation method was optimized, particularly for the mature green algae, by the addition of cellulase and proteinase K in the pre-hybridization solution. Labeling efficiency with the probe 5S-1 reached 96% on average under the optimized conditions. The successful development of the FISH method has been applied to qualitative and quantitative analysis of field samples collected from the YS, and the results indicate a potential use in future green algae studies. (C) 2014 Elsevier Ltd. All rights reserved.
关键词Fluorescence In Situ Hybridization (Fish) Ulva Prolifera Green Tide Yellow Sea
DOI10.1016/j.ecss.2014.09.004
收录类别SCI
语种英语
WOS记录号WOS:000362604700017
引用统计
被引频次:23[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.qdio.ac.cn/handle/337002/52135
专题海洋生态与环境科学重点实验室
作者单位1.Chinese Acad Sci, Inst Oceanol, Key Lab Marine Ecol & Environm Sci, Qingdao 266071, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
第一作者单位中国科学院海洋研究所
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Zhang, Qing-Chun,Liu, Qing,Kang, Zhen-Jun,et al. Development of a fluorescence in situ hybridization (FISH) method for rapid detection of Ulva prolifera[J]. ESTUARINE COASTAL AND SHELF SCIENCE,2015,163(2015):103-111.
APA Zhang, Qing-Chun,Liu, Qing,Kang, Zhen-Jun,Yu, Ren-Cheng,Yan, Tian,&Zhou, Ming-Jiang.(2015).Development of a fluorescence in situ hybridization (FISH) method for rapid detection of Ulva prolifera.ESTUARINE COASTAL AND SHELF SCIENCE,163(2015),103-111.
MLA Zhang, Qing-Chun,et al."Development of a fluorescence in situ hybridization (FISH) method for rapid detection of Ulva prolifera".ESTUARINE COASTAL AND SHELF SCIENCE 163.2015(2015):103-111.
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