Institutional Repository of Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences
| Development of taxonomic rRNA-targeted probes of two harmful algae: Prorocentrum minimum and Kareniamikimotoi by fluorescence in situ hybridization | |
| Chen Guofu1,2; Wang Quanfu1; Zhang Chunyun1; Zhang Baoyu3; Wang Guangce3; Lu Douding4; Xu Zhong1; Yan Peishen1; Wang, QF | |
| 2013-02-01 | |
| 发表期刊 | ACTA OCEANOLOGICA SINICA
 |
| ISSN | 0253-505X |
| 卷号 | 32期号:2页码:66-75 |
| 文章类型 | Article |
| 摘要 | Harmful algal blooms recently have been under the spotlight throughout the world, because of their negative impact on the marine environment, aquaculture, fisheries as well as public health. The development of methods for rapid and precise identification and quantification of causative species is essential for the warning andmonitoring of blooms, among which the techniques based on taxonomic probes are themost favored. In this study, two harmful algae, i.e., Prorocentrum minimum and Karenia mikimotoi were taken into consideration. The partial large subunit rDNA (D1-D2) of both species were firstly PCR-amplified, cloned and sequenced. The obtained sequences were then introduced to carry out alignment analysis for gene specific regions. Three respective candidate probes for each species were designed and used to screen the optimal probe by performing fluorescence in situ hybridization (FISH) tests. The results showed that the probes Pmin0443 and Kmik0602 displayed the best hybridization for P. minimum and K. mikimotoi, respectively. Both the specific (taxonomic) (Pmin0443 and Kmik0602) and the control probes (UniC0512 and UniR0499) were used for cross-reactivity tests with other microalgae in our laboratory. The probes Pmin0443 and Kmik0602 are specific and could be served as taxonomic probes introduced into the techniques targeting rRNA, such as FISH, sandwich hybridization, and DNA-microarray assay of P. minimum and K. mikimotoi in the future. Finally, FISH analyses with both probes were performed on the simulated field samples. The probes could hybridize exclusively with the target cells well, and no significant difference (p > 0.05) was observed in the cell densities of the samples determined by FISH and light microscopy (LM). All suggest that the probes are specific and could be introduced into FISH for the monitoring of both harmful algae.; Harmful algal blooms recently have been under the spotlight throughout the world, because of their negative impact on the marine environment, aquaculture, fisheries as well as public health. The development of methods for rapid and precise identification and quantification of causative species is essential for the warning andmonitoring of blooms, among which the techniques based on taxonomic probes are themost favored. In this study, two harmful algae, i.e., Prorocentrum minimum and Karenia mikimotoi were taken into consideration. The partial large subunit rDNA (D1-D2) of both species were firstly PCR-amplified, cloned and sequenced. The obtained sequences were then introduced to carry out alignment analysis for gene specific regions. Three respective candidate probes for each species were designed and used to screen the optimal probe by performing fluorescence in situ hybridization (FISH) tests. The results showed that the probes Pmin0443 and Kmik0602 displayed the best hybridization for P. minimum and K. mikimotoi, respectively. Both the specific (taxonomic) (Pmin0443 and Kmik0602) and the control probes (UniC0512 and UniR0499) were used for cross-reactivity tests with other microalgae in our laboratory. The probes Pmin0443 and Kmik0602 are specific and could be served as taxonomic probes introduced into the techniques targeting rRNA, such as FISH, sandwich hybridization, and DNA-microarray assay of P. minimum and K. mikimotoi in the future. Finally, FISH analyses with both probes were performed on the simulated field samples. The probes could hybridize exclusively with the target cells well, and no significant difference (p > 0.05) was observed in the cell densities of the samples determined by FISH and light microscopy (LM). All suggest that the probes are specific and could be introduced into FISH for the monitoring of both harmful algae. |
| 关键词 | Prorocentrum Minimum Karenia Mikimotoi Fluorescence In Situ Hybridization Taxonomic Probe |
| 学科领域 | Oceanography |
| DOI | 10.1007/s13131-013-0278-4 |
| URL | 查看原文 |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS研究方向 | Oceanography |
| WOS类目 | Oceanography |
| WOS记录号 | WOS:000314699900009 |
| WOS关键词 | WHOLE-CELL HYBRIDIZATION ; DINOFLAGELLATES ALEXANDRIUM-TAMARENSE ; SANDWICH HYBRIDIZATION ; OLIGONUCLEOTIDE PROBES ; RAPID IDENTIFICATION ; HETEROSIGMA-AKASHIWO ; COASTAL WATERS ; BACILLARIOPHYCEAE ; CATENELLA ; FIELD |
| WOS标题词 | Science & Technology ; Physical Sciences |
| 引用统计 | |
| 文献类型 | 期刊论文 |
| 条目标识符 | http://ir.qdio.ac.cn/handle/337002/16561 |
| 专题 | 实验海洋生物学重点实验室 |
| 通讯作者 | Wang, QF |
| 作者单位 | 1.Harbin Inst Technol Weihai, Weihai 264209, Peoples R China 2.State Ocen Adm, Inst Oceanog 1, Qingdao 266061, Peoples R China 3.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China 4.State Ocen Adm, Inst Oceanog 2, Hangzhou 310012, Peoples R China |
| 推荐引用方式 GB/T 7714 | Chen Guofu,Wang Quanfu,Zhang Chunyun,et al. Development of taxonomic rRNA-targeted probes of two harmful algae: Prorocentrum minimum and Kareniamikimotoi by fluorescence in situ hybridization[J]. ACTA OCEANOLOGICA SINICA,2013,32(2):66-75. |
| APA | Chen Guofu.,Wang Quanfu.,Zhang Chunyun.,Zhang Baoyu.,Wang Guangce.,...&Wang, QF.(2013).Development of taxonomic rRNA-targeted probes of two harmful algae: Prorocentrum minimum and Kareniamikimotoi by fluorescence in situ hybridization.ACTA OCEANOLOGICA SINICA,32(2),66-75. |
| MLA | Chen Guofu,et al."Development of taxonomic rRNA-targeted probes of two harmful algae: Prorocentrum minimum and Kareniamikimotoi by fluorescence in situ hybridization".ACTA OCEANOLOGICA SINICA 32.2(2013):66-75. |
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