naa a22 4500 510754708 CHVBK 20180411083118.0 cr unu---uuuuu 180411e20130201xx s 000 0 eng 10.1007/s13131-013-0278-4 doi (NATIONALLICENCE)springer-10.1007/s13131-013-0278-4 Development of taxonomic rRNA-targeted probes of two harmful algae: Prorocentrum minimum and Kareniamikimotoi by fluorescence in situ hybridization [Elektronische Daten] [Guofu Chen, Quanfu Wang, Chunyun Zhang, Baoyu Zhang, Guangce Wang, Douding Lu, Zhong Xu, Peishen Yan] 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. The Chinese Society of Oceanography and Springer-Verlag Berlin Heidelberg, 2013 Prorocentrum minimum nationallicence Kareniamikimotoi nationallicence fluorescence in situ hybridization nationallicence taxonomic probe nationallicence Chen Guofu Harbin Institute of Technology (Weihai), 264209, Weihai, China aut Wang Quanfu Harbin Institute of Technology (Weihai), 264209, Weihai, China aut Zhang Chunyun Harbin Institute of Technology (Weihai), 264209, Weihai, China aut Zhang Baoyu Institute of Oceanology, Chinese Academy of Sciences, 266071, Qingdao, China aut Wang Guangce Institute of Oceanology, Chinese Academy of Sciences, 266071, Qingdao, China aut Lu Douding The Second Institute of Oceanography, State Ocenic Administration, 310012, Hangzhou, China aut Xu Zhong Harbin Institute of Technology (Weihai), 264209, Weihai, China aut Yan Peishen Harbin Institute of Technology (Weihai), 264209, Weihai, China aut Acta Oceanologica Sinica The Chinese Society of Oceanography 32/2(2013-02-01), 66-75 0253-505X 32:2<66 2013 32 13131 https://doi.org/10.1007/s13131-013-0278-4 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s13131-013-0278-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Chen Guofu Harbin Institute of Technology (Weihai), 264209, Weihai, China aut NATIONALLICENCE 700 1- Wang Quanfu Harbin Institute of Technology (Weihai), 264209, Weihai, China aut NATIONALLICENCE 700 1- Zhang Chunyun Harbin Institute of Technology (Weihai), 264209, Weihai, China aut NATIONALLICENCE 700 1- Zhang Baoyu Institute of Oceanology, Chinese Academy of Sciences, 266071, Qingdao, China aut NATIONALLICENCE 700 1- Wang Guangce Institute of Oceanology, Chinese Academy of Sciences, 266071, Qingdao, China aut NATIONALLICENCE 700 1- Lu Douding The Second Institute of Oceanography, State Ocenic Administration, 310012, Hangzhou, China aut NATIONALLICENCE 700 1- Xu Zhong Harbin Institute of Technology (Weihai), 264209, Weihai, China aut NATIONALLICENCE 700 1- Yan Peishen Harbin Institute of Technology (Weihai), 264209, Weihai, China aut NATIONALLICENCE 773 0- Acta Oceanologica Sinica The Chinese Society of Oceanography 32/2(2013-02-01), 66-75 0253-505X 32:2<66 2013 32 13131 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer