development of a pna probe for fluorescence in situ hybridization detection of prorocentrum donghaiense发展机构的探针荧光原位杂交检测prorocentrum磷比.pdfVIP

Development of a PNA Probe for Fluorescence In Situ Hybridization Detection of Prorocentrum donghaiense 1,2,4 1 2 2,3 5 1 Guofu Chen , Chunyu Zhang *, Baoyu Zhang *, Guangce Wang *, Douding Lu , Zhong Xu , Peishen Yan1 1 State Key Laboratory of Urban Water Resource and Environment, Harbin Institute of Technology, Harbin, China, 2 Tianjin Key Laboratory of Marine Resources and Chemistry, Tianjin University of Science and Technology, Tianjin, China, 3 Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China, 4 The First Institute of Oceanography, SOA, Qingdao, China, 5 The Second Institute of Oceanography, SOA, Hangzhou, China Abstract Prorocentrum donghaiense is a common but dominant harmful algal bloom (HAB) species, which is widely distributed along the China Sea coast. Development of methods for rapid and precise identification and quantification is prerequisite for early-stage warning and monitoring of blooms due to P. donghaiense. In this study, sequences representing the partial large subunit rDNA (D1–D2), small subunit rDNA and internal transcribed spacer region (ITS-1, 5.8S rDNA and ITS-2) of P. donghaiense were firstly obtained, and then seven candidate DNA probes were designed for performing fluorescence in situ hybridization (FISH) tests on P. donghaiense. Based on the fluorescent intensity of P. donghaiense cells labeled by the DNA probes, the probe DP0443A displayed the best hybridization performance. Therefore, a PNA probe (PP0443A) analogous to DP0443A was used in the further study. The cells labeled with the PNA probe displayed more intensive green fluorescence than that labeled with its DNA analog. The PNA probe was used