Partial rDNA sequences of Prorocentrum minimum and Takayama pulchella were amplified, cloned and sequenced, and these sequence data were deposited in the GenBank. Eight oligonucleotide probes （DNA probes） were desig...Partial rDNA sequences of Prorocentrum minimum and Takayama pulchella were amplified, cloned and sequenced, and these sequence data were deposited in the GenBank. Eight oligonucleotide probes （DNA probes） were designed based on the sequence analysis. The probes were employed to detect and identify P. minimum and T. pulchella in unialgal and mixed algal samples with a fluorescence in situ hybridization method using flow cytometry. Epifluorescence micrographs showed that these specific probes labeled with fluorescein isothiocyanate entered the algal cells and bound to target sequences, and the fluorescence signal resulting from whole-cell hybridization varied from probe to probe. These DNA probes and the hybridization protocol we developed were specific and effective for P. minimum and T. pulchella, without any specific binding to other algal species. The hybridization efficiency of different probes specific to P. minimum was in the order： PM18S02 PM28S02 〉 PM28S01 〉PM18S01, and that of the probes specific to T. pulcheUa was TP18S02 TP28S01 〉 TP28S02 〉TP18S01. The different hybridization efficiency of the DNA probes could also be shown in the fluorescent signals between the labeled and unlabeled cells demonstrated using flow cytometry. The DNA probes PM18S02, PM28S02, TP18S02 and TP28S01, and the protocol, were also useful for the detection of Mgae in natural samples.展开更多
The naked dinoflagellate Takayama acrotrocha was identified as responsible for a bloom in Shenzhen Bay,Guangdong,China,in early spring 2021.The identification was confirmed by light,scanning,and transmission electron ...The naked dinoflagellate Takayama acrotrocha was identified as responsible for a bloom in Shenzhen Bay,Guangdong,China,in early spring 2021.The identification was confirmed by light,scanning,and transmission electron microscopy and molecular data based on the LSU ribosomal DNA(rDNA)and ITS rDNA sequences.This is the first record of T.acrotrocha bloom in the South China Sea.The sulcus of T.acrotrocha was wide and extended onto the epicone as a short intrusion in general,sometime the intrusion was not apparent and some were finger-like.The apical groove was deeply sigmoid.The nucleus was large,ovoid to cup-shaped and occupied most of the epicone.A large,rounded pyrenoid surrounded by a starch sheath was located at the left side to the centre of the hypocone.Under epifluorescence illumination,a row of large vesicular knobs was observed on the upper border of the cingulum.The intraspecific morphological variabilities in the clonal cultures of T.acrotrocha were investigated carefully.Cells that share the same diagnostic characters used for the description of Takayama xiamenensis such as the finger-like sulcus,a large nucleus located in the epicone and the similar pyrenoid type were observed.The LSU rDNA sequences of T.acrotrocha and T.xiamenensis only differed in 3 base pairs(bp)for a sequence length of 673 bp(with a similarity of 99.55%).For these reasons,we propose T.xiamenensis as a junior synonym of T.acrotrocha.展开更多
基金The Fujian Provincial Government of China under contract No 2005YZ1018 the Xiamen Municipal Government of China under contract No 3502Z20041059+4 种基金 the China Postdoctoral Science Foundation under contract No 20060400854the Open Fund of the State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences under contract No 2008FB005 the Specialized Research Fund for the Doctoral Program of Higher Education of China under contract 20070504076 the Open Fund of the Key Laboratory of Freshwater Fish Germplasm and Biotechnology of Ministry of Agriculture, Chinese Academy of Fishery Sciences under contract No LFB20070611the National Natural Science Foundation of China under contract No 40576055
文摘Partial rDNA sequences of Prorocentrum minimum and Takayama pulchella were amplified, cloned and sequenced, and these sequence data were deposited in the GenBank. Eight oligonucleotide probes （DNA probes） were designed based on the sequence analysis. The probes were employed to detect and identify P. minimum and T. pulchella in unialgal and mixed algal samples with a fluorescence in situ hybridization method using flow cytometry. Epifluorescence micrographs showed that these specific probes labeled with fluorescein isothiocyanate entered the algal cells and bound to target sequences, and the fluorescence signal resulting from whole-cell hybridization varied from probe to probe. These DNA probes and the hybridization protocol we developed were specific and effective for P. minimum and T. pulchella, without any specific binding to other algal species. The hybridization efficiency of different probes specific to P. minimum was in the order： PM18S02 PM28S02 〉 PM28S01 〉PM18S01, and that of the probes specific to T. pulcheUa was TP18S02 TP28S01 〉 TP28S02 〉TP18S01. The different hybridization efficiency of the DNA probes could also be shown in the fluorescent signals between the labeled and unlabeled cells demonstrated using flow cytometry. The DNA probes PM18S02, PM28S02, TP18S02 and TP28S01, and the protocol, were also useful for the detection of Mgae in natural samples.
基金Supported by the National Natural Science Foundation of China(Nos.41906112,42076144,42176201)the Science&Technology Basic Resources Investigation Program of China(No.2018FY100200)the Analysis Program on Key factors for Red Tide Forecast in Shenzhen。
文摘The naked dinoflagellate Takayama acrotrocha was identified as responsible for a bloom in Shenzhen Bay,Guangdong,China,in early spring 2021.The identification was confirmed by light,scanning,and transmission electron microscopy and molecular data based on the LSU ribosomal DNA(rDNA)and ITS rDNA sequences.This is the first record of T.acrotrocha bloom in the South China Sea.The sulcus of T.acrotrocha was wide and extended onto the epicone as a short intrusion in general,sometime the intrusion was not apparent and some were finger-like.The apical groove was deeply sigmoid.The nucleus was large,ovoid to cup-shaped and occupied most of the epicone.A large,rounded pyrenoid surrounded by a starch sheath was located at the left side to the centre of the hypocone.Under epifluorescence illumination,a row of large vesicular knobs was observed on the upper border of the cingulum.The intraspecific morphological variabilities in the clonal cultures of T.acrotrocha were investigated carefully.Cells that share the same diagnostic characters used for the description of Takayama xiamenensis such as the finger-like sulcus,a large nucleus located in the epicone and the similar pyrenoid type were observed.The LSU rDNA sequences of T.acrotrocha and T.xiamenensis only differed in 3 base pairs(bp)for a sequence length of 673 bp(with a similarity of 99.55%).For these reasons,we propose T.xiamenensis as a junior synonym of T.acrotrocha.
