Aim To study isolation and purification of Gonyautoxins from two differentstrains of Alexandrium minutum Halim. Methods Ethanol extracts of the two culture strains ofAlexandrium minutum Halim were fractionated by gel ...Aim To study isolation and purification of Gonyautoxins from two differentstrains of Alexandrium minutum Halim. Methods Ethanol extracts of the two culture strains ofAlexandrium minutum Halim were fractionated by gel filtration and ion exchange chromatography toobtain gonyautoxins. The toxin profiles were tested by HPLC method. Results Both strains containgonyautoxin-1, gonyautoxin-2, gonyautoxin-3 and gonyautoxin-4. The total gonyautoxin content ofAmtk4 strain is almost 10 times higher than that of Amtk2 strain. Conclusion Alexandrium minutumHalim Amtk4 strain is suitable for the preparation of Gonyautoxins.展开更多
Alexandrium minutum from the China Sea produces a range of toxins and causes damage to the local ecosystems and aquaculture.This is essential to understand environmental factors affecting potential distribution.Potent...Alexandrium minutum from the China Sea produces a range of toxins and causes damage to the local ecosystems and aquaculture.This is essential to understand environmental factors affecting potential distribution.Potential distributions of A.minutum in the China Sea were predicted based on maximum entropy modeling,and dominant environmental variables were studied through analyses of variable contributions and response curves.The results showed that highly suitable areas were mainly located in the southwest of the Yellow Sea,the Laizhou Bay,and north of Haizhou Bay.The coast of the South China Sea was predicted as a low-suitability area,and the coast of the East China Sea as an unsuitable area.Mean temperature of the coldest month(T_min)had the largest drop in permutation importance but a low percent contribution.The probability of presence of A.minutum increased with increasing concentration of nitrate(NO3−)and annual mean temperature(T_ann)over a wide range of them.The response curves decreased with increasing concentration of phosphate(PO43−)and ratio of NO_(3)^(−)to PO_(4)^(3−)(N_P_ratio)when PO_(4)^(3)−is above 0.049μmolL^(-1) and N_P_ratio above 4,indicating that low values of PO_(4)^(3−) concentration and N_P_ratio favour the occurrence of A.minutum.As a predictor,the variance of annual temperature(T_Var)had the highest percent contribution and gains.PO_(4)^(3−) was predicted to have much more information than the other variables,and exhibited the second largest drop in permutation importance and percent contribution.The T_Var and PO_(4)^(3−) are the most important dominant predictor variables.展开更多
The dinoflagellate Alexandrium minutum is often associated with harmful algal blooms (HABs). This species consists of many strains that differ in their ability to produce toxins but have similar morphology, making ide...The dinoflagellate Alexandrium minutum is often associated with harmful algal blooms (HABs). This species consists of many strains that differ in their ability to produce toxins but have similar morphology, making identification difficult. In this study, species-specific rRNA probes were designed for whole-cell fluorescence in situ hybridization (FISH) to distinguish A. minutum from two phylogenetic clades. We acquired the complete SSU to LSU rDNA sequences (GenBank accession numbers JF906989-JF906999) of 11 Alexandrium strains and used these to design rRNA targeted oligonucleotide probes. Three ribotype-specific probes, M-GC-1, M-PC-2, and M-PC-3, were designed. The former is specific for the GC clade ("Global clade") of A. minutum, the majority of which have been found non-toxic, and the latter two are specific for the PSP (paralytic shellfish poisoning)-producing PC clade ("Pacific clade"). The specificity of these three probes was confirmed by FISH. All cells in observed fields of view were fluorescently labeled when probes and target species were incubated under optimized FISH conditions. However, the accessibility of rRNA molecules in ribosomes varied among the probe binding positions. Thus, there was variation in the distribution of positive signals in labeled cells within nucleolus and cytosol (M-GC-1, M-PC-3), or just nucleolus (M-PC-2). Our results provide a methodological basis for studying the biogeography and population dynamics of A. minutum, and providing an early warning of toxic HABs.展开更多
A loop-mediated isothermal amplification （LAMP） assay was designed and evaluated for rapid de- tection of the toxic microalgae Alexandrium catenella and A. minutum, which can produce paralytic shellfish poisoning （...A loop-mediated isothermal amplification （LAMP） assay was designed and evaluated for rapid de- tection of the toxic microalgae Alexandrium catenella and A. minutum, which can produce paralytic shellfish poisoning （PSP）. Two sets of four specific primers targeting these two species were derived from the sequence of internal transcribed spacer （ITS） of ribosomal DNA. The method worked well in less than an hour under isothermal conditions of 65℃. LAMP specificity was validated in closely related algae as a comparison, suggesting the strict specificity of the LAMP primers. Two visual inspection approaches were feasible to interpret the positive or negative results. The detection lim- its of A. catenella and A. minutum samples using the LAMP assay were found to be 5.6 and 4.5 pg DNA, respectively. The sensitivity of this LAMP assay was 10 or 100-fold higher than Polymerase Chain Reaction （PCR） method in detecting the two microalgae. These characteristics of species specificity, sensitivity, and rapidity suggest that this method has the potentiality in the monitoring of red tide caused by A. catenella and A. minutum.展开更多
文摘Aim To study isolation and purification of Gonyautoxins from two differentstrains of Alexandrium minutum Halim. Methods Ethanol extracts of the two culture strains ofAlexandrium minutum Halim were fractionated by gel filtration and ion exchange chromatography toobtain gonyautoxins. The toxin profiles were tested by HPLC method. Results Both strains containgonyautoxin-1, gonyautoxin-2, gonyautoxin-3 and gonyautoxin-4. The total gonyautoxin content ofAmtk4 strain is almost 10 times higher than that of Amtk2 strain. Conclusion Alexandrium minutumHalim Amtk4 strain is suitable for the preparation of Gonyautoxins.
基金supported by the National Key Research and the Development Program of China(No.2019YFE 0124700)the China National Key Research and Development Program(No.2022YFC3106002)+1 种基金the National Natural Science Foundation of China(No.U1901215)the Startup Foundation for Introducing Talent of NUIST(No.2020r028).
文摘Alexandrium minutum from the China Sea produces a range of toxins and causes damage to the local ecosystems and aquaculture.This is essential to understand environmental factors affecting potential distribution.Potential distributions of A.minutum in the China Sea were predicted based on maximum entropy modeling,and dominant environmental variables were studied through analyses of variable contributions and response curves.The results showed that highly suitable areas were mainly located in the southwest of the Yellow Sea,the Laizhou Bay,and north of Haizhou Bay.The coast of the South China Sea was predicted as a low-suitability area,and the coast of the East China Sea as an unsuitable area.Mean temperature of the coldest month(T_min)had the largest drop in permutation importance but a low percent contribution.The probability of presence of A.minutum increased with increasing concentration of nitrate(NO3−)and annual mean temperature(T_ann)over a wide range of them.The response curves decreased with increasing concentration of phosphate(PO43−)and ratio of NO_(3)^(−)to PO_(4)^(3−)(N_P_ratio)when PO_(4)^(3)−is above 0.049μmolL^(-1) and N_P_ratio above 4,indicating that low values of PO_(4)^(3−) concentration and N_P_ratio favour the occurrence of A.minutum.As a predictor,the variance of annual temperature(T_Var)had the highest percent contribution and gains.PO_(4)^(3−) was predicted to have much more information than the other variables,and exhibited the second largest drop in permutation importance and percent contribution.The T_Var and PO_(4)^(3−) are the most important dominant predictor variables.
基金Supported by the National Natural Science Foundation of China(No.41006082)the Postdoctoral Science Foundation(No.20090461272)the National Natural Science Foundation of China for Creative Research Groups(No.40821004)
文摘The dinoflagellate Alexandrium minutum is often associated with harmful algal blooms (HABs). This species consists of many strains that differ in their ability to produce toxins but have similar morphology, making identification difficult. In this study, species-specific rRNA probes were designed for whole-cell fluorescence in situ hybridization (FISH) to distinguish A. minutum from two phylogenetic clades. We acquired the complete SSU to LSU rDNA sequences (GenBank accession numbers JF906989-JF906999) of 11 Alexandrium strains and used these to design rRNA targeted oligonucleotide probes. Three ribotype-specific probes, M-GC-1, M-PC-2, and M-PC-3, were designed. The former is specific for the GC clade ("Global clade") of A. minutum, the majority of which have been found non-toxic, and the latter two are specific for the PSP (paralytic shellfish poisoning)-producing PC clade ("Pacific clade"). The specificity of these three probes was confirmed by FISH. All cells in observed fields of view were fluorescently labeled when probes and target species were incubated under optimized FISH conditions. However, the accessibility of rRNA molecules in ribosomes varied among the probe binding positions. Thus, there was variation in the distribution of positive signals in labeled cells within nucleolus and cytosol (M-GC-1, M-PC-3), or just nucleolus (M-PC-2). Our results provide a methodological basis for studying the biogeography and population dynamics of A. minutum, and providing an early warning of toxic HABs.
基金The Science and Technology Commission of Shanghai Municipality under contract Nos 06235810108DZ1980802 and 10JC1418600a special research fund for the national non-profit institutes (East China Sea Fisheries Research Institute) under contract Nos 2007M22 and 2007Z01
文摘A loop-mediated isothermal amplification （LAMP） assay was designed and evaluated for rapid de- tection of the toxic microalgae Alexandrium catenella and A. minutum, which can produce paralytic shellfish poisoning （PSP）. Two sets of four specific primers targeting these two species were derived from the sequence of internal transcribed spacer （ITS） of ribosomal DNA. The method worked well in less than an hour under isothermal conditions of 65℃. LAMP specificity was validated in closely related algae as a comparison, suggesting the strict specificity of the LAMP primers. Two visual inspection approaches were feasible to interpret the positive or negative results. The detection lim- its of A. catenella and A. minutum samples using the LAMP assay were found to be 5.6 and 4.5 pg DNA, respectively. The sensitivity of this LAMP assay was 10 or 100-fold higher than Polymerase Chain Reaction （PCR） method in detecting the two microalgae. These characteristics of species specificity, sensitivity, and rapidity suggest that this method has the potentiality in the monitoring of red tide caused by A. catenella and A. minutum.
