为了对从猫粪便样品中分离的钩虫虫卵进行种类鉴定,本研究提取该虫卵基因组DNA,根据核糖体内转录间区1(ITS1)的保守序列设计钩虫属引物,通过PCR扩增、克隆、测序、相关软件分析,建立系统进化树,从分子水平对其进行种类鉴定。结果表明扩...为了对从猫粪便样品中分离的钩虫虫卵进行种类鉴定,本研究提取该虫卵基因组DNA,根据核糖体内转录间区1(ITS1)的保守序列设计钩虫属引物,通过PCR扩增、克隆、测序、相关软件分析,建立系统进化树,从分子水平对其进行种类鉴定。结果表明扩增出404 bp DNA片段,测序结果显示该片段包括309 bp ITS1和95 bp的5.8S rDNA,与NCBI中登录的序列进行比对以及采用Clustal X和MEGA5.1软件分析确定该钩虫为锡兰钩虫。这是我国首次建立钩虫的分子鉴定方法,也是40年来再次发现猫源锡兰钩虫,为锡兰钩虫分子生物学和分子流行病学研究奠定了基础。展开更多
Miamiensis avidus Thompson & Moewus,1964,is a cosmopolitan and well-known marine pathogenic ciliated protist.However,the taxonomy of this species up to now has remained controversial,especially with respect to the...Miamiensis avidus Thompson & Moewus,1964,is a cosmopolitan and well-known marine pathogenic ciliated protist.However,the taxonomy of this species up to now has remained controversial,especially with respect to the validity of the morphologically similar species,Philasterides dicentrarchi,which was considered as a junior synonym of M.avidus.In this study,a population of M.avidus was collected from the skin of pharaoh cuttlefish(Sepia pharaonis) cultured near the East China Sea,Ningbo,China and its morphology and phylogeny were investigated in detail based on living characters,infraciliature,small subunit(SSU) r DNA and ITS1-5.8 S-ITS2 region sequences.In addition,the morphometrics of a previously reported free-living population,collected from the Bohai Sea,were rechecked and analyzed.We compared the present two isolates with all historic populations of M.avidus and P.dicentrarchi,and found that their morphological characters were either highly similar or exactly identical,indicating that they are the same morphospecies.However,the phylogenetic analyses based on SSU r DNA or ITS1-5.8 S-ITS2 region sequences revealed that most M.avidus and P.dicentrarchi populations formed one clade,and the two isolates of M.avidus from Weifang and American Type Culture Collection clustered in another clade,which indicated that there might be cryptic species in Miamiensis avidus.展开更多
Species of the fungus Trichoderma, a genus of Hyphomycetes, are ubiquitous in the environment, but especially in soil. They have been used in a wide range of commercial applications including the production of hydrola...Species of the fungus Trichoderma, a genus of Hyphomycetes, are ubiquitous in the environment, but especially in soil. They have been used in a wide range of commercial applications including the production of hydrolases and in the biological control of plant diseases. A fundamental part of the Trichoderma antifungal system consists of a series of genes coding for a surprising variety of extracellular cell wall degrading enzymes (CWDE). Characterisation and identification of strains at the species level is the first step in utilizing the full potential of fungi in specific applications. One aim when isolating Trichoderma strains is to identify those which can be used in new agricultural and industrial applications. In the past it was not uncommon that biocontrol strains were defined as T. harzianum Rifai, due to the limited classification system of the genus Trichoderma. In recent years, several PCR-based molecular techniques have been used to detect and discriminate among microorganisms. Sequence analysis of the ITS regions of the ribosomal DNA and gene fragments as those corresponding to tef1 gene have been helpful in the neotypification, description and characterization of species in the genus Trichoderma. Another useful method for the identification of Trichoderma strains is the randomly amplified polymorphic DNA (RAPD) technique. Isozyme polymorphisms evaluation of five putative extracellular lytic enzymes loci (β-1,3-glucanase, β-1,6-glucanase, cellulase, chitinase and protease antivities) were carried out using representative strains of defined molecular groups. CWDE groupings obtained from biocontrol strains are discussed in relation to their phylogenetic location and antifungal activities. Compiling morphological, biochemical and sequence information data into a common database would provide a useful resource that could be used to accurately name new haplotypes identified in the future and correctly place them within the genus Trichoderma.展开更多
文摘为了对从猫粪便样品中分离的钩虫虫卵进行种类鉴定,本研究提取该虫卵基因组DNA,根据核糖体内转录间区1(ITS1)的保守序列设计钩虫属引物,通过PCR扩增、克隆、测序、相关软件分析,建立系统进化树,从分子水平对其进行种类鉴定。结果表明扩增出404 bp DNA片段,测序结果显示该片段包括309 bp ITS1和95 bp的5.8S rDNA,与NCBI中登录的序列进行比对以及采用Clustal X和MEGA5.1软件分析确定该钩虫为锡兰钩虫。这是我国首次建立钩虫的分子鉴定方法,也是40年来再次发现猫源锡兰钩虫,为锡兰钩虫分子生物学和分子流行病学研究奠定了基础。
基金supported by the National Natural Science Foundation of China(No.31572230)the Open Fund of Zhejiang Provincial Top Key Discipline of Aqua-culture in Ningbo University(No.xkzsc1417)+1 种基金the K.C.Wong Magna Fund in Ningbo Universitythe Scientific Research Foundation of Graduate School of Ningbo University
文摘Miamiensis avidus Thompson & Moewus,1964,is a cosmopolitan and well-known marine pathogenic ciliated protist.However,the taxonomy of this species up to now has remained controversial,especially with respect to the validity of the morphologically similar species,Philasterides dicentrarchi,which was considered as a junior synonym of M.avidus.In this study,a population of M.avidus was collected from the skin of pharaoh cuttlefish(Sepia pharaonis) cultured near the East China Sea,Ningbo,China and its morphology and phylogeny were investigated in detail based on living characters,infraciliature,small subunit(SSU) r DNA and ITS1-5.8 S-ITS2 region sequences.In addition,the morphometrics of a previously reported free-living population,collected from the Bohai Sea,were rechecked and analyzed.We compared the present two isolates with all historic populations of M.avidus and P.dicentrarchi,and found that their morphological characters were either highly similar or exactly identical,indicating that they are the same morphospecies.However,the phylogenetic analyses based on SSU r DNA or ITS1-5.8 S-ITS2 region sequences revealed that most M.avidus and P.dicentrarchi populations formed one clade,and the two isolates of M.avidus from Weifang and American Type Culture Collection clustered in another clade,which indicated that there might be cryptic species in Miamiensis avidus.
文摘Species of the fungus Trichoderma, a genus of Hyphomycetes, are ubiquitous in the environment, but especially in soil. They have been used in a wide range of commercial applications including the production of hydrolases and in the biological control of plant diseases. A fundamental part of the Trichoderma antifungal system consists of a series of genes coding for a surprising variety of extracellular cell wall degrading enzymes (CWDE). Characterisation and identification of strains at the species level is the first step in utilizing the full potential of fungi in specific applications. One aim when isolating Trichoderma strains is to identify those which can be used in new agricultural and industrial applications. In the past it was not uncommon that biocontrol strains were defined as T. harzianum Rifai, due to the limited classification system of the genus Trichoderma. In recent years, several PCR-based molecular techniques have been used to detect and discriminate among microorganisms. Sequence analysis of the ITS regions of the ribosomal DNA and gene fragments as those corresponding to tef1 gene have been helpful in the neotypification, description and characterization of species in the genus Trichoderma. Another useful method for the identification of Trichoderma strains is the randomly amplified polymorphic DNA (RAPD) technique. Isozyme polymorphisms evaluation of five putative extracellular lytic enzymes loci (β-1,3-glucanase, β-1,6-glucanase, cellulase, chitinase and protease antivities) were carried out using representative strains of defined molecular groups. CWDE groupings obtained from biocontrol strains are discussed in relation to their phylogenetic location and antifungal activities. Compiling morphological, biochemical and sequence information data into a common database would provide a useful resource that could be used to accurately name new haplotypes identified in the future and correctly place them within the genus Trichoderma.
