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Cobetia marina is a model proteobacteria in researches on marine biofouling. Its taxonomic nomenclature has been revised many times over the past few decades. ~To better understand the role of the surface-associated lifestyle of C. marina and the phylogeny of the family Halomonadaceae, we sequenced the entire genome of C. marina JCM 21022 ~T using single molecule real-time sequencing technology(SMR~T) and performed comparative genomics and phylogenomics analyses. ~The circular chromosome was 4 176 300 bp with an average GC content of 62.44% and contained 3 611 predicted coding sequences, 72 t RNA genes, and 21 r RNA genes. ~The C. marina JCM 21022 ~T genome contained a set of crucial genes involved in surface colonization processes. ~The comparative genome analysis indicated the significant diff erences between C. marina JCM 21022 ~T and Cobetia amphilecti KMM 296(formerly named C. marina KMM 296) resulted from sequence insertions or deletions and chromosomal recombination. Despite these diff erences, pan and core genome analysis showed similar gene functions between the two strains. ~The phylogenomic study of the family Halomonadaceae is reported here for the first time. We found that the relationships were well resolved among every genera tested, including Chromohalobacter, Halomonas, Cobetia, Kushneria, Zymobacter, and Halotalea.  相似文献   
2.
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.  相似文献   
3.
分析了几株自南海及东海分离的亚历山大藻的rDNA部分序列信息,其中包括核糖体大亚基(LSU)rDNA的5′端D1-D2区序列,以及5.8SrDNA和ITS区序列;同时也对实验室保种的部分来自其它国家和地区的亚历山大藻相关序列进行了测序和分析,并以此作为序列分析中的参考。采用ClustalX及MEGA2软件对所得到的序列信息进行了综合分析与对比。结果表明,分离自南海的塔玛亚历山大藻(Alexandriumtamarense)和分离自东海的链状亚历山大藻(A.catenella),即便是在ITS区和LSUrDNA等高变区,其序列信息也完全一致。与基因库中搜索到的其它亚历山大藻rDNA序列信息相比较,中国沿海的塔玛/链状亚历山大藻序列更接近于塔玛复合种的“亚洲温带”基因型。对于分离自南海的另外两株未定种的亚历山大藻,通过对比序列信息,发现它们与相关亚历山大藻(A.affine)非常接近。分离于我国台湾地区的微小亚历山大藻(A.minutum)在序列上与分离自新西兰的藻株相似,而与分离自欧洲的微小亚历山大藻藻株相差较大。中国沿海亚历山大藻rDNA序列信息的获得为针对有毒藻种设计特异性核酸探针,发展灵敏快速的生物检测技术奠定了基础。  相似文献   
4.
Previous studies found intraspecific diversity in Scrippsiella trochoidea A. R. Loeblich III, a widely distributed calcareous cyst-producing dinoflagellate. In this study, three strains (ST-1, ST-D6 and ST-K) of S. trochoidea isolated from the East Asian waters were studied, together with other geographical strains, to resolve their phylogenetic relationships. For the three East Asian isolates, two highly diverse regions of nuclear-encoded ribosomal DNA (rDNA), the 5.8S rDNA and its flanking internal transcribed spacers 1 and 2, and the 5' portion of the large-subunit rDNA (encompassing the "D1" and "D2" domains) were sequenced. Homologous sequences from other geographical isolates were selected from the GenBank database and the phylogenetic relationships were inferred from the molecular data of these strains. Strains of S. trochoidea were found to cluster into three major clades (STR1, STR2 and STR3), as reported in previous studies. Two of the three strains ST-1 and ST-K, were grouped in clade STR2, the other strain, ST-D6, belonged to clade STR3. The intraspecific diversity of S. trochoidea in East Asian waters makes it necessary to carry out phylogenetic investigations in combination with data of biogeography, population dynamics, and life cycle on the ecophysiology of a region.  相似文献   
5.
针对分离自南海的相关亚历山大藻,在进行核糖体大亚基DNA(LSU rDNA)全序列测序及比对的基础上,设计了针对其核糖体大亚基RNA(LSU rRNA)D10区的一个新的荧光素(FITC)标记寡核苷酸探针,并建立了相应的荧光原位杂交检测方法。在GenBank数据库中,BLAST检索的结果显示所设计的探针具有较高的特异性。杂交实验的结果也表明,这条探针对相关亚历山大藻的标记具有特异性,且标记效率可以达到100%,标记信号清晰明亮。本研究结果为相关亚历山大藻的监测及其种群动态研究提供了方法学依据,也为其它微藻特异性探针的设计提供了新的思路。  相似文献   
6.
用室内培养的塔玛亚历山大藻(Alexandrium tamarense ATHK藻株)作为实验对象,分别选择了5%福尔马林固定后常温保存、95%乙醇固定后常温保存、-20℃冷冻保存、鲁哥氏液(Lugols solution)固定后常温保存等方法,在5,15,30,60 d后对保存的样品进行单细胞PCR反应,并与新鲜样品进行比较。实验结果表明,用95%乙醇保存、-20℃冷冻保存、鲁哥氏液保存的样品在60 d后仍可扩增出目标条带,与新鲜样品没有差别,但是样品中的藻细胞形态有一些改变;而以5%福尔马林固定保存的样品只能在5 d后扩增出目标条带,但藻细胞形态比较完好。因此,对用于单细胞PCR实验的微藻样品,短期内(不多于5 d)可以采用福尔马林保存,而需要长期保存的样品,应当采用乙醇或鲁哥氏液固定,或者采用-20℃冷冻保存的方法。  相似文献   
7.
用室内培养的塔玛亚历山大藻(Alezandrium tamarense ATHK藻株)作为实验对象,分别选择了5%福尔马林固定后常温保存、95%乙醇固定后常温保存、-20℃冷冻保存、鲁哥氏液(Lugol’s solution)固定后常温保存等方法,在5,15,30,60d后对保存的样品进行单细胞PCR反应,并与新鲜样品进行比较。实验结果表明,用95%乙醇保存、-20℃冷冻保存、鲁哥氏液保存的样品在60d后仍可扩增出目标条带,与新鲜样品没有差别,但是样品中的藻细胞形态有一些改变;而以5%福尔马林固定保存的样品只能在5d后扩增出目标条带,但藻细胞形态比较完好。因此,对用于单细胞PCR实验的微藻样品,短期内(不多于5d)可以采用福尔马林保存,而需要长期保存的样品,应当采用乙醇或鲁哥氏液固定,或者采用-20℃冷冻保存的方法。  相似文献   
8.
Bangiales is the only order of the Bangiophyceae and has been suggested to be monophyletic. This order contains approximately 190 species and is distributed worldwide. Previous molecular studies have produced robust phylogenies among the red algae, but the divergence times, historical biogeography and evolutionary rates of Bangiales have rarely been studied. Phylogenetic relationships within the Bangiales were examined using the concatenated gene sets from all available organellar genomes. This analysis has revealed the topology((( Bangia, Porphyra) Pyropia) Wildemania). Molecular dating indicates that Bangiales diversifi ed approximately 246.40 million years ago(95% highest posterior density(HPD)= 194.78–318.24 Ma, posterior probability(PP)=0.99) in the Late Permian and Early Triassic, and that the ancestral species most likely originated from eastern Gondwanaland(currently New Zealand and Australia) and subsequently began to spread and evolve worldwide. Based on pairwise comparisons, we found a slower rate of nucleotide substitutions and lower rates of diversifi cation in Bangiales relative to Florideophyceae. Compared with Viridiplantae(green algae and land plants), the evolutionary rates of Bangiales and other Rhodophyte groups were found to be dramatically faster, by more than 3-fold for plastid genome(ptDNA) and 15-fold for mitochondrial genome(mtDNA). In addition, an average 2.5-fold lower dN/dS was found for the algae than for the land plants, which indicates purifying selection of the algae.  相似文献   
9.
Pyropia species grow in the intertidal zone and are cold-water adapted. To date, most of the information about the whole plastid and mitochondrial genomes(ptDNA and mtDNA) of this genus is limited to Northern Hemisphere species. Here, we report the sequencing of the ptDNA and mtDNA of the Antarctic red alga Pyropia endiviifolia using the Illumina platform. The plastid genome(195 784 bp, 33.28% GC content) contains 210 protein-coding genes, 37 tRNA genes and 6 rRNA genes. The mitochondrial genome(34 603 bp, 30.5% GC content) contains 26 protein-coding genes, 25 tRNA genes and 2 rRNA genes. Our results suggest that the organellar genomes of Py. endiviifolia have a compact organization. Although the collinearity of these genomes is conserved compared with other Pyropia species, the genome sizes show significant differences, mainly because of the different copy numbers of rDNA operons in the pt DNA and group II introns in the mtDNA. The other Pyropia species have 2–3 distinct intronic ORFs in their cox 1 genes, but Py. endiviifolia has no introns in its cox 1 gene. This has led to a smaller mtDNA than in other Pyropia species. The phylogenetic relationships within Pyropia were examined using concatenated gene sets from most of the available organellar genomes with both the maximum likelihood and Bayesian methods. The analysis revealed a sister taxa affiliation between the Antarctic species Py. endiviifolia and the North American species Py. kanakaensis.  相似文献   
10.
Previous studies found intraspecific diversity in Scrippsiella trochoidea A. R. Loeblich III, a widely distributed calcareous cyst-producing dinoflagellate. In this study, three strains (ST-1, ST-D6 and ST-K) of S. trochoidea isolated from the East Asian waters were studied, together with other geographical strains, to resolve their phylogenetic relationships. For the three East Asian isolates, two highly diverse regions of nuclear-encoded ribosomal DNA (rDNA), the 5.8S rDNA and its flanking internal transcribed spacers 1 and 2, and the 5′ portion of the large-subunit rDNA (encompassing the “D1“ and “D2” domains) were sequenced. Homologous sequences from other geographical isolates were selected from the GenBank database and the phylogenetic relationships were inferred from the molecular data of these strains. Strains of S. trochoidea were found to cluster into three major clades (STR1, STR2 and STR3), as reported in previous studies. Two of the three strains ST-1 and ST-K, were grouped in clade STR2, the other strain, ST-D6, belonged to clade STR3. The intraspecific diversity of S. trochoidea in East Asian waters makes it necessary to carry out phylogenetic investigations in combination with data of biogeography, population dynamics, and life cycle on the ecophysiology of a region.  相似文献   
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