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1.
Chlorophyta species are common in the southern and northern coastal areas of China. In recent years, frequent green tide incidents in Chinese coastal waters have raised concerns and attracted the attention of scientists. In this paper, we sequenced the 18S rDNA genes, the internal transcribed spacer (ITS) regions and the rbcL genes in seven organisms and obtained 536-566 bp long ITS sequences, 1 377-1 407 bp long rbcL sequences and 1 718-1 761 bp long partial 18S rDNA sequences. The GC base pair content was highest in the ITS regions and lowest in the rbcL genes. The sequencing results showed that the three Ulva prolifera (or U. pertusa) gene sequences from Qingdao and Nan’ao Island were identical. The ITS, 18S rDNA and rbcL genes in U. prolifera and U. pertusa from different sea areas in China were unchanged by geographic distance. U. flexuosa had the least evolutionary distance from U. californica in both the ITS regions (0.009) and the 18S rDNA (0.002). These data verified that Ulva and Enteromorpha are not separate genera.  相似文献   

2.
The wild Porphyra yezoensis collected from the Qingdao coast was used to prepare protoplasts by enzyme digestion. The pure line was constructed by cultivating the protoplasts. The 18S rDNA of the P. yezoensis pure line was cloned and sequenced. Sequence analysis was executed for this sequence and other 22 sequences retrieved from GenBank. A phylogenetic tree was constructed using the neighbor-joining method. The results revealed a high diversity of 18S rDNA sequences in genus Porphyra and the considerable variation of 18S rDNA sequences in different strains of the same species P. yezoensis and P. tenera. Significant difference of 18S rDNA sequence was observed between P. yezoensis from Qingdao, China, and the two strains of P. yezoensis from Japan, but the three strains of P. yezoensis formed a stable clade in the phylogenetic tree. These results indicate the possibility of interspecies and intraspecies discrimination of Porphyra using the 18S rDNA sequences.  相似文献   

3.
We collected nine Enteromorpha specimens from the coast of Yantai and evaluated their diversity based on analyses of their ITS (internal transcribed spacer) and 5S rDNA NTS (non-transcribed spacer) sequences. The ITS sequences showed slight nucleotide divergences between Enteromorpha linza and Enteromorpha prolifera. In contrast, multiple highly variable regions were found in the ITS region of Enteromorpha flexuosa. In general, there were more variable sites in the NTS region than in the ITS region in the three species. The variations in 5S rDNA NTS sequences indicated that the molecular diversity of Enteromorpha from the coast of Yantai is very high. However, a phylogenetic tree constructed using 5S rDNA NTS sequence data indicated that genetic differences were not directly related to geographical distribution.  相似文献   

4.
A newly identified bacterial disease of kelp(Saccharina japonica) gametophytes was found in clone cultures.It is characterized by swollen gametophyte cells in the early period of infection followed by filamentous fading.An alginolytic marine bacterium referred to as A-1 was isolated from the diseased gametophytes.On the basis of 16S rDNA sequencing and morphological,physiological and biochemical characteristics,the bacterium was identified as a strain of the genus Alteromonas.By testing Koch’s postulates,Alteromonas sp.A-1 was further confirmed as the pathogen.The infection process was also investigated using both scanning electron and light microscopy.  相似文献   

5.
The phylogenetic relationship among Chinese Parapenaeopsis species was studied by comparing 16S rDNA and COI gene sequence. The results showed that Parapenaeopsis hardwickii (Miers, 1878) and P. cultrirostris Alcock, 1906 identified from Chinese samples were the same species. Parapenaeopsis cornuta (Kishinouye, 1900), P. incisa Liu and Wang, 1987, and P. sinica Liu and Wang, 1987 were different species. However, their systematic relationship were closer than those of other species, and they formed an advanced branch in the phylogenetic trees. Additionally, the systematic position of P. hardwickii (Miers, 1878), P. hungerfordi Alcock, 1905, and P. tenella (Bate, 1888) were not stable in the phylogenetic trees. More genetic markers such as 18S rDNA and 28S rDNA were supposed to be used to confirm the phylogenic relationship of Parapenaeopsis species.  相似文献   

6.
The systematic classification of the Eucheumatoideae is difficult because of their variable morphology and interpretation of reproductive structures. Kappaphycus and Eucheuma specimens cultivated on the Hainan and Fujian coast of China were introduced from Vietnam, the Philippines and Indonesia. Combined with morphological characteristics, all Kappaphycus and Eucheuma cultivated strains were identified by internal transcribed spacer (ITS) sequences. The phylogenetic tree was constructed using neighbor-joining and maximum likelihood methods. The results indicate that different ITS sequence lengths occurred in the different genera and species. An obvious difference in morphology could be found in the protuberance shape between Kappaphycus and Eucheuma. The protuberance in Eucheuma was thorn-like and in Kappaphycus was wartlike or papillate. Their ITS sequence lengths differed significantly in nucleotide variation rates up to 58.55%-63.90%. All nucleotide variations occurred in the ITS1 and ITS2 regions except for five nucleotide transversions in the 5.8S rDNA region. In addition, the difference was at the branches among congeneric species. Kappaphycus sp. had branches with small buds, while K. alvarezii did not have such a feature. The nucleotide variation rates varied from 7.02% to 7.48% among species; within the same species of the clades it was <1.20%. Eucheumatoideae algae cultivated in China consisted of three clades, K. alvarezii, Kappaphycus sp., and E. denticulatum. The results indicate that ITS sequence analysis was an effective way for identification of interspecies and intraspecies phylogenetic relationships and might provide a clue for molecular identification of algal Eucheumatoideae.  相似文献   

7.
Green tides are caused by the proliferation of chlorophytes under suitable hydrographic conditions.These blooms lead to environmental degradation and negatively impact the waters and seagrass beds,as well as fishing and other recreational activities in the bay.A comprehensive ecological understanding of the bloom dynamics,including the origin and persistence,is needed to foster management decisions.The algae in the great majority of green tide blooms usually belong to two genera of Ulvophyceae,Ulva and Enteromorpha.Ulva has been observed more often in recent years.In China,green tides occurred for the first time in the middle area of the Yellow Sea in 2007,and a large-scale algae blooming broke out in the middle and southern areas of the Yellow Sea in late May 2008.We identified them as Ulva prolifera by comparative analysis of the rDNA internal transcribed spacer 1 (ITS1),5.8S and ITS2 sequences in combination with microscopic observation.Morphological differences were found between the free-floating algae and the attached thalli.Various reproduction patterns of the free-floating algae include sexual,asexual and vegetative propagations,which played important roles in the long-term green tide persistence in China.The ITS sequences of the blooming algae were identical to those of the samples from the Lianyungang sea area but were different from the attached samples from the Qingdao sea area.The results infer that the blooms are originated from other sea areas rather than from the local attached populations.  相似文献   

8.
During a survey for marine microalgal resources, we isolated a rare marine euglenoid from the coastal waters of Qingdao, China in 2009, and established a pure culture. Electron microscopic and molecular phylogenetic (18S rDNA and 16S rDNA sequences) analyses revealed a close affinity with Eutreptiella gymnastica, a bloom-forming species. Different culture conditions were monitored to understand optimal E. gymnastica growth characteristics. The optimal growth conditions in a batch culture of this isolate were 20°C, 160 μmol photons/(m2×s) of white light, and a salinity of 10-31. Nutrient experiments demonstrated that growth increased dramatically with a phosphorus concentration greater than 72 μmol/L. Understanding the effect of culture conditions on E. gymnastica may help understanding the blooming mechanism of this alga in its natural environment.  相似文献   

9.
The decomposition of a large amount of unexploited macroalgal resource along the coast of China often results in heavy environmental pollution. In order to pave a way of using macroalgae as the dietary ingredient of rabbitfish Siganus canaliculatus, one of a few farmed herbivorous marine teleosts in China, its preference (feeding selectivity) for different macroalgae was determined in this study. Seven seaweed species abundantly inhabiting the coast of east Guangdong Province were exposed simultaneously to rab-bitfish juveniles in laboratory (multiple-choice feeding) with their content and absolute intake assayed. It was found that the most preferred algae were Ulva prolifera, Gracilaria lemaneiformis and Chaetomorpha linum, less preferred algae were U. pertusa and Porphyra haitanensis, and least preferred ones were Sargassum fusiforme and Corallina sessilis. Such an order did not change when one to four relatively preferred seaweeds were removed. The preferred seaweeds were richer in protein and soluble sugar thus higher in energy than the least preferred. In addition, this fish was found to favor filamentous and flat algae rather than calcified ones. Ac-cordingly, the richness of nutrients and morphological characteristics determined the preference of S. canaliculatus for tested macro-algae.  相似文献   

10.
Seventy-eight marine fungal strains were isolated from sediment samples collected off the coast of Nanji Island,Wenzhou,Zhejiang Province,China.Antibacterial screening using the agar disc method showed that 19 of the isolated strains could inhibit at least one pathogenic Vibrio from Pseudosciaena crocea.Subsequent screening confirmed that nine strains produced antibacterial metabolites that had activity against one or several types of pathogenic Vibrio.Strain NJ0104 had the widest antimicrobial spectrum and strong activity,particularly against Vibrio parahaemolyticus-MM0810072.A preliminary study of NJ0104 antibacterial metabolites demonstrated that they had thermal stability up to 80°C,ultraviolet stability up to 40 min and pH stability between 4.0-7.0.In addition,the antibacterial metabolites were readily soluble in butanol.To identify the specific strain,the ITS-5.8S rDNA regions of NJ0104 were PCR amplified and sequenced.Based on the combination of phenotypic and genotypic data,the strain was identified as Arthrinium sp.  相似文献   

11.
Pelagic copepods play an important role in the marine food web. However, a full understanding of the ecological status of this zooplankton group depends on the careful study of their natural diets. In previous PCR-based copepod diet studies, we found many apostome ciliates that live symbiotically under the exoskeleton of the copepods, and their sequences were often over-represented in the 18S rRNA gene (18S rDNA) libraries. As a first step to address this issue, we designed three apostome ciliate 18S rDNA blocking primers, and tested their blocking efficiency against apostome ciliate 18S rDNA under various PCR conditions. Using a semi-quantitative PCR method, we optimized the conditions to efficiently amplify the 18S rDNA of the prey while simultaneously excluding the symbiotic apostome ciliates. This technique will facilitate PCR-based diet studies of copepods and other zooplankton in their natural environments.  相似文献   

12.
Colony morphology is important for Microcystis to sustain a competitive advantage in eutrophic lakes. The mechanism of colony formation in Microcystis is currently unclear. Extracellular polysaccharide (EPS) has been reported to play an important role in cell aggregate formation of some phytoplankton. Microcystis aeruginosa was cultivated under varied abiotic conditions, including different nutrient, light, and temperature conditions, to investigate their effects on EPS production and morphological change. The results show that nutrient concentration and light intensity have great effects on EPS production in M. aeruginosa. There was a considerable increase in EPS production after M. aeruginosa was cultivated in adjusted culture conditions similar to those present in the field (28.9mg C/L, 1.98mg N/L, 0.65mg P/L, light intensity:100 μmol/(m2s)). These results indicate that abiotic factors might be one of the triggers for colony formation in Microcystis.  相似文献   

13.
A molecular phylogenetic analysis of the genus Scomber was conducted based on mitochondrial (COI, Cyt b and control region) and nuclear (5S rDNA) DNA sequence data in multigene perspective. A variety of phylogenetic analytic methods were used to clarify the current taxonomic Classification and to assess phylogenetic relationships and the evolutionary history of this genus. The present study produced a well-resolved phylogeny that strongly supported the monophyly of Scomber. We confirmed that S. japonicus and S. colias were genetically distinct. Although morphologically and ecologically similar to S. colias, the molecular data showed that S. japonicus has a greater molecular affinity with S. australasicus, which conflicts with the traditional taxonomy. This phylogenetic pattern was corroborated by the mtDNA data, but incompletely by the nuclear DNA data. Phylogenetic concordance between the mitochondrial and nuclear DNA regions for the basal nodes Supports an Atlantic origin for Scomber. The present-day geographic ranges of the species were compared with the resultant molecular phylogeny derived from partition Bayesian analyses of the combined data sets to evaluate possible dispersal routes of the genus. The present-day geographic distribution of Scomber species might be best ascribed to multiple dispersal events. In addition, our results suggest that phylogenies derived from multiple genes and long sequences exhibited improved phylogenetic resolution, from which we conclude that the phylogenetic reconstruction is a reliable representation of the evolutionary history of Scomber.  相似文献   

14.
Acrochaete leptochaete (Huber) Nielsen (Chaetophoraceae, Chlorophyta) was isolated from the macroalgae Chaetomorpha collected from intertidal pools in Rongcheng, Shandong, China. 18S rDNA combined with ITS regions were used to ascertain the morphological identification of the isolated material. Based on the unialgal culture, asexual reproduction and growth characteristics of A. leptochaete were investigated over wide ranges of temperature and irradiance. Results revealed that asexual reproduction of A. leptochaete could be realized by biflagellate zoospores. The zoospores germinated directly to give self- replicating generations. Zoospore germination was bipolar. A temperature range from 13-21°C and a lower irradiance of 36 μmol/(m2·s) were most favorable for the growth of A. leptochaete. Thallus organization, an important taxonomic criterion for the genus Acrochaete, was affected markedly by temperature and irradiance. Our results extend the knowledge about the species’ general biology and its morphological plasticity. For classification and identification of a simple microphytic algae like A. leptochaete, which are traditionally placed in the class Chaetophoraceae, we propose that molecular tools associated with culture observations are applied.  相似文献   

15.
The morphological characteristics and the cuttlebone formation of Sepia esculenta exposed to different water temperature fluctuations were investigated under laboratory conditions. Temperature fluctuation cycles (15 cycles, 60 d in total) consisted of the following three regimes of 4 d duration: keeping water temperature in 26°C for 3 d (Group A), 2 d (Group B), 0 d (Group C, control); then keeping water temperature in 16°C for the next 1, 2, 4 d. No significant difference in the survival rate was observed between the control and temperature fluctuation groups (P>0.05). Lamellar depositions in a temperature fluctuation cycle were 2.45±0.02 for Group A, 2.00±0.02 for Group B, and 1.78±0.02 for Group C (P<0.05). The relationship between age and number of lamellas in the cuttlebone of S. esculenta under each water temperature fluctuation could be described as the linear model and the number of lamellas in the cuttlebone did not correspond to actual age. Group A had the highest cuttlebone growth index (CGI), the lowest locular index (LI), and inter-streak distances comparing with those of control group. However, the number of lamellas and LI or CGI showed a quadratic relationship for each temperature fluctuation group. In addition, temperature fluctuations caused the breakage of cuttlebone dark rings, which was considered a thermal mark. The position of the breakage in the dark rings was random. This thermal mark can be used as supplementary information for marking and releasing techniques.  相似文献   

16.
In this study, marine microplankton were identified by combining standard light microscopy with Sanger 18S rRNA gene sequencing. The image-matching individual PCR technique was applied to identify the image collectable unicellular microplankton to genera. Instead of pure strain culture and morphological identification, microplankton individual cells were isolated and fixed with glutaraldehyde, frozen and stored for months. Finally, they were imaged under a microscope and molecularly identified via phylogenetic analysis of their 18S ribosomal RNA gene(18S rDNA). Microplankton cells were collected at 30 locations in South China Sea, and were assigned to 21 known and 4 unidentified genera(2 uncultured fungi and 2 uncultured stramenopiles) with phylogenetic analysis in parallel to the morphological identification.  相似文献   

17.
Chinese shrimp (Fenneropenaeus chinensis) is an economically important aquaculture species in China. However, cytogenetic and genomic data is limited in the organism partly because the chromosomes are difficult to isolate and analyze. In this study, fluorescence in-situ hybridization (FISH) was used to identify the chromosomes of F. chinensis. The 5S ribosomal RNA gene (rDNA) of F. chinensis was isolated, cloned and then used as a hybridization probe. The results show that the 5S rDNA was located on one pair of homologous chromosomes in F. chinensis. In addition, triploid shrimp were used to evaluate the feasibility of chromosome identification using FISH and to validate the method. It was confirmed that 5S rDNA can be used as a chromosome-specific probe for chromosome identification in F. chinensis. The successful application of FISH in F. chinensis shows that chromosome-specific probes can be developed and this finding will facilitate further research on the chromosomes of penaeid shrimps.  相似文献   

18.
The use of a microwave-assisted extraction(MAE) method for the extraction of phlorotannins from Saccharina japonica Aresch(S.japonica) has been evaluated with particular emphasis on the influential parameters,including the ethanol concentration,solid/liquid ratio,extraction time,extraction temperature,and microwave power.The MAE procedure was optimized using single-factor design and orthogonal array design(OAD).The content of total phlorotannins in S.japonica was determined using a Folin-Ciocalteu(FC) assay.A maximum total phlorotannin content of 0.644 mg of phloroglucinol equivalent per gram of dry weight plant(mg PGE/g DW) was obtained using the optimized model,which included an ethanol concentration of 55%,solid/liquid ratio of 1:8,extraction time of 25 min,irradiation power of 400 W,and temperature of 60°C.Under similar conditions,the application of a conventional extraction method led to a lower phlorotannin yield of 0.585 mg PGE/g WD.These results demonstrated that the MAE approach provided better results for the extraction of phlorotannins from S.japonica and was a promising technique for the extraction of phenolic compounds from S.japonica and other materials.In addition,screening tests for the inhibitory activity showed that the phlorotannin-containing extracts significantly inhibited the growth of human hepatocellular carcinoma cells(HepG2) by inducing their apoptosis.The morphological changes that occurred during cell apoptosis were characterized using Hoechst33258 staining.  相似文献   

19.
We cultured different-sized fractions of dominant phytoplankton species, Skeletonema costatum, Chaetoceros curvisetus, and Thalassiosira nordenskiöldii, collected in different sea areas in various seasons, and measured and compared their C, N, P, Si contents. The N content of these species is similar, while the C, P, and Si contents of S. costatum from eutrophic Changjiang (Yangtze River) estuary are higher than those from Jiaozhou Bay (JZB), particularly the content of Si. The C, N, P, and Si contents of cultured phytoplankton in JZB increase with size fraction augmentation, and the percentages of C, N, and P follow the same trend, while the percentage of Si remain constant. Moreover, S. costatum from small-sized fraction assimilated Si more easily than C. curvisetus and T. nordenskiöldii, which is explained by the dominance of S. costatum under the conditions of low SiO3-Si concentration in JZB. The C, N, P, and Si contents of cultured S. costatum collected during summer and winter are higher, which is consistent with the phytoplankton blooming seasons in JZB. The SiO3-Si concentration of seawater during spring restrain the growth of phytoplankton, supported by the fact that the N, P, and Si contents and their ratios in cells of cultured S. costatum are low in spring season.  相似文献   

20.
Skeletonema tropicum is regarded as a species with an affinity to warm waters and it has never been reported in seas where temperatures drop below 11°C in winter. Previous studies in China reported that S. tropicum was restricted to subtropical and warm temperate seas (East and South China Seas), but the species was recently found during August cruises of 2009 and 2010 in Jiaozhou Bay, Yellow Sea, located several hundred kilometers to the north. Here, winter water temperatures often drop below 5°C. Identification of S. tropicum was confirmed under light and scanning electronic microscopes and maximum cell abundance in Jiaozhou Bay was estimated as 1.73×10 4 cell/L. This record of S. tropicum in Jiaozhou Bay represents a significant northward expansion in the geographic range of the species. Ship ballast water was identified as a possible carrier of S. tropicum from southern places along Chinese coastline, and in addition, thermal pollution from local power stations and seawater desalination plants may provide suitable conditions for species over-wintering.  相似文献   

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