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1.
Based on the sequence data of the nuclear ribosomal DNA internal transcribed spacer (ITS) 1, 5.8 S, and ITS 2, the molecular phylogeny was analyzed on Ulvaceae species collected from Qingdao coasts in summer of 2007, including 15 attached Ulva and Enteromorpha samples from 10 locations and 10 free-floating Enteromorpha samples from seven locations. The result supported the monophyly of all free-floating Enteromorpha samples, implying the unialgal composition of the free-floating Enteromorpha, and the attached Ulvaceae species from Qingdao coasts were grouped into other five clades, suggesting that they were not the biogeographic origin of the free-floating Enteromorpha in that season. Supported by NSFC (40506030); the Innovative Key Project of the Chinese Academy of Sciences (KZCX2-YW-209); Science & Technology Project of Qingdao City (06-2-2-12-JCH)  相似文献   

2.
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   

3.
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   

4.
An assessment with assistance of DNA barcoding was conducted on green macroalgae in coastal zone around Qingdao, China, during the period of April–December, 2011. Three markers were applied in molecular discrimination, including the plastid elongation factor tufA gene, the internal transcribed spacer (ITS) region of the ribosomal cistron and rubisco large subunit gene 3′ regions (rbcL-3P). DNA barcoding discriminated 8 species, excluding species of genus Cladophora and Bryopsis due to failures in amplification. We ascertained and corrected 4 species identified by morphological methods for effectively assisting the classification. The gene tufA presented more advantages as an appropriate DNA marker with the strongest amplification success rate and species discrimination power than the other two genes. The poorest sequencing success largely handicapped the application of ITS. Samples identified by tufA and rbcL as Ulva flexuosa were clustered into the clade of U. prolifera by ITS in the neighbor-joining tree. Confusion with discrimination of the complex of U. linza, U. procera and U. prolifera (as the LPP complex) still existed for the three DNA markers. Based on our results, rbcL is recommended as a preferred marker for assisting tufA to discriminate green macroalgae. In distinguishing green-tide-forming Ulva species, the free-floating sample collected from the green tide in 2011 was proved to be identical with U. prolifera in Yellow Sea for ITS and rbcL genes. This study presents a preliminary survey of green macroalgae distributed in the coastal area around Qingdao, and proves that DNA barcoding is a powerful tool for taxonomy of green macroalgae.  相似文献   

5.
Ulvacean green seaweeds are common worldwide; they formed massive green tides in the Yellow Sea in recent years, which caused marine ecological problems as well as a social issue. We investigated two major genera of the Ulvaceae, Ulva and Enteromorpha, and collected the plastid rbcL and nuclear ITS sequences of specimens of the genera in two sides of the Yellow Sea and analyzed them. Phylogenetic trees of rbcL data show the occurrence of five species of Enteromorpha (E. compressa, E. flexuosa, E. intestinalis, E. linza and E. prolifera) and three species of Ulva (U. pertusa, U. rigida and U. ohnoi). However, we found U. ohnoi, which is known as a subtropical to tropical species, at two sites on Jeju Island, Korea. Four ribotypes in partial sequences of 5.8S rDNA and ITS2 from E. compressa were also found. Ribotype network analysis revealed that the common ribotype, occurring in China, Korea and Europe, is connected with ribotypes from Europe and China/Japan. Although samples of the same species were collected from both sides of the Yellow Sea, intraspecific genetic polymorphism of each species was low among samples collected worldwide.  相似文献   

6.
Sargassum muticum is one of the most abundant and conspicuous native macroalgae species on the northern coasts of China. It often forms large-scale seaweed beds in subtidal zones. This investigation was designed to understand the intraspecific genetic relationships of this alga based on samples collected from four northern coastal sites of China, and to evaluate gamete release and growth capacity in laboratory conditions. The nuclear internal transcribed spacer 2 (ITS2) sequences of 16 samples from four locations were identical. Based on cox3 gene and partial rbcLS operon sequences, intraspecific genetic variability was detected with three and two ribotypes, respectively. Temperature, not irradiance, was shown to significantly affect gamete release and fertilization. Elevated temperature and irradiance enhanced the growth of germlings and vegetative branchlets. Maximum growth rate of germlings was detected at 18-24℃and an irradiance of 60-100 μmol photons/(m^2.s). Under ambient conditions (12-25℃ and 60-125 gmol photons/ (m2.s)), relative growth rate of young branchlets could reach 7.5%/d.  相似文献   

7.
The presence of Biecheleria cincta (=Woloszynskia cincta) in the Chinese coasts is reported for the first time. In scanning electron microscope, three to five series of vesicles and an elongated apical vesicle (EAV) were visible in the epicone, and both the hypocone and the cingulum had three series of vesicles each. Thin sections revealed that B. cincta possesses stalked pyrenoids and an unusual eyespot consisting of a stack of cisternae with brick-like materials (type E), thus supporting its transfer from Woloszynskia to Biecheleria. Spiny cysts formed spontaneously in culture, with an encystment rate of around 20%. Both large subunit ribosomal DNA (LSU rDNA) and internal transcribed spacer region (ITS) sequences in 12 strains from the Chinese coasts were determined. Phylogenetic analyses based on LSU rDNA and ITS sequences using Bayesian inference and maximum likelihood revealed two distinct ribotypes (referred to as ribotype A and B) in B. cincta. ITS region pairwise distances within B. cincta ranged from 0.024 to 0.072, suggesting the existence of a complex of cryptic species.  相似文献   

8.
Wang  Jianyan  Cen  Jingyi  Li  Si    Songhui  Moestrup  Øjvind  Chan  Kin-Ka  Jiang  Tao  Lei  Xiangdong 《中国海洋湖沼学报》2018,36(6):2202-2215
The dinoflagellate genus K arenia is known for recurrent harmful blooms worldwide. However, species diversity of the genus is generally overlooked owing to the difficulty of identifying small unarmored dinoflagellates. We have established four clonal cultures of K arenia longicanalis isolated from the type locality, Hong Kong harbor(strain HK01) and other three locations along the Chinese coasts(strains YB01, DT01, and NJ01). The morphology of the strain was studied by light and scanning electron microscopy(LM and SEM) and the pigment composition analyzed by high-performance liquid chromatography. We provide the first molecular data of K. longicanalis based on the large subunit(LSU) rRNA gene sequence and internal transcribed spacer(ITS). The four strains showed identical LSU rDNA sequences with a similarity of 99.4% to the holotype of K arenia umbella(strain KUTN05) from Australia. In the ITS phylogeny, the sequence of K. umbella branched between the Chinese strains of K. longicanalis. A careful comparison of the morphology of K. longicanalis and K. umbella reveals the similarity in the diagnostic characters. Diff erences may appear due to the sample treatment for SEM. We conclude that K. umbella is a junior synonym of K. longicanalis.  相似文献   

9.
Batrachospermum arcuatum Kylin is typically dioecious,although some monoecious specimens have been collected from four locations in North China.In this study,B.arcuatum populations,including monoecious and dioecious thalli,were collected from seven stream segments.The nuclear DNA internal transcribed spacer(ITS)region was sequenced for the seven populations,and a phylogenetic tree was constructed using Bayesian inference to assess intraspecifi c relationships.A haplotype network was also created.The ITS region in B.arcuatum from North China comprised 822–853 bp,with 10 haplotypes detected from the seven locations.The results of this study support the inclusion of monoecious individuals in the species B.arcuatum.  相似文献   

10.
This study is part of the endeavor to construct a comprehensive DNA barcoding database for common seaweeds in China. Identifications of red seaweeds, which have simple morphology and anatomy, are sometimes difficult solely depending on morphological characteristics. In recent years, DNA barcode technique has become a more and more effective tool to help solve some of the taxonomic difficulties. Some DNA markers such as COI (cytochrome oxidase subunitⅠ) are proposed as standardized DNA barcodes for all seaweed species. In this study, COI, UPA (universal plastid amplicon, domain V of 23S rRNA), and ITS (nuclear internal transcribed spacer) were employed to analyze common species of intertidal red seaweeds in Qingdao (119.3 -121 E, 35.35 -37.09 N). The applicability of using one or a few combined barcodes to identify red seaweed species was tested. The results indicated that COI is a sensitive marker at species level. However, not all the tested species gave PCR amplification products due to lack of the universal primers. The second barcode UPA had effective universal primers but needed to be tested for the effectiveness of resolving closely related species. More than one ITS sequence types were found in some species in this investigation, which might lead to confusion in further analysis. Therefore ITS sequence is not recommended as a universal barcode for seaweeds identification.  相似文献   

11.
The marine macroalgae Enteromorpha prolifera was one of the main algal genera that occurred in the widespread green tides in Qingdao, China, during the summers of 2007, 2008 and 2010. It is thus a plentiful source of biomass and could be used as a biofuel. In this study, the pyrolytic characteristics and kinetics of E. prolifera were investigated using thermogravimetric analysis (TGA) method. Cornstalk and sawdust were used as comparisons. Pyrolytic characteristics were studied using TG-DTG (thermogravimetry-derivative thermogravimetry) curves. Three stages in the pyrolytic process were determined: dehydration, dramatic weight loss and slow weight loss. E. prolifera was pyrolyzed at a lower initial temperature than the two terrestrial biomass forms. The apparent activation energy values for the three types of biomass were calculated and the mechanism functions were determined using 16 different mechanism functions, frequently used in thermal kinetics analysis. Activation energy values varied with mechanism function and the range of activation energy values for E. prolifera, cornstalk, and sawdust were 25-50 kJ/mol, 60-90 kJ/mol and 120-155 kJ/mol, respectively. This indicates that E. prolifera has low thermal stability for pyrolysis and good combustion characteristics.  相似文献   

12.
In 2008, a green tide broke out before the sailing competition of the 29th Olympic Games in Qingdao. The causative species was determined to be Enteromorpha prolifera (Ulva prolifera O. F. Müller), a familiar green macroalga along the coastline of China. Rapid accumulation of a large biomass of floating U. prolifera prompted research on different aspects of this species. In this study, we constructed a nonnormalized cDNA library from the thalli of U. prolifera and acquired 10 072 high-quality expressed sequence tags (ESTs). These ESTs were assembled into 3 519 nonredundant gene groups, including 1 446 clusters and 2 073 singletons. After annotation with the nr database, a large number of genes were found to be related with chloroplast and ribosomal protein, GO functional classification showed 1 418 ESTs participated in photosynthesis and 1 359 ESTs were responsible for the generation of precursor metabolites and energy. In addition, rather comprehensive carbon fixation pathways were found in U. prolifera using KEGG. Some stress-related and signal transduction-related genes were also found in this study. All the evidences displayed that U. prolifera had substance and energy foundation for the intense photosynthesis and the rapid proliferation. Phylogenetic analysis of cytochrome c oxidase subunit I revealed that this green-tide causative species is most closely affiliated to Pseudendoclonium akinetum (Ulvophyceae).  相似文献   

13.
Environmental DNA(eDNA)metabarcoding has emerged as a potentially powerful tool to monitor invasive fish species.As an alternative(or complementary)tool for biodiversity monitoring,e DNA metabarcoding had been used to detect species in aquariums,which represents an important transit avenue for introducing non-indigenous species with high population densities.In this study,eDNA metabarcoding as well as morphological characterization were used to reveal the diversity of non-indigenous species in a large aquarium at Qingdao Underwater World.Environmental DNA metabarcoding of 14 water samples at five locations from the Big Water Tank detected 24 non-indigenous species and four putative non-indigenous operational taxonomic units(OTUs).In contrast,only 20 non-indigenous species were observed by morphological characterization.Some species undetected by morphological characterization,such as Oreochromis niloticus(Linnaeus,1758),are highly adaptable to various environments and/or have invaded preferred regions where they threaten native aquatic species.eDNA metabarcoding also detected seven local fishes that were not identified by morphological characterization.However,analysis of OTU diversity among stations and sample replications revealed that eDNA varied within and/or between stations.Increasing sampling effort as well as negative controls are required to increase the detection rate of species and to eliminate false-positive OTUs.  相似文献   

14.
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   

15.
1 Introduction Dinoflagellate Alexandrium has received intensive at-tention in recent years, as it is a valuable bioactive com-pound resource and related to harmful algal bloom (HAB)which has negative impact on marine ecology systemaquiculture, human health and tourism (Hallegraeff andBolch, 1992; Smayda, 1997; Vale and Antonia de MSampayo, 2001). However, not all of the species in genusAlexandrium produce toxin (Wendy et al., 2001), so rapididentification and clarification of the phyloge…  相似文献   

16.
From 2007 to 2017, large-scale green tides occurred every year in the Yellow Sea of China, and Ulva prolifera was the main species leading to the green tide. In this study, we used the Polymerase chain reaction and 3' Rapid-amplification of c DNA ends technique to amplify the nr DNA-LSU and IGS sequences in U. prolifera and one species of Blidingia. These techniques showed 3259 bp of nrD NA-LSU and 3388 bp of IGS in U. prolifera and 3282 bp nr DNA-LSU and 3059 bp IGS in Blidingia sp. At the same time, tandem repeats, short dyads, palindromic and multiple simple repeat sequences in the IGS sequence were found by analyzing the structure of the IGS sequence in U. prolifera and Blidingia sp. G + C contents of the IGS sequence in U. prolifera and Blidingia sp. were 52.42% and 53.09%, respectively. We divided the U. prolifera into two types according to the morphological characteristics. Although the specimens of U. prolifera from the Qingdao coastal area, Jiangsu coastal area and the Yellow Sea have different morphologies, their ITS and IGS sequences are almost identical. Therefore, the main species of the green tides in the Qingdao coastal area, Jiangsu coastal area and the Yellow Sea are the same and have the same origin.  相似文献   

17.
A harmful algae bloom (HAB) is a dense aggregation of algae in a marine or aquatic environment that can result in significant environmental problems. To forecast the occurrence of HAB, development of a rapid and precise detection method is urgently required. In this study, two Skeletonema costatum-like diatoms (SK-1 and SK-2), were identified morphologically under a light microscope, and detected using fluorescent in situ hybridization (FISH). Strain SK-1 was isolated from a frequently HAB affected area of the East China Sea, and strain SK-2 from an aquatic farm in Qingdao, China. Fluorescent DNA probes were designed that were complementary to the ITS sequence (including 5.8S rDNA) of strain SK-1. After hybridization, strong green fluorescence was observed in cells of strain SK-1 under an epifluorescence microscope; however, no such fluorescence was observed with strain SK-2, which indicates that probes hybridized only the DNA of the target strain, SK-1, in species-specific manner, and that the two strains do not belong to a same species. This finding was confirmed by ITS sequence analysis. The FISH technique used in this study was sensitive, simple, and rapid, and is a promising tool for detecting target HAB species in natural environments.  相似文献   

18.
Peritrich ciliates are highly diverse and can be important bacterial grazers in aquatic ecosystems. Morphological identifi cations of peritrich species and assemblages in the environment are time-consuming and expertise-demanding. In this study, two peritrich-specifi c PCR primers were newly designed to amplify a fragment including the internal transcribed spacer(ITS) region of ribosomal rDNA from environmental samples. The primers showed high specifi city in silico, and in tests with peritrich isolates and environmental DNA. Application of these primers in clone library construction and sequencing yielded exclusively sequences of peritrichs for water and sediment samples. We also found the ITS1, ITS2, ITS, D1 region of 28 S rDNA, and ITS+D1 region co-varied with, and generally more variable than, the V9 region of 18 S rDNA in peritrichs. The newly designed specifi c primers thus provide additional tools to study the molecular diversity, community composition, and phylogeography of these ecologically important protists in dif ferent systems.  相似文献   

19.
Extremely large accumulation of green algae Enteromorpha prolifera floated along China’coastal region of the Yellow Sea ever since the summer of 2008.Amplified Fragment Length Polymorphism (AFLP) analysis was applied to assess the genetic diversity and relationships among E. prolifera samples collected from 9 affected areas of the Yellow Sea.Two hundred reproducible fragments were generated with 8 AFLP primer combinations,of which 194 (97%) were polymorphic. The average Nei’s genetic diversity, the coefficiency of genetic differentiation (Gst), and the average gene flow estimated from Gst in the 9 populations were 0.4018, 0.6404 and 0.2807 respectively. Cluster analysis based on the unweighed pair group method with arithmetic averages (UPGMA) showed that the genetic relationships within one population or among different populations were all related to their collecting locations and sampling time. Large genetic differentiation was detected among the populations.The E. prolifera originated from different areas and were undergoing a course of mixing.  相似文献   

20.
The rbcL gene of Ulva pertusa,Enteromorpha prolifera and Monostroma grevillei was amplified,sequenced and analyzed.By comparing the rbcL sequences with seven other Ulvales species retrieved from GenBank,the sequence divergences and the phyletic evolution were analyzed and the phylogenetic tree was constructed.From the phylogenetic tree,it can be found that U.pertusa,E.prolifera and U.californica group in one branch,while E.compressa,U.rigida and U.fenestrata cluster in another clade.Obviously,unlike the Enteomorpha species,the Ulva species do not gather in one branch.Therefore Ulva and Enteomorpha might be affiliates of one genus.E.compressa and E.intestinalis gathered together,which coincided with the morphological characters.However,the thallus of U.pertusa is thick and with many holes,which is different from E.prolifera in morphology.They cluster together in the phylogenetic tree with a genetic distance of 0.005.The results indicate that Ulva and Enteromorpha are not distinguished strictly.  相似文献   

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