Gel microbead cultivation with a subenrichment procedure can yield better bacterial cultivability from a seawater sample than standard plating method

A gel microbead (GMD) cultivation method was employed to cultivate microorganisms from an amphioxus breeding zone in Qingdao, P. R. China. The culture results were compared with those by standard plating method. In the GMD-based method, the microcolony-forming GMDs were sorted by fluorescence-activated cell sorting (FACS). To further get pure cultures, a subsequent enrichment culture and a streaking purification procedure were conducted on marine R2A medium. Eighty bacterial strains isolated by the GMD-based method were randomly selected for sequencing. These isolates belonged to Alphaproteobacteria (33%), Gammaproteobacteria (44%), Bacteroidetes (11%), Actinobacteria (5%), Firmicutes (5%), Epsilonproteobacteria (1%), and Verrucomicrobia (1%), the last two groups being usually difficult to culture. The 16S rRNA gene sequences revealed a diverse community with 91.1%-100% of the bacterial rRNAs similarities. Thirteen strains were sharing 16S rRNA gene sequence which was less than 97% similar to any other rRNA genes currently deposited in TYP16S database. Seventy isolates derived from the standard plating method fell into 4 different taxonomic groups: Alphaproteobacteria (9%), Gammaproteobacteria (81%), Bacteroidetes (7%) and Firmicutes (3%) with a 16S rRNA gene sequence similarities between 95.8%-100%, in which only 3 strains were sharing 16S rRNA gene sequence of less than 97%. The results indicated that the GMD-based method with subenrichment culture yielded more taxonomic groups and more novel microbial strains, including members of previously rarely cultured groups, when compared with the standard plating method, and that this method markedly improved the bacterial cultivability.     

Microbial diversity in the sediments of the southern Mariana Trench

Microbial diversity in the abyssal sediments beneath the seafloor of 30, 94, and 151 cm near the southern end of the Mariana Trench was analyzed in the Illumina HiSeq 2500 platform. Results show that the microbial populations were dominated by bacteria but merely no archaea were identified at the three depths. In the bacterial community,Proteobacteria and Firmicutes dominated the total taxon tags, followed by Bacteroidetes, Actinobacteria, Planctomycetes, Cyanobacteria, and Chloroflexi, which together account for over 99% of the total population. Similar to that in the seawater in the trench, the operational taxonomic units(OTUs) belonging to Gammaproteobacteria from the sediment samples showed high abundance.However, common bacterial OTUs in the water of the trench including Nitrospirae and Marinimicrobia were hardly found in the sediments from the southern Mariana Trench or the hadal region. Therefore, this study documented for the first time the compositions of microbial diversity in the trench sediments, revealed the difference in microbial diversity in water and sediment of the trench and will enrich the knowledge on the microbial diversity in the abyssal areas.     

Analysis of expressed sequence tags from the Ulva prolifera (Chlorophyta)

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).     

Antioxidant system responses in two co-occurring green-tide algae under stress conditions

Green tides have occurred every year from 2007 to 2014 in the Yellow Sea. Ulva prolifera(Müller) J. Agardh has been identified as the bloom-forming alga,co-occurring with U. intestinalis. We observed distinct strategies for both algal species during green tides. U. prolifera exhibited a high abundance initially and then decreased dramatically,while U. intestinalis persisted throughout. The antioxidant system responses of these two macroalgae were compared in the late phase of a green tide(in-situ) and after laboratory acclimation. Lipid peroxidation and antioxidant system responses differed significantly between the two. Malondialdehyde and hydrogen peroxide contents increased significantly in-situ in U. prolifera,but not in U. intestinalis. In U. prolifera,we observed a significant decrease in total antioxidant ability(T-AOC),antioxidant enzymes(SOD and Apx),and non-enzyme antioxidants(GSH and As A) in-situ. U. intestinalis showed the same pattern of T-AOC and SOD,but its Gpx,Apx,and GSH responses did not differ significantly. The results suggest that U. prolifera was more susceptible than U. intestinalis to the harsh environmental changes during the late phase of a Yellow Sea green tide. The boom and bust strategy exhibited by U. prolifera and the persistence of U. intestinalis can be explained by differences in enzyme activity and antioxidant systems.     

Bacterial community succession in response to dissolved organic matter released from live jellyfish

Jellyfish blooms have increased worldwide,and the outbreaks of jellyfish population not only affect the food web structures via voracious predation but also play an important role in the dynamics of nutrients and oxygen in planktonic food webs.However,it remains unclear whether specific carbon compounds released through j ellyfish metabolic processes have the potential to shape bacterial community composition.Therefore,in this study,we aimed to investigate the compositional succession of the bacterioplankton community in response to the dissolved organic matter(DOM)released by the live Scyphomedusae Cyanea lamarcki'i and Chrysaora hysoscella collected from Helgoland Roads of the North Sea.The bacterial community was significantly stimulated by the DOM released form live jellyfish and different dominant phylotypes were observed for these two Scyphomedusae species.Furthermore,the bacterial community structures in the different DOM sources,jellyfish-incubated media,Kabeltonne seawater,and artificial seawater(DOM-free)were significantly different,as revealed by automated ribosomal intergenic spacer analy sis fingerprints.Catalyzed reporter depo sition fluorescence in situ hybridization(CARD-FISH)revealed a rapid species-specific shift in bacterial community composition.Gammaproteobacteria dominated the community instead of the Bacteroidetes community for C.lamarckii,whereas Gammaproteobacteria and Bacteroidetes dominated the community for C.hysoscella.The significant differences in the bacterial community composition and succession indicate that the components of the DOM released by jellyfish might differ with jellyfish species.     

Impact of redox-stratification on the diversity and distribution of bacterial communities in sandy reef sediments in a microcosm

Relationships between microbial communities and geochemical environments are important in marine microbial ecology and biogeochemistry. Although biogeochemical redox stratification has been well documented in marine sediments, its impact on microbial communities remains largely unknown. In this study, we applied denaturing gradient gel electrophoresis (DGGE) and clone library construction to investigate the diversity and stratification of bacterial communities in redox-stratified sandy reef sediments in a microcosm. A total of 88 Operational Taxonomic Units (OTU) were identified from 16S rRNA clone libraries constructed from sandy reef sediments in a laboratory microcosm. They were members of nine phyla and three candidate divisions, including Proteobacteria (Alpha-, Beta-, Gamma-, Delta-, and Epsilonproteobacteria), Actinobacteria, Acidobacteria, Bacteroidetes, Chloroflexi, Cyanobacteria, Firmicutes, Verrucomicrobia, Spirochaetes, and the candidate divisions WS3, SO31 and AO19. The vast majority of these phylotypes are related to clone sequences from other marine sediments, but OTUs of Epsilonproteobacteria and WS3 are reported for the first time from permeable marine sediments. Several other OTUs are potential new bacterial phylotypes because of their low similarity with reference sequences. Results from the 16S rRNA, gene clone sequence analyses suggested that bacterial communities exhibit clear stratification across large redox gradients in these sediments, with the highest diversity found in the anoxic layer (15–25 mm) and the least diversity in the suboxic layer (3–5 mm). Analysis of the nosZ, and amoA gene libraries also indicated the stratification of denitrifiers and nitrifiers, with their highest diversity being in the anoxic and oxic sediment layers, respectively. These results indicated that redox-stratification can affect the distribution of bacterial communities in sandy reef sediments.     

Shifts of sediment bacterial community and respiration along a successional gradient in a typical karst plateau lake wetland(China)

Bacteria are important regulators of carbon cycling in lakes and are central to sediment ecosystem processes. However, the sediment microbial communities and their respiratory responses to the lake wetland succession are poorly understood. In this study, we collected sediment samples from four different succession points(the Potamogeton lucens zone, the Scirpus tabernaemontani zone, the Scirpus triqueter zone, and the Juncus effusus zone) in the Caohai Wetland of the Guizhou Plateau(China). The bacterial communities at these succession points were studied using a high-throughput sequencing approach. The sediment microbial respiration(SR) was measured using static chambers in the field and basal respiration(BR) was determined in the laboratory. The results show that the dominant bacterial taxa in the sediment was Proteobacteria(34.7%), Chloroflexi(17.8%), Bacteroidetes(7.3%), Acidobacteria(6.6%), and Cyanobacteria(6.1%). Principal coordinate analysis showed that the microbial community structure differs significantly at different sampling points along the successional gradient, indicating that the bacterial community structure is sensitive to the lake wetland succession. Different hydrological regimes and soil characteristics such as NH_4~+-N, Fe~(2+), Mn~(2+), and sediment organic carbon(SOC) content may be important factors responsible for the differences in the sediment microbial characteristics of the different successional stages in the Caohai wetland. Additionally, it was found that the SR increased significantly from the P. lucens zone to the J. effusus zone, but BR had the opposite response. The shifts in the bacterial community structure along the successional gradient may be the main reason for the observed differences in sediment respiration.     

The bacterial diversity and community composition altered in the oxygen minimum zone of the Tropical Western Pacific Ocean

The oxygen minimum zones(OMZs) are globally expanding,yet the variation pattern of microbial communities related to dissolved oxygen levels remain unclear.Spatial variability of bacterial diversity and community composition(repre sented by 16 S rRNA) of six stations was investigated within the water column in the seamount area of Tropical Western Pacific Ocean(TWPO) in May 2019.The seawater has dissolved oxygen(DO) concentration of 3.01-6.68 mg/L and the core of the oxygen minimum zones was located between the depths of 650 m and 1 750 m.The bacterial alpha-diversity showed unimodal pattern with the decreasing DO with depths and peaked in the upper oxycline(UO) of OMZs.The bacterial community structure of the mixed layer(ML) and the bottom layer clustered and separated from each other,while those of UO and the OMZ core(OM) clustered and overlapped.Overall,bacterial community composition transitioned from being Alphaproteobacteria and Gammaproteobacteria-dominant in ML to being Gammaproteobacteria and Nitrososphaeria/Deltaproteobacteria-dominant in UO and OM,and then changed to being Clostridia and unidentified Actinobacteria-dominant in the bottom layer.Moreover,both bacterial alpha-diversity and the abundant classes fitted varying sectioned functions with DO.The DO solely explained 40.37% of the variation of bacterial community composition among layers(P0.001).The predicted function profiling showed that the water column was predominant by chemoheterotrophy,cyanobacteria,and photoautotrophy in ML,by chemoheterotrophy and nitrate/sulfide cycling in UO and OM,and by chemoheterotrophy and ferme ntation in the bottom layer.Our findings revealed the DO-associated variation in bacterial diversity and community composition,and help to clarify the potential responses of microbes and their involved biogeochemical processes to the expansion and intensification of OMZs.     

Spatiotemporal variation in community structure of marine benthic ciliates in the Yellow Sea during and after macroalgal and giant jellyfish blooms

The annual bloom of the green macroalgal Ulva prolifera from May through July since 2008 and another of giant jellyfish Nemopilema nomurai from June through September have been frequent events in the Yellow Sea. However, the patterns of benthic ciliate communities during and after the blooms are still not known. In combination with analyses of benthic environmental factors, we investigated the distribution and community composition of benthic ciliates in the Yellow Sea in July and November 2011. In July, ciliates had high standing crops and diversity in the northern Yellow Sea, and in the inshore area off the southern Shandong Peninsula, where large numbers of green macroalgae accumulated. In November, the abundance, biomass and diversity of ciliates were high in the sea areas off the Shandong Peninsula and Changjiang estuary, where a large quantity of jellyfish occurred in August. Neither the abundance nor the biomass had significant difference between seasons, or between different compartments of the Yellow Sea. The species number, and both Margalef and Shannon-Wiener indices of ciliates were all significantly higher in November than in July. In both seasons, prostomateans and karyorelicteans consistently constituted the first and second most important ciliate groups in biomass; and carnivorous ciliates constituted the primary feeding type in terms of biomass as well as species richness, followed by bacterivores, algivores and omnivores. Compared with that in June 2007 when no macroalgae occurred, the percentage of small-sized bacterivores (e.g. Metacystis spp., Euplotes spp. and scuticociliates) increased in July 2011. The proportion of carnivorous ciliates increased in November, and this increased dominance of carnivorous ciliates may be a response to the increase in predominance of heterotrophic nanoflagellates, which might in turn be ascribed to an effect of green macroalgal and giant jellyfish blooms in the Yellow Sea.     

Morphology and molecular identification of Ulva forming green tides in Qingdao, China

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.     

Phylogenetic analyses of four species of Ulva and Monostroma grevillei using ITS, rbc L and 18S rDNA sequence data

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.     

Bacterial communities fluctuate in abundance and diversity under simulated oil-contaminated seawater conditions

Marine bacteria have recently been identified as a potent solution for petroleum hydrocarbon degradation in response to hazardous oceanic oil spills. In this study, a mesocosm experiment simulating a petroleum spill event was performed to investigate changes in the abundance, structure, and productivity of bacterial communities in response to oil pollution. Cultured heterotrophic bacteria and total bacteria showed a consistent trend involving an immediate decrease in abundance, followed by a slight increase, and a steady low-level thereafter. However, the changing trend of bacterial productivity based on bacterial biomass and bacterial volume showed the opposite trend. In addition, the density of oil-degrading bacteria increased initially, then subsequently declined. The change in the bacterial community structure at day 0 and day 28 were also analyzed by amplified ribosomal DNA restriction analysis(ARDRA), which indicated that the species diversity of the bacterial community changed greatly after oil pollution. Alphaproteobacteria(40.98%)replaced Epsilonproteobacteria(51.10%) as the most abundant class, and Gammaproteobacteria(38.80%)became the second most dominant class in the whole bacterial community. The bacterial communities in oil-contaminated seawater(32 genera) became much more complex than those found in the natural seawater sample(16 genera). The proportion of petroleum-degrading bacteria in the oil-contaminated seawater also increased. In this study, culture-dependent and culture-independent approaches were combined to elucidate changes in both bacterial productivity and community structure. These findings will contribute to a better understanding of the role that bacteria play in material cycling and degradation in response to oil pollution.     

Characterization of Bioaerosol Bacterial Communities During Hazy and Foggy Weather in Qingdao,China

This study was conducted to evaluate the impact of hazy and foggy weather on the bacterial communities in bioaerosols, for which samples were collected from the Qingdao coastal region on sunny, foggy, and hazy days in January and March 2013. Bacterial community compositions were determined using polymerase chain reaction denaturing gradient gel electrophoresis (PCRDGGE). The bacterial community diversity was found to be high on foggy and hazy days, and the dominant species differed during hazy weather. The Shannon-Wiener index revealed that the bacterial community diversity of coarse particles was higher than that of fine particles in the bioaerosols. The bacterial community diversity of fine particles significantly correlated with relative humidity (RH; r² = 0.986). The cluster analysis results indicated that the bacterial communities on sunny days differed from those on hazy and foggy days. Compared with sunny days, the bacterial communities in the fine particles during hazy weather exhibited greater changes than those in the coarse particles. Most of the sequenced bacteria were found to be closely affiliated with uncultured bacteria. During hazy weather, members of the classes Bacilli and Gammaproteobacteria (Pseudomonas and Acinetobacter) were dominant. The DGGE analysis revealed that Proteobacteria and Firmicutes were the predominant phyla, and their relative percentages to all the measured species changed significantly on hazy days, particularly in the fine particles. Haze and fog had a significant impact on the bacterial communities in bioaerosols, and the bacterial community diversity varied on different hazy days.     

Genetic analysis of floating Enteromorpha prolifera in the Yellow Sea with AFLP marker

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.     

Acute toxicity of live and decomposing green alga Ulva (Enteromorpha) prolifera to abalone Haliotis discus hannai

From 2007 to 2009, large-scale blooms of green algae (the so-called “green tides”) occurred every summer in the Yellow Sea, China. In June 2008, huge amounts of floating green algae accumulated along the coast of Qingdao and led to mass mortality of cultured abalone and sea cucumber. However, the mechanism for the mass mortality of cultured animals remains undetermined. This study examined the toxic effects of Ulva (Enteromorpha) prolifera, the causative species of green tides in the Yellow Sea during the last three years. The acute toxicity of fresh culture medium and decomposing algal effluent of U. prolifera to the cultured abalone Haliotis discus hannai were tested. It was found that both fresh culture medium and decomposing algal effluent had toxic effects to abalone, and decomposing algal effluent was more toxic than fresh culture medium. The acute toxicity of decomposing algal effluent could be attributed to the ammonia and sulfide presented in the effluent, as well as the hypoxia caused by the decomposition process.     

DNA barcoding assessment of green macroalgae in coastal zone around Qingdao,China

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.     

Drivers of coastal bacterioplankton community diversity and structure along a nutrient gradient in the East China Sea

Anthropogenic nutrient discharge poses widespread threats to coastal ecosystems and has increased environmental gradients from coast to sea. Bacterioplankton play crucial roles in coastal biogeochemical cycling, and a variety of factors af fect bacterial community diversity and structure. We used 16 S r RNA gene pyrosequencing to investigate the spatial variation in bacterial community composition(BCC) across five sites on a coast-of fshore gradient in the East China Sea. Overall, bacterial alpha-diversity did not diff er across sites, except that richness and phylogenetic diversity were lower in the of fshore sites, and the highest alpha-diversity was found in the most landward site, with Chl-abeing the main factor. BCCs generally clustered into coastal and of fshore groups. Chl-a explained 12.3% of the variation in BCCs, more than that explained by either the physicochemical(5.7%) or spatial(8.5%) variables. Nutrients(particularly nitrate and phosphate), along with phytoplankton abundance, were more important than other physicochemical factors, co-explaining 20.0% of the variation in BCCs. Additionally, a series of discriminant families(primarily affiliated with Gammaproteobacteria and Alphaproteobacteria), whose relative abundances correlated with Chl-a, DIN, and phosphate concentrations, were identified, implying their potential to indicate phytoplankton blooms and nutrient enrichment in this marine ecosystem. This study provides insight into bacterioplankton response patterns along a coast-of fshore gradient, with phytoplankton abundance increasing in the of fshore sites. Time-series sampling across multiple transects should be performed to determine the seasonal and spatial patterns in bacterial diversity and community structure along this gradient.     

Maixi River estuary to the Baihua Reservoir in the Maotiao River catchment: phytoplankton community and environmental factors

Phytoplankton and environmental variables were measured monthly from July 2009 to August 2011 in the Maixi River from the estuary to Baihua Reservoir in the Maotiao River catchment,southwestern China,to understand phytoplankton community structure and environmental factors.The relationship between phytoplankton community structure and environmental factors including hydrological,meteorological,physical,and chemical variables were explored using multivariate analysis.A total of 81 taxa of phytoplankton were identified,which were mainly composed of chlorophyta,bacillariophyta,and cyanobacteria.The phytoplankton community was dominated by Pseudanabaena limnetica during summer and fall and by Cyclotella meneghiniana during winter and spring.The abundance of phytoplankton ranged from 0.24×104 cells/L to 33.45×106 cells/L,with the minimum occurring during February 2010 and the maximum during July 2009.The phytoplankton community was dominated mainly by cyanobacteria from April to September,and by bacillariophyta and pyrrophyta from October to March.Canonical correspondence analysis showed that temperature,pH values,and orthophosphate were the most important driving factors regulating the composition and dynamics of the phytoplankton community in the estuary.Cyanobacteria and euglenophyta abundance and biomass were affected mainly by temperature and pH values,while most chlorophyta and bacillariophyta were influenced by the concentrations of nutrients.     

Spatiotemporal pattern of bacterioplankton in Donghu Lake

Bacterioplankton play key roles in the biogeochemical cycle and in organic contaminant degradation. The species richness and abundance of bacterial subgroups are generally distinct from each other, and this is attributed to their different functions in aquatic ecosystems. The spatiotemporal variations of eight phylogenetic subgroups （Actinobacteria, Bacteroidetes, Cyanobacteria, Firmicutes, Planctomycetes, alpha-, beta-, and gamma-Proteobacteria） derived from Donghu Lake were investigated using PCR-DGGE fingerprinting, to explore their responses to environmental factors. Results indicate that Actinobacteria and beta-Proteobacteria were the two largest bacterial subgroups detected. These two groups and Bacteroidetes showed clear seasonal patterns in composition of the operational taxonomic unit. Results also suggest that the bacterioplankton subgroups in Donghu Lake were significantly correlated with different environmental factors. In brief, the total nitrogen was one of the major factors regulating all the bacterioplankton except for Actinobacteria. However, total phosphorus, another important eutrophication factor, contributed to the two largest bacterial groups （Actinobacteria and beta-Proteobacteria）, as well as to the Cyanobacteria and Firmicutes. Therefore, the responses of bacterioplankton subgroups to environmental factors were different, and this should be attributed to the differences in the fimctions of different groups.     

Epiphytic bacterial communities on two common submerged macrophytes in Taihu Lake: diversity and host-specificity

Leaves of terrestrial and aquatic plants are home to a wide diversity of bacterial species. However, the diversity and variability of epiphytic bacteria on their submerged plant hosts remains poorly understood. We investigated the diversity and composition of epiphytic bacteria from two common submerged macrophytes: Vallisneria natans and Hydrilla verticillata in Taihu Lake, Jiangsu, China, using methods of terminal restriction fragment length polymorphisms (T-RFLP) and clone library analyses targeted at bacterial 16S rRNA genes. The results show that: (1) the libraries of the two waterweeds contain wide phylogenetic distribution of bacteria, and that the sequences of the two libraries can be separated into 93 OTUs (at 97% similar value); (2) Betaproteobacteria, including Burkholderiales, was the most abundant bacterial group on both plants. Cyanobacteria and Gammaproteobacteria were the second largest groups on V. natans and H. verticillata, respectively. Both clone libraries included some sequences related to those of methanotrophs and nitrogen-fixing bacteria; (3) Cluster analysis of the T-RFLP profiles showed two distinct clusters corresponding to the two plant populations. Both ANOSIM of the T-RFLP data and Libshuff analysis of the two clone libraries indicated a significant difference in epiphytic bacterial communities between the two plants. Therefore, the epiphytic bacterial communities on submerged macrophytes appear to be diverse and host-specific, which may aid in understanding the ecological functions of submerged macrophytes in general.