氮磷营养盐添加对二甲基巯基丙酸内盐合成与降解细菌及其功能基因的影响

【目的】二甲基巯基丙酸内盐(dimethylsulfoniopropionate,DMSP)是海洋中主要的有机硫化物之一,是海洋细菌硫的主要来源,海洋细菌将其分解成"冷室气体"二甲基硫(dimethylsulfide,DMS),对调节全球气候变化和驱动地球硫循环有重要作用。本研究通过中国东海水体的现场围隔实验模拟海水富营养化对DMSP、DMS产量以及DMSP合成基因(dsyB和mmtN)和降解基因(dddP和dmdA)及相关功能细菌的影响。【方法】通过流式细胞仪计数92个围隔海水样品中微微型浮游生物的数量,采用Illumina MiSeq测序技术对海水样品中细菌的16S rRNA基因进行高通量测序,利用荧光定量PCR技术定量测定16S rRNA基因、DMSP合成及降解基因的丰度。【结果】研究发现,同时添加硝酸盐(6.00μmol/L)和磷酸盐(0.375μmol/L)能促进叶绿素a、DMSP、DMS的浓度上升。对于DMSP合成基因,只加磷酸盐能促进dsyB及Phaeobacter等相应物种的富集,虽然同时添加硝酸盐和磷酸盐使dsyB富集,但相对只加磷酸盐却不利于dsyB积累;同时添加硝酸盐和磷酸盐也抑制Alteromonas的生长,进而抑制了mmtN的富集。对于DMSP降解基因,同时加入硝酸盐和磷酸盐促进了dddP及Thalassococcus、Thalassobius、Loktanella和Shimia等物种的富集,却抑制了SAR11、Sulfitobacter等的富集,从而导致dmdA无法被富集。【结论】氮限制能更好地促进DMSP合成基因的表达,从而迫使细菌增加DMSP的合成以应对氮营养条件不足的生存环境,并进而提高DMSP脱甲基化的比例为细菌提供更多能量;而在硝酸盐和磷酸盐充足情况下,细菌相对减少DMSP的合成且更倾向于裂解DMSP产生DMS来降低硫同化的比例。本研究结果强调了海水富营养化对细菌合成与降解DMSP过程的影响。     

Diversity of the cyanobacterial microflora of the northern part of James Ross Island,NW Weddell Sea,Antarctica

The diversity and ecological distribution of cyanobacteria in the northern, deglaciated part of James Ross Island were studied during the Antarctic summer season 2005–2006. Seventy-five cyanobacterial morphotypes were observed in various habitats of this area. The identified cyanobacterial taxa belong to the characteristic and dominant types of coastal Antarctica, and majority of them appeared connected to special habitats and formed distinct populations and ecologically delimited communities. The results are compared and discussed with respect to phenotypically characterised cyanobacterial microflora of maritime Antarctica and to recent molecular analyses of cyanobacterial strains from different Antarctic regions. The existence of a specificity and characteristic composition of Antarctic cyanobacterial communities was demonstrated.     

Functional evaluation of a nitrogenase-like protochlorophyllide reductase encoded by the chloroplast DNA of Physcomitrella patens in the cyanobacterium Leptolyngbya boryana

Dark-operative protochlorophyllide (Pchlide) oxidoreductase (DPOR) is a nitrogenase-like enzyme consisting of the two components, L-protein (a ChlL dimer) and NB-protein (a ChlN-ChlB heterotetramer), to catalyze Pchlide reduction in Chl biosynthesis. While nitrogenase is distributed only among certain prokaryotes, the probable structural genes for DPOR are encoded by chloroplast DNA in lower plants. Here we show functional evaluation of DPOR encoded by chloroplast DNA in a moss Physcomitrella patens by the complementation analysis of the cyanobacterium Leptolyngbya boryana and the heterologous reconstitution of the moss L-protein and the cyanobacterial NB-protein. Two shuttle vectors to overexpress chlL and chlN-chlB from P. patens were introduced into the cyanobacterial chlL- and chlB-lacking mutants, respectively. Both transformants restored the ability to perform Chl biosynthesis in the dark, indicating that the chloroplast-encoded DPOR components form an active complex with the cyanobacterial components. The L-protein of P. patens was purified from the cyanobacterial transformant, and DPOR activity was reconstituted in a heterologous combination with the cyanobacterial NB-protein. The specific activity of the L-protein from P. patens was determined to be 118 nmol min(-1) mg (-1), which is even higher than that of the cyanobacterial L-protein (76 nmol min(-1) mg (-1)). Upon exposure to air, the activity of the L-protein from P. patens decayed with a half-life of 30 s, which was eight times faster than that of the cyanobacterial L-protein (240 s). These results suggested that the chloroplast-encoded L-protein functions as efficiently as the cyanobacterial L-protein but is more oxygen labile than the cyanobacterial L-protein.     

蓝细菌神经毒素研究进展

蓝细菌飞速繁殖形成水华能够造成严重的危害,其中某些蓝细菌由于能够产生毒素而受到广泛关注,但是我国对于产神经毒素的蓝细菌一直研究较少。我们对蓝细菌神经毒素近年来的研究进展进行简要综述。     

Engineering cyanobacteria to generate high-value products

Although many microorganisms have been used for the bioindustrial generation of valuable metabolites, the productive potential of cyanobacterial species has remained largely unexplored. Cyanobacteria possess several advantages as organisms for bioindustrial processes, including simple input requirements, tolerance of marginal agricultural environments, rapid genetics, and carbon-neutral applications that could be leveraged to address global climate change concerns. Here, we review recent research involving the engineering of cyanobacterial species for the production of valuable bioindustrial compounds, including natural cyanobacterial products (e.g. sugars and isoprene), biofuels (e.g. alcohols, alkanes and hydrogen), and other commodity chemicals. Biological and economic obstacles to scaled cyanobacterial production are highlighted, and methods for increasing cyanobacterial production efficiencies are discussed.     

Efficacy of New Inexpensive Cyanobacterial Biofertilizer Including its Shelf-life

Summary Four cyanobacterial inoculants all significantly increased grain and straw yield of rice either alone or in combination with chemical fertilizer. A saving of 25 kg N ha⁻¹ can be attained through cyanobacterial fertilization. Tobacco waste-based cyanobacterial biofertilizer was best in performance. Cyanobacterial acetylene reducing activity in vivo varied from 144 to 255 μmol C₂H₄ m⁻² h⁻¹ in different treatments, being highest for tobacco-based cyanobacterial biofertilizer integrated with 50% chemical N. The nutrient balance for total N, available N, total P and available P was found positive in biofertilizer- and chemical fertilizer-treated plots. The total and available K showed negative balance in all the treatments. The shelf-life of cyanobacterial biofertilizer can be augmented by selecting translucent packing material, dry mixing and paddy straw as a carrier. Dry mixing and a mixing ratio of 50:50 (carrier:cyanobacteria) gave better inoculum loading and shelf-life. Decrease in cyanobacterial population was least in dried cyanobacterial flacks, indicating a possibility of developing cyanobacterial biofertilizer without carrier mixing at the time of production.     

Cyanobacterial peptides - nature's own combinatorial biosynthesis

Cyanobacterial secondary metabolites have attracted increasing scientific interest due to bioactivity of many compounds in various test systems. Among the known structures, oligopeptides are often found with many congeners sharing conserved substructures, while being highly variable in others. A major part of known oligopeptides are of non-ribosomal origin and can be grouped into classes with conserved structural properties. Thus, the overall structural diversity of cyanobacterial oligopeptides only seemingly suggests an equally high diversity of biosynthetic pathways and respective genes. For each class of peptides, some of which have been found in all major branches of the cyanobacterial evolutionary tree, homologous synthetases and genes can be inferred. This implies that non-ribosomal peptide synthetase genes are a very ancient part of the cyanobacterial genome and presumably have evolved by recombination and duplication events to reach the present structural diversity of cyanobacterial oligopeptides. In addition, peptide synthetases would appear to be an essential part of the cyanobacterial evolution and physiology. The present review presents an overview of the biosynthesis of cyanobacterial peptides and corresponding gene clusters, the structural diversity of structural types and structural variations within peptide classes, and implications for the evolution and plasticity of biosynthetic genes and the potential function of cyanobacterial peptides.     

The carboxyl-terminal region of the spinach PsaD subunit contains information for its specific assembly into plant thylakoids

The assembly of the multi-subunit membrane-protein Photosystem I (PS I) complex involves incorporation of peripheral proteins into the complex. Here we studied assembly of the PsaD subunit of the cyanobacterial and plant PS I into the thylakoid membranes. We generated partial and chimeric psaD genes from which labeled proteins were synthesized in vitro. Assembly of these proteins into the cyanobacterial or plant thylakoids was assayed. The deletion of leader sequence and N-terminal extension of spinach prePsaD did not inhibit its assembly into spinach or cyanobacterial thylakoids. Addition of these sequences to the cyanobacterial PsaD did not enable it to assemble into plant thylakoids. Moreover, these additions significantly decreased the ability of the chimeric proteins to assemble into cyanobacterial thylakoids. In contrast, when the carboxyl-terminal half of cyanobacterial PsaD was replaced by the corresponding region of the spinach PsaD, the chimeric protein could assemble into both spinach and cyanobacterial thylakoids. Therefore, information in the carboxyl-terminal region of spinach PsaD is crucial for its assembly into plant thylakoids.Abbreviation prePsaD precursor of the PsaD subunit of PS I     

An early origin of plastids within the cyanobacterial divergence is suggested by evolutionary trees based on complete 16S rRNA sequences

It is generally accepted that the plastids arose from a cyanobacterial ancestor, but the exact phylogenetic relationships between cyanobacteria and plastids are still controversial. Most studies based on partial 16S rRNA sequences suggested a relatively late origin of plastids within the cyanobacterial divergence. In order to clarify the exact relationship and divergence order of cyanobacteria and plastids, we studied their phylogeny on the basis of nearly complete 16S rRNA gene sequences. The data set comprised 15 strains of cyanobacteria from different morphological groups, 1 prochlorophyte, and plastids belonging to 8 species of plants and 12 species of diverse algae. This set included three cyanobacterial sequences determined in this study. This is the most comprehensive set of complete cyanobacterial and plastidial 16S rRNA sequences used so far. Phylogenetic trees were constructed using neighbor joining and maximum parsimony, and the reliability of the tree topologies was tested by different methods. Our results suggest an early origin of plastids within the cyanobacterial divergence, preceded only by the divergence of two cyanobacterial genera, Gloeobacter and Pseudanabaena.      

Convergent photophysiology and prokaryotic assemblage structure in epilithic cyanobacterial tufts and algal turf communities

As global change spurs shifts in benthic community composition on coral reefs globally, a better understanding of the defining taxonomic and functional features that differentiate proliferating benthic taxa is needed to predict functional trajectories of reef degradation better. This is especially critical for algal groups, which feature dramatically on changing reefs. Limited attention has been given to characterizing the features that differentiate tufting epilithic cyanobacterial communities from ubiquitous turf algal assemblages. Here, we integrated an in situ assessment of photosynthetic yield with metabarcoding and shotgun metagenomic sequencing to explore photophysiology and prokaryotic assemblage structure within epilithic tufting benthic cyanobacterial communities and epilithic algal turf communities. Significant differences were not detected in the average quantum yield. However, variability in yield was significantly higher in cyanobacterial tufts. Neither prokaryotic assemblage diversity nor structure significantly differed between these functional groups. The sampled cyanobacterial tufts, predominantly built by Okeania sp., were co-dominated by members of the Proteobacteria, Firmicutes, and Bacteroidota, as were turf algal communities. Few detected ASVs were significantly differentially abundant between functional groups and consisted exclusively of taxa belonging to the phyla Proteobacteria and Firmicutes. Assessment of the distribution of recovered cyanobacterial amplicons demonstrated that alongside sample-specific cyanobacterial diversification, the dominant cyanobacterial members were conserved across tufting cyanobacterial and turf algal communities. Overall, these data suggest a convergence in taxonomic identity and mean photosynthetic potential between tufting epilithic cyanobacterial communities and algal turf communities, with numerous implications for consumer-resource dynamics on future reefs and trajectories of reef functional ecology.     

Use of a rapid bioluminescence assay for detecting cyanobacterial microcystin toxicity

The recent rise in the awareness of the occurrence of toxic cyanobacterial blooms in aquatic environments, with associated human health problems and animal deaths, has increased the need for rapid, reliable and sensitive methods of determining cyanobacterial toxicity. A luminescent bacterial toxicity test was assessed as a complement to the established mouse bioassay. Seventeen samples including pure cyanobacterial microcystin-LR hepatotoxin, laboratory isolates and natural blooms of cyanobacteria were tested and toxicity data compared with mouse LD50 values. Microcystin-LR and all five microcystin-containing cyanobacterial samples, hepatotoxic by mouse test gave EC50 values of less than 0.46 mg/ml in bioluminescence-based Microtox assays. Of 11 samples non-toxic by mouse bioassay, only two gave an EC50 of less than 0.98 mg/ml by bioluminescence assay. It is suggested that the Microtox bioluminescence assay may prove useful in the preliminary screening of cyanobacterial blooms for microcystin-based toxicity.     

Anopheles albimanus (Diptera: Culicidae) and cyanobacteria: an example of larval habitat selection

Northern Belize has extensive herbaceous wetlands. Those dominated by sparse emergent macrophytes, rushes (Eleocharis spp.) and sawgrass (Cladium jamaicense Crantz), often develop floating mats of cyanobacteria (blue-green algae). These mats provide suitable habitat for larvae of the malaria transmitting mosquito Anopheles albimanus Wiedemann. Presence/absence of A. albimanus larvae and cyanobacterial mats was assessed in marshes located throughout northern Belize. Of the 21 marshes examined during the 1993 wet and 1994 dry seasons, cyanobacterial mats were found in 11 and A. albimanus larvae were detected in 9 of these 11 marshes. No A. albimanus larvae were found in marshes without cyanobacterial mats. Mosquito larvae were collected along two 1,000 m long transects in both the wet season (August 1993) and the dry season (March 1994) to delineate larval distribution in marshes with cyanobacterial mats. A. albimanus larval densities in cyanobacterial mats were relatively high in both seasons: 2.8 and 2.3 larvae per dip in the wet and dry seasons, respectively, in Chan Chen marsh; and 0.8 and 1.02 larvae per dip in Buena Vista marsh. Numbers of larvae per dip did not significantly change with increasing distance from houses/pastures or margins of the marsh. A field experiment showed a strong preference of ovipositing A. albimanus for cyanobacterial mats. Higher temperatures and higher CO2 emissions from cyanobacterial mats are possible ovipositional cues.     

Epilithic and Cryptoendolithic Cyanobacteria of Clarens Sandstone Cliffs in the Golden Gate Highlands National Park,South Africa

A cryptoendolithic cyanobacterial community is described from Clarens sandstone in a montane ecosystem of the north-eastern Orange Free State, South Africa. The community is dominated by a species of the unicellular cyanobacterial genus Chroococcidiopsis. Chlorophyll_a content of the colonised sandstone ranged between 44.3 and 51.8mg m^?2. The establishment and species composition of an artificially created tintenstrich cyanobacterial community is documented. The possible contribution of cryptoendolithic cyanobacterial communities in the biological formation of hollows and caves in Clarens sandstone cliffs is discussed.     

Transgenic plants with cyanobacterial genes

Over the years, cyanobacteria have been regarded as ideal model systems for studying fundamental biochemical processes like oxygenic photosynthesis and carbon and nitrogen assimilation. Additionally, they have been used as human foods, sources for vitamins, proteins, fine chemicals, and bioactive compounds. Aiming to increase plant productivity as well as nutritional values, cyanobacterial genes involved in carbon metabolism, fatty acid biosynthesis, and pigment biosynthesis have been intensively exploited as alternatives to homologous gene sources. In this short review, transgenic plants with cyanobacterial genes generated over the last two decades are examined, and the future prospects for transgenic crops using cyanobacterial genes obtained from functional genomics studies of numerous cyanobacterial genomes information are discussed.     

Function of the IsiA pigment–protein complex in vivo

Photosynthesis Research - The acclimation of cyanobacterial photosynthetic apparatus to iron deficiency is crucial for their performance under limiting conditions. In many cyanobacterial species,...     

Comparative and evolutionary aspects of cyanobacteria and plant plastid division study

The review summarizes the current understanding of cyanobacterial cell division mechanisms in comparison with those of eukaryotic plastids. It also sheds light on the present knowledge of the nature of evolutionary transformations of the cyanobacterial cell division apparatus that could have occurred during the establishment of modern plastid division complex. Peculiar properties of cyanobacterial cell division process are discussed as well as the features of primary and secondary plastid replication.     

Preliminary analysis to estimate the spatial distribution of benefits of P load reduction: Identifying the spatial influence of phosphorus loading from the Maumee River (USA) in western Lake Erie

Since the early 2000s, Lake Erie has been experiencing annual cyanobacterial blooms that often cover large portions of the western basin and even reach into the central basin. These blooms have affected several ecosystem services provided by Lake Erie to surrounding communities (notably drinking water quality). Several modeling efforts have identified the springtime total bioavailable phosphorus (TBP) load as a major driver of maximum cyanobacterial biomass in western Lake Erie, and on this basis, international water management bodies have set a phosphorus (P) reduction goal. This P reduction goal is intended to reduce maximum cyanobacterial biomass, but there has been very limited effort to identify the specific locations within the western basin of Lake Erie that will likely experience the most benefits. Here, we used pixel‐specific linear regression to identify where annual variation in spring TBP loads is most strongly associated with cyanobacterial abundance, as inferred from satellite imagery. Using this approach, we find that annual TBP loads are most strongly associated with cyanobacterial abundance in the central and southern areas of the western basin. At the location of the Toledo water intake, the association between TBP load and cyanobacterial abundance is moderate, and in Maumee Bay (near Toledo, Ohio), the association between TBP and cyanobacterial abundance is no better than a null model. Both of these locations are important for the delivery of specific ecosystem services, but this analysis indicates that P load reductions would not be expected to substantially improve maximum annual cyanobacterial abundance in these locations. These results are preliminary in the sense that only a limited set of models were tested in this analysis, but these results illustrate the importance of identifying whether the spatial distribution of management benefits (in this case P load reduction) matches the spatial distribution of management goals (reducing the effects of cyanobacteria on important ecosystem services).     

Phylogenetic analysis of symbiotic and free-living cyanobacterial cultures using DNA amplification fingerprinting

DNA amplification fingerprinting (DAF) of 17 cyanobacterial isolates belonging to symbiotic and free-living forms of 6 different genera was done. The dendrogram analysis of 17 cyanobacterial cultures revealed three major clusters. All Westiellopsis cultures formed the first major cluster and their nucleotide relatedness ranged between 71-93%. In the second major cluster, the symbiotic cyanobacterium Anabaena sp dominated and all Anabaena cultures showed 60-75% similarity with each other. Nostoc muscorum is related to Anabaena variabilis by 85% and formed the third major cluster. The dendrogram analysis of cyanobacterial isolates clearly revealed that free-living cyanobacterial cultures were closely related with each other and were diverse from the symbiotic forms.     

The possibility of using cyanobacterial bloom materials as a medium for white rot fungi

Aims: The present study was conducted to evaluate the possibility of using cyanobacterial bloom materials as a medium for white rot fungi and the capability of white rot fungi, Trichaptum abietinum 1302BG and Lopharia spadicea to biodegrade dried cyanobacterial bloom material taken from Taihu Lake. Methods and Results: The results showed T. abietinum 1302BG and L. spadicea could use the cyanobacterial bloom materials taken from Taihu Lake for growth to measure the mycelial plaque and dry‐weight mycelial pellicles of fungi. The removal rate of dried cyanobacterial bloom materials incubated with white rot fungi is approximately 100%. Conclusions: The cyanobacterial bloom material can be used as a glucose substitute in white rot fungi medium. The white rot fungi, T. abietinum 1302BG and L. spadicea, can also directly decrease the biomass of cyanobacterial bloom material taken from Taihu Lake. Significance and Impact of the Study: Cyanobacterial bloom thrives in eutrophic fresh waters all over the world. Micro‐organisms, particularly fungi, have attracted attention as possible agents for the degradation of phytoplankton species. Dealing with cyanobacterial bloom material as a medium for fungi instead of directly discharging them as organic fertilizers is a new, safe and environmentally friendly approach.     

Seasonal dynamics of chemotypes in a freshwater phytoplankton community – A metabolomic approach

Cyanobacteria are known to produce a huge variety of secondary metabolites. Many of these metabolites are toxic to zooplankton, fish, birds and mammals. Therefore, the toxicity of cyanobacterial blooms is strongly dependent on the cyanobacterial strain composition. These strains produce distinct bouquets of secondary metabolites, and thus constitute different chemotypes. Some of the cyanobacterial metabolites are potent inhibitors of gut proteases of the filter-feeder Daphnia. Here, we investigate the seasonal dynamics of secondary metabolites in a phytoplankton community from a hypertrophic pond, making use of a new metabolomic approach. Using liquid chromatography coupled with high-resolution mass spectrometry (LCMS), we obtained mass spectra of phytoplankton samples taken on different dates throughout the summer season. By applying multivariate statistics, we combined these data with the protease inhibition capacity of the same samples. This led to the identification of metabolites with cyanobacterial origin and as well of distinct cyanobacterial chemotypes being dominant on different dates. The protease inhibition capacity varied strongly with season, and only one out of 73 known cyanobacterial protease inhibitors could be confirmed in the natural samples. Instead, several so far unknown, putative protease inhibitors were detected. In conclusion, the creation of time series of mass spectral data of a natural phytoplankton community proved to be useful for elucidating seasonal chemotype succession in a cyanobacterial community. Additionally, correlating mass spectral data with a biological assay provides a promising tool for facilitating the search for new harmful metabolites prior to structure elucidation.