波吉卵囊藻胞外滤液对铜绿微囊藻转录水平的影响

【目的】探索波吉卵囊藻胞外滤液对铜绿微囊藻的抑制机理。【方法】采用Illumina平台,对使用BG11培养基(对照组)和波吉卵囊藻胞外滤液(实验组)培养的铜绿微囊藻(Microcystis aeruginosa)进行转录组测序,筛选差异表达基因并进行通路富集分析,并对部分差异表达基因进行qRT-PCR验证。【结果】与对照组相比,波吉卵囊藻胞外滤液处理后的铜绿微囊藻组中筛选1483个表达差异显著的基因,其中上调表达基因788个,下调表达基因695个。GO功能分析将差异基因划分为35个子类别,包括催化活性、结合、代谢过程、细胞过程、膜、膜部分等。KEGG通路分析中,所有差异基因聚集在108个通路,其中84个差异基因注释在代谢功能类别中,而氧化磷酸化通路富集最显著。氧化磷酸化通路中,相关基因表达以上调为主(26个DEGs,20个上调,6个下调)。qRT-PCR验证结果与转录组测序结果的表达变化趋势一致。【结论】波吉卵囊藻滤液引起铜绿微囊藻氧化磷酸化途径上基因表达上调,ATP合成效率提高。     

Cultural characteristics of chromium resistant unicellular cyanobacteria isolated from local environment in Pakistan

Many unicellular cyanobacteria were isolated from different places： fields, ponds, polluted water, and soils from Muredkey and Kasur tannery areas, near Lahore, Pakistan. Different media like BG 11 medium, Bold Basal medium, Chu＇s # 10 medium and Gotham＇s medium, in standard forms and with slight variations of ingredients, and different pH, temperature and light regimes were checked for the optimum growth of the isolates. The isolation pro- cedure was repeated with different concentrations of chromium to select the resistant strains. These selected strains grew on chromium of the range 100-200 μg/ml in BG 11 medium. Cyanobacteria were maintained in solid and liquid media with/without shaking. Cyanobacterial strains were collected from natural habitats that were accompanied by a diversified group of organisms including bacteria, protozoan, and rotifers etc. In order to eliminate these agents termed as contaminants, we used several methods including phenol treatment, use of antibiotic and careful manual picking of unicellular cyanobacteria. Resistance of these strains against different heavy metals （ZnSO4, MnSO4, NiSO4, COCl2, Pb（NO3）3, CuSO4, HgCl2, AgNO3 and CdCl2） and antibiotics （erythromycin, streptomycin, kanamycin, chloramphenicol, neomycin） was evaluated. Optimum temperature was 30℃ with variable pH for the reduction of Cr^6＋ in to Cr^3＋ in majority of strains.     

Genetic transformation of <Emphasis Type="Italic">Nannochloropsis oculata</Emphasis> with a bacterial phleomycin resistance gene as dominant selective marker

The gene ble from Streptoalloteichus hindustanus is widely used as a selective antibiotic marker. It can control the phleomycin resistance, and significantly increase the tolerance of hosts to zeocin. The unicellular marine microalga Nannochloropsis oculata is extremely sensitive to zeocin. We selected ble as the selective marker for the genetic transformation of N. oculata. After the algal cells at a density of 2×10⁷ cells mL^?1 was digested with 4% hemicellulase and 2% driselase for 1 h, the protoplasts accounted for 90% of the total. The ble was placed at the downstream of promoter HSP70A-RUBS2 isolated from Chlamydomonas reinhardtii, yielding a recombinant expression construct pMS188. The construct was transferred into the protoplasts through electroporation (1 kV, 15 μS). The transformed protoplasts were cultured in fresh f/2 liquid medium, and selected on solid f/2 medium supplemented with 500 ng mL^?1 zeocin. The PCR result proved that ble existed in the transformants. Three transformants had been cultured for at least 5 generations without losing ble. Southern blotting analysis showed that the ble has been integrated into the genome of N. oculata. The ble will serve as a new dominant selective marker in genetic engineering N. oculata.     

Abundance, biomass and composition of spring ice algal and phytoplankton communities of the Laptev Sea （Arctic）

Abundance,biomass and composition of the ice algal and phytoplank-ton communities were investigated in the southeastern Laptev Sea in spring 1999.Diatoms dominated the algal communities and pennate diatoms dominated the dia-tom population.12 dominant algal species occurred within sea ice and underlyingwater column,including Fragilariopsis oceanica,F.cylindrus,Nitzschiafrigida,N.promare,Achnanthes taeniata,Nitzschia neofrigida,Naviculapelagica,N.vanhoef fenii,N.septentrionalis,Melosira arctica,Clindrothecaclosterium and Pyrarnimonas sp.The algal abundance of bottom 10 cm sea icevaried between 14.6 and 1562.2×10~4 ceils l~(-1)with an average of 639.0×10~4cells l~(-1),and the algal biomass ranged from 7.89 to 2093.5μg C l~(-1)with an av-erage of 886.9μg C l~(-1),which were generally one order of magnitude higherthan those of sub-bottom ice and two orders of magnitude higher than those ofunderlying surface water.The integrated algal abundance and biomass of lower-most 20 cm ice column were averagely 7.7 and 12.2 times as those of upper 20 mwater column,respectively,suggesting that the ice algae might play an importantrole in maintaining the coastal marine ecosystem before the thawing of sea ice.Icealgae influenced the phytoplankton community of the underlying water column.However,the“seeding”of ice algae for phytoplankton bloom was negligible be-cause of the iow phytoplankton biomass within the underlying water column.     

Transcriptome Analysis of Crassostrea sikamea (♀) × Crassostrea gigas (♂) Hybrids Under and After Thermal Stress

Crossbreeding is an effective approach to manage the genetic decline in aquaculture.One-way hybrids of Crassostrea sikamea(♀)and Crassostrea gigas(♂)have advantages in growth traits and adaptation to high temperature.Here,we used high-throughput sequencing to analyze the molecular processes in the hybrids under and after thermal stress.The hybrids were cultured in the seawater with an increasing temperature from 25℃to 40℃during 10 hours,which is regarded as the thermal stress stage.Then the temperature decreased from 40℃to 25℃within 2 h,which is regarded as the recovery stage.In this study,1293 significant diffe-rentially expressed genes(DEGs)were obtained under thermal stress,of which 576 were upregulated and 717 were downregulated,and 740 significant differentially expressed genes(DEGs)were obtained in the recovery stage,of which the number of upregulated and downregulated genes was 409 and 331,respectively.The antigen processing and presentation,NOD-like,and NF-kappa B path-ways were significantly enriched during the thermal stress stage.The MAPK and PPAR signaling pathways were significantly enrich-ed during the recovery stage.The HSP70,HSP90,and CANX genes were strongly and rapidly upregulated in the control/thermal stress groups but were slightly less upregulated in the thermal stress/recovery group.These results indicate that the innate immune system or nonspecific immunity was deployed to protect interior tissues from thermal stress.In addition,85%of the mutual DEGs were involved in bidirectional regulation(up/down or down/up)when the oysters were removed from the thermal stress to recover.This study provides preliminary insight into the molecular response of C.sikamea(♀)and C.gigas(♂)hybrids to thermal stress and provides a basis for future studies on temperature-adaptation and the possible expansion of hybrid breeding.