Institute of Hydrobiology, Chinese Academy Of Sciences

    人工试验湖泊浮游藻类群落的生态学研究

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    为了对转基因(CAgcGH)鲤的生态风险评估提供参考资料,于2002年构建人工试验湖泊。研究分析了该人工湖泊浮游藻类群落的结构特征、季节动态、年际变化及其与水体各环境因子的关系。2006年至2010年间,每季度采样,共鉴定出浮游藻类7门47属66种,其中绿藻种类最多。双向指示种分析(TWINSPAN)和除趋势对应分析(DCA)结果显示采样点数据可分为春夏秋冬4组,说明该群落季节性明显。冬季群落结构简单,多样性最低,主要由小环藻(Cyclotella sp.)和分歧锥囊藻(Dinobryon divergens)组成;春季,小环藻、针杆藻(Synedra sp.)、颗粒直链藻(Melosira granulata)等几种硅藻占优势;夏季群落结构复杂,占优势的是银灰平裂藻(Merismopedia glauca)和螺旋鞘丝藻(Lyngbya contarta),多样性最高;秋季没有明显占优势的种类。5年间,群落细胞密度上升了33.1%,平均值为(1.43±0.75)×106cells/L;硅藻在群落中所占比例从48.2%下降至16.2%,而蓝藻从9.3%上升至42.2%。典范对应分析(CCA)的结果显示对浮游藻类影响最大的环境因子是温度和溶氧,总氮浓度和总磷浓度的影响也是不可忽视的,而pH在试验中对浮游藻类群落结构的影响有限。不同藻类在CCA排序图上有不同的分布格局,一些硅藻主要分布在中低温采样点,蓝藻集中分布在高温的采样点,鼓藻主要出现在高透明度和高总磷浓度的采样点,金藻主要分布在高溶氧浓度和低温的采样点

    Sequencing and analysis of the complete genome of Rana grylio virus (RGV)

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    Infection with Rana grylio virus (RGV), an iridovirus isolated in China in 1995, resulted in a high mortality rate in frogs. The complete genome sequence of RGV was determined and analyzed. The genomic DNA was 105,791 bp long, with 106 open reading frames (ORFs). Dot plot analysis showed that the gene order of RGV shared colinearity with three completely sequenced ranaviruses. A phylogenetic tree was constructed based on concatenated sequences of iridovirus 26 core-gene-encoded proteins, and the result showed high bootstrap support for RGV being a member of the genus Ranavirus and that iridoviruses of other genera also clustered closely. A microRNA (miRNA) prediction revealed that RGV could encode 18 mature miRNAs, many of which were located near genes associated with virus replication. Thirty-three repeated sequences were found in the RGV genome. These results provide insight into the genetic nature of RGV and are useful for laboratory diagnosis for vertebrate iridoviruses.Infection with Rana grylio virus (RGV), an iridovirus isolated in China in 1995, resulted in a high mortality rate in frogs. The complete genome sequence of RGV was determined and analyzed. The genomic DNA was 105,791 bp long, with 106 open reading frames (ORFs). Dot plot analysis showed that the gene order of RGV shared colinearity with three completely sequenced ranaviruses. A phylogenetic tree was constructed based on concatenated sequences of iridovirus 26 core-gene-encoded proteins, and the result showed high bootstrap support for RGV being a member of the genus Ranavirus and that iridoviruses of other genera also clustered closely. A microRNA (miRNA) prediction revealed that RGV could encode 18 mature miRNAs, many of which were located near genes associated with virus replication. Thirty-three repeated sequences were found in the RGV genome. These results provide insight into the genetic nature of RGV and are useful for laboratory diagnosis for vertebrate iridoviruses

    Taxonomic and phylogenetic evaluation of Limnothrix strains (Oscillatoriales, Cyanobacteria) by adding Limnothrix planktonica strains isolated from central China

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    Six Limnothrix strains, isolated for the first time from a shallow eutrophic lake in central China, were taxonomically and phylogenetically evaluated by investigating their polyphasic characteristics, including morphological features, cellular ultrastructures, and 16S rRNA gene sequences. All the six strains were morphologically similar, and their trichomes were in average 1.7 mu m wide and cells 4.0 mu m long, and having small gas vesicles within cells, and therefore identified as Limnothrix planctonica (Woloszynska) Meffert. Cellular ultrastructures of them showed that peripheral thylakoids with 3-5 parallel layers were parietally distributed in the cells. The phylogenetic results based on the 16S rRNA gene sequences showed that all the Limnothrix strains, including the six in this study and those from the Genbank, formed two distinct clusters. The similarity in 16S rDNA sequences between these two clusters was lower than 90%, indicating that these Limnothrix strains belong to different genera. This is the first report on the morphology and phylogeny of L. planctonica strains, providing the new information on taxonomy of the genus Limnothrix.Six Limnothrix strains, isolated for the first time from a shallow eutrophic lake in central China, were taxonomically and phylogenetically evaluated by investigating their polyphasic characteristics, including morphological features, cellular ultrastructures, and 16S rRNA gene sequences. All the six strains were morphologically similar, and their trichomes were in average 1.7 mu m wide and cells 4.0 mu m long, and having small gas vesicles within cells, and therefore identified as Limnothrix planctonica (Woloszynska) Meffert. Cellular ultrastructures of them showed that peripheral thylakoids with 3-5 parallel layers were parietally distributed in the cells. The phylogenetic results based on the 16S rRNA gene sequences showed that all the Limnothrix strains, including the six in this study and those from the Genbank, formed two distinct clusters. The similarity in 16S rDNA sequences between these two clusters was lower than 90%, indicating that these Limnothrix strains belong to different genera. This is the first report on the morphology and phylogeny of L. planctonica strains, providing the new information on taxonomy of the genus Limnothrix

    水产动物重要经济性状的分子基础及其遗传改良

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    中国水产养殖的成就已被认为是中国对当今世界发展的一个重大贡献.近20年来,基因组学和其他遗传技术的进步显著推动着水产动物分子生物学和遗传育种学基础研究的发展.进入21世纪后,一些主要水产动物的全基因组测序已经完成或接近完成.本文综述了包括基因组技术、体细胞核移植和干细胞技术在内的水产遗传改良技术的一些重要突破性进展,概述了包括生殖、性别、生长、抗病、耐寒和耐低氧这些重要经济性状的分子基础,列出了一系列与这些经济性状相关的候选基因.最后,介绍了几个特别是中国水产动物遗传改良在水产养殖中的应用事例,强调了未来的发展方向

    异源四倍体银鲫外周血和精巢的组织学特征

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    通过比较D系三倍体银鲫(Carassius auratus gibelio Bloch)与异源四倍体银鲫,我们发现异源四倍体的外周血与精巢组织跟三倍体银鲫存在明显差异。HE染色结果表明,异源四倍体银鲫外周血红细胞有明显的分裂倾向。利用流式细胞术对D系三倍体银鲫与异源四倍体银鲫外周血的DNA直方图进行比较,结果表明异源四倍体外周血的DNA直方图有两个主峰。此外,我们观察到异源四倍体银鲫精巢的三种类型,其中Ⅰ型精巢可以产生正常精子,Ⅱ型可观察到精小囊结构,但不能产生精子,Ⅲ型精巢未发育出精小囊结构。进一步用银鲫Vasa抗体对精巢切片进行组织免疫荧光共聚焦显微分析,结果表明,Ⅰ型精巢的生殖细胞完成了减数分裂,能观察到精原细胞、初级精母细胞、次级精母细胞,以及大量位于精小管中间的精子细胞和精子;而Ⅱ型精巢的生殖细胞不能完成第二次减数分裂,精小囊中存在大量的初级和次级精母细胞,没有精子细胞产生。研究丰富了对异源四倍体银鲫生物学性状的认识

    Hydrogen peroxide induces apoptotic-like cell death in Microcystis aeruginosa (Chroococcales, Cyanobacteria) in a dose-dependent manner

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    We investigated the capability of Microcystis aeruginosa to cause apoptosis by pursuing morphological, molecular and physiological characteristics after exposure to H2O2. Microcystis proliferation was only weakly affected after exposure to 150 mu M H2O2 but cell numbers decreased dramatically after exposures of 250 and 325 mu M H2O2. Cells exposed to 250 and 325 mu M H2O2 were examined using transmission electron microscopy, and they exhibited membrane deformation and partial disintegration of thylakoids. Correspondingly, fluorescence imaging of DNA by Hoechst 33342 staining revealed the condensation of nucleoid chromatin. Moreover, cellular injury was concomitant with dramatic decreases in photosynthetic efficiency (ratio of variable fluorescence to maximum fluorescence [Fv/Fm], maximum electron transport rate [ETRmax]) and elevated caspase-3-like activity after exposure of 250 and 325 mu M H2O2. Terminal deoxynucleotidyl transferase Deoxyuridine 5-triphosphate nick end labelling (TUNEL) positive staining appeared in cells exposed to 250 mu M and 325 mu M H2O2, and the percentage staining increased with increasing H2O2 concentration. These data suggested that M. aeruginosa exposed to H2O2 underwent an apoptotic event. Additionally, cells exposed to H2O2 had increased cytoplasmic vacuolation and nontypical DNA laddering. Increased caspase-3-like activity was not inhibited in the presence of the synthetic caspase inhibitor carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone. Therefore, H2O2 induced apoptotic-like cell death in a dose-dependent manner. Taken together, our results provided a novel mechanism for explaining cyanobacterial bloom dynamics in response to environmental stress. The results also contributed to the understanding of the origin and evolution of programmed cell death.We investigated the capability of Microcystis aeruginosa to cause apoptosis by pursuing morphological, molecular and physiological characteristics after exposure to H2O2. Microcystis proliferation was only weakly affected after exposure to 150 mu M H2O2 but cell numbers decreased dramatically after exposures of 250 and 325 mu M H2O2. Cells exposed to 250 and 325 mu M H2O2 were examined using transmission electron microscopy, and they exhibited membrane deformation and partial disintegration of thylakoids. Correspondingly, fluorescence imaging of DNA by Hoechst 33342 staining revealed the condensation of nucleoid chromatin. Moreover, cellular injury was concomitant with dramatic decreases in photosynthetic efficiency (ratio of variable fluorescence to maximum fluorescence [Fv/Fm], maximum electron transport rate [ETRmax]) and elevated caspase-3-like activity after exposure of 250 and 325 mu M H2O2. Terminal deoxynucleotidyl transferase Deoxyuridine 5-triphosphate nick end labelling (TUNEL) positive staining appeared in cells exposed to 250 mu M and 325 mu M H2O2, and the percentage staining increased with increasing H2O2 concentration. These data suggested that M. aeruginosa exposed to H2O2 underwent an apoptotic event. Additionally, cells exposed to H2O2 had increased cytoplasmic vacuolation and nontypical DNA laddering. Increased caspase-3-like activity was not inhibited in the presence of the synthetic caspase inhibitor carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone. Therefore, H2O2 induced apoptotic-like cell death in a dose-dependent manner. Taken together, our results provided a novel mechanism for explaining cyanobacterial bloom dynamics in response to environmental stress. The results also contributed to the understanding of the origin and evolution of programmed cell death

    Mitochondrial divergence suggests unexpected high species diversity in the opsariichthine fishes (Teleostei: Cyprinidae) and the revalidation of Opsariichthys macrolepis

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    Opsariichthine (sensu Oceanologi Et Limnologia Sinica, 1982, 13, 293-298) is a cyprinid group consisting of five genera and endemic to East Asia. Previous studies suggested that there may be many possible cryptic species in this group, but this has not been confirmed. In this study, using mitochondrial cyt b sequences on 1,388 samples and 739 haplotypes, we showed very high species diversity within this group. The results showed that phylogenetic relationships of the opsariichthine group were as ([Nipponocypris-Parazacco-Candidia] + [Zacco + Opsariichthys]), and there were multiple deep lineages within several species, flagging putative cryptic species. When a 3% genetic distance was used as a threshold for species delimitation, 35 haplogroups were found, nine haplogroups in Candidia-Parazacco-Nipponocypris group, six haplogroups in the Zacco group, and 20 haplogroups in the Opsariichthys group. We consider all of them to be putative until determination of distinct species based on the tree topology, geographic distributions, or a combination of both. In addition, two kinds of species delimitation tools, ABGD and PTP, were applied to construct molecular operational taxonomic units (MOTUs). The ABGD method revealed nine MOTUs in Candidia-Parazacco-Nipponocypris group, two MOTUs in the Zacco group, and 17 MOTUs in the Opsariichthys group. And the PTP method revealed 10 MOTUs in Candidia-Parazacco-Nipponocypris group, 10 MOTUs in the Zacco group, and 29 MOTUs in the Opsariichthys group. Therefore, there should be more species in the opsariichthine group than presently described. Based on the molecular data and morphological characteristics, we proposed Opsariichthys macrolepis as a valid species and described its morphological diagnostic characters.</p

    Mga Modulates Bmpr1a Activity by Antagonizing Bs69 in Zebrafish

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    MAX giant associated protein (MGA) is a dual transcriptional factor containing both T-box and bHLHzip DNA binding domains. In vitro studies have shown that MGA functions as a transcriptional repressor or activator to regulate transcription of promotors containing either E-box or T-box binding sites. BS69 (ZMYND11), a multidomain-containing (i.e., PHD, BROMO, PWWP, and MYND) protein, has been shown to selectively recognizes histone variant H3.3 lysine 36 trimethylation (H3.3K36me3), modulates RNA Polymerase II elongation, and functions as RNA splicing regulator. Mutations in MGA or BS69 have been linked to multiple cancers or neural developmental disorders. Here, by TALEN and CRISPR/Cas9-mediated loss of gene function assays, we show that zebrafish Mga and Bs69 are required to maintain proper Bmp signaling during early embryogenesis. We found that Mga protein localized in the cytoplasm modulates Bmpr1a activity by physical association with Zmynd11/Bs69. The Mynd domain of Bs69 specifically binds the kinase domain of Bmpr1a and interferes with its phosphorylation and activation of Smad1/5/8. Mga acts to antagonize Bs69 and facilitate the Bmp signaling pathway by disrupting the Bs69-Bmpr1a association. Functionally, Bmp signaling under control of Mga and Bs69 is required for properly specifying the ventral tailfin cell fate.</p

    Rana grylio virus 43R encodes an envelope protein involved in virus entry

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    Rana grylio virus (RGV), a member of genus Ranavirus in the family Iridoviridae, is a viral pathogen infecting aquatic animal. RGV 43R has homologues only in Ranavirus and contains a transmembrane (TM) domain, but its role in RGV infection is unknown. In this study, 43R was determined to be associated with virion membrane. The transcripts encoding 43R and the protein itself appeared late in RGV-infected EPC cells and its expression was blocked by viral DNA replication inhibitor, indicating that 43R is a late expressed protein. Subcellular localization showed that 43R-EGFP fusion protein distributed in cytoplasm of EPC cells and that TM domain is essential for its distribution in cytoplasm. 43R-EGFP fusion protein colocalized with viral factories in RGV-infected cells. A recombinant RGV deleting 43R (43R-RGV) was constructed by homologous recombination to investigate its role in virus infection. Compared with wild type RGV, the ability of 43R-RGV to induce the cytopathic effect and its virus titers were significantly reduced. Furthermore, it is revealed that 43R deletion significantly inhibited viral entry but did not influence viral DNA replication by measuring and comparing the DNA levels of RGV and 43R-RGV in the infected cells at the early stage of infection. RGV neutralization with anti-43R serum reduced the virus titer. Therefore, these data showed that RGV 43R is a late gene that encodes an envelope protein involved in RGV entry.</p

    Effects of Salt Concentrations and Nitrogen and Phosphorus Starvations on Neutral Lipid Contents in the Green Microalga Dunaliella tertiolecta

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    Dunaliella tertiolecta, a halotolerant alga, can accumulate large amounts of neutral lipid, which makes it a potential biodiesel feedstock. In this study, neutral lipids of D. tertiolecta induced by different salinities or N or P starvation were analyzed by thin-layer chromatography (TLC), flow cytometry (FCM), and confocal laser scanning microscopy (CLSM). High salinities or N or P starvation resulted in a decrease in cell growth and chlorophyll contents of D. tertiolecta. Neutral lipid contents increased markedly after 3-7 days of N starvation or at low NaCl concentrations (0.5-2.0 M). N starvation had a more dramatic effect on the neutral lipid contents of D. tertiolecta than P starvation. Four putative ME isozymes in different conditions can be detected by using isozyme electrophoresis. Two alternative acetyl-CoA producers, ACL and ACS genes, were up-regulated under low salinities and N starvation. It was suggested that low salinities and N starvation are considered efficient ways to stimulate lipid accumulation in D. tertiolecta.</p
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