Genetic diversity and population connectivity of the sea urchin Tripneustes gratilla along the South African coast

The collector sea urchin Tripneustes gratilla has been identified as a species with potential for aquaculture production in South Africa. The species’ roe is considered a culinary delicacy in Asia and Europe. However, T. gratilla remains genetically uncharacterised in South Africa. Therefore, the purpose of this study was to provide baseline genetic information consisting of estimates of genetic diversity and population stratification, which may aid in future sustainable use of this urchin resource. A total of 22 species-specific microsatellite markers were used for the genetic characterisation of T. gratilla samples from along the South African coast, at Haga Haga, Coffee Bay, Hibberdene, Ballito Bay and Sodwana Bay. A moderate level of genetic diversity was observed, with an average number of alleles of 7.89 and an average effective number of alleles of 6.57, as well as an average observed heterozygosity of 0.55. Population differentiation tests suggest that the geographically representative samples form part of a single, large interbreeding population, with a global F_st estimate of 0.02 (p > 0.05). This finding is likely explained by high levels of gene flow between these locations caused by extensive larval dispersal during the planktonic larval stage. The panmixia observed within these natural populations of T. gratilla indicate that they could be managed as a single genetic stock.     

基于形态学和16S rRNA基因序列的轮螺科面盘幼虫物种鉴定

轮螺科(Architectonicidae)是一类成体螺壳右旋而面盘幼虫螺壳左旋的腹足类,其面盘幼虫在国内一直被误定为强卷螺属(Agadina)种类,归在螔螺科(Limacinidae)中。本文基于线粒体16SrRNA基因序列和胚壳的形态特征,对轮螺科面盘幼虫进行了物种鉴定。结果表明,根据胚壳壳顶和脐孔的形态特征,面盘幼虫可大致分为2种明显不同的形态类型:形态类型I的个体壳扁,壳顶凹陷,脐孔形状规则,圆而深;形态类型II的个体壳顶突出,脐孔形状不规则,浅或深,肛区龙骨有或无。GMYC(GeneralizedMixedYuleCoalescent)和ABGD(AutomaticBarcodingGapDiscovery)分析显示,轮螺科面盘幼虫16S rRNA基因的50种单倍型形成19个分子可操作分类单元(molecular operationaltaxonomic units,MOTUs),同一MOTU的幼虫具有相同的胚壳形态。其中,11个MOTUs中的幼虫属于形态类型I,8个MOTUs中的幼虫属于形态类型II。此外,不同MOTUs的面盘幼虫,其软体部黑色幼体器官(Black Larval Organ)的位置和大小也不同。根据研究结果推测,轮螺面盘幼虫的形态具有种或属特异性。轮螺科面盘幼虫单倍型形成的MOTU数量明显多于国内目前已记录的物种数,其种类多样性可能被低估。基于16SrRNA序列能直接鉴定到种的仅Psilaxisradiatus和配景轮螺(Architectonica perspectiva)2种。本文也从分子水平订正了国内长期以来的分类错误。     

北江大刺鳅(Mastacembelus armatus)的核型分析及线粒体Cyt b基因和D-loop的遗传多样性

采用胸腔注射植物凝集素(PHA)、秋水仙素,以大刺鳅(Mastacembelus armatus)头肾组织为材料,低渗处理,空气干燥法进行染色体标本制备,分析其染色体核型。结果显示:大刺鳅的二倍体染色体数目为2n=48,其核型公式为2n=14m+34t, NF=62。对24尾大刺鳅线粒体Cyt b基因和D-loop序列进行测序分析,结果表明:(1)大刺鳅Cytb基因、D-loop序列长度分别为1438bp、895bp,两个基因的单倍型多样性分别为0.656和0.541,核苷酸多样性分别为0.00539和0.00817,平均核苷酸差异数分别为5.938和6.805,说明该种群的遗传多样性丰富。(2)两个基因的每个个体的遗传距离分别为0.000—0.018和0.000—0.026,说明该种群个体间的遗传距离小。(3)中性检验的Tajima’sD检验及Fu's Fs检验的结果说明该种群没有发生过扩张。结果说明北江大刺鳅的遗传多样性水平较高,可用于人工繁育。     

Isolation and Characterization of Pathogenic Listonella anguillarum of Diseased Haft-Smooth Tongue Sole （Cynoglossus semilaevis Günther）

Studies were conducted to determine the cause of the acute mortality of half-smooth tongue sole Cynoglossus semilaevis Günther juveniles in a fish farm in Jimo, Shandong Province, China, in June 2006. Gross signs of the diseased tongue sole included several petechiae and ecchymoses on the body and fin necrosis and hemorrhagic lesion at the base of the fin. Bacteria were isolated from kidney, liver and hemorrhagic lesions of the diseased tongue sole. Among 14 strains, SJ060621 was proved to be highly virulent to juvenile tongue sole with LD50 value of 〈1.0×10^5 colony forming units （CFU）mL^-1, while the remaining 13 were avirulent. Among the 16 antibiotics tested, SJ060621 was sensitive to gentamicin and nitrofurantoin. It was identified as Listonella anguillantm with conventional plate and tube tests in combination with API 20E analysis. 16S rRNA gene and partial HSP60 gene sequenceing analysis revealed that the strain was highly homologous with L. anguillarum. Examination of the infected musculature by electron microscopy indicated numerous bacteria and lots of macrophages containing phagocytosed bacteria. Histopathological investigations revealed severe necrotic degenerative changes in the infected organs. Indirect immunofluorescence assay （IFA） was employed to detect the location of occurrence of bacteria, and bacteria were found in aggregations in the inflammatory areas in musculature.     

Isolation and characterization of pathogenic <Emphasis Type="Italic">Listonella anguillarum</Emphasis> of diseased half-smooth tongue sole (<Emphasis Type="Italic">Cynoglossus semilaevis</Emphasis> Günther)

Studies were conducted to determine the cause of the acute mortality of half-smooth tongue sole Cynoglossus semilaevis Günther juveniles in a fish farm in Jimo, Shandong Province, China, in June 2006. Gross signs of the diseased tongue sole included several petechiae and ecchymoses on the body and fin necrosis and hemorrhagic lesion at the base of the fin. Bacteria were isolated from kidney, liver and hemorrhagic lesions of the diseased tongue sole. Among14 strains, SJ060621 was proved to be highly virulent to juvenile tongue sole with LD₅₀ value of <1.0×10⁵ colony forming units (CFU) mL⁻¹, while the remaining 13 were avirulent. Among the 16 antibiotics tested, SJ060621 was sensitive to gentamicin and nitrofurantoin. It was identified as Listonella anguillarum with conventional plate and tube tests in combination with API 20E analysis. 16S rRNA gene and partial HSP60 gene sequenceing analysis revealed that the strain was highly homologous with L. anguillarum. Examination of the infected musculature by electron microscopy indicated numerous bacteria and lots of macrophages containing phagocytosed bacteria. Histopathological investigations revealed severe necrotic degenerative changes in the infected organs. Indirect immunofluorescence assay (IFA) was employed to detect the location of occurrence of bacteria, and bacteria were found in aggregations in the inflammatory areas in musculature.     

Cloning and characterization of proliferating cell nuclear antigen gene of Alexandrium catenella (Dinoflagellate) with respect to cell growth

Harmful algal blooms (HABs) have been affecting negatively the shellfish and aquaculture industries around the world. Though a lot of efforts have been made to disclose the changes of environmental factors involved and their effects on the HABs events, the molecular mechanism of this process remains unclear. To address this problem, proliferating cell nuclear antigen gene (pcna) was isolated and characterized from Alexandrium catenella. It showed high homology to those of other dinoflagellates (89% and 91% homology to Pfiesteria piscicid and Pyrocystis lunula, respectively), and also 42%-43% homology to those of plant and animals. The expression level of pcna revealed by quantitative real time PCR was the lowest at the late lagging cell growth phase, increased to the highest at the late exponential phase, and then decreased at the stationary phase. Though the cell growth rate was also changing, no positive correlation between pcna expression level and cell growth rate was displayed throughout the whole cell growth stages (r²=0.024 6). However, the pcna expression level had the similar trend with the change of cell growth rate throughout the whole growing process, e.g., from increasing at the earlier cell growth stage to decreasing at the following stages, though slightly lagging to the latter.     

High gene flow promotes the genetic homogeneity of the fish goby Pomatoschistus marmoratus (Risso, 1810) from Mar Menor coastal lagoon and adjacent marine waters (Spain)

The extreme environmental variability of coastal lagoons suggests that physical and ecological factors could contribute to the genetic divergence among populations occurring in lagoon and open‐coast environments. In this study we analysed the genetic variability of lagoon and marine samples of the sand goby, Pomatoschistus marmoratus (Risso, 1810) (Pisces: Gobiidae), on the SW Spain coast. A fragment of mitochondrial DNA control region (570 bp) was sequenced for 196 individuals collected in five localities: Lo Pagan, Los Urrutias and Playa Honda (Mar Menor coastal lagoon), and Veneziola and Mazarrón (Mediterranean Sea). The total haplotype diversity was h = 0.9424 ± 0.0229, and the total nucleotide diversity was π = 0.0108 ± 0.0058. Among‐sample genetic differentiation was not significant and small‐scale patterns in the distribution of haplotypes were not apparent. Gene flow and dispersal‐related life history traits may account for low genetic structure at a small spatial scale. The high genetic diversity found in P. marmoratus increases its potential to adapt to changing conditions of the Mar Menor coastal lagoon.     

On four closely related hypotrichous ciliates (Protozoa, Ciliophora, Spirotrichea): molecular characters, interspecific relationships and phylogeny defined with multigene sequence information

In order to clarify the phylogeny and relationships of the most confused hypotrichous ciliates, Holosticha-complex, four closely related holostichids (five populations), Holosticha bradburyae, H. diademata, Anteholosticha sp., and A. manca, were compared and analyzed using ITS2 secondary structures, ITS1-5.8S-ITS2 region and SSrRNA gene sequences. The ITS1-5.8S-ITS2 region sequences of these four species were first sequenced, and they shared sequence identities ranging from 68.0% to 90.1%, while two populations of Anteholosticha sp. differed in three nucleotides (sequence identity 99.8%). There were several minor differences among ITS2 secondary structures of these species, while two populations of Anteholosticha sp. had the identical secondary structure. Phylogenetic trees inferred from the ITS1-5.8S-ITS2 region sequences of stichotrichs using multiple algorithms (Neighbor-Joining, Maximum Parsimony and Bayesian) revealed similar topologies. The results show that:(1) Holosticha bradburyae and H. diademata firmly clustered together with strong bootstrap supports, forming a sister clade with Anteholosticha sp., (2) Anteholosticha appeared to be a paraphyletic assemblage, in which the morphotype A. manca was more closely related to Diaxonella trimarginata than to its congener Anteholosticha sp. Phylogenetic analyses based on the SSrRNA gene and the combined sequences of SSrRNA gene and ITS1-5.8S-ITS2 region revealed the similar relationships between Holosticha and Anteholosticha, nevertheless their positions within the subclass Stichotrichia differed from each other inferred from different genes.     

太湖光合自养真核超微藻遗传多样性初探

采用真核藻类叶绿体16S rRNA的引物PLA491F、OXY1313R,通过PCR、构建基因克隆库的方法初步调查了太湖梅梁湾和东太湖光合自养真核超微藻的遗传多样性.结果表明,太湖自养型真核超微藻主要为隐藻(Cryptophyta),其次为硅藻(Bacillariophyta)、金藻(Chrysophyta)和定鞭藻(Haptophyta).该结果有助于我们更全面地了解太湖浮游藻类的群落结构及生态功能.     

2022年夏季胶州湾致灾物种多棘海盘车的分子解析

多棘海盘车(Asterias amurensis)是海星纲的一个致灾物种,广泛分布于包括我国黄海、渤海海域、韩国、日本和俄罗斯海域在内的北太平洋海域,并作为入侵种分布于澳大利亚的塔斯马尼亚海域。多棘海盘车在多个海域发生过暴发性增殖导致的高密度、大规模聚集,对海洋生态系统和水产养殖造成严重的负面影响。自2006年以来在我国山东青岛海域多次发生大规模多棘海盘车聚集,对贝类养殖造成重大经济损失。日本、俄罗斯、澳大利亚等海域的多棘海盘车遗传多样性研究取得了显著进展,然而,迄今为止鲜有针对我国海域致灾物种多棘海盘车的分子生物学分析,阻碍了我国海域种群与全球种群间遗传进化关系的比较分析。研究重点分析了于2022年7月在胶州湾发生多棘海盘车暴发时采集的12个样本,系统组装了其线粒体基因组、核糖体基因簇以及多种通用分子标记。基于线粒体基因组和核糖体基因簇的系统发育分析表明多棘海盘车具有较高的种内遗传多样性。基于线粒体基因组的蛋白编码基因进行分化时间估算,多棘海盘车的种内遗传分化可能发生在1.7～4.2 Ma,表明多棘海盘车的扩布可能较早发生。与线粒体基因组相比,多棘海盘车核糖体基因簇序列在种内极为保守...