Ras gene in marine mussels: a molecular level response to petrochemical exposure

Mussels are susceptible to numerous toxicants and are often employed as bioindicators. This study investigated the status of the ras proto-oncogene in Mytilus galloprovincialis following petrochemical exposure. A M. galloprovincialis homologue of the vertebrate ras gene was isolated, showing conserved sequence in regions of functional importance and a high incidence of polymorphic variation. Mutational damage was investigated in mussels chronically exposed to the water-accommodated fraction of #4 fuel-oil (WAF), and in mussels collected along the NW coast of Portugal in sites with different levels of petrochemical contamination. A ras gene point mutation was identified in the codon 35 of one individual exposed to 12.5% WAF. No mutations were detected in mussels from the WAF control or environmental samples. This represents the first report of a ras gene mutation, experimentally-induced by petrochemical exposure, in an invertebrate species.     

青岛近海碳钢锈层中细菌群落结构的多样性

研究海水环境中Q235碳钢锈层中微生物群落的多样性。应用16SrDNA基因文库技术,分别对实海全浸一年的Q235碳钢内外锈层中的微生物进行基因文库的构建和分析。通过克隆测序构建了含有64个OTUs的细菌16SrRNA基因文库,结果表明锈层中微生物种类丰富,主要分布于13个已知的菌门,包括变形菌门;拟杆菌门;硅藻门;酸杆菌门;浮霉菌门;硝化螺旋菌门;厚壁菌门;绿弯菌门;疣微菌门;绿菌门;放线菌门;红藻门以及螺旋体门。内外锈层的优势菌均为变形细菌门,分别占文库序列的46%和53%,其中α-变形菌纲又为门中优势类群,但在内外锈层中还存在明显的系统发育学分歧。     

Mutations in exons of the CYP17-II gene affect sex steroid concentration in male Japanese flounder (Paralichthys olivaceus)

As a specific gene of fish,cytochrome P450c17-Ⅱ(CYP17-Ⅱ) gene plays a key role in the growth,development and reproduction level of fish.In this study,the single-stranded conformational polymorphism(SSCP) technique was used to characterize polymorphisms within the coding region of CYP17-Ⅱ gene in a population of 75 male Japanese flounder(Paralichthys olivaceus).Three single nucleotide polymorphisms(SNPs) were identified in CYP17-Ⅱ gene of Japanese flounder.They were c.G594A(p.G188R),c.G939A and c.G1502A(p.G490D).SNP1(c.G594A),located in exon 4 of CYP17-Ⅱ gene,was significantly associated with gonadosomatic index(GSI).Individuals with genotype GG of SNP1 had significantly lower GSI(P < 0.05) than those with geno-type AA or AG.SNP2(c.G939A) located at the CpG island of CYP17-Ⅱ gene.The mutation changed the methylation of exon 6.Indi-viduals with genotype AA of SNP2 had significantly lower serum testosterone(T) level and hepatosomatic index(HSI) compared to those with genotype GG.The results suggested that SNP2 could influence the reproductive endocrine of male Japanese flounder.How-ever,the SNP3(c.G1502A) located in exon 9 did not affect the four measured reproductive traits.This study showed that CYP17-Ⅱ gene could be a potentially useful candidate gene for the research of genetic breeding and physiological aspects of Japanese flounder.     

蛋白核小球藻(Chlorella pyrenoidosa) Rubisco活化酶基因的克隆与表达分析

采用RACE技术克隆了单细胞绿藻蛋白核小球藻820Rubisco活化酶基因(rca)序列,并用荧光定量PCR方法研究了rca和Rubisco小亚基基因(rbcS)的表达规律。结果获得了1703bp的rcacDNA序列,包括66bp的5’非翻译区、1242bp的开放阅读框和395bp的3’非翻译区。序列比较和分析表明该蛋白核小球藻rca序列与其它绿藻的同源性高达78%—85%;偏好使用以G/C/T结尾的密码子;推测的RCA蛋白等电点和分子量分别为8.44和45.71kDa。荧光定量PCR结果表明随光照时间增加rca与rbcS转录表达逐渐降低;不同盐度处理下rbcS的mRNA量变化不大,而rca在37.5盐度下表达量为25盐度的2.69倍;1.0—3.0mmol/L水杨酸处理rca与rbcS表达均降低。     

北部湾拟穴青蟹(Scylla paramamosain)群体遗传结构及其扩张分析

利用线粒体16SrRNA基因片段分析北部湾拟穴青蟹群体遗传结构及其扩张历史。结果表明,16SrRNA基因片段长度在521—523bp之间,6个拟穴青蟹群体具21种单倍型。群体总体水平的单倍型多样性(h)和核甘酸多样性(π)分别为0.6259和0.001851。群体间遗传距离与地理距离之间无相关性。群体间遗传分化系数(Gst)在0.02008—0.04362之间,基因流(Nm)值在5.48—12.20之间。AMOVA分析表明,群体间遗传变异占0.1%,群体内遗传变异占99.9%。群体间固定指数(Fst)的绝对值在0.00026—0.02977之间,暗示群体间没有产生遗传分化。单倍型邻接树和单倍型中介网络图显示,单倍型没有按采样地形成谱系分支。中性检验和错配分析揭示,北部湾拟穴青蟹群体在历史上经历过群体扩张。群体扩张时间大致在更新世晚期的0.04698—0.01879MY前或0.04520—0.01808MY前。     

导入bar基因获得裙带菜草丁膦抗性配子体

裙带菜(Undaria pinnatifida)配子体经2.5 L气升式光生物反应器快速营养增殖,通过基因枪法将携带草丁膦抗性基因(bar)的载体转入扩增后的配子体,经草丁膦筛选,获得抗性克隆。提取抗性克隆配子体基因组总DNA,经PCR和PCR Southern检测,结果显示60%的抗性克隆可检出bar基因,表明bar基因是裙带菜配子体基因工程有效的筛选标记。     

基于线粒体D-loop 基因探讨花鳗鲡的群体遗传多样性及其种群进化历史

通过测定花鳗鲡(Anguillia marmorata)海南群体(HN)和菲律宾群体(PH)共19尾个体的线粒体D-loop基因的核苷酸序列(约1017 bp),分析了花鳗鲡的种群遗传结构。结果表明:A、T、G、C 4种核苷酸的平均含量分别为39.8%、28.3%、12.4%、19.4%,A+T含量(68.1%)明显高于G+C含量(31.8%)。所测序列中存在73个变异位点,共有18个单倍型。其中海南群体的单倍型多样度(Hd)、核苷酸多态性(Pi)、平均核苷酸差异数(k)分别为0.982、0.21577和219.655,而菲律宾群体的单倍型多样度、核苷酸多态性、平均核苷酸差异数分别为1.000、0.26728和271.821,两个群体之间平均遗传距离(P)为0.3203。结果表明2个群体遗传变异较大,菲律宾群体的遗传多样性较海南群体更加丰富。通过构建NJ分子系统树表明2个群体的亲缘关系较近。利用中性检验Tajima’s D(海南群体D=1.35345,P>0.01;菲律宾群体D=0.79220,P>0.01)和Fs(海南群体Fs=3.759;菲律宾群体Fs=2.231)探讨其种群历史,表明花鳗鲡群体进化过程种群数量较为稳定。