Development of an in situ loop-mediated isothermal amplification technique for chromosomal localization of DNA sequences     

孟庆磊  王师  张玲玲  黄晓婷  包振民 《中国海洋湖沼学报》2013,31(1):128-133

In situ loop-mediated isothermal amplification (in situ LAMP) combines in situ hybridization and loop-mediated isothermal amplification (LAMP) techniques for chromosomal localization of DNA sequences. In situ LAMP is a method that is generally more specific and sensitive than conventional techniques such as fluorescence in situ hybridization (FISH), primed in situ labeling (PRINS), and cycling primed in situ labeling (C-PRINS). Here, we describe the development and application of in situ LAMP to identify the chromosomal localization of DNA sequences. To benchmark this technique, we successfully applied this technique to localize the major ribosomal RNA gene on the chromosomes of the Zhikong scallop (Chlamys farreri).  相似文献   

三种壳口颜色脉红螺（Rapanavenosa）形态学和线粒体16SrRNA与COI基因片段差异比较分析     

班绍君  薛东秀  张涛  潘洋  王海艳 《海洋与湖沼》2012,43(6)

脉红螺壳口通常有3种颜色：“黑白条纹螺”、“中间螺”和“橙色螺”。本论文对3种壳口颜色的脉红螺分别进行了形态学和mt16SrRNA、COI基因片段序列比较分析。形态学分析结果表明,除厣宽／壳高、厣高／壳高、厣高／体螺层、厣高／壳口长、厣高／壳口宽、厣高／厣宽的形态学特征指数和出肉率不具有显著性差异外,其它形态特征和生物学相关变量具有显著差异,且对湿重的影响效果不同。“黑白条纹螺”壳口长和壳口宽对湿重影响最大,而“中间螺”和“橙色螺”各形态特征对湿重均无显著的直接影响。根据各形态特征均值建立的居群形态聚类图显示,“中间螺”和“橙色螺”形态差异较小,二者与“黑白条纹螺”的形态差异较大。然而基于mt16SrRNA和COI基因片段序列分析结果显示,三种壳口颜色的脉红螺无遗传分化,为同一种。  相似文献   

基于线粒体16S rRNA 基因研究5 个栉江珧野生群体的遗传多样性   总被引：1，自引：0，他引：1  

严加坤  杨爱国  周丽青  吴彪  白临建  刘志鸿 《海洋科学》2013,37(2):36-42

采用通用引物对山东长岛、文登、日照、广东湛江和海南三亚等5个地理群体栉江珧(Atrina pectinta)16S rRNA序列进行扩增、测序分析,得到59条441bp的核苷酸序列.其中T、C、A、G和A+T的平均含量分别为29.91%、17.41%、25.74%、26.94%及55.65%, AT含量高于GC含量,共检测到了9个单倍型和26个核苷酸多态位点.群体多样性分析表明,文登群体具有较高的遗传多样性水平.AMOVA 分析表明,五群体间总遗传分化系数Fst =0.5007(P<0.001),群体间遗传分化略大于群体内、群体间存在较高的遗传分化.基于群体间遗传距离构建NJ和UPGMA分子进化树,5个地理群体的栉江珧聚为两个支,长岛、文登、日照群体聚为一支,海南和湛江群体聚为另一支.  相似文献   

荣成天鹅湖大叶藻(Zostera marina)间2种小型腹足类的分类学研究     

刘静  管洪在  张素萍  郭小芳  杨学伶  王海艳 《海洋与湖沼》2013,44(1):226-234

小型腹足类生物在我国种类繁多,但其相关研究在国内基本属于空白.本文对荣成天鹅湖大叶藻间的2种小型腹足类生物进行了形态学及分子生物学(16S和28S rRNA基因片段)研究.结果表明,根据形态学所鉴定的短剑螺属未定种Machaeroplax sp.,应为畦螺属未定种Lirularia sp.;分子生物学研究结果支持形态学对刺绣翼螺Alaba picta的鉴定.同时,与A.picta有一定形态学差异的其它4种相近形态的小型腹足类,在基因序列及遗传距离上与刺绣翼螺非常接近,亦应为刺绣翼螺Alaba picta.  相似文献   

长牡蛎外套膜组织着色区和无色区的微生物多样性比较分析     

谢超伊  宋宏策  董美云  张钰煊  高凯利  魏磊  王晓通 《海洋通报》2024,(2):188-197

典型的开管式软体动物长牡蛎（Crassostrea gigas）具有开闭壳的行为习惯,易使大量海水成分进入体内。因此,海水中的微生物极易附着于长牡蛎的软体部表面。本研究利用16S rRNA对长牡蛎外套膜着色区和无色区以及其生活区域海水进行了微生物多样性比较分析。结果显示：长牡蛎外套膜表面的微生物多样性与丰富度均高于海水。在门水平上,变形菌门（Proteobacteria）为长牡蛎外套膜着色区与无色区的共同优势菌类,而海水中优势菌类为拟杆菌门（Bacteroidetes）。在属水平上,主要发现了弧菌（Vibrio）、假交替单孢菌（Pseudoalteromonas）和海洋单胞菌（Marinomonas）在内的28个菌属。此外,线性判别分析（Linear discriminant analysis effect size,LeFSe）显示,外套膜着色区与无色区的微生物标志物存在差异。着色区的主要标志物为螺旋体科未分类属（norank＿f＿Spirochaetaceae）,而无色区的主要标志物为弧菌（Vibrio）。综上,长牡蛎外套膜组织表面与海水之间的微生物组成存在较大差异。尽管外套膜着色...  相似文献   

基于三种分子标记的外伶仃岛石珊瑚系统进化关系研究     

肖宝华  廖宝林  杨小东  谢子强 《海洋科学》2017,41(9):1-8

以广东省珠海市外伶仃岛地区常见的4属5种石珊瑚为研究对象,对线粒体COI、16S rRNA和ITS三种基因片段数据进行分析,并计算了种间遗传距离。序列分析结果表明5种石珊瑚的COI、16S rRNA基因片段碱基AT含量大于GC含量,而ITS序列碱基AT含量小于GC含量;5种石珊瑚种间的三种基因平均遗传距离分别为0.043、0.134和0.763,所得遗传距离结果存在一定差异。将三种基因串联起来并利用邻位连接法(NJ)、最大似然法(ML)和最小进化法(ME)分别构建了分子系统树。在系统发育树中,科一级的系统进化关系与形态学分类一致,但蜂巢珊瑚科内的分子系统分类结果与形态学分类阐述的遗传进化关系略有差别,说明石珊瑚传统分类可能受骨骼可塑性的影响。研究结果为外伶仃岛石珊瑚的亲缘关系研究提供相关基础数据,并为该地区石珊瑚资源保护奠定理论基础。  相似文献   

黑鲷(Acanthopagrus schlegelii)肠道细菌群落对饲料中添加不同羽毛粉的响应   总被引：2，自引：0，他引：2  

邓惠文  刘峰  李桂豪  包卫洋  邹松保  郑鹏飞  龚骏  孙汝江 《海洋与湖沼》2019,50(6):1309-1317

羽毛粉是一种潜在的饲料蛋白源。在水产动物饲料中添加适量的羽毛粉,不仅可以降低饲料成本,而且有助于禽类废弃物的再利用。在动物消化、降解、吸收羽毛粉过程中,动物肠道中的微生物可能发挥着重要作用;添加羽毛粉的饲料对鱼类肠道微生物多样性与群落结构的影响目前还不清楚。基于细菌16SrRNA基因高通量测序,作者对比研究了以添加羽毛粉(包括水解、酸解、膨化处理或生羽毛粉实验组,添加量为5%)和不添加羽毛粉的饲料(即对照组,包括鱼粉和商业饲料两组)喂养黑鲷12周后,黑鲷肠道细菌多样性与群落结构的变化情况。统计分析结果表明:添加羽毛粉的实验组中肠道细菌的丰富度、辛普森指数和香农指数显著低于对照组(P0.05);不含酸解羽毛粉组的实验组,其细菌群落结构与对照组差异显著(P0.05)。总体来看,黑鲷肠道细菌主要由厚壁菌门(57.3%)和变形菌门(36.3%)组成;实验组细菌群落中拟杆菌纲、梭菌纲、δ-变形菌纲、草酸杆菌科和青枯菌属的相对丰度显著低于对照组。不同的羽毛粉处理组之间也呈现出群落结构上的差异:与水解羽毛粉组相比,生羽毛粉组的变形菌门、γ-变形菌门、海洋螺菌目、盐单胞菌科和盐单胞菌属的相对丰度显著下降;与鱼粉组相比,生羽毛粉组中芽孢杆菌目在群落中的占比显著上升,而拟杆菌门、草酸杆菌科和青枯菌属的占比则显著下降,而且生羽毛粉组中厚壁菌门与变形菌门相对丰度的比值(7.42)明显高于鱼粉组(0.97)和水解羽毛粉组(0.74)且均接近显著(P=0.057)。与鱼粉组相比,水解羽毛粉组中紫单胞菌科和回线属的相对丰度显著下降,而盐单胞菌属的相对丰度显著上升。  相似文献   

Bacterial community succession in response to dissolved organic matter released from live jellyfish     

HAO Wenjin  WICHELS Antje  FUCHS Bernhardt  TANG Xuexi  GERDTS Gunnar 《海洋湖沼学报（英文）》2019,(4):1229-1244

Jellyfish blooms have increased worldwide, and the outbreaks of jellyfish population not only affect the food web structures via voracious predation but also play an important role in the dynamics of nutrients and oxygen in planktonic food webs. However, it remains unclear whether specific carbon compounds released through jellyfish metabolic processes have the potential to shape bacterial community composition. Therefore, in this study, we aimed to investigate the compositional succession of the bacterioplankton community in response to the dissolved organic matter (DOM) released by the live Scyphomedusae Cyanea lamarckii and Chrysaora hysoscella collected from Helgoland Roads of the North Sea. The bacterial community was significantly stimulated by the DOM released form live jellyfish and different dominant phylotypes were observed for these two Scyphomedusae species. Furthermore, the bacterial community structures in the different DOM sources, jellyfish-incubated media, Kabeltonne seawater, and artificial seawater (DOM-free) were significantly different, as revealed by automated ribosomal intergenic spacer analysis fingerprints. Catalyzed reporter deposition fluorescence in situ hybridization (CARD-FISH) revealed a rapid species-specific shift in bacterial community composition. Gammaproteobacteria dominated the community instead of the Bacteroidetes community for C. lamarckii, whereas Gammaproteobacteria and Bacteroidetes dominated the community for C. hysoscella. The significant differences in the bacterial community composition and succession indicate that the components of the DOM released by jellyfish might differ with jellyfish species.  相似文献   

Molecular analyses of bacterioplankton communities with highly abundant Vibrio clades: a case study in Bohai Sea coastal waters     

WANG Shuping  ZHANG Yuan  HE Jia  JIA Xiaobo  LIN Jianing  LI Meng  WANG Qinglin 《海洋湖沼学报（英文）》2019,(5):1638-1648

We investigated the bacterioplankton abundance, community composition and the associated Vibrio clades of natural seawater in Bohai Sea coastal waters. Seawater samples (10 L in triplicate) were collected at 0.5, 3, and 5 m depths near the coastal aquaculture zone of the Bohai Sea on May 12, 2016. Real-time PCR and 16S rRNA gene amplicon high-throughput sequencing methods were employed by which 485 operational taxonomic units (OTUs) at a 97% sequence similarity level were generated. Gammaproteobacteria, Alphaproteobacteria, and Bacteroidetes were the most abundant groups, accounting for 49.5%, 23.5%, and 18.9% of the total assemblage, respectively. Obvious variations in Pseudoalteromonas, Vibrio, and Octadecabacter , which were the most abundant genera, could be observed among diff erent samples. Notably, the results of Vibrio -specifi c real-time PCR indicated that Vibrio had extremely high 16S rRNA gene copy numbers. The 16S rRNA gene sequencing results across all the samples also indicated that they occupied a large proportion of the total assemblage. Both the alpha diversity and major bacterioplankton group Pseudoalteromonas had significant correlations with the concentration of PO4^3-. Overall, studies on bacterioplankton communities with highly abundant Vibrio clades can provide interesting insight into the microbial function and health assessment of the Bohai Sea coastal ecosystem.  相似文献   

中国海膨胀巴夫藻(定鞭藻门,巴夫藻纲)的首次纪录     

张晓东  杨双  孙军  乔延龙  王静  刘海娇 《海洋学报(英文版)》2018,37(10):28-32

2015年5月,我们从天津近海分离出一株黄绿色具鞭毛的单细胞藻。这珠藻细胞形态不规则,从圆形到细长型都有出现。绝大多数细胞具有很强的运动能力,一些细胞也可以形成静止的群体。结合形态学、超显微结构观察以及分子生物学分析,我们将这株藻鉴定为定鞭藻门,巴夫藻纲的膨胀巴夫藻。在此之前,中国近海只有绿色巴夫藻的记录,绿色巴夫藻对于水产养殖起到了很重要的作用。巴夫藻纲的其他物种尚无中国海记录。这是膨胀巴夫藻在中国海的首次记录。  相似文献