花染色体组型分析及细胞核DNA含量的测定

采用PHA、秋水仙碱腹腔或背部肌肉注射,活体培养法,取花肾细胞制成悬浮液,用空气干燥法制片。经Giemsa染色,镜检细胞分裂相,统计结果为2n=50,核型公式为:2n=20m+12sm+10st+8t,NF=82。同时,取花肌肉和肝脏为材料,以鸡血细胞为DNA标准(2.30pg/2c),使用EPICS-XL型流式细胞仪测定了花细胞DNA含量,分别为2.219±0.055pg/2c和2.214±0.083pg/2c。     

蟹类线粒体DNA的研究与应用

线粒体DNA作为理想的分子遗传标记已被广泛用于蟹类种群遗传学和进化遗传学的研究,并取得了许多有意义的结果。本文阐述了蟹类线粒体DNA分子生物学的研究进展,重点介绍蟹类线粒体DNA序列的研究概况及其多态性在蟹类系统学、种群识别、起源和进化、地理分化等研究中的应用情况。     

双壳纲八种贝类的核DNA定量分析及其与进化地位关系的初步探讨

运用流式细胞仪测定了双壳纲8种贝类的细胞核DNA含量。结果显示,测定贝类的DNA含量分别为(pg):太平洋牡蛎,2.06±0.03;东方海笋,2.11±0.07;近江牡蛎,2.18±0.13;结蚶,2.59±0.10;青蚶,3.08±0.23;砂海螂,3.24±0.28;缢蛏,3.85±0.10;日本镜蛤,5.00±0.20。双因素方差分析表明,同种贝类个体间DNA含量的差异不显著(p>0.05),仅占总差异的0.12%;而种间差异极为显著(p<0.01),比例高达97.45%。本实验所测8种贝类的DNA含量与其进化地位间未呈现出明显的相关性。     

DN A条形码技术及其在海洋贝类种质资源保护中的应用

DNA条形码技术是一种新兴的分子鉴定方法,是指通过选择一段DNA基因序列片段作为条形码进行物种鉴定、系统发育研究和物种多样性保护等。海洋贝类种类丰富、数量众多,传统的形态分类方法有一定的局限性,对一些非常见及疑难物种不能快速、准确地鉴定。作为一种快速、高效的物种诊断技术,DNA条形码技术在海洋贝类种质资源保护与利用方面起到了至关重要的作用。本文就DNA条形码的产生、原理、分析步骤、优势、局限及在海洋贝类种质资源保护中的应用进展进行了综述,并对其未来的发展进行了分析展望。     

Phylogeography of the rocky intertidal periwinkle Echinolittorina paytensis through a biogeographic transition zone in the Southeastern Pacific

Barriers to dispersal are recognized to play an important role in the differentiation of populations and ultimately in speciation. In the southeast Pacific, on the northern coast of Peru, a transition zone between the Peruvian and Panamic marine biogeographic provinces exists. Here, the convergence between two contrasting surface currents could generate a barrier effect for the larval dispersal of meroplanktonic invertebrates, which could in turn generate differentiated populations or genetic lineages on both sides of the transition zone. To address this, we studied to Echinolittorina paytensis, an abundant rocky intertidal periwinkle that spans both biogeographic provinces. A total of 95 individuals from Ecuador (2°19′S) to central Peru (7°31′S), covering the Panamic Province, the Peruvian Province, and the transition zone between, were collected. The mitochondrial markers cytochrome c oxidase I and 16SrRNA were sequenced in order to investigate phylogeography and genetic structuring. In general, no genetic structuring was found across the transition zone, suggesting this biogeographic boundary would not be acting as a barrier in this species. Factors such as a high larval dispersal capability and the occurrence of El Niño–Southern Oscillation events such as El Niño are discussed.     

Genetic analysis of selected Sargassum fusiforme (Harvey) Setchell (Sargassaceae,Phaeophyta) strains with RAPD and ISSR markers

Since the 1980s,Sargassum fusiforme has been cultivated in Zhejiang,South China,and nowadays it becomes one of the important commercial seaweeds in China.With traditions of eating habits in the East Asian countries,this brown alga is used as food,because it contained functional oligo/polysaccharides and chemical components,and was regarded playing roles in antioxidant activities and regulating immunology.Through over 15 years’selection,breeding and cultivation,we obtained three strains with good traits and testified their characters during the production,which included the cultivars with high yield and other two good characters,either all the selected strains were applied in the Sargassum production.To avoid confusion during the selection and nursery,it was preferred to establish one fingerprint for distinguishing the Sargassum cultivars from different strains.Random amplified polymorphic DNA(RAPD)and inter-simple sequence repeat(ISSR)methods were adopted to analyze the genetic diversities of the selected S.fusiforme strains.With that,one fingerprint with RAPD markers was constructed,and one sequence characterized amplifi ed region(SCAR)marker to S.fusiforme was obtained.It is indicated that the applied fingerprint could be valid in S.fusiforme genetic and germplasm justification,and will be positive to molecular marker assistance in its selection and cultivation.     

许氏平鲉4 个野生群体遗传多样性微卫星分析

利用18对微卫星引物分析了荣成湾群体1、荣成湾群体2、即墨群体和龙口群体共4个许氏平鲉(Sebastes schlegeli)野生群体的遗传多样性。结果表明:4个群体的平均有效等位基因数分别为3.1、2.7、2.8和2.7,平均期望杂合度分别为0.590、0.540、0.552和0.491,平均多态信息含量分别为0.577、0.495、0.538和0.528。4个许氏平鲉群体均表现出较高的遗传多样性水平,但也都存在杂合子缺失现象。Χ2检验显示,4个群体中大部分微卫星位点偏离Hardy-Weinberg平衡(P<0.05)。群体间Fst值为0.04,说明遗传变异主要存在于群体内,群体间的遗传分化程度较低。该研究结果将为许氏平鲉种质资源保护和合理利用以及人工增殖提供理论参考。     

广东沿海杂色鲍养殖群体遗传多样性的微卫星分析

实验选取的7个杂色鲍(Haliotis diversicolor)养殖群体分别来自广东汕头、汕尾、惠东、湛江和徐闻,深圳野生驯养,利用7对微卫星引物对上述群体进行了遗传多样性和群体遗传结构的分析。结果显示:7个位点产生的等位基因数从7～13个不等,共检测到68个等位基因,平均每个位点的等位基因数为9.71个。有效等位基因数的范围为3.8～8.1,平均观测杂合度范围为0.422～0.906,平均期望杂合度范围为0.748～0.917。7个群体的多态信息含量变化范围为0.657～0.883,PIC值均在0.500以上,表现为高度多态性。与两个野生驯养的养殖群体相比,其他5个养殖群体均存在不同程度的遗传多样性降低。AMOVA分析显示,4.79%的遗传变异来自于群体之间,95.21%的变异来自于群体内的个体之间。研究表明,广东沿岸的杂色鲍具有较高的遗传多样性水平,7个群体之间呈现一定的遗传分化。     

纤毛虫单细胞基因组DNA微量提取方法的比较及优化

在纤毛虫的分子生物学研究中,单细胞基因组DNA的微量提取是后续基因分析的关键。本研究以海洋纤毛虫红色伪角毛虫为材料,采用直接取完整虫体、裂解法以及3种改良后的试剂盒微量提取DNA方法,分别提取了新鲜样品与4种不同方法保存样品的基因组DNA,比较了9种组合在PCR产物质量、提取所需时间、成本及便捷性方面的优劣,并进一步比较、讨论和探索了4种样品保存方法和5种DNA微量提取方法在纤毛虫单细胞基因组PCR扩增中的可行性。本工作旨在为后续的纤毛虫分子生物学研究提供可资借鉴的技术路线和方法。     

Seasonal Liza aurata tissue-specific DNA integrity in a multi-contaminated coastal lagoon (Ria de Aveiro, Portugal)

In this study, the DNA integrity of golden grey mullet (Liza aurata) collected in differently contaminated sites of a coastal lagoon, Ria de Aveiro (Portugal), was assessed, over the period of 1 year, using the DNA alkaline unwinding assay, in four different tissues (gill, kidney, liver and blood) and compared to a reference site. The four tissues displayed different DNA integrity basal levels, clearly affected by seasonal factors. Gill and kidney were, respectively, the most and least sensitive tissues. All sites demonstrated the capacity to interfere with DNA integrity. The sites displaying the highest and lowest DNA damage capability were, respectively, Barra (subject to naval traffic) and Vagos (contaminated with polycyclic aromatic hydrocarbons). In terms of seasonal variability, autumn seems to be the more critical season (more DNA damage) unlike summer when no DNA damage was found in any tissue. Data recommend the continued monitoring of this aquatic system.