基于贝壳生长轮方法的平端深海偏顶蛤的生长和年龄结构研究

平端深海偏顶蛤(Gigantidasplatifrons)是广泛分布于西太平洋深海热液和冷泉生态系统中的优势种和共有种,也是深海化能生态系统中重要的生境营造种。以南海F冷泉平端深海偏顶蛤为研究对象,基于贝壳日生长轮方法,分析了平端深海偏顶蛤的年龄与生长速率,建立了其年龄与贝壳壳长关系的生长方程。同时,结合获得的生长方程,分析了南海F冷泉采样点(119.285 6°E,22.115 4°N)平端深海偏顶蛤种群的年龄结构。研究结果表明,采样获得的平端深海偏顶蛤种群的最大壳长为11.4 cm,最大寿命为13.5龄。研究区域的贝壳长度分布集中在4～7 cm,占比60%;年龄分布集中在2～4龄,占比49.7%。相关研究结果为进一步开展平端深海偏顶蛤的生长研究提供了基础数据,有助于深入了解冷泉区域的种群动态变化规律。     

平端深海偏顶蛤(Gigantidas platifrons)实验室蓄养过程鳃部含菌上皮细胞超微结构动态变化研究

平端深海偏顶蛤(Gigantidasplatifrons)是南海台西南冷泉区的典型优势物种,鳃丝上皮细胞内共生大量甲烷氧化菌,通过甲烷有氧氧化合成有机物为共生体系提供物质能量,是平端深海偏顶蛤赖以生存的重要能量来源器官,溶酶体在共生体系的营养互作和稳态维持中可能发挥重要作用,本研究使用电镜技术观测了常压培养过程中(0d,30d,90d)平端深海偏顶蛤共生体系中共生菌和溶酶体的动态变化,通过鳃上皮含菌细胞超显微结构的变化研究常压培养对深海共生体系的影响,并探讨溶酶体在宿主—共生菌营养传递和共生菌群稳态维持中的关键作用。研究发现,在原位状态样品中(0d)共生菌和溶酶体呈极化分布,细胞结构完整清晰;蓄养30d后,含菌细胞出现明显破碎,共生菌数量大幅降低,溶酶体数量、范围和消化程度大幅增加,对细胞顶端的甲烷氧化菌进行分解;而90d后,共生菌在溶酶体的作用下消失殆尽,鳃部有明显细胞脱落后留下的坑洞,细胞呈现空泡状,无法明确区分各种细胞组分。上述结果展示了长期常压蓄养过程中鳃上皮含菌细胞中溶酶体与甲烷氧化菌的动态变化,推测当共生菌丢失后溶酶体也同步降低活跃度,平端深海偏顶蛤在共生互作中通过溶酶体主导消化和调控共生菌。     

南海冷泉区贻贝(Bathymodiolus platifrons)附生菌的分离培养与多样性分析

以采集后适应培养0~24 h以及添加甲烷和硫化钠培养24~240 h的南海冷泉区深海贻贝(Bathymodiolus platifrons)为材料,取其鳃部,分析附生菌的多样性与适应性变化状况。共分离鉴定出贻贝附生菌270株,对菌株的16S r DNA进行聚类分析,结果表明所分离菌株主要分布在4个门,21个属,其中变形菌的数量最多且多样性高。分析发现原位新采集的贻贝鳃部附生菌的多样性较高,6 h后附生菌的多样性明显降低。分别添加甲烷和硫化钠对深海贻贝进行培养,甲烷组与碳代谢有关的假单胞菌的数量逐渐增多,且在甲烷组240 h中发现2株食烷菌属菌株。硫化钠组的芽孢杆菌属所占比例升高。此外还发现4株潜在的新种。本研究实现了深海冷泉区贻贝的实验室培养,并对贻贝鳃部附生菌群结构进行了分析,丰富了海洋极端环境微生物资源库,并为深入解析贻贝与其附生菌之间的相互作用关系奠定了基础。     

东太平洋海隆热液区嗜热深海偏顶蛤 （Bathymodiolus thermophilus）的形态和分布特征

在2009-2010年的中国大洋第21航次环球科学考察中,"大洋一号"考察船对东太平洋海隆1.2°N周边新发现的"宝石山"热液区进行了综合调查,并利用我国自主研发的电视抓斗采集到了贻贝类样品,经形态学研究鉴定为嗜热深海偏顶蛤Bathymodiolus thermophilus,Kenk&Wilson,1985,这是此物种在该区域分布的新纪录。文章对样品进行了详细描述,并与其它海区发现的B.thermophilus进行了形态学上的比较和讨论。     

中国常见贻贝基于28S rDNA序列的系统发育分析

贻贝隶属于软体动物门(Mollusca)、双壳纲(Bivalvia)、翼形亚纲(Pteriomorphia)、贻贝目(Mytilida)、贻贝超科(Mytiloidea),大约有400种贻贝分布在世界各地,可适应淡水、潮间带至深海多种生境。本实验以贻贝科6亚科12属28种中国沿海常见贻贝的28SrDNA为目的片段,构建系统发育进化树,运用最大似然法和贝叶斯推演法分析了贻贝科的系统发生,并追踪贻贝科物种系统演化历史。结果显示:贻贝亚科Mytilinae、偏顶蛤亚科Modiolinae、石蛏亚科Lithophaginae均非单系群。在属阶元,深海偏顶蛤属Bathymodiolus、贻贝属Mytilus和股贻贝属Perna为单系群。本研究发现应接受将原隔贻贝属Septifer分为Septifer属和Mytilisepta属的分类提议;应接受将原石蛏属Lithophaga中的膜石蛏亚属Leiosolenus提升至属的地位的分类提议。此外,短齿蛤属Brachidontes的单系性不被支持,刻缘短齿蛤Brachidontessetiger并未与短齿蛤属其他物种在系统发育树上聚拢,亲缘关系较远,为不同属物种,建议恢复刻缘短齿蛤原属名Volsella (Dunker, 1857)。     

深海囊螂蛤铁蛋白的环境选择压力分析

深海环境常有高压、极端温度、富含重金属、黑暗、寡营养等特点。生活在该种特殊环境中的生物可能演绎出不同机制适应此生境。囊螂科贝类是深海冷泉/热液区的优势物种,拟通过对来自冷泉/热液区囊螂科贝类铁蛋白进行环境选择压力分析,以期解析其适应深海特殊生境（高铁环境、高静水压）的机制。首先,通过本地比对的方式获得了囊螂蛤和其近缘种铁蛋白的核苷酸和氨基酸序列,其均有ferritin保守功能域和相应的保守位点。其中,囊螂蛤铁蛋白Cmfer-2、Cmagfer-3、Pofer-3属于M型铁蛋白,其他归属为H型铁蛋白。 Cmagfer-4和Pofer-4可初步判断为分泌型铁蛋白,而其他囊螂蛤铁蛋白与胞质型铁蛋白聚为一支。为了探索环境因素对深海贝类铁蛋白的影响,运用PAML软件对深浅海铁蛋白进行了选择压力分析。 结果显示总共鉴定到8个正选择位点,其后验概率大于95%。其中,在含有嗜压氨基酸的对应组中,约50%浅海位点置换为嗜压的氨基酸,进一步证明了17A、 55R、171S氨基酸置换是铁蛋白适应深海压力变化的一种策略。更进一步研究发现69 K→I/T/M和171 E/K→S由带电荷的极性氨基酸置换为不带电荷的氨基酸,并且大部分受到选择的位点位于蛋白的N端和C端,深海囊螂蛤铁蛋白可能通过特定氨基酸极性的转变和分布区域的改变来适应深海特殊环境。本文通过深浅海贝类铁蛋白的分析来为深海生物对极端环境的适应研究提供一些信息和见解。     

深海化能生态系统双壳纲共生体系互作机制研究进展

双壳纲贝类在深海特殊生境——热液、冷泉及有机沉落生态系统中分布广泛,并且在其体内常含有化能共生细菌为贝类提供营养物质。双壳纲贝类与化能共生菌形成的共生体系对于其适应深海还原性生境至关重要。近40 a来随着海洋科考力度加大,深海化能生境的神秘面纱被逐渐揭开,越来越多的深海物种被发现,双壳纲贝类无疑是这些化能生态系统中的优势物种。在此,我们对常见的深海化能共生双壳纲贝类与其内共生菌的互作研究进行总结,主要双壳纲门类包含贻贝科（Mytilidae）、囊螂科（Vesicomyidae）、蛏螂科（Solemyidae）、索足蛤科（Thyasiridae）和满月蛤科（Lucinidae）,梳理归纳的内容包括深海化能生态系统的发现、“双壳纲贝类—内共生菌”共生体系的组成、共生体系的营养互作、共生体系的建立与维持以及对未来研究的展望。对这些研究内容的总结有利于进一步加深我们对深海特殊生命系统中共生互作机制的认识。     

兰蛤营养成分的研究

对兰蛤的主要营养成分 ,以及氨基酸、脂肪酸、无机元素等的组成进行了研究 ,并同贻贝、蛤蜊、虾夷扇贝、牡蛎进行了比较分析。研究结果表明 ,兰蛤的蛋白质含量仅低于虾夷扇贝 ,其粗脂肪的含量居中 ,碳水化合物的含量最低 ,无机元素的含量最高。兰蛤的氨基酸组成非常全面 ,必需氨基酸总量高于牡蛎的 ,丙氨酸含量居于 5种贝类之首。兰蛤的 EPA和 DHA含量之和明显高于其它 4种贝类。兰蛤中还含有丰富的无机元素 ,Fe、Mn的含量均高于其它 4种贝类 ,其它无机元素的含量也较高     

热液和冷泉10种无脊椎动物基因组大小测定及比较

基因组大小(或称C值)作为生物单倍体细胞中全套染色体的DNA总量,在一定程度上是恒定的,因而C值可以作为生物物种的一个特定参数。深海热液和冷泉为更好地理解C值与不同环境之间的关系提供了一个特征性的模型。本文采用流式细胞术,测定了来自热液和冷泉环境中的10种深海无脊椎动物的C值,其分布范围从0.87 pg到12.28 pg,其中,相比于软体动物和多毛类,甲壳生物基因组大小及变异均较大。对比热液和冷泉两个群落中共有种(深海偏顶蛤Bathymodiolus platifrons、柯氏潜铠虾Shinkaia crosnieri以及长角阿尔文虾Alvinocaris longirostris)的基因组大小,发现C值差异并不显著。同时,综合已有的数据,对深海化能极端环境与其他环境条件下的物种C值进行对比分析,结果显示深海化能极端环境下生物的基因组大小并没有发现明确的变化趋势。     

偏顶蛤（Modiolus modiolus）转录组组装及其与深海偏顶蛤（Bathymodiolus platifrons）比较转录组研究

双壳类适应性进化遗传基础尚未有大量研究。而生活在沿海潮间带和深海环境中的双壳贝类适应性进化的分子机制更鲜有报道。我们首次测序组装了潮间带生活的偏顶蛤Modiolus modiolus的转录组序列。同时与生活在深海热液冷泉喷口的偏顶蛤Bathymodiolus platifrons进行了比较转录组分析，阐明了这两个物种在不同环境中的适应性进化机制。M. modiolus和B. platifrons的转录组组装共产生182，476和156，261个转录本，N50值分别为1，769和1，545。同时注释到27，868和23，588个基因。GO富集分析表明这些基因有相似的富集模式。两物种进化分析鉴定到10，245个直系同源基因，其中26个基因受到强烈正选择（Ka/Ks＞1），12个基因受到中度正选择（0.5M. modiolus的转录组组装拼接序列，同时通过比较转录组学分析阐明了其与近缘物种B. platifrons的适应性进化机制。这为两个物种后续研究提供了序列资源和研究基础。     

印度洋深海热液区贻贝足丝结构和性能特征研究

贻贝营半固着生活,依靠足丝将自己牢固地黏附在底物上.足丝的结构和性能对贻贝的正常生命活动至关重要.为研究热液区极端环境下深海贻贝足丝的结构和性能,采集了印度洋龙旂深海热液区的贻贝(Bathymodiolus marisindicus)足丝,对其力学特性、结构特征和组分进行了分析.力学性能测试发现与近海贻贝(Mytilu...     

Life-history traits of the symbiotic scale-worm Branchipolynoe seepensis and its relationships with host mussels of the genus Bathymodiolus from hydrothermal vents

Associations between scale‐worms and giant mussels are common constituents of hydrothermal vent and cold seep ecosystems, but very little is known about their nature and ecology. Here, we analyze the ecological characteristics of the associations between Branchipolynoe seepensis, an obligate symbiotic polychaete, and their host mytilid mussels Bathymodiolus puteoserpentis and B. azoricus inhabiting hydrothermal vent fields on the Mid‐Atlantic Ridge. Infested mussels generally harbored a single symbiont (<4% had two to six worms). Infestation rate varied from 7.2% to 76.5%, increasing with mussel size, and was significantly lower for B. puteoserpentis. Symbiont density at Lucky Strike ranged between 1071 and 1191 individuals m⁻². Female symbiont size was always positively correlated with host size, while only males and juveniles from small mussels showed the same trend. This suggested a relatively long‐lasting host/symbiont association for females and short‐lasting association with successive reproductive migrations for adult males. The sex ratio of symbionts was always biased in favor of females. Males were smaller and more slender than females and had one mode in their size distributions, whereas females typically had three or more modes, suggestive of a longer life span in females. Between 59.1% and 72.2% of mussels had damaged soft tissues with substantially higher incidence of trauma in infested ones, suggesting that symbionts may cause trauma. The symbionts also induce tunnel‐like structures among the ctenidia, indicating fidelity to a particular location inside the host. Based on our data, together with the fact that infested mussels became relatively wider than non‐infested ones, this association is considered parasitic (likely kleptoparasitic). Our data, together with those from previous studies, allowed us to define the main life‐history traits of B. seepensis: (i) the relationship with their host is parasitic, (ii) the association begins at the smallest mytilid size classes, (iii) there is sexual dimorphism in body size, (iv) sex ratio deviates from 1:1 in favor of females, (v) fertilization occurs through temporal pairing and pseudocopulation, (vi) sperm are stored by females, (vii) eggs are large (likely lecithotrophic or with direct development), (viii) females have a longer life span than males, (ix) adult males may be semalparous, undertaking reproductive migrations followed by a short period of pairing and then death, and (x) females have a semi‐continuous iteroparous reproductive cycle.     

Adaptation of the antioxidant defence system in hydrothermal-vent mussels (Bathymodiolus azoricus) transplanted between two Mid-Atlantic Ridge sites

The vent mussel Bathymodiolus azoricus is the dominant member of the Northern Mid‐Atlantic Ridge (MAR) hydrothermal megafauna, and lives in an environment characterized by temporal and spatial variations in the levels of heavy metals, methane and hydrogen sulphide, substances which are known to increase reactive oxygen species levels in the tissues of exposed organisms. To evaluate the effects of two contrasting hydrothermal environments on the antioxidant defence system of this vent mussel species, a 2‐week transplant experiment was carried out involving mussels collected from the relatively deep (2300 m), and chemical rich, Rainbow vent field. These were transplanted to the shallower (1700 m), and relatively less toxic, Lucky Strike vent field. To achieve this objective, levels of superoxide dismutase, catalase (CAT), total glutathione peroxidase (GPx), selenium‐dependent glutathione peroxidase and lipid peroxidation (LPO) were measured in the gills and mantle tissues of resident and transplant mussels before and after the transplant experiment. With the exception of CAT, the gills of the transplanted mussels had significantly higher antioxidant enzyme activity compared with the basal levels in the donor (Rainbow) and recipient (Lucky Strike) populations; whereas the antioxidant enzyme levels in the mantle tissues of the transplants reflected the baseline levels of activity in the native Lucky Strike mussels after 2 weeks. In contrast, LPO levels were significantly higher in both tissue types in the transplants than in either the source or the recipient populations, which suggested a response to hydrostatic pressure change (note, the transplant animals were brought to the surface for transportation between the two vent fields). The fact that the Rainbow mussels survived the transplant experience indicates that B. azoricus has a very robust constitution, which enables it to cope behaviourally, physiologically and genetically with the extreme conditions found in its naturally contaminated deep‐sea environment.     

软体动物腹足纲分类学研究进展——从近海到深海

随着科学技术的不断进步和研究的不断深入,我国关于海洋的研究已由浅海逐步向深海发展,不断取得新的发现和认知。自2014年4月起,中国科学院海洋研究所依托先进的"科学"号海洋科学综合考察船,搭载"发现"号水下机器人（remotelyoperated vehicle,ROV）,先后对冲绳海槽热液区、南海冷泉区、雅浦海山区以及马努斯热液区等深海区域进行了综合性科学考查,采集到了大量深海腹足纲（Gastropoda）标本,其中有一些为未经发现和描述的新种,极大地推动了我国关于深海软体动物物种多样性研究,实现了深海环境和软体动物资源的新认知。本文对近年来从深海采集到的腹足类标本进行了整理分类,对其中的11属13种进行描述,并对深海特殊环境中腹足纲物种多样性进行了总结,对中国软体动物腹足纲从近海至深海的分类现状进行研究论述。     

Ecophysiological differences between vesicomyid species and metabolic capabilities of their symbionts influence distribution patterns of the deep‐sea clams

This study provides an analysis of vesicomyid bivalve–symbiont community distribution across cold seep and hydrothermal vent areas in the Guaymas Basin (Gulf of California, Mexico). Using a combination of morphological and molecular approaches including fluorescent in situ hybridization (FISH), and electronic microscopy observations, vesicomyid clam species and their associated symbionts were characterized and results were analyzed in light of geochemical conditions and other on‐site observations. A greater diversity of vesicomyids was found at cold seep areas, where three different species were present (Phreagena soyoae [syn. kilmeri], Archivesica gigas, and Calyptogena pacifica). In contrast, A. gigas was the only species sampled across the hydrothermal vent area. The same haplotype of A. gigas was found in both hydrothermal vent and cold seep areas, highlighting possible contemporary exchanges among neighboring vents and seeps. In either ecosystem, molecular characterization of the symbionts confirmed the specificity between symbionts and hosts and supported the hypothesis of a predominantly vertical transmission. In addition, patterns of clams could reflect potential niche preferences for each species. The occurrence of numerous traces of vesicomyid movements on sediments in the sites colonized by A. gigas seemed to indicate that this species might have a better ability to move. Furthermore, variation in gill sulfur content could reveal a higher plasticity and sulfur storage capacity in A. gigas. Thus, the distribution of vesicomyid species across the chemosynthetic areas of the Guaymas Basin could be explained by differences in biological traits of the vesicomyid species that would allow A. gigas to more easily exploit transient and punctual sources of available sulfide than P. soyoae.     

Epsilon-变形菌趋化信号通路中偶联蛋白CheV与趋化受体蛋白的CZB结构域的功能研究

细菌通过趋化系统感知和响应外界环境变化并进行趋利避害, 因此很多细菌进化出了非常复杂多样的趋化系统以适应不同的生态位。Epsilon-变形菌广泛存在于自然界中, 能适应不同的生态环境, 特别是深海Epsilon-变形菌能适应深海热液口和冷泉等极端环境, 其趋化系统可能有其独特之处。通过BlastP和MIST数据库分析, 我们发现大部分深海Epsilon-变形菌拥有F3类型趋化系统且具有单拷贝的双功能域融合蛋白CheV。此外, 一种特殊结构域CZB (C-terminal Zinc-Binding) 存在于深海Epsilon-变形菌的趋化受体蛋白中, 而且在部分深海Epsilon-变形菌的趋化受体中还发现并鉴定了一种新的结构域CZB-like结构域。文章以Epsilon-变形菌的模式菌株空肠弯曲杆菌(Campylobacter jejuni 81-176)为研究对象, 用细菌双杂交试验证实了CheV能与所有含MA结构域的趋化受体相互作用。用电感耦合等离子体质谱(ICP-Mass)技术证实了CZB-like结构域不能结合Zn离子, 但细菌双杂交试验证实它能介导趋化受体Tlp9与CheV的互作。     

Effect of cadmium, copper and mercury on antioxidant enzyme activities and lipid peroxidation in the gills of the hydrothermal vent mussel Bathymodiolus azoricus

Metals are known to influence lipid peroxidation and oxidative status of marine organisms. Hydrothermal vent mussels Bathymodiolus azoricus live in deep-sea environments with anomalous conditions, including high metal concentrations. Although B. azoricus are aerobic organisms they possess abundant methano and thioautotrophic symbiotic bacteria in the gills. The enzymatic defences (superoxide dismutase (SOD), catalase (CAT), total glutathione peroxidase (Total GPx) and selenium-dependent glutathione peroxidase (Se–GPx)) and lipid peroxidation were determined in the gills of B. azoricus exposed to Cd (0.9 μM), Cu (0.4 μM) and Hg (0.1 μM) with different times of exposure. The experiments were performed in pressurized containers at 9 ± 1 °C and 85 bars.Results show that vent mussels possess antioxidant enzymatic protection in the gills. Cd and Cu had an inhibitory effect in the enzymatic defence system, contrarily to Hg. These enzymatic systems are not completely understood in the B. azoricus, since reactive oxygen species might be produced through other processes than natural redox cycling, due to hydrogen sulphide and oxygen content present. Also the symbiotic bacteria may play an important contribution in the antioxidant protection of the gills.