中国沿海无柄蔓足类研究进展

无柄蔓足类属节肢动物门甲壳纲,是海洋生态系统和污损生物群落中极为重要的组成部分,在中国海域分布着6科25属110种,主要种类为纹藤壶(Balanus amphitrite amphitrite)、网纹藤壶(B.reticulatus)、高峰星藤壶(Chirona amaryllis)、泥藤壶(Balanus uliginosus)、白脊藤壶(B.albicostatus)、三角藤壶(B.trigonus)、红巨藤壶(Megabalanus rosa)、钟巨藤壶(M.tintinnabulum tintinnabulum)、白条地藤壶(Euraphia withersi)、鳞笠藤壶(Tetraclita squamosa squamosa),其中纹藤壶在黄、渤海为优势种,网纹藤壶则在热带和亚热带海区占优势;泥藤壶多出现在沿海河口的咸淡水交汇处;三角藤壶、红巨藤壶和钟巨藤壶等种类分布于盐度较高的海域。环境因子可对无柄蔓足类的生长发育、繁殖附着、分布状况及形态特征等产生显著影响。幼虫发育阶段要经历6期无节幼虫和1期金星幼虫,青岛大扁藻(Platymonas helgolandica)、牟式角毛藻(Chaetoceros muelleri)和亚心形扁藻(Platymonas subcordiformis)均是幼虫培养较为理想的饵料;金星幼虫可在4—8℃下保存1周左右。藤壶胶粘物由蛋白亚基聚合而成,其初生胶和次生胶组成基本相似。无柄蔓足类不仅是开展防除测试和生态科学研究的理想材料,而且还应进一步分析其在海洋生态系统中的地位和作用,并从分子水平探讨幼虫附着机理、胶粘物作用机制、种类相互关系与系统发生史。     

饵料、盐度和温度对刺巨藤壶幼虫生长发育的影响

刺巨藤壶(Megabalanus volcano)是沿海污损生物的优势种,也是沿岸居民常食用的海鲜之一。为探究刺巨藤壶幼虫室内培养的适宜条件,采用生态学单因子梯度实验方法,研究饵料种类及密度、盐度(18、22、26、30、34、38和42)和温度(14℃、18℃、22℃、26℃、30℃和34℃)等生态因子对刺巨藤壶幼虫生长发育的影响,以Ⅵ期无节幼虫存活率、Ⅵ期无节幼虫发育时间、金星幼虫存活率、金星幼虫发育时间和金星幼虫体长等为观察指标。中肋骨条藻(Skeletonema costatum)和牟氏角毛藻(Chaetoceros muelleri)均设置5×10⁴个/mL、10×10⁴个/mL、20×10⁴个/mL、40×10⁴个/mL和80×10⁴个/mL 5个密度梯度,亚心形扁藻(Platymonas subcordiformis)设置2.5×10⁴个/mL、5×10⁴个/mL、10×10⁴个/mL、15×10<...     

环境因子对海月水母生长发育影响的研究进展

海月水母是一种广泛分布的世界性近岸种,也是我国近海海域大型水母优势种之一.近年来海月水母在世界各地频繁暴发,对海洋生态系统和沿岸经济、社会发展带来了很大危害.海月水母有复杂的生活史(营附着生活的螅状体世代和营浮游生活的水母体世代)和多种无性生殖(出芽生殖、横裂生殖、足囊生殖等)及有性生殖方式.环境因子影响海月水母生活史的各个阶段,如浮浪幼虫阶段、螅状体阶段、碟状体阶段等.这些生长阶段尤其是早期生长阶段的状态(如附着、分布、扩散等)会进一步影响海月水母的种群动态.本文以海月水母生活史为主线,系统总结分析了国内外关于环境因子对其生活史各阶段的影响,提出了一些值得进一步深入研究的问题,以期为我国近海海域水母暴发关键因子的研究提供借鉴.     

美丽海绵提取物防污损作用

从美丽海绵分离提取了2-去氧-1-氢-1,2,4,三唑、环(脯氨酸-甘氨酸)、尿嘧啶、环(脯氨酸-丙氨酸)、6-氨基嘌呤、4-(1-苯乙基)苯酚、1-氢-1,2,4,三唑和胆甾-5-烯-3β,7α-二醇等8种化合物,并研究了其对网纹藤壶金星幼虫的影响,其中6-氨基嘌呤(7.1 μg/cm2)和4-(1-苯乙基)苯酚(5.9μg/cm2)对金星幼虫具有明显的毒杀作用.美丽海绵可能通过这些活性化合物的协同作用来防止它种生物附着污损.     

人工红树林幼林藤壶危害及防治研究进展

随着近年来红树林恢复性造林面积的扩大,海洋污损生物藤壶对红树林幼林的危害问题日益突出.文中综述了藤壶附着的生物化学、藤壶在红树林附着的生态学、藤壶对人工红树林幼林的危害和国内所采用的化学药物防治措施等方面的研究进展,以及今后的研究方向.藤壶在红树林的附着和分布模式受海水盐度、浸淹深度、林分郁闭度、水文条件等环境因素和生物因素的影响.而藤壶胶粘蛋白的氨基酸组成、一维结构,胶粘蛋白在水下的交联、组装和胶粘的过程与机制,以及藤壶危害红树幼苗的机制和危害权重尚需要深入探讨.研究红树植物对藤壶附着的响应和长期适应机制将为藤壶的防治提供更多的启示.     

蔓足类的生物学

介绍我国沿海岸藤壶的某些生物学特性。繁殖和附着期与附着量和水温密切相关。附着量还与光、颜色、水深和水流有关。外壳的生长分为二个阶段,生长初期外壳基底的直径迅速增长,生长后期壁板增厚,壳重迅速增加。生活史包括６期无节幼虫和１期腺介幼虫。幼体的发育时间与水温密切相关。蔓足类是滤食性动物,以浮游生物和有机碎屑为主。共栖行为在蔓足类中极为普遍,有利于种群的分布和摄食,又可得到宿主的保护。蔓足类的定向与水流     

K~(+)、Cr~(6+)对网纹藤壶幼虫发育和存活的影响

以华南沿海污损生物群落优势种网纹藤壶(Balanus reticulatus)的幼虫为研究对象,探讨钾、铬2种金属离子对其幼虫存活及发育的影响。实验所用金属离子溶液是将K+和Cr6+母液添加到过滤消毒海水中配制。将活泼健壮的网纹藤壶无节幼虫置于上述溶液中,以亚心形扁藻(Platymonas subcordiformis)为饵料于30℃左右的黑暗环境中培养5d,镜检观察记录幼虫发育状况并进行统计分析。结果表明,K+对网纹藤壶幼虫发育和存活的影响均不明显;而Cr6+对幼虫发育具有一定的毒杀和抑制作用,不仅幼虫存活率低于对照组,而且金星幼虫所占百分率也低于对照组。     

海假交替单胞菌fliC-02330基因缺失影响生物被膜形成及厚壳贻贝幼虫附着变态

【背景】假交替单胞菌属是一种广泛分布于海洋环境的革兰氏阴性细菌,存在于海底沉积物中,能分泌大量的胞外产物形成海洋微生物被膜,从而诱导海洋无脊椎动物的附着。【目的】探究海假交替单胞菌鞭毛蛋白fliC基因对生物被膜形成及厚壳贻贝诱导活性的影响。【方法】通过基因敲除构建海假交替单胞菌fliC-02330基因缺失突变菌,研究突变菌和野生菌菌落形态、生物被膜形成能力、胞外物质以及对厚壳贻贝幼虫附着变态的诱导能力等的差异性。【结果】与野生菌相比,突变菌菌落表型出现褶皱,运动能力下降,形成被膜膜厚增加,以及对幼虫附着变态诱导活性下降。共聚焦扫描发现,fliC-02330基因缺失突变菌胞外多糖含量下降,而蛋白含量上升。【结论】海假交替单胞菌鞭毛蛋白fliC-02330基因缺失促进生物被膜形成,但抑制厚壳贻贝幼虫附着变态。本研究为探究细菌鞭毛蛋白基因与厚壳贻贝幼虫的作用机制,以及后续进一步探索微生物参与海洋无脊椎动物附着变态提供一定的理论依据。     

环境因子对大亚湾人工鱼礁上附着生物分布的影响

通过对深圳大亚湾人工鱼礁区7个月(2008-04～2008-10)挂板实验,同时监测相应的环境因子指标.以附着生物丰度数据进行了除趋势对应分析(DCA),并根据附着生物丰度数据和7个环境因子进行了典范对应分析(CCA).共鉴定出附着生物54种,物种鉴定结果表明实验试板上的常见种有华美盘管虫 (Hydroides elegans)、三角藤壶( Balanus trigonus)、细肋肌蛤(Musculus mirandus)等.DCA种类排序图可明显地看出每种附着生物都有自己的分布中心和分布区域;CCA分析结果表明深度、透明度、溶解氧是影响附着生物群落变化的最主要环境因子,其次是盐度和温度.CCA排序图较好地反映了人工鱼礁上附着生物分布与各环境因子的相互关系.     

温度对海月水母浮浪幼虫的影响

海月水母是黄渤海沿岸海域的爆发水母之一, 具有复杂的生活史, 目前关于其浮浪幼虫生活史阶段的研究相对较少。研究了24 ℃(夏末)和19 ℃(秋初)两种温度对海月水母浮浪幼虫的存活时间、附着和形态的影响。研究结果表明: (1)两种温度条件下海月水母的浮浪幼虫的最大浮游存活时间为21-28 天。(2)温度对海月水母浮浪幼虫的死亡率有显著影响(F = 4.238; P < 0.05), 24 ℃下的死亡率显著高于19 ℃。(3)两种温度条件下海月水母浮浪幼虫的附着率无显著差异(P > 0.05)。(4)温度对海月水母浮浪幼虫的长度有显著影响(F = 7.153; P < 0.05), 对海月水母浮浪幼虫的宽度影响不显著(P > 0.05)。上述研究结果为海月水母早期浮浪幼虫生活史阶段种群动态研究提供了重要参考。     

Nature and perception of barnacle settlement pheromones

It is now almost 50 years since the gregarious settlement of barnacles and its chemical basis was first described. Although originally noted for Elminius modestus, mechanistic studies of gregariousness have focused on two species, Semibalanus balanoides and Balanus amphitrite. By virtue of its ease of study and its economic importance as a fouling organism, the latter species has assumed increasing importance in recent years. This paper will provide an overview of studies on settlement pheromones and their perception. An adult glycoprotein, arthropodin (now known as settlement-inducing protein complex or SIPC), was once thought to be the sole pheromone involved in the induction of cypris larval settlement. At least two other pheromones are now known to be involved, a waterborne cue originating from the adult and the cypris temporary adhesive. The latter is related, immunologically, to SIPC. In keeping with many other examples of chemical communication, the available evidence suggests that barnacle settlement induction involves receptor-ligand interactions and a signal transduction pathway(s) that translates into attachment and metamorphosis. Similar findings have been reported for some, but not all, marine invertebrate larvae examined thus far and the implications for antifoulant development are discussed.     

Immunological studies on the settlement-inducing protein complex (SIPC) of the barnacle Balanus amphitrite and its possible involvement in larva-larva interactions.

Immunological investigation has revealed that a settlement-inducing protein complex (SIPC), which induces cypris settlement of the barnacle Balanus amphitrite, is synthesized during larval development and accumulates in the cypris larva. We previously purified the SIPC from adult B. amphitrite, which was active when bound to a substratum. The SIPC is a glycoprotein of high molecular mass, consisting of three major subunits of 76, 88 and 98 kDa with lentil lectin (LCA)-binding sugar chains. In the present study, we prepared antiserum against each LCA-binding subunit of SIPC, and performed immunoblot analyses. Immunoblotting of adult extracts showed that anti-76-kDa antibody reacted only with the 76-kDa protein, whereas anti-88-kDa and anti-98-kDa antibodies reacted with both the 88-kDa and the 98-kDa proteins. Immunoblotting of larval extracts indicated that reactivity of the 76-kDa protein to anti-76-kDa antiserum increased during larval development and cyprid extracts reacted strongly. Moreover, by using immunostaining we found that the SIPC was contained in ''footprints'' of cyprids, which have been shown to act as a settlement-inducing pheromone, and is secreted onto the antennular attachment discs. The results suggest that the SIPC (or SIPC-like proteins) is involved in both adult-larva and larva-larva interactions during settlement of the barnacle B. amphitrite.     

Species specificity of barnacle settlement-inducing proteins

We previously isolated a larval settlement-inducing protein complex (SIPC) from adult extracts of the barnacle, Balanus amphitrite using a nitrocellulose membrane settlement assay. In the present study, we found that the extracts of other adult barnacles, Megabalanus rosa and Balanus eburneus, also induced the settlement of B. amphitrite cyprids although the inductive activity was slightly lower than that of conspecific extracts. Furthermore, we examined reactivity to anti-SIPC antibody in adult extracts from six species of Japanese barnacles other than B. amphitrite, brine shrimp and eight marine sessile organisms besides barnacles. The results showed that all barnacles examined contained SIPC-like proteins with slightly different molecular weight, while the other animals did not react to the antibody by immunoblot analysis. These findings suggest that species specificity in settlement-inducing proteins of barnacles is not so strict, but these proteins are characteristic to barnacle species.     

Discrimination at settlement in barnacles: Laboratory and field experiments on settlement behaviour in response to settlement‐inducing protein complexes

A major driving force to mechanistic studies of barnacle gregarious settlement is to contribute to an understanding of observed patterns of settlement in nature. In particular, how cyprids perceive adult conspecifics and how they discriminate between conspecific and allospecific barnacles are questions which have taxed researchers for nearly 50 years. The putative, active component of adult barnacles to which the cyprids respond has long been known to be a glycoprotein, referred to here as the settlement‐inducing protein complex (SIPC). The present study examines the discriminatory abilities of laboratory‐reared Balanus amphitrite and wild Semibalanus balanoides cyprids at settlement. Using a recently developed nitrocellulose membrane‐choice settlement assay, laboratory studies revealed that both species settled at a significantly higher rate on regions of membrane on which crude conspecific SIPC had been adsorbed compared to untreated regions. Moreover, when offered a choice between conspecific and allospecific SIPC, a trend to greater settlement on the conspecific regions was observed. The membrane assay was also evaluated in field trials using real‐time video footage of cyprid searching behaviour. Of 211 S. balanoides cyprids recorded during exploratory behaviour, only one settled. Exploratory behaviour was, however, clearly associated with regions of the membrane treated with either conspecific or allospecific (B. Amphitrite) SIPC compared to untreated regions. These results are generally in accord with previous reports on the discriminatory abilities of barnacle cyprids and suggest that the membrane assay may be usefully applied to field studies of settlement behaviour.     

Identification of the aromatic L-amino acid decarboxylase (AADC) gene and its expression in the attachment and metamorphosis of the barnacle, Balanus amphitrite

Serotonin and dopamine are involved in the attachment and metamorphosis of cypris larvae of barnacles. Aromatic L-amino acid decarboxylase (AADC) gene, the product of which catalyzes the synthesis of serotonin and dopamine from L-5-hydroxytryptophan and L-3,4-dihydroxyphenylalanine, respectively, was characterized. A DNA clone containing part of an AADC sequence was obtained from the genomic DNA library of the barnacle, Balanus amphitrite. This clone had four putative exons consisting of 226 amino acids with an identity of 63.2% and a similarity of 92.1% with human AADC. Northern blot analysis showed that AADC mRNA was expressed at all stages of barnacles: naupliar larvae, cypris larvae and adult barnacles. Two inducers of larval attachment and metamorphosis; that is, serotonin and extract of adult barnacles, obviously increased the expression of AADC mRNA at an early cypris larval stage. These results suggest that intracellular biosynthesis of serotonin, or dopamine, or both is at least partly involved in the control of the attachment and metamorphosis of cypris larvae.     

Cloning of a novel murine gene Sfmbt, Scm-related gene containing four mbt domains, structurally belonging to the Polycomb group of genes

We cloned six cDNAs by screening cDNA libraries of cypris larvae from barnacles, Balanus amphitrite, and studied their expression by Northern blot analysis. All of them are expressed in the cypris larvae at the settlement stage, but not in the earlier nauplii larvae nor in later adult barnacles. Therefore, we designated them as barnacle cypris larva-specific genes (bcs); bcs-1, bcs-2, bcs-3, bcs-4, bcs-5 and bcs-6. During the process of larval attachment and metamorphosis, the amounts of bcs-1 and bcs-2 mRNAs decreased, whereas the bcs-3, bcs-4, bcs-5 and bcs-6 mRNAs increased. A homology search showed that all cDNAs encode novel peptides containing characteristic amino acid sequences. This study strongly suggests that these bcs gene products are involved in the cypris larval attachment and metamorphosis of barnacles.     

Gregarious settlement in cypris larvae:the effects of cyprid age and assay duration

Gregarious behaviour of marine larvae is perhaps most clearly associated with finding a suitable habitat in a changeable or restricted environment, or with finding other conspecifics with which to mate. Prior work has shown that in settlement assays using cypris larvae of the barnacle Balanus amphitrite, gregarious interactions significantly affected the interpretation of experiments testing the activity of organic settlement promoters and inhibitors. Other studies have also shown effects of cyprid age and pheromone concentration on settlement behaviour. However, the effects of interactions between gregariousness and these two factors are not known. The aim of this study was to test the hypotheses that i) as cyprids age the effects of gregariousness become less apparent, and ii) as the duration of the experiment increases gregarious effects become more apparent, using cypris larvae of B. amphitrite and Balanus improvisus. Three age classes of cyprids were used at six densities in a fully factorial design. For B. improvisus cyprids significant gregarious effects occurred between 3 or more larvae, and although larval age and experiment duration had significant main effects, there were no interactions between these important factors and gregariousness. For B. amphitrite cyprids significant gregarious effects also occurred with 3 larvae per well, though this effect was strongly dependent upon experiment duration. B. amphitrite cyprid sensitivity to conspecific cues does not change with age, although increasing experiment duration and age interact to increase settlement. Differences between species may be due to different thresholds to conspecific larval cues, or B. improvisus cyprids release much more larval temporary adhesive during exploration.     

Structures of six cDNAs expressed specifically at cypris larvae of barnacles, Balanus amphitrite

We cloned six cDNAs by screening cDNA libraries of cypris larvae from barnacles, Balanus amphitrite, and studied their expression by Northern blot analysis. All of them are expressed in the cypris larvae at the settlement stage, but not in the earlier nauplii larvae nor in later adult barnacles. Therefore, we designated them as barnacle cypris larva-specific genes (bcs); bcs-1, bcs-2, bcs-3, bcs-4, bcs-5 and bcs-6. During the process of larval attachment and metamorphosis, the amounts of bcs-1 and bcs-2 mRNAs decreased, whereas the bcs-3, bcs-4, bcs-5 and bcs-6 mRNAs increased. A homology search showed that all cDNAs encode novel peptides containing characteristic amino acid sequences. This study strongly suggests that these bcs gene products are involved in the cypris larval attachment and metamorphosis of barnacles.     

Effect of inducers and inhibitors on the expression of bcs genes involved in cypris larval attachment and metamorphosis of the barnacles Balanus amphitrite

We examined the expression of six barnacle cypris larva-specific gene (bcs) cDNAs (bcs-1, -2, -3, -4,- 5, and -6), the bcs genes, by using Northern blot analysis under various conditions that induced or inhibited cypris larval attachment and metamorphosis. Inducers of larval attachment and metamorphosis, such as a neurotransmitter, tended to increase the expression of bcs mRNAs. All inhibitors of larval attachment and metamorphosis, such as G protein-coupled receptor agonists/antagonists, inhibitors of tyrosine kinase-linked receptors and inhibitors of their signal transduction, suppressed the expression of bcs-6 mRNA alone, but affected differentially other bcs genes. These results strongly suggest that the bcs-6 product plays a key role in triggering the attachment and metamorphosis of cypris larvae into juvenile barnacles. The roles of four late bcs genes (bcs-3,-4, -5 and -6) are discussed.     

Purification and partial amino acid sequence analysis of the larval settlement-inducing pheromone from adult extracts of the barnacle,Balanus amphitrite (=Amphibalanus amphitrite)

A previously undescribed larval settlement-inducing protein was purified from adult extracts of the barnacle, Balanus amphitrite (=Amphibalanus amphitrite). Results of SDS-PAGE indicated that the relative molecular mass of the protein in reduced and denatured form is 31,600 ± 500 kDa, and that it is distinct from the Settlement Inducing Protein Complex (SIPC) which has previously been determined as a larval settlement-inducing pheromone. The N-terminal 33-residue sequence of the intact protein showed no similarity with previously reported proteins in the EMBL/Genbank/DDBJ databases. The purified protein at a concentration of 10 μg ml^?1 induced approximately four times more larval settlement than the control (filtered natural seawater). In addition, results of the assay using both 24-well polystyrene plates and agarose gels indicated that this protein is probably released into seawater and attracts cypris larvae. These results suggest that the purified protein is a waterborne type pheromone which induces settlement of larvae of B. amphitrite.