大叶藻移植方法的研究

为探索适宜的大叶藻(Zostera marina)移植修复方法,2008年10～11月,利用沉子法、枚钉法、直插法、夹苗法和整理箱法,在山东荣成俚岛近岸海域进行了大叶藻移植试验,监测了移植后1个月内大叶藻的生长、存活与渗透压的变化,比较了天然大叶藻和移植大叶藻之间的差异,并分析了移植海区主要环境因子与大叶藻生长与存活之间的关系。结果显示,5种移植方法大叶藻的平均存活率为沉子法(100%)>枚钉法(86.7%)>直插法(66.7%)>夹苗法(20%)>整理箱法(0%);移植大叶藻的平均绝对生长率为沉子法(0.358 cm/d)>直插法(0.242 cm/d)>对照组(0.211 cm/d)>枚钉法(0.083 cm/d)>夹苗法(0.067 cm/d);与天然大叶藻相比,移植后大叶藻根的渗透压显著升高,而茎和叶的渗透压则显著降低(P<0.01);移植后大叶藻的生长与存活和移植海区水流、光照、底质等主要环境因子显著相关。研究结果为研发适宜的低成本大叶藻受损生物群落生态修复技术提供了参考。     

天鹅湖大叶藻种苗补充情况调查

本研究以山东荣成典型潟湖——天鹅湖内的大叶藻(Zostera marina L.)海草床为研究对象,从2015年3月到6月对大叶藻种子萌发形成的幼苗进行进了月度调查,包括春季大叶藻种苗的密度、株高、叶鞘高、叶数、叶宽以及分枝情况。结果表明,天鹅湖大叶藻种子在3月即开始萌发,但萌发量较少,种苗密度较低;4月种子大量萌发,种苗密度不断升高,并达到峰值,为(481.77?303.42)株/m2;5月份种子萌发结束,种苗密度下降,同时开始克隆生长;6月份种苗不断生长,分枝数增多,种苗开始出现有性生殖枝。至此,春季种苗的萌发过程结束,进入有性生殖过程。     

大叶藻形态及生长发育特征

对威海双岛湾大叶藻的形态及生长发育特征进行了实地观察和精细解剖研究。结果表明, 大叶藻的形态特征主要表现为: 营养枝具4～8 片叶, 互生, 具管状叶鞘, 叶片线型; 根状茎匍匐于底泥; 每节生两簇不定根; 雌雄同株异花, 佛焰苞花序, 两雄一雌单性无花被花交互排列于花序轴两侧, 花粉胶质丝状; 瘦果。其主要生长发育特征表现为: 叶和节间的基部具有分生组织, 叶和节间生长主要体现为基部的生长; 腋芽的发育具有明显的窗口期; 先出叶先于侧枝萌发; 实生苗或克隆分株的端枝经过近一年的生长分化为成生殖枝, 开化结果后即死亡; 同一花序内的雌花先于雄花开放, 水中受精, 果实败育率较高。大叶藻营养、生殖器官形态结构的特化和它们生长发育的时序优化充分体现了该种对海洋水生环境的高度适应性。     

大叶藻微粒体GST基因对温度胁迫响应的分子机制

为了分析鳗草（Zostera marina）中微粒体谷胱甘肽S-转移酶在不同温度下的基因表达规律以及响应机制,本文通过在大肠杆菌中表达ZmGST,纯化重组蛋白以及热稳定性分析,以此为进一步阐述Z. marina的种群退化机制并提供理论依据。总之,鳗草中的微粒体GST的热稳定性和其对温度变化的响应决定了其对温度的耐受范围,并进而影响其恢复力。     

曼氏无针乌贼肌肉cDNA文库的构建和表达序列标签分析

以曼氏无针乌贼（Sepiellamaindroni）肌肉组织为研究材料,利用Clontech公司的cDNA文库构建试剂盒构建了曼氏无针乌贼肌肉cDNA文库．对文库质量的分析结果表明：cDNA文库的库容量约为1．2×10。克隆（cfu／dm^3）,重组率达96％,插入片段平均长度大于l000bp．挑取568个阳性克隆进行5’末端测序,其中475条ESTs长度大于100bp,初步拼接后得到200个单基因簇,包括41个重叠群和159个单拷贝EST,冗余度为57．89％．经检索,53条ESTs（占比为26．5％）在BLASTx上无明显的同源性（E值≥1．00×10^-10）,为新基因．147条ESTs（占比为73．5％）与已报道的基因有较高的同源性;其中与能量相关基因的ESTs丰度最高,占总数的23．0％．细胞信号传导、细胞骨架和蛋白质代谢的次之,比例分别为17．5％、12．5％、9．5％,其中包括热激蛋白70、转铁蛋白等．这些EST数据有助于进一步筛选和克隆曼氏无针乌贼和其他头足动物的肌肉特异性表达基因．     

桑沟湾楮岛近岸海域大叶藻生态学特征的基础研究

2010年8月至2011年9月期间,对桑沟湾楮岛沿海的大叶藻(Zostera marina L.)进行了一周年调查.结果显示,大叶藻周年平均株高变化范围为(16.97±5.99)~(87.60±20.68)cm,大叶藻平均密度为(613±201)株/m2,单株生物量为0.97~5.31g/株,单位面积平均生物量为594.61~3255.03 g/m2,大叶藻高度和湿重的周年生长变化与水温的变化趋于一致,呈正相关性;大叶藻生长环境水温为5.3~25.6℃,平均温度16.3℃;盐度为28.9~31.3,平均盐度30.6;底质类型为砾砂.大叶藻根茎、叶鞘和叶中碳含量的平均值分别为32.68%±1.27%、33.78%±3.06%、37.01%±1.86%,氮含量平均值分别为1.62%±0.63%、2.79%±0.81%、3.10%±0.81%,磷含量的平均值分别为0.28%±0.04%、0.51%±0.10%、0.48%±0.07%.     

Influences of diel and tidal cycles on fish assemblage in eelgrass (Zostera marina) bed of southern Korea during autumn

Short-term tidal and diel variations of autumn fish assemblage in a Zostera marina bed were investigated using 3 h interval samplings for 24 h in both spring and neap tide using a small beam trawl. A total of 1 346 fishes belonging to 19 species were collected at spring tide, whereas 1 115 fishes belonging to 17 species were at neap tide. The common fish species were Nuchequula nuchalis, Acanthogobius flavimanus, Takifugu niphobles, Acentrogobius pflaumii, and Pholis nebulosa with the former three species dominating at spring tide, while the latter two species being abundant at neap tide. Diel variation in abundance was significant with higher abundance at night than day, but there were no significant differences between spring and neap tides, and between ebb and flood tides (three-way ANOVAs). Diel variation in the abundance of fishes may be influenced by tidal range and cycle, and day-night differences of food availability and behaviors of fishes indirectly. Non-metric multidimensional scaling (nMDS) ordination and analysis of similarity (ANOSIM) results revealed significant differences in species compositions both between day and night, and between spring and neap tide. Eelgrass beds are highly productive marine ecosystem, and thus, our results will contribute to conservation of seagrass ecosystem in the study area.     

盐度对大叶藻种子萌发和幼苗发育的影响

大叶藻海草场是重要生境但在我国严重退化,亟需研究有效的修复方法。本文以桑沟湾的大叶藻种子为研究对象,探讨了盐度急变(0~30)对种子萌发、萌发后继续发育和存活及幼苗幼叶的光合与呼吸速率的影响,为以种子繁殖修复大叶藻海草场提供参考。结果表明:盐度降低促进种子萌发,盐度15及以下效果显著,盐度0时萌发率最高;盐度0~15中萌发的种子均可发育成幼苗,发育和衰亡的比例和速率受萌发盐度背景和萌发后培养盐度的双重影响——较高萌发盐度有利于种子萌发后的发育和存活,种子萌发后于盐度5~30中均可建成幼苗并存活、但在盐度0或1中发育至不同程度后全部死亡。盐度5~30范围内,幼苗幼叶均有净产氧,光合和呼吸作用及净产氧率具随盐度升高而增强趋势。于盐度0或5中萌发、盐度5~30中进一步培育的变盐育苗策略,最高实现了24%或12%的种子于2个月内建成幼苗并继续发育,可作为种子繁殖法修复大叶藻海草场的技术参考。     

基于人为调控盐度的促进大叶藻种子萌发及建苗技术研究

Vast declines in Zostera marina seagrass beds demand effective methods of rehabilitation. In this study, we developed a practical method by reducing salinity to induce seed germination followed with recovering salinity to facilitate seedling production of Z. marina. The results showed that Z. marina seeds collected from natural seawater(salinity 30) were induced to germinate at reduced salinities. Percent germination(GR) was higher and mean-time-to-germinate(MTG) was shorter at lower salinities. The highest GR and shortest MTG occurred at salinity 0(deionized freshwater). After germination in freshwater, seeds could develop into seedlings at salinities5–30 and continue the growth. Viability or development of germinated seeds was not significantly different during the 40 d of post-germination incubation at salinities 5–15 after 1–20 d of germination in freshwater. However,during the process of translating germinated seeds from salinity 0 and 5 to salinity 30, reducing the gradients of post-germination acclimation facilitated more seeds forming seedlings in less time. On average, after 60 d of static incubation, including 20 d in freshwater for germination followed with immediate shift to salinity 5 and increasing to salinity 30 at increment of 5 every two days until cultivation at constant salinity 30, 33% of Z. marina seeds produced healthy seedlings. The results indicate that the salinity-manipulation based method of artificial germination and seedling production is practical and effective in supporting rehabilitation of Z. marina bed.     

大叶藻海草床的生态恢复: 根茎棉线绑石移植法及其效果

于2009年和2010年在青岛汇泉湾用根茎棉线绑石法进行大叶藻(Zostera marina L.)移植, 并于2012年4月20日至11月19日对移植大叶藻的生长情况进行观察(包括形态学变化、密度、茎枝高度、地上生物量、底质粒径几个方面)。观察期间水温为7.8～26.1℃。结果显示, 移植底质可定性为粉砂质;移植大叶藻的有性繁殖期为2012年4～8月; 无性繁殖在秋季达到高峰; 密度在6月和9月分别高达411茎枝/m²和481茎枝/m²; 高度与地上生物量的最大值出现在6～7 份。与2009年青岛湾天然大叶藻进行比较后可以发现, 移植大叶藻的高度、地上生物量及其季节变化与天然大叶藻基本保持一致,说明移植大叶藻的生长状况良好, 同时说明根茎棉线绑石法是一种高效且实用的海草床生态恢复方法。     

Habitat characteristics and spatial arrangement affecting the diversity of fish and decapod assemblages of seagrass (Zostera marina) beds around the coast of Jersey (English Channel)

Recent research and management plans for seagrass habitats have called for landscape level approaches. The present study examines the spatial utilisation of subtidal seagrass beds by fish and decapods around the coast of Jersey (49°N 02° W). A hierarchical scale of landscape configuration and the plant characteristics of eight seagrass beds were measured and the contributions of these variables as predictors of the properties of the fish and decapod assemblages were evaluated using multiple linear regression models. The results indicated that total diversity had a negative relationship with transect heterogeneity and total species number had a weak negative association with increasing fragmentation. Both total diversity and total species number showed a positive relationship with depth. In fact, in all models of species number and densities, values were higher in deeper seagrass beds. Total decapod density increased with aggregation of seagrass patches within a landscape. In addition to landscape configuration, smaller-scale structural changes in both canopy height and epiphyte load appeared to influence densities of decapod crustaceans. At night, fewer patterns could be explained by the independent variables in the model.     

青蛤(Cyclina sinensis)cDNA文库构建及免疫相关因子基因的筛选

在鳗弧菌侵染下,利用SMART方法构建了青蛤的cDNA文库,并采用高通量测序方法和BLASTX比对筛选出ESTs及免疫相关因子的基因.结果表明,所构建的cDNA文库的库容量为1.12×106,插入片段均在500bp以上;大于100bp的有效ESTs序列1113条,平均长度725bp,拼接后获得一致性序列420条,其中包括126个叠联群和294个单拷贝EST,BLASTX比对后获得注释序列271条,进一步筛选获得青蛤免疫相关因子基因序列45条,为克隆其免疫防御相关因子的基因提供了重要的基础.     

大叶藻碎屑作为刺参食物来源的实验研究

采用室内模拟实验方法研究了刺参(Apostichopus japonicus Selenka)与大叶藻(Zostera marina L.)碎屑之间的营养关系, 将大叶藻碎屑与泥质沉积物按照一定的配比作为饵料投喂刺参, 测定刺参的特定生长率和排粪率。按照添加大叶藻碎屑比例的不同, 实验分为5 个处理组, 分别为ES0、ES10、ES20、ES40 和ES100, 大叶藻碎屑含量分别为0%, 10%, 20%, 40%和100%。结果表明, 饵料组成显著影响刺参的生长, 大叶藻碎屑与泥质沉积物混合物中的有机质含量为17%～20%, 水温为13～17℃, 刺参的生长效果较好, 最大特定生长率为1.54%/d, 最大排粪率为1.31g/(个· d)。海草床为刺参提供栖息地的同时, 大叶藻脱落腐败后的有机碎屑可以为刺参提供重要的食物来源, 这对于刺参的资源恢复和营养生态学方面的研究有着重要的意义。     

敌百虫对两种赤潮微藻生长的毒性效应研究

敌百虫是使用最为广泛的有机磷农药之一,在水产养殖中主要用于防治甲壳类寄生虫。本文研究了72 h的急性暴露以及25 d的亚慢性暴露下,敌百虫对两种赤潮微藻海洋卡盾藻和锥状斯氏藻生长的影响,以了解敌百虫对海洋微藻的毒性及海洋初级生产力的影响。在急性暴露下,高质量浓度敌百虫能明显抑制两种藻细胞的生长,而低质量浓度则对藻细胞的生长影响不明显,敌百虫对海洋卡盾藻和锥状斯氏藻生长的72 h EC50分别为32.1 mg/L和22.1 mg/L。在亚慢性暴露下,两种藻细胞对敌百虫的敏感性相近,高质量浓度组(25 mg/L)的藻细胞在暴露的第7~9天就全部死亡;1 mg/L的低质量浓度组,藻细胞的生长状况与对照组相近;5 mg/L和10 mg/L质量浓度组在暴露初期对藻细胞生长具有抑制作用,但在暴露后期则明显刺激了藻细胞的生长。本研究说明水环境背景质量浓度下的敌百虫不会抑制浮游植物的生长,但施药一段时间后,可刺激藻细胞的生长,从而增加了引发赤潮的风险。     

Expressed sequence tag analysis and cloning of trehalose-6-phosphate synthase gene from marine alga Laminaria japonica (Phaeophyta)

A high quality cDNA library was constructed from the brown alga Laminaria japonica,with the titer of 1.2×10 5 pfu/ml.The average insert size of the cDNA library is about 1.6 kb.From the cDNA library,591 cDNA clones were randomly selected and sequenced.As a result,574 EST(expressed sequence tag) sequences were generated.All of the 574 ESTs were submitted to the dbEST database section of GenBank with the accession numbers from CX942625 to CX943198.The cDNA library was screened with a α-32 p labeled 453 bp T P S gene probe,which is a partial sequence yielded from Porphyra yezoensis.Four positive cDNA clones were screened and the sequencing data showed that these four cDNA clones covered majority of L.japonica TPS cDNA sequence.After PCR amplification,sequencing and assembling,the entire ORF(open reading frame) sequence of the T P S gene was obtained,which was named LjTPS.LjTPS encodes a protein containing 908 amino acids with a calculated molecular mass of 101 674 Daltons.The LjTPS gene was successfully expressed in E.coli and rice.The LjTPS gene has potential application both in plant breeding to stress tolerance and in deciphering the T P S gene function and mechanism to stress tolerance.     

雨生红球藻铁型超氧化物酶基因的克隆与序列分析

利用已报道的FeSOD保守区域,设计简并引物,通过简并PCR的方法获得了雨生红球藻(Haematococcus pluvialis)FeSOD(HpFeSOD)cDNA的部分序列,然后采用RACE方法分别克隆到5′端和3′端。拼接后得到HpFeSOD cDNA全长,该基因全长为1 138 bp,ORF为684 bp,编码227个氨基酸。把已经得到的HpFeSOD序列推导成氨基酸序列与一些已知物种的FeSOD氨基酸序列相比较,雨生红球藻FeSOD与杜氏藻、衣藻、团藻、石莼、颤藻的同源性为89%、83%、78%、70%和63%。分子系统学分析表明,雨生红球藻FeSOD与杜氏藻FeSOD聚在一起,介于真菌与高等植物之间并且真核微藻FeSOD与高等植物的同源性更高,推测其在进化上与高等植物的亲源关系更近。     

Seasonal dynamics of Zostera noltii hornem. In the bay of arcachon (France)

Seasonal dynamics of Zostera noltii was studied during 1984 in Arcachon Bay, France. In this Bay, Z. noltii colonizes 70 km², i.e. approximately 50% of the total area, while Z. marina occupies only 4 km². Densities and length of vegetative and generative shoots and above-ground and below-ground biomasses were monitored in four meadows which differed according to their location in the Bay, tidal level and sediment composition. Three of these meadows were homogeneous, well-established beds whilst the fourth was under colonization and patchy. Shoot densities and maximal below-ground biomass were lower in the inner silty seagrass bed than in the sandy meadows located in the centre of the Bay. Maximal above-ground biomasses were similar in the two population types. In the well-established beds, vegetative shoot densities, above-ground and below-ground biomasses showed a unimodal pattern with minima in winter (4000 to 9000 shoots·m⁻², 40 to 80 g DW·m⁻², and 40 to 60 g DW·m⁻², respectively) and maxima in summer (11000 to 22000 shoots·m⁻², 110 to 150 g DW·m⁻², and 140 to 200 g DW·m⁻², respectively). Reproductive shoots were observed from the beginning of June until the end of September, except in the colonizing bed where they persisted until December. Furthermore, in the latter, maximal reproductive shoot density was higher (2600 shoots·m⁻²) than in the established beds (650 to 960 shoots·m⁻²). The total production of Z. noltii in Arcachon Bay was estimated to approximately 35.6·10⁶ kg DW·y⁻¹ (19.4·10⁶ kg DW·y⁻¹ for above-ground parts and 16.2·10⁶ kg DW·y⁻¹ for below-ground parts).     

富营养化和海水盐度对日本鳗草生物量和碳氮含量的协同影响

营养盐和盐度对海草的生长和生理都具有重要影响,但二者对海草的协同作用还未见报道。本研究以日本鳗草(Zosterajaponica)为对象,在室内可控条件下进行营养盐和盐度对日本鳗草的协同影响研究。实验结果显示,营养盐和盐度对日本鳗草地上生物量、地上部分C含量、地下部分C含量均有显著的协同影响。日本鳗草组织中的营养盐含量(Nt)与盐度有显著相关性,但并未受到营养盐和盐度的协同影响。营养盐和盐度会协同影响海草的固碳能力,并同时影响其对硝酸盐的吸收。     

Negative effects of blue mussel (Mytilus edulis) presence in eelgrass (Zostera marina) beds in Flensborg fjord,Denmark

The effect of blue mussel (Mytilus edulis) presence in eelgrass (Zostera marina) beds was studied from June 2004 to July 2005 in Flensborg fjord, Denmark. The field experiments were conducted at two stations, one with only Z. marina (Eelgrass station) present and one where M. edulis were present in the Z. marina beds (Mixed station). Zostera marina parameters were measured (growth of leaves, shoot density, leaf length, and nutrient content) in combination with epiphyte cover and sediment parameters (sulphate reduction rates, sediment nutrient fluxes, organic content, C, N and P content) to examine possible positive and negative effects of the mussels on eelgrass performance. The fluxes of ammonium from the sediments were stimulated at all sampling dates at the Mixed station, and possibly stimulated epiphyte growth at this station. Further ¹⁵N signals in epiphytes from the Mixed station suggested that excretion products from the mussels were important nitrogen sources at this station. Sulphate reduction rates were enhanced at the Mixed station and also sediment sulphide concentrations increased under mussel influence, which may have resulted in sulphide toxicity and decreased growth of Z. marina at this station. The study indicates that for Z. marina beds in Flensborg Fjord the effects of M. edulis in seagrass beds are primarily negative, and raises the question whether this leads to negative effects on the stability and expansion of Z. marina beds.     

缢蛏(Sinonovacula constricta)EST-SSR 分布特征及引物开发利用

采用CAP3软件对NCBI上的5296条缢蛏ESTs序列进行了微卫星特征分析。结果表明,经拼接、去冗得到非冗余EST序列3453条,含SSR位点的EST序列267条,共307个SSR位点,检出率为8.89%,平均每6.83kb出现1个SSR位点。设计了40对EST-SSR引物并进行PCR扩增,29对引物能扩增出理想的PCR产物,其中多态性引物14对。利用14对多态性引物分析了乐清湾缢蛏遗传多样性,共检测到等位基因数(Na)61个,每个位点的等位基因数为2—12个。二核苷酸、三核苷酸和四核苷酸重复是最主要的重复类型,分别占15.96%、37.13%和35.50%。乐清湾缢蛏群体观测杂合度(Ho)、期望杂合度(He)和多态信息含量(PIC)分别为0.569、0.490和0.449,表明乐清湾缢蛏遗传多样性较丰富。