刺参肌肉组织cDNA文库的构建

用RNA提取试剂--TRIZOL Reagent提取刺参(Apostichopus japonicus)肌肉组织总RNA,用SMART cDNA Library Construction Kit构建cDNA文库.经测定原始文库滴度达到 3.2×10~6,扩增后文库滴度达到 5.1×10~9,重组率达到96.7%,从扩增文库随机挑取12 个克隆进行PCR 扩增鉴定,结果显示,插入片段大小为0.5～2.5 kb.通过各项指标验证成功构建了刺参肌肉组织cDNA 文库.     

龙须菜四分孢子体cDNA文库的构建

为了研究不同世代和性别基因表达谱的差异 ,本文构建了龙须菜 (Gracilarialemaneiformis)四分孢子体的 c DNA文库。总 RNA提取使用 RNeasy Plant Mini Kit(Qiagen) ,c DNA文库构建使用 SMART c DNA Library Construction Kit(Clontech) ,包装蛋白购自 Promega公司。该c DNA文库含有 1 .2 8× 1 0 6个克隆 ,扩增文库的滴度是 1 .98× 1 0 9pfu/ m L,重组频率是 85.0 % ,插入片段几乎全部集中在 50 0～ 1 50 0 bp之间     

中国对虾6种组织cDNA文库的构建

以中国对虾血液、眼柄、卵巢、雌虾头胸部、雄虾头胸部和三倍体对虾头胸部组织为材料,采用异硫氰酸胍-酸酚法提取总RNA;用磁珠法纯化mRNA;用Uni ZAP cDNA合成试剂盒合成双链cDNA并进行修饰,将cDNA定向连接在UniZAP载体上;经Gigapack Gold Package包装试剂盒包装成为噬菌体颗粒,转染宿主XL1 Blue MRF’菌株细胞,形成初级文库.初级文库经转染进一步扩增,形成稳定的cDNA文库.6个文库的库容在0.2×106~1.3×106之间,重组率都超过90%.从各文库随机取出6~10个清晰的噬菌斑进行PCR扩增,用琼脂糖凝胶电泳检测,其插入片段长度为500~2500bp.由初步功能基因克隆获得阳性结果.多项指标表明,所构建的对虾cDNA文库质量较高,为进一步筛选目的基因、EST测序和制作基因芯片提供了有效的工具.     

副溶血弧菌感染的九孔鲍血细胞均一化全长cDNA文库的构建

采用Trizol试剂提取副溶血弧菌感染的九孔鲍血细胞的总RNA,经Oligotex纯化得到mRNA.根据SMART技术原理合成双链cDNA.双链cDNA采用双链特异核酸酶进行cDNA的均一化,构建成九孔鲍血细胞的均一化全长cDNA文库.原始文库的库容为3.4×106 cfu/cm3,重组率为92.3%,扩增后文库的滴度为2.6×1011 cfu/cm3以上.从文库中随机挑取897个克隆,测序获得高质量ESTs814条,其中有23条Contigs; 762条Singlets,Unigenes共785条,冗余率只有3.56%.以上结果说明所构建的文库质量好,完全可以满足后续基因克隆和表达序列标签测序工作的需要.     

文昌鱼头部cDNA文库的构建

采用cDNA合成技术制备文昌鱼头部cDNA基因文库.以mRNA为模板,用NotI primer-Adaptor为引物,在反转录酶的作用下,合成第一链cDNA,进而合成第二链cDNA.含有EcoRI和NotI粘性末端的双链cDNA在T4DNA连接酶作用下与λgt11 EcoRI/NotI臂相连,包装蛋白包装,得到噬菌体颗粒,即原始的文昌鱼头部cDNA文库.经X-Gal/IPTG板测定, 该文库滴度为7.8×105 pfu/cm3,重组率为70%.cDNA文库可直接应用于目的基因筛选.     

大叶藻形态特征与显微结构

大叶藻(Zostera marina)是陆生植物起源,在海洋中进行沉水生活的高等单子叶植物。本文主要研究大叶藻的形态特征和组织结构与适水性生活的相关性,通过制作、观察大叶藻石蜡切片,介绍大叶藻的根、茎、叶及种子的显微结构特点。结果发现大叶藻的形态特征和显微结构与沉水生活具有高度的适应性。     

移植大叶藻提高池养对虾产量的研究

本实验首次用移植的方法在虾池内栽培大叶藻。结果表明,大叶藻可以净化虾池水质,改善虾池底质条件,丰富了饵料生物,促进了对虾的生长,提高了产量,每亩虾池的产量提高26.6％,纯利润提高190.5％。文章讨论了提高产量的原因。     

刺参(Apostichopus japonicus)高温胁迫消减cDNA文库的构建与分析

采用抑制性消减杂交技术(SSH)研究刺参高温胁迫下差异表达的基因.分别以高温实验组为检测组(tester)、常温对照组为驱动组(driver),进行正向抑制性消减杂交;以常温组为tester、高温组为driver,进行反向消减杂交,成功构建了刺参正反双向差异消减文库.从两个文库随机挑选384个白斑克隆进行斑点杂交进一步...     

Temporal dynamics of subtidal Zostera marina and intertidal Zostera japonica on the southern coast of Korea

The temporal dynamics of two seagrass species, Zostera marina and Z. japonica, were monitored monthly in Dadae Bay, Geoje Island, on the southern coast of Korea. Plant morphological characteristics, shoot density, biomass, leaf production, reproductive effort, and environmental characteristics were monitored from July 2001 to July 2002. Zostera japonica occurred in the intertidal zone and Z. marina occurred in the subtidal zone from 0.5 to 2.5 m below the mean low water level. Shoots and rhizomes were significantly larger in Z. marina than in Z. japonica, whereas the shoot density was greater in Z. japonica than in Z. marina. Despite differences in morphology and shoot density, biomass did not differ significantly between the species. Reproduction occurred from April to June in Z. marina and from May to July in Z. japonica. The proportion of reproductive shoots was approximately three times higher in Z. marina than in Z. japonica. Seasonal variation in the biomass of Z. japonica was caused by changes in both shoot size and density, whereas that of Z. marina was mainly caused by changes in shoot length. Leaf production in Z. marina and Z. japonica showed clear seasonal variation, and leaf production in Z. marina (2.6 ± 0.2 g DW·m⁻²·day⁻¹) was higher than that in Z. japonica (1.7 ± 0.2 g DW·m⁻²·day⁻¹). The mean plastochrone interval was not significantly different between the two species, whereas the leaf lifetime of Z. marina was longer (69 ± 7.8 days) than that of Z. japonica (59 ± 8.3 days). Our results indicated that seasonal leaf growth patterns in Z. japonica are correlated with irradiance and temperature, whereas those in Z. marina respond most to irradiance. Seasonal changes in irradiance appeared to control the temporal variation in above‐ground biomass in both species.     

A genome-wide analysis of the chloroplast NADH dehydrogenase-like genes in Zostera marina

The chloroplast NADH dehydrogenase-like(NDH)complex,homologous to respiratory complex I,participates in photosystem I cyclic electron flow(PSI-CEF)and chlorores...     

大叶藻(Zostera marina L.)胚的结构和种子萌发过程的研究

以采自山东荣成褚岛桑沟湾南岸成熟的大叶藻种子为材料,研究了种子的外部形态特征、胚乳成分及胚的解剖结构,并初步了解了大叶藻种子的萌发过程。结果表明,大叶藻种子是单子叶有胚乳的种子,呈暗褐色,椭圆形或长圆形,种皮革质且有17～20条纵肋,胚乳丰厚坚硬,胚乳细胞被淀粉粒充满;胚弯曲,由胚根、胚轴、胚芽和子叶组成,子叶U形棒状,其弯曲部有一裂缝,胚芽位于裂缝处,胚轴较短,其下是两个对生的胚根,有中央维管束贯穿胚根、胚轴、胚芽和子叶。在子叶出土萌发的植物类型中,大叶藻种子萌发过程是罕见的。萌发初期,子叶优先生长突破种皮而长出地面,此后胚根才开始生长并产生不定根。对大叶藻种子结构和萌发过程的研究,为探讨大叶藻种子萌发和幼苗更新的限制因素提供了一定的理论依据。     

Influence of the seagrass,Zostera marina L., on current flow

A salt-water flume was used to describe the mechanics of current flow around an articial Zostera marina meadow. Shear velocity and roughness height were positively correlated with seagrass surface area, and were positively/negatively correlated with current velocity. Current velocity intrusion into the meadow before diminution and maximum reduction (both at the 2 cm height line) proceed by factors of 1·25 and 2·07 cm into the meadow per cm s^?1 of current velocity, respectively.Froude number was correlated with mean bending angle of the canopy as a whole. Maximum bending had occurred with Froude = 1, but most bending had taken place by Froude = 0·4, a velocity of 40–50 cm s^?1 in this experiment.The meadow edge is the most dynamic zone of a seagrass meadow in regard to current flow. Bending of the shoot canopy is a mechanism for re-direction of current flow and in-canopy reduction of current velocity. Meadow dimensions may be regulated by scouring processes in different hydraulic regimes. Shoot bending and subsequent in-meadow current velocity reduction are mechanisms that affect self-shading and photosynthetic capabilities as well as providing habitat stability.     

Diagenesis of Organic Matter in Eelgrass (Zostera marina L.) Vegetated Sediments

Oceanology - In February and July 2014, multidisciplinary geochemical studies of the sediments were carried out at two stations. One of them was located in Voevoda Bight, which has a bottom depth...     

Photosynthetic and metabolic responses of eelgrass Zostera marina L. to short-term high-temperature exposure

In recent years, extreme heat events have occurred worldwide and the ocean temperature has been rising, causing stress on the photosynthesis and growth of seagrass. Metabolomics enables detection of metabolic changes under environmental stress. In this study, the photosynthetic physiology and metabolic changes of the eelgrass Zostera marina L. in response to 48 h exposure to 32°C were investigated. The results showed that high temperature induced signi?cant inhibition of photosynthetic effciency(Δ F/F′ m)(23.9%lower than the control), enhanced respiration(58.3% higher), and decreased carbohydrate decomposition products and tricarboxylic acid(TCA) cycle intermediate products, indicating that the energy supply of the eelgrass may be insuffcient at high temperature. In addition, high temperature decreased stearic acid and linoleic acid in eelgrass, suggesting the composition of the membrane system of eelgrass may change at high temperature and implying that high temperature may cause the membrane system to be unstable.     

The role of current velocity in structuring eelgrass (Zostera marina L.) meadows

Measurements of velocity profiles, bathymetry, and surface sediment characteristics across eelgrass (Zostera marina L.) meadows yielded information on community development processes and functional attributes of this ecosystem. Height/length ratios of the meadows were positively correlated with tidal current velocity. Low, medium, and high current regimes were separated by surface current velocities of approximately 50 and 90 cm s^?1. Z. marina can tolerate approximately 120–150 cm/sec current velocities in the areas studied. Per cent silt-clay and organic matter content of the surface sediments are negatively associated with shear velocity, suggesting that meadows in high current areas are sources while meadows in low current areas are sinks of autochthonous detritus. Current velocity maintains seagrass meadows at different equilibrium levels (relative climaxes). We theorize these different equilibrium levels provide unequal habitat utilization potentials for the associated faunal community.     

Nitrogen budget of the eelgrass, Zostera marina in a bay system on the south coast of Korea

Above- and below-ground productivities and tissue N content were measured monthly to quantify N incorporation to sustain eelgrass growth in Koje Bay on the south coast of Korea from January to December 2002. N acquisition was also estimated through measurements of N uptake kinetics, tissue biomass, and in situ inorganic N concentrations in water column and sediments. Above- and below-ground productivities were highest in summer and lowest in late fall and winter. Leaf tissue N content was highest in December and lowest in July, while rhizome tissue N content was highest in October and lowest in April. Estimated monthly N incorporation by leaf tissues based on the leaf productivity and N content ranged from 0.4 g N m^?2 month^?1 in November to 2.0 g N m^?2 month^?1 in May. N incorporation by below-ground tissues ranged from 0.1 g N m^?2 month^?1 in February to 0.2 g N m^?2 month^?1 in October. Annual whole plant N incorporation was 14.5 g N m^?2 y^?1, and N incorporation by leaf tissues accounted for about 87 % of total N incorporation. Maximum uptake rate (V _max ) and half saturation constant (K _m ) of leaf NH₄ ⁺ uptake were significantly lower than those of root NH₄ ⁺ uptake. Above- and below-ground biomass ranged from 20.8 g DW m^?2 and 8.6 g DW m^?2 in winter to 350.0 g DW m^?2 and 81.3 g DW m^?2 in spring, respectively. NH₄ ⁺ concentrations varied from 0.2 to 4.3 mM in water column and from 93.0 to 551.7 mM in sediment pore water. Based on these measurements, annual N acquisition by root tissues contributed slightly higher than that by leaf tissues to total plant N acquisition. During winter, monthly leaf N acquisition was lower than monthly leaf N incorporation. This implies that Z. marina has internal nitrogen retention system to offset the shortage and excess of nitrogen.     

Spatial patterns of epifaunal communities in San Francisco Bay eelgrass (Zostera marina) beds

Epifaunal invertebrate species, such as amphipods and isopods, have been shown to play key but varying roles in the functioning of seagrass habitats. In this study, we characterized patterns in the poorly known epifaunal communities in eelgrass (Zostera marina) beds in San Francisco Bay as a first step in understanding the individual and collective importance of these species, while testing predictions on spatial patterns derived from previous studies in other regions. Surveys conducted at five beds across multiple time periods (April, June, August and October 2007) showed that San Francisco Bay eelgrass beds varied strongly in epifaunal community composition, total, and relative abundance, and that abundance differed markedly among time periods. In contrast to findings by others, morphologically complex flowering shoots frequently harbored greater numbers of epifauna (>2× and up to 10× more individuals) than vegetative shoots, but not different species assemblages. Similar to previous studies, several abiotic factors did not explain patterns in distribution and abundance among beds. The proportion of introduced species was very high (>90% of all individuals), a finding unique among seagrass epifaunal studies to date. Defining numerical patterns in epifaunal communities will inform related efforts to understand effects of epifaunal species and assemblages on eelgrass growth dynamics, seed production, and higher order trophic interactions over space and time.     

Seasonal Analysis of Zoobenthos Associated with a Zostera marina L. Bed in Gulbahce Bay Aegean Sea, Turkey

Abstract. seasonal sampling of three stations in Gulbahce Bay Aegean Sea for the zoobenthic organisms associated with a Zostera marina bed was carried out during 1993–94. Temperature, salinity and oxygen were recorded each sampling period. A total of 7 taxonomic groups were determined: Polychaeta was the dominant group comprising 77 % of the total taxa and individuals, followed by Crustacea, Bivalvia and other groups such as Nemertea. Sipuncula, Turbellaria and Phoronida. Of the 108 taxa encountered, the polychaetes Notomastus latericeus and Caulleriella alata accounted for 49 % of the total populations. The species abundance did not show major changes among seasons, with highest values in fall 982 indiv. ·m^-2 and lowest in spring 754 indiv. ·m^-2. Diversity and evenness of the samples were relatively high and fairly constant, and were affected by the dominance levels of the species.