微囊藻群体总RNA提取方法的比较

微囊藻群体富含多糖类物质是影响微囊藻RNA提取的关键因素.为了获得高质量的微囊藻群体总RNA,对比分析4种针对多糖含量较高的方法——方法 1 PGTX-bead法、方法 2 CTAB-bead法、方法 3 Fast RNAPro Blue Kit和方法 4RNeasy Mini Kit对微囊藻群体总RNA的提取效果.采用琼脂糖凝胶电泳检测微囊藻群体RNA的完整性,Nanodrop ND1000分光光度计检测RNA纯度及浓度,并采用q PCR检测DNA污染情况.结果表明,4种方法都能从微囊藻群体中提取获得RNA,并在去除DNA后都可以进行RT-PCR等后续实验.方法 1 PGTX-bead提取的RNA产量最高,纯度好,DNA污染小,成本低,适合从微囊藻群体中大量提取RNA;方法 2 CTAB-bead提取的RNA样品产量也较高,但DNA污染严重,适合需要同时提取DNA和RNA的样本;方法 3 Fast RNAPro Blue Kit和方法 4 RNeasy Mini Kit提取的RNA产量都较低,但方法 4操作简单,耗时短,所检测目的基因的相对表达量较高,更适合从少量的微囊藻群体中提取总RNA.     

Ficolin在毛蚶抗弧菌感染中的作用

Ficolin是一类含FBG结构域的蛋白,在动物免疫防御中发挥重要作用。无脊椎动物中,Ficolin主要作为模式识别受体和免疫效应分子发挥免疫功能。本研究从毛蚶中克隆获得一条Ficolin基因(命名为SsFic1),该基因编码区序列长度为897 bp,编码298个氨基酸,推测的蛋白含有典型的FBG结构域和Coiled coil结构域,但未发现胶原样结构域。SsFic1的mRNA在毛蚶各组织中广泛分布,其中在肝胰腺和鳃中表达丰度较高,但SsFic1转录产物在卵细胞和四细胞期未检测到,说明该基因不是母源性免疫基因。副溶血弧菌和脂多糖、葡聚糖等免疫刺激物均可诱导SsFic1显著上调表达。利用RNAi技术抑制SsFic1基因表达,酚氧化酶激活因子PAF和补体相关基因(C1qDC、补体C3和Fic3)的表达量及CAT、SOD和PO的酶活力均显著下降,而TEP、C1q2和Fic2等基因的表达显著上调。敲降SsFic1表达后,感染副溶血弧菌后毛蚶血细胞数量和吞噬能力均显著低于对照组,这也是RNAi实验组毛蚶死亡率和血细胞凋亡率显著升高的原因。综上,SsFic1可能通过调控补体通路和酚氧化酶原激活通路参与毛蚶免疫防御反应,这对贝类免疫学研究及抗病新品系的选育具有重要意义。     

中华哲水蚤(Calanus sinicus)个体核酸含量及其对生长状况的指示作用

采用荧光染色法对黄、东海春、秋季中华哲水蚤(Calanus sinicus)个体核酸含量进行了研究,初步探讨了RNA∶DNA比对中华哲水蚤生长状况的指示作用。结果表明,中华哲水蚤在饥饿培养24h后RNA∶DNA比值显著降低;雄性个体RNA含量和RNA∶DNA比值显著低于雌体和C5期幼体;春、秋季(2009年5月和2010年11月)个体RNA含量和RNA∶DNA比值存在显著的空间变化,两者最高值均出现在近岸;两个月份DNA含量相对稳定,在2.5—3.5μg/ind之间;个体RNA含量和RNA:DNA比值与叶绿素之间显著正相关,说明饵料是中华哲水蚤的生长过程中重要的影响因素。     

Application du réseau des neurones artificiels à la prévision des débits horaires: Cas du bassin versant de l’Eure,France

RÉSUMÉ

Les modèles pluie–débit sont fortement utilisés dans la gestion des risques hydrologiques et la prévision des crues. Dans cette étude, nous présentons un modèle pluie–débit pour la prévision des débits horaires basé sur la technique des réseaux de neurones artificiels (RNA). Ce modèle a été développé et appliqué sur le bassin versant de l’Eure au Nord-Ouest de la France afin de dépasser les problèmes dus à la non-linéarité de la relation pluie–débit et à l’imprécision des données collectées. La création de ce modèle a nécessité plusieurs étapes pendant lesquelles nous avons pu déterminer les paramètres du modèle permettant la compréhension de la complexité hydrologique et la production des informations nécessaires à la prévision. Elles ont abouti à un modèle de réseau de neurones artificiels capable d’effectuer, en quelques secondes, des prévisions des crues efficaces jusqu’à un horizon de prévision de 48 h. Ces résultats confirment que les modèles RNA peuvent jouer un rôle important dans le domaine de la prévision car capables de modéliser la non-linéarité des relations pluie–débit rencontrées sur certains bassins hydrologiques.

Editor Z.W. Kundzewicz Associate editor E. Gargouri     

整体原位杂交法研究斑马鱼(Danio rerio)胚胎发育过程中胸苷酸合成酶(TS)的表达

收集不同发育时期的斑马鱼胚胎,制备DIG标记的TS反义RNA探针,采用整体原位杂交方法研究胸苷酸合成酶(TS)基因在斑马鱼胚胎发育各期的时空表达状况.结果表明,在所取样的各个阶段,TS基因均有转录,但其部位不同,中囊胚过渡前后mRNA显现的部位仅存在于受精卵的动物极,10hpf时存在于整个胚胎的外围,至24hpf后集中...     

Variations of backscatter strength along the super slow-spreading Southwest Indian Ridge between 57°E and 70°E

Simrad EM12 backscatter strength data of the Southwest Indian Ridge (SWIR), between 57°E and 70°E, are used to reveal the along-axis segmentation of this super slow-spreading ridge. The backscatter properties of different geologic domains, like bathymetric highs and oblique basins within the rift valley, are characterized using 66 small test sites. We show that backscatter strength is higher on bathymetric swells, corresponding to segment centres, and lower in deep oblique basins corresponding to axial non-transform discontinuities and fracture zones. This contrast between segment centres and discontinuities is produced by both a thicker sediment cover and less frequent volcanic eruptions at segment ends. Using the model of Mitchell (1993), sediments have been estimated to be 2 to 5 m thicker in these areas than at segment centres. The distribution of the seamounts within the rift valley is controlling the long-wavelength variations of the mean backscatter strength calculated along the axis. Lower densities of seamounts and thicker sediments are producing lower and heterogeneous reflectivity levels in the deepest part of the axial valley floor between 61°45′E and 63°45′E. We propose that cooler mantle temperatures inducing construction of fewer volcanoes occur beneath this part of the ridge. The mean backscatter strength along the SWIR axis decreases dramatically toward the Rodrigues Triple Junction suggesting that volcanic production is reduced between 68°20′E and 69°20′E and that the transition from amagmatic tectonic deformation at the triple junction to new seafloor spreading occurs between 69°20′E and 70°E.     

Sediment trapping and bypassing characteristics of a stable tidal inlet at Kaohsiung Harbor, Taiwan

Around the artificially stabilized tidal inlet that connects Kaohsiung Harbor to Taiwan Straight, 203 surficial samples of the sea floor were taken from the nearshore, in the outer harbor, and portions of the inner harbor. The bathymetry of the same area was also surveyed. The sand fraction in each sample was analyzed for the grain-size composition with a custom-built rapid sediment analyzer. A total of twenty-one size-classes were used in the analysis. Three hypothetical sediment sources were assumed to have influenced the spatial grain-size patterns in the study area: the northward and southward littoral drifts, and the sediments exported from the harbor. After reducing the influence of each hypothetical sediment source separately, the data were analyzed using empirical orthogonal (eigen) function (EOF) analysis. The results indicate that the northward long-term littoral drift is the dominant direction of sediment transport in the nearshore of the study area. A conceptual model for four different sediment trapping and bypassing patterns are proposed. (1) Excess bypass (or net outflux): this bypass pattern is characterized by the export of fine-grained sediments (mud and size classes in the very fine sand fraction) from the interior of the harbor. (2) Partial bypass (or partial trapping): this bypass pattern is represented by the size-classes in fine sand fraction. The amount of these grain sizes entering the outer harbor through the inlet is more than the amount exiting at the inlet mouth, resulting in the retention of a portion of these grain sizes. (3) Total bypass (or zero trapping): grain sizes that exhibit this bypass pattern do not come near the mouth of the inlet. These grain sizes include medium and coarse sand fractions. (4) Lag deposits: this group includes the size classes in the very coarse sand fraction, which are largely concentrated in the scour pit immediately seaward of the inlet mouth. In general, surficial sediment grain-size patterns represent a time-averaged response of the substrate to the transport processes over the time scale of at least two seasons. The differential associations of grain-size groups with various topographic features in the study area suggest morpho-textural relationships exist between the sea floor topography and grain size distribution patterns.     

巨蛎属牡蛎DNA序列的比较及鉴定

【目的】了解巨砺属牡蛎的系统关系和种类区分。【方法】对我国沿海分布的7种巨砺属牡蛎(香港牡蛎、有明牡蛎、熊本牡蛎、岩牡蛎、艾氏牡蛎、太平洋牡蛎、葡萄牙牡蛎)的线粒体DNA片段(COI、12S、16S和tRNAs),以及核糖体RNA基因转录间隔子序列(ITS-1、ITS-2和ITS)进行分子系统学研究。【结果】无论种间或种内,tRNAs序列差异最大,亚种内TS1序列差异最大;tRNAs和COI序列较其它序列变异更快,种间变异与种内变异之间有明显的条形码间隙;12S、16S、ITS和ITS2的种间变异与种内变异无重叠及条形码间隙;ITS1的种间变异与种内变异出现重叠。在亚种层面,12S、ITS、ITS1和ITS2的亚种间变异与亚种内变异出现重叠;16S的亚种间变异与亚种内变异无重叠及条形码间隙;tRNAs与COI序列亚种间变异与亚种内变异之间有明显的条形码间隙,有利于区分亚种。【结论】tRNAs和COI序列可用于种及亚种的鉴定。     

ROBUST: The ROle of BUffering capacities in STabilising coastal lagoon ecosystems

“Buffer capacities” has been defined in ecology as a holistic concept (e.g., Integration of Ecosystem Theories: A Pattern, second ed. Kluwer, Dordrecht, 1997, 388pp), but we show that it can also be worked out in mechanistic studies. Our mechanistic approach highlights that “buffering capacities” can be depleted progressively, and, therefore, we make a distinction between current and potential “buffering capacities”. We have applied this concept to understand the limited “local stability” in seagrass ecosystems and their vulnerability towards structural changes into macro-algal dominated communities. We explored the following processes and studied how they confer buffering capacities to the seagrass ecosystem: (i) net autotrophy is persistent in Zostera noltii meadows where plant assimilation acts as a sink for nutrients, this contrasted with the Ulva system that shifted back and forth between net autotrophy and net heterotrophy; (ii) the Z. noltii ecosystem possesses a certain albeit rather limited capacity to modify the balance between nitrogen fixation and denitrification, i.e., it was found that in situ nitrogen fixation always exceeded denitrification; (iii) the nitrogen demand of organoheterotrophic bacteria in the sediment results in nitrogen retention of N in the sediment and hence a buffer against release of nitrogen compounds from sediments, (iv) habitat diversification in seagrass meadows provides shelter for meiofauna and hence buffering against adverse conditions, (v) sedimentary iron provides a buffer against noxious sulfide (note: bacterial sulfide production is enhanced in anoxic sediment niches by increased organic matter loading). On the other hand, in the coastal system we studied, sedimentary iron appears less important as a redox-coupled buffer system against phosphate loading. This is because most inorganic phosphate is bound to calcium rather than to iron. In addition, our studies have highlighted the importance of plant–microbe interactions in the seagrass meadows.