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DNA barcode assessment of Ceramiales (Rhodophyta) in the intertidal zone of the northwestern Yellow Sea

A total of 142 specimens of Ceramiales(Rhodophyta) were collected each month from October 2011 to November 2012 in the intertidal zone of the northwestern Yellow Sea. These specimens covered 21 species,14 genera,and four families. Cluster analyses show that the specimens had a high diversity for the three DNA markers,namely,partial large subunit r RNA gene(LSU),universal plastid amplicon(UPA),and partial mitochondrial cytochrome c oxidase subunit I gene(COI). No intraspecific divergence was found in our collection for these markers,except for a 1–3 bp divergence in the COI of Ceramium kondoi,Symphyocladia latiuscula,and Neosiphonia japonica. Because short DNA markers were used,the phylogenetic relationships of higher taxonomic levels were hard to evaluate with poor branch support. More than half species of our collection failed to find their matched sequences owing to shortage information of DNA barcodes for macroalgae in Gen Bank or BOLD(Barcode of Life Data) Systems. Three specimens were presumed as H eterosiphonia crispella by cluster analyses on DNA barcodes assisted by morphological identification,which was the first record in the investigated area,implying that it might be a cryptic or invasive species in the coastal area of northwestern Yellow Sea. In the neighbor-joining trees of all three DNA markers,H eterosiphonia japonica converged with D asya spp. and was distant from the other Heterosiphonia spp.,implying that H. japonica had affinities to the genus Dasya. The LSU and UPA markers amplified and sequenced easier than the COI marker across the Ceramiales species,but the COI had a higher ability to discriminate between species.     

Molecular markers for identifying a new selected variety of Pacific white shrimp Litopenaeus vannamei

Selective breeding of the Pacific white shrimp Litopenaeus vannamei during the last decade has produced new varieties exhibiting high growth rates and disease resistance.However,the identification of new varieties of shrimps from their phenotypic characters is difficult.This study introduces a new approach for identifying varieties of shrimps using molecular markers of microsatellites and mitochondrial control region sequences.The method was employed to identify a new selected variety,Kehai No.1(KH-1),from three representative stocks(control group):Zhengda;Tongwei;and a stock collected from Fujian Province,which is now cultured in mainland China.By pooled genotyping of KH-1 and the control group,five microsatellites showing differences between KH-1 and the control group were screened out.Individual genotyping data confirmed the results from pooled genotyping.The genotyping data for the five microsatellites were applied to the assignment analysis of the KH-1 group and the control group using the partial Bayesian assignment method in GENECLASS2.By sequencing the mitochondrial control regions of individuals from the KH-1 and control group,four haplotypes were observed in the KH-1 group,whereas14 haplotypes were obtained in the control group.By combining the microsatellite assignment analysis with mitochondrial control region analysis,the average accuracy of identification of individuals in the KH-1group and control group reached 89%.The five selected microsatellite loci and mitochondrial control region sequences were highly polymorphic and could be used to distinguish new selected varieties of L.vannamei from other populations cultured in China.     

香鱼(Plecoglossus altivelis)肿瘤坏死因子-α (TNF-α) 基因的分子克隆、鉴定及免疫相关性表达

肿瘤坏死因子-α(tumor necrosis factor-alpha, TNF-α)是TNF 超家族成员之一, 在动物机体抵 抗细菌和病毒入侵时发挥重要作用。本研究基于香鱼(Plecoglossus altivelis)单核/巨噬细胞转录组数 据库, 获得了香鱼TNF-α 基因(PaTNF-α)全长cDNA 序列。PaTNF-α 由1932 个核苷酸组成, 包含一 个大的开放阅读框, 编码235 个氨基酸, 预测分子量为26.4 kDa.氨基酸序列多重比对分析表明, PaTNF-α 具有TNF 超家族的特征序列、1 个TACE 酶切位点和1 个保守的二硫键, 与虹鳟 (Oncorhynchus mykiss)TNF-α 同源性最高, 为53.4%; 系统进化树分析表明, 鱼类TNF-α 形成一个大 簇, PaTNF-α 与虹鳟TNF-α 进化相关性最高。实时荧光定量PCR (quantitative real-time PCR, qRT-PCR) 结果显示, PaTNF-α mRNA 在头肾中表达量最高; 鳗弧菌(Vibrio anguillarum)感染后香鱼肝、脾、头 肾和外周血细胞中PaTNF-α mRNA 表达量显著上调; 香鱼单核/巨噬细胞经鳗弧菌、LPS 和poly(I: C) 处理后, PaTNF-α mRNA 表达量显著上调。原核表达了PaTNF-α 成熟肽, 并制备了抗血清。Western blot 分析表明, 鳗弧菌感染后香鱼血清和单核/巨噬细胞上清中的PaTNF-α 表达量含量也显著增加。 综上, 香鱼TNF-α mRNA 及蛋白的表达与病原体感染密切相关, 为深入研究鱼类TNF-α 的生物学功 能及其在病原体感染中的作用机制提供理论基础。     

未来黄、东海营养盐浓度变化情景预测

本文基于FGOALS(Flexible Global Ocean-Atmosphere-Land System model)对未来气候情景的预测结果,结合千年生态评估的未来两个情景下的河流营养盐载荷特征,利用黄、东海水动力模型和生态模型并采用降尺度的方法对未来黄、东海营养盐的分布特征进行情景预测。结果表明,两个情景下未来河口邻近海区营养盐浓度将显著增加,富营养化加剧;GO(Global Orchestration)情景下,河流无机氮载荷增幅较大,夏季黄海中部无机氮浓度明显升高;AM(Adapting mosaic)情景下,由于河流无机磷载荷增幅较大,海区氮磷比有所下降,夏季黄海中部表层无机氮浓度降低,而在底层升高。通过敏感性实验并结合收支分析对各海区水动力条件未来变化、河流载荷变化的相对贡献进行了评估:相对于水温和水动力环境改变,河流营养盐排放量的增长是未来营养盐浓度增加的主要原因。营养盐收支分析表明,未来对流和混合输运的变化有助于黄海营养盐浓度的增加,夏季生物量升高造成更多碎屑沉降并在底层矿化使得层化季节冷水团底部营养盐浓度增长;长江口邻近海区营养盐浓度增长主要受冲淡水羽流的影响;净初级生产增加加剧了营养盐的消耗。     

便携式高频地波雷达台湾海峡浪高观测

As an important equipment for sea state remote sensing, high frequency surface wave radar(HFSWR) has received more and more attention. The conventional method for wave height inversion is based on the ratio of the integration of the second-order spectral continuum to that of the first-order region, where the strong external noise and the incorrect delineation of the first- and second-order Doppler spectral regions due to spectral aliasing are two major sources of errors in the wave height. To account for these factors, two more indices are introduced to the wave height estimation, i.e., the ratio of the maximum power of the second-order continuum to that of the Bragg spectral region(RSCB) and the ratio of the power of the second harmonic peak to that of the Bragg peak(RSHB). Both indices also have a strong correlation with the underlying wave height. On the basis of all these indices an empirical model is proposed to estimate the wave height. This method has been used in a three-months long experiment of the ocean state measuring and analyzing radar, type S(OSMAR-S), which is a portable HFSWR with compact cross-loop/monopole receive antennas developed by Wuhan University since 2006. During the experiment in the Taiwan Strait, the significant wave height varied from 0 to 5 m. The significant wave heights estimated by the OSMAR-S correlate well with the data provided by the Oceanweather Inc. for comparison, with a correlation coefficient of 0.74 and a root mean square error(RMSE) of 0.77 m. The proposed method has made an effective improvement to the wave height estimation and thus a further step toward operational use of the OSMAR-S in the wave height extraction.     

应用实时定量PCR技术检测秦皇岛近岸海域褐潮原因种——抑食金球藻(Aureococcus anophagefferens)

Aureococcus anophagefferens, a small pelagophyte algae, has caused brown tide blooms in coastal waters of Qinhuangdao in recent years, presenting significant negative impacts on the shellfish mariculture industry. Under standard light microscopy, it is visually indistinguishable from other small algae in field samples due to its extremely small size. In this study, quantitative polymerase chain reaction(q PCR) based on 18 S r DNA sequences was developed and used to detect and enumerate A. anophagefferens. A linear regression(R2 = 0.91) was generated based on cycle thresholds value(Ct) versus known concentrations of A. anophagefferens. Twenty-two field samples collected in coastal waters of Qinhuangdao were subjected to DNA extraction and then analyzed using q PCR. Results showed that A. anophagefferens had a wide distribution in coastal waters along Qinhuangdao. Elevated A. anophagefferens abundance, category 3 brown tide blooms(200 000 cells/m L) occurred at Dongshan Beach and Tiger-stone Beach in August in 2013. In shellfish mariculture areas along coastal waters of Qinhuangdao, 4 stations had category 3 blooms, and 6 stations had category 2 blooms(35 000–200 000 cells/m L) in August and all stations had category 1 blooms(0 to ≤35 000 cells/m L) in October. Quantitative PCR allows for detection of A. anophagefferens cells at low levels in filed samples, which is essential to effective management and prediction of brown tide blooms.     

新型张力腿式储油处理平台的动力响应分析

针对目前小油田早期生产中存在的不足,提出一种适用于浅水海域的新型张力腿式储油处理平台,该平台主要由储油箱、浮箱、吸力桩、系泊链和甲板结构构成,具备原油处理和储存功能。应用非线性时域耦合分析法,研究了有义波高、平均波周期、水深对该平台的运动响应和系泊链张力的影响。计算结果表明,波高对平台动力响应有较大影响,有义波高每增加0.4 m,平台水平位移相应增加0.408 m,而系泊链力随有义波高的增加呈准线性增大;波周期对平台水平运动影响显著,平均波周期从6.5 s逐级变化至8.5 s时,最大水平位移自3.760 m渐增至5.467 m,而波周期对系泊链力影响较小,变化率一般小于10%;该新型平台对水深大于20 m的浅水海域有良好的动力特性,而对水深小于20 m的海域却表现异常。因此,该新型张力腿式平台能满足水深不小于20 m浅海区的油田早期生产需要。     

一般地形大光斑激光测高回波模拟研究

国内对基于陆地模型的地表回波仿真的研究较多,主要分析了三种特定的地表模型:平面、阶梯、斜坡地形,取得了不错的进展,但是却不具有一般性。针对这种情况,本文从一般性地表模型出发,根据实际的地形三维离散点,用Delaunay三角形剖分拟合出地表模型,用剖分后的三角面片返回的能量代表整个地表返回的能量,拟合出回波波形。通过和GLAS系统获取的相同区域的实际回波波形进行相关性比较,结果发现两者具有很好的相关性,验证了算法的正确性。文章结论为后续继续深入开展激光大光斑内精细地形信息分析、激光测高数据与光学影像复合处理提供了理论基础,为发展我国自主的星载激光测高卫星的有关指标论证提供参考。