中国海洋渔业转型与深蓝渔业战略性新兴产业

中国的渔业发展为保障粮食安全、增加优质动物蛋白供应发挥了重要作用。由于近海渔业资源衰退、水域环境恶化、生产方式粗放、产业结构不合理等影响因素,海洋渔业发展空间受到了限制,拓展渔业发展空间,向深远海域发展的需求与日俱增。该文从中国渔业转型的发展要求、发展现状和制约因素出发,重点分析了深远海养殖的发展潜力,提出了深蓝渔业“养捕加一体化”的概念。阐明了深蓝渔业由深远海工业化养殖、海洋生物战略性种群资源利用、水产品高值化加工与冷链物流系统、种质资源开发和渔业船联网等要素构成,具体阐述了其发展方式,旨在为推进中国深蓝渔业发展提供参考。

养殖金乌贼卵子发生和卵巢发育的组织学观察

为了深入了解金乌贼(Sepia esculenta Hoyle)的生殖生理特性,本研究采用解剖学方法及组织切片技术,观察和描述了金乌贼卵巢形态和卵子各个时相结构特征及其变化规律。研究结果表明,金乌贼卵子发生过程的主要特征表现为,卵子成熟之前滤泡细胞一直存在且伴随着其结构变化,滤泡细胞解体与卵黄物质出现同步发生,卵黄物质从无到有再到大量积累直至充满整个卵母细胞,细胞核则随着滤泡细胞内折逐渐发生偏移直至最后消失。研究还发现金乌贼卵巢内卵子发生不同步且形态差异显著,未成熟的卵子粘连在卵巢基质上,依次环布于卵巢基质周围,越靠近中心,卵母细胞发育程度越低;成熟后的卵子游离在卵巢腔中,经过输卵管排出体外。根据卵巢内卵子的大小和形态、滤泡细胞形态及卵黄形成情况,将金乌贼卵子发生分为增殖期(卵原细胞期)、小生长期(无滤泡期,单层滤泡期,双层滤泡期)、大生长期(滤泡内折期,卵黄形成早期,卵黄形成晚期)、成熟期、退化吸收期共5个时期9个时相。卵巢发育分期则是以卵子时相为基础,根据卵巢的外观形态、性腺系数以及切面上优势卵子的时相,把卵巢分为Ⅰ~Ⅵ共6个时期。金乌贼卵巢发育和卵子发生的动态观察与描述,对丰富头足类繁殖生物学理论及提升人工繁殖技术都具有重要意义。

魁蚶C型凝集素基因cDNA的克隆及表达分析

通过cDNA末端快速扩增(Rapid amplification of c DNA ends,RACE)技术克隆得到魁蚶(Scapharca broughtonii)C型凝集素(C-type lectin,Sb-Lec1)基因,该基因全长为700 bp,其中,5′-UTR为29 bp,3′-UTR为167 bp,开放阅读框长度为504 bp,编码167个氨基酸,包括长度为23个氨基酸的信号肽序列、129个氨基酸的糖识别结构域(CRD)以及参与二硫键形成的6个半胱氨酸。预测蛋白分子量为19.11 k Da,理论等电点为4.74。多序列比对结果显示,Sb-Lec1基因CRD编码的氨基酸序列与长牡蛎(Crassostrea gigas)、紫贻贝(Mytilus galloprovincialis)和海湾扇贝(Argopecten irradians)C型凝集素的同源性分别为38%~40%、34%~35%和38%~39%,Sb-Lec1基因编码的氨基酸序列与其他物种的凝集素基因具有相似的结构,均含有形成二硫键的4个保守半胱氨酸。系统进化分析结果显示,魁蚶先与贝类聚为一支,再与脊椎动物聚在一起,表明魁蚶Sb-Lec1基因在进化树上的位置与其传统分类所处位置一致。采用荧光定量PCR技术,检测了Sb-Lec1基因在组织中的表达情况,发现其在肝胰腺、血淋巴、鳃、外套膜、闭壳肌、斧足中均有表达,其中,肝胰腺表达量最高。同时,分析了Sb-Lec1基因在鳗弧菌(Vibrio anguillarum)刺激下的mRNA表达量变化情况。结果显示,与对照组相比,菌刺激组Sb-Lec1基因mRNA在各检测组织中的表达量均显著上调(P<0.05),随着刺激时间的延长,表达量呈先升高后降低的趋势。本研究表明,魁蚶Sb-Lec1基因在机体免疫防御方面发挥重要功能。

金乌贼生长期DNA甲基化水平和模式

DNA甲基化在基因表达、细胞衰老、性状分化中发挥重要调控作用,为探讨生长期金乌贼(Sepia esculenta Hoyle)不同性别、不同组织DNA甲基化水平和模式的差异,本研究采用荧光标记的甲基化敏感扩增多态性(Fluorescence-labeled Methylation Sensitive Amplified Polymorphism,F-MSAP)技术,选取12对特异性引物,检测分析了雌、雄金乌贼肌肉、心脏、胰脏和性腺4种组织的基因组DNA甲基化。结果显示,生长期金乌贼基因组DNA总甲基化水平为23.97%~39.70%,在水产无脊椎动物中处于较高水平;金乌贼4种组织中,肌肉的总甲基化水平最高,这可能与金乌贼存在异速生长现象且在生长期运动器官优先发育有关;金乌贼甲基化水平和模式存在性别差异,雌性金乌贼肌肉组织DNA总甲基化水平显著低于雄性,心脏和胰脏组织DNA总甲基化水平却显著高于雄性;此外,雌性金乌贼肌肉组织全甲基化水平对总甲基化水平贡献最大,与之不同的是,雌性其他组织、雄性金乌贼各组织中半甲基化水平和全甲基化水平差异不大,说明金乌贼DNA甲基化的水平和模式具有性别和组织差异。上述结果可为深入研究金乌贼生长发育、组织分化和衰老死亡等生命过程的表观遗传学调控提供基础数据。

金乌贼端粒酶逆转录酶基因鉴定和组织差异表达分析

本研究从金乌贼(Sepia esculenta)转录组中筛选出疑似TERT(Telomerase reverse transcriptase)基因序列,通过系统进化和同源序列分析,鉴定出该基因为金乌贼TERT基因,序列长度为2415 bp。采用实时荧光定量PCR技术,检测和分析金乌贼TERT基因在不同组织、不同发育时期和不同性别的相对表达量差异。结果显示,与产卵期相比,濒死期的心、肝、胰、鳃、性腺、食道上神经团(除去视腺和视叶后的脑组织)等6个组织的TERT基因表达量呈显著下调趋势,推测与濒死期的器官组织分裂和增殖能力下降有关。TERT基因在金乌贼不同组织中具有明显的组织特异性,说明金乌贼各组织器官的衰老状态并不一致。本研究结果可为深入研究头足类生殖与衰老等生命过程的端粒酶调控机制提供基础数据。

RAPD analysis of wild stock of penaeid shrimp (Penaeus chinensis )in Chinese coastal waters of the Huanghai Sea and coastal waters of the Bohai Sea

Genetic diversity of 32 individuals of P. chinensis in the Chinese coastal waters of the Huanghai and Bobal Seas was detected by RAPD technique. Twenty decamer primers of OPI were used for DNA amplification for each individual. The results showed that: Seventeen primers obtained reproducible fingerprints, and the bands were clear. Thirty--nine of 106 loci detected were polymorphic, amounting to 36. 8 %. Mean genetic distance was 0. 094 1 0. 020 6; 68 markers (63. 2 % of the total) showed stable homogeneity in all of the 32 individuals.

Evaluation of reference genes for quantitative real-time PCR analysis of gene expression during early development processes of the tongue sole(Cynoglossus semilaevis)

Differential expression of genes is crucial to growth and development of fish. To select the appropriate genes for gene normalization during Cynoglossus semilaevis early developmental process, eight candidate reference genes （ACTB, B2M, EF1A, GADPH, RPL7, TUBA, UBCE and 18S） were tested for their adequacy by using quantitative real-time PCR. The results showed that the expression of all the examined genes exhibited tissue dependent variations in the mature C. semilaevis. EFIA was listed as the most stable reference among the 14 tissues by RefFinder. Furthermore, the recommended comprehensive ranking of the stability determined by RefFinder showed that 18S was the most stable gene during the early developmental stages （from oosphere to 90 days old） in this study. However, when divided the Ct value data of the above mentioned early developmental stages into two separate periods （embryo and post-hatching periods）, TUBA and 18S represented the most stable references of these two developmental periods, respectively. Consequently, the reference gene should be carefully and accurately chosen even for studies of the same species at various developmental processes. The relevant data may help in selecting appropriate reference genes for mRNA expression analysis, and is of great value in the studies of fish growth and development.

Expression profiles of sex-related genes in gonads of genetic male Takifugu rubripes after 17β-estradiol immersion

Estradiol treatment during early life stages of tiger puffer Takifugu rubripes induces feminization in genetic males.However,the ovaries in genetic males may revert to testes once estradiol treatment is halted.Therefore studies should investigate molecular mechanisms underlying ovary-to-testis recovery in genetic males after treatment.In the present study,tiger puffer were exposed to 10,and 100μg/L 17β-estradiol(E 2)from 15 to 100 days post-hatching(dph),then gonad phenotypes and expression profi les of six sex-related genes(cyp19a,foxl2,dmrt1,amh,sox9a,and sox9b)were characterized after the exposure.Results showed that both 10 and 100μg/L E2 induced ovarian development in genetic males at 100 dph.However,all ovaries induced by 10μg/L E2 first developed into intersexual gonads and subsequently reverted to testes after the exposure.As for treatment of 100μg/L E2,while the rest of the ovaries maintained morphological stability,percentages of intersexual gonads reached 38%-57%,and none were reverted to testes.Increased mRNA levels of cyp19a,foxl2 and sox9b and decreased mRNA levels of dmrt1,amh,and sox9a were observed during the ovarian development in genetic males.While contrary gene expression profiles were detected during ovary-to-testis transformation.The mRNA levels of all the six genes were increased during the development of intersexual gonads.These results indicated that up-regulation of dmrt1,amh and sox9a is associated with initial ovary-to-intersexual transformation,and suppression of foxl2,cyp19a and sox9b is essential for complete ovary-to-testis recovery in genetic males.This research will help to trace the molecular processes underlying gonadal transformation in teleosts.

Genetic Variation Among White Croaker Populations

To investigate the genetic structures and differentiation of different wild populations of white croaker (Pennahia argen- tata), horizontal starch gel electrophoresis was performed on 133 individuals collected from five different locations in China and Japan. The eleven enzyme systems revealed 15 loci, of which eleven were polymorphic. The percentage of polymorphic loci of white croaker populations varied from 6.67% to 53.33%; the mean observed and expected heterozygosity ranged from 0.0033 to 0.0133 and 0.0032 to 0.0191, respectively. The expected heterozygosity revealed a low genetic variability for white croaker in comparison with other marine fishes. The genetic distances between populations ranged from 0.00005 to 0.00026. A weak differentiation was observed within each clade and between clades; and no significant differences in gene frequencies among populations were observed in white croaker. Among the five populations, three Chinese populations showed more genetic diversity than that in Japanese populations.

The point of no return and pectoral angle of Japanese anchovy(Engraulis japonicus) larvae during growth and starvation

At a temperature of 23.0 - 24.8℃, the mixed feeding of Japanese anchovy larvae was initiated 24 h before the yolk-sac was exhausted. The point of no return （PNR） was reached on the 6th day after hatching. On the 4th day after hatching, the pectoral angle appeared in both fed and unfed anchovy larvae although it was more evident and sharper in the starved and the PNR stage larvae than in the fed ones. According to observations of larvae collected in the sea, the pectoral angles were evident not only in the larvae of 3.62 - 7.44 mm in standard length, but also in the larvae of 2.70 mm in standard length with remnants of yolk. The pectoral angles became diffuse when the larvae reached 7.84 mm and vanished at 9.86 mm. The pectoral angle cannot be used as a criterion to distinguish healthy from starving larvae.

Corrigendum to: Expression profiles of sex-related genes in gonads of genetic male Takifugu rubripes after 17β-estradiol immersion

The original version of this article unfortunately contained a mistake. The publication year in the header on the first page (p.1113) of this article was incorrect. The corrected publication year is given below:

Cloning, expression profiling and promoter functional analysis of bone morphogenetic protein 2 in the tongue sole(Cynoglossus semilaevis)

BMP2 plays crucial roles in vertebrate developmental process and acts as a bone inducer during osteogenesis. We present here the molecular cloning of bmp2 cDNA from the marine flatfish Cynoglossus semilaevis, and the analysis of bmp2 expression profiling and promoter function. The full length of bmp2 cDNA sequence is 2 048 bp,which encodes a protein of 422 amino acids. Tissue expression distribution of bmp2 was examined in 14 tissues of mature individuals by quantitative real time PCR（qRT-PCR）. The results revealed that bmp2 was expressed ubiquitously, and the highest expression level was detected in the spinal cord. Moreover, bmp2 expression levels were detected at 15 sampling time points of early developmental stages（egg, larva, juvenile and fingerling stages）.The highest expression level of bmp2 was observed at the gastrula stage, which was about ten times higher than those at the other three embryo stages. Whole-mount in situ hybridization showed that the bmp2 signal was strongly detected at the location of the crown-like larval fin, heart and liver, and slightly expressed in the notochord at one day post hatch（dph）; then the expression of bmp2 started to be concentrated in notochord at three dph. Subsequently, we characterized the 5′-flanking region of bmp2 by testing the promoter activity by Luciferase reporter assays. Positive regulatory region was detected at the location of –179 to ＋109. The predicted transcription factor binding sites（E-box binding factors, zinc finger transcription factor, etc.） in this region might participate in the transcriptional regulation of the bmp2 gene.

中国近海带鱼Trichiurus japonicus的命名和分类学地位研究

带鱼属（Trichiurus）鱼类是中国最重要的海洋渔业捕捞对象之一。然而,有关中国南北沿海均有分布的这种带鱼（简称中国近海带鱼）应取何名、属于何种及其系统进化过程等问题仍未有统一的认识和定论。本研究通过测定中国近海带鱼14个群体共计166个个体的线粒体16S r RNA基因序列,分析该物种与中国近海其他3种带鱼属鱼类的序列变异,比较该种带鱼与世界其他带鱼属有效种的形态差异,综合探讨其命名、分类地位及系统进化过程。基于Kimura双参数模型计算,中国近海带鱼（Tsp-CH）与台湾（Tj-WT）及日本（Tj-JP）的带鱼（Trichiurus japonicus）的组间净遗传距离（0.000 7-0.002 1）远低于带鱼属内其他种间的净遗传距离（0.065 6-0.082 0）,也明显低于高鳍带鱼（Trichiurus lepturus）地理群体间的遗传距离（0.010-0.016）;分子变异分析（AMOVA）和FST分析均表明这三组带鱼组间遗传分化不显著。结果表明,中国近海带鱼与来自台湾及日本的带鱼（T.japonicus）的遗传分化处于种内水平。采用邻接法构建的分子系统树显示,不同地理来源的带鱼（T.japonicus）（即包括Tsp-CH,Tj-WT,Tj-JP）与高鳍带鱼（T.lepturus）、南海带鱼（Trichiurus nanhaiensis）、短带鱼（Trichiurus brevis）等3种带鱼属鱼类互为单系,在属内各自形成独立的进化分支。结合分子数据和形态学比较研究资料,本研究确认中国南北沿海均有分布的这种带鱼是Trichiurus japonicus Temminck et Schlegel,1844,它在种的分化、形成、分布及形态上与世界其他带鱼属鱼类存在系列差异。