基于GBS技术的黄渤海沿岸托氏琩螺遗传特征分析

为了探讨辽宁沿海非经济贝类野生种群的遗传多样性水平和遗传分化特征,本研究选取了托氏琩螺(Umbonium thomasi),利用基于简化基因组测序的基因分型技术(GBS)对辽宁黄渤海沿岸的5个群体进行了全基因组水平的SNP标记开发和遗传特征分析。本研究共获得1315987个SNP标记,发现这5个托氏琩螺群体的观测杂合度和期望杂合度分别为0.0851～0.1161和0.1424～0.1627,观测杂合度均低于期望杂合度,说明托氏琩螺群体存在杂合子缺失,有种群退化风险。C4-ZH群体可能因为外来物种的入侵而发生种群缩小和多样性降低。遗传分化研究发现, 5个托氏琩螺群体处于中等分化水平,其中C5-JZ群体因为区域保护原因与其他4个群体的分化程度相对较高,且通过群体遗传结构分析发现,C5-JZ群体与其他群体的基因交流较少,基于此本研究开发了3个C5-JZ群体的特征性SNP标记。本研究结果将有助于了解辽宁沿海非经济物种的多样性水平,为自然资源的保护和利用提供依据。     

额尔齐斯河银鲫形态学及COI基因序列分析

采用传统形态学方法和线粒体COI基因序列分析对额尔齐斯河银鲫(Carassius auratus gibelio)两个群体的形态和遗传差异进行了比较。结果显示:通过对8个可量性状和20个框架数据的主成分分析可以看出,两个群体分别占据两个不相重叠的区域,两个群体在形态上存在明显分化。COI基因序列分析显示,两群体之间遗传距离为0.012,群体间遗传距离是群体内最大遗传距离的4倍,两群体间在编码的氨基酸序列上出现了一处替换,群体遗分化指数值为0.96078,基因流值为0.01,表明群体间缺乏基因交流,群体间有明显的遗传分化。     

Gonadal morphogenesis and sex differentiation in cultured chub mackerel,Scomber japonicus

The timing of primordial germ‐cell (PGC) migration with regard to the gonadal anlagen, gonad formation and sex differentiation was examined histologically in the chub mackerel (Scomber japonicus) at 5–190 days post hatching (dph). At 5 dph, PGCs appeared on the peritoneal epithelium surface or in the mesentery, on the dorsal side of the abdominal cavity. By 10 dph, stromal cells around the PGCs proliferated. The gonadal primordium was formed by 15 dph. The gonadosomatic index was 0.01% at 30 dph and increased thereafter (0.32% in females and 0.04% in males at 160 dph). Ovarian differentiation occurred at 30–40 dph, indicated by ovarian cavity formation (elongation and fusion of the upper and lower ovarian edges). Meiosis was subsequently initiated. A few meiotic oocytes surrounded the cavity at 50 dph; most were in the perinucleolus stage at 60 dph and attained a diameter of 60–70 μm at 190 dph. Testicular differentiation occurred at 30 dph, indicated by the formation of the sperm duct primordium. Spermatogonia gradually proliferated, developing into spermatocytes at the chromatin–nucleolus stage (after 90 dph) and subsequently into spermatids and spermatozoa (160 dph). These data could aid the development of seeding and cell‐engineering technologies for scombrid fish.     

海南“石斑鱼苗网”渔具属性辨析

对在2004年7~8月间南海区小型海洋渔具渔法调查中发现的“石斑鱼苗网”的渔具结构、属性、性能和管理进行了分析讨论,研究认为“石斑鱼苗网”属于敷网类,网团型,延绳式。对石斑鱼苗有良好的诱捕效果,从保护资源的角度出发,应对其实施规范管理。     

Differentiation and development of Leydig cell, and induction of spermatogenesis during testicular differentiation in the Japanese eel, Anguilla japonica

The initial appearance and the development of Leydig cells (LCs), the sites of steroid hormone production in the testis, were investigated ultrastructurally during testicular differentiation in the Japanese eel, Anguilla japonica. In addition, the effects of a single injection of human chorionic gonadotropin (HCG; 5 IU g body weight^-1) on histological changes of the testes and serum 11-ketotestosterone (11-KT) were examined at various stages (15–18, 20–23, 26–29, 32–35, 38–41 and 46–50 cm body length (BL)) of testicular differentiation. Testicular differentiation was morphologically characterized by the development of loose connective tissue on the medial side in animals 18–29 cm in BL. Ultrastructurally, LCs were first identified in the loose connective tissue of the testis of the 23 cm fish. In the testes of fish over 32 cm, clusters of LCs were distributed throughout the interstitial region accompanying the increase in number of spermatogonia. In fish larger than 32 cm, spermatogenesis was induced by administration of HCG; serum 11-KT levels were also raised. On the other hand, there was no effect on spermatogenesis or serum 11-KT levels in fish less than 29 cm, or in the controls. These result suggests that morphological differentiation of LCs occurs in testis of the 23 cm eel, and subsequently, the testes of eels of BL more than 32 cm acquire the capability to produce steroid hormones.     

胚胎干细胞的研究进展及应用前景

干细胞(Stem Cell)是一类具有分化潜能和自我复制的早期未分化细胞。胚胎干细胞(Embryonic stem cells,ES细胞)是一种早期胚胎内细胞团(inner cell mass,ICM)或原始生殖细胞(primordial germ cell,PGC)经体外分化抑制培养,分离和克隆得到的具有发育全能性的高度未分化细胞。本文综述了胚胎干细胞的形态学、生长特性、免疫学鉴定方法,ES细胞抑制分化和诱导分化的机理以及饲养层、血清和细胞因子等影响胚胎干细胞分离克隆的因素,并进一步阐述了胚胎干细胞的应用前景以及存在的问题。     

基于牡蛎疱疹病毒DNA聚合酶基因的巢式PCR检测方法的建立及应用

为了建立适用于Os HV-1不同变异株的检测方法,在牡蛎疱疹病毒(Os HV-1)3个变异株全基因组序列比对的基础上,筛选到牡蛎疱疹病毒基因组中高度保守的DNA聚合酶(DNA polymerase)基因,据此设计巢式PCR引物,优化PCR反应体系和条件,建立了基于Os HV-1 DNA聚合酶基因的巢氏PCR检测方法(P-n PCR检测方法),利用P-n PCR与Cn PCR检测方法对不同年份和宿主来源的Os HV-1疑似感染样本进行检测。结果显示,Pn PCR检测方法能稳定地检出100拷贝/μL的病毒DNA;P-n PCR较C-n PCR检测方法具有更强的特异性和更高的检出率。研究表明,本研究建立的P-n PCR检测方法适用于Os HV-1不同变异株的检测,可为该病毒的检测和流行病学调查提供可靠的技术支持。     

西伯利亚鲟性腺早期发生、分化、发育的组织学观察

利用组织学方法研究人工繁育、养殖的西伯利亚鲟(Acipenser baerii)仔、幼、成鱼性腺发育过程中原始生殖细胞(Primordial germ cells,PGC)的迁移,生殖褶的生成,性腺雌雄分化和发育过程。结果表明,西伯利亚鲟于3dph(Days post-hatch)时,PGC以单细胞的形式存在于仔鱼卵黄囊上方两中肾管之间,10dph时其迁移到背侧的腹膜上皮上,13dph时其沿腹膜上皮朝肾管区下方迁移,22dph时生殖嵴形成,31dph时原始性腺形成,40～60dph时原始性腺发育形成裂腔,PGCs沿腔壁排列,裂腔经过进一步发育并从背离腹膜上皮的底端慢慢愈合成实体,90dph时原始性腺中出现生殖上皮和性细胞,150dph时脂肪细胞和血管增多,210dph时性腺出现2种不同的表面结构,性腺开始分化。本研究利用组织学方法借鉴鱼类性腺分期标准,将西伯利亚鲟性腺分化划分为0期和Ⅰ～Ⅵ期,并描述了各期精巢、卵巢的结构变化以及细胞形态学特征。     

‘国光’苹果及其红色芽变花青苷合成与相关酶活性的研究

 以‘国光’苹果及其红色芽变为试材, 测定了果实发育期间的花青苷含量及其相关酶活性,并研究了套袋对芽变花青苷合成的影响。结果显示: ①在果实发育期间, 芽变果皮的花青苷含量明显高于‘国光’, 尤其是成熟期芽变果皮花青苷含量为132170 U·g^-1FW, 而‘国光’仅为49140 U·g^-1FW; ②在果实发育期间, 两个品种间PAL 和UFGT的酶活性无明显差异, 但芽变的CHI和DFR酶活性明显高于‘国光’, 表明芽变花青苷合成能力的提高与CHI和DFR酶活性高有关; ③套袋抑制芽变果皮花青苷的合成, 但解袋后花青苷的含量极显著升高, 解袋后4种酶的变化趋势差异较大, CHI和UFGT活性均迅速升高, 明显高于对照, 这与解袋后花青苷迅速合成相吻合。综上结果, 芽变与原有品种在着色机理上的关键指标是果皮花青苷含量和CHI酶活性。