水产生物基因组研究进展与趋势

本文综述了水产生物基因组研究相关技术的发展历程和关键技术的应用。以二代测序技术的出现为分界点,首先回顾了早期水产养殖生物在遗传学和分子生物学方面的研究结果,以及为开展全基因组测序所做的相关基础研究,然后重点介绍了二代测序技术应用于水产生物全基因组测序和经济性状遗传基础解析的研究进展,最后展望了水产生物基因组研究发展趋势。水产生物经济性状遗传机制高度复杂,从全基因组角度阐明其遗传机制仍有很多难题,但基因决定性状是生物学法则之一,探索这一过程的奥秘引人入胜。     

饲料糊精水平对暗纹东方鲀幼鱼生长、消化酶活性和血液生化指标的影响

配制等蛋白质(42%),等脂肪(9%),糊精水平分别为10%、15%、20%、25%、30%的5组饲料,饲养暗纹东方鲀幼鱼[初始体质量为(10.25±0.51)g]60 d,考察饲料中不同糊精水平对其生长、消化酶活性和血液生化指标的影响。结果发现,随着饲料糊精水平的升高,暗纹东方鲀幼鱼的特定生长率和饲料效率先增加后降低,25%组最高,并显著高于10%和15%组。30%组鱼的脏体比和肝体比显著高于10%组,同时其肝脂肪含量也显著高于其他组。10%组的肝糖原和血浆总氨基酸含量显著低于其他组。10%和15%组鱼的肝胰脏和肠道中淀粉酶活性显著低于其他实验组,而其肝胰脏中胰蛋白酶活性显著高于30%组。25%和30%组的血浆总胆固醇和甘油三酯含量显著高于10%和15%组。10%和15%组的血浆尿素氮含量显著高于其他实验组。15%和20%组的血细胞数量显著高于其他实验组。30%组的细胞凋亡率及细胞内活性氧(ROS)和钙离子(Ca2+)含量显著高于其他实验组。饲料糊精水平对暗纹东方鲀幼鱼的血糖含量、血浆谷草转氨酶和谷丙转氨酶活性及肝胰脏和肠道中脂肪酶活性无显著影响。在本实验条件下,暗纹东方鲀幼鱼饲料中糊精的适宜水平为20%～25%。     

暗纹东方鲀mtDNA的分离纯化及其细胞素b基因的分子克隆

利用改进的差速离心法和碱裂解法制备并分离纯化了暗纹东方纯mtDNA,测得分子大小为19．4Kb。用线粒体特异染色法跟踪和监测线粒体提取,结果表明,25000-35000g能获得90％以上的高得率。利用制备纯化的mtDNA,已首次克隆了暗纹东方纯细胞色素b基因(cytb)。     

大黄鱼选育群体与野生群体杂交 F1生长性状研究

利用大黄鱼野生群体与选育F1养殖群体杂交及群体内自繁,获得选育F1（♀）×野生（♂）、野生（♀）×选育F1（♂）、野生（♀）×野生（♂）和选育F1（♀）×选育F1（♂）4个组合的子代,对其卵径、油球径以及2～10月龄全长、体长和体质量进行分析比较。试验结果表明,选育F1（♀）×野生（♂）卵径最大,并与野生（♀）×野生（♂）之间存在显著性差异（P＜0．05）,4个群体油球径差异不显著（P＞0．05）;体质量是大黄鱼群体间杂交主要表现的生长性状,9月龄体质量双亲杂种优势最大为14．48％,野生（♀）×选育F1（♂）体质量杂种优势最大为25．68％;野生（♀）×选育F1（♂）的全长特定生长率为1．92％／d ,体质量为5．60％／d及选育F1（♀）×野生（♂）体长为1．99％／d ,是4个群体中最大特定生长率;拟合体长（L）与体质量（m）关系,选育F1（♀）×野生（♂）为 m＝0．0277L2．8045（r2＝0．9980）,野生（♀）×选育F1（♂）为 m＝0．0210 L2．9287（r2＝0．9993）,野生（♀）×野生（♂）为 m＝0．0236 L2．8592（r2＝0．9916）,选育F1（♀）×选育F1（♂）为 m＝0．0228 L2．8946（r2＝0．9984）。试验结果表明,通过野生群体与选育养殖群体的杂交能提高大黄鱼子代的经济性状。     

赤点石斑鱼与鞍带石斑鱼杂交子一代肌肉营养成分分析与评价

为分析赤点石斑鱼与鞍带石斑鱼杂交子一代的营养组成,参照国家标准,测定了体质量(182.84±29.35) g杂交石斑鱼肌肉的常规营养成分、氨基酸和脂肪酸组成,并对肌肉营养价值进行了评定。试验结果显示,杂交石斑鱼肌肉水分、粗蛋白、粗脂肪和灰分的含量分别为(74.07±0.71)%、(21.52±0.78)%、(4.03±0.15)%和(1.29±0.07)%。肌肉鲜样中测定了17种氨基酸,总量为(19.88±0.15)%;必需氨基酸和鲜味氨基酸总量分别为(8.64±0.13)%和(7.64±0.16)%,必需氨基酸指数为85.19,必需氨基酸组成符合联合国粮农组织/世界卫生组织标准。肌肉鲜样中含有17种脂肪酸,饱和脂肪酸、单不饱和脂肪酸和多不饱和脂肪酸分别占肌肉脂肪酸总量的(27.63±1.15)%、(22.75±1.22)%和(32.59±1.90)%,其中二十二碳六烯酸和二十碳五烯酸占肌肉脂肪酸总量的(19.27±1.27)%。研究表明,赤点石斑鱼与鞍带石斑鱼杂交子一代具有较高营养价值,可作为新品种进行开发。     

铜陵淡水豚国家级自然保护区浮游植物群落特征与水质评价

经济发展改变了铜陵淡水豚国家级自然保护区的水域水生生态环境,为了揭示该保护区浮游植物群落结构特征与水质状况,分别于2011年6月、7月及9月按对4个代表性断面进行浮游植物群落研究,并根据浮游植物群落结构的物种组成,及其结合Shannon-Weaver指数和 Pielou 均匀度指数对水质营养及污染状况进行评价。结果表明：该保护区浮游植物有86种,其中绿藻门最多,34种,占全部浮游植物种类的39.53%;硅藻次之,29种,占总种类的33.73%;蓝藻门14种,占总种类的16.28%。浮游植物总平均密度为9.79×105 ind•L-1,总平均生物量为0.303mg•L-1,调查区域浮游植物平均密度由高到低依次为铜陵沙头南>铜陵沙头北>萝卜洲尾南>萝卜洲尾北;平均生物量由高到低依次为铜陵沙头南>铜陵沙头北>萝卜洲尾北>萝卜洲尾南。铜陵淡水豚自然保护区水质属于富营养型,处于α-中污染状态。     

Effect of atorvastatin on neointimal hyperplasia of venous autografts in rats

AIM: To investigate the effect of atorvastatin on neointimal hyperplasia of autogenous vein graft in rats. METHODS: The model of autogenous vein graft was prepared by transplanting the external jugular vein into aorta in Wistar rats. The rats were divided into three groups: sham operation group, graft control group and graft experimental group. From three days after transplantation, the rats of autograft experimental group were treated by atorvastatin at a dosage of 5 mg·kg-1·d-1. Four weeks after treatment, venous autografts were removed at autopsy and cut into 4 μm sections. Histopathological examination was carried out to analysis the neointimal hyperplasia of grafted veins. Immunohistochemical staining was conducted to evaluate SMα-actin and PCNA expression of neointimal cells in venous autografts. RESULTS: In venous autograft control and experimental groups, SMα-actin-positive smooth muscle cells were proliferated and accumulated excessively in venous autografts, which resulted in significant neointimal formation and vascular lumen narrowing. Neointima quantitative assay revealed that the neointimal hyperplasia of venous autografts was suppressed obviously in graft experimental group, and its neointimal area and NIA/MA ratio of venous autografts were significantly lower than those in graft control group (P<0.01). Immunohistochemical assay indicated that the PCNA labeling index of neointimal cells was significantly lower in graft experimental group than that in graft control group (P<0.01). CONCLUSION: Atorvastatin significantly inhibits the proliferation of neointimal smooth muscle cells and the development of neointimal hyperplasia of venous autografts in rats. Atorvastatin is a powerful inhibitor of restenosis after vascular reconstructive operation with a potential for therapeutic use.     

西瓜新品种美王的选育

美王是以自交系M16为母本,以自交系W-2为父本配制而成的早熟西瓜一代杂种。露地地膜覆盖栽培,果实发育期30 d（天）左右,全生育期97 d（天）左右;日光温室秋冬茬或冬春茬栽培,果实发育期50 d（天）左右,全生育期108 d（天）左右。植株生长势中等,主蔓第6节着生第1雌花,以后每隔 2～3节现1雌花,结果能力强,田间整齐度好。果实圆球形,果皮绿色覆墨绿色规则条带,瓤大红色,剖面均匀一致,中心可溶性固形物含量11.6 %,边部 10.6 %,VC含量84 mg·kg^-1,风味好,品质佳。单瓜质量3.5 kg左右,每667 m²产量3 800 kg左右,适于露地早熟栽培和保护地栽培。     

Prokaryotic expression and identification of a small peptide of osteopontin

AIM: To express osteopontin 13 peptide (OPN 13) in E.coli, and to test the biological activity of the purified products. METHODS: cDNA fragments containing RGD sequences were cloned into prokaryotic expression vector pET-32c(+) including His coding sequence to construct pET-32c-OPN 13 plasmid. E.coli DH5α transformed by pET-32c-OPN 13 plasmid was induced by IPTG at different concentrations for different times to identify the optimal induction condition. Expressed His-OPN 13 fusion protein was purified via Ni-NTA His Bind Resin metal chelation chromatography, and detected by VSMCs adhesion and migration analysis. RESULTS: His-OPN 13 fusion protein was expressed in soluble manner. The fusion proteins were purified via Ni-NTA His Bind Resin affinity chromatography. His-OPN 13 fusion protein specifically inhibited adhesion and migration of VSMCs stimulated by osteopontin in dose-dependent manner. CONCLUSION: The OPN 13 peptide is successfully expressed in E.coli DH5α. The purified His-OPN 13 fusion protein could inhibit the adhesion and migration of VSMCs stimulated by osteopontin.     

Effects of apolipoprotein (a) on vascular smooth muscle cells proliferation and cell signal transduction pathway

AIM: To investigate the effect and the mechanism of apolipoprotein (a) ［apo (a)］ on proliferation of vascular smooth muscle cells (VSMCs). METHODS: All VSMCs used in experiments were serial subcultured from primary cells and were identified by immunohistochemistry staining of α-actin. Cell growth assay was observed as cell counting and MTT assay. Western blotting was also employed to detect the related mechanism. RESULTS: All cells used in experiments were confirmed as VSMCs. Although apo (a) enhanced VSMCs proliferation, this effect was attenuated by anti-integrin αⅤβ3, LM609. Use these reagents alone had no effect on VSMCs growth. The results of Western blotting demonstrated that focal adhesion kinase (FAK) was activated by apo (a) and the expression of total or phosphorylated transforming growth factor β1 (TGF-β1) was also decreased. However, these effects described above were all blocked by LM609. CONCLUSION: Apolipoprotein (a) enhances VSMCs proliferation and this effect is mediated by integrin αⅤβ3, which activates FAK and attenuates TGF-β1 and phospho-TGF-β1 expression.