缢蛏糖原磷酸化酶基因多态性与糖原含量的关联分析

为探究缢蛏糖原磷酸化酶基因(Sc-GPH)与糖原含量的关系,本研究克隆获得Sc-GPH cDNA全长,检测其在不同组织、不同月份的表达模式,并在Sc-GPH基因编码区筛选与糖原含量关联的SNP位点.结果表明,Sc-GPH cDNA全长为3963 bp,开放阅读框为2541 bp,编码846个氨基酸;氨基酸序列多重比对和系统发育树显示,缢蛏与欧洲大扇贝、虾夷扇贝、长牡蛎等贝类亲缘关系较近,而与哺乳类、甲壳类、昆虫类亲缘关系较远.实时荧光定量PCR结果显示,Sc-GPH在8个组织中均有表达,其中在外套膜和足中表达量最高(p<0.01),推测与其糖原存储能力有关;不同月份的缢蛏外套膜和足中,Sc-GPH在8月表达量最高,而此时糖原含量较低,说明Sc-GPH的表达可能受缢蛏生殖周期的影响.以缢蛏"甬乐1号"群体为实验材料,在Sc-GPH基因编码区筛选到4个与糖原含量相关SNP位点,其中c.930T>C位点在台州野生群体中得到进一步验证,该位点为高糖原缢蛏分子标记辅助选育提供了候选标记.     

香港牡蛎糖原合酶与糖原磷酸化酶基因多态性研究及糖原含量性状相关分子标记挖掘

香港牡蛎是我国南方沿海地区的主要经济贝类,糖原含量是衡量其品质高低的重要指标之一。本研究在转录组测序基础上,以三墩群体(SD,60只个体)和大风江群体(DFJ,90只个体)为研究对象,采用ELISA方法检测每个香港牡蛎糖原含量,并利用片段长度差异等位基因特异性PCR (Fragment length discrepant allele specific PCR,FLDAS-PCR)技术开展糖原合酶和糖原磷酸化酶基因多态性研究,开发单核苷酸多态性(Single nucleotide polymorphism,SNP)标记。结果共开发25个SNPs,其中4个SNPs (Ch_GS680,Ch_GS1248,Ch_GP992和Ch_GP1962)在实验群体中分型成功,SPSS关联分析显示SNP位点Ch_GS680与糖原含量显著关联(P<0.05),暗示其可作为香港牡蛎糖原含量性状的潜在分子标记,研究结果为高糖原含量香港牡蛎新品种的分子标记辅助选育奠定了基础。     

糖原含量与糖原磷酸化酶基因(GPH)和己糖激酶基因(HK)的表达在福建牡蛎生殖周期中的相关性分析

Glycogen, a polymer of glucose, is an important means of storing energy. It is degraded by glycogen phosphorylase (GPH) and hexokinase (HK), glycogen phosphorylase, and hexokinase cDNAs (Ca-GPH andCa-H...     

九孔鲍MSTN基因cSNP多态性及与生长性状关联分析

九孔鲍(Haliotis diversicolor supertexta)是中国南方具有较高经济价值的养殖贝类。肌肉生长抑制素(Myostatin, MSTN)是转化生长因子β超家族(TGF-β)中参与动物肌肉生长的重要调控因子, 因此MSTN基因是动物遗传改良的重要候选基因。为探究九孔鲍MSTN基因的多态性及其生长相关性, 实验采用PCR产物直接测序方法对九孔鲍MSTN基因进行单核苷酸多态性(SNP)筛选, 对MSTN基因SNP与体质量、壳长、壳宽进行关联性分析, 运用实时荧光定量PCR (qRT-PCR)技术检测九孔鲍MSTN基因不同基因型的表达水平。结果显示: 在九孔鲍MSTN基因中共筛选出9个SNP位点。SNP位点与生长性状关联分析发现, 九孔鲍MSTN基因编码区SNP位点g909C>T发生C/T同义突变与九孔鲍体质量、壳长、壳宽显著相关。TT基因型九孔鲍的体质量显著高于CC基因型和TC基因型个体; TT基因型的壳长、壳宽显著高于CC基因型(p<0.05)。通过qRT-PCR检测显示, 在九孔鲍MSTN基因SNP位点g909C>T的3种基因型中, TC和CC基因型的基因表达量显著高于TT基因型(p<0.05)。研究结果表明MSTN基因SNP位点g909C>T可作为九孔鲍标记辅助选择育种重要候选标记。     

缢蛏(Sinonovacula constricta)EST-SSR标记与生长性状的相关性分析

利用16个微卫星标记对缢蛏F1家系的遗传多样性和标记-性状相关性进行了研究。结果显示,16个SSR位点共检测到39个等位基因,平均等位基因数(Na)和平均有效等位基因数(Ne)分别为2.44和2.09;多态信息含量(PIC)为0.40;平均观测杂合度(Ho)与期望杂合度(He)分别为0.58和0.63,部分位点基因型分布严重偏离孟德尔定律,暗示其可能与适应性基因相连锁,其中YC-8位点附近可能存在杂合不利存活基因。位点YC-96和YC-123与体重呈显著相关(P0.05),YC-96的BC基因型和YC-123的BB基因型均为体重性状的优势基因型,其对应的壳长、壳宽、壳高和体重4个生长性状表型值都最大,可以作为选育快长性状的候选分子标记;YC-1位点与壳宽呈显著相关(P0.05),其中BB型是壳宽的优势基因型;YC-93位点与壳宽、壳高呈极显著相关(P0.01),其中AB型是壳高的优势基因型,BB和BC型是壳宽的优势基因型。对YC-1、YC-93、YC-96和YC-123进行以体重性状为参照的不同基因型组合比较,找到了最优组合(BB/BB(BC)/BC/BB),与4个位点单独分析对应的最优基因型基本完全一致,符合加性作用模型。筛选出的与生长性状相关的标记可为标记辅助选择育种(MAS)提供参考依据。     

三门湾养殖缢蛏中重金属元素的含量及其食用安全性

采用多种分析方法,对浙江省三门湾养殖的缢蛏Sinonovacula constricta Lamarck重金属元素含量进行了分析,并与象山港养殖的大黄鱼Pseudosciaena crocea、海带Laminaria japonicaAre-sch中相应的重金属元素含量进行了比较,探讨了其食用安全性,结果表明:(1)缢蛏中含重金属Cu、Pb、Zn、Cd、Cr、As、Hg和Se元素的量分别为:(16.4±0.8),(1.29±0.13),(159±8),(0.38±0.02),(0.36±0.05),(12.5±0.6),(0.057±0.004),(3.16±0.15)mg/kg,其中Cu和Zn元素的含量较高,而海带和大黄鱼中Hg元素的含量较高,但均符合国家《农产品安全质量无公害水产品安全要求》中限定标准的要求,可见缢蛏、大黄鱼和海带都能从水体中吸收、富集重金属元素,但对重金属元素吸收和富集的能力是有一定差异的。(2)海产品中所含的重金属元素主要是天然存在或是由人为污染所导致的,适量的Cu和Zn元素对动物和人体的生存和生长是有益的,但Pb、Cd和Hg等有毒的重金属元素应控制在国家限制标准之内。将干样换算成湿样可见,缢蛏中Cd元素的含量已接近国家标准的限值,因此应引起养殖部门的密切关注和高度重视,适时采取措施,避免超标。     

缢蛏耐高盐性能及其与生长性状的相关性分析

缢蛏(Sinonovacula constricta)是我国重要的水产经济贝类,开展耐高盐新品种选育对平衡不同地区缢蛏良种养殖意义重大。本研究对不同家系缢蛏幼贝进行高盐胁迫,运用统计学方法和相关性分析探究了缢蛏耐高盐性能及其与生长性状的相关性。结果显示,缢蛏家系间壳长、壳高、壳宽和体质量表型值差异较大,体质量变异系数达15.53%。不同家系对高盐耐受性差异较大,家系间死亡率范围在0～100%。胁迫实验共持续9 d,各家系缢蛏首粒死亡时间在第1～9 d之间,而且死亡率较高的家系内缢蛏死亡时间离散程度较大,说明不同个体间高盐耐受性差异也较大。缢蛏开始出现大量死亡的时间主要在胁迫后第8 d,第9 d部分家系死亡率高于80%,说明缢蛏对高盐耐受存在一个阈值,超过该阈值会开始或大量死亡。皮尔逊和斯皮尔曼相关性分析显示,缢蛏存活时间和生长性状为正相关,其中存活时间与缢蛏壳长、体质量相关性均达到显著水平(P<0.05)。缢蛏存活率与体质量、生长速度均显著正相关(P<0.05)。整体而言,壳长较长和体质量较重的缢蛏个体耐高盐性能较强,在高盐环境中具有生长优势。本研究揭示了缢蛏的高盐耐受性及...     

缢蛏消化道组织学与组织化学的研究

运用组织学和组织化学染色方法对缢蛏消化道的组织结构和功能进行了研究。结果表明 ,缢蛏的消化道由唇瓣、口、食道、胃、晶杆囊、肠、直肠和肛门组成。除唇瓣外的消化管壁由内向外可分为 4层 :粘膜层、粘膜下层、肌层和外膜。粘膜层包括粘膜上皮和粘膜肌层。口腔上皮为假复层纤毛柱状上皮 ,其它部位的粘膜上皮都为单层纤毛柱状上皮。胃壁的局部和肠壁缺乏肌层。除口的腹面、直肠、肛门等处的外膜为浆膜外 ,其余皆为纤维膜。肠上皮、直肠上皮和唇瓣上皮有较强的碱性磷酸酶活性 ,胃上皮和肠上皮有较强的酸性磷酸酶活性。直肠的粘膜及粘膜下层中含有丰富的钙和铁     

饥饿和恢复投喂对金鳟体组分和糖原含量的影响

研究在(16±2)℃的条件下饥饿和恢复投喂不同时间后金鳟(Golden Rainbow Trout,Oncorhynchus mykiss)体组分和糖原含量的变化.试验鱼分为4个组,每组设3个平行,分别饥饿7 d,恢复投喂44 d(S7组);饥饿14 d,恢复投喂37 d(S14组);饥饿21 d,恢复投喂30 d(S21组),对照组(S0组)正常喂食.研究发现:饥饿使金鳟的水分和灰分含量升高,粗脂肪和糖原含量显著下降,粗蛋白含量无显著变化.饥饿7 d后,肝糖原和肌糖原含量与试验前和同期对照组相比下降极显著(P<0.01).饥饿至14 d,水分含量上升,与试验前和同期对照组有极显著差异(P<0.01),灰分含量与试验前和同期对照组相比上升显著(P<0.05),粗脂肪含量与试验前和对照组下降极显著(P<0.01).饥饿至21 d,水分含量与试验前和同期对照组相比上升显著(P<0.05),肝糖原和肌糖原含量均有不同程度的回升,但仍与试验前和同期对照组存在极显著差异(P<0.01).恢复投喂后,除S21组肌糖原含量仍极显著低于对照组(P<0.01)外,其他各组的各指标经统计学分析均恢复到与对照组无显著差异水平(P<0.05).结果表明:金鳟饥饿过程中动用储存物质的顺序是首先动用糖原然后动用脂肪,而蛋白质含量变化不明显.金鳟在贮存能源物质时优先积累肝糖原.     

Reproductive investment of the American razor clam Ensis americanus in the Dutch Wadden Sea

The American razor clam Ensis americanus is a marine bivalve indigenous from the American coast that was introduced in Europe in the late 1970s and has managed to build up strong populations along the European coast. In the present paper, we studied energy investment into soma and gonads by following the seasonality in growth and reproduction of this species in a subtidal area of the western Dutch Wadden Sea. Body and somatic mass indices showed a seasonal pattern with maximal values in August and minimal in April. Gonadal mass index showed the opposite trend with highest values in April. Spawning started in April/May and a second, but weaker, spawning event apparently occurred in August/September. Gonadosomatic ratio values were very low. Just before the spawning period, only 2.5% of the body mass of E. americanus consisted of gonads.     

宁波长街缢蛏遗传结构的RAPD分析

利用RAPE)技术对宁波长街自然群体和养殖群体缢蛏【Sinonovacula constricta(Lamarck)】的遗传背景进行了比较研究。21个引物共扩增出121条清晰可辨的RAPE)标记,大小在250--2500bp之间,分别统计上述位点,得到两群体各遗传参数。结果表明,养殖环境对其影响较小,两者问遗传距离仅为0．0497,近交系数(Fst)为0．0735,有效繁殖个体数(Ncm)为3．1514;两者平均遗传杂合度比较却发现养殖群体略大于野生群体。与其他的水产动物研究结果不同。作者推测上述结果与养殖缢蛏苗种来自天然采苗、无累代养殖和养殖区饵料较自然海区丰富等因素有关。     

Gametogenic development and spawning of the razor clam,Zenatia acinaces in northeastern New Zealand

The potential exists for New Zealand to exploit already established markets for razor clams through development of fisheries or aquaculture industries for the New Zealand razor clam, Zenatia acinaces. However, fishery or aquaculture development for Z. acinaces requires an understanding of the reproductive cycle including the timing of gametogenic development and spawning. The reproductive cycle of Z. acinaces was studied over an 11‐month period from May 2000 to March 2001 at Kennedy Bay, Coromandel Peninsula, using qualitative standard histological analysis and quantitative measures of oocytes. Histological analysis indicated that Z. acinaces is dioecious and gametogenic development was synchronous between the sexes. Gametogenesis began in June with gametes maturing quickly and by August/September (late winter/ early spring) most razor clams were ripe. Spawning began as early as September (spring) although spawning mainly occurred during October. By December (summer), nearly all clams were completely spent. From January 2001 most clams could not be sexed as all residual gametes were resorbed. Razor clams remained in this stage during March 2001. Spawning began when the water temperature was around its lowest, c. 15°C. Monthly mean number of eggs/follicle was sensitive to changes in reproductive development, closely following patterns observed in the qualitative stagings. Patterns of monthly mean oocyte diameters did not adequately describe the spawning events observed in qualitative analyses. Sex ratios were equal over the size range (69–99 mm shell length) of clams that could be sexed. The data presented in this study provide valuable information on the timing of spawning events for Z. acinaces, necessary for developing sustainable management strategies and selecting broodstock for aquaculture.     

The effect of PCBs on glycogen reserves in the eastern oyster Crassostrea virginica.

Recent declines in Chesapeake Bay oyster populations have been attributed to disease, and reduced water quality from pollution. The stress associated with pollutant exposure may reduce energy available for growth and reproduction. Polychlorinated biphenyls (PCBs) are lipophilic contaminants that may potentially affect mobilization of lipid reserves, increasing reliance on glycogen stores, which could otherwise be utilized to supply energy for gametogenesis. Thus, PCBs may indirectly affect glycogen stores in oysters in a deleterious manner. To test for this effect, reproductively inactive oysters were exposed to PCBs by feeding individuals 0.7 g of algal paste containing 0, 0.35, or 3.5 micrograms PCBs daily for 8 weeks. Additionally, a group of oysters was exposed to PCBs (0, 0.35, and 3.5 micrograms) plus 0.3 g of non-toxic artificial sediment to examine interactive effects of sediment particles and PCBs. Adductor muscle, mantle, and gonadal tissues were analyzed for glycogen content. Results suggest that glycogen content is reduced in the adductor muscle with increasing PCB exposure, but there are no effects of PCBs in the mantle and gonadal tissues.     

廉州湾红树蚬的繁殖生物学研究

2011年8月至2012年7月,采集广西北海廉州湾红树蚬(Polymesoda erosa)的周年样品进行了性腺发育、生殖周期和胚胎发育研究。结果表明,红树蚬雌雄异体,性别可通过成熟生殖腺颜色区分;1年1个生殖周期,性腺发育可分为增殖期、生长期、成熟期、排放期和休止期5个阶段,性腺发育两性同步;繁殖期从6月延续到11月,其中7月和10月为繁殖高峰期;肥满度指数最高为5.19%(5月份),最低为1.98%(2月份);繁殖方式为非孵育型,沉性卵,外被一层厚的胶质卵膜,水温26℃时,D形幼虫孵出时间约为27h。