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1.
用RAPD技术对5种经济海胆基因组DNA多态性的研究   总被引:10,自引:0,他引:10  
利用随机扩增多态DNA技术对紫海胆(Anthocidaris crassispina)、海刺猬(Glyptocidaris crenularis)、马粪海胆(Hemicentrotus pulcherrimus)、光棘球海胆(Strongylocentrotus nudus)和中间球海胆(Strongylocentrotus intermedius)等我国最主要的5种经济类海胆基因组DNA的多态性进行初步研究一经20个随机引物扩增,共得到377条扩增片段.片段长度为200~1600bp。根据片段的共享度计算出平均遗传距离并采用CPTGMA和NJ两种聚类分析方法进行处理得到系统树。从聚类图得知长海胆科与疣海胆科有较近的亲缘关系,球海胆科的光棘球海胆与中间球海胆有较近的亲缘关系,其次是马粪海胆。本研究还讨沦了RAPD技术在海胆的遗传学研究中的广阔应用前景以及从分子生物学方法上为海胆传统分类方法提供科学依据。  相似文献   

2.
5种经济海胆线粒体16SrRNA基因片段的序列分析   总被引:1,自引:0,他引:1  
对光棘球海胆、中间球海胆、马粪海胆,海刺猬和紫海胆线粒体DNA 16SrRNA基因片段进行了PCR扩增和测序,得到395 bp的可信片段。通过ClustalX 1.83和Mega 3.0软件对所得线粒体16SrRNA片段序列进行比较,共检测到118个碱基变异,其中简约信息位点为33个。应用Mega 3.0计算各种间的相对遗传距离,以糙海参为外类群,得到NJ系统树和MP系统树,系统树各分支的置信度由“Bootstrap”1000次循环检验。结果表明,马粪海胆与光棘球海胆的亲缘关系较近,其次为中间球海胆,而紫海胆、海刺猬与这3种海胆的关系相对较远。  相似文献   

3.
不同种海胆体腔细胞类型及体液中的酶活力   总被引:1,自引:0,他引:1       下载免费PDF全文
以中国主要经济海胆马粪海胆(Hemicentrotus Pulcherrimus)、虾夷马粪海胆(Strongylocentrotus intermedius)、光棘球海胆(Strongylocentrotus nudus)、紫海胆(Anthocidaris crassispina)以及杂交种海胆-马粪海胆(♀)×虾夷马粪海胆(♂)、虾夷马粪海胆(♀)×光棘球海胆(♂)、马粪海胆(♀)×紫海胆(♂)和紫海胆(♀)×光棘球海胆(♂)为对象,进行海胆的血细胞及体液中酶活力的研究。结果表明,海胆血细胞可分为4类,分别为色素细胞、纤毛游走细胞、变形吞噬细胞和无色球形细胞,比例分别为4.56%~10.96%、9.69%~12.18%、76.40%~85.36%和0.35%~1.03%;海胆血细胞体外有凝聚和吞噬现象。不同种海胆体液中酸性磷酸酶(ACP)、碱性磷酸酶(ALP)、超氧化物歧化酶(SOD)和溶菌酶(LSZ)的酶活力测定结果显示,紫海胆血清中各种酶的活性都较高,其中碱性磷酸酶、超氧化物歧化酶活性在6种海胆中都是最高的,分别为2.871 U/mg和182.149 U/mL。马粪海胆(♀)×虾夷马粪海胆(♂)的溶菌酶活性最高,为0.085 U/mL;虾夷马粪海胆(♀)×光棘球海胆(♂)的酸性磷酸酶活性最高,为1.070 U/mg,但差异均不显著(P>0.05)。[中国水产科学,2006,13(1):33-38]  相似文献   

4.
为获得Y-box蛋白在中间球海胆的生物学功能,实验克隆了中间球海胆Y-box基因的全长cDNA,并进行了定量表达模式分析.Y-box基因全长cDNA为2 425 bp,该序列具有一个81 bp的5’非编码区,1 348 bp 3’非编码区,ORF为996 bp,cDNA编码331个氨基酸的前体蛋白,预测蛋白分子质量为37.4 ku,理论等电点是8.55,属于亲水性蛋白.中间球海胆Y-box前体蛋白的氨基酸序列包含3个结构域:氨基酸N末端,亲水结构域C末端和冷休克结构域(cold shock domain CSD),其中CSD区为Y-box蛋白家族中保守结构域.海胆Y-box蛋白质二级结构中无规则卷曲占88.22%,延伸链占9.97%,α-螺旋1.98%,无β-折叠.氨基酸序列与其他物种氨基酸序列的同源性为22.37%~41.26%,在保守结构域CSD区同源性达到75%以上.实时定量PCR检测,Y-box基因在中间球海胆体腔液、管足、围口膜、雄性性腺、雌性性腺、肌肉和肠7个不同组织中均有表达,但差异不显著.温度胁迫下,中间球海胆Y-box基因的表达均表现为下调,并且随着温度的升高,Y-box基因的表达量呈现逐渐下降的趋势.  相似文献   

5.
王波 《齐鲁渔业》2002,19(9):3-3
球海胆属(Sh。楷叫优。ntrotu抓9海胆世界上共有十几种,光棘球海胆(S.n。;d u s)和中间球海胆(S.fll。。。。巾l。)是我国北方主要的经济品种,其营养价值和经济价值高,国际市场需求量大,开展海胆人工育苗和增养殖是海胆产业可持续发展的重要举措。而掌握其胚胎和幼虫生长发育的区别,不仪对苗种生产有帮助,而且对海胆资源的保护利用十分有利。 1卵及胚胎 光棘球海胆的卵径为106 pITI左右,黄色;中间球海胆90 pill左右,橙色,其卵径比光棘球诲胆小一点。受精膜举起后,光棘球海胆的卵径为  相似文献   

6.
中间球海胆乳酸脱氢酶基因克隆及其对海水酸化的响应   总被引:1,自引:0,他引:1  
为明确中间球海胆乳酸脱氢酶(LDH)基因序列及表达特征,研究其对海水酸化胁迫的响应,实验利用cDNA末端快速扩增(RACE)技术获得中间球海胆LDH基因(命名为SiLDH)的全长cDNA,并且比较了海水酸化胁迫下,中间球海胆不同组织SiLDH基因表达及总LDH活性的变化情况。结果显示:①SiLDH基因的cDNA全长为1 499 bp,包含133 bp的5′非编码区,349 bp的3′非编码区和1 017 bp编码338个氨基酸的开放阅读框(ORF)。SiLDH编码蛋白的相对分子量为36.40 ku,理论等电点为6.55,属于无信号肽的非跨膜、温和疏水蛋白。②生物信息学分析显示,SiLDH蛋白序列与紫球海胆LDH X4蛋白序列一致性最高(90.88%)。③实时荧光定量PCR(qRT-PCR)发现,SiLDH基因在检测的4种组织中均有表达,相对表达量从高到低为性腺管足肠体腔液;总LDH活性检测显示,中间球海胆总LDH活性从高到低为性腺管足体腔液肠;④海水酸化处理60 d后发现,与自然海水组(pH 8.06±0.01)相比,SiLDH基因在中间球海胆性腺、管足和肠道中呈现总体降低趋势,在体腔液中则呈现随海水pH降低先降低后显著升高的趋势;总LDH活性在性腺、管足和体腔液中呈现随海水pH降低而降低的趋势,在肠道中则呈现随海水pH降低先降低后显著升高的趋势。研究表明,中间球海胆面对海水酸化时可能通过调节SiLDH基因的表达和LDH活性来缓解海水酸化带来的负面影响。  相似文献   

7.
四种海胆杂交的可行性及子代的早期发育   总被引:18,自引:0,他引:18  
常亚青 《水产学报》2000,24(3):211-216
报道了中国北部沿海马粪海胆、海刺猬、光棘球海胆三种主要海胆与引自日本的中间球海胆四种海胆之间的不同组合的杂交试验及其子代浮游幼体及幼海胆的早期生长发育.结果表明,采用生殖调控可使不同海胆达到同步繁殖,在8~24℃下各种海胆杂交组合的受精率与亲本亲缘关系有关,同时受到双亲繁殖适宜温度的影响,受精率介于0~69.6%之间,均低于自交组.以马粪海胆和海刺猬为亲本的正反交、以马粪海胆和中间球海胆为亲本的正反交以及马粪海胆()×光棘球海胆()各组杂交胚胎均可发育至四腕幼虫,但畸形率较高,浮游时间比自交组延长3~10d,在其中的3个杂交组得到了幼海胆,经过14个月的室内培育,杂交组成活率低于自交组,壳直径达2.59~2.88cin.  相似文献   

8.
采用PCR扩增测序法获得了棘头梅童鱼Collichtys iucidus南通、启东、芦潮港、温州、瑞安、福州、番禺7个地理群体53尾鱼的mtDNA控制区全序列.棘头梅童鱼控制区序列长度为778~782 bp,序列长度变异主要是由位点的插入或缺失造成,识别了终止序列区TAS和保守序列区CSB1、CSB2、CSB3序列.共检测到多态位点66个,占全部序列的8.44%,检测到单倍型33种,平均单倍型多样性(Hd)为0.934 7±0.017 5,平均核苷酸多样性(π)为0.017 5±0.002 3.棘头梅童鱼群体内的遗传距离(K 2-P)为0.001 7~0.022 8,群体间的遗传距离为0.002 3~0.043 3,群体间的分化指数FsT为-0.058 4~0.845 8,基因流为0.05~19.62.AMOVA分析结果显示,群体间的变异为74.21%,群体内的变异为25.79%(P<0.05),表明群体间发生了显著的遗传分化.以NJ法构建的亲缘关系树显示,棘头梅童鱼7个群体分为两个大支,一支以北方类群(南通、启东、芦潮港、温州群体)个体为主,另一支以南方类群(福州和番禹群体)个体为主,而位于南、北方类群中间位置的瑞安群体则与两大类群均有交叉.  相似文献   

9.
以草鱼(Ctenopharyngodon idellus)为实验对象,利用RACE技术克隆了草鱼成纤维细胞成长因子8(Fibro-blast Growth Factor8,FGF8)cDNA序列。实验结果显示,草鱼FGF8 cDNA序列长度为887 bp,其中5′-非翻译区长100 bp,3′-非翻译区长157 bp,开放阅读框(ORF)633 bp,编码210个氨基酸的前体蛋白,包含22个氨基酸的信号肽和188个氨基酸的成熟肽。利用已知的FGF8序列绘制系统发育树,发现草鱼与两种鲤科鱼(班马鱼、墨西哥脂鲤)聚为一簇,显示亲缘关系很近,而与其它脊椎动物如两栖类、鸟类、哺乳类的亲缘关系则相对较远,这与形态分类学的结果一致。此外,还通过生物信息学分析,揭示了草鱼FGF8的分子结构特征。  相似文献   

10.
三峡库区3种银鱼线粒体DNA细胞色素b基因序列多态性分析   总被引:3,自引:0,他引:3  
对三峡库区大银鱼(Protosalanx chinensis)、太湖新银鱼(Neosalanx taihuensis)和短吻间银鱼(Hem isalanxbrachyrostralis)共78尾个体的Cyt b基因全序列(1141bp)进行了分析。结果显示,序列有16个变异位点,总变异率为1.4%,其中13个为简约位点。大银鱼的32尾样本有6个单倍型,太湖新银鱼32尾样本有5个单倍型,短吻间银鱼14尾样本中有3个单倍型。大银鱼、太湖新银鱼和短吻间银鱼的单倍型多样性指数(h)分别为0.804±0.032,0.671±0.061,0.385±0.149;核苷酸多样性指数(π)分别为0.00146±0.00009,0.00228±0.00024,0.00067±0.00026。3种银鱼群体的遗传距离在0.12461~0.23796之间,净遗传距离在0.12274~0.24418之间。基于Cyt b基因序列,利用K imura-2模型构建的NJ树表明:大银鱼与太湖新银鱼种群的亲缘关系最近,两者与短吻间银鱼种群的亲缘关系较远。研究结果表明,三峡库区3种银鱼的遗传多样性低。  相似文献   

11.
The environmental processes associated with variability in the catch rates of bigeye tuna in the Atlantic Ocean are largely unexplored. This study used generalized additive models (GAMs) fitted to Taiwanese longline fishery data from 1990 to 2009 and investigated the association between environmental variables and catch rates to identify the processes influencing bigeye tuna distribution in the Atlantic Ocean. The present findings reveal that the year (temporal factor), latitude and longitude (spatial factors), and major regular longline target species of albacore catches are significant for the standardization of bigeye tuna catch rates in the Atlantic Ocean. The standardized catch rates and distribution of bigeye tuna were found to be related to environmental and climatic variation. The model selection processes showed that the selected GAMs explained 70% of the cumulative deviance in the entire Atlantic Ocean. Regarding environmental factors, the depth of the 20 degree isotherm (D20) substantially contributed to the explained deviance; other important factors were sea surface temperature (SST) and sea surface height deviation (SSHD). The potential fishing grounds were observed with SSTs of 22–28°C, a D20 shallower than 150 m and negative SSHDs in the Atlantic Ocean. The higher predicted catch rates were increased in the positive northern tropical Atlantic and negative North Atlantic Oscillation events with a higher SST and shallow D20, suggesting that climatic oscillations affect the population abundance and distribution of bigeye tuna.  相似文献   

12.
In this experiment, a feeding trial was performed to determine the effects of fructooligosaccharide (FOS) on growth performance, digestive enzyme activity and immune response of Japanese sea bass, Lateolabrax japonicus juveniles (initial weight 38.3 ± 0.5 g), and the fish were examined following feeding with six levels of FOS (0, 0.5, 1, 2, 4 and 6 g/kg) for 28 days. Significant enhancement of weight gain (WG) and specific growth rate (SGR) was found in fish fed 1 g/kg FOS incorporated diets (p < .05), while the feed conversion ratio (FCR) in the 1, 2 g/kg FOS groups reduced significantly compared with the control (p < .05). Besides, the crude lipid in the 4, 6 g/kg FOS groups increased significantly compared with the control (p < .05). On the other hand, the erepsin and lipase activities significantly elevated in intestine of fish fed 2 g/kg FOS (p < .05) and the lysozyme activity in serum of fish fed 2 g/kg FOS were significantly higher than that in the control (p < .05). Moreover, the alkaline phosphatase activities in serum of fish fed 0.5, 1, 2 g/kg FOS were significantly higher than in control (p < .05). Regression analysis showed that the relationships between dietary FOS levels and either SGR, FCR, erepsin or lysozyme activities were best expressed by regression equations, and the optimal inclusion levels are 1.37, 1.80, 3.06, 3.11, 1.93 and 1.80 g/kg for SGR, FCR, erepsin, lipase, lysozyme and total superoxide dismutase activities, respectively. Overall, this study revealed that FOS incorporated diets could beneficial for L. japonicus culture in terms of increasing the growth, digestion and immune activities. Under the present experimental condition, the optimal supplementary level of FOS in the diet of L. japonicus is 1–3 g/kg.  相似文献   

13.
Previously, we reported 10 PEGE types of 85 tilapia Streptococcus agalactiae(GBS), which shifted from Streptococcus iniae in China, by using PEGE method. Presently, larger and more representative tilapia GBS were isolated, for the ?rst time in China, to characterize their serotypes and genetic diversities more precisely than had done before. 168 GBS strains were distributed in ?ve provinces of China, in which Guangdong, Guangxi and Hainan were the major ones, holding36.9%(62/168), 37.5%(63/168) and 19.6%(33/168), respectively. Serotypes, Ia, Ib and III, were observed in these strains and the most predominant one was Ia(95.2%), which mainly distributed in Guangdong, Guangxi and Hainan. Ia initially occurred in 2009, it shoot up to 32.1% in 2010,but decreased to 16.1% in 2011 before went up to 45.2% in 2012. Ib sporadically occurred during2007–2011, III onlyoccurred in 2012. 14 different PFGE types, including 4 new types(N, O,P and Q), were observed, in which B, D, F and G were the predominant types, holding 83.9%(141/168) of the total GBS strains. Ia corresponded to 11 PFGE types(A–H, N–P), in which type D predominated(51%). Ib represented 3 genotypes(I, J and Q) and III harbored only 2genotypes(N and F). Type N and Fsynchronously presented in Ia and III. In summary, the genetic diversity of tilapia GBS varied by serotypes and changed with geographical locations and years.Although Iastillpredominated, new rareserotypeIII alreadyoccurred in China.  相似文献   

14.
15.
Growth hormone (GH) and reproduction: a review   总被引:5,自引:0,他引:5  
Interaction between growth and reproduction occurs in many vertebrates and is particularly obvious at certain stages of the life cycle in fish. Endocrine interactions between the gonadotropic axis and the somatotropic axis are described, the potential role of GH being emphasised. A comparative analysis of these phenomena in mammals, amphibians and fish, suggests a specific role of GH in the physiology of puberty, gametogenesis and fertility. It also shows the original contribution made by studies on the fish model in this field of investigations.  相似文献   

16.
Two gonadotropins (GtH; Qa and Qb) were purified by gel filtration and ion exchange chromatography from the pituitaries of Indian walking catfish (Clarias batrachus). The presence of GtH during purification was assessed by in vitro oocyte maturation and in vivo steroidogenic activity, and their identities were determined by elution profiles, molecular weight, biological activities and yield. The molecular weights of Qa and Qb were 37 and 42 kDa, respectively, and composed of distinct subunits (Qa: 20 and 14 kDa and Qb: 26 and 18 kDa). Polyclonal antibodies raised against Qa immunostained Qa, Qb and pituitary GtH cells. A competitive Qa‐ELISA was developed whose sensitivity was 6.25 ng mL?1 (1.25 ng well?1) with intra‐ (3.5%) and inter‐ (12.4%) assay coefficients of variation. Displacement curves parallel to the standard were obtained with plasma and pituitary extracts of catfish, Qb and carp GtHII. The assay was validated by measuring the plasma Qa levels after LHRH treatment and in relation to ovarian growth in the female catfish during different reproductive phases. Based on the results, Qa and Qb corresponded to fish LH and FSH respectively. The findings will increase the knowledge of the mechanisms controlling fish reproduction and identification of sensitive phases in fish in captivity for hormonal manipulation.  相似文献   

17.
Tetrahymena is a protozoan parasite, which infects guppy, Poecilia reticulata Peters, and causes substantial economical losses in commercial farms worldwide. Studies of guppy infected by Tetrahymena require standardized infection protocols. The LD50 for Tetrahymena infection of guppies by intraperitoneal (IP) injection was calibrated, and the level obtained was 946 parasites per fish. Guppy infection with Tetrahymena by immersion, imitating the natural route of infection via the integument, was studied under normal or stress conditions. Exposure to cold and netting (CNI) and to cold only (CI) followed by immersion exposure to 10 000 Tetrahymena per mL resulted in 22.5% and 19.2% mortality, respectively, as compared to 14.2% and 10% in groups that were netted only (NI) or non‐stressed (I). Histopathology revealed that immersion infection resulted in a systemic infection. Lysozyme levels, measured 3 weeks after infection, were significantly higher in the CNI group (288 μg per mg protein) compared with CI‐, NI‐ and I‐treated groups (94.5, 64 and 62.3 μg mg?1, respectively). There was no evident parasite immobilization activity in body homogenates, suggesting no development of acquired immunity. Re‐infection by IP injection revealed no increase in protection in any of the treatment groups, mortality range of 56.3–75%, higher than in the non‐exposed control (40.6% mortality).  相似文献   

18.
Plasma estradiol-17 (E2), testosterone (T), 17,20-dihydroxy-4-pregnen-3-one (DHP) and 17,20,21-tri-hydroxy-4-pregnen-3-one (20-S) levels were measured by radioimmunoassay (RIA) in white perch (Morone americana) and white bass (M. chrysops) that were induced to undergo final oocyte maturation (FOM) with human chorionic gonadotropin (hCG). Plasma DHP levels increased in females of both species in association with oocyte germinal vesicle migration (GVM) and germinal vesicle breakdown (GVBD) and decreased thereafter. Plasma 20-S levels also increased with oocyte GVM in white bass, but were several-fold lower than DHP levels. Circulating E2 and T levels were greatest during GVM and GVBD in both species and decreased to low levels during oocyte hydration and ovulation. Follicles from white perch and white bass which received a priming injection of hCG in vivo, produced both DHP and 20-S in vitro after exposure to hCG and their oocytes underwent GVBD. Ovarian incubates from unprimed fish of either species produced only E2 and T and their oocytes did not complete GVBD. Oocytes from unprimed bass, but not perch, matured when follicles were exposed to hCG in vitro. Both trilostane and cycloheximide blocked in vitro production of DHP and 20-S and oocyte GVBD by white perch follices. DHP and 20-S were equipotent inducers of FOM in the GVBD bioassay. None of several other structurally-related steroids tested were effective within a physiological range of concentrations. These results indicate a role for DHP and 20-S in the control of FOM in white perch and white bass.  相似文献   

19.
Changes in heart rate, ventilatory activity and oxygen consumption were determined in trout (Salmo gairdneri) and brown bullhead catfish (Ictalurus nebulosus) during exposure to a steadily increasing concentration of waterborne cyanide selected to produce death in 8–9 hours for each species. The lethal cyanide concentration for the bullheads was an order of magnitude higher than for trout. Trout developed an immediate and gradually increasing bradycardia throughout the exposure period. Cyanide produced tachycardia in the bullhead followed by a gradual onset of bradycardia as the concentration of cyanide was raised. Pericardial injection of atropine (a muscarinic cholinergic antagonist) indicated that bradycardia in the trout was due initially to increased vagal tone but later due to the direct effect of cyanide on the heart. Hyperventilation in the trout persisted throughout the exposure period, although the rate and amplitude fluctuated and was variable between individual fish. During the last hour of exposure (highest cyanide concentration), ventilation was characterized by rapid, shallow breaths followed by a sudden respiratory arrest. The bullheads exhibited hyperventilation during the first 3 hours of exposure followed by a gradual, linear drop in ventilation rate and amplitude until death occurred. Cardiac and ventilatory responses in both species were attributed to stimulation of central and peripheral chemoreceptors by cyanide. Evidence is presented which suggests the initial response in the bullheads was due, at least in part, to gustatory stimulation by the cyanide. Oxygen consumption of the trout remained above pre-exposure levels for the majority of the test period. Oxygen consumption in the bullhead paralleled the changes in heart and ventilatory rates. Whole-body lactate levels of fingerlings of both species during cyanide exposure were measured to estimate the extent of anaerobiosis. Whole-body lactate levels were much greater in the bullheads than the trout, indicating a higher capacity for anaerobiosis, possibly due to a greater fuel supply. Overall, the trout responded to cyanide in a manner similar to that produced by environmental hypoxia whereas the bullheads experienced a gustatory stimulus which masked the hypoxia-like response.  相似文献   

20.
This study brings an integrated analysis about the relationship between water deterioration and its physiological consequences in live fish transport. The analysis was focused on the transport water and its deterioration, and physiological challenges imposed on the fish. Usual commercial handling procedures employed to mitigate fish stress during transport were discussed. Future topics of research for the establishment of safer fish transport protocols were proposed. Transport was classified into short (≤8 h) or long transport (>8 h). The main issue in short transports should be the prevention of water pH reduction, while in long transports it is the increase in ammonia. Plasma cortisol is the most employed marker for stress and is acutely elevated upon short episodes of transport, but remains elevated even in long‐transport events. Plasma glucose is perhaps a better marker for handling stress. Plasma lactate, pH, osmolality CO2 and ions should be more often evaluated. Plasma Na+ and Cl are very useful markers of acidosis, due to their respective exchange for H+ and , for acid–base regulation. The establishment of species‐specific transport protocols should be preceded by such combined analyses of water and physiological parameters.  相似文献   

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