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1.
利用Roche 454 GS FLX平台测序技术进行了钝吻黄盖鲽微卫星引物筛选,并采用聚类分析的方法对得到的5641个微卫星位点进行类比分析,得到247种多态性位点,其中完美型占52.22%,非完美型占20.24%,复合型占27.54%,(AC)n、(AG)n两碱基重复类型的比例是44%,重复次数在10次以上的占总数的87.5%。随机选取11个位点的微卫星引物,采用5个野生个体,利用荧光标记和毛细管电泳进行多样性评价,4个位点偏离Hardy-Weinberg平衡,不同位点得到的等位基因范围为3-8,平均等位基因数为5.0。平均观望杂合度(Ho)、平均期望杂合度(He)及平均多态性信息含量(PIC)分别为0.588、0.788和0.670。结果表明,454 GS FLX提供了一种直观、高效开发微卫星的方法。  相似文献   

2.
利用Roche 454 GS FLX平台测序技术进行了钝吻黄盖鲽微卫星引物筛选,并采用聚类分析的方法对得到的5641个微卫星位点进行类比分析,得到247种多态性位点,其中完美型占52.22%,非完美型占20.24%,复合型占27.54%,(AC)n、(AG)n两碱基重复类型的比例是44%,重复次数在10次以上的占总数的87.5%。随机选取11个位点的微卫星引物,采用5个野生个体,利用荧光标记和毛细管电泳进行多样性评价,4个位点偏离Hardy-Weinberg平衡,不同位点得到的等位基因范围为3?8,平均等位基因数为5.0。平均观望杂合度(Ho)、平均期望杂合度(He)及平均多态性信息含量(PIC)分别为0.588、0.788和0.670。结果表明,454 GS FLX提供了一种直观、高效开发微卫星的方法。  相似文献   

3.
采用磁珠富集法筛选适合漠斑牙鲆遗传多样性分析的微卫星分子标记。筛选共获得43条序列,其中完美型26个,占60.5%;非完美型14个,占32.6%;复合型3个,占6.9%。选取其中14对特异性好且扩增效率高的微卫星引物,对采自美国北卡罗来纳州沿海的漠斑牙鲆野生群体和养殖群体进行遗传多样性及遗传结构比较分析。研究结果表明,12对引物的扩增产物具有多态性,其中7个位点为高度多态(PIC>0.5)。两个群体中共检测到90个等位基因。12个多态性微卫星位点在两个群体中的平均观察杂合度(Ho)和期望杂合度(He)分别为0.36和0.57。9个位点在整个群体中呈现出不同程度的偏离遗传平衡(P<0.05),且偏离平衡的位点均表现为杂合子缺失(Fis>0)。野生群体和养殖群体间的遗传距离为0.1115,群体间的遗传分化微弱(Fst=0.0438)。  相似文献   

4.
通过对长体圆鲹(Decapterus macrosoma)基因组进行RAD-Seq高通量测序,共获得58 180条微卫星序列,选取112条二、三核苷酸重复的微卫星序列设计引物,经筛选后,共获得27个具有多态性的微卫星标记。利用一个长体圆鲹群体对通过筛选的微卫星标记的种群遗传学特征进行评价。结果显示,27对引物扩增的序列中18个位点为二核苷酸重复,重复次数为9~14次,9个位点为三核苷酸重复,重复次数为6~10次,等位基因数(Na)为5~17 (平均10.6),表观杂合度(Ho)为0.342 9~0.857 1 (平均0.631 7),期望杂合度(He)为0.538 3~0.911 8(平均0.796 8),多肽信息含量(PIC)为0.497~0.886 (平均0.780 9),除1个位点外,其他位点PIC值均大于0.500,表明开发的微卫星位点具有较高的多态性。"哈迪-温伯格"平衡(HWE)检测结果显示,19个标记等位基因频率符合HWE。连锁不平衡检测表明各位点间无连锁不平衡现象。该研究开发的27个微卫星标记可为长体圆鲹种群遗传学研究提供基础。  相似文献   

5.
仿刺参微卫星标记的筛选及群体遗传结构分析   总被引：1，自引：0，他引：1       下载免费PDF全文
采用FIASCO(Fast Isolation by AFLP Sequences Containing Repeats)方法构建了仿刺参基因组富集CA微卫星序列的基因组短片段文库,筛选得到118条微卫星DNA序列。根据重复单元的排列特点,完美型共83个,占70.4%;非完美型共30个;占25.4%;复合型标记5个,占4.2%。选取其中20条微卫星序列设计引物进行多态性检测,并利用这20对引物对采自中国、韩国(西海岸及东海岸)、俄罗斯和日本沿海的野生仿刺参以及美国沿海的具疣拟刺参进行遗传多样性水平和遗传结构分析。结果表明,所设计的20对引物的扩增产物均具有多态性,其中16个位点为高度多态(PIC>0.5)。遗传多样性分析结果显示,20个微卫星座位的平均观察杂合度(Ho)和平均期望杂合度(He)分别为0.39和0.69,共检测到231个等位基因(102个有效等位基因),在每个座位上获得3～20个等位基因,平均等位基因数为11.6个,20个位点在6个群体中不同程度偏离遗传平衡(P<0.05),所有群体在整体上均表现为杂合子缺失(Fis>0);5个仿刺参群体间的遗传相似系数在0.71以上,相似性较高,而仿刺参群体与具疣拟刺参的相似性则较低。聚类分析结果表明,中国群体与韩国西海岸群体聚类成一支,而俄罗斯群体、韩国东海岸群体和日本群体聚类成另外一支,聚类的先后与它们在地理分布上的海域位置有一定的相关性。  相似文献   

6.
贾舒雯  刘萍  李健  李吉涛  高保全  陈萍  潘鲁青 《水产学报》2012,36(12):1819-1825
利用12对微卫星标记分析了莱州湾(LZ)、海州湾(HZ)、象山(XS)脊尾白虾野生群体的遗传多样性.12个位点在3个群体中均表现出高度的多态性.12个微卫星位点共得到115个等位基因,各个位点的等位基因数介于6～14,平均每个位点上的等位基因数为9.583 3个;各个位点的平均期望杂合度(He)、平均观测杂合度(Ho)、平均多态信息含量(PIC)分别为0.8278、0.517 5、0.705 1,表明3个脊尾白虾野生群体均有良好的多态性.经F-统计分析,各个群体间的遗传分化指数均值为0.093 0(0.05＜Fst＜0.15),呈中等水平分化.基于Nei's遗传距离的UPGMA聚类分析显示莱州湾群体与象山群体距离最近,聚为一类,海州群体单独聚为一类.  相似文献   

7.
草鱼三、四核苷酸重复微卫星标记的分离与特征分析   总被引：4，自引：1，他引：3  
采用磁珠富集法、通过质粒检测法结合同位素杂交检测获得草鱼(Ctenopharyngodon idellu)三、四核苷酸阳性克隆1 378个,测序705个,共得到846个微卫星座位,其中完美型632个（占74.7％）,非完美型114个(占13.48％),混合型100个(占11.82％).根据微卫星侧翼序列设计并合成100对微卫星引物,检测结果显示35对(35％)引物在邘江野生群体中表现出多态性.选择其中2对多态性稳定的引物进行遗传多样分析,结果显示,20个位点共检测到212个等位基因,平均观察杂合度(Ho)为0.681 1,平均期望杂合度（He）为0.797 5,平均多态信息含量为0.777 7.结果表明,所筛选的微卫星标记能够用于草鱼群体遗传学研究.本研究旨在为草鱼的种群遗传结构分析、遗传图谱构建等研究奠定基础.  相似文献   

8.
富集文库－菌落原位杂交法筛选栉孔扇贝的微卫星标记   总被引：1，自引：1，他引：0  
应用微卫星富集文库─菌落原位杂交法,筛选得到了40个栉孔扇贝的微卫星标记.用固定了(AG)15和(AC)15探针的尼龙膜(Hybond N )捕捉含有微卫星DNA的片段,经洗脱、PCR扩增和TA克隆,构建栉孔扇贝的微卫星富集文库.利用ECL试剂盒(Amersham公司)标记的(AG)15和(AC)15探针进行菌落原位杂交筛选微卫星富集文库,阳性克隆经测序获得微卫星DNA.富集文库中1200个重组克隆经过菌落原位杂交后,532个(44.3%)为阳性克隆.任意挑选100个克隆测序,结果显示所有的克隆都至少含有一个微卫星位点.利用软件设计了65对特异性PCR引物,40对能扩增出清晰的带谱;利用48个栉孔扇贝个体评价微卫星位点,分析表明37个位点具有多态性.不同的位点获得的等位基因数目为2～14个不等,37个多态性位点共获得258个等位基因,平均每个位点获得7.0个等位基因.观测杂合度(Ho)、期望杂合度(He)及多态性信息含量值(PIC)的范围分别为0.1000～1.0000、0.1197～0.9831和0.1172～0.9782.结果表明,富集文库─菌落原位杂交法适合大规模筛选目标物种的微卫星标记.  相似文献   

9.
采用Illumina高通量测序技术对银鲴(Xenocypris argentea)肝脏组织进行转录组测序分析,筛选微卫星标记并进行多态性检验。共获得7 272个微卫星位点。随机选取48个4碱基以上重复类型的SSR位点进行引物设计和PCR验证,有47对可以扩增出清晰稳定的条带。在上述47个位点中随机选择26对在洞庭湖银鲴群体中进行多态性分析,结果表明,具有多态性的微卫星引物为21对。不同多态位点得到的等位基因数范围为3~15,平均基因数为7.29±3.94,观测杂合度(Ho)、期望杂合度(He)、多态信息含量(PIC)以及香浓-威尔指数(H)平均值分别为0.532 7±0.211 4、0.613 6±0.196 4、0.564 6±0.198 0和1.302 0±0.577 9。21个微卫星位点中,有6个显著偏离哈代-温伯格平衡(Hardy-Weinberg equilibrium,HWE)。  相似文献   

10.
根据前人FIASCO方法构建的三疣梭子蟹微卫星富集文库,对含微卫星的DNA序列设计了71对引物并对其进行了多态性筛选,筛选出21对多态的微卫星引物,对三疣梭子蟹1个野生群体的30个个体进行遗传多样性检测,同时对引物进行评价。21个位点共获得了188个等位基因,平均每个位点扩增得到8.9个等位基因。不同引物获得的等位基因数差异较大,从3～13个不等,其中Pot8、Pot37、Pot48、Pot53、Pot54、Pot66六个位点分别获得了11、12、12、11、13、11个等位基因,而Pot46仅获得了3个等位基因。等位基因的大小分布在131～312bp,基本符合引物设计时理论产物长度。21个微卫星位点的期望杂合度的范围为0.659～0.889,PIC值均高于0.5,表明它们都有很高的杂合度,均可用于三疣梭子蟹种群遗传结构分析,为三疣梭子蟹品种选育、种系评估提供更多的微卫星DNA信息。  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.
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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.
The endemic, anadromous cyprinidChalcalburnus tarichi is the only fish species known to occur in alkaline Lake Van (Eastern Anatolia, Turkey). EightC. tarichi were maintained individually in Lake Van water (17 – 19°C; pH 9.8; 153 mEq·I–1 total alkalinity; 22 total salinity) and tank water samples analyzed for 24 h in 2 to 4 h intervals. At zero time, < 1µM ammonia was present and urea was undetectable in the tank water; at 24 h, total ammonia and urea made up 114±32 and 35±25µM, respectively. Over the experimental period, ammonia-N and urea-N excretion averaged 1041±494 and 607±169moles·kg–1 fish·h–1, respectively. The extent of urea excretion was highly variable between specimens. Uric acid excretion was not detectable.Urea was present at high concentrations in all tissues and plasma (25 – 35moles·g–1·ml–1) of freshly caughtC. tarichi; total ammonia content of the tissues was by a factor of 1.9 (liver) to 3.0 (brain) lower. High arginase activity (2.4±0.2 U·min–1·g–1) was detected in the liver ofC. tarichi but ornithine carbamoylphosphate transferase, a key enzyme of the ornithine-urea-cycle, was absent. Ureagenesis is likely through degradation of arginine and/or uricolysis. High glutamine synthetase activity (11±0.6 U·min–1·g–1) and low ammonia content in brain suggest that, like other teleosts,C. tarichi has an efficient ammonia detoxification in the brain, but in no other tissue.Nitrogenous waste excretion at alkaline pH is discussed. The ability ofC. tarichi to excrete high levels of ammonia at extremely alkaline pH is unique among teleosts studied so far. The mechanism of ammonia excretion under Lake Van conditions remains to be elucidated.  相似文献   

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