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1.
建立黑斑原鮡(Glyptosternum maculatum)AFLP反应体系,对DNA提取、双酶切、连接、预扩增、选择性扩增和银染效果进行了探索。结果表明,40μLTaqI酶切体系中65℃先酶切2h,EcoRI酶切和连接同时进行,EcoRI酶和T4DNA连接酶的用量均以2U为宜,温度与时间分别为37℃和3h,连接产物稀释10倍作为预扩增的模板。利用这种优化的反应体系,5对选择性扩增引物组合表现出较高的稳定性、清晰度和多态性,它们分别是:E-AAC/T-AAT、E-AAC/T-AAG、E-AAG/T-ACA、E-AAG/T-AAT和E-AAT/T-AGA。5对引物组合对96个样本扩增,共得到332条清晰可辨、可重复的条带,平均每对引物组合扩增出66.4条带,其中51条带是多态的。将AFLP技术用于黑斑原鮡的研究,通过优化体系的建立,为其种质资源的研究奠定基础。  相似文献   

2.
以珠江长臀(Cranoglanis bouderius)为试验材料,建立了长臀扩增酶切片段长度多态性(AFLP)分析体系,对其分析过程中的DNA提取、双酶切、单酶切、连接、预扩增、选择性扩增、银染等步骤进行了检测,并对反应条件进行了优化。用PstI/TaqI、EcoRI/MseI等2组双酶切组合构建长臀的AFLP指纹图谱,条带清晰可辨,扩增信号强,得到清晰的长臀AFLP指纹图谱。  相似文献   

3.
大菱鲆AFLP分析体系的建立   总被引:5,自引:0,他引:5  
本研究构建了大菱鲆扩增片段长度多态性(AFLP)分析体系,对DNA提取、双酶切反应、连接反应、预扩增反应、选择性扩增反应和银染等步骤进行了分析。结果表明,用EcoRI/MseI双酶切、核心序列加3个选择性碱基的选择性扩增引物、1∶6(w∶w)的EcoRI端与MseI端选择性扩增引物比和20μl选择性扩增体积,可得到十分稳定的结果。所得产物经电泳和银染后可获得条带清晰、背景干扰小的图像。该体系的构建为AFLP技术在大菱鲆相关研究中的应用奠定了基础。  相似文献   

4.
程飞  叶卫  叶富良  刘波 《南方水产》2007,3(4):46-51
以珠江长臀(鮠)(Cranoglanis bouderius)为试验材料,建立了长臀(鮠)扩增酶切片段长度多态性(AFLP)分析体系,对其分析过程中的DNA提取、双酶切、单酶切、连接、预扩增、选择性扩增、银染等步骤进行了检测,并对反应条件进行了优化.用PstⅠ/TaqⅠ、EcoRⅠ/MseⅠ等2组双酶切组合构建长臀鲍的AFLP指纹图谱,条带清晰可辨,扩增信号强,得到清晰的长臀(鮠)AFLP指纹图谱.  相似文献   

5.
本研究构建了西伯利亚鲟(Acipenser baeri)扩增片段长度多态性(AFLP)分析体系,分析了DNA提取、双酶切反应、连接反应、预扩增反应、选择性扩增反应和银染等步骤.用7对引物对24尾西伯利亚鲟进行了AFLP分析,每对引物扩增的位点数在24~39之间,共扩增出了234个位点,多态性位点为116个,多态性比例为...  相似文献   

6.
甘肃金鳟RAPD反应体系构建及优化   总被引:1,自引:0,他引:1  
以甘肃金鳟尾鳍为试验材料,提取基因组DNA。对影响甘肃金鳟RAPD扩增的重要参数进行了优化试验,包括模板DNA浓度、dNTP浓度、引物浓度、Taq酶、变性时间、退火温度等,建立了甘肃金鳟RAPD反应的最适体系。即在25μl反应体系中:模板DNA用量为4.0 ng/μl;Mg2 浓度为2.5mmol/L;dNTP浓度为0.5 mmol/L;引物浓度为0.5μmol/L;Taq酶用量为1 U;扩增程序:94℃预变性4 min,94℃变性45 s,36℃退火50 s,72℃延伸1 min,共35个循环,最后延伸10 min。  相似文献   

7.
本研究构建了西伯利亚鲟(Acipenser baeri)扩增片段长度多态性(AFLP)分析体系,分析了DNA提取、双酶切反应、连接反应、预扩增反应、选择性扩增反应和银染等步骤。用7对引物对24尾西伯利亚鲟进行了AFLP分析,每对引物扩增的位点数在24~39之间,共扩增出了234个位点,多态性位点为116个,多态性比例为48.83%,平均Nei氏基因多样性指数H为0.4297,平均Shannon氏指数I为0.1233,个体间平均遗传距离为0.1872,表明西伯利亚鲟养殖群体的遗传多样性程度比较低,同时还构建了西伯利亚鲟的聚类图。  相似文献   

8.
以魁蚶为研究材料,建立了魁蚶的AFLP分子标记反应体系。对其分析过程中的酶切链接、预扩增、选择性扩增等试验条件进行了检测与筛选,得到了清晰的AFLP指纹图谱,为探讨魁蚶种群之间的遗传关系和遗传图谱的构建等方面提供了新的技术手段。  相似文献   

9.
以赣江野生青鱼(Mylopharyngodon piceus)基因组DNA为试验材料,研究Mg2+浓度、dNTP浓度、引物浓度和模板浓度4个参数对ISSR-PCR反应的影响,建立一套适合青鱼ISSR-PCR反应的最佳体系。结果表明,在25μL的反应体系中,Mg2+、dNTP、引物和模板4个参数的最适宜浓度分别为:3.0mmol/L、0.25mmol/L、0.4umol/L和30ng。PCR扩增程序为:94℃预变性5min,94℃30s,52℃45s,72℃2min,共38个循环,最后72℃继续延伸10min。  相似文献   

10.
用超分支滚环扩增法检出传染性脾肾坏死病毒。超分支滚环扩增法包括锁式探针的连接及超分支滚环扩增法扩增两部分,通过研究锁式探针的最佳连接体系和连接条件,并进行超分支滚环扩增法反应条件的优化,得出的本研究最适反应条件为:锁式探针在T4 DNA连接酶作用下37℃连接20 min,接下来的超分支滚环扩增法在Bst DNA聚合酶大片段作用下61℃反应40 min,即能够实现靶序列的有效检出。灵敏度试验表明,超分支滚环扩增法所能检测到的最低模板量接近1 copy。在4种病毒中进行的特异性试验表明,本方法能够保证对传染性脾肾坏死病的特异性检测。  相似文献   

11.
为获知野生、池塘及工厂化养殖马口鱼肌肉营养成分的差异,实验分析了野生、池塘及工厂化养殖马口鱼的形态学指标、肌肉营养成分、氨基酸含量、脂肪酸含量、理化特性及重金属含量。结果显示,野生马口鱼肌肉中苯丙氨酸、天冬氨酸、必需氨基酸总量、非必需氨基酸总量及滴水损失显著高于池塘马口鱼;野生马口鱼脏体比、肌肉粗蛋白、支链氨基酸/芳香族氨基酸、C16:0及熟肉率显著低于工厂化马口鱼,但水分、甲硫氨酸、含硫氨基酸、C18:3n6、饱和脂肪酸、铅、镉和铜的含量显著高于工厂化马口鱼;野生马口鱼肥满度、肌肉中缬氨酸、亮氨酸、苏氨酸、赖氨酸、异亮氨酸、谷氨酸、酪氨酸、精氨酸、氨基酸总量、半必需氨基酸总量、鲜味氨基酸、苦味氨基酸、呈味氨基酸、C20:0、C20:2、C22:2、C22:1n9及C20:5n3的含量与工厂化马口鱼无显著性差异。研究表明,相比野生和池塘养殖,工厂化养殖可以提高马口鱼肌肉粗蛋白、氨基酸总量、C20:5n3、熟肉率及滴水损失,减少饱和脂肪酸、镉 、铅及铜,是一种较优的马口鱼养殖模式。  相似文献   

12.
2003年5月~2008年4月,对李仙江的鱼类资源进行了调查。共采集鱼类标本7195号。结合历史记录,李仙江共记录鱼类64种(亚种),分属于6目14科43属,其中6种为外来种。李仙江鱼类区系中种类最多的是鲤形目的鲤科,共24属31种;其次为鲤形目爬鳅科和鲇形目鮡科,各为4属7种。近年考察数据显示,李仙江土著鱼类资源呈下降趋势。为保护李仙江土著鱼类资源,应建立鱼类自然保护区和鱼类人工增殖放流站,加强李仙江流域环境保护、生物多样性监测和渔业管理。  相似文献   

13.
鲟鱼病原性疾病研究进展   总被引:1,自引:0,他引:1  
鲟是地球上最古老的物种之一。我国自1991年后开始鲟类的商业化养殖,主要品种有引进的匙吻鲟、杂交鲟、小体鲟、俄罗斯鲟和西伯利亚鲟,黑龙江流域出产的施氏鲟、达乌尔鳇,以及施氏鲟和达乌尔鳇的杂交种等。随着养殖业的发展,鲟疾病不断增多,严重制约了养殖产业的健康发展。目前发现的鲟致病性病原有病毒、细菌、真菌和寄生虫等。概述了目前国内外鲟病原性疾病的研究进展,包括感染种群、致死率、临床症状、病原分离鉴定和药物控制等方面。目前对鲟病害的研究,在发病机制、免疫防治以及结合环境和营养因素的综合防控技术等方面还十分缺乏,应系统开展鲟疾病的流行病学、病原学、诊断技术以及防控技术研究。  相似文献   

14.
Extractive components were determined for the mantle muscle and liver of three species of Loliginidae squid, Sepioteuthis lessoniana, Loligo bleekeri, and L. edulis, and one species of Ommastrephidae, Todarodes pacificus, as a control. Total free amino acid levels and the major free amino acids in muscle, taurine, proline, glycine, alanine, and arginine, were significantly higher in Loliginidae squids than in T. pacificus. The main nucleotide was adenosine 5′-monophosphate, which did not differ significantly between species. Malate was the organic acid found in muscle in highest concentrations. The muscle of these species contained a large amount of trimethylamine oxide. A large amount of glycine betaine was also detected in the muscle, but showed no large species difference. From these results, the muscle of Loliginidae species is considered to have a much sweeter taste than that of T. pacificus. Compared with muscle, squid liver was characteristic with high contents of taurine, glutamate, bitter amino acids, succinate, propionate, trimethylamine, and glycine betaine, and with low contents of sweet amino acids, arginine, nucleotides, malate, and trimethylamine oxide. These results suggest that squid liver is characterized by a complicated taste containing umami, bitterness, sourness, fishy flavor, and less sweetness.  相似文献   

15.
Approximately 5000 young of the year (0+) cyprinids comprising roach, chub, dace, minnow, bleak, bream, barbel and gudgeon were examined histologically for the presence of myxozoan infections. Thirteen myxozoans were identified to species, the majority being Myxobolus spp. In addition, two species of Myxidium and of Sphaerospora were recorded. All organs were examined, with the majority of infections being found in the gills, musculature and kidney. However, isolated spores were occasionally found in other tissues. Whilst roach contained the highest number of myxozoan species, it was chub that showed the greatest host response to sporogonic forms. Data are provided on spore morphology, pathogenic responses and tissue and host specificity of the myxozoans recorded.  相似文献   

16.
宋凯  郭远明 《河北渔业》2012,(5):5-8,59
对我国几种传统海洋药用动物中重金属元素的含量进行了测定,并对同产地的陆生药用动物蛤蚧进行比较研究,其中Hg和As利用原子荧光光度计测定,Pb、Cd、Cu、Fe、Mn、Zn利用原子吸收光度计测定,样品采用微波消解仪消解。结果表明:海钱、海蛇、海马、海星的镉含量较高,海燕、海马、海星的铅含量较高,除个别样品微量元素未检出外,样品中Fe、Mn、Zn的含量相对较高。最后对金属元素含量较高的原因及对所含微量元素的食用安全性和营养学评价进行了探讨。  相似文献   

17.
  1. The status of marine biodiversity in the Eastern Central Atlantic (ECA), especially of coastal and pelagic fishes, is of concern owing to a number of threats including overharvesting, habitat loss, pollution, and climate change combined with inadequate policy responses, legislation, and enforcement.
  2. This study provides the first comprehensive documentation of the presence, status, and level of extinction risk, based on IUCN Red List assessment methodology, for more than 1800 marine species, including all taxonomically described marine vertebrates (marine mammals, sea turtles, seabirds, fishes); complete clades of selected marine invertebrates (sea cucumbers, cone snails, cephalopods, lobsters, reef‐building corals); and marine plants (mangroves, seagrasses).
  3. Approximately 8% of all marine species assessed in the ECA are in threatened categories, while 4% are listed as Near Threatened, 73% are Least Concern, and 15% are Data Deficient. Fisheries and overharvesting are the biggest threats to living marine resources in the ECA, with 87% of threatened species across all taxonomic groups affected by both large‐ and small‐scale targeted fisheries, excessive capture as by‐catch, or unsustainable harvest.
  4. The results of this study will transform the current state of knowledge and increase capacity for regional stakeholders to identify and enact marine conservation and research priorities, as a number of species are identified as having high conservation and/or research priorities in the region.
  5. Through the process of marine species data collection and risk assessments conducted over the past 5 years, several key conservation actions and research needs are identified to enable more effective conservation of marine biodiversity in the ECA, including increased governance, multilateral collaboration, taxonomic training, and improved reporting of fisheries catch and effort.
  相似文献   

18.
This study evaluated the suitability of commercially produced fermented soybean meal (FSBM) known as PepSoyGen?, in a plant‐based diet for Florida pompano, Trachinotus carolinus fingerlings. An 8‐week growth trial was conducted to evaluate the effect of four isonitrogenous and isolipidic diets containing 0, 206, 309 and 410 g/kg FSBM, replacing approximately 0%, 50%, 75% and 100% SBM (designated as Basal, FSBM 50, FSBM 75 and FSBM 100, respectively) on growth performance, body composition, serum biochemistry and morphological condition of liver and distal intestine of Florida pompano. There were no significant differences in final mean weight, percentage weight gain, thermal unit growth coefficient and feed conversion ratio in all treatments. For serum biochemistry analysis, there were no significant differences in total protein, albumin, glucose, cholesterol, bile acids, plasma alkaline phosphatase, alanine aminotransferase and aspartate aminotransferase activities in all treatments. Cellular infiltration, presence of goblet cells and the width of lamina propria in the distal intestine were partly improved in fish fed FSBM 75 and 100 compared to the fish fed FSBM 50 and basal diet. Glycogen granulation, inflammation and nuclear change condition in the liver of pompano were better as the inclusion of fermented product increased. Results of this study indicate that FSBM can be utilized as an alternative protein source and microbial fermentation process could improve the functional properties of SBM.  相似文献   

19.
中国介螅水母属二新种(丝螅水母目,介螅水母科)   总被引:1,自引:1,他引:0  
记述了台湾海峡及其邻近海区介螅水母科Hydractiniidae L.Agassiz,1862介螅水母属Hydractinia van Beneden,1841二新种,即螺旋介螅水母,新种Hydractinia spiralis sp. nov.和反曲介螅水母,新种Hydractinia recurvatus sp.nov.,兹将二新种主要鉴别特征描述于下:螺旋介螅水母,新种:伞无顶突;垂管长圆柱形,约为内伞腔深度2/3;口腕简单,末端具成束刺胞;生殖腺很大,呈椭圆形,几乎覆盖整个垂管间辐位,无水母芽;缘触手6条,4条主辐位,另2条仅在相对间辐位,基球延长锥状,同样大小,无眼点,触手长,近端2/3粗壮,远端1/3变细,具螺旋状刺胞。反曲介螅水母,新种:伞无顶突;垂管短小,呈方形;胃柄显著,约为垂管长度1/2;口有4个主辐位口唇,延长成向上反曲的短口腕,末端具成丛刺胞;4个卵圆形生殖腺,附在二个反曲口腕之间的垂管间辐位,无水母芽;缘触手16条,触手基球膨大,呈锥状,同样大小,整条触手具环状刺胞,末端略膨大,呈椭圆形。  相似文献   

20.
Piscimetrics deals with software implementation of experimental design, second-generation artificial intelligence tools, viz. Neural Nets (NNs), genetic algorithms, Fuzzy Logic, Expert Systems, Wavelets and Image analysis in the field of fisheries. A brief sketch of NNs is followed by a review of their applications in forecasting, classification, distribution and fisheries management since 1978. Forecasting in fisheries covers distribution of eggs, recruitment, fish growth/age, biomass and fish catch. Other major areas are identification, abundance and food products, environmental factors and collapse of fishery industry. The data structures are given in tensorial notation. The need for the paradigm shift from classical to multi-level hybrid NNs is emphasized.  相似文献   

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