凡纳滨对虾4个选育群体遗传多样性的SSR分析

采用9对微卫星引物分析了凡纳滨对虾4个选育群体(Molokai、OI、SIS和Kona Bay)的遗传多样性。结果表明:9对微卫星引物共检测到32个等位基因,每个位点等位基因数2～7个,平均3.555 6个,平均有效等位基因数2.472 7;各位点的观测杂合度(Ho)0.026 0～0.623 4,期望杂合度(He)0.263 1～0.785 5;各群体平均观测杂合度从小到大依次为SIS(0.383 3)相似文献   

军曹鱼全人工繁殖群体遗传特征的SSR分析

利用8个微卫星DNA位点分析南海海域5个军曹鱼全人工繁育群体(HN1、HN2、ZJ、FJ和LS)子代的遗传多样性特征和群体间遗传分化。结果显示,军曹鱼养殖群体与天然群体的遗传结构特征基本一致:1)平均有效等位基因数为3.910±0.440,观测杂合度为0.595±0.049,分子方差分析(AMOVA)表明,个体内分化占主导(46%),军曹鱼养殖群体整体遗传多样性较高;2)群体间基因流明显(N_m=2.5959,F_(st)=0.0878),整体分化程度较低。各养殖群体表现出不同于天然大群体的特征:1)绝大部分位点均明显偏离哈温平衡,杂合子缺失或过剩现象普遍存在;2)聚类和群体分配分析等表明HN2与另四个养殖群体(HN1、ZJ、FJ和LS)分化明显。     

马氏珠母贝4种壳色家系遗传差异的SSR分析

采用6对微卫星分子标记,分析了马氏珠母贝(Pinctada martensii)白、黑、黄、红4种壳色家系的遗传结构。结果表明:4种壳色家系在6个位点的平均等位基因数为4.17,有效等位基因数为1.297~1.819;平均观测杂合度为0.083~0.212,平均期望杂合度为0.207~0.341;Shannon多样性指数为0.366~0.616;平均多肽信息(PIC)含量为0.257~0.442;4个壳色家系哈-温平衡指数均发生了极显著的偏离,4个家系间的群体再分效应的固定指数FST值为0.113~0.793;与马氏珠母贝普通养殖群体及壳色选育系F5相比,4种壳色家系之间遗传距离变大,遗传多样性降低。     

罗非鱼4个选育群体遗传结构SSR分析

采用SSR分子标记分析了吉富品系罗非鱼的两个家系(GF1和GF2)以及奥利亚罗非鱼(Fo)和奥本尼罗罗非鱼(Fn)群体的遗传结构。结果显示,扩增后等位基因数为3～8个,随引物不同而异,14对引物共扩增60个等位基因,扩增片断大小在102～267 bp之间。微卫星分析表明,奥本尼罗罗非鱼(Fn)的平均观测杂合度(0.764 3)和平均期望杂合度(0.519 6)均最高,吉富尼罗罗非鱼(GF1)平均多态信息含量(0.419 5)最高;吉富尼罗罗非鱼(GF2)平均观测杂合度(0.614 2),奥利亚罗非鱼(Fo)的平均期望观测度(0.446 6)、平均多态信息含量(0.359 2)最低;因此,吉富鱼(GF1)的遗传多样性最高,奥利亚的遗传多样性最低。Hardy-Weinberg平衡遗传偏离指数(D)奥利亚(Fo)和尼罗(Fn)(0.463 4和0.478 9)明显高于吉富的两家系(0.234 1和0.250 0)。Fo与GF2间遗传距离(0.477 0)最大;而GF1和GF2间的遗传距离(0.302 7)最小,遗传相似系数(0.607 3)最大,可推断新一代吉富罗非鱼与本地选育群体有相对较远的亲缘关系,更具杂种优势。     

流沙湾卵鳎微卫星标记的遗传多样性分析

为分析湛江流沙湾海域优势渔种卵鳎的遗传多样性,应用微卫星标记技术,选用15对微卫星引物,以等位基因数、基因杂合度、多态信息含量、固定指数等遗传参数为指标,评估卵鳎群体内的遗传多态性。结果表明:共检测到90个等位基因,等位基因数从1~12不等,平均为6.0;有效等位基因数从1.0~8.4,平均为4.0,多态性位点比例为53%,显示其具有中等杂合子水平,其中8个多态位点的期望杂合度(He)为0.670~0.881,平均为0.800,观测杂合度(Ho)为0.353~1.000,平均为0.773,多态信息含量(PIC)值为0.616~0.870,平均为0.773,群体内固定指数F为-0.199~0.564,平均为0.046;流沙湾卵鳎群体具有高度遗传多样性。     

基于18S-ITS1-5.8S序列的吉富罗非鱼群体遗传多样性分析

运用PCR技术扩增吉富罗非鱼选育群体第20和21世代18S-ITS1-5.8S序列,分析二序列的遗传变异。结果表明,18S-ITS1-5.8S序列中,18S、内转录间隔区(ITS)1和5.8S序列分别为156、483~540、64 bp,主要变异位点位于ITS1中;基于ITS1序列的第20代吉富罗非鱼(17尾)群体内遗传距离为0.001,保守位点530个,变异位点10个,单倍型4个,单倍型多样性为0.331±0.143,核苷酸多样性为0.002,平均核苷酸差异为1.279;基于ITS1的第21代吉富罗非鱼(42尾)群体内遗传距离为0.015,6个个体存在严重碱基缺失现象,保守位点492个,变异位点49个,单倍型12个,单倍型多样性为0.638±0.083,核苷酸多样性为0.014,平均核苷酸差异为6.945。ITS序列在该两吉富罗非鱼群体中变异较小,基于ITS1序列分析的两代群体遗传多样性水平较低。     

胡子鲇3个养殖群体遗传结构的AFLP分析

应用AFLP方法,从64对引物中筛选出5对用于分析胡子鲇(Claris fuscus)厦门(XM)、江门(JM)、玉林(YL)等3个养殖群体共90个个体群体内、群体间遗传多样性。扩增共得到289条清晰谱带,引物多态性介于47.8%～63.8%,平均多态性比率为56.4%。胡子鲇群体总遗传杂合度(Ht)、遗传分化系数(Hs)、遗传分化度(Gst)和基因流(Nm)分别为0.161 1、0.153 6、0.051 1和8.501 8,表明胡子鲇多态性处于中等偏下水平,群体内部遗传分化系数较高,三个群体间无较大分化,群体间基因流动较好。其中,JM群体多态性比率、有效等位基因数Ne、Nei’s指数和Shannon指数都较XM群体和YL群体高,种内分化程度较好。对群体进行两两比较发现,XM群体和YL群体间分化程度最大,XM群体和JM群体次之,JM群体和YL群体最小。基于杂交优势,推断XM和YL群体繁殖所产生的后代将具有比亲代更为优良的性状。     

基于近缘EST-SSR引物的企鹅珍珠贝养殖和野生群体的遗传标记

利用近源物种马氏珠母贝的EST-SSR引物对企鹅珍珠贝DNA进行扩增,获得7对可扩增出目标条带的近缘分子标记,分析野生群体(YS)和养殖群体(YZ)企鹅珍珠贝的遗传多样性。结果显示,7个位点共获得26个等位基因;野生和养殖群体的等位基因数分别为3~5和3~4,平均期望杂合度(He)分别为0.623 5、0.496 8;平均观测杂合度(Ho)分别为0.431 8、0.302 8;HNUPM047位点的多态信息含量较低,为0.446 7,其他位点均在0.5以上,说明该遗传标记可提供比较丰富的遗传信息。表明养殖群体遗传多样性水平低于野生群体。     

黃鳍棘鲷放流苗种的遗传质量评估

【目的】评估黃鳍棘鲷(Acanthopagruslatus)增殖放流苗种的遗传质量。【方法】对黃鳍棘鲷放流苗种和自然群体的遗传多样性水平、遗传分化和近交程度等进行对比分析。【结果】放流苗种群体的等位基因数目NA(均值为9.917)大于自然群体NA(均值为8.083);而放流苗种群体的观测杂合度HO(均值为0.646)、期望杂合度HE(均值为0.694)和多态信息含量PIC(均值为0.654)均小于自然群体的HO(均值为0.651)、HE(均值为0.707)和PIC(均值为0.659)。此外,放流苗种群体和自然群体之间的存在一定的遗传分歧,遗传分化指数FST为0.011,但分歧程度不高,且近交程度系数均值F和FIS均大于自然群体的对应系数。【结论】黃鳍棘鲷放流苗种存在一定遗传质量缺陷,不能满足增殖放流要求,可能会自然群体产生负面影响。     

大口黑鲈肠道益生菌的分离与鉴定

【目的】分离鉴定大口黑鲈（Micropterus salmoides）肠道益生菌,评估分离菌用作饲料添加剂的益生潜力。【方法】采集大口黑鲈肠道样本,采用稀释涂布法分离细菌,经16S rRNA序列比对和生化试验鉴定种属,并进行抗逆性能、黏附能力、溶血活性和药敏特性等鉴定。【结果】从大口黑鲈肠道分离菌株84株,经测序、blast比对,筛选出3株同源性最高的菌株MS-1、MS-2、MS-3,分别鉴定为地衣芽孢杆菌（Bacillus licheniformis）、解淀粉芽孢杆菌（Bacillus amyloliquefaciens）和贝莱斯芽孢杆菌（Bacillus velezensis）。3株分离菌均为γ-溶血,不具有生物膜,疏水性50%～93.3%,自凝集性43.6%～69.2%;可在pH 2.0～12.0环境和5.0 g/L胆盐中存活;经高温处理后仍可生长;3株菌对大多数抗生素敏感。【结论】芽孢杆菌MS-1、MS-2和MS-3抗逆性高,黏附性能及饲料学安全性高,可作为大口黑鲈养殖中潜在益生菌。     

马氏珠母贝黄壳色选育系G_1遗传结构的ISSR分析

挑选11个黄壳色的马氏珠母贝养殖个体为亲本繁殖子代群体,并随机选取50个体为亲本,建立对照群体。采用ISSR分子标记对马氏珠母贝黄壳色群体和对照群体的遗传结构差异进行分析。结果表明,黄壳色和对照群体的遗传多样性分别为0.358 7和0.373 3,多态位点比例分别为86.89%和90.16%;群体间遗传分化系数为3.75%。可见,经过一代选择后,黄壳色群体遗传结构指标略低于对照群体,但两个群体未发生明显分化。     

大口黑鲈抗菌肽hepcidin cDNA序列和结构分析

以大口黑鲈为材料,提取肝脏总RNA,经RT-PCR扩增出hepcidin cDNA的开放阅读框(ORF)及3′端非编码区序列,应用5′RACE方法得到大口黑鲈hepcidin cDNA5′末端。将所获得的两个片段分别克隆到T载体后进行测序,并拼接成大口黑鲈hepcidin全长cDNA。序列分析表明:大口黑鲈hepcidin全长cDNA为564bp,含有一个258bp的ORF,编码86个氨基酸残基,由信号肽(24个残基)、前肽(42个残基)和成熟肽(20个残基)3部分组成hepcidin前体。在前肽部分具有前肽转化酶典型的RX(K/R)R基元,成熟肽部分含有8个保守的半胱氨酸残基,可形成四个链内二硫桥,使β-折叠结构保持稳定。大口黑鲈Hepcidin与其他鱼类的同源性在29.7%~90.5%间,尤其是信号肽区域,与鳜、尼罗罗非鱼、真鲷、花鲈、黑鯛、金眼狼鲈仅有2~3个氨基酸的差别。     

Mass Selection for Fast Growth in the Third Generation of the Orange Line of the Manila Clam Ruditapes philippinarum

The Manila clam Ruditapes philippinarum is natively distributed along the Pacific coast of Asia, where it is one of the most important bivalve species for local fisheries. This species has been intentionally introduced due to its high adaptability to various coastal environments and is currently widely cultivated in Europe and North America. We have initiated a selective breeding program to improve the growth rate of the Manila clam since 2007, and a full-sib family with orange shell color was selected. This family features high survival but slow growth. Thereafter, two generations of mass selection were conducted in this family by 10% upward selection for faster growth. In 2011, three types of lines were produced by selecting the breeder from the second-generation of selected lines. These lines are SS(subject to three generations of selection for growth in shell length), SC(selected for the second but not the third generation), and C(randomly sampled individuals from the national population). The genetic parameters of the shell length including hR2(realized heritability), SR(selection response), and GG(genetic gain) were analyzed. Results showed that the shell length of the SS line was significantly larger than those of the SC and C lines at all developmental stages(P 0.05). For the larval stage, the values of hR2, SR, and GG were 0.42, 0.73, and 6.66, respectively. For the juvenile stage, the values of hR2, SR, and GG were 0.40, 0.69, 21.76, respectively. For grow-out stage, the values of hR2, SR, and GG were 0.48, 0.83, and 18.22, respectively. The results of hR2 indicated the good potential of the SS line in selective breeding. The level of GG improvement was encouraging and consistent with the previous expectations. No inbreeding depressions in the shell length were observed in the SS and SC lines at all sampling days. All of the results indicate that maintaining selection pressure in successive generations may be effective in the selective breeding program of this family.     

大口黑鲈养殖研究

四年来，在研究了大口黑鲈生物学特性的基础上，吸收了国内外的养殖经验，结合实际确立了在水泥池和池塘中繁殖鱼苗、培育鱼种和池塘主养、配养食用鱼的养殖技术。     

Protective efficacy evaluation of immunogenic protein AHA_3793 of Aeromonas hydrophila as vaccine candidate for largemouth bass Micropterus salmoides

Journal of Oceanology and Limnology - Aeromonas hydrophila is a Gram-negative pathogen that can infect various fish, including largemouth bass (Micropterus salmoides), which have caused huge...     

大口黑鲈肌肉生长抑制素前肽真核表达载体的构建及其在肌肉组织中的表达

将大口黑鲈(Micropterus salmoide)肌肉生长抑制素(Myostatin,MSTN)前肽(MSTN-Pro)的cDNA定向克隆到真核表达载体pcDNA3.1(-)/mycHisB中,双酶切检测和测序鉴定证实,插入pcDNA3.1(-)/mycHisB载体中的片段为目的基因的核苷酸序列,MSTN基因前肽cDNA为正向插入,且重组质粒无错配或插入移位等突变。采用肌肉注射法将重组表达质粒注入大口黑鲈背部肌肉组织,在注射后第2天经RT-PCR检测到MSTN前肽基因mRNA的表达,第6天经免疫组化学检测到MSTN前肽蛋白的表达,第8天蛋白表达强度增强,对照组始终未检测到MSTN前肽基因的表达。     

Analysis of genetic diversity and differentiation of seven stocks of Litopenaeus vannamei using microsatellite markers

Seven microsatellite markers were used to evaluate the genetic diversity and differentiation of seven stocks of Litopenaeus vannamei, which were introduced from Central and South America to China. All seven microsatellite loci were polymorphic, with polymorphism information content(PIC) values ranging from 0.593 to 0.952. Totally 92 alleles were identified, and the number of alleles(Na) and effective alleles(Ne) varied between 4 and 21 and 2.7 and 14.6, respectively. Observed heterozygosity(Ho) values were lower than the expected heterozygosity(He) values(0.526–0.754), which indicated that the seven stocks possessed a rich genetic diversity. Thirty-seven tests were detected for reasonable significant deviation from Hardy-Weinberg equilibrium. Fis values were positive at five loci, suggesting that there was a relatively high degree of inbreeding within stocks. Pairwise Fst values ranged from 0.0225 to 0.151, and most of the stock pairs were moderately differentiated. Genetic distance and cluster analysis using UPGMA revealed a close genetic relationship of L. vannamei between Pop2 and Pop3. AMOVA indicated that the genetic variation among stocks(11.3%) was much lower than that within stocks(88.7%). Although the seven stocks had a certain degree of genetic differentiation and a rich genetic diversity, there is an increasing risk of decreased performance due to inbreeding in subsequent generations.