基于SLAF-seq技术的黑棘鲷微卫星标记开发及其在鲷科鱼类中的通用性研究     

吴仁协  肖瑶  牛素芳  翟云  张浩冉  陈伟勇  黎晓 《海洋与湖沼》2019,50(2):365-377

黑棘鲷(Acanthopagrusschlegelii)是广泛分布于西北太平洋的暖温性中下层经济鱼类,也是我国沿海重要的海洋捕捞鱼类和增养殖对象。然而,目前有关黑棘鲷的微卫星标记研究报道较少,难以对其种质资源状况作出精确评估。本研究采用SLAF-seq技术测序共获得22489个二至六碱基重复的黑棘鲷微卫星序列,短重复序列(二、三碱基)占总微卫星序列的90.8%,长重复序列(四至六碱基)占有9.2%。经过157对随机合成引物的多态性筛选,开发出49个高多态性的黑棘鲷微卫星标记,其中短重复序列位点有25个,长重复序列位点有24个。每个位点的等位基因数(Na)为2—20(均值为8.3),观测杂合度(Ho)和期望杂合度(He)分别为0.097—0.938和0.122—0.922(均值分别为0.663和0.701),多态信息含量(PIC)为0.118—0.897(均值为0.655)。经Bonferroni校正后,有47个位点符合哈迪-温伯格平衡(HWE),各位点间未检测到连锁不平衡现象,仅2个位点偏离HWE。结果表明,所开发的大部分微卫星标记具有高多态性,蕴含的遗传信息含量较为丰富,能够为黑棘鲷的种群遗传资源评估提供数量充足、类型多样的有效分子标记。跨物种扩增结果显示,有43个黑棘鲷微卫星标记可在9种鲷科鱼类中成功扩增,其中28个标记在太平洋棘鲷(Acanthopagruspacificus)、黄鳍棘鲷(Acanthopagruslatus)和澳洲棘鲷(Acanthopagrusaustralis)中具有较好的通用性,2个标记在平鲷(Rhabdosargussarba)、蓝点赤鲷(Pagruscaeruleostictus)、真赤鲷(Pagrusmajor)、二长棘犁齿鲷(Evynnis cardinalis)及黄牙鲷(Dentex hypselosomus)中具有通用性。这些通用性标记可为阐明鲷科属、种间的系统进化关系和棘鲷属鱼类的群体遗传学分析提供新的标记来源和研究角度。  相似文献   

东北亚地区M_S6.0以上地震异常电磁波动研究     

杨牧萍  钱庚  张学民  申旭辉  张萌  金艳铭 《大地测量与地球动力学》2022,42(7):669-674

利用2005-01~2009-12 DEMETER电磁卫星详查模式下磁场与电场的极低频（ELF）三分量波形数据,选取东北亚地区（38°~58°N,105°~145°E）6个M_S≥6地震,对震前30 d震中上空±10°的6 000余条1130和1135轨道数据进行波矢分析和坡印廷矢量分析。结果发现,2005-08-16日本本洲东岸近海M_S7.2地震和2009-06-05日本北海道地区M_S6.6地震震前在质子回旋频率以下记录到来源于地球的异常电磁波扰动;2005-11-10俄罗斯西伯利亚东南部M_S6.4地震和2007-08-02俄罗斯萨哈林岛(库页岛)M_S6.8地震震前在质子回旋频率以下没有记录到异常电磁波扰动;2007-02-17日本北海道地区M_S6.2地震和2008-06-13日本本州东岸近海M_S7.3地震震前在质子回旋频率以下记录到异常电磁波扰动,但并非来源于地球。  相似文献   

Genetic variability of half-smooth tongue sole Cynoglossus semilaevis populations using microsatellite markers     

LIU Yunguo  GUO Yinghui  LIU Shuaishuai  LI Junfeng  YE Naihao 《海洋学报(英文版)》2011,30(3):76-83

Ten highly variable microsatellite loci were performed in order to evaluate the genetic variability of wild and hatchery samples of half-smooth tongue sole Cynoglossus semilaevis. A group of 200 genotypes belonging to four wild samples, Laizhou (LZ), Weihai (WH), Qingdao (QD), Rizhao (RZ) and one hatchery sample, Mingbo (MB), were screened. All of the ten microsatellite loci screened in this study showed marked polymorphism. A total of 70 different alleles were observed over all loci. The number of alleles per locus ranged from 4.2 to 10.6. The average of expected and observed heterozygosity ranged from 0.67 to 0.82, and from 0.77 to 0.87, respectively. A total of 10, 16, 10, 11 and 5 unique alleles each were found in LZ, WH, QD, RZ and MB populations. The effective number of alleles varied from 2.87 for HSTS_7 to 6.83 for HSTS-h. The number of average genotypes ranged from 6.0 for HSTS_a to 15.4 for HSTS_h. As compared with the wild populations, the hatchery population, MB, showed significant genetic changes such as fewer alleles per locus (P <0.05), a smaller number of low frequency alleles (P <0.05), a small number of unique alleles and a small number of genotypes (P <0.05), all indicative of a reduction in genetic diversity.  相似文献   

孟加拉笛鲷与四带笛鲷的微卫星分析     

李培  王中铎  郭昱嵩  刘丽  刘楚吾 《台湾海峡》2011,30(4):517-521

采用聚丙烯酰胺凝胶电泳技术,从西大西洋笛鲷（Lutjanuscampechanus）和勒氏笛鲷（L．russel—lii）的微卫星引物中筛选出适用于孟加拉笛鲷（L．bengalensis）和四带笛鲷（三．kasmira）的微卫星引物,并应用于三亚和湛江群体的遗传结构分析．结果表明：25对引物中筛选出13对可以稳定扩增出特异片段的引物,其中12对可在孟加拉笛鲷基因组中扩增出重复性好的特异性条带;10对可在四带笛鲷中扩增出重复性好的特异性条带,并且全部呈现出种内多态;9对为两个种通用．孟加拉笛鲷的湛江群体（zMJ）和三亚群体（SMJ）、四带笛鲷的湛江群体（ZSD）和三亚群体（SSD）的平均等位基因数分别为：3．8889、4．1111、4．3333、4．2222;平均观测杂合度分别为：0．5833、0．6389、0．5944、0．6389;平均多态信息含量分别为：0．4720、0．4692、0．5474、0．5257．将本实验所得结果与笛鲷鱼其他研究结果进行比较分析,可以看出目前湛江海域野生笛鲷的遗传多样性整体比较丰富,但部分野生笛鲷已表现出杂合子缺失的趋势,这势必会引起将来遗传多样性的降低,所以要尽旱对野生群体进行摸底调查。并找出解决养殖群体污染野生群体这一问题的方法．  相似文献   

大菱鲆亲子鉴定的微卫星多重PCR技术建立及应用     

苗贵东  杜民  杨景峰  徐营  樊廷俊  陈松林 《中国海洋大学学报(自然科学版)》2011,(Z1):97-106

用8个微卫星标记组合建立了2个微卫星多重PCR体系,对大菱鲆7个人工选育家系进行了系谱认证、亲子鉴定和遗传多样性研究。2个多重PCR体系中8个微卫星位点共检测到等位基因49个,每个位点等位基因数为3~8个。根据已知亲本及子代基因型,成功推断出了3个缺失亲本的基因型。在双亲未知的情况下2个多重PCR的8个微卫星位点累积排除概率是96.58%,已知1个亲本时累积排除率为99.71%,亲子鉴定准确率为96.42%。采用70个个体进行双盲验证,利用UPGMA法对7个家系的70个体进行了聚类分析,同一家系95.71%的个体聚类分析结果与系谱来源一致。Cervus 2.0软件亲子鉴定结果表明亲子鉴定准确率为92.86%。2个多重PCR体系的建立为大菱鲆不同家系混养后的亲子鉴定、系谱分析和分子辅助家系管理提供了技术手段。  相似文献   

刺参雌核发育二倍体的诱导及其早期生长发育     

聂鸿涛  李琪  于瑞海 《中国海洋大学学报(自然科学版)》2011,41(9):31-35

紫外线照射使精子遗传物质失活,用细胞松弛素B(CB)处理受精卵抑制第二极体释放,诱导刺参(Apostichopusjaponicus)雌核发育二倍体。用强度为2561μW/(cm2.s)的紫外线(254 nm)照射不同时间的精子与正常卵子受精。结果发现随照射时间的增加,卵裂率、早期胚胎存活率和小耳幼虫发生率逐渐降低,照射30 s时小耳幼虫发生率降为0。受精后35 min,显微镜下观察到有20%~30%受精卵排出第1极体时,用浓度0.5μg/mL的CB持续处理受精卵20 min,诱导出刺参第二极体抑制型雌核发育二倍体,雌核发育二倍体发育速度低于正常二倍体。微卫星分析表明,雌核发育二倍体诱导成功率为93.3%。  相似文献   

中国鲎基因组微卫星富集文库的构建及分析     

宁晔凤  黎中宝  戴刚  上官静波  李清荟 《海洋科学》2015,39(4):15-20

本研究旨在从DNA分子水平上研究中国鲎(Tachypleus tridentatus)种群遗传多样性及种群结构等相关信息,以期为保护其野生群体种质资源提供科学依据。本研究通过生物素—磁珠富集法筛选微卫星标记,构建中国鲎基因组微卫星文库。基因组DNA经Fast Digest Tru1I酶切后,选取400 bp~1000 bp片段,用生物素标记的(GT)15、(CT)15混合探针与其杂交,杂交复合物与链霉亲和素磁珠结合,捕获含有重复序列的微卫星片段,纯化后连接PMD19-T载体克隆,构建基因组微卫星文库。从334个阳性克隆中随机选取196个片段大于400 bp的阳性克隆进行测序,共获得127个微卫星序列,其中完美型占69.3%、非完美型占11.0%、复合型占19.7%。除探针使用的GT和CT的重复序列外,还筛选到多碱基GCT、TGG、AAAC、ACAA、GATTT、TTTTA的重复序列。根据微卫星序列设计选择合成40对引物,结果显示其中3对具有较高多态性,可作为进一步评价中国鲎野生种质资源等遗传信息的有效遗传标记。  相似文献   

Estimated reproductive success of brooders and heritability of growth traits for large yellow croaker (Larimichthys crocea) using microsatellites     

���͵�  �����  ��־��  ������  Ҧ���  ���쿭 《中国海洋湖沼学报》2011,29(5):990-995

We estimated the reproductive success of adult brood fish and the heritability of growth-related traits for large yellow croaker (Larimichthys crocea). We created two independent full-factorial cross groups (1 and 2) by crossing 4 males×4 females and 4 males×3 females, respectively. We measured the body weight (BW), body length (BL), and body height (BH) of 281 individuals from group 1 and 318 individuals from group 2 at 20 months post hatch (harvest age). We also collected a tissue sample from each individual. The parents and offspring were genotyped using six polymorphic microsatellites. Of the 599 offspring, 99.2% were assigned to a single pair of parents. In both groups, some parent pairs produced a large number of offsprings while other pairs did not produce any offspring. The genetic diversity and putative Ne were lower in the offsprings than in the parents in both groups. The heritability estimates at 20-month age were 0.13±0.10 for BW, 0.19±0.13 for BL, and 0.09±0.06 for BH. The genetic and phenotype correlation between BW, BL, and BH was close to 1. The results provide basic information for selective breeding and further genetic characterization of large yellow croaker.  相似文献   

Identification and characterization of 43 microsatellite markers derived from expressed sequence tags of the sea cucumber (Apostichopus japonicus)     

Qun Jiang  Qi Li  Hong Yu  Lingfeng Kong 《中国海洋大学学报(英文版)》2011,10(2):185-190

The sea cucumber Apostichopus japonicus is a commercially and ecologically important species in China. A total of 3056 potential unigenes were generated after assembling 7597 A. japonicus expressed sequence tags (ESTs) downloaded from Gen-Bank. Two hundred and fifty microsatellite-containing ESTs (8.18%) and 299 simple sequence repeats (SSRs) were detected. The average density of SSRs was 1 per 7.403 kb of EST after redundancy elimination. Di-nucleotide repeat motifs appeared to be the most abundant type with a percentage of 69.90%. Of the 126 primer pairs designed, 90 amplified the expected products and 43 showed polymorphism in 30 individuals tested. The number of alleles per locus ranged from 2 to 26 with an average of 7.0 alleles, and the observed and expected heterozygosities varied from 0.067 to 1.000 and from 0.066 to 0.959, respectively. These new EST-derived microsatellite markers would provide sufficient polymorphism for population genetic studies and genome mapping of this sea cucumber species.  相似文献   

Development of 57 novel polymorphic microsatellite markers in half-smooth tongue sole (Cynoglossus semilaevis)     

Guidong Miao  Ying Xu  Di Wang  Songlin Chen  Tingjun Fan  Yongsheng Tian 《中国海洋大学学报(英文版)》2011,10(3):241-245

Half-smooth tongue sole (Cynoglossus semilaevis) is a promising species for aquaculture in China.The wild population of C. semilaevis is under threat from environmental factors. Microsatellite markers are very suitable for assessing genetic diversity. Four microsatellite-enriched libraries of half smooth tongue sole (Cynoglossus semilaevis) were constructed,from which 57 polymorphic microsatellites were isolated and characterized.The polymorphism of these microsatellites was assessed by genotyping in 30 ind...  相似文献