Microsatellite DNA polymorphisms and the relation with body weight in sea cucumber Apostichopus japonicus

The relationship between microsatellite polymorphism and body weight of captive bred Chinese sea cucumber Apostichopus japonicus was investigated in two local populations in Dalian.Among ten loci discovered, nine show changes except for A J07 loci. Seven loci were found highly polymorphic in both populations. For each locus in two populations, the average number of alleles is 6.428 6 and 6.285 7, the average observed heterozygosity at 0.225 7 and 0.245 9, the expected heterozygosity at 0.776 8 and 0.748 8, the polymorphism information content (PIC) at 0.709 2 and 0.674 6, respectively. Further analysis show significant correlation between A. japonicus body weight and occurrence markers AJ02 and AJ04. The findings of the relation may be helpful for molecular breeding,as well as the marker-assisted selection of sea cucumbers.     

Juvenile fish condition in estuarine nurseries along the Portuguese coast

Connectivity between estuarine fish nurseries and coastal adult habitats can be affected by variations in juvenile growth and survival. Condition indices are renowned proxies of juvenile nutritional status and growth rates and are valuable tools to assess habitat quality. Biochemical (RNA:DNA ratio) and morphometric (Fulton's condition factor K) condition indices were determined in juveniles of Solea solea, Solea senegalensis, Platichthys flesus, Diplodus vulgaris and Dicentrarchus labrax collected in putative nursery areas of nine estuaries along the Portuguese coast (Minho, Douro, Ria de Aveiro, Mondego, Tejo, Sado, Mira, Ria Formosa and Guadiana) in the Spring and Summer of two consecutive years (2005 and 2006) with distinct climatic characteristics. Individual condition showed significant variation amongst species. The combined use of both condition indices highlighted the low correlation between them and that RNA:DNA had a higher sensitivity. RNA:DNA varied between years but overall the site relative patterns in condition were maintained from one year to the other. Higher RNA:DNA values were found in Spring than in Summer in most species. Intra-estuarine variation also occurred in several cases. Species specific trends in the variability of condition amongst estuaries were highlighted. Some estuaries had higher juvenile condition for more than one species but results did not reveal an identical trend for all species and sites, hindering the hypotheses of one estuarine nursery promoting superior growth for all present species. Significant correlations were found between condition indices, juvenile densities and environmental variables (water temperature, salinity and depth) in the estuarine nurseries. These influenced juvenile nutritional condition and growth, contributing to the variability in estuarine nursery habitat quality. Management and conservation wise, interest in multi-species approaches is reinforced as assessments based on a single species may not reflect the overall nursery habitat quality.     

柳珊瑚4种不同DNA提取方法的比较研究

通过2种样品前处理方式和4种不同的DNA提取方法相结合,提取橘色刺柳珊瑚(Echinogoria aurantiaca)基因组DNA,并对其18S rRNA基因进行了克隆和测序。实验结果表明,对于2种样品前处理方式,采用纯酒精进行样品前处理比PBS(磷酸缓冲液)进行样品前处理效果要好。在4种不同的DNA提取方法中,以CTAB(十六烷基三甲基溴化铵)法提取基因组DNA效果最好,其余3种方法如PVP(聚乙烯基吡咯烷酮)法、高盐法和酚/氯仿法的提取效果较差。并且不同样品的钙质沉淀对DNA都有一定程度的吸附。最后,通过PCR扩增、克隆测序得到橘色刺柳珊瑚18S rRNA基因序列(AY962532)。BLAST软件进行序列比对表明橘色刺柳珊瑚与弱柳珊瑚(Leptogorgia chilensis)的18S rRNA基因(AF052928)同源性最高,为98.79%。     

线粒体基因片段在梭子蟹系统发育及物种鉴定中的应用

测定三疣梭子蟹(Portunus trituberculatus)9个个体COI基因部分序列467 bp,7个个体16S rRNA基因部分序列519 bp,同时测定样品蟹1个个体COI基因部分序列468 bp.结合GenBank中收集的梭子蟹科COJ,16S rRNA两个基因所有同源序列信息,使用Kimura双参数模型采用邻接法(NJ)、最大简约法(MP)构建梭子蟹科分子系统发育树;将样品蟹测得的COI基因序列和已知鲟属的其他蟹同源序列(429 bp)进行比对分析.结果分析表明:两个基因片段序列平均碱基AT数量分数都明显高于GC数量分数;梭子蟹属与美青蟹属关系最近,蟳属应为区别于梭子蟹属、美青蟹属、青蟹属的另一支,支持蟳属应划分在短桨蟹亚科的观点;根据遗传距离以及转换/颠换(R)值分析,判定出样品蟳为锐齿蟳.本试验运用线粒体基因片段探讨了梭子蟹类的系统发育关系以及样品蟹的种类鉴定,为线粒体基因片段在梭子蟹的物种鉴定和系统发育重建中的开发和利用提供重要参考.     

大亚湾紫红笛鲷野生群体遗传多样性的RAPD分析

紫红笛鲷野生群体样品取自广东大亚湾海区,取背部肌肉,提取DNA,用RAPD技术检测遗传多样性。40个随机引物当中,有11个引物得到了条带清晰、位点较多且重复性好的电泳图谱。11个引物共得到76个条带,其中48个多态位点,多态位点比例为63.16%。平均遗传距离为0.2144。遗传多样性指数为0.1876。结果显示,大亚湾海区的紫红笛鲷具有较丰富的遗传多样性,工程建设以及人为捕捞等对紫红笛鲷的遗传结构影响不明显。在该海区进行紫红笛鲷的养殖,可以选择野生紫红笛鲷作为亲本培育种苗,但应该注意合理的保护,防止遗传多样性的丧失。     

两种扇贝杂交和自交家系早期生长及甲基化的比较分析

运用杂交和自交方法建立虾夷扇贝♂(A)×栉孔扇贝♀(B)、栉孔扇贝♀(B)×栉孔扇贝♂(C)、栉孔扇贝♂(C)×栉孔扇贝♀(D)3 种组合家系, 每个组合3 个平行, 共9 个家系。对各家系卵径大小、胚胎孵化率、幼虫浮游阶段壳长生长速度等生物学参数进行比较, 建立了幼虫浮游期壳长与日龄的线性回归方程; 运用甲基化敏感扩增多态性(MSAP)分析了基因组DNA 胞嘧啶甲基化水平, SPSS 分析了DNA 甲基化与壳长生长的相关性。结果表明, 杂交家系的卵径大小、胚胎孵化率与自交组没有显著差异, 但壳长生长速度显著高于自交组, 这种优势可能主要来自于父本的影响; 家系AB、BC、CD 的平均DNA甲基化率分别为18.672%、22.661%和22.303%, 杂交使后代DNA 甲基化水平降低, 生长速度与DNA甲基化水平相关系数为?0.934, 相关水平极显著(P<0.01), DNA 甲基化与杂种优势具有一定的关系。     

PCR法制备地高辛标记探针斑点杂交 检测白斑综合症病毒(WSSV)

用PCR法成功制备了DIG标记探针，探针长度为547bp，探针的产量为21.6ng/μL。此探针与随机引物合成探针检测样品灵敏度相近。用此探针核酸斑点杂交法检测了54尾中国对虾。结果表明此探针在对白斑综合症病毒的检测、对虾暴发性流行病的诊断等方面具有很高的应用价值。     

消油剂和120#燃料油对海胆DNA损伤及甲基化影响

利用120#燃料油分散液和消油剂处理的乳化液测定海胆急性毒性和慢性DNA损伤及全基因组甲基化水平变化,确定消油剂处理的120#燃料油对海胆的毒性影响。结果表明:消油剂单独使用对海胆的影响微乎其微,经消油剂处理的燃料油乳化液中的多环芳烃质量浓度明显高于分散液中的质量浓度;乳化液和分散液对海胆都有明显的毒性作用,其96h半数效应浓度(EC50)值分别为12.03g/L和29.15g/L;DNA损伤研究表明,乳化液处理的海胆肠细胞彗星拖尾情况比同质量浓度分散液中拖尾情况更加严重;全基因组甲基化水平比分散液中更低,表明乳化液的急性毒性更大,所造成的DNA损伤更加严重。结果提示消油剂的使用会增大燃料油对海胆的毒性效应。