基于SLAF-seq技术开发蓝圆鲹微卫星标记及跨物种扩增检测

采用基于高通量测序平台的SLAF-seq技术开发蓝圆鲹(Decapterus maruadsi)微卫星标记,从获得的1 450个四至六碱基重复微卫星位点中随机挑选93个位点进行引物合成和多态性检测,最终开发出31个多态性微卫星标记.群体遗传学检测结果表明,31个位点的等位基因数为4~29(均值13.48),观测杂合度(Ho)为0.281~0.875(均值0.650),期望杂合度(He)为0.557~0.958(均值0.838),多态信息含量值(PIC)为0.456~0.939(均值0.803),表明所开发的31个微卫星位点均具有较高的多态性.Hardy-Weinberg平衡检测表明,17个位点符合HWE("哈迪-温伯格"平衡),且各位点间不存在连锁不平衡现象.跨物种扩增结果显示,分别有27、20、20、18、9个微卫星标记在颌圆鲹(Decapterus macarellus)、长身圆鲹(Decapterus macrosoma)、无斑圆鲹(Decapterus kurroides)、竹荚鱼(Trachurus japonicus)及金带细鲹(Selaroides leptolepis)中具有较好的通用性.本研究开发的多态性长碱基重复微卫星标记可为蓝圆鲹渔业资源的科学管理和保护提供有力的遗传学评估手段,亦可为圆鲹属和鲹科鱼类系统进化研究提供新的分析手段.     

Methods Comparison for Microsatellite Marker Development： Different Isolation Methods, Different Yield Efficiency

Microsatellite markers have become one kind of the most important molecular tools used in various researches. A large number of microsatellite markers are required for the whole genome survey in the fields of molecular ecology, quantitative genetics and genomics. Therefore, it is extremely necessary to select several versatile, low-cost, efficient and time- and labor-saving methods to develop a large panel of microsatellite markers. In this study, we used Zhikong scallop (Chlamys farreri) as the target species to compare the efficiency of the five methods derived from three strategies for microsatellite marker development. The results showed that the strategy of constructing small insert genomic DNA library resulted in poor efficiency, while the microsatellite-enriched strategy highly improved the isolation efficiency. Although the mining public database strategy is time- and cost-saving, it is difficult to obtain a large number of microsatellite markers, mainly due to the limited sequence data of non-model species deposited in public databases. Based on the results in this study, we recommend two methods, microsatellite-enriched library construction method and FIASCO-colony hybridization method, for large-scale microsatellite marker development. Both methods were derived from the mi-crosatellite-enriched strategy. The experimental results obtained from Zhikong scallop also provide the reference for microsatellite marker development in other species with large genomes.     

Inheritance Pattern of Microsatellite Loci and Their Use for Kinship Analysis in the Japanese Scallop Patinopecten yessoensis

The inheritance mode of seven microsatellite markers was investigated in Patinopecten yessoensis larvae from four con-trolled crosses,and the feasibility of using these markers for kinship estimation was also examined. All the seven microsatellite loci were compatible with Mendelian inheritance. Neither sex-linked barriers to transmission nor major barriers to fertilization between gametes from the parents were evident. Two of the seven loci showed the presence of null alleles in two families,suggesting the...     

ISSR 标记在牙鲆(Paralichthys olivaceus)与石鲽(Kareius bicoloratus)种间杂交一代的分离方式分析

利用ISSR标记对牙鲆♀与石鲽♂单对杂交亲本及其子一代(全同胞家系)的分离方式进行了研究。结果表明,8个ISSR引物共扩增出181个标记,其中116个为多态性,占总数的64.09%;标记在F1代呈三种分离方式:符合孟德尔分离比例、偏离孟德尔分离比例和异常分离。符合孟德尔分离比例的情况包括:不分离(代表了亲本基因型的5种组合:AA×AA、AA×Aa、Aa×AA、AA×aa、aa×AA);1︰1分离(代表了亲本基因型的2种组合:Aa×aa、aa×Aa);3︰1分离(代表了亲本基因型的1种组合:Aa×Aa)。偏分离的标记包括:亲本中呈多态而在子代中偏离1︰1分离的标记和亲本均有而在子代中偏离3︰1分离的标记。异常分离的标记包括:F1代出现双亲均不具备的标记和双亲或单亲有而子代却无的标记。3种分离位点出现的频率和数量分别为:82.87%、150,11.05%、20,6.08%、11;双亲共有标记中85.33%的不发生分离,单亲特有标记中28.57%—35.85%发生正常分离,该研究结果可为进一步利用该群体构建牙鲆和石鲽遗传连锁图谱奠定良好的工作基础并提供一定的理论依据。     

牡蛎的遗传育种研究进展

牡蛎肉味鲜美,营养丰富,是世界重要海水贝类养殖对象之一.随着各种生物技术在水产领域的广泛应用,牡蛎的遗传育种工作也备受重视,尤其自Stanley[1]首次人工诱导美洲牡蛎(Crassostrea virginica)三倍体成功以来,大大加快了对牡蛎的遗传学背景和育种进程的研究步伐.笔者就牡蛎遗传育种研究的现状综述如下:     

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。     

长蛸Octopus variabilis自然群体生化遗传学研究

采用同工酶电泳技术,筛选了天冬氨酸转氨酶(AAT)、腺苷酸激酶(AK)、肌酸激酶(CK)、果糖二磷酸激酶(FBP)、3-磷酸甘油醛脱氢酶(G3PDH)、6-磷酸葡萄糖异构酶(GPI)、谷胱甘肽还原酶(GRS)、异柠檬酸脱氢酶(IDH)、甘露糖-6-磷酸异构酶(MPI)和核苷磷酸化酶(NP)等10种同工酶,并以短蛸Octopus ocellatus和目乌贼Sepia aculeata为外群,研究了长蛸Octopus variabilis大连(DLC)、烟台(YTC)和莱州(LZC)3个自然群体的遗传结构和多样性水平.结果表明DLC、YTC、LZC 3个长蛸群体的多态位点比例较高,分别为20%,30%和30%(P_(0.99));群体平均杂合度观测值分别为0.014,0.153,0.092;平均有效等位基因数分别为1.2,1.4,1.3.烟台长蛸群体的遗传多样性较高.对3个种5个群体构建UPGMA系统树,并进行聚类分析.     

南阳和辽河特高含蜡盆地原油中生物标记烃类和油源对比

基于原油中生物标志物的分布，查明了大民屯都是同源成熟油，而南阳则有低成熟油存在，后者可能生于门限深度附近，且未被源岩进一步降解产物昕稀释；基于南阳不同成熟度原油的油源对比效果指出，利用甾烷系列的综台指纹较萜烷更佳。作者认为南阳三种原油成熟度低于相应源岩，可能是由于源岩与储层在块断中的差异沉降，从而引起差异熟化的结果。     

网页制作中元信息标记的应用

介绍了网页制作中元信息标记<META>的功能,以及利用<META>实现页面自动刷新、自动跳转的技巧.