An examination of spatial and temporal genetic variation in walleye pollock (Theragra chalcogramma) using allozyme, mitochondrial DNA, and microsatellite data

We used allozyme, microsatellite, and mitochondrial DNA (mtDNA) data to test for spatial and interannual genetic diversity in wall-eye pollock (Theragra chalcogramma) from six spawning aggregations representing three geographic regions: Gulf of Alaska, eastern Bering Sea, and eastern Kamchatka. Interpopulation genetic diversity was evident primarily from the mtDNA and two allozyme loci (SOD-2*, MPI*). Permutation tests ˆindicated that FST values for most allozyme and microsatellite loci were not significantly greater than zero. The microsatellite results suggested that high locus polymorphism may not be a reliable indicator of power for detecting population differentiation in walleye pollock. The fact that mtDNA revealed population structure and most nuclear loci did not suggests that the effective size of most walleye pollock populations is large (genetic drift is weak) and migration is a relatively strong homogenizing force. The allozymes and mtDNA provided mostly concordant estimates of patterns of spatial genetic variation. These data showed significant genetic variation between North American and Asian populations. In addition, two spawning aggregations in the Gulf of Alaska, in Prince William Sound, and off Middleton Island, appeared genetically distinct from walleye pollock spawning in the Shelikof Strait and may merit management as a distinct stock. Finally, we found evidence of interannual genetic variation in two of three North American spawning aggregations, similar in magnitude to the spatial variation among North American walleye pol-lock. We suggest that interannual genetic variation in walleye pollock may be indicative of one or more of the following factors: highly variable reproductive success, adult philopatry, source-sink metapopulation structure, and intraannual variation (days) in spawning timing among genetically distinct but spatially identical spawning aggregates.

药蒲公英（Taraxacum officinale weber）耐盐变异体的筛选及其耐盐性分析研究

以药蒲公英（Taraxacum officinale Weber）叶片外植体为材料诱导愈伤组织。以NaCl作为选择因子，从愈伤组织直接筛选。在选择培养基上，大部分愈伤组织褐化死亡，在一些褐化死亡的愈伤组织周围有少量新的细胞团生长，挑选生长存活状况好的细胞团转接到新鲜培养基上，每3周继代一次，经3个月继代筛选获得了耐1.5% NaCl的药蒲公英细胞团。以普通愈伤为对照，发现随着NaCl浓度的升高，耐盐愈伤的相对生长率下降但显著高于对照;且随着盐胁迫处理时间的延长持续升高，而普通愈伤对照几乎停止生长，说明耐盐愈伤具有相对稳定的耐盐性。在蛋白水平上，耐盐愈伤与对照愈伤差异明显，SDS-PAGE分析显示：耐盐愈伤比对照多出一条34 KD大小的蛋白带，且30 KD，18 KD左右的蛋白带明显上调。相同处理条件下耐盐愈伤脯氨酸的增加幅度高于对照。盐胁迫条件下，耐盐愈伤的超氧化物歧化酶（SOD）、过氧化物酶（POD）和过氧化氢酶（CAT）活性明显高于对照，且随着处理时间的延长和盐浓度的增加呈现升高的趋势，而对照则呈现先升高后下降的趋势。1.5% NaCl处理前后，耐盐愈伤的总黄酮含量显著高于对照。结果说明耐盐愈伤一方面通过积累蛋白和其他小分子有机溶质的方式调节其渗透平衡，另一方面还可通过提高抗氧化能力降低盐分造成的次级伤害。 将耐1.5% NaCl的药蒲公英愈伤组织接种在分化培养基上分化出芽，之后将再生芽转接到生根培养基中进行生根培养，经4个月得到了12株耐1.5% NaCl的药蒲公英再生植株。与野生型相比，耐盐植株叶片宽大、叶柄粗短、叶表面覆盖白色细毛，根粗壮较短，花茎中部具有2 cm左右的苞叶。RAPD和SDS-PAGE检测表明，耐盐植株与对照植株在DNA及蛋白水平上均存在明显差异。1.5% NaCl处理后，与普通再生植株相比，耐盐株系的抗氧化酶活性明显提高，脯氨酸含量上升幅度更为显著，而丙二醛含量降低，其主要药用成分黄酮的含量显著增加。这些结果说明耐盐植株的抗氧化防御能力明显增强。以上结果表明耐1.5% NaCl的药蒲公英再生植株为耐1.5% NaCl药蒲公英变异体，这些耐盐变异体有望成为抗盐耐海水蔬菜家族的新成员。同时，这些耐盐变异体植株比普通植株具有更高的医用商业价值。耐1.5% NaCl的药蒲公英再生变异体遗传稳定性的研究正在进行中。

黑麂和费氏麂四种卫星DNA的克隆特征和染色体定位

近年来,分子细胞遗传学研究已基本证实了染色体的串联融合(端粒一着丝粒融合)是麂属动物核型演化的主要重排方式.尽管染色体串联融合的分子机制还不清楚,但通过染色体的非同源重组,着丝粒区域的卫星DNA被认为可能介导了染色体的融合.以前的研究发现在赤麂和小麂染色体的大部分假定的串联融合位点处存在着非随机分布的卫星DNA.然而在麂属的其他物种中,这些卫星DNA的组成以及在基因组中的分布情况尚未被研究.本研究从黑麂和费氏麂基因组中成功地克隆了4种卫星DNA (BMC5、BM700、BM1.1k和FM700),并分析了这些卫星克隆的特征以及在小麂、黑麂、贡山麂和费氏麂染色体上的定位情况.结果表明,卫星Ⅰ和Ⅱ DNA (BMC5,BM700和FM700)的信号除了分布在这些麂属动物染色体的着丝粒区域外,也间隔地分布在这些物种的染色体臂上.其研究结果为黑麂、费氏麂和贡山麂的染色体核型也是从一个2n=70的共同祖先核型通过一系列的串联融合进化而来的假说提供了直接的证据.

云南4个少数民族的随机扩增多态DNA分析

1990年,Williams和Welsh领导的2个小组几乎同时独立地发展起来一项新技术,即随机扩增多态DNA(Random amplified polymorphic DNA,RAPD).该技术通过PCR进行DNA扩增,所用引物是G+C含量为50%—70%的单个随机短引物,这些引物在一定的退火条件下能与基因组DNA中的互补顺序配对,启动DNA的合成.RAPD具有以下特点:(1)无需预先知道受试有机体基因组DNA的序列,因而能应用于所用的生物体;(2)绝大多数

滇金丝猴的随机扩增多态DNA与遗传多样性分析

对6只笼养滇金丝猴(Rhinopithecus bieti)进行了随机扩增多态DNA(RAPD)及遗传多样性分析.用45个10bp随机短引物对每只滇金丝猴的基因组DNA进行了扩增,平均每个个体观察到的RAPD标记约为130个左右,单个引物获得的标记在1～7个之间.80%的RAPD标记表现为无多态的单型性.个体间的遗传距离为0.052,表明笼养滇金丝猴群体的遗传多样性很低.此研究结果与在蛋白多态研究中得到的一致.贫乏的遗传多样性一方面使目前处于濒危境地的滇金丝猴生存情况更加危险,同时其本身也可能是造成目前滇金丝猴濒危的原因之一.另外,通过成对的遗传距离分析,构建了这一群滇金丝猴的谱系关系图,提出了让遗传距离较远的个体间进行交配的笼养繁育计划.

中国黑冠长臂猿的遗传多样性及其分子系统学研究--非损伤取样DNA序列分析

采用非损伤性DNA基因分型技术(Noninvasive DNA genotyping),对我国珍稀灵长类动物黑冠长臂猿11个个体的线粒体DNA(mtDNA)控制区159bp的片段进行了序列分析。根据分子系统树,结合形态学方面的资料,提出对中国黑冠长臂猿新的分类观点,即现生的中国黑冠长臂猿应为3个种(H. concalor; H. leucogenys; H. hainanus),其中 H. concolor含3个亚种(H.c.concolor, H.c.jingdongensis, H.c.furvogaster)。同时,针对该类珍稀动物保护,提出将上述黑冠长臂猿的种和亚种作为不同的进化显著性单元(Evolutionarilly Sigificant Units, ESU)进行保护和遗传管理。

鹿鞭的微量DNA提取及序列鉴定

采用微量DNA提取技术,从梅花鹿血、毛、鹿鞭、鹿茸、牛鞭、驴鞭中提取DNA,以线粒体DNA细胞色素b通用引物L14841和H15149扩增约307bpDNA片段, 扩增产物纯化后采用双脱氧链终止法测定其序列。结果证明:梅花鹿毛、血和鹿鞭的DNA序列完全一致; 而所谓的“鹿茸”则与其有较大的差异。用所测序列以简约法PAUP3.1.1 程序构建的分子系统树与传统分类系统相吻合。

云南绵羊线粒体DNA遗传多态性研究

该研究用12种识别六碱基的限制性内切酶对来自云南省3个地方绵羊品种11个样本的线粒体DNA(mtDNA)进行了限制性片段长度多态性分析(RFLP)，结果表明，云南绵羊群体平均核苷歧异度为0.086%，平均遗传距离为0.004，说明mtDNA变异度很低，遗传多样性贫乏，提示云南绵羊可能起源于同一个共同祖先;昭通绵羊与德钦绵羊距离较远，大约分歧于30万年前，德钦绵羊与腾冲绵羊大约分歧于11万年前，而腾冲绵羊与昭通绵羊也于18万年前发生分歧。

安徽麝线粒体DNA细胞色素b基因全长序列分析

对安徽麝模式皮张标本进行了线粒体DNA细胞色素b基因全长序列分析。研究结果表明,安徽麝同麝属中其他种的遗传分化已经相当明显。分子系统学的分析表明,安徽麝是一个单系群,它同麝属其他种的DNA序列差异已达到种间分化的程度。因此,线粒体DNA序列的证据支持将安徽麝列为麝属中一有效种(Moschus anhuiensis),而不是前人认为的原麝或林麝的亚种。

MULTIPLE GENOTYPES OF MITOCHONDRIAL-DNA WITHIN A HORSE POPULATION FROM A SMALL REGION IN YUNNAN PROVINCE OF CHINA

mtDNA genotypes of six domestic horses (three adult short horses whose heights are under 1 m and three common domestic horses) from a small region of 15 km(2) in Malipo county of Yunnan province of China were investigated by the technique of restriction fragment length polymorphism (RFLP) with restriction endonucleases which recognize 6-bp sequences. An average of fragments for an individual was obtained. Unlike other domestic animals, this population of horses exhibits high mtDNA genetic diversity. Each of the six horses has a specific mtDNA genotype showing a pattern of multiple maternal origins, as suggested by fossil and literature records. We think the population of horses is an amazing seed-resource pool of horses and hence deserves to be paid more attention from the view of conservation genetics. However it is also remarkable that we did not find any typical mtDNA genetic markers which would discriminate between short horses and common domestic horses.

Genetic diversity and population history of the red panda (Ailurus fulgens) as inferred from mitochondrial DNA sequence variations

The red panda (Ailurus fulgens) is one of the flagship species in worldwide conservation and is of special interest in evolutionary studies due to its taxonomic uniqueness. We sequenced a 236-bp fragment of the mitochondrial D-loop region in a sample of 5