鹿线粒体DNA的种属特异性片段研究

根据对不同产地梅花鹿、马鹿、水鹿、白唇鹿线粒体DNA进行PCR扩增和序列测定,并与亲缘关系较近的伪充药材来源动物线粒体DNA同位置序列比较,找到鹿的特征片段,经过实际检测,筛选出鹿的种属特异性片段。

云南保山猪线粒体DNA D loop区序列初步分析

为了解云南保山猪(Baoshan pig)的遗传多样性及其遗传背景，我们测定了19个个体线粒体DNA Dloop高变区1 1 5 363 - 1 5 801片段序列438帅。检测到1。种单倍型，包括8个多态位点，其中5次T/ C转换、1次G/ A转换、1次G/ C颠换和1次A/ T颠换，其A.T.GX碱基的平均含量分别为35.4%.26.9%.13.2%和24.5 %,A+ T含量(62 .3)明显高于G+ C含量(37 .7 %)。对于保山猪的保种及其持续利用有着重要的理论指导意义。

随机扩增多态DNA影响因素的研究

随机扩增多态 DNA(PAPD）分析受诸多因素的影响, 作者发现不同厂家制造的 PCR 扩增仪, 不同厂家出品的 TaqDNA 聚合酶和 PCR 缓冲液, PAPD 反应体系中的引物浓度, Mg~(2+)浓度, dNTP 浓度, BSA（牛血清白蛋白）和明胶, 以及模板 DNA的量等均可能对 PAPD 结果有不同程度的影响. 为了使 PAPD 结果在不同实验室间具有重复性和可比性, 提高 PAPD 数据的科学价值, 作者建议在 PAPD 分析过程和文章写作中应规范化. 方法部分, 除一般写明的引物浓度、TaqDNA 聚合酶单位数、dNTP 浓度、每一次 PCR 反应的循环条件和循环数等外, 还应注明 PCR 仪的制造厂家及型号、TaqDNA 聚合酶生产厂家 、PCR 缓冲液的成分、Mg~(2+)浓度, 模板 DNA 浓度等。

四川白鹅朗德鹅及杂交后代线粒体DNA多态研究

采用RFLP技术,对四川白鹅和朗德鹅及其杂交后代进行了线粒体DNA(mtDNA)多态分析。在所使用的19种限制性内切酶中,有4个酶(Eco RV、HaeII、HincII和KpnI)检测出多态,共获得两种mtDNA单倍型,四川白鹅和以四川白鹅为母本杂交后代的mtDNA为I型,朗德鹅的mtDNA为II型。以上结果为中国鹅和欧洲鹅存在两种不同起源学说提供了分子遗传学的证据。

云南4个马品种的随机扩增多态DNA(RAPD)分析

腾冲马和新丽江马品种内的遗传变异显著高于文山马和乌蒙马,遗传变异68.06%存在于群体内。系统聚类图中品种内大多数个体聚在一起,文山马和乌蒙马聚成一个大类,腾冲马和新丽江马又聚成一个大类,说明文山马和乌蒙马以及腾冲马和新丽江马有较近的亲缘关系。

云南地方猪种线粒体DNA多态性研究

用18种限制性内切酶分析云南猪种的mtDNA多态性。在全部18头个体中只检出一种限制性类型,结果表明,云南猪种的mtDNA变异度很低,遣传多样性贫乏。

鹿属动物线粒体DNA序列测定的研究

对9种鹿属动物的线粒体DNA进行引物扩增和序列测定,并与牛科、马科的DNA序列进行比较。找到了鹿属动物特异的DNA序列。利用该序列可对鹿类药材进行真伪区分。

东方蜜蜂DNA随机扩增多态及其遗传分化研究

该研究对云南及马来西亚的37个东方蜜蜂样本进行随机扩增多态DNA分析,从20个引物中筛选出11个引物,其中9个引物扩增出多态带。共检测到66条扩增片段,其中56条为多态带。用UPGMA聚类方法构建的分子系统树显示,云南的样本、马来西亚的样本各自分别聚在一起,说明两个样本间遗传差异较大,群体之间存在着遗传分化。但就云南的32个个体而言,虽然聚类图中大多数采自同一地区的样本聚在一起,但也存在一定交叉,提示云南地理群体间近期可能存在一定的基因流。

PHYLOGENY OF RHESUS-MONKEYS (MACACA-MULATTA) AS REVEALED BY MITOCHONDRIAL-DNA RESTRICTION ENZYME ANALYSIS

Mitochondrial DNA, purified from 36 samples of 23 local populations which are widely distributed in Vietnam, Burma, and 10 provinces of China, has been analyzed to model the phylogeny of rhesus monkeys. The 20 local populations of China may represent nearly all major populations in China. Using 20 restriction endonucleases of 6-bp recognition, we observed a total of 50-61 sites in the various samples. By combining the cleavage patterns for each enzyme, the 36 samples were classified into 23 restriction types, each of which was found exclusively in the respective population from which samples were obtained By combining the earlier study of Indian rhesus monkeys, phylogenetic trees, which have been constructed on the basis of genetic distance, indicate that rhesus monkeys in China, Vietnam, India, and Burma can be divided into seven groups. Integrating morphological and geographical data, we suggest that rhesus monkeys in China, Vietnam, and Burma may be classified into six subspecies-M. m. mulatta, M. m. brevicaudus, M. m. lasiotis, M. m. littoralis, M. m. vestita, and M. m. tcheliensis-and rhesus monkeys in India may be another valid subspecies. M. m. tcheliensis is the most endangered subspecies in China. Divergence among subspecies may have begun 0.9-1.6 Ma. The radiation of rhesus monkeys in China may have spread from the southwest toward the east. The taxonomic status of the Hainan monkey and the Taiwan monkey require further investigation.

PHYLOGENETIC-RELATIONSHIPS OF MACAQUES AS INFERRED FROM RESTRICTION-ENDONUCLEASE ANALYSIS OF MITOCHONDRIAL-DNA

Mitochondrial DNAs (mtDNAs) purified from 25 samples of 6 species of macaques, Macaca mulatta, M. fascicularis, M. arctoides, M. nemestrina, M. assamensis and M. thibetana, were analyzed to study the phyletic relationships among the species. A total of 36-46 sites was observed in each sample. By combining the cleavage patterns for each of the endonucleases, the 25 samples were classified into 11 restriction types. When data on M. fuscata and M. cyclopis collected by other authors were added to our own, the resultant molecular phylogenetic trees indicated that the 8 species may be divided into 4 groups: (1) M. mulatta, M. fuscata, M. cyclopis and M. fascicularis; (2) M. arctoides, (3) M. nemestrina; (4) M. assamensis and M. thibetana. Our results suggest that within both the fascicularis and sinica groups genetic distances are small between members and that the status of the species within the groups may require further investigation.

PHYLOGENY OF THE SLOW LORISES (GENUS NYCTICEBUS) - AN APPROACH USING MITOCHONDRIAL-DNA RESTRICTION ENZYME ANALYSIS

Mitochondrial DNA polymorphisms in 15 specimens of three species of slow lorises-Nycticebus coucang, N. intermedius, and N. pygmaeus-were analyzed in order to study the evolutionary relationships among the species. Eight restriction types were observed in the samples. Phylogenetic trees constructed on the basis of genetic distances showed that the slow lorises sort into two clusters: four types of N. coucang and three types of N. intermedius plus one type of N. pygmaeus. Our results suggest that there are two valid species in the genus Nycticebus-N. coucang and N. pygmaeus-and that N. intermedius should be included within N. pygmaeus. Divergence between the two species may have begun 2.7 Ma (million years ago). Evolution of gross morphology, chromosomes, and mitochondrial DNA in the slow lorises appears to be concordant.

DIFFERENT RATES OF MITOCHONDRIAL-DNA SEQUENCE EVOLUTION IN KIRK DIK-DIK (MADOQUA-KIRKII) POPULATIONS

We have investigated evolutionary rates of the mitochondrial genome among individuals of Madoqua kirkii using the relative rate test. Our results demonstrate that individuals of two chromosome races, East African cytotype A and Southwest African cytotype D, evolve about 2.3 times faster than East African cytotype B. Cytogenetic changes, DNA repair efficiency, mutagens, and more likely, hitherto unrecognized factors will account for the rate difference we have observed. Our results suggest additional caution when using molecular clocks in the estimation of divergence time, even within lineages of closely related taxa. Rate heterogeneity in microevolutionary timescales represents a potentially important aspect of basic evolutionary processes and may provide additional insights into factors which affect genome evolution. (C) 1995 Academic Press, Inc.

Phylogenetic relationships of five pika species (genus Ochotona) based on mitochondrial DNA restriction maps

Restriction site mapping of mitochondrial DNA (mtDNA) with 16 restriction endonucleases was used to examine the phylogenetic relationships of Ochotona cansus, O. huangensis, O. thibetana, O. curzoniae and O. erythrotis. A 1-kb length variation between O. erythrotis of subgenus Pika and other four species of subgenus Ochotona was observed, which may be a useful genetic marker for identifying the two subgenera. The phylogenetic tree constructed using PAUP based on 61 phylogenetically informative sites suggests that O. erythrotis diverged first, followed by O. cansus, while O. curzoniae and O. huangensis are sister taxa related to O. thibetana, The results indicate that both O. cansus and O. huangensis should be treated as independent species. If the base substitution rate of pikas mtDNA was 2% per million years, then the divergence time of the two subgenera, Pika and Ochotana, is about 8.8 Ma ago of late Miocence, middle Bao-dian of Chinese mammalian age, and the divergence of the four species in subgenus Ochotona would have occurred about 2.5 - 4.2 Ma ago, Yushean of Chinese mammalian age. This calculation appears to be substantiated by the fossil record.

Extraction, PCR amplification, and sequencing of mitochondrial DNA from scent mark and feces in the giant panda

To expand the feasibility of applying simple, efficient, non-invasive DNA preparation methods using samples that can be obtained from giant pandas living in the wild, we investigated the use of scent markings and fecal samples. Giant panda-specific oligonucleotide primers were used to amplify a portion of the mitochondrial DNA control region as well as a portion of the mitochondrial DNA cytochrome b gene and tRNA(Thr) gene region. A 196 base pair (bp) fragment in the control region and a 449 bp fragment in the cytochrome b gene and tRNA(Thr) gene were successfully amplified. Sequencing of polymerase chain reaction (PCR) products demonstrated that the two fragments are giant panda sequences. Furthermore, under simulated field conditions we found that DNA can be extracted from fecal samples aged as long as 3 months. Our results suggest that the scent mark and fecal samples are simple, efficient, and easily prepared DNA sources. (C) 1998 Wiley-Liss, Inc.