日本稻蝗部分种群遗传多样性研究

采用随机引物扩增多态性分析方法,研究我国长江以南地理距离跨度较大的6个不同种群日本稻蝗Oxya japonica的种群遗传学关系.10条随机引物对57个日本稻蝗个体共扩增出131个片段,其中多态性位点为128个,总的多态位点百分率是97.7%.种群问的遗传分化系数表明:日本稻蝗的遗传变异大部分存在于种群内.Nei's遗传距离显示:日本稻蝗种群问的遗传距离存在较大差异,河北平山种群和云南种群遗传距离最大,为0.1715,广东广州种群和广西南宁种群遗传距离最小,为0.0770.聚类结果分析表明,地理上相距较近的种群优先相聚,遗传距离、地理距离和亲缘关系间呈现出一定的相关趋势.     

山西省及邻近地区中华稻蝗5个种群RAPD分析及其亲缘关系

采用10条10bp随机引物对来自山西省临猗县(山西南部)、屯留县、太原市近郊(山西中部)、代县(山西北部)、内蒙呼和浩特市郊5个不同种群的中华稻蝗(Oxya chinensis)和广西南宁日本稻蝗(Oxya japonica)共64个个体进行了RAPD分析,探讨了中华稻蝗5个不同种群的亲缘关系。结果表明：10条随机引物对供试的64个个体共产生115条谱带。所获片段的分子量大小为200～2500bp,单个引物扩增的谱带数是5～15条,平均为12条。根据扩增出的RAPD图谱,运用SPSS软件包中的Between-groups linkage法进行聚类分析,聚类结果显示,中华稻蝗不同种群的个体优先聚为1类;供试中华稻蝗5个种群的系统树分为两大分支,山西临猗种群、屯留种群亲缘关系最近,首先相聚,然后与太原种群聚为一支,山西代县和内蒙呼和浩特2种群聚为另一支,两支相聚后与日本稻蝗相聚。表明供试中华稻蝗5个种群存在一定程度的差异,其亲缘关系的远近与地理距离之间存在一定的正相关关系。     

稻蝗属部分种类RAPD及其分子系统学关系

用经筛选的8个随机引物对稻蝗属(Oxya Audient-Serville)中的中华稻蝗(Oxya chinensis)、日本稻蝗(Oxyajaponica)、山稻蝗(0xya agaxisa)、短翅稻蝗(Oxya brachyptera)和小稻蝗(Oxya intricata)等5个种41个个体进行扩增,均扩增出特异性产物,在96条清晰稳定的扩增片段中,多态性片段为95条,种间共有片段1条(引物S362扩增的约850 bp的片段),所获片段的长度大小为200—2500bP,单个引物扩增的条带数在8-16之间,平均为12。根据扩增结果,应用SPSS软件包中的Between-groups linkage法进行聚类分析,得到5种稻蝗的分子系统树。结果显示：中华稻蝗与短翅稻蝗关系密切,日本稻蝗与山稻蝗关系较近,而小稻蝗与它们的亲缘关系较远。聚类图所呈现的种间关系与传统的形态及细胞分类学结果基本一致,5种稻蝗亲缘关系的远近程度得以清楚显示,说明RAPD技术在稻蝗属种间分子系统学研究中具有很好的适用性。     

中华稻蝗不同地理种群遗传多样性的AFLP分析

为研究中华稻蝗种群遗传多样性和遗传结构, 文章对我国7省市的7个中华稻蝗种群进行AFLP分析。选取7对引物扩增128个个体, 共产生336条带, 多态性条带292条, 占86.90%。结果表明: 中华稻蝗种群具有较高的遗传多样性水平, 其中海南万宁种群的遗传多样性高于其他各种群。Mantel检验(r=0.27, P=0.89)表明中华稻蝗各种群遗传距离与地理距离间没有显著相关性。种群间具有明显的遗传分化现象。UPGMA(Unweighted pair group method average)聚类分析显示, 7个中华稻蝗种群按地理距离分为3支: 北方北京昌平、山西太原和山东济宁为一支; 南方陕西汉中、湖南长沙和广西来宾为一支; 海南万宁单独为一支。上述结果以及PCA(Principal component analysis)分析均表明由于地理隔离中华稻蝗种群显示出明显的南北分化和岛屿大陆种群遗传分化现象。     

山稻蝗不同地域和种群染色体C带核型研究

对我国分布的山稻蝗Oxya agavisa Tsai不同种群进行了染色体C带核型研究,并对山稻蝗武夷山种群染色体C带核型进行了深入探讨,分析了其带型特殊性及该种群与其它山稻蝗种群染色体C带核型的区别与联系。同时通过对山稻蝗与中华稻蝗Oxya chinensis(Thunberg)日本稻蝗Oxya japonica(Thunberg）的形态,分布及染色体C带核型等方面的比较,探讨了该3个种之间的进化关系,认为作稻蝗属中的大型种类,上述3个种之间有着较近的亲缘关系。染色体带型的实验结果表明,中华稻蝗为较原始的种类,而日本稻蝗和山稻蝗则可能是由原始中华稻蝗进化而来,其中日本稻蝗L2染色体的形成是由于原始中华稻蝗该染色体常染色质部分异染色质化的结果,山稻蝗L2染色体则可能是由于原始中华稻蝗该染色体近端部的臂间倒位所致。通过对武夷山山稻蝗的研究,对日本稻蝗和山稻蝗之间的近缘关系进行了分析和讨论。     

山稻蝗不同种群的遗传多样性研究

应用RAPD技术对我国长江以南丘陵山区6个种群山稻蝗Oxya agavisa进行了遗传多样性分析.10条随机引物共扩增出90条清晰稳定的片段,其中多态性位点为87个,总的多态位点百分率96.7%.采用Shannon's遗传多样性值和Nei's基因多样性指数分析种群内的遗传多样性及种群间的遗传分化,结果表明山稻蝗湖南通道种群遗传多样性最高;两种指数估算的种群间遗传分化系数(44.5%,41.3%)均表明:山稻蝗种群内的遗传多样性稍大于种群间的遗传多样性.分别用Between-groups linkage、UPGMA和NJ法构建所有个体间的聚类关系图,聚类结果分析表明:位于同一纬度附近且地理上相距较近的种群优先相聚,聚类关系与生境条件、地理距离间呈现出一定的相关趋势.     

利用RAPD对稻蝗属昆虫亲缘关系的研究

通过20个随机引物的PCR扩增,得到了日本主要稻蝗的随机扩增多态性DNA（RAPD）图谱,根据扩增结果,计算了种间相似系数和遗传距离,建立了UPGMA系统树。结果表明,分布没有重叠、种间容易交配、能产生杂种的中华稻蝗台湾亚种与小翅稻蝗的亲缘关系最近;分布重叠的日本稻蝗与中华稻蝗台湾亚种、日本稻蝗和小翅稻蝗的亲缘关系较近。小稻蝗与其它3种稻蝗的亲缘关系较远。     

云南三种稻蝗基因组DNA的RAPD多态性分析

应用RAPD技术对采自云南龙陵两交水和腾冲曲石马鹿冲的24头稻蝗进行了基因组DNA多态性分析,10条随机引物共产生129条清晰、稳定的谱带.基于Jaccard相似系数,用Between-groups Linkage法得出的聚类图与基于Nei's遗传距离分别用UPGMA和NJ法构建的分子系统树基本一致:供试的所有稻蝗个体分为3个聚类簇:龙陵两交水的3、7～8号个体与腾冲曲石马鹿冲(海拔1450m)的8个个体聚为一支,龙陵两交水的1、2、4～6号个体聚为另一支,两支相聚后与腾冲曲石马鹿冲(海拔1 550 m)的8个个体聚为一起.形态学鉴定表明:龙陵两交水的3、7～8号个体为日本稻蝗,1、2、4～6号个体为小稻蝗,腾冲曲石马鹿冲(海拔1450 m)的8个个体均为日本稻蝗,腾冲曲石马鹿冲(海拔1550 m)的个体为有待进一步鉴定的未知种.形态学鉴定结果与聚类结果完全吻合,充分展示了RAPD技术在区分近缘物种方面的独到优势.比较不同采集地的日本稻蝗、小稻蝗的RAPD分析结果,发现云南的这两种稻蝗在同物种不同种群中的遗传多样性最为丰富,从一个侧面显示出我国云南昆虫的物种多样性及物种内部丰富的遗传多样性.     

中华稻蝗三种群遗传结构分析

应用淀粉凝胶电泳对山西省太原黄陵村、临猗县赵村及永济县伍姓湖3个中华稻稻（Oxya chinensis）种群的4个等位酶位点（MDH-1、MDH-2、LDH、ME）进行了分析。结果表明3个种群的中华稻蝗在遗传结构上具有差异;黄陵种群的MDH-1、MDH-2以及伍姓湖种群的MDH-1均为单态位点;赵村种群的杂合性最低（H=0.112）,伍姓湖种群最高（H=0.229）;赵村种群与伍姓湖种群间的Nei‘s遗传距离为0.068,可视为1个大的种群,而黄陵种群与赵村种群和伍姓湖种群的Nei‘s遗传距离分别为0.247和0.218。考虑到赵村和伍姓湖之间的地理距离（41km）比太原黄陵（357km）要近得多,结果提示3个种群地理距离与遗传结构差异之间存在相关关系。     

山稻蝗不同地域种群染色体C带核型研究

对我国分布的山稻蝗OxyaagavisaTsai不同种群进行了染色体C带核型研究 ,并对山稻蝗武夷山种群染色体C带核型进行了深入探讨 ,分析了其带型特殊性及该种群与其它山稻蝗种群染色体C带核型的区别与联系。同时通过对山稻蝗与中华稻蝗Oxyachinensis (Thunberg)、日本稻蝗Oxyajaponica (Thunberg)的形态、分布及染色体C带核型等方面的比较 ,探讨了该 3个种之间的进化关系 ,认为作为稻蝗属中的大型种类 ,上述 3个种之间有着较近的亲缘关系。染色体带型的实验结果表明 ,中华稻蝗为较原始的种类 ,而日本稻蝗和山稻蝗则可能是由原始中华稻蝗进化而来 ,其中日本稻蝗L2染色体的形成是由于原始中华稻蝗该染色体常染色质部分异染色质化的结果 ,山稻蝗L2染色体则可能是由于原始中华稻蝗该染色体近端部的臂间倒位所致。通过对武夷山山稻蝗的研究 ,对日本稻蝗和山稻蝗之间的近缘关系进行了分析和讨论。     

Isolation of 10 tetranucleotide microsatellite loci in the blackcap (Sylvia atricapilla)

We isolated 10 polymorphic microsatellite DNA loci from the blackcap (Sylvia atricapilla) and optimized them for future studies of population differentiation in populations with different migration strategies in southwestern Germany. The loci were screened for polymorphism using 178 individuals from two populations in Germany and Spain. The primers amplified highly variable loci characterized by two to 19 alleles per locus and their observed and expected heterozygosities range from 0.47 to 0.81 and from 0.50 to 0.91, respectively.     

Uniformity of the nuclear and chloroplast genomes of Spartina maritima (Poaceae), a salt-marsh species in decline along the Western European Coast

Spartina maritima is a salt-marsh species from European and African Atlantic coasts. In the northern range of the species (including north-west France), a rapid decline of the populations has been observed during the 20th century. In this paper, the molecular diversity of 10 populations of S. maritima from France has been investigated using nuclear and chloroplast DNA markers: inter-simple sequence polymorphism (ISSR), randomly amplified polymorphic DNA (RAPD), inter-retrotransposon amplified polymorphism (IRAP), and PCR-RFLP of a 5 kb long portion of chloroplast DNA. The results reveal an extremely low level of genetic variation in this species: only one nuclear marker (out of 98) was polymorphic, with the presence of two genotypes randomly distributed among the populations. The lack of genetic diversity is interpreted in light of the almost exclusive vegetative propagation of the species in its northern range, the colonization history of the populations, and the origin of S. maritima (2n = 60) in the hexaploid lineage of the genus and in the context of the management of S. maritima populations.     

Extrachromosomal elements in the lower eukaryote Leishmania

Extrachromosomal DNA elements have been identified in wild-type populations of the parasitic protozoan Leishmania. Elements from L. major and L. tropica were detected using orthogonal-field-alternation-gel electrophoresis. They are nonhomologous, supercoiled circular DNA molecules derived from different chromosomes in the Leishmania genome. Electron microscopy revealed that the elements have very similar physical properties; both are 80-kilobase supercoiled DNA molecules that contain large inverted repeat structures. The extrachromosomal DNAs are amplified in the Leishmania populations and show a fluctuation in copy number, from undetectable to around 20 copies per cell. After exposure of the L. tropica population to the drug methotrexate (MTX), a second amplified DNA was observed that is homologous to the extrachromosomal DNA found in L. major. Furthermore, wild-type Leishmania populations containing extrachromosomal DNA adapt more readily to MTX selection than populations with no amplified DNA. From these observations, there appears to be a relationship between the presence of extrachromosomal elements in wild-type Leishmania and the genesis and maintenance of MTX resistance in these organisms.     

浙江产车前(Plantago asiatica)种群遗传分化的主坐标分析

运用18个有效的10mer-寡核苷酸引物对浙江产车前（Plantago asiatica)的8个种群基因组DNA进行了PCR扩增,共检测到186个位点,其中多态位点125个,占68.70%,应用Jaccard公式计算了8个种群在125个位点上的相似性,以此为基础,应用主坐标排序,作出了8个种群遗传分化的三维排序图,结果表明,8个种群的遗传分化与地理位置、海拔高度有联系。     

Genetic variation of Usnea filipendula (Parmeliaceae) populations in western Germany investigated by RAPDs suggests reinvasion from various sources

Random amplified polymorphic DNA (RAPD) markers were characterized for 25 specimens of Usnea filipendula to evaluate the genetic diversity of populations reinvading formerly uninhabited regions in Northrhine-Westphalia due to decreasing sulfur dioxide levels. With six 10-mer randomly amplified polymorphic DNA (RAPD) primers, a 66 character by 25 specimens matrix was generated. Phenetic analysis (UPGMA) showed no obvious groupings. The reinvading populations are distributed over the phenogram and are not genetically closely related. The results suggest that the reinvading populations of this usually sterile species are derived from different sources and do not consist of a particular clone capable of re-entering the area.     

Genotype relationships inMicroseris elegans (Asteraceae,Lactuceae) revealed by DNA amplification from arbitrary primers (RAPDs)

The genetic relationships among 10 inbred lines representing 10 populations of the autogamous annualMicroseris elegans from throughout California has been determined using random amplified polymorphic DNAs (RAPDs). Seventeen arbitrary 10 base pair primers produced 134 amplification products; 81 of these were shared by two or more strains. The 3 genotypes from Northern California are closely related as are 3 genotypes from Middle Californian populations which are not nearest neighbors. DNA fingerprinting with the oligonucleotide (GATA)₄ gave compatible results, but the comparison was limited to samples run on one gel. Isoenzyme patterns are compatible with the DNA results, but limited by the very low number of informative polymorphisms. The clustered relationship among genotypes within a species and their geographic distribution suggests very restricted genetic recombination and an origin of new populations from randomly dispersed achenes within the range of the species.     

Comparative analyses of genetic/epigenetic diversities and structures in a wild barley species (Hordeum brevisubulatum) using MSAP, SSAP and AFLP

We analyzed genetic diversity and population genetic structure of four artificial populations of wild barley (Hordeum brevisubulatum); 96 plants collected from the Songnen Prairie in northeastern China were analyzed using amplified fragment length polymorphism (AFLP), specific-sequence amplified polymorphism (SSAP) and methylation-sensitive amplified polymorphism (MSAP) markers. Indices of (epi-)genetic diversity, (epi-)genetic distance, gene flow, genotype frequency, cluster analysis, PCA analysis and AMOVA analysis generated from MSAP, AFLP and SSAP markers had the same trend. We found a high level of correlation in the artificial populations between MSAP, SSAP and AFLP markers by the Mantel test (r > 0.8). This is incongruent with previous findings showing that there is virtually no correlation between DNA methylation polymorphism and classical genetic variation; the high level of genetic polymorphism could be a result of epigenetic regulation. We compared our results with data from natural populations. The population diversity of the artificial populations was lower. However, different from what was found using AFLP and SSAP, based on MSAP results the methylation polymorphism of the artificial populations was not significantly reduced. This leads us to suggest that the DNA methylation pattern change in H. brevisubulatum populations is not only related to DNA sequence variation, but is also regulated by other controlling systems.     

Amplification of DNA from native populations of soil bacteria by using the polymerase chain reaction.

Specific DNA sequences from native bacterial populations present in soil, sediment, and sand samples were amplified by using the polymerase chain reaction with primers for either "universal" eubacterial 16S rRNA genes or mercury resistance (mer) genes. With standard amplification conditions, 1.5-kb rDNA fragments from all 12 samples examined and from as little as 5 micrograms of soil were reproducibly amplified. A 1-kb mer fragment from one soil sample was also amplified. The identity of these amplified fragments was confirmed by DNA-DNA hybridization.     

Amplification of DNA from native populations of soil bacteria by using the polymerase chain reaction.

Specific DNA sequences from native bacterial populations present in soil, sediment, and sand samples were amplified by using the polymerase chain reaction with primers for either "universal" eubacterial 16S rRNA genes or mercury resistance (mer) genes. With standard amplification conditions, 1.5-kb rDNA fragments from all 12 samples examined and from as little as 5 micrograms of soil were reproducibly amplified. A 1-kb mer fragment from one soil sample was also amplified. The identity of these amplified fragments was confirmed by DNA-DNA hybridization.