PIXE and ICP-AES analysis of early glass unearthed from Xinjiang (China)

Early glasses (about 1066 BC-220 AD) unearthed from Xinjiang of China were chemically characterized by using PIXE and ICP-AES. It was found that these glasses were basically attributed to PbO-BaO-SiO2 system, K2O-SiO2 system, Na2O-CaO-SiO2 system and Na2O-CaO-PbO-SiO2 system. The results from the cluster analysis showed that some glasses had basically similar recipe and technology. The PbO-BaO-SiO2 glass and the K2O-SiO2 glass were thought to come from the central area and the south of ancient China, respectively. The part of the Na2O-CaO-SiO2 glass (including the Na2O-CaO-PbO-SiO2 glass) might be imported from Mesopotamia, while the other part might be locally produced. (c) 2005 Elsevier B.V. All rights reserved.

Genetic structure analysis of natural Sargassum muticum (Fucales, Phaeophyta) populations using RAPD and ISSR markers

Sargassum muticum is important in maintaining the structure and function of littoral ecosystems, and is used in aquaculture and alginate production, however, little is known about its population genetic attributes. In this study, random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers were used to investigate the genetic structure of four populations of S. muticum and one outgroup of S. fusiforme (Harv.) Setchell from Shandong peninsula of China. The selected 24 RAPD primers and 19 ISSR primers amplified 164 loci and 122 loci, respectively. Estimates of genetic diversity with different indicators (P%, percentage of polymorphic loci; H, the expected heterozygosity; I, Shannon's information index) revealed low or moderate level of genetic variations within each S. muticum population, and a high level of genetic differentiations were determined with pairwise unbiased genetic distance (D) and fixation index (F-ST ) among the populations. The Mantel test showed that two types of matrices of D and F-ST were highly correlated whether from RAPD (r = 0.9706, P = 0.009) or ISSR data (r = 0.9161, P = 0.009). Analysis of molecular variance (AMOVA) was conducted to apportion the variations among and within the S. muticum populations. It indicated that variations among populations were higher than those within populations, being 55.82% verse 44.18% by RAPD and 55.21% verse 44.79% by ISSR, respectively. Furthermore, the Mantel test suggested that genetic differentiations among populations were related to the geographical distances (r > 0.6), namely, conformed to the IBD (isolation by distance) model, as expected from UPGMA (unweighted pair group method with arithmetic averages) cluster analysis. On the whole, the high genetic structuring among the four S. muticum populations along the distant locations was clearly indicated in RAPD and ISSR analyses (r > 0.9, P < 0.05) in our study.

野大豆和栽培大豆抗盐分子标记的研究

野大豆群体3和群体4属盐渍群体，其个体有的是抗盐的，有的是敏感的，有的是中等抗盐的，本文通过随机扩增多态性DNA (RAPD)和DNA扩增指纹(DAF)分析野大豆群体抗盐性与分子标记之间的关系，从而更好地研究野大豆群体的盐适应机理。通过12个RAPD引物和3个DAF引物扩增发现：引物OPF05，OPF19和OPH02的扩增产物中有与抗盐性可能相关的特异标记，分别是OPF05_(213);OPF19_(4361);OPF19_(1727);OPF19_(1400);OPF19_(700);OPH02_(1350)。这些特异标记在所研究的抗盐植株中都存在;在敏感型植株中都不存在;在中等抗盐植株中有的存在，有的不存在。以上表明野大豆群体的抗盐性与RAPD分子标记有一定的相关性。为进一步研究抗盐性的特异标记，本文对栽培大豆抗盐品种Morgan和文丰七号的特异DAF标记片断8-27_(240) (Zhong et al., 1997)进行了克隆测序，测序结果通过BLASTn程序与基因库中的基因序列进行同源比较，发现上的DNA序列中的19组（每组大约二十到三十个碱基）序列与基因库中的其它基因相应序列有很高的同源性，几乎全部100%同源。尤其目的序列第15个碱基到第33个碱基（共19个碱基）之间的序列与基因库中的25个基因的相应序列的同源性全部是100%，并且与之相比的基因大多来自动物和人。因而推测其有可能是保守区，而不是编码区。进一步用DNASIS软件分析其碱基组成(A/T含量是64.9%，G/C含量是35.1%)并进行翻译，结果同样表明此序列可能是一调控序列并非编码区。至于这段序列是否与抗盐紧密相关，这有待于以后把此序列转到敏感型植株中然后检测其抗性来验证。 本文还通过RAPD分析野大豆群体3和群体4的多态性，发现群体3的多态性明显高于群体4。野大豆群体3的抗盐性大于群体4早已通过生理指标的鉴定，至于多态性与抗盐性之间是否有必然联系，还需进一步研究讨论。利用RAPD数据，通过MEGA软件中的NJ计算遗传距离的方法对群体3和4进行聚类分析，研究野大豆群体间及群体内个体间的亲缘及进化关系，探讨野大豆群体盐适应机理的分子起源。

构树种质资源遗传多样性SRAP分子标记研究

构树（Broussonetia papyrifera L.）为桑科（Moraceae）构树属（Broussonetia）落叶乔木，广泛分布于亚洲东部及太平洋岛屿。它是我国重要的经济林木，具有重要的经济价值，其树皮纤维品质优良，自古就是造纸的优良原料；叶片可用作饲料；果实具有重要的药用价值；环境适应性强，是迅速绿化荒山、荒滩和盐碱地的理想树种。因此对构树这些特性的深入研究和开发利用具有非常重要的实际应用价值。本研究以日本和国内构树主要分布区域的的10种生态型及杂交构树共23份材料，摸索并改进了构树DNA的提取方法，建立了稳定的SRAP分子标记体系，以杂交构树组培苗为材料从120对引物组合中筛选出条带信息较高的17对SRAP引物，以这些引物对23份构树材料进行PCR扩增和标记分析。 本研究取得的主要结果如下： （1）本实验首次将SRAP技术应用于构树的研究中，建立起构树稳定的SRAP-PCR反应体系；实验中对影响扩增的5个主要因素进行了优化，确定20 μL PCR反应体系中各因素最适浓度：模板DNA浓度80 ng/(20 μL)，Mg2+浓度2.0 mM，dNTP浓度0.6 mM，引物浓度0.8 mM，Taq 酶浓度1.5 U/(20 μL)。 （2）从120对引物组合中筛选出来的17对SRAP引物对21份不同生态型构树样本（21份材料指的是除两份杂交构树材料外的其它生态型构树，以下同）进行PCR扩增，共扩增出439条带，平均每对引物25.5条，其大小介于100～1,000 bp之间，其中多态性条带319条，占总数的72.67%。 （3）用Popgene1.32软件进行分析，计算出Shannon信息指数（I）值为0.2275（0.2042），物种水平的Nei基因多样性（H）值为0.1336（0.1436），表明各生态型构树之间的平均遗传多态性不高，中国大陆各生态型构树Shannon信息指数（I）值仅为0.1675（0.2271），物种水平的Nei基因多样性（H）值为0.1039（0.1540），群体遗传多样性较低，构树的遗传分化主要存在于中国和日本之间。 （4）用NTSYS-2.10e软件进行聚类分析，发现不同生态型构树按距离关系远近及分布区域可划分为不同类群。在遗传相似系数0.57附近，对21份材料进行聚类分析发现其可分为两个群体，一类为日本生态型，另一类为中国生态型，表明日本构树与中国野生种构树种源遗传相似性较小。中国各生态型构树在遗传相似系数0.91处可分为5类，总体而言，中国各地区之间的构树遗传相似度较高。对杂交构树分析表明，其亲缘关系与日本构树（母本）更接近。 （5）日本及杂交构树的SCAR标记。本研究找到两条日本及杂交构树的特异性条带，回收、测序，再根据序列往里重新设计引物，转变成稳定性更好，更直观的SCAR标记，这为挑选性状优良的日本及杂交构树提供重要的参考，对其育种有一定的指导意义。其中一条片段经与NCBI数据库比对发现与拟南芥磺基转移酶家族基因具有较高的同源性。

紫薇种质资源的研究

本文回顾了紫薇在国内外的栽培历史。在中国，紫薇的栽培虽历史悠久，但种质资源缺少系统的研究。本研究通过宏观与微观的技术和方法，探索紫薇种间及品种间的演化关系，并为今后开展种质鉴定、 品种分类以及有计划、有目的的进行新品种的培育提供依据，以期达到紫薇种质资源多样性保护的目的。主要的结果及结论： 1．认为紫薇栽培的历史是紫薇不同种质进行组合和渗透的历史，也在一定程度上反映了品种演化的历史。并提出除了抗病、矮型紫薇的育种外，还要重视重瓣紫薇的育种。 2．通过对紫薇的形态学性状进行聚类分析，寻找到了评价紫薇种质资源的标准，进而提出栽培观赏植物种质资源的评价标准， 即将其性状分为种源性状和观赏性状，这种对观赏植物性状的分解反映了栽培植物不同于野生植物的特点。同时，对其形态学性状进行主分量分析表明，花色与其它营养器官性状相关性不大，很难从其营养器官性状推测花的颜色。 3．通过对紫薇的分子生物学研究，建立了紫薇的DNA提取流程及RAPD扩增体系，利用RAPD标记可以进行紫薇种质的鉴定及探求不同种质间的关系。 4．综合形态学及分子生物学的结论，建立了紫薇种质资源的划分体系，在此基础上，建成了种质资源圃及紫薇种质资源数据库。

云南自然保护区珍稀濒危哺乳动物分布格局的聚类分析

　采用聚类分析测定云南地区9 个国家自然保护区珍稀濒危哺乳动物分布格局的相似性。以编码为 1 的有物种分布及编码为0 的无物种分布作为二元状态,建立二元数据列联表,应用类与类的空间距离Dpq聚类。 结果表明,珍稀濒危哺乳动物的分布未出现纬度梯度变异,横断山区省为一类,西南区省和华南区省与云南动物地 理区划系统基本一致,相邻保护区无量山与哀牢山分别为一类。在保护区之间应建立生物廊道,以促进物种基因 的交流,扩大面积较小的保护区,合并面积相对小的保护区。

Relationships between Testate Amoebae Assemblages (Protozoa) and Geographic Factors in Yunnan Plateau Lakes, China

Knowledge of the effect of geographic factors on the assemblages of protozoan testate amoebae is still limited, despite there having been a number of studies on this fauna. We applied statistical analyses to data on the distribution of testate amoebae from nine major lakes in the Yunnan Plateau, southwest China. Cluster analysis, based on community structure, separated the lakes into two groups - the oligotrophic/mesotrophic lakes and the hypercutrophic lakes - confirming the idea that the testate amoebae assemblages in lakes are closely related to the trophic status. Additionally, within the oligotrophic/mesotrophic lakes, there was distinct geographic clustering. Linear regression analysis and the Mantel test both revealed that similarity of species composition decreased with increasing geographic distance among the oligotrophic/mesotrophic lakes.

Morphological variability between wild populations and inbred stocks of a Chinese minnow, Gobiocypris rarus

Gobiocypris rarus, a small, native cyprinid fish, is currently widely used in research on fish pathology, genetics, toxicology, embryology, and physiology in China. To develop this species as a model laboratory animal, inbred strains have been successfully created. In this study, to explore a method to discriminate inbred strains and evaluate inbreeding effects, morphological variation among three wild populations and three inbred stocks of G. rarus was investigated by the multivariate analysis of eight meristic and 30 morphometric characters. Tiny intraspecific variations in meristic characters were found, but these were not effective for population distinction. Stepwise discriminant analysis and cluster analysis of conventional measures and truss network data showed considerabe divergence among populations, especially between wild populations and inbred stocks. The average discriminant accuracy for all populations was 82.1% based on conventional measures and 86.4% based on truss data, whereas the discriminant accuracy for inbred strains was much higher. These results suggested that multivariate analyses of morphometric characters are an effective method for discriminating inbred strains of G. rarus. Morphological differences between wild populations and inbred strains appear to result from both genetic differences and environmental factors. Thirteen characters, extracted from stepwise discriminant analysis, played important roles in morphological differentiation. These characters were mainly measures related to body depth and head size.

Studies on the restoration succession of PFU microbial communities in a pilot-scale microcosm

In order to imitate the restoration succession process of natural water ecosystem, a laboratory microcosm system of constant-flow-restoration was designed and established. A eutrophycation lake, Lake Donghu, was selected as the subject investigated. Six sampling stations were set on the lake, among which the water of station IV was natural clean water, and others were polluted with different degrees. Polyurethane foam unit microbial communities, which had colonized in the stations for a month, were collected from these stations and placed in their respective microcosms, using clean water of station IV to gradually replace the water of these microcosms. In this process, the healthy community in clean water continuously replaced the damaged communities in polluted water, the restoration succession of the damaged communities was characterized by weekly determination of several functional and structural community parameters, including species number (S), diversity index (DI), community pollution value (CPV), heterotrophy index (HI), and similarity coefficient. Cluster analysis based on similarity coefficient was used to compare the succession discrepancies of these microbial communities from different stations. The ecological succession of microbial communities during restoration was investigated by the variable patterns of these parameters, and based on which, the restoration standards of these polluted stations were suggested in an ecological sense. That was, while being restored, the water of station 0 (supereutrophycation) should be substituted with natural clean water by 95%; station I (eutrophycation), more than 90%; station II (eutrophycation), more than 85%; station III (eutrophycation), about 85%; station V (mesoetitrophycation), less than 50%. The effects of the structural and functional parameters in monitoring and assessing ecological restoration are analyzed and compared. (C) 2007 Elsevier Ltd. All rights reserved.

Development of the microbial communities in Lake Donghu in relation to water quality

There is increasing recognition that protozoa is very useful in monitoring and evaluating water ecological healthy and quality. In order to study the relationship between structure and function of protozoan communities and water qualities, six sampling stations were set on Lake Donghu, a hypereutrophic subtropical Chinese lake. Microbial communities and protists sampling from the six stations was conducted by PFU (Polyurethane foam unit) method. Species number (S), diversity index (DI), percentage of phytomastigophra, community pollution value (CPV), community similarity and heterophy index (HI) were mensurated. The measured indicators of water quality included total phosphorus (TP), dissolved oxygen (DO), Chemical oxygen demand (COD), NH4 (+), NO2 (-) and NO3-. Every month water samples from stations I, II, III, IV were chemically analyzed for a whole year, Among the chemically analyzed stations, station I was the most heavily polluted, station II was the next, stations III and IV had similar pollution degrees. The variable tendencies of COD, TP, NH3, NO2-, NO3-, and DO during the year was approximately coincident among the six stations. Analysis from the community parameters showed that the pollution of station 0 was much more serious than others, and station V was the most slight. Of the community parameters, CPV and HI were sensitive in reflecting the variables of the water quality. Community similarity index was also sensitive in dividing water qualities and the water quality status of different stations could be correctly classified by the cluster analysis. DI could reflect the tendency of water quality gradient, species number and percentage of Phytomastigophora was not obvious in indicating the water quality gradient.

Population genetic structure of Carchesium polypinum (Ciliophora : Peritrichia) in four chinese lakes inferred from ISSR fingerprinting: High diversity but low differentiation

Although the peritrichous ciliate Carchesium polypinum is common in freshwater, its population genetic structure is largely unknown. We used inter-simple sequence repeat (ISSR) fingerprinting to analyze the genetic structure of 48 different isolates of the species from four lakes in Wuhan, central China. Using eight polymorphic primers, 81 discernible DNA fragments were detected, among which 76 (93.83%) were polymorphic, indicating high genetic diversity at the isolate level. Further, Nei's gene diversity (h) and Shannon's Information index (I) between the different isolates both revealed a remarkable genetic diversity, higher than previously indicated by their morphology. At the same time, substantial gene flow was found. So the main factors responsible for the high level of diversity within populations are probably due to conjugation (sexual reproduction) and wide distribution of swarmers. Analysis of molecular variance (AMOVA) showed that there was low genetic differentiation among the four populations probably due to common ancestry and flooding events. The cluster analysis and principal component analysis (PCA) suggested that genotypes isolated from the same lake displayed a higher genetic similarity than those from different lakes. Both analyses separated C. polypinum isolates into subgroups according to the geographical locations. However, there is only a weak positive correlation between the genetic distance and geographical distance, suggesting a minor effect of geographical distance on the distribution of genetic diversity between populations of C. polypinum at the local level. In conclusion, our studies clearly demonstrated that a single morphospecies may harbor high levels of genetic diversity, and that the degree of resolution offered by morphology as a marker for measuring distribution patterns of genetically distinct entities is too low.

DNA polymorphism of the plankton community and its relationships to species composition in the Three Gorges Reservoir Region of the Yangtze River

We examined DNA polymorphism of the plankton community in the Three Gorges Reservoir Region of Yangtze River and studied its relationships to species composition. Samples of the plankton community were collected from nine sampling sites and analyzed by random amplified polymorphic DNA (RAPD). Nine of 60 screened primers generated a total of 88 observable 180 to 1400 by bands, all of which were polymorphic. Cluster analysis of the resulting binary format from DNA banding patterns grouped the target communities into three clusters. The topology of the constructed diagram from species composition data was generally similar to that based on RAPD markers.

Microcalorimetry as a possible tool for phylogenetic studies of Tetrahymena

Using isothermal microcalorimetry, the growth power-time curves of three strains of Tetrahymena were determined at 28 degrees C. Their Euclidean distances and cluster analysis diagram were obtained by using two thermokinetic parameters (r and Q(log)), which showed that T. thermophila BF1 and T. thermophila BF5 had a closer relationship. Compared with the single molecular biomarker (ITS1) method, microcalorimetry wasmaybe a simpler, more sensitive andmore economic technique in the phylogenetic studies of Tetrahymena species.