基于AFLP标记的中国西藏近缘野生大麦遗传多样性分析

选取7对引物组合, 构建了36份西藏近缘野生大麦和4份栽培大麦的AFLP指纹图谱, 共获得清晰可辨的条带227条, 其中多态性带194条, 占85.46％, 从DNA分子水平显示出所试材料遗传多样性较为丰富。计算得各样品间的遗传距离(欧氏距离)介于2.646~10.488之间, 应用离差平方和法对供试材料的AFLP数据结果进行聚类, 建立了40份大麦材料的AFLP树状图, 聚类结果将40份材料分为5类, 进一步揭示出供试材料间遗传背景的相似性和复杂性。结合Nei’s遗传一致性分析结果, 发现近缘六棱野生大麦较之近缘二棱野生与栽培大麦的亲缘关系更近, 支持栽培大麦是从野生二棱大麦起源, 而野生六棱大麦是进化过程中的过渡类型的大麦系统发生观点。     

栽培灯盏花DNA指纹图谱研究及遗传多样性分析

目的:采用ISSR标记构建云南省栽培灯盏花DNA指纹图谱并进行遗传多样性分析.方法:从20对候选引物中筛选出7对引物对云南10个地方栽培灯盏花进行扩增,通过扩增带型的差异构建其DNA指纹图谱,利用Popgen32、NTSYS2软件进行遗传一致性系数和遗传多样分析.结果:7对引物在10份共扩增出48条谱带,其中33条具有多态性,占68.8％,表明各居群的遗传差异较大.10个地方种质资源被聚为2个大类,野生居群被聚为一类,栽培灯盏花聚为一类,其中栽培灯盏花有两大亚类.结论:通过构建DNA指纹图谱容易地把灯盏花种植资源相互区分鉴别出来,10个地方灯盏花种质资源遗传多样性丰富,栽培灯盏花与野生具有差异,栽培种质材料之间也有差异.     

双孢蘑菇遗传多样性分析

应用AFLP指纹技术对双孢蘑菇的20个野生菌株和5个栽培品种的遗传多样性进行了研究。AFLP指纹揭示出20个野生异核体菌株所固的基因型。5个商业品种表现出比较一致的AFLP指纹,但也显示出它们之间的一些差别。由单孢分离获得的同核体菌株携带着部分异核体菌株的AFLP指纹;由同一子实体分离得到的大部分单孢菌株是异核体菌株,它们具有与其亲本一致的AFLP指纹。UPGMA分析揭示出2个与地理分布（美洲、欧洲）和相对应的组。研究结果表明：（1）在野生菌株之间存在着明显的遗传差异;（2）大多数单孢分离的菌株具有与母本一致的遗传物质;（3）野生菌株间的遗传变异大于栽培品种间的变异;（4）在双孢蘑菇的遗传多样性分析中,AFLP技术是非常有效的工具。     

湖北星斗山台湾杉居群的遗传多样性研究

采用随机扩增多态性DNA(RAPD)方法对孑遗植物台湾杉分布于湖北星斗山的野生和栽培居群共42个个体进行了遗传多样性分析。从60个随机引物中筛选出13个引物,共扩增出155条清晰谱带,多态位点百分率为72.9%。野生居群和栽培居群的多态位点百分率分别为65.81%和59.35%;Nei’s基因多样性分别为0.2001和0.1911;Shannon’s信息指数分别为0.3120和0.2946。野生居群的遗传多样性高于栽培居群,表明这些原生古树的后代出现了一定程度的遗传衰退。用UPGMA方法对42个单株聚类可知,来自野生群体的个体和来自栽培群体的个体明显地各聚为一类。尽管孑遗物种台湾杉在湖北仅有一个野生群体,且为零星分布,总个体数40多株,但仍保持较高的遗传多样性水平,这说明遗传多样性不是导致其种群濒危的主要原因,导致台湾杉种群濒危的原因可能与台湾杉自然种群及群落所处生境的直接破坏及其本身生物学和生态学特性所导致的自然更新不良有着密切的联系。     

陆地棉×比克氏棉育成种质系的同工酶和RAPD分析

为了从分子水平上检验野生棉遗传特性向陆地棉转育的结果以及育成种质之间的遗传差异,通过等电聚焦电泳和RAPD技术,对来自科遗181陆地棉品系与野生比克氏棉杂交后代的6个遗传稳定的不同种质系及其亲本的过氧化物酶同工酶图谱和DNA指纹图谱进行了分析。结果如下（1）6个种质系的基本酶谱特征均倾向于陆地棉亲本,但在其中2个种质系的过氧化物酶图谱中,观察到各具有1条等电点值（PI）为4.85的野生亲本的特征带;（2）DNA指纹图谱的分析表明,不同种质系之间在基因组水平上具有高度的异质性。并且在OPO10和OPO112个引物的扩增图谱中观察到4个育成种质系具有野生比克氏棉亲本的特征带。     

杨树重要品种(无性系)的AFLP指纹分析

品种的准确鉴定及其遗传相关性的了解对杨树育种和品种管理具有非常重要的意义。本试验采用AFLP对来自青杨组和黑杨组的21个重要杨树品种（无性系）的鉴定与遗传相关性进行了研究。结果显示,筛选的4对AFLP引物总共产生了181条多态性带,尤其是每对引物对每个品种都产生了独特的指纹图谱;聚类分析和多维尺度分析将试验材料大体上分为五类,结果不仅显示了组间不同品种的差异,而且大体上区分了我国原生品种和外来品种。本研究表明,所有品种都可被筛选的引物准确鉴定,遗传相关性的推断结果与它们的系谱或分类基本一致。另外,本研究还表明AFLP技术完全可用于大规模地构建杨树树种DNA指纹图谱、进行树种鉴定和遗传相关性的研究．     

用AFLP分子标记探讨吴茱萸的遗传多样性

用AFLP标记分析研究吴茱萸与其变种石虎和疏毛吴茱萸之间的遗传多样性.采用AFLP技术,从18对引物中筛选出3对引物.对19株不同地域的3种吴茱萸AFLP指纹图谱进行了分析,计算不同品种间的遗传距离和构建吴茱萸的聚类分析树状图.3对引物共扩增出93条带,其中57条(61.3%)呈多态性,吴茱萸种质内遗传距离为0.059～0.765;在相似系数0.48的水平上.19份吴莱萸材料可以大致分为2个类群.吴茱萸不同品种问遗传距离差异较大,遗传分类在一定程度上与传统的分类方法是一致的,这暗示遗传变异与地域分布有相关性.     

血满草RAPD-PCR反应体系的建立与DNA指纹图谱研究

目的:获得稳定性、重复性好的RAPD-PCR反应体系,并构建血满草DNA指纹图谱和进行遗传多样性分析。方法:以血满草3个居群11个个体的幼叶为材料,CTAB法提取基因组DNA,从模板浓度、引物浓度、d NTPs浓度和Taq DNA聚合酶的用量,构建最佳的RAPD-PCR反应体系,通过扩增带型的差异构建其DNA指纹图谱,利用Popgen32、NTSYS2进行遗传多样性分析。结果:3个RAPD引物扩增血满草3个居群11个样品共获得27条可靠、清晰和重复性高的条带,其中17条是多态条带。引物SBSA5和SBSA11扩增条带组合可以构建11个样品的个体特异DNA指纹图谱。供试血满草居群间遗传距离在0.1493~0.2312之间;居群内的遗传多样性分别为0.1102、0.0153、0.2294。遗传距离和相似性聚类分析表明,样品间的遗传距离与居群的地理分布关系不明显。结论:RAPD分子标记在构建血满草DNA指纹图谱是可行的,由其构建的指纹图谱可以将供试个体相互区分鉴别出来。无论是居群间还是居群内,供试血满草的遗传多样性均较低,居群间的遗传分化较小,可能还在属于同一个大的居群。     

青海湖不同支流中青海湖裸鲤的AFLP遗传多样性分析

为研究青海湖不同支流中青海湖裸鲤(Gymnocypris przewalskii)群体间的遗传差异,我们对采自于不同支流中的群体进行了扩增片段长度多态性分析(AFLP)。应用10对多态性引物在6个青海湖裸鲤群体中共扩增位点348个,其中多态性位点184个,多态性条带占总扩增条带的比例为52.9%;筛选得到79个特异位点,构建了AFLP指纹图谱,根据谱带特征可以将不同支流中的青海湖裸鲤准确区分。遗传聚类分析对比结果表明:当遗传相似系数为0.87时,将六条河流的裸鲤归为为四支,沙柳河、甘子河、黑马河各归一支,哈尔盖河、泉吉河、布哈河为同一支。研究结果为青海湖裸鲤遗传多样性检测和亲本选育提供了技术参数,对青海湖裸鲤资源保护和人工增殖放流工作中野生亲鱼的选择策略具有指导意义。     

东北梅花鹿居群内亲缘关系的AFLP指纹分析

用扩增片段的长度多态性(amplified fragment length polymorphism,AFLP)标记分析研究了东北梅花鹿同一居群内27个个体的亲缘关系,并以此作为优良种鹿选育种的辅助手段。筛选出9对AFLP引物组合,用EcoRⅠ/MseⅠ双酶切,对27只东北梅花鹿基因组DNA进行AFLP检测,共获得15 169条扩增带,检测出多态性条带11 443,多态性比率78.43%,平均每对引物检测到1 271个多态性位点。群体内相似系数AFLP研究结果,平均为0.7841(0.6809-0.8648),27只鹿聚为Ⅰ和Ⅱ两大类群,Ⅱ大类群分为5组,说明该鹿群个体间有较丰富的遗传变异,且与人工定向选配种有关。Ⅱ-4组群体内相似系数最高,在0.82以上,个体之间遗传距离最小在0.1354-0.1563之间,与繁殖记录的亲缘关系基本一致。该研究表明AFLP指纹技术用于梅花鹿遗传多态性分析,品种鉴定及亲缘关系分析是可行的。     

Characteristics and Distributions of Isozymes of Superoxide Dismutase in Leaf Cella of Peanut and wheat

The isozymes of superoxide dismutase (SOD) in leaves of wheat and peanut were investigated and identified by isoelectric focusing gel electrophoresis. The total four bands were found in wheat leaves, among them the band with the highest mobility was proved to be the chloroplastic SOD, the others with intermediate mobilities were probably cytoplasmic SODs. It was observed that there was apparent difference between patterns of SOD isozymes from leaves of wild and cultivated peanut. Leaves of wild peanut contain three different types of SOD isozyme (band A, band B and band C). Band A and B were cyanide-sensitive, hence they should be Cu-Zn-SOD. Band C was cyanide- insensitive and might be Mn-SOD located in mitochondria. The major part of SOD activity in cultivated peanut leaves was concentrated in chloroplasts, and it was cyanide-sensitive. The experiments of SOD activity inhibition by KCN showed that leaves of cultivated peanut contained more Cu-Zn-SOD. On the contrary, the percentage of non-Cu-Zn-SOD was highter in wild species than in cultivated. The PI values of distinct bands were determined and the significance of this study for the further evaluation of resistance of peanut plant is discussed.     

小麦和花生叶细胞内超氧物歧化酶(SOD)同工酶的特性和分布

用等电聚焦凝胶电泳方法对小麦、花生叶内超氧物歧化酶同工酶进行了研究,在栽培小麦叶中发现4条 SOD 同工酶谱带。证明位于图谱下方迁移率最大的强单带是叶绿体 SOD,其它3个可能是细胞质 SOD。对野生花生和栽培花生的比较研究结果表明,前者叶中包含3类不同的谱带(文中 A、B 和 C 带)。A 带及 B 带对氰化物敏感,是 Cu-Zn-SOD,C 带不敏感,可能是线粒体 Mn-SOD。栽培花生叶内 SOD 大部集中在叶绿体中,主要是 Cu-Zn-SOD。本文测定了各谱带的 PI 值,并就两类花生叶中 SOD 的不同特性和分布可能对其生物保护功能的影响作了讨论。     

Genetic structure of the endangered plant Neolitsea sericea (Lauraceae) from the Zhoushan archipelago using RAPD markers

BACKGROUND AND AIMS: The Zhoushan archipelago is the largest archipelago in China. It separated from the mainland about 9000 years ago due to rising sea levels and climate change. Because of the long-term influences of human activities, the original forest vegetation on the large islands has been badly damaged and its plant diversity reduced. METHODS: Levels and patterns of genetic diversity in 114 individuals from six natural populations and four cultivated populations of the insular endangered plant Neolitsea sericea (Lauraceae) on the Zhoushan archipelago were assessed using random amplified polymorphic DNA (RAPD) markers. KEY RESULTS: A total of 99 discernible loci were obtained for all populations using ten primers, 50.5 % of which were polymorphic [percentage of polymorphic bands (PPB)=50.5 %]. Despite being a woody, long-lived, perennial, outcrossing and insect-pollinated plant, N. sericea exhibited low levels of genetic variation. The cultivated populations (PPB=18.9 %, HE=0.060, S=0.092) were genetically less diverse than the natural populations (PPB=23.1 %, HE=0.082, S=0.123). Based on analysis of molecular variance, a high degree of among-population differentiation was revealed for both natural (0.387) and cultivated populations (0.598). CONCLUSIONS: Removal of plants from the wild for horticulture purposes has eroded the level of genetic variation of N. sericea. Low levels of genetic diversity and a high degree of population differentiation indicate that management strategies should include conservation of natural habitats occupied by all six wild populations, and sampling of germplasm resources from multiple seed sources.     

Genetic structure in cultivated and wild carrots (Daucus carota L.) revealed by AFLP analysis

Genetic variation within and among five Danish populations of wild carrot and five cultivated varieties was investigated using amplified fragment length polymorphism (AFLP). Ten AFLP primer combinations produced 116 polymorphic bands. Based on the marker data an UPGMA-cluster analysis and principal component analysis (PCA) separated the Daucus collections into three groups, consisting of the wild populations, the old varieties, and the recently bred varieties. The genetic distance between the wild populations reflected the physical distance between collection sites. Analysis of genetic diversity showed that the old varieties released between 1974 and 1976 were more heterogeneous than the newly developed F₁ hybrid varieties. The analysis of molecular variation (AMOVA) showed that the major part of the genetic variation in the plant material was found within populations/varieties. The presence of markers specific to the cultivated carrot makes it possible to detect introgression from cultivated to wild types. Received: 6 October 1999 / Accepted: 4 November 1999     

Superoxide Dismutase (SOD)Zymogram Patterns and Their Geographical Distribution of Wild (Glycine soja) and Cultivated Soybean (G. max) in China

The purpose of this study was to examine the superoxide dismutase (SOD) zymogram patterns, their frequency and geographical distribution of wild (Glycine soja) and cultivated soybean (G. max) in China. Seeds of 226 wild soybean germplasms and 104 cultivated soybean cultivars (land races) were collected from all provinces and autonomous regions in China except Taiwan, Xinjiang and Qinghai provinces About 50 embryos per wild soybean germplasm and I0 embryos per cultivated soybean cultivars were used for test. Vertical polyacrylamide gel electrophoresis and a stainning system modified after Luo (1984)were used. The Japanese GS- 930 Scanner was used in gel-plate scanning. In program scanning the maximum and minimum absorption wavelength were 700 and 550 nm respectively. The results showed that: 1. Six zymogram patterns were found in soybean (Fig. 1, 2). Wild soybean displayed five patterns (Ⅰ, Ⅱ, Ⅳ Ⅴ, Ⅵ), while the cultivated soybean displayed only two patterns (Ⅱ, Ⅲ). 2. Fourty six percent of wild germplasms gave an 7-band zymogram (Table Ⅰ) (pattern Ⅰ), fourty nine percent had a 6th and 7th band with faster mobility (pattern Ⅱ), about two percent produced a 6-band zymogram which lacked the SODc4 band (pattern Ⅳ), about two percent had a 5-band pattern which lacked the SODc,c4 bands (pattern Ⅴ), and only one germptasm displayed a 5-band zymogram which lacked SODb2b3 bands (pattern Ⅵ). 3. More than ninty eight percent of cultivated cultivars belonged to pattern Ⅱ, only about two percent belonged to pattern Ⅲ. 4. The geographical distribution of frequency of pattern Ⅱ between wild and cultivated soybean was most close in 36–51º N area. The difference of zymograms between G. soja and G. max, and the problems of the origional area and evolution of soybean were discussed.     

The genetic diversity of the Vigna angularis complex in Asia.

A selected set of accessions of components of the azuki bean (Vigna angularis) complex comprising 123 cultivated accessions and 23 wild or weedy accessions from Bhutan, China (including Taiwan), India, Japan, Korea, and Nepal was analyzed using amplified fragment length polymorphism (AFLP) methodology. Using 12 AFLP primer pairs, 580 unambiguous bands were generated, 313 (53.9%) of which were polymorphic among azuki bean accessions. All 580 bands were used to assess phenotypic (band) and genetic (nucleotide) diversity among the 146 azuki bean accessions. The results indicate five major groups of azuki bean germplasm primarily associated with geographic origin of accessions and their status: wild, weedy, or cultivated. These five groups are (i) Himalayan wild, (ii) Nepal-Bhutan cultivated, (iii) Chinese wild, (iv) Taiwan wild - Bhutan cultivated, and (v) northeast Asian accessions. Within the northeast Asian accessions, three subgroups are present. These consist of (v1) Japanese complex - Korean cultivated, (v2) Japanese cultivated, and (v3) Chinese cultivated accessions. The results suggest domestication of azuki bean occurred at least twice, once in the Himalayan region of southern Asia and once in northeast Asia. The remarkable diversity of azuki bean germplasm in the Himalayan region compared with other regions suggests this is a rich source of germplasm for plant breeding. The results suggest there are important gaps in the germplasm collections of azuki bean and its close relatives from various parts of Asia and that specific collecting missions for Vigna germplasm related to azuki bean in the highlands of subtropical Asia are needed.     

Patterns of Genetic Variation in in and ex situ Populations of the Threatened Chilean VineBerberidopsis corallina , Detected Using RAPD Markers

Berberidopsis corallina Hook. f. (Berberidopsidaceae) is a threatenedvine, endemic to the temperate rainforests of southern Chile.A RAPD analysis was carried out to assess the extent of geneticvariation in remaining wild populations and in British cultivatedplants, to assess the value of the latter for ex situ conservation.A total of 90 individuals (54 wild, 35 cultivated) were analysed,and a total of 54 polymorphic bands produced. A pairwise distancemeasure calculated from the RAPD data was used as input forprincipal coordinate analysis (PCO) and analysis of molecularvariance (AMOVA). AMOVA indicated that an exceptionally highproportion of total genetic variation (54.8%) was recorded amongpopulations; pairwise     

Genetic characterization and phytochemical analysis of wild and cultivated populations of Scutellaria baicalensis

Scutellaria baicalensis was collected from four wild and four cultivated populations from different locations in China. Forty-two samples were analyzed using random amplification of polymorphic DNA (RAPD) techniques for genetic profiling, and high performance liquid chromatography (HPLC) techniques to determine the flavonoid content. The selected 23 RAPD primers yielded a total of 838 clear and reproducible bands of which 237 were found to be polymorphic. The wild population exhibited higher polymorphism than that of the cultivated population. The dendrogram generated by the UPGMA method via Nei's genetic distance revealed three distinct genotypes from the cultivated populations and several branches from the wild populations. The contents of baicalin and wogonoside in dried roots of the samples ranged from (w/w) 8.63 to 17.84%, and from 1.99 to 4.21%, respectively, whereas their aglycones, baicalein and wogonin, were within the range of only 0.04-0.23%. The total content of the four flavonoids varied from 9.45 to 26.24%. Comparatively, the cultivated populations contained much higher levels of baicalin and total flavonoids than those in the wild populations. The results from genetic characterization and phytochemical analysis demonstrated that the quality variation of this drug was mainly determined by extrinsic environmental or agricultural factors, rather than by genetic differences. Our findings can be used for the commercial production and germplasm management of this medicinal plant.     

用RAPD和ISSR检测的橡胶树野生种质和栽培品种的遗传多样性

用19个RAPD引物和12个ISSR引物对14份野牛橡胶树种质和我国的37份栽培品种进行了遗传多样性分析。RAPD引物共产生132条带,多态性带占88．6％,相似系数变化范围在0．432—0．947。ISSR引物其产生101条带,多态性带占87．1％,相似系数为0．505—0．941。平均基因杂合度分析表明野生种质比栽培品种具有较高的遗传多样性。根据UPGMA法对51份材料进行聚类分析,结果表明,ISSR分析中所有材料可分为2类：第一类为野生种质,第二类为栽培品种：而RAPD分析中野牛种质和栽培品种不能被分为明显的两人类。虽然ISSR和RAPD的聚类分析结果存在差异,但对两种方法进行的相关分析表明,他们之间仍存在极显著相关性,相关系数为0．574。品种PR107、热研217等一些栽培品种可以通过特异带在51份供试材料中被区分开。这些结果可以对橡胶树的育种上作起到一定的指导作用,同时RAPD和ISSR技术也是进行橡胶树品种鉴定和遗传多样性研究的有效手段。     

产杜仲黄酮内生真菌的初步研究

由杜仲植株的根、茎、叶分离到内生真菌,分别对各菌株进行液体培养,筛选能产生与杜仲药材相同或相似黄酮类化合物的内生真菌。将培养物适当处理后,根据黄酮类化合物所特有的颜色反应进行初筛,然后根据TLC分析和紫外-可见光分光光度法测定结果进行复筛,结果从杜仲植株共分离得到44株内生真菌,其中1株(DZY5)能产出与其宿主相同或相似的黄酮类物质。