新疆产甘草6个不同地理群体遗传关系的RAPD分析

目的探讨新疆产甘草不同地理群体的遗传多样性。方法用人工栽培品种作为对照 ,采用 2 0个 10碱基随机引物对新疆产甘草 (GlycyrrhizauralensisFisch .exDC) 6个不同地理群体的基因组DNA进行RAPD分析。结果共扩增出 175个位点 ,其中多态性位点 10 2条 ,占 5 8.2 8%。通过计算遗传相似性系数 ,建立了UPGMA聚类图。结论不同地理群体中存在一定的遗传多样性 ;产地相距越远 ,群体间相似性程度越低 ;人工栽培种与同一产地野生种具有相似的遗传特性     

RAPD分子标记对药用植物大花红景天濒危原因分析

目的:利用RAPD分子标记技术研究大花红景天的遗传多样性,分析其濒危的可能原因.方法:利用改良的CTAB法提取各产地大花红景天的总DNA,以40条随机引物随机扩增,利用RAPDdistance1.04软件计算多态位点百分率以及遗传距离,并进行聚类分析.结果:大花红景天表现出较为丰富的遗传多样性,加条随机引物筛选出能扩增出清晰稳定条带的12条随机引物.12条随机引物共扩增出99条DNA条带,其中79条表现出多态性,占总带数的79.8%.各产地大花红景天的遗传距离在0.4784-0.7774之间.通过聚类分析可将5个产地的大花红景天分为2个类群.结论:多态位点百分率表明大花红景天具有丰富的遗传多样性,其濒危的可能原因是由于人类无规则且盲目的采挖造成的.     

不同产地白及遗传多态性的RAPD分析

目的 研究国内不同产地白及的遗传多样性和亲缘关系。方法 利用随机扩增多态DNA （random amplified polymorphic DNA,RAPD）引物,RAPD分子标记技术分析国内50个不同产地的白及。结果 RAPD技术扩增产物经琼脂凝胶电泳检测,从50条引物中筛选出10条（S8,S9,S14,S19,S23,S25,S28,S29,S30,S31）条带清晰的引物,10条引物共扩增出88条DNA条带,其中多态性的DNA条带数目为81条,占总数的92.05%。每个引物能扩增出5~11条DNA条带,平均可扩增出8.8条;扩增最少的引物为S19,扩增出5条DNA条带;扩增最多的引物为S29,扩增出11条DNA条带。而每个引物能扩增的多态性DNA条带数为3~11条,平均可扩增出8.6条。结论 不同产地的白及具有较丰富的遗传多样性,RAPD可有效应用于白及的遗传多样性研究。     

广西产鸡血藤遗传多样性的RAPD与ISSR标记方法的比较研究

目的:比较随机扩增多态性DNA分子标记技术(RAPD)和简单重复序列间区扩增技术(ISSR)两种标记方法,研究广西不同居群鸡血藤遗传多样性的优劣。方法:分别采用RAPD和ISSR标记方法,利用POPGENE 32软件、Ntsys软件、SPSS 17.0软件,对广西不同采集地的9份鸡血藤的遗传多样性进行研究。结果:筛选出的3条RAPD引物及4条ISSR引物扩增后,共得到198和315个位点,多态性位点37和80个,多态性位点比率为18.7%和25.4%,有效等位基因数为1.416 8、1.584 0,基因多样性指数为0.269 4、0.351 3,Shannon多样性指数为0.431 6、0.529 9。ISSR标记均高于RAPD标记,RAPD和ISSR的平均遗传相似系数为0.757 64、0.683 80,表明ISSR检测遗传多样性更灵敏。二者的聚类结果相近,相关系数r为0.847,说明其在0.001水平呈极显著正相关。结论:ISSR标记方法比RAPD标记反映出更多的遗传多样性信息,更适合于广西产不同居群鸡血藤的遗传多样性研究。     

青阳参及其近缘种的RAPD研究

目的 针对青阳参Cynanchum otophyllum Schneid.存在混淆种类和种内变异较大的情况,对青阳参和与其相混淆的隔山消C．wilfordii (Maxim.) Hemsl.和昆明杯冠藤C. wallichii Wight进行种间鉴别,并对青阳参的4个居群进行遗传多样性分析。方法 运用随机扩增多态性DNA(RAPD)技术进行分析,用Shannon指数和Nei指数评价青阳参的遗传多样性,用UPGMA类平均法进行聚类分析亲缘关系。结果 利用筛选出的10个随机引物对供试材料DNA进行随机扩增,共得到72条带,其中多态性带有70条,多态性百分率为97.22%。由Shannon指数计算青阳参4个居群内的遗传多样性比率为68.52％,高于居群间的31.48%;同时由Nei指数估计青阳参4个居群中有69.44％的遗传变异来自居群内,高于居群间的30.56％。采用UPGMA聚类法得出反映各种间亲缘关系的树状图。结论 运用RAPD方法可以进行青阳参和隔山消两个种同昆明杯冠藤的种间鉴别;虽然居群间已有较高分化,但青阳参遗传变异主要分布在居群内。     

青阳参及其近缘种的RAPD研究

目的针对青阳参Cynanchum otophyllum Schneid．存在混淆种类和种内变异较大的情况,对青阳参和与其相混淆的隔山消C．wilfordii（Maxim．）Hemsl．和昆明杯冠藤C.wallichii Wight进行种间鉴别,并对青阳参的4个居群进行遗传多样性分析。方法运用随机扩增多态性DNA（RAPD）技术进行分析,用Shannon指数和Nei指数评价青阳参的遗传多样性,用UPGMA类平均法进行聚类分析亲缘关系。结果利用筛选出的10个随机引物对供试材料DNA进行随机扩增,共得到72条带,其中多态性带有70条,多态性百分率为97．22％。由Shannon指数计算青阳参4个居群内的遗传多样性比率为68．52％,高于居群间的31．48％;同时由Nei4指数估计青阳参4个居群中有69．44％的遗传变异来自居群内,高于居群间的30．56％。采用UPGMA聚类法得出反映各种间亲缘关系的树状图。结论运用RAPD方法可以进行青阳参和隔山消两个种同昆明杯冠藤的种间鉴别：虽然居群间已有较高分化,但青阳参遗传变异主要分布在居群内。     

铁皮石斛野生居群遗传多样性的RAPD分析与鉴别

目的采用RAPD分子标记技术，对铁皮石斛8个野生居群的遗传多样性、亲缘关系以及分子鉴别等进行研究。方法筛选随机引物进行RAPD分析，通过UPGMA聚类，研究铁皮石斛各居群间的遗传关系，构建居群亲缘关系的分子系统树；利用特异性条带对铁皮石斛野生居群进行指纹分析鉴别。结果共筛选出10个有效引物，在8个野生居群材料的RAPD扩增中共得到439个位点，平均每个引物扩增出43.9个位点，每个居群扩增出54.9个位点；在所获得的104条可重现谱带中，9条是单态的，95条是多态的，多态性程度达91.35%，遗传距离在0.590～0.727之间，平均为0.686。结论铁皮石斛居群间遗传差异明显，具有较丰富的遗传多样性；RAPD可以作为铁皮石斛野生居群遗传多态性、居群亲缘关系和分子鉴别研究的有效手段；引物S412可以有效鉴别铁皮石斛的8个野生居群。     

RAPD标记在药用菊花种质鉴定中的应用

目的：探讨药用菊花类型间的遗传关系,为药用菊花资源合理保护利用和新品种选育提供理论依据,也为药用菊花DNA指纹图谱的构建奠定基础。方法：利用改良SDS方法提取药用菊花的基因组DNA,以RAPD分子标记技术,对6种药用菊花进行了遗传多样性和亲缘关系分析。结果：从8个随机引物中筛选出多态性频率高的2个引物。2个引物共扩增出17个DNA条带,其中多态性条带16个,多态性达94%,显示菊花品种内丰富的遗传多样性。扩增的多态性片段在300~1300bp之间。结论：聚类分析结果表明,利用RAPD标记可将全部供试材料区分开,得出了药用菊花种质资源在分子水平上确实存在较大差异。     

不同产地红曲基原菌的RAPD分析

目的：建立中药红曲基原菌的RAPD分析方法,方法：CTAB法提取不同产地红曲基真菌的基因组DNA,琼脂糖凝胶电泳法检测PCR扩增产物。PHYLIP3．5c进行聚类分析,结果：S282等10个随机引物的扩增产物谱带多,特征好,产地间指纹图谱呈现出明显的DNA扩增产物多态性,结论：RAPD分析技术可作为红曲鉴定的有用手段。     

新疆雪莲花6个不同地理群体遗传关系的RAPD分析

目的建立新疆6个不同地理群体的新疆雪莲花(Saussurea involucrata Kar. et Kin.)指纹图谱,进行遗传多样性分析.方法通过20个10碱基随机引物对新疆6个不同地理群体的新疆雪莲花进行聚合酶链反应(PCR)扩增,通过计算遗传相似性系数,建立平均连接聚类法(UPGMA)聚类图.结果共扩增出103个多态位点,建立了它们的基因组DNA指纹图谱.结论不同地理群体中存在丰富的遗传多样性;相距越远,群体间相似性程度越低.     

Genetic ecotoxicology II: population genetic structure in mosquitofish exposed in situ to radionuclides

In 1977, approximately 250 mosquitofish (Gambusia affinis) from a relatively uncontaminated site (Crystal Springs) were transplanted into a small pond on the Department of Energy Oak Ridge Reservation which is heavily contaminated with radionuclides (Pond 3513). Starting in 1992, DNA polymorphism was evaluated using the RAPD (Randomly Amplified Polymorphic DNA) and allozyme genotype techniques to determine if genetic differentiation had occurred between the two populations. Fish from a second radionuclide-contaminated population (White Oak Lake) and another unrelated non-contaminated population (Wolf Creek) were also examined. For the RAPD analyes, 15 RAPD primers (from a total of 40) were found to produce polymorphic banding patterns in at least two of the four populations and subsequently were used to produce a total of 142 bands. Data generated by these RAPD primers indicated an increased genetic diversity in radionuclide-contaminated sites relative to reference sites. Furthermore, the patterns from six RAPD primers produced a higher average number of bands when using DNA from radionuclide- contaminated populations than from non-contaminated, and for three RAPD primers the average number of bands from radionuclide- contaminated populations was lower. In addition, 17 bands occurred at a higher frequency in the radionuclide-contaminated compared to the non-contaminated populations. For the allozyme analyses, it was found that there was a higher percentage of polymorphism and heterozygosity in the radionuclide-contaminated relative to non-contaminated sites. These findings contribute to our understanding of the evolutionary effects of contaminant exposure as well as to the development of population-level biomarkers     

铁皮石斛野生居群基于RAMP标记的遗传多样性评价

本文运用RAMP标记16对引物组合,对铁皮石斛9个野生居群的112个样本进行了检测,共得到123条扩增带,其中86条(占69.92%)具有多态性,每对引物组合可扩增出3～8条多态性带,平均5条,表明铁皮石斛不同居群间存在较丰富的遗传多样性;铁皮石斛居群间遗传相似系数的变化范围为0.250～0.813,平均值为0.550。利用RAMP扩增条带数据进行聚类分析,可将9个铁皮石斛野生居群划分为3个类群,聚类结果表现出较好的地域相关性,显示RAMP标记可以有效地评价铁皮石斛野生居群的遗传多样性及遗传结构。     

Utilization of RAPD markers to assess genetic diversity of wild populations of North American ginseng (Panax quinquefolium)

The Catskill Mountains of New York State are an important source of wild-collected American ginseng (Panax quinquefolium) and, increasingly, of woods-cultivated ginseng. The objective of this study was to assess genetic diversity among 9 different wild ginseng populations in and adjacent to the Catskill Mountain region of New York State and to compare these to wild populations from other states including Kentucky, Tennessee, North Carolina, Pennsylvania, and Virginia, and one cultivated population from Wisconsin. Randomly amplified polymorphic DNA (RAPD) markers were used to estimate the genetic distance among samples from the 15 populations. Pooled DNA from 10 plants of each of 8 New York populations was initially screened with 64 random primers; subsequently, the 15 primers that exhibited the greatest number of reproducible polymorphic markers were selected for further experimentation. Gel electrophoresis with the selected 15 primers produced 124 highly reproducible polymorphic bands. The ratio of discordant bands to total bands scored was used to estimate the genetic distance within and among populations. Multidimensional scaling (MDS) of the relation matrix showed distinctly separate clusters between New York and non-New York populations, indicating separation between these two groupings. The MDS analysis was confirmed using pooled chi-square tests for fragment homogeneity. This study shows that RAPD markers can be used as population-specific markers for Panax quinquefolium, and may eventually be utilized as markers for ginsenoside assessment.     

鱼腥草种质资源的RAPD分析

目的分析鱼腥草种质资源在分子水平上的遗传多样性。方法应用RAPD技术对92份鱼腥草种质资源进行检测。结果34个随机引物中有32个引物(94.1%)扩增产物具多态性。34个引物共得到200条扩增DNA片段,其中93.5%的片段具多态性。每个多态性引物平均可扩增出5.8个多态性片段。峨眉蕺菜和蕺菜种内平均遗传相似系数(genetic similarity,GS)分别为0.521和0.572,二者种间GS值为0.517。峨眉蕺菜与蕺菜中染色体数目为36的细胞型间相似程度最高,其平均GS值达0.530。栽培蕺菜类群比其野生类群遗传多样性相对较高。聚类分析表明,利用RAPD技术可将全部供试材料区分开,所有材料共划分为14类。其中,绝大多数(62个)聚为一类,且根据RAPD遗传相似系数划分的类群同地理分布有一定关系。结论(1) 鱼腥草种质资源在分子水平上确实存在较大遗传差异。(2) RAPD标记可作为构建鱼腥草DNA指纹图谱的有效工具。(3) 鱼腥草药材道地性与环境因素有关,但更大程度上由其遗传因素所决定。     

硫磺熏蒸时间对白芍中芍药苷含量影响

目的考察硫磺熏蒸白芍的时间对其芍药苷含量的影响。方法采用高效液相色谱法分析白芍经硫磺熏蒸后芍药苷的含量随时间的变化规律。Hypersil ODS色谱柱（4.6 mm×250 mm,2.5μm）;流动相为乙腈-0.1%磷酸溶液（14∶86）;检测波长为230 nm;柱温25℃。结果不同硫熏时间对白芍中芍药苷的影响很大。结论以未经硫熏的白芍的芍药苷含量最高,且随硫磺熏蒸时间的增加芍药苷含量逐渐下降。     

鸡骨草胶囊质量标准研究

目的建立鸡骨草胶囊的质量（白芍、人工牛黄、猪胆汁、栀子、三七等）标准。方法采用薄层色谱法鉴别本制剂中的白芍、人工牛黄、猪胆汁、栀子、三七,采用HPLC法测定本制剂中的栀子苷、芍药苷含量。结果薄层色谱可鉴别出白芍、人工牛黄、猪胆汁、栀子、三七的特征斑点。HPLC法测定的栀子苷、芍药苷分别在0.154-1.54 g（r=0.9982）和0.154-1.54 g（r=0.9999）范围内线性关系良好,平均回收率分别为98.5%、100.1%,RSD分别为0.78%（n=5）、1.5%（n=5）。结论本文方法简便可靠,专属性强,重现性好,可用于该制剂的质量控制。     

环境因子对药用植物木榄遗传多态性及其分布的影响

目的探讨环境因子变化对木揽的DNA遗传多态性的影响。方法利用RP—PCR技术分析中国南部3省6个红树群落中未榄的DNA遗传多态性,并与6个群落环境因子数据对比。结果在扩增到的1277个DNA片段中,25．5％的DNA片断呈现多态性变化：聚类分析显示,来自于山口、高桥、东寨港以及三亚红树群落的木榄可襞为一支（群落问遗传相似系数大于0．90）。文昌和特呈岛木揽分别属于另外两支（与上述4个红树群落的遗传相似系数小于0．90,二者间的遗传相似系数小于0．85）;环境因子数据的聚类分析显示,这6个红树群落可分为4支,其中东寨港和文昌为一支,高桥和特呈岛为一支,山口以及三亚分别属于另外2支。结论地理位置的差异是导致木揽遗传水平出现多态性变化的重要原因之一。     

Choice of Methodology for Assessing Genetic Impacts of Environmental Stressors: Polymorphism and Reproducibility of RAPD and AFLP Fingerprints

PCR-based multi-locus DNA fingerprints represent one of the most informative and cost-effective measures of genetic diversity and are useful population-level biomarkers of toxicologic and other anthropogenic impacts. However, concerns about reproducibility of DNA fingerprints have limited their wider use in environmental biology. We assessed polymorphism and reproducibility of two common fingerprinting techniques, RAPD (randomly amplified polymorphic DNA) and AFLP (amplified fragment length polymorphism), in pedigreed populations of rainbow trout (Oncorhynchus mykiss) to derive general rules for selective removal of problematic fingerprint bands. We found that by excluding bands that comprised less than 1% of total intensity, and by excluding the largest and smallest 10% of the bands, we could achieve nearly 100% reproducibility of AFLP fingerprints. Similar application of band exclusion criteria to RAPD fingerprints did not significantly enhance their reproducibility, and at least 15% of RAPD bands were not fully repeatable, heritable, or transmittable. The RAPD technique produced more polymorphic fingerprints than AFLP; however, considering that a substantial proportion of RAPD markers did not demonstrate Mendelian inheritance patterns, the AFLP methodology is to be preferred for future research.