青蒿素高含量地区黄花蒿种质资源的简单重复序列区间分析

目的:分析广西、贵州和重庆3个青蒿素高含量地区的野生黄花蒿资源的遗传多样性。方法:以40份黄花蒿为试材,通过简单重复序列区间(ISSR)分析,运用POPGENE version1.31软件和TREECONW软件计算相关参数,UPGMA方法聚类,构建亲缘关系树状图。结果:18条ISSR引物扩增出84条带,多态比率为79.76%;物种水平上,ISSR标记揭示的Nei's基因多样性指数为0.2019,Shannon信息指数为0.3244;3个地区之间ISSR标记揭示的Nei's基因多样性指数范围为0.1867～0.1943,Shannon信息指数范围为0.2936～0.3073;ISSR检测不同材料间遗传距离范围为0.1467～0.4493,平均为0.3132。结论:ISSR标记能有效地揭示青蒿素高含量地区野生黄花蒿极为丰富的遗传多样性,可为进一步的选择育种提供更多的种质资源。     

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。     

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。     

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。     

不同居群白芍的DNA随机扩增多态性研究

目的分析6个不同居群白芍的遗传多样性,为白芍的种质鉴定及遗传多样性分析提供依据。方法运用随机扩增多态性DNA(RAPD)技术,对浙江磐安、四川中江、安徽亳州、上海崇明、江苏宿迁和山东荷泽居群白芍的基因组DNA进行随机扩增,利用NTsys2.10e软件计算遗传相似性,运用UPGMA法进行聚类分析并构建树状图。结果共筛选了70个随机引物,从中挑选出8条多态性强、重复性好的引物,共检测出215个位点,多态性位点137个,多态位点比率为63.7%,UPGMA聚类可以将不同来源的白芍很好地区分开。结论不同产地间的白芍存在丰富的遗传多样性,RAPD分子标记方法可以用来鉴定不同产地的白芍。     

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。     

不同地区广金钱草遗传多样性的ISSR分析

摘 要 目的：对广东省、广西壮族自治区共15个产地的广金钱草遗传多样性以及亲缘性关系进行分析。方法: 应用inter simple sequence repeat（ISSR）技术,对广东省、广西壮族自治区的样品进行遗传多样性分析;使用Popgene 1.32软件对扩增结果进行分析,并使用UPGMA软件绘制聚类图。结果: 7条ISSR引物扩增出的产物共有51条清晰的条带,其中多样性条带为41条;Shannon多样性指数(I)为0.3,Nei’s遗传多样性指数(H)为0.246 4,居群间的遗传分化系数(G_ST)为0.123 8,基因流(N_m)为3.539 7。结论:广东、广西两地栽培以及野生广金钱草具有较高的遗传多样性,其遗传距离跟地理距离有一定的关联性。     

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。     

四倍体黄芩及二倍体黄芩遗传差异的ISSR分析

目的 分析研究四倍体黄芩新品系D20和二倍体黄芩之间的遗传差异。方法 采用ISSR分子标记技术对四倍体黄芩及二倍体黄芩之间的遗传差异进行研究。结果 从100条引物中筛选得到9条ISSR引物用于正式扩增,多态性百分率为84.2%;遗传距离的变异范围是0.086 9~0.242 9,聚类分析结果将黄芩样本分为两大类,二倍体聚为一类,四倍体聚为另一类。结论 四倍体黄芩新品系D20和二倍体黄芩存在明显的遗传差异,ISSR分子标记能有效的揭示材料间的多态性。     

22份莲种质资源的ISSR遗传多样性研究

目的：对莲不同品种的遗传多样性和亲缘关系进行分析。方法采用ISSR 标记技术对采自广东、湖南、江西、福建、湖北、浙江、江苏、北京、河北等地的22份不同品种莲的遗传多样性进行了研究。结果从引物库中的100条ISSR引物中筛选出18条多态性稳定、清晰的引物,共扩增出221条带,其中多态性条带数为181条,多态位点百分率为81．9％。 Nei－Li相似系数（ GS）计算方法得到样品间的GS值在0．37～1．00之间。结论22份莲样品间的遗传多样性较高,而地域性差异不明显,此法可用于莲不同品种间的鉴别。     

高效液相色谱法评价贵州金钱草代谢产物槲皮素和山奈素的多样性

目的： 建立金钱草中槲皮素和山奈素的HPLC测定方法,用以测定贵州省不同来源地的21份金钱草中槲皮素和山奈素的含量,并对其多样性进行分析。方法： 采用Alltima C₁₈柱（5 μm,250 mm×4.6 mm）,以甲醇-0.4%磷酸（50∶50,V/V）为流动相,柱温：25℃,流速：1.0 mL·min^-1,检测波长为360 nm;用SPSS 19.0软件对不同来源地金钱草中槲皮素和山奈素进行聚类分析。结果： 槲皮素和山奈素分别在0.028 6~0.196 5 mg （r=0.999 7）和0.047 7~0.381 4 mg （r=0.999 9）的定量范围内,色谱峰面积与进样量呈良好的线性关系,相关系数均大于0.999。槲皮素和山奈素在精密度实验、重复性实验和稳定性试验中的RSD分别为0.24%和0.58%、0.95%和1.32%、1.02%和1.08%,RSD均小于1.40％,加样平均回收率为99.42%和98.36%。不同来源地的21份金钱草中槲皮素和山奈素含量差异较大。结论： 用HPLC法测定金钱草中槲皮素和山奈素含量简单、精确、重复性高,贵州金钱草中槲皮素和山奈素含量差异较大,具有丰富的多样性及一定的地域性。     

Intersimple sequence repeats (ISSR) molecular fingerprinting markers for authenticating populations of Dendrobium officinale Kimura et Migo

Intersimple sequence repeats (ISSR) molecular fingerprinting markers have been employed to authenticate eight populations of Dendrobium officinale using 10 primers selected from 76 ISSR primers. A total of 127 DNA fragments were amplified, of which 115 were polymorphic (90.5% of all bands). Sixteen specific authentication markers have been found. To enhance the efficiency of authentication, ISSR fingerprinting codes have been constructed using six polymorphic bands for authenticating D. officinale populations. Eight wild populations of D. officinale have been authenticated accurately using ISSR.     

西河柳种质遗传多样性的ISSR分析

目的：应用ISSR-PCR标记技术研究中药材西河柳基原植物柽柳及其近缘种的亲缘关系和遗传多样性,为西河柳药材鉴定和育种提供参考。方法：对柽柳7个居群以及甘蒙柽柳、多枝柽柳及购自药材市场西河柳共11个样本进行ISSR-PCR分析, 采用NTSYS-pc软件计算柽柳各居群及近缘种间的Jaccard遗传相似系数, 按非加权配对算术平均法(UPGMA) 建立所研究类群的系统聚类图。结果：14条引物扩增出106个条带,其中多态带为83条,占总扩增条带数的78.3％。从聚类分析图可以看出,柽柳不同居群样本首先聚在一起,表明是一自然物种;甘蒙柽柳与柽柳具有较近的亲缘关系。研究还表明目前商品市场西河柳药材来源较混乱。结论：ISSR标记可以为西河柳原植物及其近缘种的种质资源鉴定、遗传关系分析及栽培育种提供分子生物学依据。     

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.     

Genetic diversity of Paris polyphylla var. yunnanensis, a traditional Chinese medicinal herb, detected by ISSR markers

Paris polyphylla Smith var. yunnanensis (Franch.) Hand.-Mazz. is an important Chinese medicinal herb. Because of overharvesting, the wild populations of this herb have greatly declined and become fragmentized. In this paper, ISSR markers were used to determine the genetic diversity and genetic structure of this variety represented by a total of 153 individuals from three natural populations and three cultivated populations. Fourteen primers produced a total of 251 bands, of which 227 were polymorphic (PPB=90.44%). For the natural populations, the results showed that genetic differentiation was mainly within populations (GST=0.1952), with low genetic diversity at the population level. At the population level, genetic diversity of the cultivated populations was relatively higher than that of the natural populations (PPB=57.24% vs. 53.38%, HE=0.153 vs. 0.151, HO=0.241 vs. 0.235). This pattern can be explained by the recent introduction and artificial selection of cultivars from comparatively wide areas of origin, and subsequent gene flow among populations in cultivation. Although the neighbour-joining cluster analysis seemed to suggest that there was conspicuous genetic differentiation between the natural and cultivated populations, the AMOVA showed that only 4.84% of the total variance existed between groups of natural and cultivated populations, while 67.51% of the variance occurred within populations. In the end, some suggestions for conservation of this important herb are proposed.     

Genetic and metabolic diversity in Stevia rebaudiana using RAPD and HPTLC analysis

Abstract

Context: Stevia rebaudiana Bertoni (Asteraceae) is an important medicinal plant and is much used due to its zero calories sweetening property. Stevia leaves as well as its extracts and pure compounds are currently used in the preparation of several medicines, food products and neutraceuticals.

Objective: To study the genetic and metabolic variability in S. rebaudiana among accessions of different geographical regions of India using random amplified polymorphic DNA (RAPD) markers and high-performance thin layer chromatography (HPTLC) analysis.

Materials and methods: The RAPD analysis of Stevia rebaudiana (11 accessions) was carried out using 20 random operon primers. Dendrogram was constructed for cluster analysis based on the unweighted pair group method with arithmetic means (UPGMA) using Winboot. The HPTLC analysis of all samples was carried out on silica using acetone:ethyl acetate:water (5:4:1, v/v/v) for fingerprinting and quantification of stevioside and rebaudioside A at 360?nm after spraying with anisaldehyde sulphuric acid.

Results: Ten out of 20 primers screened were found most informative; amplification products of the genotypes yielded a total of 87 scorable bands (67 polymorphic), whereas genetic similarity (GS) coefficient (0.01–0.08) and polymorphism (67.24–92.40%) showed huge variability. Similarly, HPTLC analysis showed large variation among different samples with respect to their presence or absence of metabolite and their concentration.

Conclusion: Out of the 11 Stevia accessions, Delhi and Mohali varieties showed much relatedness with each other and were concluded to be the superior genotype in context to RAPD and HPTLC analysis. The information obtained here could be valuable for devising strategies for cultivating this medicinal plant.     

苦荞遗传多样性分析与核心种质筛选--测试文章

利用SSR分子标记技术,对来自西藏、陕西、四川、贵州等4省区的210份苦荞品种进行遗传多样性研究。结果表明,所选用的50对SSR引物中,有16对引物多态性良好,共扩增出178个条带,其中多态性条带达118个,占总数的66.3%;210份苦荞种质的遗传变幅为0.62～0.98,平均值为0.80,在遗传相似系数为0.88处可划分为5大类。聚类结果显示,来自同一地区的苦荞品种显示出聚为一类的趋势,表明苦荞的遗传信息受地理分布的影响较大;第V类所聚的41份分别来自西藏（27份）、陕西（8份）和贵州（6份）的品种表现出较为丰富的遗传多样性,与其他种质相比,这41份材料不仅遗传差异较大,而且遗传来源广泛,可作为苦荞核心种质筛选的基础。