DNA Extraction from Eriocaulon Plants and Construction of RAPD System

There have been many arguments on the classification of Eriocaulon Linn. by morphology so far, and little is known about the use of molecular marker for genetic for genetic diversity of Eriocaulon plants. To apply the technique of molecular marker to the research of genetic diversity of Eriocaulon plants, the study of the extraction method of DNA from the Eriocaulon plants and the RAPD system are essential for researchers. In this paper, the extraction of genome DNA from the silica-gel-dried leaves of several species of Eriocaulon distributed in China was studied, and the best RAPD analysis technique condition of Eriocaulon plants was analyzed.

江西省烟草赤星病菌RAPD遗传多样性分析

【目的】为明确江西省烟草赤星病菌Alternaria alternat的遗传多样性,开展了相关研究。【方法】运用RAPD分子标记法对江西省6个主要烟区赤星病分离出的37个菌株进行了遗传多样性、遗传结构与分化及种间遗传变异分析。【结果】其中6个不同地理种群的Nei’s基因多样性指数(He)为0.1618~0.3209,Shannon信息指数(I)为0.2419~0.4785,表明不同地理来源种群之间的链格孢菌具有丰富的遗传多样性;遗传距离分析表明,兴国和信丰的菌株遗传距离最小,亲缘关系较近,石城和黎川的菌株遗传距离最大,亲缘关系较远;居群每代迁移数(Nm)为1.8171,表明不同地理来源的种群间存在一定程度的基因流动;利用NTSYS软件按UPGMA法聚类分析,当相似系数划定为0.56时,可将37个菌株划分为10个类群,37个菌株基本上以地理来源聚类到不同分支,表明菌株间的遗传多样性受其地理来源影响明显。【结论】利用RAPD分子标记法明确了江西省主要烟区烟草赤星病菌Alternaria alternat的遗传多样性,为江西省烟区选育抗赤星病品种及其综合防治提供措施奠定理论基础。

基于RAPD技术对大曲芽孢杆菌近似菌株的分类研究

大曲是浓香型白酒酿制过程中最主要的微生物源,大曲中微生物的类型、丰度、新陈代谢活动等均会对浓香型白酒品质产生很大的影响。为了探究大曲中芽孢杆菌的分类及多样性,利用菌株随机扩增多态性DNA标记(random amplified polymorphic DNA,RAPD)分型和16S rDNA序列分析技术,对已分离出的芽孢杆菌进行RAPD分型和16S r DNA序列分析。结果表明,37株芽孢杆菌中有22株枯草芽孢杆菌、9株蜡样芽孢杆菌、3株地衣芽孢杆菌,其中枯草芽孢杆菌又可细分为8类,蜡样芽孢杆菌可细分为3类,地衣芽孢杆菌可分为3类;RAPD分型可以为大曲中芽孢杆菌的快速分型及后续的分类学研究提供理论依据,为研究芽孢杆菌在白酒酿造中的作用奠定基础。

Investigating the genetic diversity of several varieties of Iranian tomato (Solanum lycopersicum) using RAPD and ISSR molecular markers

Background:Numerous studies have demonstrated the existence of approximately 7,500 genetic tomato varieties worldwide.Hence,it is crucial to assess the genetic diversity among tomato cultivars.This study aimed to investigate the genetic diversity of selected Iranian tomato cultivars(Solanum lycopersicum)using RAPD and ISSR molecular markers.Method:Ten RAPD primers and ten ISSR primers were employed to assess the genetic diversity among 10 tomato cultivars:Matin,RFT 112,Hirad,Golsar,Raha,Hengam,Hedah,Fasa,JS12,and Emerald.Data analysis involved the UPGMA algorithm and NTYSYSpc software.Results:RAPD analysis revealed close genetic proximity between Fasa and JS12,as well as between Raha and Hadieh.Conversely,the RFT 112,Hengam,Hirad,and Emerald cultivars exhibited significant genetic diversity within this group.ISSR primer analysis identified Hengam as the most diverse variety,while Matin,Emerald,and Vibrid,as well as Raha and JS12,displayed genetic similarities with minimal observed diversity.Furthermore,the overall analysis of the cultivars using RAPD and ISSR markers indicated that Hengam exhibited the highest diversity among all the varieties.Notably,Raha and JS12 demonstrated limited diversity in this analysis.Conclusion:This research demonstrates substantial genetic diversity among the investigated tomato varieties,with Hengam displaying the highest diversity within this group.Furthermore,ISSR markers proved more effective in determining genetic diversity in tomato plants.

Optimization of RAPD Reaction System on Genomic DNA of Agrocybe chaxingu Huang

[Objective] The research aimed to screen out optimum RAPD reaction system on genomic DNA of Agrocybe chaxingu Huang.[Method] The single factor experiment was adopted to select the required Mg2+ concentration, template DNA concentration,primer concentration,dNTPs concentration,Taq enzyme concentration and anneal temperature initially.[Result] The optimum reaction system for RAPD amplification of Agrocybe chaxingu Huang was listed as follows:2.5 μl Buffer, 2 mmol/L Mg2+, 75 ng DNA, 0.5 μmol/L primer, 150 μmol/L dNTPs and 2.0 Taq enzyme.The reaction process was also listed as follows: denaturation for 5 min at 92 ℃,35 cycles(1 min at 92 ℃, 1 min for 35.5 ℃ and elongation for 2 min at 72 ℃),10 min at 72 ℃.[Conclusion] The research provided reference for conducting RAPD analysis of and studying genetic relationship and genetic diversity of Agrocybe chaxingu Huang.

Optimization of RAPD Reaction System of Taxus cuspidate Sieb. et Zucc

[Objective] The research aimed to study the optimization conditions of RAPD reaction system of Taxus cuspidata.[Method] Using the leaves of Taxus cuspidata as materials,DNA was extracted by using modified CTAB method.The influences of template DNA concentration,primer concentration,dNTP concentration,Taq DNA polymerase amount and magnesium ion concentration on the amplification effects of RAPD of Taxus cuspidate were analyzed.[Result] Through analysis and comparison of various factors,the optimized reaction system was established as follows：in a total volume of 20 μl,containing 10 ng DNA,2.0 mmol/L Mg2＋,0.2 mmol/L dNTPs,15 pmol/μl random primer,1.0 U Taq polymerase,2 μl 10×buffer,and the supplementary ddH2O.[Conclusion] The research provided technical support for further discussion on the related studies with the sexual identification of Taxus cuspidate.

Using RAPD Method on Systematic Evolution of Four Species in Anura

The phylogenetic relationships of four species,Bufo melanostictus,Hyla chinensis,Rana limnocharis and Rana guentheri,which belong to three differrent families of Anura,were detected with RAPD technique.The genomic DNA of each species was amplified with 19 random primers.16 primers given clear amplified bands were used for analysis and the genetic distances between four species were calculated.The results show that RAPD bands obtained by all 16 primers evinced different degree polymorphisms.The genetic distance between R.limnocharis and R.guentheri is the nearest,that between B.melanostictus and H.chinensis is the second nearest,and that between B.melanostictus and R.guentheri is the furthest.The different distances also indicated that the relationship between Bufoidae and Hylidae is closer than that between Bufonidae and Ranidae at genomic DNA level.In concordant with the conclusion of the morphology,chromosomal and mitochondrial DNA studies,our results provide a new evidence of the systematic evolution of the three families of Anura at DNA molecular level.

重庆养殖中华鳖群体遗传多样性的RAPD分析

[目的]利用RAPD标记技术对重庆4个养殖地区中华鳖的遗传多样性进行分析。[方法]选取(潼南、永川、江津、长寿)4个地区养殖中华鳖样本,运用RAPD方法对4个地区的中华鳖遗传多样性与其遗传距离进行了分析。[结果]用20个随机扩增引物得到142个扩增片段,多态片段数量为95个,4个地区的多态位点比例范围为50.78%~64.54%,总多态位点比例为66.90%,总基因多样性为0.227,种群内基因多样性为0.135,遗传分化系数为0.307,基因流为1.127。4个养殖中华鳖群体的遗传距离为0.049~0.151。[结论]重庆4个地区养殖中华鳖遗传多样性较高,且大部分遗传变异存在于种群内,个体间亲缘关系较近,遗传变异度较小。

Assessment of Genetic Variation in Soybean (<i>Glycine max</i>) Accessions from International Gene Pools Using RAPD Markers: Comparison with the ISSR System

Soybean (Glycine max) is one of the most important crops in the world in terms of total production and usage. It is also among the least diverse species. The main objectives of the present study were to 1) assess the level of genetic variation among soybean (G. max) accessions from different countries using Random Amplified Polymorphic DNA (RAPD) markers and 2) compare Inter Simple Sequence Repeats (ISSR) and RAPD marker systems in detecting polymorphic loci in soybeans (G. max). Genomic DNAs from 108 soybeans (G. max) accessions from 11 different gene pools were analyzed using several ISSR and RAPD primers. The average level of polymorphic loci detected with the RAPD primers was 35%. The soybean accessions from the China, Netherlands, and Canada gene pools were the least genetically variable with 25%, 26%, and 30% of polymorphic loci, respectively. Accessions from Hungary (43%) and France (48%) showed the highest level of polymorphism based on the RAPD analysis. Overall, RAPD data revealed that the accessions from different countries are closely related with 64% genetic distance values below 0.40. The levels of polymorphic loci detected with the RAPD and ISSR marker systems were in general moderate and similar even if they target different regions of the genome. A combination of different marker systems that include RAPD/ISSR, microsatellites (SSR), and SNPs should provide the most accurate information on genetic variation of soybean (G. max) accessions.

基于RAPD分子标记的观赏春石斛遗传多样性分析

【目的】利用RAPD分子标记对观赏春石斛的遗传多样性进行分析。【方法】以10份观赏春石斛种质资源为供试材料,采用100条RAPD随机引物进行筛选。【结果】从筛选出来的15条具有良好多态性的引物中扩增得到125个DNA条带,其中80个条带表现出多态性,多态性位点比率为63.89%。聚类分析结果显示,在相似系数为0.37时10份样本分为两大类,在相似系数0.41时可进一步分为两个亚类群。【结论】利用RAPD分子标记可以鉴别观赏春石斛种质资源。

基于ISSR和RAPD分子标记的15株糙皮侧耳遗传多样性分析

平菇(糙皮侧耳)是我国食用菌栽培主要品种之一,国内各地栽培品种较多,平菇资源的遗传多样性较丰富。为了明确平菇菌株间的亲缘关系及遗传距离,借助ISSR和RAPD两种分子标记技术,对收集到的国内主产区的15个平菇菌株进行DNA遗传多样性分析,为后期平菇适应性试验提供理论依据。通过ISSR和RAPD两种分子标记引物筛选试验,获得9对多肽引物,利用9对引物对15份平菇DNA样品进行多样性PCR扩增,并采用NTSYS-PC分析软件对供试菌株的遗传多样性进行聚类分析。结果显示,参试平菇菌株遗传距离在0.26~0.56;9对引物共扩增产生了102条清晰条带,平均每对引物扩增产生11.3条带;多态性条带90条,多态性条带比例为88%,平均每对引物组合扩增产生10条多态性条带;在遗传距离0.36的水平上,供试的15个平菇品种分为4个类群,P101和P121分别单独分为一个类群,P102、P112、P120、P122、P104、P125、P107、P113、P124归为一个类群,P117、P123、P118、P119归为一个类群。供试材料之间遗传差异大,遗传多样性较丰富,研究结果为本地区平菇产业发展及育种提供新的菌株。

国产姜黄属药用植物RAPD分析与分类鉴定

运用 RAPD技术刻划了国产姜黄属药用植物的遗传背景差异 ,并结合经典分类和异地栽培观察和聚类分析 ,初步澄清了姜黄属植物的一些分类学争议 ,从

当归药材道地性的RAPD分析

目的 从 DNA分子水平上探讨当归药材道地性的生物学基础。方法 对来自不同产地的 18个当归样品 ,提取其基因组 DNA,并进行 RAPD分析。对筛选出的 10个引物的 RAPD结果进行聚类分析 ,得出 18个当归样品的聚合树状图。结果 显示地理分布距离越小 ,当归的遗传差异越小 ;反之 ,越大。但生态环境特别是小生境对当归遗传差异的作用亦不可忽视。

中药白芷种质资源的RAPD分析

目的：对中药白芷的种质资源进行分析。方法：用１２个随机引物对野生白芷、杭白芷的４个居群超过１７个个体的基因组ＤＮＡ进行ＲＡＰＤ分析。结果：共扩增出４０个位点，其中多态位点２６个，占６．５％；祁、杭白芷的多态位点的百分率显示出低水平的遗传变异。根据ＲＡＰＤｉｓｔａｎｃｅ分析，祁、杭间的遗传距离（０．１６０）小于祁白芷居群内单株间和杭白芷不同居群间遗传距离，并且它们都小于野生白芷与祁、杭白芷间的遗传距离。结论：祁、杭白芷应属同一类群，与野生白芷有一定区别。

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.

DNA提纯方法对9种菊属植物RAPD的影响

菊属植物组织中所含的多酚类、单宁、色素及多糖类物质是干扰RAPD扩增反应的主要成份。通过比较试验,使用十二烷基硫酸钠(SDS)法提取,DNA,并补加非水溶性聚乙烯吡咯烷酮(insoluble PVP),有效地去除了上述杂质,经纯化的DNA能很好地用于RAPD分析。试验还发现,4月上旬至5月中旬植物材料处于旺盛的营养生长状态时取材提取DNA较为适宜。

中国对虾黄、渤海沿岸地理群的RAPD分析

采用随机扩增多态 ＤＮＡ（ｒａｎｄｏｍ ａｍｐｌｉｆｉｅｄ ｐｏｌｙｍｏｒｐｈｉｃ ＤＮＡ，简称ＲＡＰＤ）技 术，对中国对虾的黄、渤海地理群的３２个个体遗传多样性进行了检测．使用ＯＰＩ系列 的２０个 １０ ｂｐ的寡核苷酸随机引物，对每个个体的基因组 ＤＮＡ进行扩增，结果为：１７ 个引物获得了谱带清晰且重复性好的产物，扩增片段的分子量在 ２ ０００～２００ ｂｐ之间， 共检测 １０６个位点，其中多态位点数为 ３９个，占 ３６． ８ ％；平均遗传距离为 ０． ０９４１＋ ０． ０２０ ６，有 ６７个标记（占 ６３． ２％）在整个群体的 ３２个个体间表现稳定的一致性。

利用RAPD分析大青杨天然群体的遗传结构

本文利用ＲＡＰＤ分子标记技术从ＤＮＡ分子水平上探测了大青场（ｐｌｐｕｌｕｓｕｓｓｕｒｉｅｎｓｉｓＫｏｍ．）天然群体的遗传结构和分化程度。结果得出：用１４个随机寡核苷酸引物共产生１８０个扩增片段，扩增片断在２１１ｂｐ至１６３６ｂｐ之间。Ｓｈａｎｎｏｎ表型多样度（ＨＯ）估测值在群体间变动范围为０．２７１至０．３９２，平均为０．３１０。对分子水平变异分为群体间和群体内两部分进行分析，群体间分量占总变异的６２．３％，群体内只占３７．７％。不同引物在群体内探测能力也各不相同，ＣＨｌ－ｌ引物探测多样度（ＨＯ）最高（０．５４０），而２１１６引物最低（０．１５１）。

日本对虾野生种群和养殖种群遗传结构的RAPD标记研究

于1997年4月在福建和山东青岛分别采集日本对虾野生种群和养殖群体的样品,采用RAPD标记技术对其遗传结构进行初步研究。用20个随机引物在野生种群和养殖群体中分别扩增出157和153条DNA片段,其中多态性片段数分别为85和58条,多态性片段的比例分别为54.14％和37.91％,平均杂合度分别为0.2517和0.1343。野生种群的多态性位点比例和条合度明显高于养殖群体,说明中国的日本对虾野生种质资源状况较好,应加以合理保护。与野生种群相比,养殖群体的杂合度有所下降,提示在日本对虾的育苗育种过程中应引进标记辅助选择等技术手段。

蜜蜂DNA提取纯化与RAPD反应体系的建立

以意大利蜜蜂为材料,研究了蜜蜂DNA的提取以及对RAPD分析的影响因素,包括模板浓度、Mg2+、dNTP和引物,建立了适于蜜蜂RAPD分析的PCR反应体系,即20μL反应体系中,包括10mmol/LTris-HCl(pH8.3)、50mmol/LKCl、20～60ngDNA、3.0mol/LMgCl2、0.2mmol/LdNTP、0.5μmol/L随机引物和1.5unitTaq聚合酶。扩增程序为:94℃预变性5min,94℃变性1min,36℃退火1min,72℃延伸2min,40个循环;最后在72℃延伸10min。