湖南省饭豆地方种质资源遗传多样性的RAPD和ISSR分析

为揭示湖南省饭豆地方品种的遗传多样性,促进种质资源的有效保护和利用。本研究利用RAPD和ISSR标记分析了源于湖南省各地36份古老饭豆地方品种的遗传多样性,并采用UPGMA方法进行了遗传聚类分析。结果表明：12个RAPD引物和4个ISSR引物在供试材料中共扩增产生81条带型清晰的条带,平均每个引物扩增产生5.06个条带;其中59个为多态性条带,占总扩增条带的72.84%,具有多态性引物的PIC值在0.397~0.968之间;通过UPGMA法进行聚类分析表明供试材料的遗传相似系数在0.605~0.877之间,供试材料被分为4大类,且聚类结果表现明显的地域特征。本研究结果可为本省饭豆种质资源保护和育种研究提供理论依据。     

石斛种质资源遗传多样性的RAPD分析

利用RAPD技术对10种石斛种质资源的遗传多样性及亲缘关系进行了分析。从100条10bp的RAPD引物中筛选获得17条多态性引物,对石斛属的10个种的基因组DNA进行扩增。共获得200条多态性带,平均每个引物产生11.8个多态性条带。材料间遗传相似系数变化范围为0.356~0.676。根据RAPD标记的结果,采用UPGMA法进行聚类分析,将石斛属的10个种区分开来,划分为4类。结果表明：RAPD标记技术较好地从分子水平上揭示石斛种质资源的遗传背景、亲缘关系。     

贵州地方稻种资源的ISSR遗传多样性评价

利用ISSR标记对27份贵州地方稻种资源进行了遗传多样性鉴定与评价。结果表明,用筛选出的12条引物进行PCR扩增,共扩增出清晰、重复性好的谱带155个,其中127个具有多态性,比率为81.9%。ISSR指纹图谱表明,3个引物就能有效地将27份稻种资源完全区别;聚类分析表明,供试品种间的遗传相似性系数为0.555～0.942。在相似性系数为0.66时可将供试种基本按糯性与非糯性分为两大类群。     

RAPD标记对25份柑桔资源及其芽变系的鉴定和多样性分析

利用随机扩增多态性DNA（RAPD）技术,对几份柑橘资源及其芽变系共计25份材料进行了鉴定和遗传多样性分析,从70个引物中筛选出多态性好的10个引物组成单引物和双引物对基因组进行扩增,共扩增出142条谱带,其中多态带72条,共同带70条,平均每个单引物出现多态带6.8条,每对双引物出现多态带1.3条。利用UPGMA法分析发现,这些基因型间的遗传相似系数在0.834～0.966之间,在此基础上,构建了各基因型的遗传关系树状图。结果表明RAPD标记可以准确鉴定出各柑橘资源及其芽变系,这25份柑橘资源代表了25个基因型。较高的遗传相似系数揭示了柑橘资源间狭窄的遗传背景。     

8份剑麻种质亲缘关系的ISSR和RAPD分析

为了揭示剑麻栽培品种的遗传多样性,利用ISSR和RAPD分子标记技术对8份剑麻种质的亲缘关系进行分析。结果表明,筛选后选用的8条ISSR引物和8条RAPD引物,分别产生了53条和66条扩增条带,其中多态性条带分别为44条和61条,多态性条带百分率分别为83.02%和92.42%。根据2种标记的扩增结果,用UPGMA法对8份剑麻种质进行聚类分析,供试材料之间具有较高的遗传多样性,其品种间遗传相似系数分别为0.59~0.80和0.52~0.76。2个标记的聚类结果基本一致,但有点差异,可将供试的8份剑麻种质划分为2类群,而且2个标记聚类结果呈显著相关性,相关系数为0.70。可见,剑麻种质资源的遗传多样性丰富。     

The effect of the number of RAPD markers on the evaluation of genotypic distances in Vitis vinifera

The effect of the number of RAPD fragments on the estimation of genetic distances has been studied on 10 genotypes (Vitis vinifera) using 320 primers. A total number of 1683 scorable bands was obtained but only polymorphic bands (932) were considered in this work. The variation of the genetic distances, in relation to an increasing number of bands, was evaluated through a bootstrap sampling procedure. The mean coefficient of variation (CV) of the genetic distances decreased as the number of bands increased. A CV of 10% was obtained with about 100–150 bands while a CV of 5% with a large number of bands (400–500 and over). To reduce the cost and the time of the analyses, primers with different levels of polymorphism were compared. Genetic distance matrices, based on different numbers of bands and primer polymorphism, were highly correlated with the genetic distance matrix of the whole data set when they were determined with a large number of RAPD fragments, either from high or low polymorphic primers. The effect of the number of bands on the estimation of genetic distances was evaluated also through the variation of the pattern of aggregation among genotypes (cluster analysis). Stable clusterings were detected when a large number of polymorphic bands (400 and over) was used. Some rearrangements of the genotypes in the dendrograms was observed as the number of bands decreased; below 100–150 bands the clusterings were completely rearranged. This revised version was published online in July 2006 with corrections to the Cover Date.     

Random amplified polymorphic DNA analysis of Australian rice (Oryza sativa L.) varieties

Summary The genetic relationships between rice varieties were analysed by using the polymerase chain reaction (PCR), with arbitrary oligonucleotide primers in the random amplified polymorphic DNA (RAPD) method. PCR with 22 arbitrary primers applied to 37 varieties produced 144 useful markers, of which 67% were polymorphic. Thus, with selected primers sufficient polymorphism could be detected to allow identification of individual varieties. Visual examination of electrophoresis gels and analysis of banding patterns confirmed that commercial Australian and USA lines and their relatives were very closely related, with similarity indices of 88–97%. Three varieties originating from more distant geographical centres were easily distinguished, producing variety-specific amplification profiles and expressing a lower similarity index of 80% to all other varieties tested. PCR offers a potentially simple, rapid and reliable method for rice genotype identification and recognition of lines that could contribute genetic diversity to new commercial varieties.Abbreviations PCR Polymerase Chain Reaction - RAPD Random Amplified Polymorphic DNA     

引进国外马铃薯种质资源的遗传多样性分析

应用RAPD标记技术研究国外马铃薯品种的遗传多样性,以期明确这些品种的亲缘关系,为马铃薯优良品种的选育提供理论依据。利用筛选的29条RAPD引物,对24份国外马铃薯品种进行遗传多样性分析。结果显示,29条RAPD引物共扩增出165个条带,其中116条为多态性条带,多态性比例为70.3%;试供的24个国外马铃薯品种间的遗传相似系数在0.514~0.816之间,平均为0.677;相似系数小于0.6的品种对仅为7.25%;24个马铃薯品种可聚为2大类6亚类。结果表明,试供的24个国外马铃薯品种间的遗传相似性高,遗传基础比较狭窄。     

Study of genetic diversity in safflower genotypes using agro-morphological traits and RAPD markers

This research was conducted to study the genetic diversity in safflower (Carthamus tinctorius L.) using agro-morphological traits and RAPD markers. Sixteen selected lines derived from landraces growing in various agro-climatic regions of Iran along with four exotic genotypes were evaluated in a randomized complete block design with three replications under field conditions. Days to emergence, days to initial flowering, days to flowering, days to maturity, plant height, branches per plant, capitula per plant, seeds per capitulum, 1,000-seed weight, seed yield per plant, seed yield, and reaction to powdery mildew (Leveillula taurica Arnaud) were evaluated in this study. Genetic diversity of the genotypes was assessed by RAPD markers. The results indicated significant differences among genotypes for the agro-morphological traits and clustering based on these traits classified the genotypes into five groups. Analysis of the RAPD markers revealed 15 polymorphic primers out of 50 used primers. Based on RAPD data, the highest genetic similarity was observed between the cultivars of “AC Sunset,” “AC Sterling” from Canada and the lowest relatedness observed between a local breeding line “E₂₄₂₈” and genotype “GE₆₂₉₂₃” from Germany. Cluster analysis based on RAPD markers and 54% coefficient of similarity divided the genotypes into five distinct groups. Comparing the clusters based on agro-morphological traits with those from molecular markers showed slight similarities. The finding of high genetic variation for agro-morphological traits and polymorphism at DNA level reveal that agronomic traits can be improved by selection programs.     

Fingerprinting temperate <Emphasis Type="Italic">japonica</Emphasis> and tropical <Emphasis Type="Italic">indica</Emphasis> rice genotypes by comparative analysis of DNA markers

This paper describes the relative efficiency of three marker systems, RAPD, ISSR, and AFLP, in terms of fingerprinting 14 rice genotypes consisting of seven temperatejaponica rice cultivars, three indica near-isogenic lines, three indica introgression lines, and one breeding line of japonica type adapted to high-altitude areas of the tropics with cold tolerance genes. Fourteen RAPD, 21 ISSR, and 8 AFLP primers could produce 970 loci, with the highest average number of loci (92.5) generated by AFLP. Although polymorphic bands in the genotypes were detected by all marker assays, the AFLP assay discriminated the genotypes effectively with a robust discriminating power (0.99), followed by ISSR (0.76) and RAPD (0.61). While significant polymorphism was detected among the genotypes of japonica and indica through analysis of molecular variance (AMOVA), relatively low polymorphism was detected within the genotypes of japonica rice cultivars. The correlation coefficients of similarity were significant for the three marker systems used, but only the AFLP assay effectively differentiated all tested rice lines. Fingerprinting of backcross-derived resistant progenies using ISSR and AFLP markers easily detected progenies having a maximum rate of recovery for the recurrent parent genome and suggested that our fingerprinting approach adopting the ‘undefined-element-amplifying’ DNA marker system is suitable for incorporating useful alleles from the indica donor genome into the genome of temperate japonica rice cultivars with the least impact of deleterious linkage drag.     

Analysis of molecular variance and population structure in southern Indian finger millet genotypes using three different molecular markers

The genetic relationship among 42 genotypes of finger millet collected from different geographical regions of southern India was investigated using random amplified polymorphic DNA (RAPD), inter simple sequence repeats (ISSR), and simple sequence repeats (SSR) markers. Ten RAPD primers produced 111 polymorphic bands. Five ISSR primers produced a total of 61 bands. Of these, 23 bands were polymorphic. The RAPD and ISSR fingerprints revealed 71.3 and 37.4% polymorphic banding patterns, respectively. Thirty-six SSR primers yielded 83 scorable alleles in which 62 were found to be polymorphic. Out of 36 SSR primers used, 14 primers (46.6%) produced polymorphic bands. The SSR primer UGEP7 produced a maximum number of six alleles. Mean polymorphic information content (PIC) of RAPD, ISSR and SSR were 0.44, 0.28, and 0.14, respectively. Molecular variances among the population were 2, 11, and 1% for RAPD, ISSR, and SSR markers, respectively. SSR produced 99% molecular variance within individuals. RAPD and ISSR markers produced a low level of molecular variance within individuals. The STRUCTURE (model-based program) analysis revealed that the 42 finger millet genotypes could be divided into a maximum of four subpopulations. Based on the Bayesian statistics, each RAPD and SSR marker produced three subpopulations (K=3), while ISSR marker showed four subpopulations (K=4). This study revealed that RAPD and SSR markers could narrow down the analysis of population structure and it may form the basis for finger millet breeding and improvement programs in the future.     

49份旱稻种质RAPD标记遗传多样性分析

旱稻具有耐旱、耐贫瘠、适应性广等特点,是稻作育种中的一种特殊的遗传资源。为了更好地利用旱稻种质资源指导稻作育种,采用RAPD分子标记技术对49份旱稻种质（24份海南山栏稻,4份热大系列,21份其他地区旱稻）的遗传基础进行分析。结果表明,利用筛选出的22条多态性较强的引物,构建了49份旱稻种质的RAPD指纹图谱,这22条引物共扩增出375个条带,多态性条带为306条,其多态性为81.60%。参试的49份旱稻种质的遗传相似系数变化范围在0.546~0.952,平均为0.828。聚类分析表明,随着相似系数品种结合线的不同,可将参试的旱稻种质中海南山栏稻与其他地区的种质、大部分旱稻与水旱杂交稻、中国旱稻种质与国外旱稻种质区分开来,第C类海南山栏稻与第D类其他地区种亲缘关系最近,而与第Ⅰ大类亲缘关系较远。多数的品种的RAPD聚类结果与系谱基本吻合。该研究可为稻作种质鉴定及新品种选育等方面提供一定的分子生物学依据。     

温州盘菜地方品种鉴定与指纹图谱的分子标记分析

利用基因组DNA的RAPD、ISSR与SRAP等3种分子标记技术,以日本芜菁品种作为外类群,对来自于温州不同地区具有代表性的10个盘菜品种进行品种鉴定与遗传多样性分析。10个RAPD引物共产生多态性条带70条,多态率为71．7％;12个ISSR引物共产生142条清晰带,其中多态性条带70条,多态率为49．3％;8个SRAP引物组合共产生105条谱带,其中多态性谱带78条,多态性比率为74．3％,表明品种间存在较高的多态性。用单个引物NAURP299、NAUISR43以及SRAP引物组合mel／em2,都可以将11个品种完全区分开来。基于3种标记的聚类分析结果表明,11个材料可以分为3大类,一定程度上能够揭示品种之间园艺学性状的相似性及亲缘关系远近。     

利用RAPD分子标记分析玉米种质遗传多样性

以RAPD分子标记技术对玉米的10个品种的全基因组DNA进行PCR扩增,再用Popgene32软件和SPSS 13.0软件分析扩增结果,研究10个玉米的种质资源遗传关系。结果表明：（1）所选20个引物可扩增出214条RAPD条带;（2）所选引物扩增条带的多态性比率为86.4%,RAPD分子标记扩增10个玉米品种间的相似性系数分别在0.316-0.654之间;（3）用RAPD-PCR扩增条带的分析结果建立了遗传相关系数矩阵、构建了分子树状图、可将10个玉米品种分为3个类群;（4）RAPD分子标记适合于构建玉米的DNA指纹图谱,进行品种鉴定和遗传分析。     

RAPD标记对喀斯特高海拔山区地方玉米自交系遗传多样性研究

本文利用RAPD分子标记技术研究了适应我国喀斯特高海拔山区地方玉米种质的自交系的遗传多样性,从23条引物对我国喀斯特高海拔山区玉米种质的地方自交系对其进行杂种优势群划分.从90个随机引物中筛选出23个多态性好的引物扩增材料,产生出152条多态带,多态性位点比率为86.1%,通过UPGMA聚类分析,将我国喀斯特高海拔山区玉米地方自交系划分为 6个类群.     

RAPD variability in rice (Oryza sativa L.) plants derived from desiccation-tolerant calli

Genetic modification from selfed progenies of 18 rice (Oryza sativa L.) plants regenerated from callus tissues which survived desiccation, were investigated at the DNA level using the random amplified polymorphic DNA (RAPD) method. Twelve 10-mer random primers were used to amplify DNA of progenies from the regenerated plants, and a total of 228 PCR products and 1780 DNA fragments were obtained by primers, generating between four to thirteen major bands. The size of the amplified fragments ranged from 0.2 to 2.55 kb. The results showed that 10 out of 12 primers produced polymorphic bands, two primers (RA31 and RA185) showed no polymorphism among plants tested. A dendrogram of the genetic distance was constructed based on their polymorphism, demonstrating that somaclonal variation exists in rice plants regenerated from callus which survived the desiccation treatment. Part of this variation can be useful in rice breeding. This revised version was published online in August 2006 with corrections to the Cover Date.     

Assessment of genetic diversity in cabbage cultivars using RAPD and SSR markers

Genetic relationship and diversity among seven cabbage cultivars were analyzed using RAPD and SSR markers. These cultivars are of great commercial value in India and are confirmed for their reaction to black rot caused by Xanthomonas campestris pv. campestris. However, so far the extent of genetic diversity and relatedness has not been studied in these cultivars. A total of 17 selected RAPD primers generated 90 bands, 76 of which were polymorphic (84.44%). In addition, 27 selected SSR primers generated 67 amplified bands with 59 of which were polymorphic (87.6%). Though both the marker techniques were able to discriminate the cultivars effectively, analysis of combined data of markers (RAPD and SSR) resulted in better distinction of cultivars. By combining both the markers, a total of 157 bands were detected of which 135 bands (85.98%) were polymorphic, i.e. an average of 5.95 bands per primer. High level of polymorphism (> 85%) recorded with two different marker systems indicated a high level of genetic variation existing among the cultivars. Genetic relationship estimated using similarity co-efficient (Jaccard’s) values between different pairs of cultivars varied from 0.21 to 0.77 in RAPD, 0.42 to 0.82 in SSR, and 0.43 to 0.89 with combined markers. A high correspondence had been recorded between the values of genetic variations generated by UPGMA, clustering, and scatter plot diagrams. The cultivars ‘January King Sel. Improved’ and ‘Golden Acre’ are highly divergent cultivars as demonstrated by both the marker systems.     

DNA polymorphisms in Oryza sativa L. and Lactuca sativa L. amplified by arbitrary primed PCR

Summary Thirty-five rice (Oryza sativa L.) varieties, including 18 japonica, 5 javanica and 12 indica subspecies and 12 lettuce (Lactuca sativa L.) varieties were identified taxonomically, using PCR with originally designed 21 RAPD (Random Amplified Polymorphic DNA) primers and 8 sequence-specific primers, used for amplifying four specific DNA fragments. Use of these primers revealed polymorphisms among varieties in rice and lettuce and facilitates DNA fingerprinting. Dendrograms of both species based on polymorphisms were constructed and genetical relationships were established. In rice, half the number of amplified bands were polymorphic and almost all varieties differentiated. However, differentiation of minor genetic alterations among somaclonal variants or mutants and their mother varieties was not feasible. In L. sativa, 47% of the amplified fragments were polymorphic and all 12 varieties were differentiated. Some of the PCR fragments were variety or type specific, which could be used for indicators for type-selection. The dendrogram obtained showed differentiated clusters of crisphead, leaf and butterhead type, findings in good accord with the classification based on the genetic background.     

30个中国甘薯主栽品种的RAPD指纹图谱构建及遗传变异分析

研究以中国30个甘薯主栽品种为材料,对甘薯RAPD指纹图谱的构建进行了探讨。从102个RAPD引物中筛选出26个多态性丰富的引物用于PCR扩增,共扩增出255条多态性条带,平均每个引物扩增的多态性带数为9.8条。其中S32和S39多态性最高,仅用其中1个引物即能将这30个甘薯品种完全区分开,且由此构建的指纹图谱出现的概率很小,分别为4.77&#215;10^-7（1/221）和1.91&#215;10^-6（1/219）。结果进一步表明,这30个甘薯品种的遗传距离变异幅度较大,为0.0390～0.4306,平均遗传距离为0.3086。RAPD聚类分析表明,地域分布相近的甘薯品种和具有同一亲本的甘薯品种聚在一起,与这些甘薯品种的系谱图一致。     

Genetic Diversity Analysis of Indonesian Aromatic Rice Varieties (Oryza sativa L.) Using RAPD

Journal of Crop Science and Biotechnology - In this research, we investigated genetic diversity of 21 rice genotypes (Oryza sativa L.) assessed using 38 decamer RAPD primers. A total of 405 bands...