Genetic variations among mulberry genotypes (Morus alba) as revealed by Random Amplified Polymorphic DNA (RAPD) markers

Turkey is one of the most important mulberry fruit producers in the world. In particular, in Eastern and Inner part of Turkey, mulberry fruits are processed into several products such as `Mulberry Pekmez', `Mulberry Pestil', `Mulberry Kome' etc. Therefore, mulberry fruits give higher economic returns thus have greater employment potentials in Turkey. In order to improve the yield and fruit quality through breeding, the genetic variations and relationships among 15 selected white mulberry genotypes (Morus alba L.) were evaluated using Random Amplified Polymorphic DNA (RAPD). A total of 101 DNA markers were generated by 16 random primers, with an average of 6.3 easily detectable markers per primer. Several RAPD markers showed unique patterns of mean frequency that differed among the white mulberry genotypes. The distance matrix showed that the highest genetic distance (0.902) was between EMS13 and EMS8 and the least (0.413) was between EMS12 and EMS15. According to the results, RAPD analysis can be used for the characterization and grouping of mulberry genotypes. The genetically divergent genotypes identified from this study would be of much use in the future breeding program.     

Random Amplified Polymorphic DNA Diversity among Surface and Subterranean Populations of Asellus aquaticus (Crustacea: Isopoda)

The ecological and evolutionary processes leading to isolation and adaptation of cave animals compared to their surface ancestors are not yet unequivocally understood. In this study the genetic relations of four cave and three surface population of the freshwater crustacean Asellus aquaticus in the Karst region of SW Slovenia and NE Italy were assessed using RAPDs as genetic markers. The results suggest that specialized populations from two caves invaded their subterranean habitat independently, and that their morphological similarity is a result of convergent evolution. Another, less specialized cave population seems to originate from a later colonization of a cave system already inhabited by a more specialized population, but the two populations do not interbreed. This series of temporally and spatially independent invasions has generated a diversity hotspot of non-interbreeding populations of a ubiquitous freshwatercrustacean, which is uniform over most of its range. Genetic variability estimated by the percentage of polymorphic RAPD fragments was similar (86–91%) in most cave and surface populations. Substantially lower values (as low as 49%) were found in two cave populations affected by heavy pollution. Two a priori groupings of populations, traditional subspecies and hydrologically connected groups, were rejected as not significant by means of nested analysis of molecular variance (AMOVA). On the other hand, groupings revealed by UPGMA clustering displayed a significant component of among-group variance. An analysis of gene flow between populations using estimated migration rates confirmed these findings.     

Use of Random Amplified Polymorphic DNA Markers for Mapping the Chickpea Genome

Three interspecific crosses were developed using Cicer arietinum (ICC 4918) as the female parent and wild Cicer species [C. reticulatum - JM 2100, JM 2106 and C. echinospermum - ICCW 44] as the male parent. Cicer arietinum (ICC 4918) × C. reticulatum (JM 2100) cross produced the largest number of F₂ plants and was chosen for linkage mapping using Random Amplified Polymorphic DNA (RAPD) primers. A partial linkage map was constructed based upon the segregation of 36 RAPD markers obtained by amplification using 35 primers. The linkage map consists of two linkage groups with 17 linked markers covering a total of 464.9 cM. Analyses also revealed association of three morphological traits with linked RAPD markers. Out of seven morphological traits tested for association with linked markers in the segregating plants, four Quantitative trait loci (QTL) were detected for the trait leaf length and three QTLs each for the traits leaf width and erect plant habit.     

利用RAPD技术分析实验用比格犬的遗传背景

目的　运用RAPD技术对实验用比格犬 (Beagle)进行遗传背景分析。方法　筛选的 16个RAPD引物对 4 0个样品的分析 ,共得到 93个扩增条带。结果　所有样品的相似系数 80 4 %到 10 0 %间变动 ,平均相似系数为93 2 8% ,聚类分析得到了这些个体的同源树资料。结论　本遗传背景资料可以为以后的比格犬繁育生产和生物医药学的研究应用提供技术指导     

中国紫花苜蓿地方品种随机扩增多态DNA的研究

运用ＲＡＰＤ技术,对１１个样本紫花苜蓿（Ｍｅｄｉｃａｇｏ ｓａｔｉｖａ）的遗传多样性进行了研究。计算了１１个样本之间的遗传相似度和遗传距离,并进行了聚类分析,探讨了它们之间的亲缘关系。结果如下：１．用４３个随机引物对紫花苜蓿的１１个样本的核ＤＮＡ进行了ＲＡＰＤ扩增,共检出４４０条扩增片段,多态性片段３３１条,占７０．６８％。反映了１１个样本之间的遗传变异;２．４３个随机引物对１１个样本扩增出２～１     

波尔山羊杂交后代及其亲本随机扩增多态DNA研究

利用随机扩增多态DNA技术研究了波尔山羊、唐山奶山羊、青龙本地山羊以及波尔山羊与这两个山羊群体杂交后代共计128个山羊个体的随机扩增多态DNA。结果表明：（1）总群体平均遗传多样性指数（Hsp）为0.6974,群体遗传分化指数为0.9706,山羊群体间平均遗传距离指数（0.1314～0.2052）明显大于群体内的相应值（0.0582～0.1440）,上述结果说明,所研究山羊群体不仅具有较为丰富的遗传多样性,而且其核基因组遗传变异主要存在于群体间。（2）山羊群体间的分子聚类关系与各群体间的亲缘关系基本一致。 Abstract:The random amplified polymorphic DNA（RAPD）of 128 individuals was studied,which were from Boer goat, Tangshan dairy goat,Qinglong native goat and their hybrids crossbred with Boer goat.The average index of genetic polymorphism for whole population（Hsp）and the index of genetic differentiation were 0.6974 and 0.9706,respectively.The average index of genetic distance between populations（0.1314～0.2052）was significantly higher than that within populations（0.0582～0.1440）.All of these indicated that the genetic polymorphism was not only abundant,but also the genetic variation was mainly existed between goat populations.The molecular dendrogram among goat populations was in accord with their genetic relationship.     

Intraspecific Variability within Globodera tabacum solanacearum Using Random Amplified Polymorphic DNA

Random amplified polymorphic DNA (RAPDs) were used to investigate the intraspecific variability among 19 geographic isolates of Globodera tabacum solanacearum from eight counties in Virginia and one county in North Carolina. Globodera tabacum tabacum, G. t. virginiae, and the Mexican cyst nematode (MCN) were included as outgroups. Six primers were used and 119 amplification products were observed. Each primer yielded reproducible differences in fragment patterns that differentiated the isolates and species. Hierarchical cluster analysis was performed to illustrate the relatedness among isolates and species. The average Jaccard''s similarity index among isolates of G. t. solanacearum was 74%, possibly representing greater variation than that reported in the literature across different pathotypes of the potato cyst nematode, Globodera pallida, in studies where RAPD were also employed. The RAPD markers described here may be useful for the development of specific primers or probes that could improve the identification of TCN populations. Such improvements in the characterization of TCN genotypes would facilitate the effective deployment of existing and future resistant cultivars to control these economically important pests.     

中国柱花草炭疽病原菌遗传多态性的RAPD分析

在对中国柱花草炭疽病进行广泛调查和病原采样收集的基础上,利用RAPD分子标记技术对43个代表性菌株进行了基因组DNA分析,并与276份国外菌株进行了综合聚类分析。 结果表明所用8个引物的扩增片段位于0.3～2.8kb之间, 菌株间呈现显著的DNA多态性。以柱花草起源中心——南美的柱花草炭疽菌分类为基础,中国柱花草炭疽菌可划分成3大类型即Ⅱ、Ⅲ、Ⅵ类。中国菌株与来自柱花草起源中心——南美的菌株相比之下,其生物多样性和遗传变异性则相对简单。就中国菌株而言海南菌株与广西、广东菌株相比多样性较丰富, 中国柱花草胶孢炭疽菌正在出现种内遗传分化。 从聚类结果看,通常来自于同一个地理区域或同一个寄主基因型的菌株聚成一类, 即同一RAPD聚类组内的菌株通常来自于同一寄主基因型或同一地理区域。说明来自不同寄主基因型或物种的炭疽菌在遗传基因上具有专化性,而地理上隔离的国家或地区的柱花草炭疽病原菌各自具有相对独立的进化途径。     

Genetic diversity and phylogenetic relationships in alder,Alnus firma, revealed by AFLP

Molecular markers for alder,Alnus firma Sieb. et Zucc, have not been studied extensively. Here, we used amplified fragment length polymorphism (AFLP) to investigate genetic relationships among 15 natural populations. EcoRI-ACG + Msel-CTG combinations revealed the highest polymorphism (62.2%). A total of 171 DNA fragments were identified. On average, 58.1% of the AFLP markers that were generated using four primer pairs were polymorphic. Diversity was insignificant among the populations. The combination of a wind-pollinated, outcrossing breeding system along with large population sizes, and the ability to regenerate by stump sprouting may explain the high level of genetic diversity within this species. The majority (98%) of the genetic variance resided within populations. The average number of individuals that were exchanged between populations per generation was very high (N _em = 12.3). Gene dispersal in alder is apparently by seed dispersalvia water and human activity as well as through pollen. Five individuals per population were claded in the same cluster.     

冬虫夏草(Cordyceps sinensis)的随机扩增多态DNA及其遗传分化

本文对来自青藏高原3个区域5个具有代表性地方的13个冬虫夏草(Cordyceps sinensis[Berk.] Sacc.)样本进行随机扩增多态DNA(RAPD)分析。19个随机引物获得的RAPD谱带清晰并呈现多态,单个引物获得的RAPD片段数在3～10个之间。该19个引物在每个样本中扩增的RAPD片段总数平均约为65个。基于遗传距离分析,受试的13个冬虫夏草样本中,来自同一地方的样本间遗传差异甚微,同一区域不同地方的样本间遗传差异较大,不同区域的样本间遗传差异最大。这说明冬虫夏草地理群体间存在着遗传分化。应用UPGMA和NJ方法构建的分子系统树显示,来自5个地方的冬虫夏草实际上可以归并为显著不同的3个组,对应于样本来源的3个区域,提示RAPD标记在冬虫夏草群体中有显著的地区特异性。我们的结果还表明,冬虫夏草地理群体间的遗传差异度与地理距离呈正相关。因此,RAPD作为有效的遗传标记,可用于研究冬虫夏草的遗传多样性、起源以及系统演化等。     

用RAPD技术对罗非鱼遗传变异的研究及应用(英文)

应用随机扩增多态性ＤＮＡ（ＲＡＰＤ）技术检测了一个奥利亚罗非鱼（ａｕ）和湘湖（ｎｘ）、美国（ｎｍ）、沙市（ｎｐ）三个尼罗罗非鱼养殖群体（Ｔａｂｌｅ１）。在２０个引物（Ｔａｂｌｅ ２）中筛选到１２个引物,它们的扩增产物显示了罗非鱼和尼罗罗非鱼二者在群体内或群体间存在遗传差异。其中（Ｆｉｇ．１）, ＯＰＺ０６、 ＯＰＺ１６、 ＯＰＺ１２和 ＯＰＺ１９四个引物分别有一个扩增片段具有种的特异性。它们的大小分别是９００、１５００、１７００和７３０ｂｐ。可以作为鉴别罗非鱼和尼罗罗非鱼二者的分子遗传标记。湘湖（ｎｘ）、美国（ｎｍ）和沙市（ｎｐ）三个尼罗罗非鱼群体内遗传变异相似性指数Ｓ分别为 ０．７９８、 ０．７９５和０．８２４（Ｔａｂｌｅ ３）。表明：这三个尼罗罗非鱼群体都保留了较高水平的遗传变异。而奥利亚罗非鱼（ａｕ）的群体内遗传变异最小。奥利亚罗非鱼（ａｕ）与湘湖（ｎｘ）、美国（ｕｍ）、沙市（ｕｐ）三个尼罗罗非鱼群体之间的遗传距离分别是０．２８５、０．２６２和０．３４４（Ｔａｂｌｅ ３）,说明奥利亚罗非鱼（ａｕ）和沙市尼罗罗非鱼（ｎｐ）杂交将可能产生较强的杂种优势。     

中国貉随机扩增多态DNA及其亚种分化关系

对来自陕西、云南、越南、安徽和广西等地的８只中国貉（Ｎｙｃｔｅｒｅｕｔｅｓｐｒｏｃｙｏｎｉｄｅｓ）进行随机扩增多态ＤＮＡ分析。应用２８个１０ｂｐ的随机引物,平均每只貉获得的ＲＡＰＤ标记数约为１３０条。遗传距离计算结果显示,中国貉个体间的平均遗传距离指数值为１１．２０％,最大值为１４．９３％,最小值为２．９４％。以赤狐（Ｖｕｌｐｅｓｖｕｌｐｅｓ）为外群,应用ＰＨＹＬＩＰ３．０计算软件包中的ＵＰＧＭＡ和ＮＪ聚类方法构建分子系统树。结果表明,不同地理群体间的中国貉存在遗传分化;中国貉可分为４组：（１）广西貉,（２）安徽貉,（３）陕西貉,（４）云南貉和越南貉。其中安徽貉和广西貉间的关系稍近,陕西貉则与云南貉－越南貉稍近。对合中国貉的形态分类、地理分布、ｍｔＤＮＡ多态分析以及进化遗传学的观点,认为陕西貉、广西貉和安徽貉可能与云南貉－越南貉具有等同的分类地位。     

Relationships between Bambusa species (Poaceae,Bambusoideae) revealed by random amplified polymorphic DNA

RAPD (random amplified polymorphic DNA) molecular markers were used to investigate relationships between a sample of Bambusa species from South Eastern China that have been placed in Bambusa or in several segregate genera, Dendrocalamopsis, Leleba, Lingnania, Neosinocalamus and Sinocalamus by different authors. On the resultant neighbor-joining tree, a thorny core Bambusa cluster was distinguished, as was a Lingnania group, and a cluster of Dendrocalamus species with more capitate inflorescences. However, Leleba was found to be a polyphyletic group in the present study.     

Genetic Diversity and Molecular Markers of Cupped Oysters (Genera Crassostrea, Saccostrea, and Striostrea) in Thailand Revealed by Randomly Amplified Polymorphic DNA Analysis

Genetic diversity and species-diagnostic markers of 5 oysters in Thailand, Crassostrea belcheri (Sowerby, 1871), Crassostrea iredalei (Faustino, 1932), Saccostrea cucullata (Born, 1778), Saccostrea forskali (Gmelin, 1791), and Striostrea (Parastriostrea) mytiloides (Lamarck, 1819), were investigated by randomly amplified polymorphic DNA (RAPD) analysis. In a total, 135, 127, and 108 genotypes were observed from primers OPA09, OPB01, and OPB08 (Operon Technologies Inc., kits A and B), and 131 and 122 genotypes from primers UBC210 and UBC220 (University of British Columbia), respectively. Two hundred fifty-four reproducible and polymorphic fragments (200–2500 bp in length) were generated across the 5 investigated species. The average number of bands per primer varied between 12.4 and 32.2. The percentage of polymorphic bands within Crassostrea (53.23%–77.67%) was lower than that within Saccostrea and Striostrea oysters (86.21%–99.36%). Nine, species-specific markers were found in C. belcheri, 4 in C. iredalei, and 2 in S. cucullata. The mean of a ratio between the number of genotypes generated by each primer and the number of investigated specimens of C. belcheri (0.58) was lower than that of the remaining species (0.90–1.00). Genetic distances between pairs of oyster samples were between 0.105 and 0.811. A neighbor-joining tree indicated distant relationships between Crassostrea and Saccostrea oysters, but closer relationships were observed between the latter and Striostrea mytiloides. Received June 6, 2000; accepted September 12, 2000     

Molecular evidence of hybridisation between the endemic Phoenix canariensis and the widespread P. dactylifera with Random Amplified Polymorphic DNA (RAPD) markers

We used RAPD markers to test whether morphologically intermediate individuals between the Canarian endemic Phoenix canariensis and the widespread P. dactylifera correspond to hybrids. Consistent with previous allozyme evidence, the scarcity of appropriate RAPD markers to distinguish P. dactylifera and P. canariensis indicated a close genetic relationship among these species. Only two of the 54 ten-mer primers (OPM-8 and OPK-14) tested in 221 individuals from 7 localities in different islands enabled us to unambiguously identify both species. While P. canariensis possesses two exclusive monomorphic bands of 1000 bp and 750 bp (for OPM-8 and OPM-14, respectively), dactylifera is characterised by two bands of 900 bp and 950 bp for the same primers. The additivity of these taxon-specific bands in the individuals that were morphologically intermediate provided, for the first time, firm evidence for their hybrid origin. Because these hybridisation capabilities pose clear threats to the survival of the endemic P. canariensis and some individuals that had been morphologically characterised as pure P. canariensis revealed later a hybrid nature in the RAPD analysis, we suggest that RAPD markers be used to estimate the possible incidence of introgression in the scarce extant natural populations of P. canariensis. This procedure will provide a straightforward means to select target populations to implement the in situ conservation strategies suggested previously on the basis of allozyme research.We thank Michel Ferry for providing samples of Phoenix dactylifera from the Hort del Gat Research Station on Date Palm and Arid Land Farming Systems in Elche (Spain). The authors wish to thank Jaime OShanahan for his wise comments about Phoenix canariensis. This research was funded by the Gobierno de Canarias (94-2614). A PhD Research Fellowship (Direccion General de Universidades, Gobierno de Canarias) to Gonzalez-Perez. is also acknowledged.     

Analysis of the genetic diversity and relationships among and within species of Hippophae (Elaeagnaceae) based on RAPD markers

RAPD markers were used to assess the genetic diversity and inter- and intra-specific relationships of the genus Hippophae L. and to study the correlation between genetic distances and geographic distances among populations of H. rhamnoides ssp. sinensis. The results analyzed by the percentage of polymorphic loci and Shannon information index indicated that a high level of genetic diversity existed both among and within species of the genus Hippophae. In the UPGMA dendrogram, the species or subspecies were clustered into two main groups but not strictly grouped according to sect. Hippophae and sect. Gyantsensis Lian. The multiple regression analysis and Mantel test both indicated a significant correlation between genetic distance and altitude distance among populations of H. rhamnoides ssp. sinensis, and the cluster analysis suggested that the genetic variation among populations of H. rhamnoides ssp. sinensis was linked to their monophyletic origin. Moreover, some degree of genetic differentiation was found among samples collected at different times.     

赣中南花猪随机扩增多态DNA与群体遗传关系的研究

用RAPD技术检测了泰和冠朝猪、永丰藤田花猪、瑞金三花猪、兴国茶园猪、上犹花猪、万安花猪、乐安花猪等7个赣中南花猪地方类群基因组混合DNA的多态性,经80个随机引物扩增筛选,16个引物产生了共44个多态标记。遗传距离指数计算结果显示：泰和冠朝猪和瑞金三花猪亲缘关系最近,而兴国茶园猪与乐安花猪遗传距离最远。各类群的UPMGA和NJ聚类分析结果与采用生化、免疫和细胞遗传标记分析的结论不尽相同,在此基础上,结合现行分类方法,认为各受试猪群可暂时归并为赣中南花猪类型,但对泰和冠朝猪和万安花猪的品种归属性应作进一步分析。 Abstract: Random amplified polymorphic DNAs (RAPDs) were used to investigate bulked genomic DNA polymorphism of seven pig populations including Taihe Guanchao,Yongfeng Tengtian Spotted,Ruijin Spotted,Xingguo Chayuan,Shangyou Spotted,Wanan Spotted,and Lean Spotted pig.Eighty random primers were screened,of them,16 primers generated polymorphic markers with total number of 44.The genetic distance index matrix indicated that the genetic relationship between Taihe Gaunchao and Ruijin Spotted pig population was the closest,while that between Xingguo Chayuan and Laan Spotted pig population was the farthest.Our molecular phylogenetic trees constructed by UPMGA and NJ methods were in general consistent with that based on biochemical,immunogenetic and cellular genetic markers except for differences in some populations.It was inferred from this study that all pig populations examined could be temporary classified into one breed,however,the population genetic character of Taihe Guanchao and Wanan Spotted pig populations should be further evaluated.     

Restriction Fragment Length Polymorphism and Random Amplified Polymorphic DNA Analysis of Chickpea Accessions

Genetic diversity analysis was carried out in chickpea accessions using restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) techniques. RFLP analysis using 26 Pst I sub-genomic clones on ten chickpea accessions in 130 probe-enzyme combinations detected polymorphism with only two clones. Pst I clones, CG 141 detected polymorphism in ICC 4918 and Pusa 209 while CG 500 detected polymorphism in Pusa 261, ILC 26 and in ILC 13326. These clones detected very few polymorphic markers. Analysis using 10 Eco RI clones on twelve chickpea accessions have shown better hybridisation signal and one clone detected polymorphism in Pusa 256. RFLP analysis of both cultivated and wild Cicer species using heterologous DNA probe Cab3C revealed polymorphism only in wild Cicer species (Cicer reticulatum L., JM 2100). RAPD analysis of 13 chickpea accessions which includes mutants of C 235 and E100Y showed greater degree of polymorphism with 1 - 5 unique DNA bands for all the accessions. Phylogenetic analysis of the RAPD data helped to group the accessions. C 235 and its mutants were found to be closely grouped while E100Y and its mutant E100Ym grouped apart. Desi and kabuli chickpea accessions however, could not be separately grouped. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic diversity and genetic relationships in Hyacinthaceae in India using RAPD and SRAP markers

Genetic diversity and relationship among three genera namely Drimia, Dipcadi and Ledebouria of Hyacinthaceae in India was studied using RAPD and SRAP markers. Twenty one RAPD primers and nine SRAP were used for analyzing 41 accessions. RAPD gave an average 12.6 markers per primer, while SRAP generated 10.1 markers per primer pair. The family emerged very diverged with high polymorphism. The study resolved the three genera into monophyletic groups corresponding to three subfamilies; Urginoideae, Hyacinthoideae and Ornithogaloideae. Drimia wightii emerged a very distinct species and species specific markers were obtained with both marker systems. AMOVA analysis also revealed the genera to be quite well diverged. The two markers showed high correlation (r = 0.932) in Mantel matrix crresspondance test. The combined data also showed a very good correlation with the respective markers individually.     

Taxonomic Study of Dimocarpus longan by Random Amplified Polymorphic DNA Technique

The classification and relationship among 35 samples of longan (Dimocarpus longan Lour. ) have been studied by RAPD (random amplified polymorphic DNA) and statistical cluster analysis. These results showed that it was unlikely possible that "Lizhiben" and "Lizhi kongyan" were hybrid progeny of longan and litchi ( Litchi chinensis Sonn. ) as what some people believe. The "Lizhiben", and "Nanhu Jiaohe" were highly mutant varieties. There were some variance among the "Jiaohe" wilted-nut longan cultivars. Different cuhivars given the same name, such as "Dongbf', "Honghezi" and some other cultivar, were also possible. It was concluded that longan cultivars which have been tested in this research could be classified into 6 groups. The first group, including 12 cultivars such as "Shuizhang", "Youtanben", "Wulongling" etc., which were the well known best cultivars appropriate for producing dry longan, was named as Shuizhang-Wulongling group. The second group, including 11 cultivars like "Chike", "Jiaoyan", "Fuyan", "Chushuben" etc., was named as Chike-Chushuben group. The Dongbi series cuhivars were named after their cultivars as the Dongbi group. The special cuhivars, "Lidongben", "Nanhu Jiaohe" and "Lizhiben" were named respectively as Lidongben group, Nanhu Jiaohe group, and Lizhiben group, because of their more characteristic DNA polymorphism tested in all respective cuhivars. The results indicate that the application of RAPD technique in the taxonomic study has gained some resolution which could not be obtained by morphological analysis and has provided available data for developing longan production and for breeding new longan cultivars.