Microsatellite DNA and RAPD fingerprinting,identification and genetic relationships of hybrid poplar (<Emphasis Type="Italic">Populus x canadensis</Emphasis>) cultivars

Microsatellite DNA markers of ten SSR loci and 248 RAPD loci (resolved by 26 RAPD primers) were used for DNA fingerprinting and differentiation of 17 widely grown Populus x canadensis syn. Populus x euramericana (interspecific Populus deltoides x Populus nigra hybrids) cultivars ("Baden 431", "Blanc du Poitou", "Canada Blanc", "Dorskamp 925", "Eugenei", "Gelrica", "Grandis", "Heidemij", "I-55/56", "I-132/56", "I-214", "Jacometti", "Ostia", "Regenerata", "Robusta", "Steckby" and "Zurich 03/3"), and determination of their genetic interrelationships. Informativeness of microsatellite and RAPD markers was also evaluated in comparison with allozyme markers for clone/cultivar identification in P. x canadensis. High microsatellite DNA and RAPD genetic diversity was observed in the sampled cultivars. All of the 17 P. x canadensis cultivars could be differentiated by their multilocus genotypes at four SSR loci, and were heterozygous for their parental species-specific alleles at the PTR6 SSR locus. Except for "Canada Blanc" and "Ostia", which had identical RAPD patterns, all cultivars could also be differentiated by RAPD fingerprints produced by each of the two RAPD primers, OPA07 and OPB15. For microsatellites, the mean number of alleles, polymorphic information content, observed heterozygosity, observed number of genotypes and the number of cultivars with unique genotypes per locus was 5.2, 0.64, 0.67, 5.7 and 2.2, respectively. For RAPD markers, the number of haplotypes per locus, and the number of cultivars with unique RAPD profiles per locus were 1.06 and 0.72, respectively. Overall, microsatellite DNA markers were the most informative for DNA fingerprinting of P. x canadensis cultivars. On the per locus basis, microsatellites were about six-times more informative than RAPD markers and about nine-times more informative than allozyme markers. However, on the per primer basis, RAPD markers were more informative. The UPGMA cluster plots separated the 17 cultivars into two major groups based on their microsatellite genotypic similarities, and into three major groups based on their RAPD fragment similarities. Both the microsatellite and RAPD data suggest that the cultivars "Baden 431", "Heidemij", "Robusta" and "Steckby" are genetically closely related. The inter-cultivar genetic relationships from microsatellite DNA and RAPD markers were consistent with those observed from allozyme markers, and were in general agreement with their speculated origin. Microsatellite DNA and RAPD markers could be used for clone and cultivar identification, varietal control and registration, and stock handling in P. x canadensis.     

Application of RAPD Analysis in the Study on the Origin of Chinese Cultivated Chrysanthemum

The RAPD ( random amplified polymorphic DNA) markers generated by 22 20-ruer primers, discriminated 7 wild Dendraathema species, 14 cultivars and 5 interspecific hybrids by banding profiles. Of 224 bands observed, 34 bands (15%) were polymorphic. Cluster analysis based on UPGMA method indicated that: of 7 wild species, D. indicum, D. vestitum and D. nankingense were closest to the cultivated groups while D. zawadskii was most distinct. Genetic distances between the former three species and cuhivars were less than 0.40 but those between D. zawadskii and cuhivars were more than 0.50. According to the RAPD analysis and previous research, the authors discussed the origin of Chinese cultivated chrysanthemum and proposed that the cuhivars of this genus may derive from D. indicum, D. vestitum and D. nankingense. The results have provided basic research data for study on the origin of Chinese cultivated chrysanthemum.     

低温冷害对龙眼成花的影响及抹除荔枝和芒果冷害花穗促进腋芽再生花穗的效果

为探讨早春极端低温对龙眼成花的影响及芒果和荔枝花穗冷害发生时应对措施的效果,在2008年早春低温冷害时,通过抹除芒果和荔枝的冷害顶生花穗,研究该应对措施对促进腋芽再分化花芽并抽生花序的效果,并在低温冷害后,对不同龙眼品种的成花情况进行调查。结果表明:抹除荔枝冷害顶生花穗后能显著促进黑叶、钦州红荔、糯米糍、立夏红腋芽再生花序,平均单株花穗数分别为139、62.5、28和29穗,分别比对照的高119、22.5、25和26穗;而妃子笑、三月红、桂味和禾荔的处理树和对照树之间差异不明显。抹除芒果冷害顶生花穗后,台农1号、贵妃、桂热82号、红象牙和金穗芒的平均单株成花数分别为92、18、131、20.5和18穗,明显高于对照;而凯特芒、桂热120、吉尔、紫花芒和金穗芒处理树和对照树之间差异不显著。龙眼低温冷害后成花较好的有桂明、储良、石硖、小广眼、大乌圆和大广眼,平均单株花穗数分别为88、67、52.7、52、51和50穗;其次是桂香、乌龙岭、东壁、立冬本和早白露,平均单株花穗数分别为39、26、25、23.5和21.5穗。     

Detection of DNA polymorphism among pea cultivars using RAPD technique

An influence of some Random Amplified Polymorphic DNA (RAPD) reaction factors on resulting banding pattern and the ability of RAPD technique to detect DNA polymorphism among six economically important pea cultivars was tested. Relatively high level of DNA polymorphism among peas was observed, using polyacrylamide/urea gels and silver staining. Altogether 13 arbitrarily designed primers produced 313 amplification products. In addition 59 polymorphisms were found. These polymorphisms can serve as potential genetic markers. RAPD data were processed using cluster analysis and plotted as dendrogram. Each tested cultivar was clearly distinguished from the others. Moreover,Pisum sativum andP. sativum subsp.arvense cultivars were separated into 2 different clusters, according to their systematic relationships.     

利用RAPD标记研究荔枝品种的新缘关系

采用改进的ＣＴＡＢ方法提取３４个荔枝品种的基因组ＤＮＡ为模板,从５０个１０碱基的随机引物中获得了３７个多态性ＲＡＰＤ标记,聚类分析结果表明这３４个荔枝品种可以分成４组,第一组有“状元红”、“元红”、“桂林”、“宋家香”、“台湾荔”、“下番枝”、“东刘一号”、“妃子笑”、“白糖罂”、“香丸”、“绿沙”;第二组有“无栗子”、“水东”、“乌叶”、“蔡家肉丸”、“青壳糯米糍”、“糯米糍”;第三组有“桂味”     

Detection of DNA polymorphism among pea cultivars using RAPD technique

An influence of some Random Amplified Polymorphic DNA (RAPD) reaction factors on resulting banding pattern and the ability of RAPD technique to detect DNA polymorphism among six economically important pea cultivars was tested. Relatively high level of DNA polymorphism among peas was observed, using polyacrylamide/urea gels and silver staining. Altogether 13 arbitrarily designed primers produced 313 amplification products. In addition 59 polymorphisms were found. These polymorphisms can serve as potential genetic markers. RAPD data were processed using cluster analysis and plotted as dendrogram. Each tested cultivar was clearly distinguished from the others. Moreover,Pisum sativum andP. sativum subsp.arvense cultivars were separated into 2 different clusters, according to their systematic relationships.     

葡萄早熟芽变品种“早生高墨”的RAPD分析

以中国野生葡萄的部分株系、河岸葡萄、砧木品种S04和欧洲葡萄部分品种的幼叶为试材,采用改良CTAB法,提取葡萄基因组DNA的完整性好、纯度高。以所提葡萄基因组DNA为模板,共筛选随机寡核苷酸引物180个,对葡萄品种高墨及其早熟芽变品种早生高墨(紫玉)进行RAPD分析,结果发现引物OPW02和OPG06在两者之间扩增出了与葡萄早熟性状相关的多态性DNA片段OPW02—590、OPG06—1300和OPG06—400。这为进一步克隆、测序并分析它们的序列构成,了解葡萄早熟芽变的分子机理乃至其它果树植物芽变的分子机理提供了分子依据。     

24个菊芋品种（系）遗传多样性的ISSR标记分析

利用ISSR分子标记技术研究了来源于国内外不同区域的24个菊芋（Helianthus tuberosus Linn．）品种（系）的遗传多样性,并结合块茎特征采用聚类分析法探讨了它们的遗传关系。结果表明：用16条ISSR引物从24个品种（系）的基因组DNA共扩增出242条带,包含228条多态性条带,多态性条带百分率达94．2％,其中7条引物的多态性条带百分率达100．0％。各品种（系）间的遗传距离为0．19～1．01,遗传距离平均值为0．45。聚类分析结果显示：在遗传相似系数0．68处可将24个品种（系）划分为5类,第1类仅包含‘青芋1号’（‘Qingyu No．1’）,第Ⅱ类仅包含W12,第Ⅲ类包含W30、W42、‘青芋2号’（‘Qingyu No．2’）、W09、W18、W26和WSl,第Ⅳ类包含W06和W84,第V类包含‘青芋3号’（‘Qingyu No．3’）、w23、W36、W43、W54、W75、WS0、W62、W64、W79、S150、S138和W66;多数块茎特征相似的品种（系）被聚在一起,但也有部分块茎特征不同的品种（系）被聚在同一类中;部分品种（系）的聚类分析结果与其形态分类结果及地理分布不一致。研究结果表明：供试的菊芋品种（系）具有较高的遗传多样性,存在着较为频繁的基因交流;基于ISSR标记分析能较准确地揭示出菊芋品种（系）间的遗传多样性。     

龙眼果皮形态结构比较观察及其与果实耐贮运的关系

比较了福建省 1 0个主栽龙眼品种果实的果皮形态和结构 ,结果表明 :不同品种在果皮厚度、外果皮表面颜色、龟状纹、放射线、瘤状突、刺毛、外果皮皮孔、周皮层厚度、栓质层厚度和连续性、中果皮薄壁组织细胞排列、石细胞大小、含量、排列和分布 ,维管束发达状况、排列和分布 ,内果皮表皮细胞排列和角蜡质层厚度等方面均存在着明显差异。风梨味、东壁、油潭本、乌龙岭、红核子、蕉眼龙眼果皮厚 ,外果皮表面瘤状突和剌毛多 ,外果皮周皮层、栓质层厚且连续性好 ,中果皮石细胞 (团 )含量多且排列紧密 ,分布在中果皮外侧且在中果皮中所占比例大 ,维管束发达且排列有序 ,内果皮角蜡质层厚 ;这些品种果实耐贮运、抗病性强。而水涨、赤壳、福眼、普明庵龙眼果皮薄 ,外果皮周皮层薄、栓质层不发达 ,中果皮石细胞 (团 )含量少、分布分散 ,维管束不发达 ,薄壁组织细胞胞间隙大 ,皮孔间隙大、皮孔通道与中果皮组织细胞间隙相通 ;这些品种的果实不耐贮运、抗病性弱。讨论了龙眼外果皮表面主色为褐色和内果皮比外果皮更容易褐变的解剖学原因及龙眼果皮形态结构与果实耐贮运的关系。     

Direct comparison of levels of genetic variation in tomato detected by a GACA-containing microsatellite probe and by random amplified polymorphic DNA.

In this study, a direct comparison was made of the ability of four selected random amplified polymorphic DNA (RAPD) primers and a GACA-containing microsatellite probe to detect genetic variation in Lycopersicon. Of the 89 RAPD primers initially tested, 85 showed differences between a representative of Lycopersicon pennellii and L. esculentum, but only 4 distinguished among three L. esculentum cultivars. These four primers were subsequently tested on representatives of six Lycopersicon species. In pairwise comparisons of species, all or 14 of the 15 combinations could be distinguished by single primers. When the primers were tested on 15 L. esculentum cultivars, 90 of the 105 combinations could be distinguished by the four primers together. Finally, none of 118 tested primers showed reproducible differences among calli or progeny of régénérants from tissue culture, although some of the plants had inherited morphological mutations. The probe pWVA16, which detects GACA-containing microsatellites, could distinguish in TaqI-digested DNA the representatives of Lycopersicon species as well as all the L. esculentum cultivars tested. The probe was unable to detect polymorphisms among calli and the progeny of regenerants from tissue culture. An analysis of the results showed that the four selected RAPD primers were able to detect polymorphic bands among species at a frequency of 80%, and among cultivars at a frequency of 44%. In contrast, the microsatellite probe detected polymorphic bands at a frequency of 100 and 95%, respectively. The GACA-containing probe did not detect any common bands among the representatives of the six species, while band sharing with RAPDs was 48%. These results indicate that the two methods detect two types of DNA that differ in their degree of variability.     

Analysis of Genetic Diversity amongIxoraCultivars (Rubiaceae) using Random Amplified Polymorphic DNA

We have optimized the genomic DNA extraction method from freshand dry laminas, as well as fresh and dry corolla lobes ofIxoracultivars.Some woody tropical species such as these contain excessiveamounts of phenolic compounds that co-precipitate with DNA resultingin poor or no amplification during the polymerase chain reaction(PCR). Repeated precipitation with CsCl coupled with phenol:chloroformextraction yielded high quality DNA suitable for consistentPCR amplification. The DNA from fresh laminas of 22 cultivarsofIxorawas subjected to random amplified polymorphic DNA (RAPD)analysis. Individual taxa could be identified using specificDNA markers from the RAPD profiles. Cluster analysis of datafrom six primers grouped all 22 cultivars distinctly under twocultivar groups, viz.,IxoraCoccinea andI.Javanica. The percentagegenetic similarity was calculated for all the cultivars basedon the RAPD data. The two cultivar groups and the outgroup plantswere also clearly distinguishable with polar ordination usinga matrix of genetic dissimilarities (one minus similarity).Our data indicate that besides the use of RAPD markers for identificationof particularIxoracultivars within a germplasm collection, thephylogenetic relationships generated by RAPD analysis may beuseful for future breeding programmes. IxoraL. cultivars; Rubiaceae; RAPD fingerprinting; DNA extraction; woody tropical species     

利用RAPD研究桂林桂花品种间的亲缘关系

采用随机扩增多态性DNA(RAPD)技术,从100个随机引物中筛选出扩增效果较好的20个引物,分 析桂林市23个桂花品种的基因组多态性。20个随机引物共检测到193个位点,其中多态位点114个,占 59.1%。并进行了聚类分析,构建出树状聚类图,将这些品种划分为4个品种群,与传统分类学结果一致。结 果表明,以基因型而不是以表现型为基础,分析桂花品种间的区别是可能的。该技术为解决桂林市的桂花品 种分类问题提供了重要依据。     

玉米品种遗传多态性与根系内生细菌种群的相互关系

对辽宁省14个玉米主栽品种根系内主要细菌种群的分析结果表明,芽孢杆菌属（Bucillus ssp．）为玉米内生细菌的主要种群,其它种属包括肠杆菌属、沙雷氏杆菌属、假单孢菌属、黄单孢菌属和棍状杆菌属。内生细菌在不同玉米品种和不同生育期之间存在程度不同的差异。从90条随机引物中筛选出条带清晰、重复性好的13个引物,对14个玉米主栽品种进行遗传多态性的RAPD分析,共扩增出条带139条,其中多态性条带101条,多态性比率72．7％。利用SPSS统计分析软件对14个玉米品种在DNA水平和内生细菌种群水平上进行聚类分析,结果表明,品种亲缘关系在这两个水平上表现基本一致,即品种内生细菌的种类及其数量在很大程度上受品种的遗传背景控制。     

Estimation of Genetic Variability of Vigna radiata Cultivars by RAPD Analysis

DNA was isolated from 14 cultivars of Vigna radiata (L.) Wilczek and subjected to RAPD analysis using 14 random decamer primers. These cultivars revealed polymorphism with respect to RAPD markers and were subjected to hierarchical cluster analysis. A dendrogram was prepared based on these data. Analysis of banding patterns confirmed that two strongly aromatic cultivars IC1, IC4, were closely linked. But another aromatic cultivar, B1, formed a separate cluster. The high yielding cultivars were closely related to B1. The phylogenetic tree constructed by the neighbour joining method showed that RAPD results were correlated with morphological characters like plant height, leaf and seed size, seed colour, etc. This revised version was published online in July 2006 with corrections to the Cover Date.     

Identification and Assessing the Cultivars of Laminaria Lamx. （Phaeophyceae） with Molecular Markers

Molecular markers were used to identify and assess cultivars ofLaminaria Lamx. and to delineate their phylogenetic relationships. Random amplified polymorphic DNA (RAPD) analysis was used for detection. After screening, 11 primers were selected and they yielded 133 bands in all, of which approximately 99.2% were polymorphic. The genetic distances between gametophytes ranged from 0.412 to 0.956.Two clusters were formed with the unweighted pair group method with arithmetic mean (UPGMA) dendrogram based on the simple matching coefficient. All cultivars ofLaminariajaponica Aresch. used for breeding in China fell into one cluster. L.japonica from Japan, L. saccharina (L.) Lam., and L. angustata Kjellm.formed the other cluster and showed higher genetic variation than L. japonica from China. Nuclear ribosomal DNA (rDNA) sequences, including internal transcribed spacers (ITS1 and ITS2) were studied and aligned. The nucleotides of the sequences ranged from 634 to 668, with a total of 692 positions including ITS1, ITS2, and the 5.8S coding region. The phylogenetic tree obtained by the neighbor-joining method favored, to some extent, the results revealed by RAPD analysis. The present study indicates that RAPD and ITS analyses could be used to identify and assess Laminaria germplasm and to distinguish some species and, even intraspecies, in Laminaria.     

Identification and Assessing the Cultivars of Laminaria Lamx.(Phaeophyceae) with Molecular Markers

Molecular markers were used to identify and assess cultivars ofLaminaria Lamx. and to delineate their phylogenetic relationships. Random amplified polymorphic DNA (RAPD) analysis was used for detection. After screening, 11 primers were selected and they yielded 133 bands in all, of which approximately 99.2% were polymorphic. The genetic distances between gametophytes ranged from 0.412 to 0.956.Two clusters were formed with the unweighted pair group method with arithmetic mean (UPGMA) dendrogram based on the simple matching coefficient. All cultivars of Laminaria japonica Aresch. used for breed ing in China fell into one cluster. L. japonica from Japan, L. saccharina (L.) Lam., and L. angustata Kjellm.formed the other cluster and showed higher genetic variation than L. japonica from China. Nuclear ribosomal DNA (rDNA) sequences, including internal transcribed spacers (ITS1 and ITS2) were studied and aligned. The nucleotides of the sequences ranged from 634 to 668, with a total of 692 positions including ITS1, ITS2, and the 5.8S coding region. The phylogenetic tree obtained by the neighbor-joining method favored, to some extent, the results revealed by RAPD analysis. The present study indicates that RAPD and ITS analyses could be used to identify and assess Laminaria germplasm and to distinguish some species and, even intraspecies, in Laminaria.     

RAPD分析花椰花不同品种的遗传变异

利用随机扩增多态性DNA（RAPD）方法,对13个花椰花品种的基因组DNA进行多态性分析。选用20个10bp随机引物,共扩增出175条DNA片段,其中多态性片段118例,占67．4％,结果表明,花椰花品种间具有丰富的遗传多样性。依据扩增结果进行遗传相似系数分析,构建聚类分析树状图。初步探讨了各品种间的遗传变异关系及RAPD技术在花椰花种质资源分类鉴定和育种工作中的应用前景。     

青岛市19个樱花品种的RAPD分析

采用改良的CTAB法提取19个樱花品种的基因组DNA.利用RAPD技术对19个樱花品种进行亲缘关系鉴定和品种分类研究.从60条10 bp随机引物中筛选出32条扩增效果较好的引物进行扩增,共扩增出533条带,其中多态性带为406条,多态率达76.17%.根据扩增结果进行UPGMA聚类分析,聚类结果将19个品种分为2大类群,第1类群主要为杂交樱系;第2类群为日本晚樱系,根据樱花的重瓣性、枝姿和花色又可分为不同的亚类群、类和亚类.结果表明应用RAPD技术对樱花分子水平的分类结果与传统分类学的结果基本一致,进一步证明种源、重瓣性、枝姿和花色都可作为樱花品种分类的重要指标.