十二份杧果种质亲缘关系的SRAP分析

以12份杧果种质为试材,利用SRAP分子标记技术对其进行遗传多样性研究。结果表明:从36对引物中筛选出22对多态性好的引物,共扩增出251条DNA条带,其中多态性谱带181条,平均每对引物扩增得到15.08条多态性谱带,多态性比率为72.11%。UPGMA聚类表明,所有供试品种(系)之间的亲缘关系都比较近,相似系数在0.77~0.89;如果以相似系数0.78为标准,可将供试的12个品种(系)分为3类。表明利用SRAP技术能较好的区分杧果的亲缘关系;SRAP分子标记适合杧果的DNA遗传多样性分析,可作为杧果种质鉴定依据之一。     

紫色叶用莴苣遗传多样性及亲缘关系的TRAP分析

利用TRAP标记对47份紫色叶用莴苣种质进行亲缘关系及遗传多样性分析。20对引物组合共扩增出多态性条带430条,多态性比率(PPB)为67.08%。47份紫色叶用莴苣材料的遗传相似系数在0.7063~0.9828之间,Nei’s基因多样性指数(He)和Shannon’s信息指数(I)分别为0.2553和0.3976,遗传多样性水平较低。聚类分析结果表明,47份紫色叶用莴苣材料分为2大类群,第Ⅱ类群又分为5个亚类,叶片形态相似的种质基本上聚在一起,亲缘关系较近。     

杏92份种质资源的ISSR分析

利用ISSR分子标记对92份杏抗寒种质资源进行了遗传多样性分析。从35条引物中筛选出10条用于杏资源的ISSR扩增,共扩增出86条带,其中多态性条带83条,多态性百分比为96.5%。用UPGMA法聚类分析,结果显示,龙垦14号杏(Armeniaca vulgaris)、A1B20(A.vulgaris)、牡红杏(A.vulgaris)3个普通杏品种聚为1类,辽杏(A.mandshurica)单独聚为1类,其他88份杏资源聚为1类,不同种质资源间表现出丰富的遗传多样性。     

安徽及周边地区枣种质资源遗传多样性研究

利用ISSR标记对32份安徽省地方枣种质资源及山东、河南等邻近省份的42份枣资源进行了遗传多样性分析。26条引物在74份种质中共扩增出241个位点,其中多态性位点214个,多态性比率为88.8%。利用NTSYS软件计算得到DICE相似系数为0.5936 ~ 0.9353;32份安徽地方种质共扩增出219条电泳谱带,其中186条为多态性条带,多态性比率为84.93%,32份样品遗传相似系数在0.6267 ~ 0.9353之间,表明安徽地方枣种质资源遗传多样性较为丰富。采用UPGMA法对74份种质的ISSR数据进行聚类分析,分析结果显示可分为8个类群,种质间亲缘关系与地理来源关系较为密切,来自安徽的地方种质基本上都独立聚成不同的类群,表明安徽地方枣种质与周边其它省份枣种质资源亲缘关系较远;研究结果表明体细胞突变可能是推动安徽地方枣种质进化的重要因素。     

枸杞种质遗传多样性SCoT分析

以17份枸杞种质为试材,采用SCoT标记法,分析了17份枸杞种质资源的遗传多样性,以期为枸杞种质资源的深度开发与利用提供参考。结果表明:从42条引物中筛选出7条重复性好、条带清晰的引物进行PCR扩增,共扩增出32条条带,其中30条条带具有多态性,多态性比率为93.47%;UPGMA聚类分析显示,17份枸杞种质间遗传相似系数在0.30~1.00,平均相似系数为0.80,在遗传相似系数为0.72时,可将17份枸杞资源分成四大类。表明采用SCoT标记法能够揭示枸杞种质间的遗传多样性。     

不同产地人参种质资源RAPD和SSR分析

以15个产地人参种质资源为试材,采用RAPD和SSR分子标记技术对其遗传多样性进行分析。结果表明:2种分子标记均能揭示不同地区人参种质间的遗传多样性。共筛选出11条RAPD随机引物,平均每条引物可扩增出3~17条DNA片段,共扩增出104条清晰条带,多态性条带100条,多态性百分率为96.15%,揭示供试材料间遗传相似系数(GS)为0.505 3~0.989 5,平均值为0.760 4。筛选出5对SSR引物,平均每对引物可扩增出1~8条DNA片段,共扩增出38条清晰条带,多态性条带35条,多态性百分率为92.11%,揭示供试材料间遗传相似系数(GS)为0.400 0~0.960 0,平均值为0.742 1。RAPD和SSR标记的聚类分析在分类上稍有差异,但总体趋势一致,RAPD、SSR及RAPD+SSR均将样品聚为三大类,均能揭示它们之间的亲缘关系,为人参种质资源的收集与利用奠定了基础。     

菠萝遗传多样性的SRAP分析

利用SRAP标记研究了61份菠萝种质的遗传多样性。从99对SRAP引物中筛选出35对引物对61份菠萝种质进行遗传多样性分析,均得到了清晰指纹,且所有品种表现出多态性;供试菠萝品种的多态性比率均较小,在47.32%~68.90%;35对SRAP标记共获得567条带,多态性位点384个,多态条带比率为67.72%;61份菠萝种质其相互之间相似系数为0.8027~0.9510,表明61份菠萝品种之间遗传关系相对较近;应用UPGMA进行聚类,在大致相似系数0.8330的水平上,61份材料可分为5种类型。     

萱草部分野生种和栽培品种亲缘关系的AFLP分析

 借助AFLP标记对35份萱草野生种和栽培品种进行亲缘关系研究, 结果表明, 7对引物组合对萱草共扩增出条带380条, 其中多态性条带357条, 平均多态性达到93.39% , 单对引物扩增条带19～85条, 平均每对引物组合扩增多态性条带51条。7对引物扩增出的多态性条带均超过90.0%。种质资源相似系数为0.3822～0.9656, 平均相似系数为0.7039。UPGMA聚类结果将供试材料分为3类, 即早花、中花和晚花类, 同一产地的品种基本能聚在一起。AFLP标记技术能较好地从分子水平揭示萱草种质资源的亲缘关系。     

菠萝蜜遗传多样性的ISSR分析

用ISSR标记方法对76份菠萝蜜(Artocarpus heterophyllus Lam.)种质资源DNA的遗传多样性进行了检测,24个引物共检测到477条带,其中427条具多态性(占89.52%),平均PIC为0.23,24个引物能把76份种质完全区分开来。遗传距离分析结果显示供试种质的遗传多样性较低,在DNA水平上的遗传相似系数为0.626～0.945,平均为0.775。聚类分析表明,76份菠萝蜜材料在遗传距离系数为0.752处可分为4大类,其中热带植物园的种质WJ2和GSYWJ1与其他种质的遗传距离相对较大,干胞和湿类型不能独立聚类。另外各地区的种质与雷州半岛种质明显分开聚类,而雷州半岛各地区的种质混杂聚类在一起,不能按地区单独聚类。     

西葫芦种质资源遗传多样性的RAPD分析

利用RAPD技术对国内外47份西葫芦种质材料进行遗传多样性及亲缘关系分析。从520条随机引物中筛选出30条能产生稳定多态性的引物用于RAPD分析,共扩增出367条带,其中302条表现多态性,占总数的82.29%,表明西葫芦种质资源在分子水平上具有丰富的遗传多样性,与形态学研究结果一致。聚类分析将47份西葫芦种质材料分为两大类。在第Ⅰ类群中,可将32份种质分为4个亚类;第Ⅱ类群分成5个亚类。从生长习性来看,矮生与半蔓生的亲缘关系最近;从皮色来看,白色与绿色亲缘关系最近,亲缘关系最远的为白色与花皮。     

Genetic relatedness (diversity) and cultivar identification by randomly amplified polymorphic DNA (RAPD) markers in teasle gourd (Momordica dioica Roxb.)

Genetic relationship and variation of 29 accessions of teasle gourd (Momordica dioica Roxb.) and 1 accession of Momordica cochinchinensis Spreng. (wild relatives of teasle gourd) were examined by RAPD analysis using 44 dodecamer oligonucleotide primers. A total of 496 fragments were produced by 44 primers of which 95% bands were polymorphic. Using presence or absence of specific RAPD markers or combination of primers, 23 out of 30 accessions were identified. The genetic relatedness or genetic distance based on Nei and Li's genetic similarity varied from 0.86 to 0.65 with an average of 0.74 among 29 M. dioica accessions (when M. cochinchinensis excluded). In the phenetic dendrogram developed from cluster analysis using UPGMA method, M. cochinchinensis was out grouped as single accession, while others showing relatively weak grouping formed four groups. Clustering pattern did not demonstrate any relationship between geographical origin and genetic diversity. A DNA extraction method has been standardized. This is the first report of using RAPD techniques in teasle gourd. It was concluded that RAPD analysis is a useful tool for genotypic identification and estimation of genetic similarity in teasle gourd.     

Comparative analysis of genetic diversity in Indian bitter gourd (Momordica charantia L.) using RAPD and ISSR markers for developing crop improvement strategies

A genetic analysis of 38 diverse Indian bitter gourd (Momordicacharantia var. charantia, and var. muricata) accessions was performed using 29 RAPD and 15 ISSR markers. RAPD primers yielded 208 amplicons of which 76 (36.5%) were polymorphic providing an average of 2.6 amplicons per primer. RAPD amplicons per primer ranged from 3 (OPE-19, OPW-09) to 15 (OPW-05), and varied in size from 200 bp to 3000 bp. Fifteen ISSR primers provided a total of 125 bands of which 94 (74.7%) were polymorphic. Polymorphic ISSR markers ranged from 0 (UBC-841) to 12 (UBC-890) providing a mean of 6.3 amplicons per primer that ranged in size from 150 bp to 2700 bp. Nevertheless, the concordance among bitter gourd accession groupings after cluster analysis was relatively high (r = 0.77), indicating that RAPD- and ISSR-based diversity assessments in this germplasm array were generally consistent. The M.charantia var. charantia (domesticated) and var. muricata (wild, free-living) accessions examined were genetically distinct, and these differences provided for the development of strategies for genetic analyses and crop improvement in this species.     

Assessment of genetic diversity among mango (Mangifera indica L.) genotypes using RAPD markers

Knowledge about the extent of genetic diversity/relatedness in mango germplasm is vital for developing coherent strategies for future gains in productivity. The genetic diversity/relatedness among mango cultivars/genotypes developed in Pakistan has not been investigated previously. We have assessed the genetic diversity among 25 mango genotypes/cultivars using randomly amplified polymorphic DNA (RAPD). Sixty random ten-mer primers were surveyed, out of which 45 yielded amplicons in all the genotypes. Genetic similarity between genotypes/cultivars was in the range of 64–89% with an average of 74%. Similarly, the genetic relatedness among all variants derived from a mango cultivar Chaunsa was in the range of 81.18–88.63%. These coefficients were utilized to construct a dendrogram using the unweighted pair group of arithmetic means (UPGMA). The genotypes were grouped into three (A, B, C) clusters. Generally, genotypes originating from Pakistan were grouped in cluster ‘A’ while cluster ‘B’ primarily composed of southern India as well as Florida cultivars. Kensington Pride was the most distantly related genotype which grouped with Maya and Yakta, forming a distinct cluster ‘C’.     

Characterization of sequence-related amplified polymorphism markers analysis of tree peony bud sports

Bud sports have been widely exploited for creating novel cultivars in vegetatively propagated plants. However, bud sports of tree peony have rarely occurred nor been applied to breeding. We used sequence related amplified polymorphism (SRAP) markers to characterize two tree peony groups with bud sports. ‘Er Qiao’ and ‘Luo Yang Hong’ formed group I and are derived from Chinese cultivars, while ‘Shima-nishiki’ and ‘Taiyō’ formed group II and are related to Japanese cultivars. The DNA samples were evaluated with 25 different primer pairs. This resulted in a total of 149 different amplicons in Group I, of which 80.5% were polymorphic. Group II generated a total of 57 amplicons, of which 63.2% were polymorphic. The two groups shared 43 amplicons. The genetic similarity among the cultivars ranged from 0.33 to 0.60. The two bud sports groups were apparently separated on the unweighted pair-group method with arithmetic averages cluster analysis tree. The results demonstrated that this marker system appeared effective in detecting molecular differences between original parents and their bud sports in tree peony. This study provides a technical base for characterizing plants with bud sports, which are valuable genetic resources for further breeding.     

辐射诱变和芽变柑橘品种(系)的AFLP分析

为了对广东省近年来新选育的16个60Co-γ辐射诱变和芽变柑橘优良品系(株系)进行遗传差异分析,应用AFLP技术,对3个亲本和16个辐射诱变和芽变的柑橘品种(系)进行了遗传差异分析。从64对引物中筛选出9对多态性高、分辨能力强的引物进行分析,结果表明:(1)与各自的亲本相比,供试16个辐射诱变和芽变的柑橘变异类型的遗传基础均已经发生了不同程度的变化。(2)利用引物EcoRⅠAAG+MseⅠCTT绘制供试样品的AFLP指纹图谱,并根据各自的差异带或特征带区分了19个柑橘试材。(3)对AFLP扩增结果进行聚类分析,根据相似系数0.77的水平可将19个样品分为4组。红江橙品系中的华青1号与亲本间的相似性系数仅为0.647 7,可将其单独划为一组。可为柑橘新品种选育研究的早期鉴定提供参考依据。     

Genetic diversity of Chinese natural bermudagrass (Cynodon dactylon) germplasm using ISSR markers

Bermudagrass (Cynodon dactylon L.) is an important warm-season turfgrass. Although it is widely distributed in China, studies on the genetic variation and relationship among the large-scale indigenous bermudagrass were relatively insufficient, especially at molecular level. The purpose of this study was to assess the molecular variation and relationship among one commercial cultivar ‘Tift3’ and 95 wild bermudagrass accessions collected from 11 provinces in China by ISSR marker technique. The results indicated that 29 ISSR primers generated a total of 248 bands among which 242 (97.6%) were polymorphic bands. The genetic similarity coefficients among accessions ranged from 0.51 to 0.97 with an average of 0.74. All accessions could be clustered into 11 groups with UPGMA method. Accessions from the same or adjacent regions generally were clustered into the same group or subgroups. A few accessions, however, were greatly different from the majority of all accessions. The results suggested that ISSR marker is an effective tool for the study of genetic variation in Chinese natural bermudagrass.     

24个大果沙棘品种的RAPD分析

为了解沙棘种质资源的遗传多样性和品种间的亲缘关系,以24个大果沙棘品种为材料,利用随机扩增多态性DNA(RAPD)方法进行沙棘的遗传多样性分析。从120个随机引物中筛选出多态性丰富的18个引物用于扩增反应,在24个大果沙棘品种中检测到171个RAPD位点,其中多态性DNA带112条,多态位点百分比为65.5%,表明大果沙棘品种具有丰富的遗传多样性;在RAPD分析的基础上,对大果沙棘品种进行了聚类分析,结果表明,以相似系数0.58处作为分界,可将供试的24份大果沙棘种质分为4类。     

Study of genetic diversity in Chamomile (Matricaria chamomilla) based on morphological traits and molecular markers

Chamomile is one of the most important medicinal plants in the world trade that has many applications in drug and sanitary industrials. In order to evaluate the genetic diversity of different chamomile landraces based on morphological and molecular markers, 20 landraces were collected from different area of Iran. In addition to that, five populations imported from European were examined. The augmented design with four blocks and five controls were used to assess morphological traits. The RAPD method was utilized for evaluating the genetic diversity. Results showed that the economical yield, the number of flowers in plant, and the essential oil content had maximum variance coefficient. The flower's diameter and height had minimum variance coefficient. According to the cluster analysis on both morphological and molecular markers, 25 populations were classified into 5 clusters, but the population intra-groups were different. From 29 reliable primers that were used, 369 bands were detected and from which 314 (85.44%) bands were polymorphic. Genetic Jaccard's similarity coefficient was estimated in the range of 0.15–0.63, and with a mean of 0.35. Results showed that the genetic diversity was not according to the geographical diversity.     

Genetic similarities among cocoyam cultivars based on randomly amplified polymorphic DNA (RAPD) analysis

Eighteen cultivars of cocoyam (Xanthosoma spp.) and two cultivars of taro (Colocasia esculenta (L.) Schott) from the USDA/ARS germplasm collection were evaluated for genetic relatedness using RAPD data. Seven random primers generated 40 RAPD loci. Of the 18 cultivars screened, 11 (61%) were identical at all RAPD loci evaluated. A similarity matrix was constructed on the basis of the presence or absence of bands. Among cocoyam cultivars the genetic similarity ranged from 0.86 to 0.97 with a mean of 0.91. Cluster analysis identified two main clusters with some unexpected groupings. These data indicate that very little genetic variation exists within the accessions used in this study and that this Xanthosoma spp. collection is of limited value as a genetic resource.     

利用RAPD技术评估金针菇种质资源

本文利用20个10核苷酸的随机引物对来源不同而具有代表性的16个金针菇菌株进行了RAPD分析研究。结果表明:所测试的20个随机引物中有6个引物的扩增产物DNA片断表现出明显的多态性,对所有供测试的金针菇菌株总共扩增出84条具重复性的DNA片断;同时也显示供试的16个金针菇菌株两两间的遗传相似性变化较大(相似系数从0.0592到0.8000)。另外,采用系统聚类法中的类平均法,对供试的16个菌株两两间的相似系数进行聚类,可将它们分为四大类,各大类的类间和类内菌株的遗传变异程度较大,其中以Ⅳ类内各菌株间的最高(平均相似系数为0.4353),Ⅰ类和Ⅱ类间各菌株间的最低(平均相似系数为0.6812)。但是,从整个被测试的16个金针菇菌株来看,它们之间的遗传变异并不丰富(总平均相似系数为0.5292)。同时,将RAPD技术应用于不同菌株间遗传差异的研究,具有反应迅速、不受外界环境条件影响、能从DNA分子水平上揭示菌株间的遗传差异等优点。因此,RAPD分析技术是一种快速准确评估食用菌种质资源的有效方法。