中国马铃薯部分栽培品种遗传多样性的AFLP分析

 采用AFLP方法分析中国各地79个马铃薯栽培品种, 获得了8对引物组合的扩增谱带, 为马铃薯品种的鉴定提供了分子依据。8对引物组合扩增结果共获得801条带, 平均每对引物组合产生100.13条带。其中493条为多态性条带, 平均多态性检出率为61.2% , 引物组合E11 /M51扩增的多态性比率最高。聚类分析将材料分成3类, 聚类结果与地域无明显相关。     

Genetic diversity in fruiting-mei,apricot, plum and peach revealed by AFLP analysis

Summary

High-throughput amplified fragment length polymorphism (AFLP) analysis, employing a two-dye automated DNA sequencer, was used to genotype accessions of four important Prunus spp. fruit crops in China, fruiting-mei (P. mume), apricot (P. armeniaca), plum (P. salicina, and P. ussuriensis) and peach (P. persica). Nine primer pairs generated a total of 2,089 AFLP bands, of which 94.4% were polymorphic. Levels of polymorphism and the genetic relationships among the crops were studied.The fixation index (F_st) of the four different fruit trees ranged from 0.53 – 0.82, and the average genetic diversity (AGD) in these species ranged from 3.2% to 12.8%. These four stone fruit species could be clustered into four clear groups on a phylogenetic tree based on bootstrap analysis. Phylogenetically, fruiting-mei was shown to be closest to apricot and most distant from peach.     

苦瓜种质资源ISSR遗传多态性分析

利用ISSR分子标记对30份苦瓜种质进行遗传多态性分析,从23个引物中筛选出13条重复性好,条带清晰的引物进行PCR扩增,共扩增出79条带,其中58个为多态性条带,占总带数的73.4%,每个引物扩增的条带数为4~10条,平均6.07条。对ISSR结果进行聚类分析,结果表明,30份苦瓜种质间的遗传距离为0.025~0.520。在遗传距离为0.21处可将30份苦瓜种质划分成6个品种群,其亲缘关系与种质地理分布和植物学性状特征等有一定关联。     

Analysis of genetic diversity in Portuguese Ceratonia siliqua L. cultivars using RAPD and AFLP markers

Although carob (Ceratonia siliqua L.) is of great economic importance little is still known about the pattern of genetic variation within this species. Morphological characteristics based on 31 fruit and seeds of continuous characters determinant for agro-industrial uses, were compared with RAPD and AFLP markers for assessing genetic distances in 68 accessions of carob trees, from different cultivars, varieties and eco-geographic regions of Algarve. Eighteen selected RAPD primers applied to the 68 accessions produced a total of 235 fragments ranging from 200 to 2000 bp, of which 93 (40%) were polymorphic. Four AFLP selective primer combinations generated a total of 346 amplification fragments of which 110 were polymorphic. The average level of polymorphism based on four primer combinations was 31.8%. The phenetic trees based on RAPD and AFLP analyses gave high co-phenetic correlation values, and were found to be consistent in general with the analysis of morphological data, carried out on the same accessions. A number of RAPD and AFLP markers were found to be diagnostic for ‘Canela’ cultivar and 13 wild ungrafted trees.     

Detection of genetic diversity among populations of sweet cherry (Prunus avium L.) by AFLPs

Summary

The high degree of polymorphism of AFLPs provides an efficient system for identification and genome analysis of sweet cherry (Prunus avium) cultivars and selections. The cultivars of sweet cherry have usually been characterized by assessment of phenotypic and pomological traits. AFLP markers were employed to identify 38 sweet cherry accessions and estimate the genetic diversity among this material. Ten of 18 tested primer combinations were informative with up to 80 bands per primer combination. Seven to 33% of the amplfied bands were polymorphic depending upon primer combination. Allcultivars and selections tested could be clearly identified. The objective of this work was to demonstrate the usefulness of molecular markers in revealing the genetic diversity among different sweet cherry genotypes.     

Genetic characterization of asparagus doubled haploids collection and wild relatives

The genus Asparagus is very large consisting of around 150 species found as herbaceous perennials, tender woody shrubs and vines. The cultivated species (Asparagus officinalis L., diploid) is a highly prized vegetable, grown in different environments ranging from cool temperate zones to deserts, Mediterranean climates and tropical areas. As a consequence, Asparagus breeders have developed different cultivars that differ for their morpho-agronomic traits, habit and ploidic status (few triploid and tetraploid cultivars are used). Several breeding methods are used for developing cultivars, among which a well developed in vitro anther culture technique produces homozygous clones useful for F₁ hybrids constitution. A fluorescent based AFLP (amplified fragment length polymorphism) technique were applied with the aim to assess genetic diversity among a collection of 173 doubled haploid (DH) androgenetic clones, five Asparagus wild species and interspecific hybrids obtained among the cultivated species and two wild relatives. The average number of AFLP fragments generated per primer set was 105, varying in size from 50 to 550 bp. A total of 1054 AFLP fragments were detected, 20% of which were polymorphic. Genetic similarity based on DNA polymorphisms, showed that a few number of AFLP primer combinations are able to distinguish the cultivated DH clones from the wild species. Indeed, from one DH clone for each anther donors and the wild species were used to construct a dendrogram using Dice's coefficient and the unweighted pair group method with the arithmetic mean (UPGMA). Genetic distances among all DH clones were calculated using the C.S. Chord distance; and a neighbour-joining (NJ) consensus tree was constructed in order to support the breeder for parental genotype choice for asparagus hybrid constitution.     

Identification of 57 sweet orange cultivars using AFLP markers

Summary

Sweet orange (Citrus sinensis) represents an important group of Citrus fruit; however, the identification of sweet orange cultivars during vegetative growth can be difficult. A study on the genetic identification of sweet orange cultivars may be significant for the sweet orange nursery industry, for cultivar-rights protection, and is important for the genetic evaluation and conservation of these orange cultivars. In this study, amplified fragment length polymorphism (AFLP) markers were used to genotype 57 sweet orange cultivars. Ten PCR primer pairs generated 629 unique AFLP bands, with a size range of 50 ? 500 bp. Seventy-four bands (11.8%) were polymorphic. On average, each primer pair produced 62.9 fragments, with 7.4 polymorphic fragments. A dendrogram of the 57 sweet orange cultivars was constructed based on an UPGMA analysis using Jaccard?s coefficients of similarity. This provided a clear comparison of the genetic variation between cultivars and an ability to identify them. From Jaccard?s coefficients, 56 of the 57 cultivars examined were genetically close, with coefficient values ≥ 0.985. ?Variegated Navel? was less closely-related, with a much lower coefficient value (0.94). Among the 57 cultivars, 28 sub-groups, some consisting of only one cultivar, could be separated by their AFLP fingerprints. Compared to ISSR and SSR markers, AFLP seemed to be the preferential marker technique for the identification of sweet orange cultivars.     

Evaluation of genetic relationships among Iranian pistachios using microsatellite markers developed from Pistacia khinjuk Stocks

To evaluate the genetic relationships among wild and cultivated Pistacia species grown in Iran and the analysis of genetic variation among Iranian pistachio genotypes, two DNA libraries enriched for dinucleotide (AG)_n and trinucleotide (ATG)_n microsatellite motifs were developed from Pistacia khinjuk genome. Following screening of clones by colony PCR technique, 44 clones were sequenced and 27 pairs of primers designed from flanking regions of the repeats. The examination of primer pairs, designed from P. khinjuk sequences, showed successful cross-species amplification within the genus Pistacia. A dendrogram constructed on the basis of the Minimum Evolution clustering algorithm revealed that Pistacia vera has closer relationships with P. khinjuk, than with Pistacia integerrima, Pistacia palaestina, Pistacia atlantica and Pistacia mutica. The dendrogram further distinguished the wild Sarakhs pistachio from the rest of P. vera genotypes suggesting that the domesticated genotypes of P. vera are evolved from P. vera var. Sarakhs and then this wild genotype likely develops to other local pistachios. Hence, it seems that the wild Sarakhs pistachio plays an important role in evolutionary trend of the edible pistachios in Iran. The results indicated that microsatellites developed in P. khinjuk are distributed in the genome of indigenous pistachio species including P. vera genotypes and therefore they will be useful in characterization of Iranian pistachio genotypes.     

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。     

云南芋种质资源遗传多样性的AFLP分析

 采用荧光标记引物的AFLP分子标记技术, 用筛选出的“3 + 2”引物组合, 对48份云南芋种进行遗传多样性分析, 3对引物共扩增出184个DNA位点, 平均每对引物可检测出56.3个多态性位点,多态性位点高达91.8% , 云南芋种质资源在DNA分子水平上表现出极为丰富的遗传多样性。聚类分析表明: 野生种质和栽培种质的亲缘关系较远, 栽培种质基于AFLP标记的分类结果与形态性状基本一致, 少数材料差异较大。     

Development of sex-associated RAPD markers in wild Pistacia species

Summary

Marker assisted selection may greatly facilitate pistachio rootstock breeding as well as cultivar improvement, because of the long juvenile period of Pistacia species. Early diagnosis of seedling sex type would assist breeding and nurserymanagement in these dioecious species. We searched for RAPD markers linked to sex in P. atlantica, P. terebinthus and P. eurycarpa, the main wild species in Turkey that are used as rootstocks for P. vera. For this purpose, leaf samples were collected from male and female individual trees from each species and sex-pooled DNA samples were prepared by mixing the DNA of ten male and ten female individuals, to screen for sex associated RAPD bands. A total of 472 primers have been screened so far and two bands, amplified by primers BC156 and BC360, appeared to be sex assocaited in P. eurycarpa. The bands were tested in 30 male and 37 female individuals. Band BC156 (1300) was present in all, except one, female trees and was absent from all the male trees. Band BC360 (500) was amplified in 31 out of 37 females and was absent from all the males. In P. atlantica, one primer, OPAK09, amplified a femaleassociated band (850 bp), that was present in all 46 female individuals tested and absent in all the 38 male trees tested. It is likely that these markers are linked to sex-determining loci. The sex determination mechanism has not been characterized in Pistacia and segregating populations from controlled crosses are required to elucidate such mechanism and also to measure the genetic distance of our markers from the putative sex loci.     

山楂（Crataegus pinnatifida Bge.)遗传多样性的RAPD和ISSR标记分析

 利用RAPD和ISSR标记对35份山楂（Crataegus pinnatifida Bge.）资源进行了DNA多态性分析。12个RAPD引物共扩增出110条清晰的谱带,其中89条显示多态性,平均每个引物扩增出7.4条多态性谱带。13个ISSR引物共扩增出110条清晰的谱带,其中94条显示多态性,平均每个引物扩增出7.2条多态性谱带。基于RAPD和ISSR标记,利用UPGMA分别构建了35份山楂资源的聚类树状图。距离系数分别为0～0.62（RAPD）和0～0.64（ISSR）,表明山楂具有较高的遗传多样性。     

青菜品种亲缘关系AFLP分析

利用AFLP技术对市场上主栽的10个青菜主要品种进行了遗传多样性及聚类分析.结果表明:最终筛选出的5对引物共扩增出多态性位点454个,多态性位点占总扩增位点的比例平均为57.5%,系统聚类分析将供试材料分为4组,基于分子标记的分类与材料的表现基本吻合,为以后品种的鉴定和分子辅助育种提供一定的参考.     

Genetic relationships of gladiolus cultivars inferred from fluorescence based AFLP markers

Gladiolus is one of the important commercial flowers with a large number of cultivars. However, genetic relationships among its genotypes have not been reported. This study analyzed genetic relatedness of 54 gladiolus cultivars using amplified fragment length polymorphism (AFLP) markers. A total of 24 AFLP primer pairs with three samples were initially screened, from which 9 primer sets that showed clear scorable and highly polymorphic bands were selected for AFLP reactions. Fluorescence-labeled amplification products were subjected to electrophoresis and then analyzed using an automated sequencer. A dendrogram was constructed by the unweighted pair group method using the arithmetic average (UPGMA). The number of AFLP fragments generated per primer set ranged from 10 to 151 with fragment sizes varying from 50 to 450 bp. A total of 660 AFLP fragments were detected, of which 658 (99.70%) were polymorphic. All the primers except E-AGG/M-CTA displayed 100% polymorphism. All cultivars were clearly differentiated by their AFLP profiles. The AFLP data were compared with previously obtained RAPD data and combined to generate a common dendrogram. The first cluster was dominated with indigenously bred cultivars while the second was dominated with exotic cultivars. This shows that most of the exotic cultivars as well as indigenous cultivars are closely related with each other. However, two indigenous cultivars viz., Pusa Suhagin and Pusa Archana share genetic similarity with exotic cultivars. Among the genotypes selected for the investigation, Pusa Gunjan was identified as the most distinct genotype. The AFLP markers developed will help future Gladiolus cultivar identification, germplasm conservation and new cultivar development. The assessed genetic relationships among gladiolus cultivars may enhance the efficiency of breeding program by selecting desirable parents with reduced breeding cycle.     

Assessment of Genetic Variability in Pistachio (<Emphasis Type="Italic">Pistacia vera</Emphasis> L.) with Nuclear SSR Molecular Markers

Pistachio (Pistacia vera L.) is considered to be the most widely cultivated species in the genus Pistacia and regarded as the most probable ancestral species within the genus. As one of the oldest nut crops in human history, pistachio nuts have a high nutritional value and are commercially important. In the present study, the genetic variation of pistachio genotypes was investigated by nuclear SSR markers. The transferability of the SSR markers was high across genotypes, thus, flanking regions of the SSRs are conserved in the studied genotypes. High level of polymorphism was detected among the studied genotypes. The seven SSR primer pairs generated a total of 18 alleles that 13 of them were polymorphic among the genotypes. The range of the polymorphic alleles was 1 (for Ptms9, Ptms40, Ptms41, and Ptms42) to 5 (for Ptms7 locus) with an average of 2.57. The amplified allele sizes ranged from 120 to 250?bp. Pair-wise genetic similarity coefficients varied from 0.20 to 0.75. The UPGMA dendrogram differentiated the genotypes into two major clusters. The present results may be used for the conservation, core collection and future breeding of the pistachio.     

梨新品种及其亲本的AFLP分析

利用荧光AFLP技术, 对20个梨新品种及其23个亲本(共43个品种) 进行研究。从64对引物组合中筛选出7对用于扩增, 共获得784条带, 其中多态性带699条, 多态性为89.2%。扩增结果显示, 7对引物组合在29个品种中扩增出特征带, 每对引物组合均能将所有品种鉴别开, 表明荧光AFLP技术用于梨品种鉴定的效率很高。通过聚类, 从分子水平对梨新品种及其亲本的遗传关系进行分析, 并对20个梨新品种进行分类, 为梨杂交育种的亲本选配提供了理论参考。     

红肉猕猴桃种质资源果实性状及AFLP 遗传多样性分析

 对中国红肉猕猴桃种质资源进行收集和调查,并对其进行果实性状变异分析和AFLP 遗传多 样性及遗传关系分析。结果表明,红肉猕猴桃野生资源主要分布于湖南省、湖北省、河南省、江西省、 四川省和陕西省等地,共采集到52 份野生资源和2 份品种资源（包括软枣猕猴桃红肉类型、中华猕猴桃 红肉类型和美味猕猴桃红肉类型）。红肉猕猴桃种质资源在果实性状和DNA 分子水平上都存在丰富的变 异和较高的遗传多样性水平,4 对AFLP 引物共扩增出259 个多态性位点,多态性位点百分率为90.56%, Nei’s 基因多样性和Shannon’s 信息指数分别为0.318 和0.477;资源间遗传相似性系数介于0.568 ~ 0.883 之间,平均为0.714。聚类分析和主坐标分析将54 份资源划分为4 个组,软枣猕猴桃红肉类型单独聚为 一类;中华猕猴桃和美味猕猴桃红肉类型亲缘关系较近且有按地理来源优先聚类的趋势。果实性状数据 和AFLP 数据之间具有极显著的相关性,二者可结合用于红肉猕猴桃资源评价和保护利用工作中。     

枳属36份特异种质的AFLP指纹图谱构建与分析

 从40个Pstl／MseI酶切的AFLP引物对中筛选出多态性高的7个，对36份枳属特异种质进行了指纹分析。7个引物对共获得539条带，其中146条带具有多态性。各种质材料之间的遗传相似性在0．23～ 0．98之间，只需结合其中的3个引物对即可区分所有的材料。根据7个AFLP引物对所得谱带的聚类分析表明，聚类结果与地理来源没有明显的联系，这可能正反映了近时间内地区之间枳资源的频繁交流。     

Genetic diversity in ecotypes of Tunisian date-palm (Phoenix dactylifera L.) assessed by AFLP markers

Summary

Amplified fragment length polymorphisms (AFLP) were used successfully to survey genetic diversity in 40 ecotypes of date-palm (Phoenix dactylifera L.) collected from oases in Tunisia. Six primer pairs were screened to assess their ability to detect polymorphism in this tree crop. As a result, a total of 428 AFLPs have been generated and used to estimate genetic distances which ranged from 0.07 – 0.63. A large, and typically continuous, range of genetic diversity characterises Tunisian date-palm germplasm. In addition, the UPGMA dendrogram derived from these data exhibited two clusterings of ecotypes independent of their geographic origin or the sex of the trees. These data corroborate the hypothesis of the origin of date-palm domestication being in Mesopotamia. Moreover, taking into account the high percentage of polymorphic bands, together with their resolving power (Rp), all the primer pairs tested contributed to the discrimination of date-palm genotypes, suggesting that the AFLP method is efficient in assessing genetic diversity in this crop. The data are discussed in relation to the use of AFLP molecular markers in the management and improvement of date-palm.     

仁用杏品种SRAP遗传多样性分析及指纹检索系统的开发

利用SRAP标记对24份仁用杏品种进行了遗传多样性分析。15对引物共扩增出280条带,其中241条为多态性谱带,多态性比率为85.34%;每个引物组合扩增谱带数在13 ~ 24条之间,平均为18.7条;多态性信息含量（PIC）在0.28 ~ 0.65之间,平均为0.51。基于引物Me4-Em4扩增的多态性谱带,构建的指纹检索系统可以区分24个仁用杏品种。根据SRAP扩增结果,利用UPGMA法构建树状聚类图,在相似系数为0.70处可将24个仁用杏品种分为4组,聚类结果与形态分类基本一致。