S_allele identification and genetic diversity analysis of apricot cultivars

In this study, the sexual incompatibility and S-allele diversity of 24 Turkish apricot cultivars, Paviot and Sak?t-1 as parents and 127 F₁ progenies were identified by polymerase chain reaction (PCR) and sequencing techniques. Additionally, genetic diversity and relatedness among the 24 cultivars were determined using 18 simple sequence repeat (SSR) markers from the genus Prunus. PCR for S_alleles identified nine different S-RNase alleles in the 24 apricot cultivars, namely S_c, S₁, S₂, S₈, S₉, S₂₀, S₂₄, S₅₂, and S₅₃. All primers amplified only one S_allele in the cultivars Adilcevaz-1, Adilcevaz-3, Ethembey, Pasamismisi, Canakkale, and Soganci. Most of the Turkish cultivars were self-incompatible. The S_c allele was present in only three cultivars (Canakkale, Ethembey, Imrahor) that are, therefore, self-compatible. The S_alleles of cultivars Paviot and Sak?t-1 displayed homology with the S_c, S₂ and S₂₀, and S₅₂ alleles. In the 127 F₁ genotypes, the two S_alleles of Paviot were inherited by roughly half of the offspring, while about 76% of the offspring inherited the S₅₂ allele from Sak?t-1, and less than 24% inherited S₂₀. The amplification using all SSR 18 primers was successful and produced 128 polymorphic alleles with an average of 7.11 alleles per locus. Among the apricot cultivars studied, expected heterozygosity (He) ranged from 0.33 to 0.72, observed heterozygosity (Ho) ranged from 0.42 to 1.00, PIC values were between 0.28 and 0.89, and similarity rates were between 0.30 and 0.68. The cultivars Levent and Ozal were genetically closest (0.68) while cultivars Sak?t-3 and Soganc? were the most distinct (0.30).     

167份西瓜种质材料的遗传多样性分析

以来自世界各地的167份西瓜种质资源为材料,利用形态学标记和SSR标记,结合多样性分析、聚类分析和主坐标分析等方法,对其遗传多样性进行了系统全面的研究。结果表明,29个表型性状Shannon多样性指数的变化范围是0.54~2.03,平均值为1.50,其中质量性状平均值为1.16,数量性状为1.71;18个数量性状的变异系数变化范围为16.43%~90.73%,平均值为34.12%。不同地区西瓜种质遗传多样性比较结果显示,非洲地区的种质遗传多样性丰富,亚洲、北美洲次之,南美洲和欧洲的遗传多样性相对较低。同时,利用22对SSR标记对西瓜种质进行了遗传多样性分析,平均每对引物扩增4.86个等位基因,平均有效等位基因数(Ne)是2.20,平均Shannon多样性指数(I)为0.94,平均观测杂合度(Ho)为0.27,平均期望杂合度(He)为0.52,平均多态性信息含量(PIC)为0.45。进一步分别利用表型标记和SSR标记对167份西瓜种质材料进行分类,结果显示SSR标记的稳定性强,能够很好的对种质材料进行分类。     

樱桃SRAP-PCR体系优化及其遗传多样性分析

选取亲缘关系较远的3个不同基因型樱桃资源为试材,对影响SRAP标记PCR反应的模板、Mg2+、dNTPs、Taq酶及引物浓度进行了优化,建立了适合于樱桃SRAP标记的扩增体系。反应体系具体为:模板DNA75ng,dNTPs0.2mmol·L-1,Mg2+2.5mmol·L-1,引物0.3μmol·L-1,Taq酶1.0U,反应总体积20μL。采用优化的扩增体系,对45个樱桃种质材料进行了遗传多样性分析,筛选8对扩增清晰且多态性高的引物组合,检测位点共227个,其中多态性位点192个,占84.6%。应用NTSYS-pc软件进行聚类分析(UPGMA),结果表明45个樱桃品种可分为欧洲甜樱桃和中国樱桃2大类,品种间遗传相似系数在0.52～0.98;其中中国樱桃与甜樱桃种间的相似系数最小,表明2类种质具有不同的遗传背景;而组群内的不同品种资源表现了较高的遗传相似性。SRAP分子标记的聚类分析揭示了樱桃品种间亲缘关系与地理分布以及来源相关。     

广东果梅种质资源遗传多样性的AFLP分析

为了从DNA分子水平上探讨广东境内果梅种质资源遗传多样性状况,利用荧光标记AFLP技术,对采自广东境内的57份果梅种质进行研究,筛选出适于果梅AFLP分析的8对Mse Ⅰ和EcoR Ⅰ引物组合,这些引物可以将供试材料完全分开.利用8对引物组合对57份果梅种质进行总基因组DNA水平上的多态性检测,得到了清晰的多态性指纹图...     

不同株系菠萝蜜染色体倍性及基因组大小分析

【目的】为了解菠萝蜜基因组特点,研究测定了10份菠萝蜜株系的倍性和基因组大小。【方法】分别以番茄和‘妃子笑’荔枝细胞核DNA含量为内标,Otto为提取液,采用流式细胞术(FCM)测定经碘化丙啶(PI)染色的内标和菠萝蜜细胞核混合样品发出的PI荧光强度,通过比较内标和菠萝蜜DNA峰值的荧光均值比值关系,计算出不同株系菠萝蜜的倍性和基因组大小。【结果】所测的10份菠萝蜜株系都为二倍体,干苞菠萝蜜株系的基因组在(1 243.75±9.21)~(1 540.63±3.55)Mb,湿苞菠萝蜜株系的基因组约为(1 902.87±8.08)Mb。【结论】不同菠萝蜜株系之间基因组大小存在显著差异,且湿苞株系大于干苞株系。     

Commercial Iranian olive cultivars: morphological traits,molecular diversity,and genetic structure

Iran is considered to have a unique gene pool of different fruit and nut species including olive (Olea europaea L.). In this study, we used 22 previously developed microsatellite (simple sequence repeat; SSR) markers for olive to evaluate the level of genetic variation and to produce identification keys for 63 Iranian accessions of olive belonging to 17 groups of cultivars. Based on morphological features, the number of flowers per inflorescence, fruit weights, endocarp weights, oil percentages, and flesh weights per endocarp had the highest coefficient of variation values, indicating the large extent of morphological variability among the 63 Iranian olive cultivars studied. All 22 microsatellite (SSR) markers revealed a high level of polymorphism, with a mean polymorphic information content (PIC) value of 0.511. Analyses of genetic structure among the 63 olive accessions were carried out using model-based methods, which showed a tendency for geographical clustering. Ten SSRs out of the 22 were successful for the identification of unique ID keys for 52 of the 63 accessions. In most cases, there was disagreement between the molecular data and the morphological data. These results could be used to reconstruct and maintain a collection of olive for future breeding programmes.     

板栗野生居群遗传多样性研究

使用从板栗中提取的10个微卫星引物对4个中国栗野生居群的69个个体进行扩增,共检测到84个等位基因,每个位点的等位基因数为4～13,每位点的平均等位基因数目为8.4个,平均有效等位基因数(Ne)为4.998,平均期望杂合度(He)为0.777,平均PIC值为0.739,Nei’s多样性指数(h)为0.771,以陕西汉中居群具有最高的遗传多样性。遗传分化系数Gst仅为0.141。UPGMA树状聚类图表明4个野生群体被分为两部分,其中云南、四川、安徽3个居群距离相近位于同一组,此结果与PCA分析结果一致,2种方法均显示居群间的遗传关系与实际地理分布不完全相关。     

一组柑橘嫁接嵌合体及其嫁接亲本的形态学、遗传学研究

对一组柑橘周缘嵌合体(Ekuliku)及其嫁接亲本(Nankan和Hamlin)的叶片和果实进行了形态学、生理学及遗传学分析。一般柑橘嵌合体的不同部位由不同亲本特定的组织层发育而来。实验结果表明,嵌合体果实的特性并不与其同层亲本的特性完全一致,嵌合体叶片横截面的结构也与亲本存在较大差异,说明在细胞层间存在着一定的相互作用。用127种随机引物和1对STS特异性引物对嵌合体和嫁接亲本作遗传学分析,其中8种引物能得到特异性条带,即只在嵌合体和其中的1个亲本中检测到;并且这8个引物都有很高的重现性。随即,对这些特异性条带作了克隆、测序和同源性比较分析。     

Comparative analysis of genetic diversity in Tunisian apricot germplasm using AFLP and SSR markers

Eighty-one accessions representing apricot germplasm in Tunisia were collected from different areas of cultivation and fingerprinted using amplified fragment length polymorphism (AFLP) and microsatellites (SSR) markers. A total of 339 polymorphic markers were revealed using 5 AFLP primers combinations and 24 SSR loci. AFLP and SSR markers expressed a high level of polymorphism allowing the distinction of the accessions with an efficiency coefficient of discrimination of 100% for AFLP and 97% for SSR markers. Genetic diversity structure was assessed with AFLPs and SSRs markers separately then with combined matrix data by the help of hierarchical clustering elaborated using Wards method based on Nei and Li (1979) distances. Comparison of the obtained dendrograms revealed a phylogeographic structure into two major groups with significant conservation between the observed subgroups in relation with the geographic origin of the accessions. The relative efficiency of the markers in determining the genetic relationships among apricot accessions has been assessed and a combination of AFLPs and SSRs markers was the most effective. In addition, Mantel test based on genetic distances indicated highly significant correlation between AFLP-SSR data and each of the AFLP and SSR ones, with Pearson correlation values of r = 0.873 and r = 0.692, respectively, revealing the higher efficiency of the combination of both molecular techniques (AFLP and SSR) to estimate the levels of genetic variability among apricot germplasm.     

杨桃遗传多样性的形态特征与RAPD标记的相关性分析

【目的】为指导杨桃种质资源的引进以及良种选育提供科学依据,【方法】采用形态标记数量聚类分析和RAPD分子标记相结合,对广东地区10份杨桃品种资源进行遗传多样性分析,并将形态学聚类和RAPD分子标记聚类结果加以对比。【结果】在所观察的12个形态性状中,变异系数为14.08%~49.71%,平均变异系数为25.38%。从100条RAPD随机引物中筛选出10个引物,共扩增出58条带,其中53条为多态性带,多态率达93.02%。聚类分析结果表明,形态标记和RAPD标记都可依果实风味将供试材料分组,即甜味和酸味。2种标记方法的相关系数为r0.01=0.685 3,达到显著水平。【结论】杨桃具有丰富的遗传多样性,2种方法聚类结果相似,且相关性高,具有较高的可靠性。     

甘肃省梨地方品种资源遗传多样性及亲缘关系SSR分析

为了解甘肃省不同生态区地方梨(Pyrus L.)品种资源的遗传多样性及亲缘关系,采用28对SSR引物对119份甘肃省梨地方品种或类型进行鉴定分析。供试28对SSR引物中有7对引物具有较好的多态性,7对引物共扩增出49条带,其中多态性条带28条,平均多态性位点百分率为63.27%;观察杂合度(H0)介于0.068~0.809,平均为0.591;平均多态性信息含量(PIC)(变异范围)介于0.365~0.680,平均为0.545;香农指数(I)介于0.713~1.482,平均为1.094;引物NH005b、NH007b及NH008b多态性较好,品种鉴定效率较高。所有材料遗传相似系数分布在0.074~1.000,平均为0.597;聚类分析显示,7对SSR引物可将119份供试梨品种资源在遗传相似性系数1.076处分为7组,主成分分析和UPGMA结果基本一致;群组内各品种资源由于其遗传相似性,更多地被聚在一起,但主成分分析并没有很明显地被分为多个类群或者分成几个主成分。研究结果揭示了甘肃省梨地方种质资源具有丰富的遗传多样性和广泛的遗传变异,还对根据形态特征无法确定归属的梨品种据其相互间的亲缘关系进行了适当归类,为进一步分类和利用甘肃省梨地方资源提供了理论依据。     

新疆野苹果资源遗传多样性SSR分析

采用SSR标记技术对新疆野苹果7个天然居群180份样品进行了遗传多样性分析,旨在为新疆苹果的杂交育种及优良品种/品系的选育提供理论依据。13对SSR引物共扩增出119条多态性条带,各居群内的多态性位点比率为79.84%~92.25%;居群水平上新疆野苹果的遗传多样性变化趋势基本一致,其中新源居群的有效等位基因数(Ne)、Nei's基因多样性(H)和Shannon信息指数(I)在7个居群中最大;种级水平上的Shannon信息指数(I)为0.551 6,居群内为0.468 9;新疆野苹果居群的变异主要存在于居群内部,居群间和居群内遗传多样性分别为15%和85%,说明新疆野苹果居群的变异主要存在于居群内部。     

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.     

Using bulked AFLP analysis to assess genetic diversity in Echinacea species

Echinacea is an allogamous genus, thus its cultivars or populations are genetically heterogeneous. Using amplified fragment length polymorphism (AFLP) to estimate the genetic diversity of Echinacea is generally limited by the large number of individual plants and the higher cost that need to be processed. In the present study, effectiveness of several sizes of DNA bulking (10, 15, 20, 25 and 30 individuals) with 20, 36 and 55 primer pairs was compared using AFLP in determining the genetic diversity of Echinacea species. The results indicated that the use of bulked DNA-based AFLP analysis by using the selected eight primer pairs was capable of detecting genetic diversity between the tested Echinacea species, provided that the potential presence of low frequency variants was ignored and a possible bias in the estimates of genetic similarity was accepted. The assessments showed that a bulk of 15 individuals could detect sufficient AFLP variations at most genomic sites. Additionally, 20 primer pairs could generate sufficient polymorphic fragments to achieve high resolving power of AFLP for the tested Echinacea species.     

四川干旱干热河谷核桃资源遗传多样性分析

利用AFLP-银染技术,对四川干旱半干旱河谷流域的4个核桃居群和干热河谷流域的2个铁核桃居群共46个样品进行遗传多样性与亲缘关系分析。选择EcoRⅠ/MseⅠ酶切组合,选用多态性高、分辨力强的引物组合E32/M48、E32/M61、E33/M61、E33/M62、E35/M74、E41/M48分别对供试材料的基因组DNA进行选择性扩增,共扩增出475个遗传位点,其中332个多态位点,多态率为69.89%;核桃群体所检出的位点平均有效等位基因数Ae、Nei’s基因多样度H、平均Shannon信息指数I分别为1.2972、0.1745、0.2629,而铁核桃群体分别为1.2758、0.1616、0.2422;从种级水平来看,2物种内部居群间的遗传分化系数Gst分别为0.3794,0.2252,表明核桃群体与铁核桃群体的变异主要存在于居群内,分子方差分析(AMOVA)的结果与以上分析结果一致;主成分分析(PCA)结果与聚类分析的结果相符;从Nei’s遗传距离和遗传一致度分析了各供试资源间的亲缘关系;经SPSS分析表明居群间的遗传距离与地理距离之间存在极显著相关性(R2=0.847**)。从研究结果可得出以下结论,物种的繁育系统、居群间的基因流、地理隔离是该区域核桃及铁核桃居群遗传结构形成的主要原因。     

Development of new microsatellite markers for molecular diversity analysis of Citrus species

Summary

Thirty microsatellite loci for further genetic analysis of Citrus species were developed by constructing a microsatellite-enriched library using capture with streptavidin-coated magnetic beads and used to assess genetic diversity in 40 Citrus accessions. In total, 150 alleles were detected, with an average of five alleles per locus. The average gene diversity and polymorphism information content values were 0.58 and 0.52, and ranged from 0.35 – 0.74 and from 0.32 – 0.70, respectively. Values for the observed and expected heterozygosities ranged from 0.13 – 1.00 and from 0.36 – 0.75, respectively. Fifteen loci deviated from the Hardy-Weinberg equilibrium (P < 0.05). The mean similarity coefficient among accessions was 0.5906. Based on the UPGMA algorithm, two main groups were successfully identified. These new microsatellite markers can be used to further investigate the genetics of, and phylogenic relationships in Citrus spp.     

菠萝蜜果实中糖分积累特征及相关代谢酶活性分析

【目的】探究菠萝蜜果实中的糖分组成及含量积累特征。【方法】以菠萝蜜品种‘马来西亚1号’为材料,利用高效液相色谱法分析不同发育时期果肉中蔗糖、果糖和葡萄糖含量,应用分光光度计法检测淀粉含量及相关代谢酶活性。【结果】菠萝蜜成熟果实中蔗糖含量最高,超过总糖含量的70%,主要在果实采后成熟期积累。果糖和葡萄糖含量接近,各占总糖的15%左右。淀粉是菠萝蜜果实中重要的非结构性碳水化合物,在果实成熟采摘前积累、后熟期间降解。蔗糖酸性转化酶(AI)为主要的蔗糖水解酶,与蔗糖含量呈显著负相关(相关系数为-0.602*)。蔗糖合成酶(SUSY)、蔗糖磷酸合成酶(SPS)在后熟阶段活性最高,与蔗糖含量呈显著正相关(相关系数分别为0.915**、0.997**),参与蔗糖的合成。腺苷二磷酸葡萄糖焦磷酸化酶(AGPase)在果实发育中期活性最高,与淀粉含量呈显著正相关(相关系数为0.900*)。α-淀粉酶在后熟阶段活力最高。【结论】蔗糖是‘马来西亚1号’菠萝蜜果肉的主要糖分,蔗糖的积累与AI活性呈显著负相关,与SUSY、SPS活性呈显著正相关,主要在后熟期积累。淀粉是菠萝蜜果实发育的主要非结构性碳水化合物。淀粉的积累与AGPase活性呈显著正相关,在果实成熟前合成、后熟期间降解。     

TP-M13-SSR技术在梨遗传多样性研究中的应用

利用在SSR扩增产物检测过程中的一种基于荧光测序技术的高通量低成本分析技术体系TP-M13-SSR (simple sequence repeat with tailed primer M13)对5个种25份梨种质资源材料进行了SSR标记的遗传多样性研究.利用NH013a和NH007b2对引物即可区分所有供试品种,5对...     

A PCR-based assessment of genetic diversity,and parentage analysis among commercial mango cultivars and hybrids

Summary

Three different PCR methods [Random Amplified Polymorphic DNA (RAPD), Inter-Simple Sequence Repeats (ISSR), and Directed Amplification of Minisatellite DNA (DAMD)] were used to analyse genetic diversity and parentage among 20 mango cultivars, including 18 landraces and two hybrids (‘Amrapali’ and ‘Mallika’). These hybrids together with a third hybrid (‘Ratna’), and an out-group species (Mangifera sylvatica) were also analysed for parentage. Fifteen, seven and four primers were used to amplify a total of 158, 69 and 59 distinct DNA fragments by RAPD, ISSR and DAMD, respectively. Of these, approx. 85%, 64% and 90% were polymorphic, respectively. Genetic distances between pairs of mango cultivars were measured separately by each method and depicted graphically as a Neighbor Joining (NJ) tree. The three methods revealed different groupings of cultivars and hybrids. A NJ tree based on the cumulative data from all methods correlated well with the parentage of the mango hybrids, and the grouping of cultivars on a regional basis. Genetic markers likely to be associated with important agronomic traits were identified by further analysing the hybrids, with their respective parents, using all three methods. On the basis of the highest number of polymorphic bands observed (90%), DAMD was judged to be the best method with which to analyse mango germplasm.