基于SSR标记的核桃种质资源遗传多样性与遗传结构分析

以58份核桃品种(优株)为试材,利用荧光SSR分子标记方法评价了58份种质资源的群体遗传多样性,分析其遗传结构特征,构建NJ聚类图,以期准确了解58份核桃种质的遗传背景和亲缘关系,为核桃属种质资源的合理利用、遗传资源的保存以及开发合理的育种方案和策略提供参考依据。结果表明：1)试验筛选出的15对SSR引物具有较高的多态性,共扩增出109个多态性等位基因(Na),平均每对引物的观测等位基因数(Na)为7.267,平均有效等位基因(Ne)3.069,平均观测杂合度(Ho) 0.519,平均期望杂合度(He)0.653,平均多态性信息量(PIC)0.603 5,平均香农指数(I)1.327。2)58份核桃品种(优株)间遗传距离范围为0.181～0.742,变幅为0.561,品种间遗传距离≥0.5的比例为44.19%,≥0.6的比例为13.5%,遗传基础较宽广,遗传信息丰富,遗传多样性水平较高。3)基于Nei遗传距离聚类分析,58份核桃品种(优株)可聚成3个类群,聚类结果与表型性状、亲缘来源有关,与地理位置相关度不大。4)群体遗传结构分析中,Q>0.6占比81.03%,基因混合来源比例较...     

核桃种质资源遗传多样性的RAMP标记分析

以核桃嫩叶为试材,对34份核桃种质资源进行了RAMP技术分析研究,以期建立基因组DNA的有效提取方法。结果表明:利用11对引物共扩增出185个条带,其中多态性条带142个,多态性条带比率62.5%~93.3%;34份核桃样品间的相异系数0.0274~0.4848。表明34份核桃种质资源具有较丰富的遗传多样性,用RAMP进行核桃资源的DNA指纹分析可行;核桃的遗传基础具有一定的地域特征,"石门核桃"种群在遗传关系上是相对独立的栽培群体。     

耧斗菜野生资源的AFLP遗传多样性分析

以包括22份野生资源的5个耧斗菜物种为试材,采用AFLP分子标记方法,分析了中国耧斗菜野生资源的遗传变异和遗传多样性,为系统鉴定耧斗菜野生资源奠定基础。结果表明:10对引物共扩增出293个多态性位点,多态性位点比率为62.5%。Shannon多样性和He值分析结果表明,5个物种的遗传多样性大小顺序为无距耧斗菜耧斗菜尖萼耧斗菜直距耧斗菜华北耧斗菜。遗传距离分析结果表明,耧斗菜与尖萼耧斗菜遗传距离最小,尖萼耧斗菜与直距耧斗菜遗传距离最大。     

茶树品种资源遗传多样性的AFLP研究

 利用AFLP - 银染分子标记技术, 对40个茶树品种(系) 进行遗传多样性与亲缘关系分析。选用多态性高、分辨力强的引物组合E-ACG/M-AAC、E-AGG/M-AAG与E-AGG/M-AGT分别对供试材料的基因组DNA进行扩增, 共获得226条清晰可辨的带, 其中多态性带207条, 多态位点百分率为91.59% ,这表明供试品种资源在DNA水平上酶切位点的分布存在广泛的变异。40份资源所检出的位点平均有效等位基因数、平均基因多样度、平均Shannon信息指数, 分别为1.59 ±0.09、0.35 ±0.04、0.53 ±0.05。应用SPSS软件计算遗传距离介于0.13～0148之间, 平均为0.32; UPGMA法将40份资源分成4个类群, 从相似性系数分析了各品种资源间的亲缘关系。     

菊花品种资源遗传多样性的AFLP分析

以AFLP—银染分子标记技术, 对45个菊花品种进行了遗传多样性和亲缘关系分析。选用10个多态性高、分辨力强的E + 3 /M + 3引物组合分别对供试材料的基因组DNA进行扩增, 共获得486条清晰可辨的条带, 其中多态性带451条, 平均每个引物组合可检测出4511个多态性位点, 多态性位点百分率高达92.80% , 这表明供试品种资源在DNA水平上酶切位点的分布存在广泛的变异。应用DPS软件计算供试品种遗传距离界于0.36000～0.86237之间, 平均为0.611185; UPGMA分析将45份资源分成6个类群,从相异性系数分析了各品种资源间的亲缘关系。     

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...     

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。     

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。     

珙桐遗传多样性的AFLP 分析

 采用7对AFLP引物对中国珍稀濒危植物珙桐（Davidia involucrata Baill.）的6个天然居群和2个人工居群的229份材料进行扩增,共检测到100 ~ 500 bp的位点506个,其中多态性位点488个,多态性位点百分率96.44%;各居群多态位点百分率为52.77% ~ 79.25%,平均为67.00%。在物种水平上,珙桐的Nei’s基因多样性H_s = 0.3429,Shannon’s遗传多样性信息指数I = 0.5107。而居群水平上,各居群的Nei’s基因多样性介于0.2229 ~ 0.3389,各居群的平均Nei’s基因多样性H_s = 0.2748,各居群Shannon’s遗传多样性信息指数（I）介于0.3219 ~ 0.4877,平均Shannon’s遗传多样性信息指数I = 0.3995。居群间基因间分化度（G_st）为21.60%,基因流N_m为1.8。说明珙桐的遗传多样性较高,这8个居群间存在一定的遗传分化和基因流动,但遗传分化主要存在于居群内。在珙桐与其变种光叶珙桐之间未找到差异标记。     

用AFLP标记分析广西龙眼种质遗传多样性

选用10对AFLP引物组合分析了广西38个龙眼品种（优良单株）和两个龙眼近缘亚种龙荔单株,以及10个来自国内外其他产区龙眼品种的遗传多样性。在所扩增的570条AFLP主带中,有485条具有多态性,占85.1%。所用引物可将供试的品种（优良单株）区分开来,供试材料的遗传相似系数在0.39（两个亚种间）到0.98之间。根据遗传相似系数（UPGMA, N-J）得到的分类树状图与传统方法的分类结果相类似。结果表明,广西龙眼种质具有较广泛的遗传多样性。     

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.     

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.     

桃野生种和地方品种种质资源亲缘关系的AFLP分析

利用AFLP技术对94份野生桃种质资源进行了DNA多态性分析,从64对E+2/M+3引物组合中筛选出9对引物用于扩增基因组DNA,共获得清晰可辨的236个标记,其中多态性标记为141个,多态性检出率为59.9%。UPGMA聚类结果显示,在遗传距离约为0.3267时,普通桃、陕甘山桃、光核桃3个种被聚为一类,与这3个种相近的为新疆桃,其次为甘肃桃,最远为山桃(白花山桃、红花山桃)。在遗传距离0.2178处,五月鲜扁干、白黏胡桃和珲春桃3份材料各自聚为一类。在遗传距离约0.1997处,普通桃又可明显分为4个组:组I包括原产于南方和西北地区的地方品种,且大部分南方蟠桃主要聚在此类;组II仅有油桃(甘肃酒泉)一个野生材料;组III包括野生毛桃和大部分原产于北方的地方品种,北方硬肉桃主要聚在这一类;组IV由白碧桃和白花山碧桃2个观赏桃品种组成。     

Genetic diversity and population genetic structure of pomegranate (Punica granatum L.) in Iran using AFLP markers

Pomegranate (Punica granatum L.) is one of the oldest known edible fruits. It is native to Iran and spread from Iran to other areas. In this study amplified fragment length polymorphism (AFLP) was used to detect intra- and inter-population genetic diversity of pomegranate. A group of 67 accessions belonged to 4 populations from Iran was studied using eight primer combinations. A total of 221 scorable bands were amplified, of which, 118 (54.13%) were polymorphic. Resolving power (Rp) ranged from 5.70 to 9.21, and the average of polymorphism information content (PIC) per primer pair was 0.40. According to Nei's gene diversity and allelic statistics, Isfahan population had a highest genetic diversity (H = 0.3646, I = 0.5327, N_e = 1.6467). Coefficient of gene differentiation between populations (GST) was 0.124, indicated that mainly proportion of genetic variation (87.6%), was within populations and the remaining (12.4%) of the variation was among populations that, also supported by analysis of molecular variance (AMOVA). The gene flow (Nm) varied from 0.969 to 10.404 between pair-wise populations and was 3.504 among all of the populations. The Jaccard similarity coefficient between individuals ranged from 0.26 to 0.88. The UPGMA dendrogram clustered all 67 accessions into 6 groups. In some cases accessions from same region were grouped together but in most cases, there was gene exchange. To study the genetic relationships among populations, a principal coordinate analysis (PCoA) based on Nei's genetic distances was performed. Results of this study showed that AFLP marker can be a useful tool for investigating the genetic diversity of pomegranate genotypes.     

菠萝种质鉴定及亲缘关系的AFLP分析

从FISH-AFLP试剂盒(PSTI/MseI型)64对引物中,筛选出8对AFLP选择性引物P-GAA/M-CTA、P-GAC/M-CTG、P-GAC/M-CTT、P-GAG/M-CAT、P-GAG/M-CTA、P-GAT/M-CAT、P-GAT/M-CTA、P-GAT/M-CTC,对39份菠萝种质进行了AFLP分析,都得到了清晰的多态性指纹图谱。8对引物共获得454个遗传位点,其中多态性位点332个,多态性比例平均为73.1%。UPGMA聚类表明,各种质间的相似系数为0.73～0.98,说明39份菠萝种质遗传关系相对较近。依相似系数0.80的水平,将供试的39份菠萝种质分为4类。研究结果可以作为菠萝分类的理论依据和基础。     

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。     

安徽砀山酥梨自然保护区梨种质资源AFLP分析

为探讨安徽砀山酥梨自然保护区梨种质资源遗传多样性和鉴定砀山酥梨营养系变异,以该保护区内27个梨资源代表类型为试材,利用AFLP分子标记技术开展研究。运用从36对选择性扩增引物中筛选的3对引物组合进行扩增,每对引物可扩增出50条以上的清晰条带,扩增片段长度为30～600bp,9个引物组合共扩增出清晰可靠的DNA谱带540条。其中498条为多态性条带,平均多态性比率为92.2%。聚类分析表明,当遗传距离阈值为39.2～40.1时,全部供试生物型可分为6类:第1类为砀山酥梨及其变异类型和原产砀山的地方品种、杂交种;第2类包括雪花梨、鸭梨、紫酥、慈梨、早美酥、丰水、新高,白梨和砂梨聚在了一类;第3类包括库尔勒香梨、红香酥;南果梨、红巴梨、杜梨各为一类。与砀山酥梨相比,砀山酥梨的8个营养系变异类型均扩增出了特异带,特异条带出现的数量为5～15条不等。说明安徽砀山酥梨自然保护区梨种质资源遗传多样性丰富;砀山酥梨8个营养系变异类型均在DNA水平发生了不同程度的突变。     

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.     

梨抗黑星病基因的AFLP分子标记

为了解梨黑星病抗性遗传规律并为梨黑星病抗性育种早期选择提供理论依据,以新苹梨(梨黑星病抗性品种)×雪花梨(梨黑星病易感品种)的杂交后代为试材,进行了田间自然感病和棚内接种梨黑星病(Venturia nashicolaTanak et Yamamota)病菌后的抗性调查,并进行了AFLP分子标记的研究。结果表明,棚内接种梨黑星病菌后,杂交后代抗感分离规律符合双假测交的遗传规律;研究建立的AFLP分子标记体系可以用于梨黑星病的抗性基因标记,并初步找到了与梨黑星病抗性基因相关的分子标记。