中国南瓜海南农家品种资源遗传多样性和亲缘关系研究

为系统评价中国南瓜海南农家品种资源,通过表型变异结合ISSR和SRAP分子标记技术,对中国南瓜海南农家品种进行遗传多样性和亲缘关系分析。表型分析结果表明,在所有观察的农艺性状中,除蔓性、生长势、果柄、果肉质地等4个性状外,其他性状在不同品种间均有不同程度的差异,平均变异系数为34.88%,其中果形的变异最大,部分农艺性状间存在显著的相关性;使用UPGMA法聚类分析可依据叶片大小、叶色和叶尖将28份材料划分为2个类群。ISSR标记和SRAP标记所揭示的遗传多样性水平基本一致,均表明供试品种间存在较大的遗传变异。基于ISSR标记,所有供试品种间的Nei's基因多样性指数H为0.1943,多态性条带百分率PPB为62.90%;基于SRAP标记,H=0.2258,PPB=61.50%;2种标记的UPGMA聚类结果存在显著的相关性,均可将供试品种分为5类,但是其聚类结果与表型分析结果之间相关性较低。综合2种标记的扩增结果,各品种间的平均遗传相似性系数GS值为0.7741,海13和海17亲缘关系最近,而海30与海18遗传基础差异最大。供试中国南瓜海南农家品种间存在较丰富的遗传变异,ISSR和SRAP标记均能有效应用于中国南瓜海南农家品种遗传多样性和亲缘关系研究。研究结果为中国南瓜海南农家品种保护和创新利用提供了科学依据。     

广西地方葛根种质资源遗传多样性的SCoT分析

为了解广西地方葛根种质资源之间的亲缘关系,本研究以SCoT分子标记对44份广西葛根种质资源的基因组DNA进行PCR扩增,并进行遗传多样性分析。结果表明,25条SCoT引物共扩增出223个条带,其中具有多态性的条带有194条,平均每条引物获得7.76个多态性条带,多态性比例为86.99%。聚类分析表明,44份葛根种质资源的遗传相似系数在0.587~0.982之间。在系数0.65处,44份葛根分为两大类,37号材料单独成为一类;在系数为0.74处,可聚为六类。综上所述,SCoT分子标记适用于葛根种质资源遗传多样性分析,本研究结果为广西葛根种质资源鉴定评价和新品种选育奠定了一定的理论基础。     

南瓜栽培品种的SSR分子标记分析及农艺性状多样性研究

为了研究南瓜栽培品种的遗传多样性,本研究利用43个简单序列重复(SSR)分子标记,对35份南瓜育成品种及地方品种进行了分子标记分析,并调查了农艺性状。结果表明,43个SSR标记均能扩增出多态性条带,共检测到155个等位基因,平均每个标记能检测到3.6个等位基因,多态性信息含量(PIC)为0.130 8~0.775 4,平均值为0.487 2。利用非加权组平均法(UPGMA)进行聚类分析,结果表明35份材料可分为三大类,分别与中国南瓜、印度南瓜和美洲南瓜三个种吻合,且印度南瓜与美洲南瓜之间的亲缘关系较近。农艺性状调查结果表明,不同栽培种之间以及同一栽培种内的不同品种之间,都发现有农艺性状差别明显的情况。本研究为南瓜种质资源的保护、品种指纹图谱的建立及分子育种提供了理论依据。     

甘薯种质资源的SRAP鉴定及遗传多样性分析

为了拓宽甘薯育成品种的遗传背景,筛选优良亲本,提高育种效率,本研究利用SRAP标记对48份主要甘薯种质资源进行了遗传多样性分析.结果表明,随机选用的37对SRAP引物中29对引物具有多态性,引物多态性比率为78.4％,共获得126条多态性谱带,平均每对引物产生4.3条多态性谱带,表现出较高的多态性.48份种质材料的SRAP遗传距离为0.037～0.601,当遗传距离L1=0.46时,48份材料被聚为6个类群,包括1个复合大类群和5个独立类群,其中第Ⅰ复合大类群又包括7个亚类群,聚类结果与系谱吻合性较好.采用5个重要农艺性状对供试材料进行了聚类分析,L1 =3.20时,48份材料也可被聚为6个类群,聚类结果与依据SRAP标记聚类分析的结果差异较大.甘薯种质资源间的遗传差异与地理来源无必然联系.     

RAPD和ISSR标记对甜瓜种质遗传多样性的研究

利用RAPD和ISSR两种分子标记技术对37份甜瓜(CucumismeloL.)种质进行了遗传多样性研究.在供试材料中筛选到具有多态性的RAPD引物21个,ISSR引物10个(其中ISSR-2与ISSR-4等量混合组成ISSR-10引物对).RAPD引物共扩增出多态性带106条,多态性条带比率(PPB)为58.62%,平均多态信息量(PIC)为0.47;ISSR引物共扩增出多态性带73条,PPB值为65.51%,平均PIC值为0.53.根据两种标记的结果,采用UPGMA聚类分析,将供试材料分为两大类群野生甜瓜和栽培甜瓜;栽培甜瓜又分为厚皮甜瓜和薄皮甜瓜两大亚群,各野生甜瓜种质之间的遗传距离较大,这与其分类地位基本一致.两种分子标记的分析结果呈极显著正相关(r=0.62>r=0.01).研究表明,RAPD和ISSR标记可用于甜瓜种质遗传多样性的研究.     

广西地方食用木薯种质资源遗传多样性分析

为研究广西地方食用木薯材料的遗传多样性,以48份广西地方食用木薯为试验材料,分析其变异系数、遗传多样指数、聚类分析、多态性和遗传相似性系数。结果表明,收集的木薯材料的平均表型变异系数为37.9%,平均遗传多样性指数为0.807,其中主茎分叉角度的变异系数最大,为86.7%,块根内皮颜色的遗传多样性指数最大,为1.842。13对SSR引物共扩增出118个条带,其中多态性条带为106个,多态性比率为86.23%;分子标记聚类结果发现,遗传相似性系数在0.415~1.000之间;通过对比发现表型聚类和分子聚类结果不一致,其中表型聚类发现类群划分与地理来源之间没有关联,但与株型性状有一定的关联;在遗传相似系数为0.62时,SSR分子标记聚类为两类材料,第一类材料地理分布无规律,第二类材料大多分布在桂南。本研究结果表明,广西地方食用木薯资源遗传多样性具有一定的丰富度,可为创制优异食用木薯种质资源和新品种选育奠定基础。     

利用RAPD和AFLP标记分析烟草种质资源的遗传多样性

从200条10 bp的RAPD引物中筛选获得28条多态性引物，对48份烟草材料的基因组DNA进行扩增。共获得184条DNA扩增片断带，其中多态性带86条，平均多态检出率为46.7%。用4对AFLP选择性扩增引物对48份烟草材料进行选择性扩增，共获得321条扩增带，其中174条具有多态性，平均多态检出率为54.2%。根据RAPD和AFLP标记的结果，采用UPGMA法进行聚类分析，两种方法获得了相似但不完全相同的结果，两者都可以将48份烟草资源分为两大类群，即黄花烟草(Nicotiana rustica)群和普通烟草(N.tobacum)群，普通烟草可进一步分为4组；48份材料的遗传距离变幅在1.4～11.0之间，其聚类结果与理论上所期望的结果基本一致，AFLP标记技术比RAPD标记技术能更好地从分子水平上揭示烟草种质资源的遗传背景、亲缘关系及演化规律。     

利用小麦SSR标记分析鸭茅种质资源的遗传多样性

使用小麦(Triticum aestivum)SSR引物和扩增程序,以中国春小麦(T. aestivum)品种为对照,利用SSR标记对来自国内外的45份鸭茅(Dactylis glomerata L.)种质资源进行遗传多样性研究.结果表明,20对引物扩增出295个条带,多态性条带为187条,多态性条带比率为61.15%,鸭茅种质资源的遗传相似系数范围为0.7848～0.9513.聚类分析和主成分分析将供试材料分为6大类,聚类结果不仅能反映鸭茅生态适应性特征与其生长发育状况及生产性能相关,还显示出国产鸭茅品种遗传基础较为狭窄.研究表明将小麦SSR引物用于检测鸭茅遗传多样性行之有效.     

16份石榴RAPD扩增产物的两种电泳方法检测及其序列特征

本实验以16个石榴品种为实验材料,筛选出10个重复性及多态性均较好的引物进行RAPD分析。分别采用琼脂糖凝胶以及聚丙烯酰胺凝胶(PAGE)电泳检测方法对PCR扩增结果进行检测并对其结果进行比较,结果显示,两种电泳方式均能得到较为清晰的扩增条带,且两种电泳方式获得的条带总数及多态性条带数均有所不同,琼脂糖凝胶电泳方法共检测出76条带,其中有43条为多态性谱带,多态性比率为56.4%;而在PAGE电泳方法共检测出123条谱带,多态性谱带数为87条,多态性比率为70.95%。PAGE电泳方法检测出的条带数约为琼脂糖凝胶电泳方法检测出条带数的1.5倍。基于两种电泳方法所得RAPD标记的多态性位点,利用NYSYS软件计算遗传相似系数,并构建遗传关系聚类图,分析结果显示,石榴遗传多样性丰富,两种电泳方法所得聚类结果大致相同,可以利用RAPD分子标记及两种电泳检测方法对不同数量的石榴进行分子水平的品种鉴定和遗传多样性的分析。同时通过对来自几个引物随机挑选的17个片段进行克隆,测序结果显示17个片段都是对应引物的RAPD扩增产物,其中有3条是编码蛋白的基因片段,表明了RAPD不仅扩增基因组上的非编码蛋白序列,同时也可以扩增编...     

用ISSR分析四川苎麻品种(系)间的遗传关系及雄性不育分子标记的建立

本文从100条ISSR引物中筛选出21条引物,对8个四川苎麻品种(系)间的遗传关系进行了分析。PCR扩增结果表明,21条引物在8份材料中共扩增出86条带,平均每条引物扩增出4.1条,其中多态性位点71个,各引物扩增出的位点数3~8个不等,平均每条引物可以检测到3.4个多态性位点。聚类分析和遗传距离分析结果表明,供试品种川7、川8与其亲本遗传距离较远。3个杂交品种(川7、川8、川9)均表现特有的偏父本遗传现象。此外,本研究用ISSR引物U835筛选到了1个雄性不育分子标记,并将其扩增产物克隆测序,结果表明该序列大小为658bp。根据该序列,此标记被转化成稳定的SCAR标记,可用于苎麻雄性不育分子标记辅助育种。     

Genetic variation in radish (Raphanus sativus L.) germplasm from Pakistan using morphological traits and RAPDs

Genetic diversity of 30 radish (Raphanus sativus L.) accessions was investigated at the phenotypic level with morphological characters and at the DNA level using the random amplified polymorphic DNA (RAPD) technique. Thirty-six morpho-physiological traits were recorded from seedling stage to harvest. The 31 primers used generated 202 RAPD bands, of which 158 (78.2%) were polymorphic. Multivariate procedures were used to classify the germplasm on the basis of phenotypic traits and RAPD fragments. Dendrograms were generated for the Euclidean distance from the morphological data and the Nei's genetic distance from the RAPD markers. Phenotypically, all the accessions were classified into four major groups corresponding to the different forms of cultivated radish. The morphological diversity existing within each of these groups suggested that they should be discriminated into the three botanical convarieties, sativusT (large-rooted), caudatus (pod-type) and oleifer (oilseed-type). Clustering of the accessions did not show any pattern of association between the morphological characters and the collection sites. Instead, landrace groups were associated with their morphological similarities and horticultural uses. On the other hand, the intra-specific genetic relationships of several accessions based on RAPD analysis were related primarily to their collection sites rather than to their phenotypic affinities. The level of polymorphism exhibited by the various convarieties could be exploited in genetic mapping populations to tag economically important traits. These genotypes also could serve as a useful germplasm source for root, leaf, pod and seed. This preliminary study of traditional radish landraces from Pakistan provides useful information regarding their horticultural potential.     

Genetic diversity of introduced and local Spanish persimmon cultivars revealed by RAPD markers

Persimmon culture has become an alternative for fruit growers in Italy, Israel and Spain. However, there is some confusion concerning cultivar identification in non-Asian countries were the species was introduced. Authenticating the identity of germplasm resources of persimmon would be a great value for breeding. Due to the floral biology of the persimmon species the expected genetic variability is high. RAPD markers were chosen for this study of persimmon germplasm resources in Spain. Nineteen markers were obtained from 8 primers. The average of 2.4 markers/primer was higher than that obtained in other woody fruit species. The availability of data referenced from isozymes and pomological traits revealed complete agreement among groups identified by RAPDs, isozymes and pomological traits. Results suggest that RAPD technology is adequate for fingerprinting persimmon and the data is consistent with persimmon's hypothetical origins, adaptation history and previous classification by isozymes or pomological traits. Many synonyms have been identified, mostly in cultivars from local surveys, suggesting that a few new genotypes arose in Spain since the species was introduced to Europe. By means of RAPD markers, a better identification of the European persimmon germplasm is available.     

大豆种质资源RAPD标记遗传多样性研究

为深入研究并充分利用野生大豆资源,本文利用RAPD分子标记对40份大豆材料加以分析,旨在从DNA分子水平上探索野生大豆、地方品种和育成品种之间的遗传多样性状况。结果表明:50个RAPD引物筛选出具有多态性且扩增条带清晰的引物38个,共检测出407条带,其中多态谱带309条,多态性程度为75.92%。每个引物可扩增出2～14条多态性带,平均产生多态性谱带8.1条;平均多样性指数为2.3377,变幅范围为0.5865～4.2133。遗传相似系数变幅范围为0.44～0.92,平均为0.75。野生大豆的多态比例(94.35%)、多样性指数(2.2336)分别高于育成品种(87.47%、1.7331)和地方品种(83.54%、1.6198)。遗传相似系数为野生大豆(0.6498)地方品种(0.7015)育成品种(0.7177),育成品种与地方品种间为0.6599,育成品种与野生大豆间为0.6487,地方品种与野生大豆间为0.6045。UPGMA聚类分析结果表明,40份大豆材料聚为6类,育成品种和地方品种各自聚为一类,野生大豆聚为4类。野生大豆特异等位基因数远远高于育成品种和地方品种二者的相加之和。本研究从分子水平上揭示了野生大豆与栽培大豆区别明显,宜作为一个独立的种,同时野生大豆变异幅度大,遗传基础广,是大豆育种实践中的优良基因资源。     

Marker-assisted sampling of the cultivated Andean potato Solanum phureja collection using RAPD markers

The potato crop originated in the Andean highlands where numerous farmer's varieties and non-cultivated wild species exist. An Andean potato collection is held in trust at the International Potato Center (CIP) to preserve the biodiversity of this crop and ensure the supply of germplasm for potato improvement worldwide. A core collection representing the biodiversity of the Andean potato germplasm is under construction using morphological, molecular, and geographic data. One of the eight cultivated potato species, Solanum phureja, has been genotyped using the RAPD technique. A protocol suitable for large germplasm collection genotyping has been developed to process numerous samples at reasonable costs. From 106 RAPD primers evaluated, we have selected 12 primers yielding 102 polymorphic markers, which unambiguously discriminated all 128 accessions but 2 that are possible duplicates. The S. phureja germplasm collected throughout the Andean countries appears to have a homogeneous genetic constitution. There was no clear geographic pattern as indicated by cluster analysis of the RAPD data. A sub-group of 20 accessions has been identified on the basis of the marker data and selected to maximize molecular (RAPD) variance and polymorphism. The probability of capturing equal amounts of marker polymorphism in this sub-group of 20 accessions by random sampling is less than 40%. This set accessions represents our first group of accessions that may constitute a core of the S. phureja collection. This tentative core will be challenged for diversity content by alternate markers and agronomic traits. Hence, the methodology for sampling less than 10% of the base collection, proposed for core collections by Brown (1989), can be based on molecular marker data provided cost-efficient fingerprints are developed.     

RAPD and ISSR fingerprinting in cultivated chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.) and its wild progenitor <Emphasis Type="Italic">Cicer reticulatum</Emphasis> Ladizinsky

Detection of genetic relationships between 19 chickpea cultivars and five accessions of its wild progenitor Cicer reticulatum Ladizinsky were investigated by using RAPD and ISSR markers. On an average, six bands per primer were observed in RAPD analysis and 11 bands per primer in ISSR analysis. In RAPD, the wild accessions shared 77.8% polymorphic bands with chickpea cultivars, whereas they shared 79.6% polymorphic bands in ISSR analysis. In RAPD analysis 51.7% and 50.5% polymorphic bands were observed among wild accessions and chickpea cultivars, respectively. Similarly, 65.63% and 56.25% polymorphic bands were found in ISSR analysis. The dendrogram developed by pooling the data of RAPD and ISSR analysis revealed that the wild accessions and the ICCV lines showed similar pattern with the dendrogram of RAPD analysis. The ISSR analysis clearly indicated that even with six polymorphic primers, reliable estimation of genetic diversity could be obtained, while nearly 30 primers are required for RAPD. Moreover, RAPD can cause genotyping errors due to competition in the amplification of all RAPD fragments. The markers generated by ISSR and RAPD assays can provide practical information for the management of genetic resources. For the selection of good parental material in breeding programs the genetic data produced through ISSR can be used to correlate with the relationship measures based on pedigree data and morphological traits to minimize the individual inaccuracies in chickpea.     

Genetic Relationships and Variation among Biotypes of Dallisgrass (Paspalum dilatatum Poir.) and Related Species Using Random Amplified Polymorphic DNA Markers

The genus Paspalum L. consists of more than 400 species. Around twenty-five informal groups of species are recognized in Paspalum and the Dilatata group is of special interest because its members are excellent potential forage grasses. Seventy-five germplasm accessions, representing 15 taxa, were analyzed using randomly amplified polymorphic DNA (RAPD). Polymorphisms were observed with twenty-two primers in the Dilatata group and 16 of those were analyzed. Four hundred and four different RAPD fragments were generated, resulting in an average of 25.2 bands per primer. Among the 404 markers analyzed, 48 (11.88%) were exclusive for the P. dilatatum Poir. biotypes, 31 (7.67%) were exclusive to taxa belonging to other groups included in this study, 28 markers (6.93%) were diagnosed for other species of the Dilatata group and 16 (3.96%), for natural hybrids. Extensive RAPD variation was found among the species studied. Inter- and intra-taxonomic polymorphisms were detected. A dendrogram based on the RAPD data shows some clusters corresponding to the same taxa. However, the biotypes of P. dilatatum do not form a cluster. The present work confirms that the RAPD technique can be used to determine genetic relationships between the taxa belonging to the Dilatata group.     

Determination of genetic variability by RAPD markers in cauliflower,cabbage and kale local cultivars from France

RAPD markers were used to analyse the genetic variability among a cole crop cultivar collection from France and to identify possible duplicate accessions for facilitating the germplasm multiplication. We surveyed 24 cauliflower, 24 cabbage and 48 kale populations using 62, 100 and 89 RAPD markers respectively on 40 seed bulk samples per accession. Genetic distances were calculated on the basis of these markers. Using the phylogenetic package PAUP we compared the genetic clusters obtained for the RAPD markers with the groups based on morphologic and agronomic data. For cauliflowers and cabbages a drastic selection by growers led to an extensive differentiation of morphologic and precocity traits related to geographic origin, which matches the grouping based on RAPD markers. Furthermore, RAPDs detected in some cases misclassification of accessions in the collection. Kales formed an homogeneous group probably due to extensive genetic exchanges leading to a continuous diversity. For the other crops, different horticultural types were separated in several subgroups by the RAPD markers suggesting different genetic origins. RAPD markers provide a rapid and informative approach to analyze the genetic variability in a collection. It can be applied to autogamous (spring cauliflower) or allogamous (winter cauliflower) crops, even in cases where the intra-population diversity is extensive (cabbages).     

Measurement of genetic dissimilarity in fieldpea (<Emphasis Type="Italic">Pisum sativum</Emphasis> L.) genotypes using RAPD markers

The present investigation was carried out on fifteen germplasm lines of Pisum sativum L. were used for characterization using Randomly Amplified Polymorphic DNA (RAPD) markers. While 12 random primers were taken, out of them 11 primers gave amplification. These primers gave a total of 133 bands out of which 106 were polymorphic. Genetic similarities of the RAPD profiles were estimated by using Jaccard’s coefficient with NTSYSpc 2.0 software. The similarity index values ranged from 0.263 to 0.793 indicating the presence of enormous genetic diversity at molecular level. A dendrogram generated by cluster analysis divided fifteen fieldpea genotypes into two Groups A and B. Major Group A have five genotypes and major Group B have nine genotypes.     

海南荔枝(Litchi chinensis Sonn.)主要栽培品种的RAPD分析

从80个随机引物中筛选出12条重复性好的多态性引物，对包括15个栽培品种在内的22份海南荔枝(Litchi chinensis Sonn.)资源进行RAPD分析，扩增的多态性片段比例为64.1%, 平均为31.1%; 遗传距离在0.0000~0.6180之间，平均0.3732。UPGMA聚类分析显示，22份资源主要分为2类，部分供试品种存在明显的同名异物现象。     

Relationships among different geographical groups,agro-morphology,fatty acid composition and RAPD marker diversity in Safflower (<Emphasis Type="Italic">Carthamus tinctorius</Emphasis>)

Patterns of geographical diversity, and the relationship between agro-morphological traits and fatty acid composition were assessed for 193 safflower (Carthamus tinctorius) accessions representing forty countries. Accessions were assigned to eight groups based on geographical proximity. Cluster and Principal Component analyses were performed to assess patterns of diversity among the accessions and to select the most distant accessions from each of eight groups for analysis of randomly amplified polymorphic DNA (RAPD) markers. There was a large amount of diversity for agro-morphological traits, fatty acid composition, and RAPD markers. Most correlations among different traits were rather low. Plant height showed a positive correlation with days to flowering (r = 0.63**). Palmitic acid was positively correlated with stearic acid and oleic acid values, and negatively correlated with linoleic acid (P < 0.01). Oleic acid and linoleic acid showed a strong negative correlation (r = −0.89**). The first three principal components together explained 59% of the variation, however, neither principal component analysis (PCA) nor marker analysis revealed a clear relationship between diversity pattern and geographical origin. Accessions from some geographical regions tended to group together, such as accessions from South Western Asia, Central Western Europe, and the Mediterranean region. The correlation between the morphological matrix and the genetic matrix based on RAPD markers was not significant (r = 0.027). Wide diversity in safflower germplasm indicates a considerable potential for improving this crop for both agronomic and quality traits.