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1.
RAPD技术及其在微生物学方面的应用   总被引:4,自引:0,他引:4  
198 0年 ,Botsein提出DNA限制性片段长度多态性 (RFLP)可以作为遗传标记 ,从此开创了直接应用DNA多态的新阶段。 80年代后 ,DNA多聚酶链式反应 (PCR)的发展 ,使直接扩增DNA的多态性成为可能 ,并在此基础上产生了许多种新型分子标记 ,诸如扩增片段多态性 (ALFR)、串联重复序列(VNTR)、单链构型多态性 (PCR SSCP)、序列特异扩增区域 (SCAR)、随机扩增多态性DNA(RAPD)等。而RAPD是较为突出的一种。RAPD是由Williams和Welsh在 1 990年各自独立发现的一种DNA多态检…  相似文献   

2.
西瓜抗枯萎病育种分子标辅助选择的研究   总被引:23,自引:1,他引:23  
将西瓜野生种质PI296341抗枯萎病生理小种1的抗性基因连锁的RAPD标记OPP01.700进行克隆、测序,Southern杂交证明此标记为1个单拷贝,并转化为SCAR标记,简化了SCAR扩增产物的检测技术。上述技术在抗病转育后代造反中得到了很好的应用,初步建立了西瓜抗枯萎病育种分子标记辅助选择技术系统。  相似文献   

3.
小麦抗白粉病基因Pm21的分子鉴定和标记辅助选择   总被引:26,自引:4,他引:26  
刘志勇  李洪杰 《遗传学报》1999,26(6):673-682
利用小麦抗白粉病基因Pm21的RAPD标记、SCAR标记和荧光源位杂交技术对小麦抗病育种材料中的抗白粉病Pm21基因进行了分子鉴定和标记辅助选择。  相似文献   

4.
DNA分子标记在柑桔中的应用   总被引:3,自引:1,他引:2  
黄卫  罗玉萍 《生物技术》2002,12(1):34-36
DNA分子标记是最为理想的遗传标记 ,依其多态性检出所用的分子生物学技术 ,大致可分为Southem杂交技术为核心的分子标记和PCR技术为核心的分子标记。前者的代表性技术有RELP(restrictionfragmentlengthpolymorphism)和DNA指纹技术 (DNAfingerprintingtechniques)。后者的代表性技术有RAPD(randomamplifiedpolymorphicDNA)、SCAR(sequencecharac teristicamplifiedregion)…  相似文献   

5.
利用 RFLP、SSR.AFLP和RAPD 4种分子标记方法研究了 15个玉米(Zea mays L.)自交系的遗传多样性,同时对4种标记系统进行比较。在供试材料中筛选到具多态性的RFLP探针酶组合56个,66对SSR引物,20个RAPD引物和9个AFLP引物组合,分别检测到多态性带167、201、87和108条。SSR标记位点的平均多态性信息量(PIC)最大(0.54),AFLP标记位点最小(0.36),但AFLP标记具有最高的多态性检测效率(Ai,32.2)。4种分子标记所得遗传相似系数相关性显著,比较相关系数表明 RAPD可靠性较低。依据 4种分子标记结果将 15个供试自交系划分为塘四平头、旅大红骨、兰卡斯特、瑞德和PN共5个类群,与系谱分析基本一致。认为SSR和RFLP两种分子标记方法适合进行玉米种质遗传多样性的研究。  相似文献   

6.
袁力行 Warbu.  M 《遗传学报》2000,27(8):725-733
利用RFLP、SSR、AFLP和RAPD4种分子标记方法研究了15个玉米(Zea mays L.)自交系的遗传多样性,同时对4种标记系统进行比较。在供试材料中筛选到具多态性的RFLP探针酶组合56个,676对SSR引物,20个RAPD引物和9个AFLP引物组合,分别检测到多态性带167、201、87和108条。SSR标记位点的平均多态性检测效率(Ai,32.2)。4种分子标记所得遗传相似自交系划分  相似文献   

7.
与西瓜野生种质抗枯萎病基因连锁的RAPD标记   总被引:23,自引:0,他引:23  
运用RAPD技术,采用混合分组分析(bulkedsegregantanalysis,BSA)方法进行了西瓜(Citrulluslanatus(Thunb.)Mansfeldvar. citroides) 野生种质PI296341 抗枯萎病基因连锁的分子标记研究。研究结果表明:西瓜野生种质P1296341 抗枯萎病生理小种1 的抗性由单显性基因控制,RAPD标记OPPOL/700 与其抗病基因连锁,其遗传距离为30 cM(centimorgan)。这为进行抗病分子标记辅助选择,以及最终定位与克隆其抗病基因打下了良好基础。  相似文献   

8.
大鼠RAPD标记的观察   总被引:1,自引:0,他引:1  
李昕权  李丰益 《遗传》1999,(1):8-10
采用随机扩增多态DNA(RAPD)技术,分析SD和Wistar二种大鼠的基因多态性,探讨用RAPD标记鉴别二种大鼠及其血标本实验中的认证,结果表明,二种大鼠表现出了各自不同的多态性RAPD标记,作为大鼠的分子标记,可在基因水平区别二种大鼠,故认为是一种大鼠研究的分子依据。  相似文献   

9.
葡萄感霜霉病基因RAPD标记的序列分析   总被引:1,自引:0,他引:1  
利用Wizard DNA clean-up system纯化葡萄感霜霉病基因RAPD遗传标记的DNA片段,用细菌质粒pGEM T-easy vector克隆该片段,采用自动荧光DNA测序仪对片段的核苷酸组成进行双向测序。来自欧洲葡萄粉红玫瑰的葡萄感霜霉病基因RAPD标记由835对核苷酸及其特定序列组成。所获的感霜霉病基因RAPD标记可以作为合成探针的基础,用于葡萄抗病育种过程中的早期选择及品种对霜  相似文献   

10.
显微分离出黑麦(SecalecerealeL.)1R染色体,用CohesiveadapterssingleprimerPCR(CASPPCR)方法进行体外扩增,以DIG11dUTP标记扩增产物为探针,进行Southern分子杂交,结果表明扩增产物来自黑麦1R染色体。用1/10体积的连接物转化E.coliDH5α,获得10000多个重组菌落。经酶切分析,克隆子的插入片段为250~500bp,为进一步筛选1R染色体的分子标记打下了基础  相似文献   

11.
In India, molecular mapping and tagging of agronomically important genes using RFLP and RAPD markers have been carried out in three different crops: rice, mustard and chickpea. In rice, tagging of genes for resistance to gall midge and blast has been accomplished. Molecular mapping of cooking quality traits in rice is in progress. For fingerpringting rice cultivars, suitable probe enzyme combinations have been identified. In mustard, a partial RFLP linkage map has been constructed and one of the yellow seed-coat colour loci has been mapped. Significant associations of RFLP markers with quantitative traits have also been established. Potential use of RAPD markers to identify heterotic groups among mustard accessions has been demonstrated. In chickpea, the occurrence of considerable interspecific DNA polymorphism as revealed by RAPD analysis has facilitated construction of a partial linkage map.  相似文献   

12.
Hordeum chilense is a potential source of useful genes for wheat breeding. The use of this wild species to increase genetic variation in wheat will be greatly facilitated by marker-assisted introgression. In recent years, the search for the most suitable DNA marker system for tagging H. chilense genomic regions in a wheat background has lead to the development of RAPD and SCAR markers for this species. RAPDs represent an easy way of quickly generating suitable introgression markers, but their use is limited in heterogeneous wheat genetic backgrounds. SCARs are more specific assays, suitable for automatation or multiplexing. Direct sequencing of RAPD products is a cost-effective approach that reduces labour and costs for SCAR development. The use of SSR and STS primers originally developed for wheat and barley are additional sources of genetic markers. Practical applications of the different marker approaches for obtaining derived introgression products are described.  相似文献   

13.
Summary Sequence characterized amplified regions (SCARs) were derived from eight random amplified polymorphic DNA (RAPD) markers linked to disease resistance genes in lettuce. SCARs are PCR-based markers that represent single, genetically defined loci that are identified by PCR amplification of genomic DNA with pairs of specific oligonucleotide primers; they may contain high-copy, dispersed genomic sequences within the amplified region. Amplified RAPD products were cloned and sequenced. The sequence was used to design 24-mer oligonucleotide primers for each end. All pairs of SCAR primers resulted in the amplification of single major bands the same size as the RAPD fragment cloned. Polymorphism was either retained as the presence or absence of amplification of the band or appeared as length polymorphisms that converted dominant RAPD loci into codominant SCAR markers. This study provided information on the molecular basis of RAPD markers. The amplified fragment contained no obvious repeated sequences beyond the primer sequence. Five out of eight pairs of SCAR primers amplified an alternate allele from both parents of the mapping population; therefore, the original RAPD polymorphism was likely due to mismatch at the primer sites.  相似文献   

14.
Gene mapping for a Cupressus species is presented for the first time. Two linkage maps for the Mediterranean cypress (Cupressus sempervirens) varieties, C. sempervirens var. horizontalis and C. sempervirens var. pyramidalis, were constructed following the pseudo-testcross mapping strategy and employing RAPD, SCAR and morphological markers. A total of 427 loci (425 RAPDs, two SCARs) representing parents and F(1) progeny were screened for polymorphism with 32 random decamer and two SCAR primers. A morphological marker defined as "crown form" was also included. Of 274 polymorphic loci, the 188 that presented Mendelian inheritance formed the mapping dataset. Of these loci, 30% were mapped into seven linkage groups for the horizontalis (maternal) and four linkage groups for the pyramidalis (paternal) map. The putative "crown form" locus was included in a linkage group of both maps. The horizontalis and the pyramidalis maps covered 160.1 and 144.5 cM, respectively, while genome length was estimated to be 1696 cM for the former variety and 1373 cM for the latter. The four RAPD markers most tightly linked to crown form were cloned and converted to SCARs. Each of the cloned RAPD markers yielded two to three different sequences behaving as co-migrating fragments. Two SCAR markers, SC-D05(432) and SC-D09(667), produced amplified bands of the expected sizes and maintained linkage with the appropriate phenotype, but to a lesser extent compared to their original RAPD counterparts. These linkage maps represent a first step towards the localization of QTLs and genes controlling crown form and other polygenic traits in cypress.  相似文献   

15.
RAPD markers generated by mixtures of two different primers were developed for octoploid × Tritordeum (amphiploid Hordeum chilense × Triticum aestivum) and its parents. Addition lines were used to identify 21 specific RAPD markers for the H. chilense chromosomes detectable in a wheat background. Ten RAPD bands were selected and eight of them were converted into dominant SCAR markers by direct sequencing of the RAPD products, avoiding the costly and time-consuming cloning step. The methodology overcomes some of the pitfalls associated with the election of the right clones when developing SCARs from RAPD markers. The SCARs generated have maintained both the chromosome specificity and the possibility of detection in a wheat background. This strategy provides a rapid method for the characterization of RAPD markers and for the development of PCR-based markers for both the characterization of the introgression of H. chilense in bread and durum wheat, as well as the efficient and reliable screening of tritordeum lines. This revised version was published online in June 2006 with corrections to the Cover Date.  相似文献   

16.
Two quantitative trait loci (QTLs), (QTLAR1 and QTLAR2) associated with resistance to ascochyta blight, caused by Ascochyta rabiei, have been identified in a recombinant inbred line population derived from a cross of kabuli×desi chickpea. The population was evaluated in two cropping seasons under field conditions and the QTLs were found to be located in two different linkage groups (LG4a and LG4b). LG4b was saturated with RAPD markers and four of them associated with resistance were sequenced to give sequence characterized amplified regions (SCARs) that segregated with QTLAR2. This QTL explained 21% of the total phenotypic variation. However, QTLAR1, located in LG4a, explained around 34% of the total phenotypic variation in reaction to ascochyta blight when scored in the second cropping season. This LG4a region only includes a few markers, the flower colour locus (B/b), STMS GAA47, a RAPD marker and an inter-simple-sequence-repeat and corresponds with a previously reported QTL. From the four SCARs tagging QTLAR2, SCAR (SCY17590) was co-dominant, and the other three were dominant. All SCARs segregated in a 1:1 (presence:absence) ratio and the scoring co-segregated with their respective RAPD markers. QTLAR2 on LG4b was mapped in a highly saturated genomic region covering a genetic distance of 0.8 cM with a cluster of nine markers (three SCARs, two sequence-tagged microsatellite sites (STMS) and four RAPDs). Two of the four SCARs showed significant alignment with genes or proteins related to disease resistance in other species and one of them (SCK13603) was sited in the highly saturated region linked to QTLAR2. STMS TA72 and TA146 located in LG4b were described in previous maps where QTL for blight resistance were also localized in both inter and intraspecific crosses. These findings may improve the precision of molecular breeding for QTLAR2 as they will allow the choice of as much polymorphism as possible in any population and could be the starting point for finding a candidate resistant gene for ascochyta blight resistance in chickpea.  相似文献   

17.
Two resistances to downy mildew derived from Lactuca serriola were characterized genetically and mapped using molecular markers. Classical genetic analysis suggested monogenic inheritance; however, the presence of multiple, tightly-linked genes in each case could not be eliminated. Therefore, they were designated resistance factors R17 and R18. Analysis with molecular markers known to be linked to clusters of resistance genes quickly revealed linkage of R18 to the major cluster of resistance genes and provided six linked markers, three RAPD (Random Amplified Polymorphic DNA) markers and three codominant SCAR (Sequence Characterized Amplified Region) markers. The mapping of R17 required the screening of arbitrary RAPD markers using bulked segregant analysis; this provided five linked markers, three of which segregated in the basic mapping population. This demonstrated loose linkage to a second cluster of resistance genes and provided additional linked markers. Two RAPD markers linked to R17 were converted into SCARs. The identification of reliable PCR-based markers flanking each gene will aid in selection and in combining these resistance genes with others.  相似文献   

18.
The randomly amplified polymorphic DNA (RAPD) method was used to investigate the genetic diversity in Xanthomonas cynarae, which causes bacterial bract spot disease of artichoke. This RAPD analysis was also intended to identify molecular markers characteristic of this species, in order to develop PCR-based markers which can be used to detect this pathogenic bacterium in artichoke fields. Among the 340 RAPD primers tested, 40 were selected on their ability to produce reproducible and reliable fingerprints in our genetic background. These 40 primers produced almost similar patterns for the 37 X. cynarae strains studied, different from the fingerprints obtained for other Xanthomonas species and other xanthomonad-like bacteria isolated from artichoke leaves. Therefore, X. cynarae strains form a homogeneous genetic group. However, a little DNA polymorphism within this species was observed and the collection of X. cynarae isolates was divided into two groups (one containing three strains, the second one including all other strains). Out of seven RAPD markers characteristic of X. cynarae that were cloned, four did not hybridize to the genomic DNA of strains belonging to other Xanthomonas species. These four RAPD markers were converted into PCR markers (specific characterized amplified regions [SCARs]); they were sequenced, and a PCR primer pair was designed for each of them. Three derived SCARs are good candidates to develop PCR-based tests to detect X. cynarae in artichoke fields.  相似文献   

19.
A set of 14 SCAR markers were developed starting from RAPD, AFLP and SAMPL analysis of several olive germplasm accessions. Eight RAPD, two AFLP and four SAMPL fragments were converted into dominant and codominant SCARs by cloning and sequencing the selected fragments. The markers obtained were evaluated on forty different olive cultivars from different Italian production areas (mainly from Liguria). The combined use of these SCARs made possible to univocally identify 26 cultivars while the remaining 14 will require the development of further markers since most of them are placed in a main group containing six genetically similar cultivars (among which Frantoio and Taggiasca) and four minor groups containing two cultivars each. A total of 31 different haplotypes were identified and the analysis of several individual plants indicated no intra-cultivar variability. Considering the SCAR polymorphism two alleles were scored for each markers with the only exception of markers IGPS3 and IGPS4 showing 4 alleles with 7 recognised groups and 5 alleles with 4 groups, respectively. Though less polymorphic in comparison with other markers like SSRs, the developed SCARs proved useful in genotype identification. In addition, they could potentially be used for breeding applications and forensic analysis. An erratum to this article is available at .  相似文献   

20.
RAPD技术是在PCR基础上发展起来的一种DNA多态性检测技术,已广泛应用于基因组研究的各个领域。本文概述了RAPD反应的原理、特点,总结了其在遗传多样性检测、亲缘关系鉴定、遗传连锁分析和数量性状的辅助标记选择等方面的应用,并肯定了RAPD在动物遗传育种领域的应用前景。  相似文献   

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