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1.
AFLP(扩增片段长度多态性)是检测DNA多态性的一种高效的分子标记技术,该技术的运用不需要预知基因组的序列特征,可适用于任何来源和各种复杂度的DNA,兼具RFLP技术的可靠性和RAPD技术的方便性。近年来,AFLP技术已在干果遗传育种研究中得到广泛应用。本文对AFLP技术的基本原理和反应程序进行了介绍,综述了其在干果种质资源研究、遗传图谱构建、基因标记、分子辅助选择、基因表达等方面的应用,并对AFLP技术存在的问题及其在干果遗传育种研究上的应用前景进行了探讨。  相似文献   

2.
分子标记技术在棉花品种鉴定上的研究进展   总被引:12,自引:5,他引:7  
 DNA指纹技术的发展大体分为三个阶段:第一代的分子标记是以Southern杂交为基础的RFLP,第二代分子标记是以PCR为基础的各种DNA指纹标记,第三代分子标记是以单核苷酸多态性为基础的SNP。本文概述了品种鉴定技术的研究与应用进展,重点介绍了用于棉花品种鉴定的四种主要的分子标记技术:RFLP、RAPD、AFLP和SSR,对每一种分子标记的技术原理、在棉花品种鉴定上的应用研究状况及存在的优缺点进行了具体阐述。通过比较分析,提出了SSR标记适用于棉花品种鉴定的独特优越性。并对基于SSR分子标记技术构建我国棉花品种DNA指纹库的重要意义与必要性进行了分析。  相似文献   

3.
一、引言DNA限制性片段长度多态性(Restriction Fragment Length Polymorphism简称RFLP)是近年来分子遗传学研究中发展起来的一种新技术。其技术要点是:DNA经限制性内切酶酶切后,产生不同长度的片段;经琼脂糖电泳分离这些不同的片段,印迹转移至硝酸纤维素滤膜上;然后用一放射性同位素标记的特定DNA克隆(探针)杂  相似文献   

4.
DNA分子标记及其在遗传育种上的应用   总被引:5,自引:0,他引:5  
杨文鹏 《种子》1997,(2):30-33
本文比较系统地介绍了DNA分子标记的种类(RFLP、“小卫星”DNA、“微卫星”DNA、AP-PCR、RAPD和AFLP等)、含义、技术、原理和特点,重点阐述TDNA分子标记在遗传育种上的应用前景。  相似文献   

5.
DNA分子标记是继形态标记、细胞标记和生化标记之后发展起来的一种新的较为理想的遗传标记,在甘蔗育种方面发挥了重要作用。DNA分子标记包括:限制性片段长度多态性(RFLP)、随机扩增多态性DNA(RAPD)、扩增片段长度多态性(AFLP)、简单重复序列(SSR)、染色体或基因组原位杂交(GISH)、荧光原位杂交(FISH)等。本文对植物DNA分子标记的原理、特点及其在甘蔗种属鉴定、遗传多样性分析、分子图谱构建及病害分子诊断等方面的研究进行简要的概述。  相似文献   

6.
简要介绍了5种传统的、最常用的DNA分子标记(RFLP、RAPD、AFLP、SSR和SNP)的技术原理及它们的优缺点,也总结了TRAP这种新产生的分子标记的技术原理、优点及应用前景.综述了这几类分子标记在花生种质进化、遗传多样性分析、分子图谱构建及抗虫、抗病等方面的研究.利用SSR和RAPD标记能够发现野生种和栽培种多态性进而实现分子标记对花生的遗传多样性分析,可以将许多花生品种分为不同的品种群,能够对花生进行种质进化研究.RFLP和AFLP技术利于花生图谱构建,利用DNA中特定的限制性酶切位点上碱基对的改变及酶切位点之间的分子重排,可以发现花生品种间的DNA许多多态性位点,进而绘制分子标记图谱.AFLP技术在花生青枯菌和花生抗黄曲霉的研究方面有很大进展.RAID技术在花生根瘤菌、花生线虫病等方面已有显著进展.最后对分子标记在花生育种中的应用前景进行了简单展望.  相似文献   

7.
植物根结线虫分子鉴定研究进展   总被引:12,自引:0,他引:12  
从RFLP、RAPD和SCAR、rDNA-ITS-PCR、AFLP等几方面概述了分子技术在植物根结线虫鉴定中的应用现状。分子技术用于检测植物根结线虫种间和种内群体间在DNA水平上的根本差异,为根结线虫的鉴定提供快速、准确的方法。  相似文献   

8.
DNA分子标记技术在农作物品种鉴定上的应用   总被引:2,自引:0,他引:2  
李志勇  谢华峰  张力  赵磊 《种子科技》2010,28(10):19-21
文章概述了利用DNA分子标记技术(RFLP、RAPD、SSR、AFLP、ISSR和STS等)进行品种鉴定的原理、特点和研究概况,从DNA分子水平上进行品种鉴定具有准确可靠、成本低、自动化、不受环境影响等优点,DNA分子标记技术在品种鉴定方面具有广阔的应用前景。  相似文献   

9.
分子标记技术在番茄抗性育种中的应用   总被引:2,自引:2,他引:0  
介绍了DNA分子标记的各种类型,RFLP、RAPD、SSR、AFLP和SNP标记是植物遗传作图最常用的,介绍了各种类型的使用范围以及优点和缺点。综述了近年来DNA分子标记技术在番茄抗性育种中的应用,包括耐冷性,耐盐性,抗病性,抗虫害等方面的应用,并就今后番茄分子育种主要研究方向进行了讨论。  相似文献   

10.
PAPD(随机扩增多态性DNA)是一项新兴的分子标记技术。该技术与其它分子标记技术如RFLP、DNA指纹图谱、SSCP等相比具有如下特点:1)对受试基因组无特定的要求,无需知道模板DNA序列的信息;2)不需要使用放射性标记;3)仅需毫微克数量级的DNA样品,一般1次扩增只需10一50ng的DNA;4)扩增引物随机设定,可从许多物种中检测出大量的多态DNA;5)检测灵敏度高,操作简单快捷,适合大量样本的快速分析。因而,RAPD技术正越来越广泛地应用于分子系统学研究、品种鉴定、基因组研究、遗传连锁图谱的构建、基因定位等研究领域。  相似文献   

11.
A DNA polymorphison assay, random amplified polymorphic DNA (RAPD), was developed in 1990 based on amplification by the polymerase chain reaction (PCR) of random DNA segments and single primers of arbitrary uncleotide sequnce. The major advantage of this assay, contrasting with the restriction fragment length polymorphism (RFLP), is that there is no requirement for any specific sequence information on the target genome, as well as rapidity and simplicity.It has been widespread applied in life science researches such as the creation of high density genetic mapping, molecular phylogenetic study and gene location.  相似文献   

12.
N. Stein    G. Herren  B. Keller   《Plant Breeding》2001,120(4):354-356
Gene mapping and marker‐assisted selection in complex, polyploid genomes still relies strongly on restriction fragment length polymorphism (RFLP) analysis, as conversion of RFLP to polymerase chain reaction (PCR) markers can be very difficult. DNA extraction in amounts suitable for RFLP analysis represents the most time‐consuming and labour‐intensive step in molecular marker analysis of plant populations. In this paper, a new flexible method for plant DNA extraction is presented. It allows a high‐throughput of samples in a short time without the need for freezing or lyophilizing the plant material. The method allows the isolation of genomic DNA with a yield of 100 μg for a minimal amount of 200 mg of leaf material. This is sufficient for work with large‐genome plant species such as hexaploid wheat, where 20 μg of genomic DNA are required for a single RFLP analysis.  相似文献   

13.
Summary A linkage map for watermelon (Citrullus lanatus) was constructed on the basis of RADP, ribosomal DNA restriction fragment length polymorphism (RFLP), isozyme, and morphological markers using F1BC1. A segregating population of 78 individuals was the result of a backcross of a cultivated inbred line (H-7; Citrullus lanatus; 2n=22) and a wild form (SA-1; C. lanatus; 2n=22), in which the latter was the recurrent (male) parent. A total of 69 RAPD, one RFLP, one isozyme, and three morphological markers was found to segregate in the BC1 population. Linkage analysis revealed that 62 loci could be mapped to 11 linkage groups that extended more than 524 centimorgans (cM), while 12 loci segregated independently of all other markers. The locus for exocarp color was linked to two RAPD markers within a region of 5 cM on linkage group 4. The locus for flesh color was linked to a RAPD marker within a region of 30 cM on linkage group 6. The isozyme marker GOT was located on the linkage group 1. Linkage group 2 contained a locus for ribosomal DNA within 5 cM of a RAPD marker. Half of the RAPD markers on the linkage group 7 displayed severely distorted segregation. The construction of linkage map using molecular markers is necessary for the breeding of watermelon to introduce useful gene of wild watermelon efficiently. However the linkage map that was constructed for the most part on the basis of RAPD markers could not cover significant parts of the genome, the linkage map provides breeders of watermelons the possibility of tagging useful agronomic traits, as well as the gene for exocarp color.Abbreviations RAPD random amplified polymorphic DNA - RFLP restriction fragment length polymorphism - GOT glutamate oxaloacetate transaminase - MDH malate dehydrogenase - ACP acid phosphatase - 6PGH 6-phosphogluconate dehydrogenase  相似文献   

14.
W. L. Wei    H. Z. Wang    G. H. Liu 《Plant Breeding》2009,128(4):426-428
It is very important for rapeseed hybrid production to develop and utilize a novel cytoplasmic male sterility (CMS) system concerning the possible risk because of a narrow cytoplasm background. Here the anatomy of anther development in the CMS system, named NCa , was observed using light microscope and transmission electron microscope, and the restriction fragment length polymorphism (RFLP) analyses of mitochondrial DNA of NCa sterile line were also performed in comparison with the other rapeseed sterile lines, such as pol , nap , ogura , and tour . The anther abortion of this CMS line occurred at the later uninucleate microspore stage, and the anatomic aborting characteristics were obviously different from all the other rapeseed CMS lines reported before. The RFLP analyses revealed that five probe/enzyme combinations could distinguish the five CMS lines. The results of anatomic observations and mitochondrial DNA polymorphism indicated that the NCa CMS system is a novel one which differs from the pol , nap , ogura , and tour systems.  相似文献   

15.
Cytoplasmic male sterile (CMS) plants were obtained by asymmetrical protoplast fusion between red cabbage (fertile) and normal radish (fertile). The CMS plants showed restriction fragment length polymorphism (RFLP) patterns of mitochondrial (mt) DNA that were different from those of both parental lines. PCR analysis of mtDNA of the CMS plants and though Southern blot analysis showed that the mtDNA of the CMS line had the characteristics of the ogura CMS type. The results suggested that the orf138 gene and the ogura type atp6 gene are present in normal fertile cabbage and radish at a low copy level. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

16.
A. Graner    S. Streng    A. Drescher    Y. Jin    I. Borovkova  B. J. Steffenson 《Plant Breeding》2000,119(5):389-392
Leaf rust of barley, caused by Puccinia hordei Otth, is an important foliar disease in most temperate regions of the world. Sixteen major leaf rust resistance (Rph) genes have been described from barley, but only a few have been mapped. The leaf rust resistance gene Rph7 was first described from the cultivar ‘Cebada Capa’ and has proven effective in Europe. Previously mapped restriction fragment length polymorphism (RFLP) markers have been used to determine the precise location of this gene in the barley genome. From the genetic analysis of a ‘Bow‐man’/‘Cebada Capa’ cross, Rph7 was mapped to the end of chromosome 3HS, 1.3 recombination units distal to the RFLP marker cMWG691. A codominant cleaved amplified polymorphic site (CAPS) marker was developed by exploiting allele‐specific sequence information of the cMWG691 site and adjacent fragments of genomic DNA. Based on the large amount of polymorphism present in this region, the CAPS marker may be useful for the marker‐assisted selection of Rph7 in most diverse genetic backgrounds.  相似文献   

17.
Y. C. Liao  R. E. Niks 《Euphytica》1991,53(2):115-119
Summary A set of 14 probes from wheat cDNA clones was used to search for restriction fragment length polymorphism (RFLP) in six barley lines. The degree of polymorphism among the lines varied greatly between probes and between the various restriction enzymes. Two probes revealed a high degree of polymorphism in all probe/enzyme combinations. Seven of 14 probes did not reveal RFLP. The average level of polymorphism based on all 840 pairwise comparisons was 14.0%, which is higher than has been reported in wheat, but lower than in maize, rice, potato and lettuce. Most of the probes that detected RFLP correspond to sites on the long arms of wheat chromosomes.  相似文献   

18.
Summary Cultivars of Cucurbita pepo and other Cucurbita species were characterized by RFLP analysis using different fragments of the ribosomal intergenic spacer (IGS) of Cucurbita pepo as hybridization probes. Several cultivars could be distinguished by a specific rDNA restriction pattern, whereas some cultivars showed an identical RFLP pattern suggesting a closer relationship. Other species of the genus Cucurbita exhibited strong cross-reaction with the C. pepo spacer probes, in contrast to DNA of Cucumis species which did not cross-hybridize.Abbreviations IGS intergenic spacer - ITS internal transcribed spacer - kbp kilo base pairs - rDNA ribosomal DNA - RFLP restriction fragment length polymorphism - rRNA ribosomal RNA  相似文献   

19.
Previously chromosome 3A of wheat (Triticum aestivum L.) was reported to carry genes influencing yield, yield components, plant height, and anthesis date. The objective of current study was to survey various molecular marker systems for their ability to detect polymorphism between wheat cultivars Cheyenne(CNN) and Wichita (WI), particularly for chromosome3A. Seventy-seven `sequence tagged site' (STS), 10simple sequence repeat (SSR), 40 randomly amplified polymorphic DNA (RAPD) markers, and 52 restriction fragment length polymorphism (RFLP) probes for wheat homoeologous group 3 chromosomes, were investigated. Three (3.9%) STS-PCR primer sets amplified polymorphic fragments for the two cultivars, of which one was polymorphic for chromosome 3A. Sixty percent of SSR markers detected polymorphism between CNN and WI of which 50% were polymorphic for chromosome 3A. Twenty percent of RAPD markers detected polymorphism between CNN and WI in general, but none of these detected polymorphism for chromosome 3A. Of the fifty-two RFLP probes, 78.8% detected polymorphism between CNN and WI for group 3 chromosomes with one or more of seven restriction enzymes and 42% of the polymorphic fragements were for chromosome 3A. These high levels of RFLP and SSR polymorphisms between two related wheat cultivars could be used to map and tag genes influencing important agronomic traits. It may also be important to reconsider RFLP as the most suitable marker system at least for anchor maps of closely related wheat cultivars. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

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