RFLP,RAPD,AFLP分子标记及其在植物生物技术中的应用

分子标记的创新、发展与应用对植物分子生物学和植物生长技术起了积极的推动作用。本文就ＲＦＬＰ、ＲＡＰＤ、ＡＦＬＰ分子标记及其在植物生物技术中应用作一介绍。     

生化标记和分子标记在蕈菌研究中的应用现状

本文介绍了目前在蕈菌研究中的酯酶同工酶标记,RAPD标记,RFLP标记,AFLP标记,简单重复序列标记和电泳核型等分子标记和生化标记在蕈菌遗传育种、菌株鉴定、遗传多样性研究、亲缘关系和基因定位等方面的研究、应用现状,包括原理、应用领域及最新研究进展。     

植物种群研究中的分子标记及其应用

综述了植物种群分子生态研究中各种标记法及其应用,包括等位酶、ＡＦＬＰ、ＲＡＰＤ、ＳＳＲ及ＲＦＬＰ等方法,并根据所涉及的有关研究论述及研究工作总结比较了各种方法应用在植物种群学研究中的利弊,强调应依不同的研究目的采用不同的分子标记方法,各种标记方法对解决不同的问题在质和量上是不同的。     

多个遗传标记情形下的RFLP的可诊断率估计

限制性酶切片段长度多态性(RFLP)作为共显性的遗传标记,已广泛应用于遗传病的产前诊断。为评价各遗传标记的适用性,本文给出了在多个遗传标记情形下的RFLP的可诊断率的估计方法,即在使用若干个遗传标记时,群体中可被诊断后代罹病与否的婚配类型的比例的估计方法,包括致病基因为常染色体显性、常染色体隐性、X连锁显性和X连锁隐性的情况,并认为增加遗传标记的个数和选择具较多等位基因的遗传标记,是提高产前诊断可诊断率的有效途径。同时,根据各遗传标记在群体中的多态性分布,可估计各遗传标记及其各种不同组合的可诊断率,以此选择在该群体中最为适合的遗传标记或其组合,以指导RFLP在产前诊断中的应用。     

小麦分子标记研究进展

一、小麦ＲＦＬＰ标记遗传图谱遗传连锁图既是遗传学本身的重要研究内容,又是育种等许多应用研究的理论依据和基础。数十年来,小麦遗传学家利用形态标记、生化标记和传统的细胞遗传学方法,为构建小麦遗传图谱进行了大量工作,并取得了一定的进展。但是由于形态标记和生化标记数目少,特殊遗传材料培育困难及细胞学工作量大,因而在ＲＦＬＰ遗传连锁图绘制之前,一直没有完整的小麦连锁图绘制出来。极大地限制了小麦遗传学理论研究与应用研究的进展。进入八十年代以来,分子标记的迅速发展大大促进了遗传连锁图的构建。１９８９年７月由美…     

分子标记及其在生态学中的应用

自 6 0年代发现等位酶 (allozyme)标记 ,70年代迅速发展 ;80年代出现DNA标记技术 ,至今仍是方兴未艾 ,形成了包括RFLP (restrictionfragmentlengthpolymorphism)、VNTR(variablenumbertendomrepeats)、RAPD (randomamplifiedpolymorphicDNA)在内的一系列分子标记技术。为解决生态学的个体识别、亲缘关系鉴定及种群间的遗传分化等问题提供了有力的武器。分子标记技术与生态学的结合 ,诞生了一门全新学科———分子生态学。     

水稻SSR标记的遗传多样性研究进展

本文从SSR标记优点和适用于研究水稻遗传多样性入手,综述了SSR标记在水稻核心种质构建与评价、遗传结构、稻种起源演化等方面的研究进展。总结了水稻遗传多样性的地带性特征（云南是中国稻种资源的最大遗传多样性中心和优异种质的富集地;西南稻区粳稻品种遗传多样性最丰富;南方稻区粳稻品种的遗传多样性高于北方粳稻遗传多样性）、遗传多样性与生态地理位置密切相关、目前水稻品种遗传基础狭窄、多样性降低等特征,分析了遗传多样性成因及影响因素,特别指出了育种行为对遗传多样性的影响,并针对当前水稻品种遗传多样性较低的问题提出了对策。     

RAPD分子标记及其在作物遗传育种中的应用

ＲＡＰＤ分子标记是一种新型的分了遗传标记,其原理是采用１０个碱基的随机引物,以基因组ＤＮＡ为模板进行ＰＣＲ随机扩增。其优越性在于其方法简单;分析中的花费少,用时短;分析所需的ＤＮＡ用量也不多等。本文着重介绍以电泳技术和ＰＣＲ扩增技术为核心的分子标记以及在农作物遗传育种中的应用。     

两个雌核发育白鲢群体同工酶分析遗传标记的确定

取源于武汉两个不同渔场两尾白鲢的卵子,经紫外照遗传物质失活的鲤鱼刺激雌核发育和热休克诱导第二极体保留的基因组操作技术,获得了两个不同的人工雌核发育白鲢群体。采用聚丙烯酰胺垂直板电泳技术,分析了这两个澡同人工雌核发育白鲢群体（分别称为Ｈｙ－Ｇ１和Ｈｙ－Ｇ２）内不同个体的肝脏、肌肉组织以及红细胞中脱氢酶（ＬＤＨ）、要酸氢酶（ＭＤＨ０、酯酶（ＥＳＴ）、超氧化物歧化酶（ＳＯＤ）等几种同工酶的表达谱式、并与     

AFLP标记在果树遗传育种研究中的应用

1993年 ,Ｚａｂｅａｕ等发明了一项专利技术 ,扩增片段长度多态性 (ａｍｐｌｉｆｉｅｄｆｒａｇｍｅｎｔｌｅｎｇｔｈｐｏｌｙｍｏｒｐｈｉｓｍ ,ＡＦＬＰ) ,该技术结合了ＲＦＬＰ技术和ＰＣＲ技术的优点 ,以其高效性和高重复性等特点 ,被广泛应用于多种植物的遗传育种研究。1 .ＡＦＬＰ技术的原理和特点ＡＦＬＰ技术是基于对限制性片段的选择性扩增 ,基因组ＤＮＡ被限制性内切酶切割后 ,将限制性片段末端连上双链接头 ,根据接头序列和相邻的限制性位点序列设计引物对限制性片段进行扩增。扩增产物经电泳检测后再比较其谱带的差异。ＡＦＬＰ…     

Comparison of PCR-based molecular marker analyses of Musa breeding populations

Progress in the breeding of plantain and banana has been restricted by the complex genetic structure and behaviour of cultivated polyploid Musa. Genetic improvement has been hindered due to the large amount of space required for growth and maintenance of plant populations, in addition to the long growth cycle and the low levels of fertility and seed viability characteristic of cultivated genotypes. Molecular marker assisted breeding has the potential to dramatically enhance the pace and efficiency of genetic improvement in Musa. This study was conducted to compare different PCR-based marker systems (RAPD, VNTR and AFLP) for the analysis of breeding populations generated from two diverse Musa breeding schemes. All three assays detected a high level of polymorphism between parental genotypes and within progeny populations. As expected, AFLP assays had by far the highest multiplex ratio while VNTR analysis detected the highest levels of polymorphism. AFLP analysis of a full-sib tetraploid hybrid population confirmed previous reports based on VNTR analysis, of a high frequency of recombination during 2n (3x) gamete formation by a triploid plantain landrace. In addition, both VNTR and RAPD analyses of a full-sib triploid hybrid population suggested a high frequency of homoeologous recombination during n (2x) gamete formation by tetraploid hybrids. In general, there was a poor correlation between estimates of genetic similarity based on different types of marker. The implications of these findings for the molecular breeding of Musa crops are discussed. This revised version was published online in June 2006 with corrections to the Cover Date.     

中国兰花资源分子标记鉴定研究进展

系统阐述了RAPD、SSR、RFLP、AFLP和其它几类分子标记在中国兰花种质资源鉴定及遗传多样性研究中的应用进展。同时指出,应用分子标记研究兰属植物的遗传多样性对兰属的科学分类及新品种选育等具有指导意义,并提出展望。     

Comparison of PCR-based marker systems for the analysis of genetic relationships in cultivated potato

The application of AFLPs, RAPDs and SSRs to examine genetic relationships in the primary northwestern European cultivated potato gene pool was investigated. Sixteen potato cultivars were genotyped using five AFLP primer combinations, 14 RAPD primers, and 17 database-derived SSR primer pairs. All three approaches successfully discriminated between the 16 cultivars using a minimum of one assay. Similarity matrices produced for each marker type on the basis of Nei and Li coefficients showed low correlations when compared with different statistical tests. Dendrograms were produced from these data for each marker system. The usefulness of each system was examined in terms of number of loci revealed (effective multiplex ratio, or EMR) and the amount of polymorphism detected (diversity index, or DI). AFLPs had the highest EMR, and SSRs the highest DI. A single parameter, marker index (MI), which is the product of DI and EMR, was used to evaluate the overall utility of each marker system. The use of these PCR-based marker systems in potato improvement and statutory applications is discussed.Abbreviations: PCR, polymerase chain reaction; AFLP, amplified fragment length polymorphism; RAPD, randomly amplified polymorphic DNA; DNA, deoxyribonucleic acid; EMR, effective multiplex ratio; DI, diversity index; MI, marker index; RFLP, restriction fragment length polymorphism.     

Comparing AFLP, RAPD and RFLP markers for measuring genetic diversity in melon

Three different types of molecular markers, RAPD, AFLP and RFLP were used to measure genetic diversity among six genotypes of Cucumis melo L. Each line represented a different melon genotype: Piel de Sapo, Ogen, PI161375, PI414723, Agrestis and C105. A number of polymorphic RAPD, AFLP and RFLP bands were scored on all materials and the genetic similarity measured. Clustering analysis performed with the three types of markers separated the genotypes into two main groups: (1) the sweet type, cultivated melons and (2) the exotic type, not cultivated melons. While the data obtained suggest that all three types of markers are equally informative, AFLPs showed the highest efficiency in detecting polymorphism. Received: 30 December 1999 / Accepted: 24 January 2000     

Conversion of an RAPD marker to an STS marker for barley variety identification

Barley (Hordeum vulgare L.) variety identification is important to the malting and brewing industries. Because many new malting cultivars (varieties) are closely related, new and more effective identification techniques are needed. We report on a series of techniques used to convert an RAPD marker to a more stable STS marker that can identify barley Stander from Robust, an important distinction for the American malting and brewing industries. The techniques included DNA extraction, RAPD amplification, random cloning of all amplified fragments, selection of clones by insert size, DNA sequencing of select inserts, design of a barley-based primer pair, and detection of a single nucleotide polymorphism using restriction endonucleaseAlu I. The barley-based primer pair was used to further sequence the RAPD fragment. Five single nucleotide polymorphisms between Robust and Stander exist, one of which was detected by electrophoresing DNA fragments differentially restricted byAlu I. The conversion technique was different from ones previously reported in that it did not require manual extraction of DNA fragments from a gel. This could be applied to other situations in which RAPD marker conversion would be desirable.     

RFLP、RAPD、AFLP在水稻农垦58S和1514中多态性比较

本文用ＲＦＬＰ、ＲＡＰＤ和ＡＦＬＰ三种分子标记技术对农垦５８ＳＸ１５１４组合及其Ｆ２极性集团进行了分析,比较了它们多成性和阳性的比率,结果显示,三种分子标记的多态性和与目的基因连锁的阳性比率分别为１９．９３％,５．２３％;１１．１７％,０．７６％和８６．４７％,７．５２％。ＡＦＬＰ的多态性比率和阳性比率均为最高。分析探讨了三种分子标记技术的优缺点及其在区间高分辩率作图和筛选与目的基因连锁标记中的运     

Estimating genetic drift and effective population size from temporal shifts in dominant gene marker frequencies

Measurement of temporal change in allele frequencies represents an indirect method for estimating the genetically effective size of populations. When allele frequencies are estimated for gene markers that display dominant gene expression, such as, e.g. random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) markers, the estimates can be seriously biased. We quantify bias for previous allele frequency estimators and present a new expression that is generally less biased and provides a more precise assessment of temporal allele frequency change. We further develop an estimator for effective population size that is appropriate when dealing with dominant gene markers. Comparison with estimates based on codominantly expressed genes, such as allozymes or microsatellites, indicates that about twice as many loci or sampled individuals are required when using dominant markers to achieve the same precision.