应用微卫星标记研究西藏野生大麦的遗传多样性

以西藏不同地区的106份野生大麦为材料,其中包括50份野生二棱大麦（HS）,27份野生瓶形大麦（HL）和29份野生六棱大麦（HA）,用Liu等（1996）发表的SSR连锁图的每个连锁群的两个臂的不同位置上选取3～5个共30个SSR标记,研究了西藏3类野生大麦的遗传多样性。结果表明,这3类野生大麦在遗传组成及等位变异频率分布上存在着明显的遗传分化。在总样本中,共检测到229个等位变异,平均每个SSR位点检测到7．6个等位变异,其中70个为这3类野生大麦间共同的等位变异,等位变异数在这3类野生大麦间有明显的差异,亚种问的遗传多样性明显高于亚种内的遗传多样性。其遗传多样性大小顺序为HS〉HL〉HA。聚类分析表明,野生二棱大麦、野生六棱大麦分别聚在不同的两类,而野生瓶形大麦中各有约50％的材料分别聚在这两类。根据本研究及前人研究结果,我们认为中国栽培大麦是从野生二棱大麦经野生瓶形大麦向野生六棱大麦进化的。该结果支持了栽培大麦起源的“野生二棱大麦单系起源论”的观点。     

微卫星DNA标记技术及其在植物研究中的应用

微卫星DNA(或简单重复序列,simplesequencerepeats,SSR)是继RFLP、RAPD等分子标记后出现的第二代分子标记技术。随着分子生物学的发展,微卫星标记技术在植物基因组中的应用越来越广泛。由于SSR具有多态性高,呈共显性遗传,遵守孟德尔式分离,在数量上没有生物学的限制,实验操作简单,对样品质量要求不高等特点,因此被广泛用于植物遗传图谱的构建、基因定位、构建指纹图谱、遗传多样性及物种进化与亲缘关系的研究等方面。     

微卫星DNA标记开发技术进展及其在经济植物研究中的应用

微卫星分子标记技术被广泛应用于分子生物学研究中,具有多态性高、重复性好、共显性表达、杂合度高等特点,在种群遗传多样性、遗传图谱构建等领域发挥不可替代的作用。近年来,随着新一代高通量测序技术的进一步成熟,简单重复序列(simple sequence repeat,SSR)标记的开发技术与应用得到了进一步的发展。现主要对基于高通量测序的微卫星分子标记最新开发技术进行介绍,并从遗传多样性分析、遗传图谱构建、品种鉴定及分子辅助育种等方面,总结近年来SSR标记技术在经济植物研究中的最新应用,最后对SSR技术的应用前景进行展望,以期为利用微卫星技术进行经济植物研究提供参考。     

微卫星DNA标记技术及其在昆虫学上的应用

微卫星DNA标记作为一种多态性和稳定性高、重复性好、呈共显性的分子遗传标记技术,目前已被广泛应用于昆虫学的研究中。本文介绍了微卫星DNA标记的基本原理和特点,并综述了近年来该技术在昆虫种群遗传结构及分化、生物学特性与习性、遗传图谱的构建、基因定位以及系统发生等领域中的应用。     

应用SSR标记对61个国家大麦遗传多样性的研究

使用全自动基因分析仪（ABI-3700 DNA Analyzer）,用35对SSR荧光标记引物对来自61个国家的2625份大麦种质资源的遗传多样性进行了分析,得出如下结果：（1）35对引物在2625份大麦种质资源中共检测到2063个等位变异,每个位点的等位变异数为30～79个,平均为58.94;多样性指数为1.60～3.66,平均为2.80;（2）在大麦的7条染色体中,每条染色体的等位变异数不等,染色体等位变异数从多到少排序为5、7、2、6、3、4、1,遗传多样性指数从高到低依次为2、6、7、3、5、4、1,综合这两项指标,第2、7条染色体遗传多样性较高,第1条染色体的遗传多样性最低;（3）约旦、伊朗、叙利亚和利比亚等国家大麦的遗传多样性指数最高,分别为2.58、2.53、2.50和2.38;而埃塞俄比亚、叙利亚、伊朗和约旦等国家的大麦资源则具有较多的等位变异,分别为29.03、25.57、25.11和24.60,综合分析,约旦、伊朗、叙利亚和土耳其大麦遗传多样性较高。     

花生微卫星标记的研究进展

近年来花生微卫星标记的开发取得了一定的进展,初步揭示了花生在DNA水平上的遗传多样性。花生微卫星标记的开发途径主要包括通过构建小片段基因组文库开发基因组SSR标记,根据花生EST序列开发EST-SSR标记,根据豆科植物序列信息和SSR标记开发花生SSR标记,将SSR标记与其它分子标记结合开发新的DNA标记,以及基于SSR核心序列开发ISSR标记。花生微卫星标记主要应用于遗传多样性研究、遗传图谱与品种指纹图谱构建以及分子标记辅助育种等领域。本文综述了花生SSR标记开发研究的进展及应用。     

花生微卫星标记的研究进展

近年来花生微卫星标记的开发取得了一定的进展, 初步揭示了花生在DNA水平上的遗传多样性。花生微卫星标记的开发途径主要包括通过构建小片段基因组文库开发基因组SSR标记, 根据花生EST序列开发EST-SSR标记, 根据豆科植物序 列信息和SSR标记开发花生SSR标记, 将SSR标记与其它分子标记结合开发新的DNA标记, 以及基于SSR核心序列开发ISSR标记。花生微卫星标记主要应用于遗传多样性研究、遗传图谱与品种指纹图谱构建以及分子标记辅助育种等领域。本文综述了花生SSR标记开发研究的进展及应用。     

基于SSR分子标记的地被菊遗传多样性分析和指纹图谱构建

为了从分子水平上研究地被菊(Chrysanthemum×morifolium)种质资源的遗传多样性并建立地被菊品种的指纹图谱库,筛选出多态性高的引物用于地被菊品种间鉴定、亲缘关系分析和分子标记辅助选种体系的建立,本研究利用多态性好、条带清晰、重复性好的12对引物对91份地被菊材料和14份菊属近缘种材料进行简单重复序列(simple sequence repeat,SSR)分子标记和遗传多样性分析,从12对引物中筛选出9对核心引物对受试材料进行指纹图谱构建。结果显示,在105个样品中,12对引物检测出104个等位位点,范围为2−26,平均每个点位检测出9.25个等位基因数,平均每个点位检测得到的有效等位基因数(number of effective alleles,Ne)为2.7456,范围为1.2760−4.7425;Shannon信息指数(Shannon genetic diversity index,I)变化范围是0.5133−2.2399,均值是1.2090;Nei’s基因多样性指数(Nei’s gene diversity index,H)范围是0.2163−0.7891,均值是0.5780;观测杂合度(observed heterozygosity,Ho)的范围是0.2233−0.8952,均值是0.5575;期望杂合度(expected heterozygosity,He)的范围是0.2174−0.7933,均值是0.5808;多态信息含量(polymorphism information content,PIC)值变化范围是0.2115−0.7740,均值是0.5329;遗传相似性系数(genetic similarity,GS)范围为0.2285−1.0000,均值是0.6083。聚类分析表明,在遗传距离(genetic distance,GD)=0.30时,受试材料可以分为两个类群。Structure群体结构分析将受试材料分为3个种群和1个混合种群。从12对引物中筛选出可完全区分105份受试材料的9对核心引物,构建了91份地被菊材料和14份菊属近缘种材料的指纹图谱。地被菊材料之间具有显著的遗传差异和丰富的遗传多样性,对于地被菊的园林应用和品种选育具有重要意义。地被菊品种和菊属近缘种的指纹图谱库的构建,一定程度上揭示了105份实验材料的亲缘关系,为今后地被菊品种鉴定与筛选体系的研究提供了技术支撑。     

微卫星标记及其在鸟类中的应用

微卫星DNA作为一种分子标记,以其诸多优点被认为是各类遗传标记中最有价值的一种。目前已被广泛应用于鸟类的研究中。本文介绍了微卫星DNA标记的分布特点、突变机制和作为分子标记的特点,并综述了微卫星DNA标记在鸟类的亲权分析及种类鉴定、遗传图谱构建及基因定位、物种进化历史揭示和遗传多样性研究中应用现状。     

微卫星DNA在分子遗传标记研究中的应用

随着种群遗传学的发展 ,分子遗传标记特别是微卫星标记已经成为研究种群遗传的有力工具。本文就微卫星遗传标记的研究背景、技术应用以及优势与不足等方面进行了综述。     

小麦SSR标记的发展及应用

微卫星是以1～6个碱基为基本单元的串联重复序列,由于具有共显性、多态性高和容易用PCR方法检测等特点,是非常有用的遗传标记。在小麦中,SSR标记已广泛应用于遗传图谱的构建、遗传多样性、品种及基因型鉴定、目的基因,以及QTL的标记和标记辅助选择育种。 Abstract:Microsatellites are simple,tandemly repeated one to six nucleotide sequence motifs.They are very useful as genetic markers because they are co-dominant,detect high levels of allelic diversity,and are easily assayed by the polymerase chain reaction ( PCR ).In wheat,SSR markers have been applied to genetic mapping,detection of genetic diversity,identification of varieties and genotypes,gene tagging,QTL analysis,and marker-assisted selection.     

Barley microsatellites: allele variation and mapping

Microsatellites have developed into a powerful tool for mapping mammalian genomes and first reports about their use in plants have been published. A database search of 228 barley sequences from GenBank and EMBL was made to determine which simple sequence repeat (SSR) motif prevails in barley. Nearly all types of SSRs were found. The (A)n and (T)n SSRs occurred more often than (C)n and (G)n for n10. Among the dinucleotide repeats, the (CG)n SSRs occurred least often. Trinucleotide repeats did not occur with n>7 and there is no correlation between the GC content in the trinucleotide motifs and the number of observed SSRs. Analysing 15 different microsatellites with 11 barleys yielded 2.1 alleles per microsatellite. Sequencing 25 putative microsatellites showed that the resolution capacity of highquality agarose gels was sufficient to determine differences of only three base paris. Five microsatellites were mapped on three different chromosomes of a barley RFLP map.     

Microsatellite genotyping by primer extension and MALDI-ToF mass spectrometry

A novel method for genotyping microsatellite alleles using primer extensions and mass spectrometry analysis has been developed. Following PCR amplification of the target region, a genotyping primer, with its 3′ end directly flanking the microsatellite repeats, was extended by a mixture of dNTPs complementary to the nucleotides composing the microsatellite. The length and molecular weight of extended primers vary with the number of repeats present in the allele(s) under examination. The weights of extension products were determined using matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-ToF MS) and used to identify genotypes on the basis of differential primer extension. This is a platform that is not gel based and is amenable to multiplexing and automation. The technique enables identification of heterozygous progeny in which alleles differ by a single trinucleotide repeat. The method is illustrated by genotyping a polymorphic microsatellite identified in an intron of the barleyMlo gene.     

Microsatellite marker development,mapping and applications in rice genetics and breeding

Microsatellites are simple, tandemly repeated di- to tetra-nucleotide sequence motifs flanked by unique sequences. They are valuable as genetic markers because they are co-dominant, detect high levels of allelic diversity, and are easily and economically assayed by the polymerase chain reaction (PCR). Results from screening a rice genomic library suggest that there are an estimated 5700-10 000 microsatellites in rice, with the relative frequency of different repeats decreasing with increasing size of the motif. A map consisting of 120 microsatellite markers demonstrates that they are well distributed throughout the 12 chromosomes of rice. Five multiple copy primer sequences have been identified that could be mapped to independent chromosomal locations. The current level of genome coverage provided by these simple sequence length polymorphisms (SSLPs) in rice is sufficient to be useful for genotype identification, gene and quantitative trait locus (QTL) analysis, screening of large insert libraries, and marker-assisted selection in breeding. Studies of allelic diversity have documented up to 25 alleles at a single locus in cultivated rice germplasm and provide evidence that amplification in wild relatives of Oryza sativa is generally reliable. The availability of increasing numbers of mapped SSLP markers can be expected to complement existing RFLP and AFLP maps, increasing the power and resolution of genome analysis in rice.     

利用微卫星标记分析东平湖黄颡鱼的遗传多样性

采用27对鲤微卫星引物对山东东平湖黄颡鱼进行全基因组扫描,结果有19对引物能获得稳定的扩增条带,其中有6个微卫星位点具有多态性。对这6个位点的扩增产物进行分析,结果显示:6个位点共检测到22个等位基因,每个位点的等位基因数从2个到6个不等;平均基因纯合率为41.67%,平均多态信息含量(PIC)为0.488,平均杂合度为0.5833。这表明东平湖黄颡鱼种群结构合理,群体遗传多样性较丰富,种质资源处于安全状态。     

Characterization of Fusarium poae Microsatellite Markers on Strains from Switzerland and Other Countries

Fusarium poae is a pathogen of increasing importance within the disease complex Fusarium head blight (FHB). Eleven microsatellite markers were developed, and 72 F. poae strains from Switzerland and other countries were used to assess the level of marker polymorphism. The number of alleles for each of the markers ranged from 4 to 15, and the average gene diversity was 0.62, ranging from 0.25 to 0.84. Using these novel markers, 44 genotypes could be differentiated among all F. poae strains. Two genotypes were represented by nine and ten strains, respectively, deriving from distinct geographic areas within Switzerland and indicating a potential selection advantage. Four markers were F. poae‐specific, whereas seven markers also yielded amplification products in one to four strains of five other Fusarium species. Of the latter, five markers revealed F. poae‐specific allele size ranges. Hence, these microsatellite markers could be used both for FHB species differentiation and for intra‐specific distinction of F. poae strains.     

应用微卫星标记进行大豆种质多样性和遗传变异性分析

微卫星标记又称ＳＳＲ标记是近年来发展的一种新型的分子标记可有效地进行基因型鉴定,系统谱分析并可估算材料间的遗传距离。用５对ＳＳＲ引物对１５份大豆材料进行扩增,共得到２１条多态性条带。每个ＳＳＲ座位的等侠基因数目为３～＾个,基因多样性范围为０．４３７～０．６６８,对这些材料进行了遗传距离分析。家系分析表明微卫星ＤＮＡ经过多世代的九分裂的后产生了突变,在ＲＩＬ Ｆ８代中有的个体在个别ＳＳＲ等基因的大小     

Genetic Diversity and Natural Population Structure of Cacao (Theobroma cacao L.) from the Brazilian Amazon Evaluated by Microsatellite Markers

A sample of 94 accessions of Theobroma cacao L. (cacao), representing four populations from the Brazilian Amazon (Acre, Rondônia, lower Amazon and upper Amazon) were analyzed using microsatellite markers to assess the genetic diversity and the natural population structure. From the 19 microsatellite loci tested, 11 amplified scorable products, revealing a total of 49 alleles, including two monomorphic loci. The Brazilian upper Amazon population contained the largest genetic diversity, with the most polymorphic loci, the highest observed heterozygosity; and the majority of rare alleles, thereby this region might be considered part of the center of diversity of the species. The observed heterozygosity for all the Brazilian populations (H _o = 0.347) was comparable with values reported for other similar upper Amazon Forastero cacao populations, with the Acre and Rondônia displaying the lowest values. The lower Amazon population, traditionally defined as highly homozygous, presented an unexpectedly high observed heterozygosity (H _o = 0.372), disclosing rare and distinct alleles, with large identity with the upper Amazon population. It was hypothesized that part of the lower Amazon population might derive from successive natural or intentional introduction of planting material from other provenances, mainly upper Amazon. Most of the loci exhibited a lower observed heterozygosity than expected, suggesting that self-pollination might be more common than usually assumed in cacao, but excess of homozygotes might also derive from sub-grouping (Wahlund effect) or from sampling related individuals. Most of the gene diversity was found to occur within groups, with small differentiation between the four Brazilian Amazon populations, typical of species with high gene flow.