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A survey of the microsatellite content of barley, and several applications of microsatellite polymorphism.

机译：大麦中微卫星含量的调查以及微卫星多态性的几种应用。

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摘要

This thesis investigates the abundance and frequency of microsatellite DNA in barley (Hordeum vulgare L.) genomic database sequences, and demonstrates the versatility and limitation of barley microsatellite-based molecular markers.;Searches of the GenBank and EMBL databases yielded 217 perfect (uninterrupted) microsatellites, consisting of 44 dimeric, 65 trimeric, 73 tetrameric, 22 pentameric and 13 hexameric repeat containing regions. The majority of trimeric and hexameric microsatellites were found in protein coding regions, while all but five of the other microsatellites were located in introns or gene flanking sequences.;Microsatellite polymorphism successfully distinguished the cultivars in a Canadian representative set, using as few as two microsatellite-based primer sets. Twenty-six polymorphic loci were detected across this cultivar set, with two to five alleles found per polymorphic locus. Diversity index values ranged from 0.17--0.76. Correlation was found between allele number and size of dimeric microsatellites, particularly for AG/CT containing repeat regions. Interpretable data from microsatellite loci was equally easy to obtain using DNA extracted from leaf, seed or malt sources. Previously unpublished data and analysis were also reported from collaborative work on a set of six-row cultivars.;The stability of microsatellites over time was examined for 11 and five microsatellites across the pedigrees of the barley lines Harrington and WM872-3, respectively, and nine microsatellites across a set of 15 doubled haploid lines. No new microsatellite allele formation was detected within these lines for the tested microsatellites. During these experiments, within-cultivar microsatellite variation was detected in three cultivars for the compound dimeric microsatellite HVM40 and the perfect GA-containing microsatellite HVM68, thus illustrating the sensitivity of microsatellite markers, and indicating that intravarietal microsatellite variation should be measured before using them as molecular markers.;Nine primers designed to amplify barley AG/CT-containing microsatellites were used successfully to amplify sequences in hexaploid wheat, rye and oat cultivars. Polymorphism was detected across 12 wheat cultivars for two of the nine primer sets, providing preliminary evidence for microsatellite conservation across cereals.
机译：本文研究了大麦（Hordeum vulgare L.）基因组数据库序列中微卫星DNA的丰度和频率，并证明了基于大麦微卫星的分子标记的通用性和局限性。通过GenBank和EMBL数据库的搜索获得了217个完美（不间断）微卫星，由44个二聚体，65个三聚体，73个四聚体，22个五聚体和13个六聚体重复区域组成。大多数三聚体和六聚体微卫星存在于蛋白质编码区域，而除五个以外的其他所有微卫星都位于内含子或基因侧翼序列中。；微卫星多态性成功地区分了加拿大代表性组中的栽培品种，仅使用了两个微卫星引物组。在该品种组中检测到26个多态位点，每个多态位点发现2至5个等位基因。多样性指数值范围从0.17--0.76。发现等位基因数量和二聚体微卫星大小之间存在相关性，特别是对于含有AG / CT的重复区域。使用从叶片，种子或麦芽来源提取的DNA，同样容易获得来自微卫星基因座的可解释数据。一组六行品种的合作研究也报告了以前未发表的数据和分析。在大麦系Harrington和WM872-3的系谱中，分别检查了11种和5种微卫星随时间的稳定性。一组15条双倍单倍体细胞系中有9个微卫星。在这些测试微卫星线中没有检测到新的微卫星等位基因形成。在这些实验中，在三个品种中对复合二聚体微卫星HVM40和完美的含GA微卫星HVM68检测到了品种内微卫星变异，从而说明了微卫星标记的敏感性，并表明在使用它们之前应先测量品种内微卫星变异。分子标记。设计用于扩增大麦AG / CT的微卫星的九种引物已成功用于扩增六倍体小麦，黑麦和燕麦品种中的序列。在九个引物对中的两个中，在12个小麦品种中检测到多态性，为谷物中的微卫星保守提供了初步证据。

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  • 作者

    Dorocicz, Irene R.;

  • 作者单位

    University of Guelph (Canada).;

  • 授予单位 University of Guelph (Canada).;
  • 学科 Biology Molecular.;Biology Genetics.
  • 学位 Ph.D.
  • 年度 1999
  • 页码 146 p.
  • 总页数 146
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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