The use of semi-automated fluorescent microsatellite analysis for tomato cultivar identification
The use of semi-automated fluorescent microsatellite analysis for tomato cultivar identification
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DOI：
10.1007/s001220050934
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发表时间：
1998-09-01
影响因子：
5.4
通讯作者：
Vosman, B
中科院分区：
文献类型：
--
作者：
Bredemeijer, GMM;Arens, P;Vosman, B
本研究的目的是评估荧光分析方法对基于 PCR 的番茄微卫星标记（或 STMS）进行基因分型的有用性，并确定这些标记在生成番茄品种独特 DNA 图谱方面的价值。使用荧光染料标记的正向引物并使用 ALF 表达 DNA 测序仪进行分析。一般来说，番茄 STMS 的分析揭示了不同的等位基因峰。在几个番茄 STMS 标记中观察到诸如口吃和差异扩增等 PCR 假象，但在大多数情况下，这些假象并没有严重妨碍等位基因的指定。荧光和银染等位基因图谱的比较揭示了测试品种之间等位基因的相似分布。使用荧光方法对 16 个番茄品种的 20 个选定的 STMS 标记进行 DNA 分型。使用其中 18 个标记检测 PCR 产物的长度多态性，产生 0.06 至 0.74 的基因多样性值。每个微卫星位点的等位基因数量为 2 至 8 个。只需 4 个 STMS 就足以区分所有 16 个品种。表明这些标记特别适合番茄等其他类型标记检测到的变异水平较低的物种。
The objectives of this study were to evaluate the usefulness of a fluorescent-analysis method for genotyping PCR-based tomato microsatellite markers (Or STMSs) and to establish the value of these markers to generate unique DNA profiles of tomato cultivars. The analyses were performed using forward primers labelled with a fluorochrom and using an ALF express DNA sequencer. In general, analysis of the tomato STMSs revealed distinct allelic peaks. PCR artefacts like stuttering and differential amplification were observed for several tomato STMS markers, but in most cases these artefacts did not seriously hamper allele designation. Comparison of fluorescent and silver-stained allelic profiles revealed a similar distribution of alleles among the test cultivars. Sixteen tomato cultivars were DNA-typed for 20 selected STMS markers using the fluorescent approach. Length polymorphism among the PCR products was detected with 18 of these markers, yielding gene diversity values from 0.06 to 0.74. The number of alleles per microsatellite locus ranged from 2 to 8. As few as four STMSs were sufficient to differentiate between all 16 cultivars. indicating that these markers are especially suitable for a species like tomato which has low levels of variation as detected by other types of markers.