Automated sizing of fluorescent-labeled Simple Sequence Repeat (SSR) markers to assay genetic variation in soybean
Automated sizing of fluorescent-labeled Simple Sequence Repeat (SSR) markers to assay genetic variation in soybean
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DOI：
10.1007/s001220050618
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发表时间：
1997-10-01
影响因子：
5.4
通讯作者：
Cregan, PB
中科院分区：
文献类型：
--
作者：
Diwan, N;Cregan, PB
简单重复序列;微卫星（SSR）等位基因大小测定为基因型鉴定、系谱分析和估计生物体之间的遗传距离提供了有用的工具。大豆[Glycine max（L.）梅尔。]为了植物品种保护（PVP）的目的，通过标准色素和形态性状鉴定栽培品种。然而，许多商业大豆来自有限数量的优良品系，并且通常无法根据这些性状进行区分。基于SSR标记的系统将提供品种的独特DNA图谱。等位基因的荧光标记结合每个凝胶泳道中的内部大小标准的自动大小测定被用作标准[P-32]标记的替代，以评估大豆的遗传变异性。20个SSR位点的等位基因频率在35个大豆基因型，占95%以上的等位基因在北美大豆品种系谱分析的基础上确定。在20个SSR位点上，平均每个位点有10.1个等位基因（范围：5-17），平均基因多样性为0.80（范围：0.50 - 0.87）。这20个基因座成功地区分了形态和色素性状相同的现代大豆品种，以及7个大豆基因型报告使用17个RFLP探针无法区分。以7个大豆品种为材料，研究了SSR标记在大豆遗传中的世代稳定性。在7个家系中，有6个家系的后代中有一个基因座的等位基因在双亲中均不存在。这些新的等位基因很可能是突变的结果。大豆SSR等位基因突变率与人类相似。为了避免与突变相关的困难，DNA指纹数据应该从一个栽培品种的30-50株植物中确定。
Simple Sequence Repeat; (SSR) allele sizing provides a useful tool for genotype identification, pedigree analysis, and for estimating genetic distance between organisms. Soybean [Glycine max (L.) Merr.] cultivars are identified for Plant Variety Protection (PVP) purposes by standard pigmentation and morphological traits. However, many commercial soybeans arise from a limited number of elite lines and are often indistinguishable based on these traits. A system based on SSR markers would provide unique DNA profiles of cultivars. Fluorescent labeling of alleles combined with automated sizing with internal size standards in each gel lane was used as an alternative to standard [P-32] labeling to assess genetic variability in soybean. Allelic frequencies at 20 SSR loci were determined in 35 soybean genotypes that account for greater than 95% of the alleles in North American soybean cultivars based upon pedigree analysis. An average of 10.1 alleles per locus (range: 5-17), with a mean gene diversity of 0.80 (range: 0.50 to 0.87) were observed at the 20 SSR loci. The 20 loci successfully distinguished modern soybean cultivars that are identical for morphological and pigmentation traits, as well as 7 soybean genotypes reported to be indistinguishable using 17 RFLP probes. Pedigrees of 7 cultivars were studied to estimate stability of SSRs in soybean across generations. Of the 7 pedigrees 6 had one locus in the progeny with an allele(s) that was not present in either parent. These new alleles are most likely the result of mutation. The mutation rate of SSR alleles in soybean was similar to that reported in humans. To avoid difficulty associated with mutation, DNA fingerprint data should be determined from the bulk of 30-50 plants of a cultivar.