Development of Microsatellite Markers in Bunching Onion (Allium fistulosum L.)
Development of Microsatellite Markers in Bunching Onion (Allium fistulosum L.)
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DOI：
10.1270/jsbbs.54.361
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发表时间：
2004-12
期刊：
影响因子：
2.4
通讯作者：
Yeon-Sang Song;Keita Suwabe;T. Wako;T. Ohara;T. Nunome;A. Kojima
中科院分区：
文献类型：
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作者：
Yeon-Sang Song;Keita Suwabe;T. Wako;T. Ohara;T. Nunome;A. Kojima
由于洋葱中可用的遗传标记很少，因此需要许多 DNA 标记来构建该作物育种的主要基础。我们在这里报告了洋葱成束中微卫星标记的发展。从束状洋葱品种的基因组 DNA 构建了大小分级的基因组文库。 ‘Kujo Futo’并用 (GA)15 和 (GT)15 寡核苷酸探针的混合物进行筛选。我们从大约 180,000 个克隆中分离出 94 个阳性克隆并对其进行了测序。总共 52 个克隆被鉴定为具有微卫星重复。其中 49 种具有 GT 基序，而只有一种具有 GA 基序，已知在大多数植物物种中，GA 基序比 GT 更常见。这是葱属植物中 GT 丰度高于 GA 的第一份报告。为含微卫星的克隆设计了50个特异性PCR引物对，其中33个引物对扩增了9个束洋葱品种的多态性位点，总共检测到115个等位基因。这些结果表明，具有二核苷酸基序的微卫星，尤其是具有 GT 的微卫星，是洋葱束中信息丰富的遗传标记的有希望的来源。
Since few genetic markers are available in bunching onion, many DNA markers are needed for the construction of a primary basis for the breeding of this crop. We report here the development of microsatellite markers in bunching onion. A size-fractionated genomic library was constructed from genomic DNA of bunching onion cv. ‘Kujo Futo’ and screened with a mixture of (GA)15 and (GT)15 oligonucleotide probes. From approximately 180,000 clones we isolated and sequenced 94 positive clones. A total of 52 clones were identified as having microsatellite repeats. Of them 49 had a GT motif, while only one had a GA motif, which is known to be more frequent than GT in most plant species. This is the first report of the abundance of GT over GA in Allium spp. Of 50 specific PCR primer pairs designed for the microsatellite-containing clones, 33 primer pairs amplified polymorphic loci in nine cultivars of bunching onion, 115 alleles detected in total. These results indicate that microsatellites with dinucleotide motifs, especially with GT, are promising sources of highly informative genetic markers in bunching onion.