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利用韩国甜瓜地方品种的下一代测序技术开发 SSR 标记。

Development of SSR markers by next-generation sequencing of Korean landraces of chamoe (Cucumis melo var. makuwa).

机构信息

Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahangno, Daejeon, 305-806, Republic of Korea.

出版信息

Mol Biol Rep. 2013 Dec;40(12):6855-62. doi: 10.1007/s11033-013-2803-0. Epub 2013 Oct 5.

Abstract

The oriental melon (Cucumis melo var. makuwa), called 'chamoe' in Korean, is a popular fruit crop cultivated mainly in Asia and a high-market value crop in Korea. To provide molecular breeding resources for chamoe, we developed and characterized genomic SSR markers from the preliminary Illumina read assemblies of Gotgam chamoe (one of the major landraces; KM) and SW3 (the breeding parent). Mononucleotide motifs were the most abundant type of markers, followed by di-, tri-, tetra-, and pentanucleotide motifs. The most abundant dinucleotide was AT, followed by AG and AC, and AAT was the most abundant trinucleotide motif in both assemblies. Following our SSR-marker development strategy, we designed a total of 370 primer sets. Of these, 236 primer sets were tested, exhibiting 93 % polymorphism between KM and SW3. Those polymorphic SSRs were successfully amplified in the netted and Kirkagac melons, which respectively exhibited 81 and 76 % polymorphism relative to KM, and 32 and 38 % polymorphism relative to SW3. Seven selected SSR markers with a total of 17 alleles (2-3 alleles per locus) were used to distinguish between KM, SW3, and four chamoe cultivars. Our results represent the first attempt to provide genomic resources for Korean landraces for the purposes of chamoe breeding, as well as to discover a set of SSR markers capable of discriminating chamoe varieties from Korea and the rest of Asia, which possess little genetic diversity. This study establishes a highly efficient strategy for developing SSR markers from preliminary Illumina assemblies of AT-rich genomes.

摘要

东方甜瓜(Cucumis melo var. makuwa),在韩语中称为“chamoe”,是一种主要在亚洲种植的受欢迎的水果作物,也是韩国高市场价值的作物。为了为 chamoe 提供分子育种资源,我们从 Gotgam chamoe(主要地方品种之一;KM)和 SW3(育种亲本)的初步 Illumina 读取组装中开发和表征了基因组 SSR 标记。单核苷酸基序是最丰富的标记类型,其次是二核苷酸、三核苷酸、四核苷酸和五核苷酸基序。最丰富的二核苷酸是 AT,其次是 AG 和 AC,AAT 是两个组装体中最丰富的三核苷酸基序。按照我们的 SSR 标记开发策略,我们总共设计了 370 对引物。其中,236 对引物进行了测试,在 KM 和 SW3 之间表现出 93%的多态性。这些多态性 SSR 在 netted 和 Kirkagac 甜瓜中成功扩增,相对于 KM 分别表现出 81%和 76%的多态性,相对于 SW3 分别表现出 32%和 38%的多态性。从七个选定的 SSR 标记中总共发现了 17 个等位基因(每个位点 2-3 个等位基因),用于区分 KM、SW3 和四个 chamoe 品种。我们的结果代表了首次尝试为韩国地方品种提供 chamoe 育种的基因组资源,并发现了一组 SSR 标记,能够区分韩国和亚洲其他地区的 chamoe 品种,这些品种的遗传多样性很小。这项研究建立了一种从富含 AT 的基因组的初步 Illumina 组装中开发 SSR 标记的高效策略。

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