Development and validation of KASP assays for genes underpinning key economic traits in bread wheat
Development and validation of KASP assays for genes underpinning key economic traits in bread wheat
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DOI：
10.1007/s00122-016-2743-x
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发表时间：
2016-10-01
影响因子：
5.4
通讯作者：
He, Zhonghu
中科院分区：
文献类型：
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作者：
Rasheed, Awais;Wen, Weie;He, Zhonghu
我们开发并验证了一个强大的标记工具包，用于高通量和经济高效地筛选小麦中的大量功能基因。功能标记（FM）是作物育种计划中最有价值的标记，FM 的高通量基因分型可以为在育种品种时有效实践标记辅助选择提供绝佳的机会。在这里，我们开发并验证了竞争性等位基因特异性 PCR (KASP) 检测方法，用于检测支撑面包小麦经济重要性状的基因，包括适应性、谷物产量、质量以及生物和非生物胁迫抗性。总共 70 种 KASP 检测方法（无论是本研究中开发的还是从公共数据库获得的）都经过了应用可靠性的验证。 KASP 测定的验证通过以下方式进行：(a) 将测定与可用的基于凝胶的 PCR 标记对 23 个不同的小麦种质进行比较，(b) 使用由来自中国和其他 13 个国家的 300 个不同品种组成的小组的表型验证衍生的等位基因信息，以及 (c) 在可能的情况下对四个分离群体中的测定进行额外测试。报告的所有 KASP 检测均与品种组和双亲群体中的相关表型显着相关，从而揭示了其在小麦育种计划中的潜在应用。结果显示，KASP 检测的速度比基于凝胶的 PCR 标记物优越 45 倍。 KASP 最近作为单重高通量基因分型技术出现；这是第一份关于对主要作物中大量功能基因进行高通量筛选的报告。此类测定可以极大地加速杂交亲本和先进品系的表征，以进行标记辅助选择，并可以补充不灵活的高密度 SNP 阵列。我们的结果提供了一个强大而可靠的分子标记工具包，有助于最大限度地提高小麦育种计划的遗传收益。
We developed and validated a robust marker toolkit for high-throughput and cost-effective screening of a large number of functional genes in wheat.Functional markers (FMs) are the most valuable markers for crop breeding programs, and high-throughput genotyping for FMs could provide an excellent opportunity to effectively practice marker-assisted selection while breeding cultivars. Here we developed and validated kompetitive allele-specific PCR (KASP) assays for genes that underpin economically important traits in bread wheat including adaptability, grain yield, quality, and biotic and abiotic stress resistances. In total, 70 KASP assays either developed in this study or obtained from public databases were validated for reliability in application. The validation of KASP assays were conducted by (a) comparing the assays with available gel-based PCR markers on 23 diverse wheat accessions, (b) validation of the derived allelic information using phenotypes of a panel comprised of 300 diverse cultivars from China and 13 other countries, and (c) additional testing, where possible, of the assays in four segregating populations. All KASP assays being reported were significantly associated with the relevant phenotypes in the cultivars panel and bi-parental populations, thus revealing potential application in wheat breeding programs. The results revealed 45 times superiority of the KASP assays in speed than gel-based PCR markers. KASP has recently emerged as single-plex high-throughput genotyping technology; this is the first report on high-throughput screening of a large number of functional genes in a major crop. Such assays could greatly accelerate the characterization of crossing parents and advanced lines for marker-assisted selection and can complement the inflexible, high-density SNP arrays. Our results offer a robust and reliable molecular marker toolkit that can contribute towards maximizing genetic gains in wheat breeding programs.