为水稻(Oryza sativa)多样化育种应用设计的数千种特异性 KASP 标记。

IF 2.1 3区 生物学 Q3 GENETICS & HEREDITY G3: Genes|Genomes|Genetics Pub Date : 2024-11-01 DOI:10.1093/g3journal/jkae251
Katherine Steele, Mark Quinton-Tulloch, Darshna Vyas, John Witcombe
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引用次数: 0

摘要

本研究旨在通过在引物设计中加入模糊(退化)碱基,扩大 KASP 在不同基因型水稻中的适用性,并验证其中 4000 个引物在基因分型中的应用。利用生物信息学管道将来自 89 个国家的 129 个水稻基因组与籼稻参考基因组 R498 进行了比较,并针对 R498 和其他基因组之间更常见的变异生成了 160 万个 KASP 设计。在这些设计中,98 238 个是预测的功能标记。在由 178 个不同水稻品种组成的小组中,对 1024 个育种者选择的位点各进行了多达 5 个 KASP 的检测,产生了 3366 个有效的 KASP。84% 的成功率在 KASP 的正常范围内,这表明模糊碱基不会影响其有效性。3366 个性状特异性标记小组被用于多样性小组的群体结构分析,并将其分为四个预期群体。将用于设计的 13 个基因组序列中的目标变异与多样性面板中相同的 13 个品种的其他加入物的相应 KASP 基因型进行了比较。在 12 个品种中,79% 的标记具有一致性。有 10 个品种的一致性很高(>88%),但从一个陆地品种中选出的一个品种的一致性仅为 46.5%。育种人员现在可以在三个备选参考基因组(包括 Niponbare MSU7)中搜索经过验证的 KASP 和超过 100 万个迄今为止未经测试的设计,搜索与以前发表的 SSR 标记接近的设计,并检索 129 个水稻基因组中的目标变异及其基因组位置和 +/-25 bp 侧翼序列。
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Thousands of trait-specific KASP markers designed for diverse breeding applications in rice (Oryza sativa).

This study aimed to broaden applicability of KASP for Oryza sativa across diverse genotypes through incorporation of ambiguous (degenerate) bases into their primer designs and to validate 4000 of them for genotyping applications. A bioinformatics pipeline was used to compare 129 rice genomes from 89 countries with the indica reference genome R498 and generate ∼1.6 million KASP designs for the more common variants between R498 and the other genomes. Of the designs, 98,238 were for predicted functional markers. Up to five KASP each for 1024 breeder-selected loci were assayed in a panel of 178 diverse rice varieties, generating 3366 validated KASP. The 84% success rate was within the normal range for KASP demonstrating that the ambiguous bases do not compromise efficacy. The 3366-trait-specific marker panel was applied for population structure analysis in the diversity panel and resolved them into four expected groups. Target variations in thirteen of the genome sequences used for designs were compared with the corresponding KASP genotypes of other accessions of the same thirteen varieties in the diversity panel. There was agreement across 12 varieties for 79% of markers. Ten varieties had high agreement (>88%) but a variety selected from a landrace had only 46.5% agreement. Breeders can now search for the validated KASP and >1 million so-far untested designs across three alternative reference genomes (including Niponbare MSU7), search for designs proximal to previously published SSR markers and retrieve the target variations in 129 rice genomes plus their genomic locations with +/-25 bp flanking sequences.

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来源期刊
G3: Genes|Genomes|Genetics
G3: Genes|Genomes|Genetics GENETICS & HEREDITY-
CiteScore
5.10
自引率
3.80%
发文量
305
审稿时长
3-8 weeks
期刊介绍: G3: Genes, Genomes, Genetics provides a forum for the publication of high‐quality foundational research, particularly research that generates useful genetic and genomic information such as genome maps, single gene studies, genome‐wide association and QTL studies, as well as genome reports, mutant screens, and advances in methods and technology. The Editorial Board of G3 believes that rapid dissemination of these data is the necessary foundation for analysis that leads to mechanistic insights. G3, published by the Genetics Society of America, meets the critical and growing need of the genetics community for rapid review and publication of important results in all areas of genetics. G3 offers the opportunity to publish the puzzling finding or to present unpublished results that may not have been submitted for review and publication due to a perceived lack of a potential high-impact finding. G3 has earned the DOAJ Seal, which is a mark of certification for open access journals, awarded by DOAJ to journals that achieve a high level of openness, adhere to Best Practice and high publishing standards.
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