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开发和利用基于通用 PCR 和高通量 KASP 标记的黑麦(Secale cereale L.)染色体臂的方法。

Scale development and utilization of universal PCR-based and high-throughput KASP markers specific for chromosome arms of rye (Secale cereale L.).

机构信息

Center for Agricultural Resources Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Shijiazhuang, 050021, Hebei, China.

University of Chinese Academy of Sciences, Beijing, 100049, China.

出版信息

BMC Genomics. 2020 Mar 4;21(1):206. doi: 10.1186/s12864-020-6624-y.

Abstract

BACKGROUND

Rye (Secale cereale L., 2n = 2x = 14, RR), a relative of common wheat, is a large gene resource pool for wheat improvement. Accurate and convenient identification of the rye chromatin in wheat background will facilitate the transfer and utilization of elite genes derived from rye in wheat breeding.

RESULTS

In the present study, five rye cultivars including Imperial, German White, Jingzhouheimai, Baili and Guyuan were sequenced by specific-locus amplified fragment sequencing (SLAF-seq) to develop large-scale rye-specific markers. Based on SLAF-seq and bioinformatics analyses, a total of 404 universal PCR-based and a whole set of Kompetitive allele-specific PCR (KASP) markers specific for the 14 individual rye chromosome arms were developed and validated. Additionally, two KASP markers specific for 1RS and 2RL were successfully applied in the detection of 1RS translocations in a natural population and 2RL chromosome arms in wheat-rye derived progenies that conferred adult resistance to powdery mildew.

CONCLUSION

The 404 PCR-based markers and 14 KASP markers specific for the 14 individual rye chromosome arms developed in this study can enrich the marker densities for gene mapping and accelerate the utilization of rye-derived genes in wheat improvement. Especially, the KASP markers achieved high-throughput and accurate detection of rye chromatin in wheat background, thus can be efficiently used in marker-assisted selection (MAS). Besides, the strategy of rye-specific PCR-based markers converting into KASP markers was high-efficient and low-cost, which will facilitate the tracing of alien genes, and can also be referred for other wheat relatives.

摘要

背景

黑麦(Secale cereale L.,2n=2x=14,RR)是普通小麦的近亲,是小麦改良的大基因资源库。准确、方便地鉴定小麦背景中的黑麦染色质将有助于在小麦育种中转移和利用来自黑麦的优良基因。

结果

本研究利用特定基因座扩增片段测序(SLAF-seq)对 5 个黑麦品种(Imperial、German White、Jingzhouheimai、Baili 和 Guyuan)进行测序,开发了大规模的黑麦特异性标记。基于 SLAF-seq 和生物信息学分析,共开发和验证了 404 个通用基于 PCR 的标记和一整套 Kompetitive allele-specific PCR(KASP)标记,用于 14 条黑麦单条染色体臂。此外,两个用于 1RS 和 2RL 的 KASP 标记成功应用于自然群体中 1RS 易位和小麦-黑麦衍生后代中 2RL 染色体臂的检测,这些后代赋予了对白粉病的成株抗性。

结论

本研究开发的 404 个基于 PCR 的标记和 14 个针对 14 条黑麦单条染色体臂的 KASP 标记可以丰富基因定位的标记密度,加速黑麦基因在小麦改良中的利用。特别是,KASP 标记实现了对小麦背景中黑麦染色质的高通量和精确检测,因此可有效地用于标记辅助选择(MAS)。此外,黑麦特异性基于 PCR 的标记转化为 KASP 标记的策略高效且低成本,这将有助于追踪外源基因,也可作为其他小麦近缘种的参考。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2014/7057559/74e8e5801741/12864_2020_6624_Fig1_HTML.jpg

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