Simultaneous genetic transformation and genome editing of mixed lines in soybean (Glycine max) and maize (Zea mays).

IF 4.6 4区 农林科学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY aBIOTECH Pub Date : 2024-06-18 eCollection Date: 2024-06-01 DOI:10.1007/s42994-024-00173-5
Michelle Valentine, David Butruille, Frederic Achard, Steven Beach, Brent Brower-Toland, Edward Cargill, Megan Hassebrock, Jennifer Rinehart, Thomas Ream, Yurong Chen
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Abstract

Robust genome editing technologies are becoming part of the crop breeding toolbox. Currently, genome editing is usually conducted either at a single locus, or multiple loci, in a variety at one time. Massively parallel genomics platforms, multifaceted genome editing capabilities, and flexible transformation systems enable targeted variation at nearly any locus, across the spectrum of genotypes within a species. We demonstrate here the simultaneous transformation and editing of many genotypes, by targeting mixed seed embryo explants with genome editing machinery, followed by re-identification through genotyping after plant regeneration. Transformation and Editing of Mixed Lines (TREDMIL) produced transformed individuals representing 101 of 104 (97%) mixed elite genotypes in soybean; and 22 of 40 (55%) and 9 of 36 (25%) mixed maize female and male elite inbred genotypes, respectively. Characterization of edited genotypes for the regenerated individuals identified over 800 distinct edits at the Determinate1 (Dt1) locus in samples from 101 soybean genotypes and 95 distinct Brown midrib3 (Bm3) edits in samples from 17 maize genotypes. These results illustrate how TREDMIL can help accelerate the development and deployment of customized crop varieties for future precision breeding.

Supplementary information: The online version contains supplementary material available at 10.1007/s42994-024-00173-5.

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大豆(Glycine max)和玉米(Zea mays)混合品系的同步遗传转化和基因组编辑。
强大的基因组编辑技术正在成为作物育种工具箱的一部分。目前,基因组编辑通常是在一个品种的单个基因座或多个基因座上同时进行。大规模并行基因组学平台、多方面的基因组编辑能力和灵活的转化系统使我们能够在一个物种的整个基因型谱中对几乎任何位点进行有针对性的变异。我们在这里展示了利用基因组编辑机器对混合种子胚胎外植体进行定向改造和编辑,然后在植物再生后通过基因分型进行重新鉴定,从而实现多种基因型的同时转化和编辑。混合品系的转化和编辑(TREDMIL)产生的转化个体代表了 104 个大豆混合精英基因型中的 101 个(97%);以及 40 个玉米雌性和雄性混合精英近交系基因型中的 22 个(55%)和 36 个中的 9 个(25%)。对再生个体的编辑基因型进行鉴定,在 101 个大豆基因型的样本中发现了 800 多个不同的确定性1(Dt1)基因座编辑,在 17 个玉米基因型的样本中发现了 95 个不同的棕色中肋3(Bm3)编辑。这些结果说明了 TREDMIL 如何帮助加速开发和部署定制的作物品种,以实现未来的精准育种:在线版本包含补充材料,可查阅 10.1007/s42994-024-00173-5。
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来源期刊
CiteScore
7.70
自引率
2.80%
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0
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