通过 CRISPR/Cas9 基因病毒复制子系统生成的水稻收获前萌芽抗性突变体的表型特征。

IF 2.9 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
BMB Reports Pub Date : 2024-02-01
Jong Hee Kim, Jihyeon Yu, Jin Young Kim, Yong Jin Park, Sangsu Bae, Kwon Kyoo Kang, Yu Jin Jung
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引用次数: 0

摘要

收获前萌发是一种关键现象,涉及种子在相对潮湿和休眠降低的条件下收获前在母株中的萌发。在本文中,我们利用 CRISPR/Cas9 和 geminiviral 复制子系统,在 OsERF1 基因中产生了一个区域 SNP(C/T)和一个插入 6 bp(GGT/GGTGGCGGC)的 HDR 突变株,以抗性收获前萌发(PHS)。在转化的胼胝体中,HDR 发生率为 2.6%。从 12 个 HDR 诱导的胼胝体中收获了 T1 种子,并将其命名为 ERF1-hdr 品系。对ERF1-hdr品系的分子稳定性、关键农艺特性、生理特性和目标基因的生化特性进行了为期三年的研究。qRT-PCR 分析表明,ABA 信号的增强导致了更强的 PHS 抗性表型。这些结果表明,利用适用于不同水稻目标和其他作物的单一模块化配置,可以通过 SNP/InDel 替换实现高效 HDR。这项工作证明了在一代人的时间内用精英等位基因替换所有基因的潜力,并极大地扩展了我们改良重要农业性状的能力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Phenotypic characterization of pre-harvest sprouting resistance mutants generated by the CRISPR/Cas9-geminiviral replicon system in rice.

Pre-harvest sprouting is a critical phenomenon involving germination of seeds in the mother plant before harvest under relative humid conditions and reduced dormancy. In this paper, we generated HDR mutant lines with one region SNP (C/T) and an insertion of 6 bp (GGT/GGTGGCGGC) in OsERF1 genes for pre-harvest sprouting (PHS) resistance using CRISPR/Cas9 and a geminiviral replicon system. The incidence of HDR was 2.6% in transformed calli. T1 seeds were harvested from 12 HDR-induced calli and named ERF1-hdr line. Molecular stability, key agronomic properties, physiological properties, and biochemical properties of target genes in the ERF1-hdr line were investigated for three years. The ERF1-hdr line showed significantly enhanced seed dormancy and pre-harvest sprouting resistance. qRT-PCR analysis suggested that enhanced ABA signaling resulted in a stronger phenotype of PHS resistance. These results indicate that efficient HDR can be achieved through SNP/InDel replacement using a single and modular configuration applicable to different rice targets and other crops. This work demonstrates the potential to replace all genes with elite alleles within one generation and greatly expands our ability to improve agriculturally important traits. [BMB Reports 2024; 57(2): 79-85].

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来源期刊
BMB Reports
BMB Reports 生物-生化与分子生物学
CiteScore
5.10
自引率
7.90%
发文量
141
审稿时长
1 months
期刊介绍: The BMB Reports (BMB Rep, established in 1968) is published at the end of every month by Korean Society for Biochemistry and Molecular Biology. Copyright is reserved by the Society. The journal publishes short articles and mini reviews. We expect that the BMB Reports will deliver the new scientific findings and knowledge to our readers in fast and timely manner.
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