通过CRISPR介导的BnaNRAMP1同源拷贝编辑创制低镉积累且产量不受显著影响的油菜种质

doi:10.1016/j.jia.2024.05.016

Journal of Integrative Agriculture ›› 2025, Vol. 24 ›› Issue (5): 1704-1717.DOI: 10.1016/j.jia.2024.05.016

通过CRISPR介导的BnaNRAMP1同源拷贝编辑创制低镉积累且产量不受显著影响的油菜种质

收稿日期:2024-02-03 修回日期:2024-05-13 接受日期:2024-04-02 出版日期:2025-05-20 发布日期:2025-04-14

CRISPR-mediated editing of BnaNRAMP1 homologous copies creates a low Cd-accumulation oilseed rape germplasm with unaffected yield

Ying Zhang^1*, Rui Wang^1*#, Tianshun Luo¹, Jingyan Fu¹, Meng Yin¹, Maolin Wang¹, Yun Zhao^{1, 2#}

¹Key Laboratory for Bio-Resources and Eco-Environment, College of Life Sciences, Sichuan University, Chengdu 610065, China
²Science and Technology Innovation Center of Sichuan Modern Seed Industry Group, Chengdu 611100, China

Received:2024-02-03 Revised:2024-05-13 Accepted:2024-04-02 Online:2025-05-20 Published:2025-04-14
About author:Ying Zhang, E-mail: zhangying2021@stu.scu.edu.cn; #Correspondence Rui Wang, E-mail: wangray1987@scu.edu.cn; Yun Zhao, E-mail: zhaoyun@scu.edu.cn * These authors contributed equally to this study.
Supported by:
This research was supported by the National Key Research and Development Program of China (2022YFD1601502 and 2023YFD1200202), the National Natural Science Foundation of China (32272167 and 32341028), the Sichuan Science and Technology Program, China (2022ZDZX0015), and the Fundamental Research Funds for the Central Universities, China (SCU2019D013).

摘要/Abstract

摘要：

甘蓝型油菜是种植于全世界的最重要的油料作物之一，土壤镉（Cd）污染显著影响其产量和种子品质。甘蓝型油菜具有强镉吸附能力，因此创制低镉积累的种质资源是缓解这一挑战的重要策略。本研究中，我们分别靶向BnaNRAMP1的三个不同外显子区，对其在油菜进行基因编辑，通过基因编辑植株的系列表型鉴定筛选出镉积累显著降低且产量不受显著影响的油菜种质资源K140-22。此外，通过对K140-22根和地上部的抗氧化防御系统的相关酶活进行测定，发现其抗氧化活性增强，这有助于阐明植物耐受重金属胁迫的分子机制。值得注意的是，K140-22的产量性状和种子品质相较于野生型均没有显著差异，显示出其在镉污染土壤提高油菜经济价值及低镉积累品种选育中的应用潜力。

Abstract:

Brassica napus, one of the most important oil crops cultivated globally, is severely impacted by prolonged soil contamination with cadmium (Cd), resulting in decreased yields and poor seed quality. This crop exhibits a high adsorption capacity for Cd, making creating seed resources with low Cd accumulation an essential strategy to alleviate this challenge. To address this issue, we genetically edited BnaNRAMP1 in B. napus by targeting three different exon regions, resulting in new germplasm resources with significant differences in Cd accumulation capacity and unaffected yield. Among these, the mutant K140-22, specifically targeting the 7th exon, is distinguished by its substantially reduced Cd accumulation. Further, enzyme assays of the antioxidant defense system in both roots and shoots of K140-22 revealed its enhanced antioxidant activity, which contributes to elucidating the molecular mechanisms of plant tolerance to heavy metal stress. Remarkably, this mutant also maintained equivalent agronomic traits and seed quality, which highlights its potential as a germplasm resource for rapeseed breeding for low Cd accumulation and elevating rapeseed economic value in Cd-contaminated soil.

Key words: Brassica napus , Cd accumulation , BnaNRAMP1-edited lines , gene editing

Ying Zhang, Rui Wang, Tianshun Luo, Jingyan Fu, Meng Yin, Maolin Wang, Yun Zhao. 通过CRISPR介导的BnaNRAMP1同源拷贝编辑创制低镉积累且产量不受显著影响的油菜种质[J]. Journal of Integrative Agriculture, 2025, 24(5): 1704-1717.

Ying Zhang, Rui Wang, Tianshun Luo, Jingyan Fu, Meng Yin, Maolin Wang, Yun Zhao. CRISPR-mediated editing of BnaNRAMP1 homologous copies creates a low Cd-accumulation oilseed rape germplasm with unaffected yield[J]. Journal of Integrative Agriculture, 2025, 24(5): 1704-1717.

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通过CRISPR介导的BnaNRAMP1同源拷贝编辑创制低镉积累且产量不受显著影响的油菜种质

CRISPR-mediated editing of BnaNRAMP1 homologous copies creates a low Cd-accumulation oilseed rape germplasm with unaffected yield

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