Knockout of OsHMA3 in an indica rice increases cadmium sensitivity and inhibits plant growth

IF 3.5 3区生物学 Q1 PLANT SCIENCES Plant Growth Regulation Pub Date : 2024-03-16 DOI:10.1007/s10725-024-01137-x

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Abstract

Cadmium (Cd) is a heavy metal that poses harm to both plants and humans. OsHMA3, a member of the heavy metal ATPase (HMA) family, plays a crucial role in sequestering Cd into the vacuoles of roots, thereby limiting its accumulation in rice grains. However, the response of rice plants to Cd under complete loss-of-function of OsHMA3 remains unclear. In this study, we successfully generated OsHMA3 null mutants in an indica variety 93 − 11 using CRISPR/Cas9 technology. A Cd resistance experiment revealed that the Oshma3 mutants exhibited increased sensitivity to Cd compared to the wild-type at a tested concentration of 10 µM CdCl₂. Furthermore, the seedlings of Oshma3 mutant lines displayed inhibited plant growth in the presence of 1 µM Cd, specifically suppressing aboveground growth. As expected, knockout lines of OsHMA3 showed lower Cd accumulation in roots but higher concentrations in shoots compared to wild-type plants, highlighting the role of OsHMA3 in root-to-shoot Cd translocation. We further performed RNA sequencing analysis on wild-type and Oshma3 plants under control and Cd treatment conditions and found that differentially expressed genes were mainly enriched in metal ion binding, integral component of the membranes, and biosynthesis pathways for secondary metabolites triggered by exposure to Cd. When grown in a paddy field, the Oshma3 mutants exhibited shorter plant height, lower seed setting rate, and higher Cd accumulation in grains compared to wild-type plants. Our results indicate that knockout of OsHMA3 in the 93 − 11 variety increases sensitivity to Cd and inhibits plant growth.

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在籼稻中敲除 OsHMA3 会增加对镉的敏感性并抑制植物生长

摘要镉（Cd）是一种对植物和人类都有害的重金属。OsHMA3 是重金属 ATPase（HMA）家族的成员，在将镉封存到根部液泡中从而限制镉在水稻籽粒中的积累方面起着至关重要的作用。然而，在 OsHMA3 功能完全缺失的情况下，水稻植株对镉的反应仍不清楚。在本研究中，我们利用 CRISPR/Cas9 技术成功地在籼稻品种 93 - 11 中产生了 OsHMA3 空突变体。抗镉实验表明，在测试浓度为 10 µM CdCl2 时，与野生型相比，Oshma3 突变体对镉的敏感性增加。此外，在 1 µM Cd 存在下，Oshma3 突变株系的幼苗表现出植物生长受抑制，特别是地上部生长受抑制。正如预期的那样，与野生型植株相比，OsHMA3基因敲除株系在根部的镉积累较低，但在芽中的镉浓度较高，这突出表明了OsHMA3在根到芽的镉转运中的作用。我们进一步对对照和镉处理条件下的野生型植株和Oshma3植株进行了RNA测序分析，发现差异表达的基因主要富集在金属离子结合、膜的整体成分和次生代谢物的生物合成途径中。与野生型植株相比，Oshma3突变体在水稻田中生长时表现出较矮的株高、较低的结实率和较高的籽粒镉积累。我们的研究结果表明，在 93 - 11 品种中敲除 OsHMA3 会增加对镉的敏感性并抑制植物生长。

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来源期刊

Plant Growth Regulation 生物-植物科学

CiteScore

6.90

自引率

9.50%

发文量

139

审稿时长

4.5 months

期刊介绍： Plant Growth Regulation is an international journal publishing original articles on all aspects of plant growth and development. We welcome manuscripts reporting question-based research using hormonal, physiological, environmental, genetical, biophysical, developmental or molecular approaches to the study of plant growth regulation. Emphasis is placed on papers presenting the results of original research. Occasional reviews on important topics will also be welcome. All contributions must be in English.