利用CRISPR/Cas9技术敲除OsNramp5创制镉低积累水稻新种质
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1.四川省农业科学院生物技术核技术研究所;2.四川省农业科学院水稻高粱研究所

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国家自然科学基金青年项目(32001531);四川省财政“1+3”项目(2021ZYGG);四川省科技计划项目(2021YFYZ0016、2021YFYZ0027)


Generating low Cadmium accumulation new rice germplasms by editing OsNramp5 using CRISPR/Cas9 technology
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Affiliation:

1.Institute of Biotechnology and Nuclear Technology,Sichuan Academy of Agricultural Sciences,Chengdu;2.Rice and Sorghum Research Institute, Sichuan Academy of Agricultural Sciences

Fund Project:

National Natural Science Foundation of China (32001531);Sichuan Provincial Financial “1+3” Project (2021ZYGG);Sichuan Science and Technology Program(2021YFYZ0016、2021YFYZ0027)

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    摘要:

    大米镉超标问题严重威胁人体健康。水稻镉吸收转运基因OsNramp5的功能缺失可有效降低稻米中Cd含量的积累。为了快速创制镉低积累的水稻新种质,本研究采用CRISPR/Cas9基因编辑技术敲除三系杂交稻优质抗病恢复系‘R491’中的镉吸收转运基因OsNramp5,获得了多种不同突变方式的编辑植株,并筛选出单靶点突变的无转基因成份的两种纯合突变株系(KO1和KO2) 。在镉污染土壤中种植并测定野生型和敲除植株糙米的镉含量结果显示,相比于野生型R491,敲除株系KO1和KO2糙米中的镉含量显著下降,均降低了90%左右。农艺性状调查结果发现,相比野生型R491,KO1突变株系的农艺性状没有发生显著差异,但KO2突变株系在株高、结实率和千粒重显著降低。因此,利用CRISPR/Cas9基因编辑技术敲除镉吸收转运基因OsNramp5可快速创制镉低积累的水稻新种质,本研究创制的新种质为加速培育能在Cd污染区种植的安全水稻品种提供了新的遗传资源。

    Abstract:

    The problem of cadmium pollution in rice grain is a serious threat to human health. Disrupting the function of the cadmium transport gene OsNramp5 in rice can effectively reduce the accumulation of Cd in rice. To rapidly create low Cd-accumulating rice germplasm. In this study, we used CRISPR/Cas9 gene editing technology to knock out the cadmium transport gene OsNramp5 under the background of ChuanHui491 (R491), an indica restorer line of three-line hybrid rice with high-quality and disease-resistant. Various edited plants with different mutations were obtained, and two homozygous mutant lines (KO1 and KO2) without transgenic elements with single-target mutations were screened. Compared with wild-type R491, the cadmium content of brown rice in knockout lines KO1 and KO2 was significantly reduced by about 90% when planted in cadmium-polluted soil fields. The results of the agronomic trait investigation showed that there was no significant difference in the KO1 plants compared with wild-type R491, however, the KO2 plants had significant decreases in plant height, seed setting rate, and 1000-grain weight. Therefore, knockout of the cadmium transporter gene OsNramp5 by CRISPR/Cas9 gene editing technology can rapidly create new rice germplasm with low cadmium accumulation, and the new germplasm created in this study provides a new genetic resource for accelerating the breeding of safe rice varieties that can be planted in the cadmium-polluted field.

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  • 收稿日期:2023-11-23
  • 最后修改日期:2024-03-01
  • 录用日期:2024-04-12
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