利用CRISPR/Cas9技术敲除OsNramp5创制镉低积累水稻新种质
Generating Low Cadmium Accumulation New Rice Germplasms by Editing OsNramp5 using CRISPR/Cas9 Technology
胡彬华 1蒲志刚 1何志渊 1王平 1白玉路 1李赓觅 2张涛 2蒋开锋 2杨莉2
作者信息
- 1. 四川省农业科学院生物技术核技术研究所,成都 610066
- 2. 四川省农业科学院水稻高粱研究所(四川省农业科学院德阳分院),德阳 618000
- 折叠
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镉在稻米中的积累.为了快速创制镉低积累的水稻新种质,本研究利用CRISPR/Cas9基因编辑技术敲除三系杂交稻优质抗病恢复系川恢491(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,均降低约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显著差异,但KO2突变株系的株高、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可在镉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 excessive cadmium in rice grains seriously threatens human health.Disrupting the function of the cadmium transport gene OsNramp5 in rice can effectively reduce cadmium accumulation in rice.To rapidly create new rice germplasm with low cadmium accumulation,this study used CRISPR/Cas9 gene editing technology to knock out the cadmium transport gene OsNramp5 in the high-quality disease-resistant restorer line Chuanhui491(R491)of three-line hybrid rice.Various edited plants with different mutations were obtained,and two homozygous mutant lines(KO1 and KO2)with single-target mutations without transgenic elements were selected.Compared with wild-type R491,the cadmium content of brown rice in knockout lines KO1 and KO2 were significantly reduced by about 90％when planted in cadmium-polluted soil fields.Agronomic trait investigation revealed no significant difference between the KO1 plants and wild-type R491,whereas the plant height,seed setting rate,and 1000-grain weight of KO2 mutant lines were significantly reduced.Therefore,knocking out the cadmium transporter gene OsNramp5 by CRISPR/Cas9 gene editing technology can quickly create new rice germplasm with low cadmium accumulation.The new germplasm created in this study provides novel genetic resources for accelerating the breeding of safe rice varieties that can be planted in the cadmium-polluted field.
关键词
水稻/镉/OsNramp5/CRISPR/Cas9/育种Key words
rice/cadmium/OsNramp5/CRISPR/Cas9/breeding引用本文复制引用
基金项目
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)
四川省科技计划项目(2021YFYZ0027)
出版年
2024
国家科技期刊平台
NSTL
万方数据