Generation of low-cadmium rice germplasms via knockout of OsLCD using CRISPR/Cas9
Generation of low-cadmium rice germplasms via knockout of OsLCD using CRISPR/Cas9
复制标题
使用 CRISPR/Cas9 敲除 OsLCD 生成低镉水稻种质
DOI：
10.1016/j.jes.2022.05.047
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发表时间：
2022-06-14
影响因子：
6.9
通讯作者：
Yi, Jicai
中科院分区：
文献类型：
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作者：
Chen, Huamei;Ye, Rong;Yi, Jicai
OSLCD基因与镉（CD）在大米中的积累有关，可能是CRISPR/CAS9编辑的有用靶标。但是，OSLCD基因编辑对CD积累，植物生长和产量特征的影响尚不清楚。在这里，我们使用CRISPR/CAS9生成来自Indica和Japonica水稻品种的OSLCD单突变体。我们还生成了Indica背景中的OSNRAMP5单突变体和OSLCD OSNRAMP5双突变体。当在cd污染的稻田中生长时，所有OSLCD单个突变体的cd累积少于野生类型（WTS）。与此相比，与此相比，与WTS相比，在CD污染的水培培养物中生长的OSLCD单突变体在芽中积累的CD明显较小。积累的减少可能是由于CD应力下CD易位的减少而导致的。与CD存在的WT相比，与WT相比，所有OSLCD单一突变幼苗的氧化损伤也会减少，并且植物的生长增加。 OSLCD单突变体和WT之间相似，植物生长和大多数产量性状以及大米幼苗，糙米和稻草的必要元素浓度也相似。在存在CD的情况下，与WTS以及OsnRamp单个突变体相比，糙米中的CD浓度和OSLCD OSNRAMP5双突变体的芽降低。我们的结果表明，OSLCD敲除可以单独减少CD的积累，也可以与各种水稻基因型中的其他基因敲除突变相结合。与OSNRAMP5突变不同，OSLCD敲除并不能减少基本元素含量。因此，OSLCD敲除可以用于产生低CD水稻种系。 （c）2022年中国科学院生态环境科学研究中心。由Elsevier B.V.出版
The OsLCD gene, which has been implicated in cadmium (Cd) accumulation in rice, might be a useful target for CRISPR/Cas9 editing. However, the effects of OsLCD gene editing on Cd accumulation, plant growth, and yield traits remain unknown. Here, we used CRISPR/Cas9 to generate oslcd single mutants from indica and japonica rice cultivars. We also generated osnramp5 single mutants and oslcd osnramp5 double mutants in the indica background. When grown in Cd-contaminated paddy soils, all oslcd single mutants accumulated less Cd than the wild types (WTs). Consistent with this, oslcd single mutants grown in Cd-contaminated hydroponic culture accumulated significantly less Cd in the shoots as compared to WTs. This decrease in accumulation probably resulted from the reduction of Cd translocation under Cd stress. Oxidative damage also decreased, and plant growth increased in all oslcd single mutant seedlings as compared to WTs in the presence of Cd. Plant growth and most yield traits, as well essential element concentrations in rice seedling shoots, brown rice, and rice straw, were similar between oslcd single mutants and WTs. In the presence of Cd, Cd concentrations in the brown rice and shoots of oslcd osnramp5 double mutants were sig-nificantly decreased compared with WTs as well as osnramp single mutants. Our results suggested that OsLCD knockout may reduce Cd accumulation alone or in combination with other knockout mutations in a variety of rice genotypes; unlike OsNramp5 mutations, OsLCD knockout did not reduce essential element contents. Therefore, OsLCD knockout might be used to generate low-Cd rice germplasms. (c) 2022 The Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences. Published by Elsevier B.V.