利用CRISPR/Cas9技术敲除<i>OsEIL1</i>和<i>OsEIL2</i>基因改良水稻耐盐性

doi:10.7668/hbnxb.20190798

摘要/Abstract

摘要： 为了培育耐盐水稻品种，利用CRISPR/Cas9基因编辑技术，在水稻品种东农427的耐盐负调控基因OsEIL1和OsEIL2设计1个敲除靶点对其进行同时敲除。通过PCR和测序的方法，在T₀共检测出了5个突变单株，其中有1株为双基因同时突变的植株。在该植株的T₁株系中，获得了4株双基因纯合突变且不含T-DNA元件插入的植株。在T₂进行苗期耐盐性鉴定，成熟期考察农艺性状指标。结果表明，在苗期200 mmol/L NaCl溶液处理5 d后，突变体植株的鲜质量和干质量显著高于野生型植株，地上部及地下部钠离子含量均显著低于野生型植株，钾离子含量与野生型植株无明显差异。恢复7 d后，突变体植株幼苗存活率（75.0%）显著高于野生型植株（8.3%）。在成熟期，突变体的主要农艺性状未发生明显变化。综上，利用CRISPR/Cas9技术对水稻品种东农427进行了耐盐改良，为耐盐水稻品种的培育提供了理论和材料基础。

关键词: 水稻, 盐胁迫, OsEIL1, OsEIL2, CRISPR/Cas9

Abstract: In order to breeding salt tolerant rice varieties, a target was designed to simultaneously knockout OsEIL1 and OsEIL2 genes in rice variety Dongnong 427 by using CRISPR/Cas9 technology. Five mutant plants were obtained in T₀ via PCR and sequencing method, one of them was double genes mutant plant. In the T₁ mutant lines of this plant, four double gene homozygous mutant plants without T-DNA element were obtained. Salt tolerance experiment in seedling stage and agronomic trait analysis at maturity were carried out in T₂. The results indicated that after 200 mmol/L NaCl solution treatment for 5 days, the fresh weight and the dry weight of mutant plants were significantly higher than wild type plants, the Na⁺ content in shoot and root were significantly lower than wild type plants, however, the K⁺ content was no difference with wild type plants. After recovered for 7 days, the seedling survival rate of mutant plants(75.0%) was significantly higher than wild type plants(8.3%). The main agronomic traits didn't change significantly at maturity. In summary, this study improved the salt tolerance of rice variety Dongnong 427 by using CRISPR/Cas9 technology, and provided a theoretical and material basis for the cultivation of salt tolerant rice variety.

Key words: Rice, Salt tolerance, OsEIL1, OsEIL2, CRISPR/Cas9

中图分类号:

Q78
S511.03

莫天宇, 徐善斌, 邹德堂, 王敬国, 刘化龙, 孙健, 贾琰, 赵宏伟, 郑洪亮. 利用CRISPR/Cas9技术敲除OsEIL1和OsEIL2基因改良水稻耐盐性[J]. 华北农学报, 2021, 36(1): 71-80. doi: 10.7668/hbnxb.20190798.

MO Tianyu, XU Shanbin, ZOU Detang, WANG Jingguo, LIU Hualong, SUN Jian, JIA Yan, ZHAO Hongwei, ZHENG Hongliang. Enhancing Salt Tolerance of Rice by Knocking out OsEIL1 and OsEIL2 Via CRISPR/Cas9 System[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(1): 71-80. doi: 10.7668/hbnxb.20190798.

http://www.hbnxb.net/CN/Y2021/V36/I1/71

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利用CRISPR/Cas9技术敲除OsEIL1和OsEIL2基因改良水稻耐盐性

Enhancing Salt Tolerance of Rice by Knocking out OsEIL1 and OsEIL2 Via CRISPR/Cas9 System

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利用CRISPR/Cas9技术敲除OsEIL1和OsEIL2基因改良水稻耐盐性

Enhancing Salt Tolerance of Rice by Knocking out OsEIL1 and OsEIL2 Via CRISPR/Cas9 System

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