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

作　　者：莫天宇 徐善斌 邹德堂[1] 王敬国[1] 刘化龙[1] 孙健[1] 贾琰[1] 赵宏伟[1] 郑洪亮[1] MO Tianyu;XU Shanbin;ZOU Detang;WANG Jingguo;LIU Hualong;SUN Jian;JIA Yan;ZHAO Hongwei;ZHENG Hongliang(Key Laboratory of Germplasm Enhancement, Physiology and Ecology of Food Crops in Cold Region, Ministry of Education, Northeast Agricultural University, Harbin 150030, China)

机构地区：[1]东北农业大学寒地粮食作物种质创新与生理生态教育部重点实验室,黑龙江哈尔滨150030

出　　处：《华北农学报》2021年第1期71-80,共10页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金项目(31601377);黑龙江自然科学基金联合引导项目(LH2019C035);“东农”学者计划青年才俊项目(17QC02)。

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T_(0)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T_(1)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T_(2)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。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 T_(0)via PCR and sequencing method,one of them was double genes mutant plant.In the T_(1)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 T_(2).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.

关 键 词：水稻 盐胁迫 OsEIL1 OsEIL2 CRISPR/Cas9 

分 类 号：Q78[生物学—分子生物学] S511.03[农业科学—作物学]