水稻OsCDF1基因突变效应及其基因组变异分析  

Mutation effects of OsCDF1 gene and its genomic variations in rice

作　　者：胡艳娟 薛丹 耿嫡 朱末 王天穹 王晓雪[1] HU Yan-Juan;XUE Dan;GENG Di;ZHU Mo;WANG Tian-Qiong;WANG Xiao-Xue(Rice Research Institute,Shenyang Agricultural University,Shenyang 110866,Liaoning,China)

机构地区：[1]沈阳农业大学水稻研究所,辽宁沈阳110866

出　　处：《作物学报》2023年第9期2362-2372,共11页Acta Agronomica Sinica

基　　金：国家重点研发计划项目(2017YFD0300107);国家自然科学基金项目(32070642)资助。

摘　　要：开花期(又叫抽穗期)影响水稻的产量、品质和地区适应性。在拟南芥中, Cycling DOF Factor 1 (CDF1)蛋白质是CONSTANS(CO)的转录抑制因子,负向调节拟南芥的开花期。但是,水稻中的同源蛋白OsCDF1的功能尚不完全清楚。为了揭示OsCDF1在水稻中的生物学功能及其对开花期的影响,本研究利用CRISPR/Cas9基因编辑技术设计定向敲除水稻OsCDF1基因的2个靶位点,构建OsCDF1基因的敲除载体;通过农杆菌介导的方法分别转化北方粳稻品种沈农9816,创制OsCDF1基因的突变体;分析沈农9816和oscdf1突变体在田间种植下的开花期和产量性状的差异。主要研究结果为:创制了在第1个外显子的第16bp处缺失5个碱基和在第2个外显子的338bp处单碱基A插入的oscdf1纯合突变。序列比对分析表明,这2种类型的突变均造成移码和蛋白翻译提前终止。在自然长日照条件下, oscdf1突变体的开花期比野生型沈农9816晚4 d以上,其产量高于野生型。对OsCDF1单倍型和单倍型网络分析发现, OsCDF1在不同品种中进化出高度多样性。本研究成功利用CRISPR/Cas9基因编辑技术敲除了水稻OsCDF1基因,为进一步研究OsCDF1基因功能提供了理论参考,也为水稻遗传改良提供了潜在的基因和种质资源。Flowering time(heading date)affects yield,quality,and regional adaptability of rice.The Cycling DOF Factor 1(CDF1)protein is a transcriptional repressor of CONSTANS(CO)and negatively regulates flowering time in Arabidopsis.However,the biological functions of OsCDF1 in rice is not quite clear.To explore the biological functions of OsCDF1 and its effects on flowering time control in rice,we constructed two binary vectors carrying guide RNAs targeting OsCDF1 gene via CRISPR/Cas9 system.The resultant plasmids were transferred into SN9816 which was the variety widely cultivated in northern China by using an Agrobacterium-mediated transformation,and the mutations of OsCDF1 was firstly generated in SN9816.The flowering time and yield related traits of SN9816 and oscdf1 mutants were investigated in the paddy field.The main results were as follows:Two homozygous oscdf1 lines were identified,including a five bp deletion at 16th bp of the first exon and a single base pair A insertion at 338th bp of the second exon.Sequence alignment analysis revealed that the two types of mutations resulted in frame-shift and premature translation termination.Mutations of OsCDF1 delayed flowering time,but increased yield under natural long day conditions in rice.Analysis of OsCDF1 genetic variations and haplotype networks revealed that the rice accessions had evolved high genomic diversity in OsCDF1 locus.The knockout mutants of OsCDF1 created by CRISPR/Cas9 provided the theoretical basis to further study the role of OsCDF1 gene in rice and the potential gene and germplasm resources for genetic improvement in rice.

关 键 词：水稻 CRISPR/Cas9 OsCDF1 开花期 产量相关性状 单倍型 

分 类 号：S511[农业科学—作物学]