title: The cbfs triple mutants reveal the essential functions of CBFs in cold acclimation and allow the definition of CBF regulons in Arabidopsis.
abstract: In Arabidopsis, the C-repeat binding factors (CBFs) have been extensively studied as key transcription factors in the cold stress response. However, their exact functions in the cold response remains unclear due to the lack of a null cbf triple mutant. In this study, we used CRISPR/Cas9 technology to mutate CBF1 or CBF1/CBF2 in a cbf3 T-DNA insertion mutant to generate cbf1,3 double and cbf1 cbf2 cbf3 (cbfs) triple mutants. The response of the cbfs triple mutants to chilling stress is impaired. Furthermore, no significant difference in freezing tolerance was observed between the wild-type and the cbf1,3 and cbfs mutants without cold acclimation. However, the cbfs mutants were extremely sensitive to freezing stress after cold acclimation, and freezing sensitivity ranking was cbfs > cbf1,3 > cbf3. RNA-Seq analysis showed that 134 genes were CBF regulated, of which 112 were regulated positively and 22 negatively by CBFs. Our study reveals the essential functions of CBFs in chilling stress response and cold acclimation, as well as defines a set of genes as CBF regulon. It also provides materials for the genetic dissection of components in CBF-dependent cold signaling.
authors.affiliation: State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.
authors.forename: Yuxin
authors.initials: Y
authors.lastname: Jia
authors.affiliation: State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.
authors.forename: Yanglin
authors.initials: Y
authors.lastname: Ding
authors.affiliation: State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.
authors.forename: Yiting
authors.initials: Y
authors.lastname: Shi
authors.affiliation: State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.
authors.forename: Zhizhong
authors.initials: Z
authors.lastname: Gong
authors.affiliation: State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, 100193, China. yangshuhua@cau.edu.cn.
authors.forename: Shuhua
authors.initials: S
authors.lastname: Yang
mesh_terms: D000064:Acclimatization / Q000235:genetics*; D017360:Arabidopsis / Q000235:genetics* / Q000502:physiology*; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003080:Cold Temperature; D005615:Freezing; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D009154:Mutation / Q000235:genetics*; D018085:Regulon / Q000235:genetics*; D017423:Sequence Analysis, RNA; D013312:Stress, Physiological / Q000235:genetics; D059467:Transcriptome / Q000235:genetics
chemical_list: D029681:Arabidopsis Proteins
keywords: Arabidopsis; C-repeat binding factor (CBF) transcription factors; CBF regulons; chilling response; cold acclimation