摘要
为探究月季类钙调神经素B亚基蛋白RcCBL3在维持Na^(+)稳态中的作用及其调控机制,成功克隆了月季RcCBL3基因,并分析其结构特征和表达模式。结果表明:RcCBL3蛋白含有4个EF-Hand结构域,定位于液泡膜系统,且在叶片中高表达。在高浓度Na^(+)处理下,RcCBL3基因表达显著上调,而在高浓度K^(+)处理下,其表达无显著变化。病毒诱导的基因沉默(VIGS)实验表明,RcCBL3基因沉默株系对Na^(+)胁迫的耐受性显著降低,表现为叶片枯萎和相对电导率升高。酵母双杂交和双分子荧光互补实验进一步证实,RcCBL3蛋白能够与RcCIPK1/2/3蛋白相互作用。沉默RcCBL3基因后,RcCIPK1基因表达量上升,而RcCIPK3基因表达量下降,暗示RcCBL3蛋白可能与RcCIPK1/3蛋白共同参与维持细胞Na^(+)稳态。该研究可为揭示月季细胞Na^(+)稳态调控网络提供理论基础,也可为月季耐盐育种工作提供指导意见。
To investigate the role and regulatory mechanism of the Chinese rose calmodulin B‑like protein RcCBL3 in maintaining Na^(+)homeostasis,the Chinese rose RcCBL3 gene was successfully cloned and its structural characteristics and expression patterns were analyzed.The results showed that the RcCBL3 protein contained 4 EF‑Hand domains,located in the vacuolar membrane system,and was highly expressed in leaves.Under high concentration Na^(+)treatment,the expression of RcCBL3 gene was significantly upregulated,while under high concentration K^(+)treatment,its expression showed no significant change.The virus‑induced gene silencing(VIGS)experiment showed that the tolerance of RcCBL3 gene silencing plants to Na^(+)stress significantly reduced,manifested by leaf withering and increased relative conductivity.Further yeast two‑hybrid and bimolecular fluorescence complementation assays confirmed that the RcCBL3 protein could interact with RcCIPK1/2/3 protein,and silencing the RcCBL3 gene resulted in an increase of RcCIPK1 gene expression and a decrease of RcCIPK3 gene expression,suggesting that the RcCBL3 protein might participate in maintaining cellular Na^(+)homeostasis together with RcCIPK1/3 protein.This study could provide a theoretical basis for revealing the Na^(+)homeostasis regulatory network in Chinese rose cells,and also provide guidance for salt tolerant breeding in Chinese roses.
作者
杜俊
王换换
刘蓉
王淞
迟娇娇
奉树成
明凤
DU Jun;WANG Huanhuan;LIU Rong;WANG Song;CHI Jiaojiao;FENG Shucheng;MING Feng(College of Life Sciences,Shanghai Normal University/Development Center of Plant Germplasm Resources,Shanghai Normal University,Shanghai 200234,China;Shanghai Greening Management Guidance Station,Shanghai 200020,China)
出处
《上海农业学报》
2025年第4期1-10,共10页
Acta Agriculturae Shanghai
基金
上海市花卉产业体系建设项目[沪农科产字(2022)第8号]
上海市科技兴农项目(2022-02-08-00-12-F01146)。
