玉米ZmRBOHD1和ZmCDPK32的蛋白相互作用研究  

Identification of ZmRBOHD1 interacting with ZmCDPK32 in maize

作　　者：李洁[1] 初智霖 李小蒙 周晓今[2] LI Jie;CHU Zhilin;LI Xiaomeng;ZHOU Xiaojin(College of Life Sciences,Beijing Normal University,100875,Beijing,China;Biotechnology Research Institute,Chinese Academy of Agricultural Sciences,100081,Beijing,China)

机构地区：[1]北京师范大学生命科学学院,北京100875 [2]中国农业科学院生物技术研究所,北京100081

出　　处：《北京师范大学学报（自然科学版）》2023年第4期629-636,共8页Journal of Beijing Normal University(Natural Science)

基　　金：国家自然科学基金资助项目(31771707)。

摘　　要：为探索玉米呼吸爆发氧化酶(RBOH)与钙依赖型蛋白激酶(CDPK)协同调控花粉管极性生长的机制,通过蛋白保守结构域分析获得14个ZmRBOH家族基因,且利用转录组数据分析了ZmRBOH的时空表达模式.结果显示,ZmRBOHD1和ZmRBOHD2在玉米成熟花粉中特异表达.选择花粉特异表达的ZmRBOHD1和功能已知的花粉管极性生长调节因子ZmCDPK32开展蛋白相互作用研究,结论有如下2点:1)通过瞬时表达ZmRBOHD1-GFP融合蛋白证明ZmRBOHD1定位于细胞膜;2)利用酵母双杂交和双分子荧光互补技术证实ZmRBOHD1可以和ZmCDPK32相互作用,并且生物信息学分析发现ZmRBOHD1具有潜在的CDPK磷酸化靶点.结果表明,ZmCDPK32可能通过与ZmRBOHD1的相互作用协同调控玉米花粉管发育,这为深入探究花粉管极性生长的分子机制奠定了基础.Both respiratory burst oxidase homologue(RBOH)and calcium dependent protein kinase(CDPK)regulate pollen tube polar growth,but whether these two proteins interact remains unclear.To explore the mechanism of ZmRBOH and ZmCDPK synergistic regulation of pollen tube polar growth,14 ZmRBOH family genes we identified from the maize genome,temporal and spatial expression patterns of ZmRBOH genes were examined from transcriptome data.Expression analyses found that ZmRBOHD1 and ZmRBOHD2 were specifically expressed in mature maize pollen.The interaction between pollen specifically expressed ZmRBOHD1 and a known pollen tube polar growth regulator,ZmCDPK32,was investigated.Transient expression of ZmRBOHD1-GFP fusion protein demonstrated that ZmRBOHD1 was localized to cell membranes.Yeast two-hybrid and bimolecular fluorescence complementation techniques confirmed that ZmRBOHD1 interacted with ZmCDPK32.Bioinformatics analysis revealed potential CDPK phosphorylation motifs in ZmRBOHD1.The above data together suggest that ZmCDPK32 synergically regulates pollen tube development in maize by interacting with ZmRBOHD1.Further study of the molecular mechanism of pollen tube polar growth may be needed.

关 键 词：玉米 呼吸爆发氧化酶 钙依赖型蛋白激酶 酵母双杂交 双分子荧光互补 

分 类 号：Q945.45[生物学—植物学]