甘蔗ScWRKY4基因的克隆与表达特性分析

王玲,刘峰,戴明剑,孙婷婷,苏炜华,王春风,张旭,毛花英,苏亚春,阙友雄

Cloning and Expression Characteristic Analysis of ScWRKY4 Gene in Sugarcane

Ling WANG,Feng LIU,Ming-Jian DAI,Ting-Ting SUN,Wei-Hua SU,Chun-Feng WANG,Xu ZHANG,Hua-Ying MAO,Ya-Chun SU,You-Xiong QUE

图6 甘蔗ScWRKY4亚细胞定位分析 (A) 35S::ScWRKY4::GFP亚细胞定位重组载体的酶切验证结果。1: 15000+2000 bp DNA marker; 2: 35S::GFP/Xba I; 3: ScWRKY4 ORF PCR产物; 4: 35S::ScWRKY4::GFP/Xba I; 5: 35S::ScWRKY4::GFP/Xba I+BamH I; 6: 100 bp ladder DNA marker。(B) 农杆菌介导转化的ScWRKY4及空载体注射本氏烟叶片48 h的亚细胞定位结果。本氏烟叶片表皮细胞被用于明场、绿色荧光、蓝色荧光、明场和绿色及蓝色荧光叠加后的图像分析; 白色箭头1、2和3分别表示质膜、细胞核和细胞质, 比例尺=25 μm; 35S::GFP: 携带空载pCAMBIA 1300-GFP的农杆菌菌株; 35S::ScWRKY4::GFP: 携带重组载体pCAMBIA 1300-ScWRKY4-GFP的农杆菌菌株; DAPI: 4’,6-二脒基-2-苯基吲哚。

Fig. 6 Subcellular localization assay of sugarcane ScWRKY4 (A) The enzyme digestion to identify the insert-integrated subcellular localization expression vector 35S::ScWRKY4::GFP. 1: 15000+2000 bp DNA marker; 2: 35S::GFP/Xba I; 3: ScWRKY4 ORF PCR product; 4: 35S::ScWRKY4::GFP/Xba I; 5: 35S::ScWRKY4::GFP/Xba I+BamH I; 6: 100 bp ladder DNA marker. (B) Subcellular localizations of the Agrobacterium mediated transformation of ScWRKY4 and empty vector in Nicotiana benthamiana leaves 48 h after infiltration. The epidermal cells of N. benthamiana were used for taking images of visible light, green fluorescence, blue fluorescence, merged visible light and green fluorescence and blue fluorescence; white arrows 1, 2, and 3 indicate plasma membrane, nucleus and cytoplasm, respectively; scale bar = 25 μm; 35S::GFP: the Agrobacterium tumefaciens strain carrying the empty vector pCAMBIA 1300-GFP; 35S::ScWRKY4::GFP: the A. tumefaciens strain carrying the recombinant vector pCAMBIA 1300-ScWRKY4-GFP; DAPI: 4’,6-diamidino-2-phenylindole.