甘蔗脂氧合酶基因ScLOX1的克隆与表达分析

孙婷婷,王文举,娄文月,刘峰,张旭,王玲,陈玉凤,阙友雄,许莉萍,李大妹,苏亚春

Cloning and expression analysis of sugarcane lipoxygenase gene ScLOX1

Ting-Ting SUN,Wen-Ju WANG,Wen-Yue LOU,Feng LIU,Xu ZHANG,Ling WANG,Yu-Feng CHEN,You-Xiong QUE,Li-Ping XU,Da-Mei LI,Ya-Chun SU

图8 ScLOX1基因在本氏烟叶片中瞬时表达 A: RT-PCR检测ScLOX1基因在接种携带35S::00和35S::ScLOX1载体的农杆菌菌株GV3101的本氏烟叶片1 d的表达情况。B: 对接种携带35S::00和35S::ScLOX1载体的农杆菌2 d的本氏烟叶片进行DAB染色。(1)和(2)分别代表索尼相机和显微镜拍摄的图像。C: 烟草免疫相关标记基因在接种携带35S::00和35S::ScLOX1载体的农杆菌1 d的本氏烟叶片中的相对表达水平。D和F: 瞬时过表达ScLOX1基因1 d后接种烟草青枯菌和茄病镰刀菌蓝色变种1 d和7 d时的本氏烟叶片发病情况和DAB染色结果。(1)和(2)分别代表索尼相机和显微镜拍摄的图像。E和G: 烟草免疫相关标记基因在瞬时过表达ScLOX1基因的本氏烟叶片接种青枯菌和茄病镰刀菌蓝色变种1 d和7 d时的表达情况分析。烟草免疫相关标记基因包括HR标记基因NtHSR201、NtHSR203和NtHSR515, SA途径相关基因NtPR-1a/c, JA途径相关基因NtPR2和NtPR3, ET合成依赖基因NtEFE26和NtAccdeaminase。内参基因为NtEF-1α基因。柱上不同的字母代表在P < 0.05时显著性的差异。所有的点均为平均值±标准误 (n = 3)。a和b分别表示载体35S::00和35S::ScLOX1。

Fig. 8 Transient overexpression of ScLOX1 in Nicotiana benthamiana leaves A: RT-PCR analysis of ScLOX1 gene in the N. benthamiana leaves after infiltration by Agrobacterium strain GV3101 carrying vector 35S::00 or 35S::ScLOX1 for 1 d. B: DAB staining with N. benthamiana leaves 2 d after 35S::ScLOX1-containing Agrobacterium strain infiltration. (1) and (2) represent images taken by SONY camera and microscope, respectively. C: Relative expression level of the tobacco immunity-associated marker genes in 35S::ScLOX1-transiently expressing leaves after infiltration for 1 d. D and F: The results of disease symptoms and DAB staining of N. benthamiana leaves after inoculation with 35S::ScLOX1 1 d and then infection with Ralstonia solanacearum and Fusarium solani var. coeruleum for 1 d and 7 d, respectively. (1) and (2) represent images taken by SONY camera and microscope, respectively. E and G: Analysis of the immunity related marker genes in the N. benthamiana leaves by R. solanacearum and F. solani var. coeruleum for 1 d and 7 d post inoculation. The tobacco immunity-associated marker genes including the HR marker genes NtHSR201, NtHSR203 and NtHSR515, the SA pathway related genes NtPR-1a/c, the JA pathway-associated genes NtPR2 and NtPR3, and the ET synthesis depended genes NtEFE26 and NtAccdeaminase. Using NtEF-1α as an internal reference gene. Bars superscripted by different lowercase letters are significantly different at P < 0.05. Data points are means ± SE (n = 3). The vectors of 35S::00 and 35S::ScLOX1 are indicated by a and b, respectively.