论文部分内容阅读
【目的】探究Ac SERK1启动子的功能,有助于了解Ac SERK1的表达调控模式。【方法】以‘神湾’菠萝(Ananas comosus L.‘Shenwan’)为材料,将Ac SERK1启动子缺失序列与GUS融合,构建植物表达载体,并导入根瘤农杆菌GV3101中。利用农杆菌真空渗透法侵染烟草叶片,并检测GUS活性;利用浸染法转化菠萝胚性愈伤组织获得转基因植株,分析光照、2,4-D和4℃等处理后的GUS表达量。【结果】构建2个Ac SERK1启动子植物缺失表达载体,分别命名为p(-30/+258 bp)和p(-499/+258 bp)。烟草瞬时表达结果显示p(-499/+258 bp)表现出强的GUS活性,p(-30/+258 bp)表现出微弱的GUS活性。q RT-PCR结果表明,光照处理后,GUS表达量降低。相反的,2,4-D和4℃处理转基因菠萝植株后GUS表达量均显著增加。【结论】Ac SERK1启动子-499/-30 bp区段内含有光、生长素和低温响应元件。
【Objective】 To explore the function of Ac SERK1 promoter and help to understand the expression regulation mode of Ac SERK1. 【Method】 Ananas comosus L.’Shenwan ’was used as a material to fuse Ac SERK1 promoter with GUS. The plant expression vector was constructed and introduced into Agrobacterium tumefaciens GV3101. The Agrobacterium tumefaciens vacuum infiltration method was used to infect tobacco leaves, and the activity of GUS was detected. The transgenic plants were transformed with embryogenic calli by dip dyeing method, and the expression of GUS after 2,4-D and 4 ℃ treatment was analyzed. 【Result】 Two Ac SERK1 promoter-null expression vectors were constructed and named as p (-30 / + 258 bp) and p (-499 / + 258 bp), respectively. Transient expression of tobacco showed p (-499 / + 258 bp) showed strong GUS activity, p (-30 / + 258 bp) showed weak GUS activity. q RT-PCR results show that after light treatment, GUS expression decreased. In contrast, GUS expression was significantly increased after treatment with 2,4-D and 4 ° C transgenic pineapple plants. 【Conclusion】 Ac SERK1 promoter contains light, auxin and low-temperature response elements in -499 / -30 bp segment.