摘要
采用RT-PCR方法从拟南芥叶片中克隆FT基因,经过测序分析、酶切之后,连接到植物表达载体Super 1300+,构建植物重组载体FT-Super 1300+,运用农杆菌介导法将FT基因导入切花菊‘神马’中,鉴定其在转化植株体内的整合和表达。扩增得到的基因片段经测序分析与GenBbank上的FT基因同源性为100%;构建的植物表达载体经过酶切分析证实外源基因已经正确插入;转化后得到了29株抗性植株,PCR和PCR–Southern杂交结果显示,8株抗性植株为阳性,说明外源基因整合到转化植株的基因组中。RT-PCR鉴定结果表明外源基因在转化植株叶片中表达。其中转FT基因的一个株系在组培条件下分化出花芽,表明转基因植株花芽分化不受光周期影响,花期可以提前。
FT gene was cloned by RT-PCR from wild type Arabidopsis.After sequencing,the FT gene with proper digestion sites was digested and inserted into plant expression vector Super 1300＋,and was introduced into chrysanthemum genome by Agrobacterium-mediated transformation.The integration of FT was determined using PCR and PCR–Southern,and gene expression was tested by RT-PCR.After transformation,we obtained 29 resistant plants.PCR and PCR-Southern blot analysis showed that eight of them were positive.RT-PCR detected the expression of the exogenous FT gene in the leaves of transgenic plants.One line of eight transplants appeared flowering bud in vitro.It indicated that transplants flowered early without influence of light.
出处
《园艺学报》
CAS
CSCD
北大核心
2010年第3期441-448,共8页
Acta Horticulturae Sinica
基金
国家‘863’计划项目(2006AA100109)
农业部‘948’项目(2008-G3)
