蜡梅SAMT基因遗传转化及其功能分析

Genetic transformation and functional analysis of the SAMT gene in Chimonanthus praecox

为研究蜡梅SAMT基因的调控功能,通过构建pBI121-CpSAMT植物双元表达载体,利用农杆菌介导法将其转入矮牵牛中,经PCR和RT-PCR对转化植株进行检测,同时采用顶空固相微萃取以及气相色谱-质谱技术(HS-SPME-GC-MS),对转基因矮牵牛鲜花进行花香成分分析。结果显示:9株转化苗均能扩增出目的条带,RT-PCR检测结果进一步证明,阳性植株均发生了正确转录;转基因矮牵牛植株和未转基因对照植株在植株大小、叶片形态、花色、花瓣大小以及花期等方面均未发现有明显差异。鲜花芳香成分分析表明,转CpSAMT基因矮牵牛中苯甲醛和苯乙醇含量明显升高,并产生了香茅醇、乙酸香茅酯和乙酸苯乙酯等成分,但水杨酸甲酯和苯甲酸甲酯含量没有显著改变。

In order to investigate the biological function of theChimonanthus praecox SAMTgene, here we reported thatthe plant binary expression vector pBI121-CpSAMTwas constructed and then theCpSAMTgene was introduced into pe-tunia throughAgrobacterium tumefaciensmediated transformation. PCR and RT-PCR was used to test the transgenicplants, and HS-SPME-GC-MS was employed to analyze the components of flower flavor. Results showed that nine inde-pendent transgenic lines obtained were positive. Further analysis showed that transgenic plants were indistinguishablefrom untransformed controls in terms of plant size, leaf morphology, flower color, petal size and flowering time. The a-nalysis of HS-SPME-GC-MS showed that the content of benzaldehyde, phenylethanol, citronellol, citronellyl acetateand phenethyl acetate in flowers of transgenicCpSAMTplants was increased. However, the contents of methyl salicylateand methyl benzoate had not been significantly changed comparing with non-transgenic controls.