海藻糖酶基因RNA干扰载体对花烟草的转化

Transformation of Nicotiana alata with RNAi Vector of Trehalase Gene

为了能够使花烟草更好地应用于园林美化,需要进一步提高其抵抗非生物胁迫的能力。海藻糖在植物抗逆性方面具有重要作用。利用RNA干扰技术抑制植物海藻糖酶基因的表达,可以阻断海藻糖降解过程,从而使植物体内海藻糖含量增加,有望以此提高植物抵抗非生物胁迫的能力。本实验利用叶盘侵染法将带有拟南芥海藻糖酶基因干扰载体(iTre-1285)的农杆菌导入花烟草中,经过抗生素筛选初步获得阳性植株后,对阳性植株提取DNA,完成了目的基因的PCR鉴定,初步证明目的基因已转入花烟草植株中。通过本试验,在建立起花烟草基因转化方法的同时,成功地将海藻糖酶干扰基因转入花烟草中。转基因花烟草植株的获得为后期能够进一步筛选出具有抵抗非生物胁迫的转基因植株奠定了基础。

In order to make better use of flower tobacco in landscaping, its ability to resist abiotic stress need to be further enhanced. Trehalose plays an important role in plant resistance. Using RNA interference to inhibit the expression of trehalase gene can block the process of trehalose degradation, so as to increase the trehalose content in plant, which can be expected to improve plant resistance to abiotic stress. In this experiment,agrobacterium carried the RNA interference vector of Arabidopsis trehalase gene（iTre- 1285） had been transformed into flower tobacco（Nicotiana alata） by leaf disc infection method; positive plants were acquired after screening with kanamycin and carbenicillin. DNA was extracted from the positive plants and the target gene appeared in PCR detection. This meant that the target gene had been transformed into flower tobacco plants. So through this study, the genetic transformation methods of flower tobacco had established, while RNAi trehalase gene transformed into flower tobacco successfully. This result laid a foundation for cultivating transgenic flower tobacco against abiotic stress for further.