红豆越桔VviDREB1基因转化烟草的研究

Study on Transgenic Tobacco with Vaccinium vitis-idaea VviDREB1 Gene

通过构建pVB4215植物双元表达载体,采用农杆菌介导法转化烟草,研究VviDREB1在植物体中的异源表达特性.结果显示,实验获得了25个Hyg抗性株系,经过PCR、RT-PCR和GUS组织化学染色检测及Hyg基因的PCR复检等多点验证,证实表达载体边界内序列完整地整合到2个烟草株系的基因组中.转基因烟草株系在4℃低温处理20h后,恢复生长5h,叶片光系统PSⅡ抗寒性分析结果表明,转基因植株的叶片快速叶绿素荧光曲线OJIP各点数值高于对照植株,VviDREB1基因能够显著提高烟草的荧光产量,最大光化学效率Fv/Fm和以吸收光能为基础的性能指数PIABS较对照高,说明VviDREB1对保护植物组织细胞内光合系统PSⅡ有明显的作用,转VviDREB1基因烟草对低温有一定的忍耐能力.

VviDREB1 （Vaccinium vitis-idaea Dehydration Responsive Element Binding 1） is a transcription factor in Vaccinium vitis-idaea Linn.related to abiotic stresses.To research expression characterization of VviDREB1,an overexpression vector of pVB4215 was constructed and transferred it into tobacco,25 Hyg positive tobacco plants were obtained.Analysis of PCR and RT-PCR of VviDREB1,GUS straining and Hyg PCR showed that there was the whole sequence of expression vector in the genomes of the two transgenic tobaccos.This provided the reliable transgenic experience materials for the gene functional analysis.After 4℃ for 20 h and then 25℃ for 5 h,the cold resistance analysis result of the photosystem Ⅱ of transgenic tobacco leaves showed that the fast induction curves of chlorophyll fluorescence of transgenic tobacco was higher than that of the control,the VviDREB1 could improve the fluorescence yield of the transgenic tobacco,the maximal photochemical efficiency Fv/Fm and the performance based on light absorption PIABS were improved comparing with that of the control,which play an obvious role for the protection of photosynthetic organization,and improved its cold resistance.