马铃薯StNCED1基因过表达载体的构建及其遗传转化

Construction of Overexpression Vector of StNCED1 Gene and Its Genetic Transformation of Potato

马铃薯块茎的休眠和发芽对于块茎生产和加工极为重要,延长块茎休眠期有助于降低块茎水分和养分的消耗,从而提高其应用价值。本研究用PCR方法从马铃薯中克隆了9-顺式-环氧类胡萝卜素双氧合酶(NCED)编码基因St NCED1,并构建了马铃薯块茎特异表达启动子CIPP驱动的St NCED1基因的植物过表达载体p BIC-St NCED1,然后通过根癌农杆菌介导法转化马铃薯栽培品种"陇薯3号"获得转化植株,经卡那霉素抗性筛选得到5株抗性苗,PCR检测证明St NCED1基因已整合到马铃薯的基因组中。qRT-PCR检测表明St NCED1基因在转基因植株试管薯中的表达量分别比未转基因的对照增加2.10~8.09倍。该研究有助于培育块茎休眠期延长的马铃薯新种质。

Abstrac t Dormancy and sprouting of potato tubers are very important for tuber production and processing.Prolonging dormancy period of tuber is helpful for reducing consumption of water and nutrients so as to improve tuber application value. In this research, the St NCED1 gene coded 9-cis-epoxycarotenoid dioxygenase（NCED）was cloned through PCR method. The plant overexpression vector p BIC-St NCED1 was constructed which was driven by tuber-specific expression promoter CIPP. The transformed plants of potato cultivar ＂Longshu 3＂were obtained by Agrobacterium-mediated method. 5 kanamycin-resistant plants were obtained by kanamycin screening and the PCR assay showed that St NCED1 gene had been integrated into the genome of potato. qRT-PCR analysis indicated that the relative expression amount of St NCED1 gene in microtubers of the transgenic plants increased from 2.10 to 8.09-fold compared with the non-transgenic plants. This research could be useful for breeding new potato germplasm with prolonged period of tuber dormancy.