为探究多花水仙ACS基因的序列特征及功能,以云香'水仙盛花期花瓣为试验材料,根据云香'水仙花朵转录组数据信息,通过RT-PCR方法克隆出1个AC; S基因,命名为NtACS1(GenBank; KX082936);NtACS1开放阅读框(ORF)长度为552bp,编码183个氨基酸。编码蛋白质分子量约为20.6KDa,理论等电点为6.3; 0,不稳定系数为65.49,属于不稳定的疏水性蛋白。通过qRT-PCR对云香'水仙不同时期花瓣和副冠中的NtACS1基因进行了表达分析,得到与云; 香'水仙花朵转录组数据中相同的结果:NtACS1基因在云香'水仙花瓣和副冠中的表达都是随着花衰老过程呈现逐渐下降的趋势,且NtACS1基因在花瓣; 和副冠中的表达峰值都在花苞期,表明; NtACS1基因编码的蛋白是在乙烯生物合成途径的系统1发挥催化作用的ACC合成酶。成功构建了NtACS1基因的正义植物表达载体,并通过农杆菌介导; 法获得8株转基因烟草,PCR和RT-PCR检测显示其中有6株为阳性植株,初步证实NtACS1基因已导入烟草基因组中且在烟草中已表达。该研究结果为; 进一步分析NtACS1基因的功能和后续转化水仙延长其花期研究奠定了基础。Aimed to study the characteristics and functions of ACSgene,in the; present study,we cloned a 1-aminocyclopropane-1-carboxylate synthetase; gene named NtACS1(GeneBank KX082936)based on the RNA-Seq database from; the flower of Narcissus tazetta var.Yunxiang'using RT-PCR method.The; length of the open reading frame(ORF)of ACSis 552bp,encoding 183amino; acids coupled with a molecular weight of 20.6kDa and theoretical; isoelectric point of 6.30.qRT-PCR analysis showed that the relative; expression level of NtACS1both in petals and coronas are decreased; gradually along with the aging of flower.Moreover,the expression data of; NtACS1gene were consistent with those obtained by RNA-Seq, implied that; the NtACS1protein as an ACC synthetase might play a role in the; catalytic system 1 of ethylene biosynthesis.Furthermore,sense plant; expression vectors of NtACS1were successfully constructed with; agrobacterium mediated transformation,and 6positive transgenic tobacco; plants were ultimately obtained. Our current study will lay an; experimental foundation for the future application of the genetic; transformation to prolong florescence ofYunxiang'.福建省种业创新与产业化工程项
