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采用RACE-PCR法,从‘小樱桃’文心兰中克隆到一个全长989bp的铁氧还蛋白基因cDNA序列,命名为OnFd(登录号KX461907)。OnFd基因开放阅读框长为465bp,预测可编码154个氨基酸;gDNA和cDNA序列比对结果显示OnFd基因不含内含子。生物信息学分析表明,OnFd具有1个典型的[2Fe-2S]结构域;同源分析显示,OnFd与玉米Fd3的相似度最高(64.29%)。蛋白亚细胞定位结果显示OnFd定位于叶绿体。实时荧光定量PCR检测发现:OnFd基因在花中表达量最高,其次是叶与根,在假鳞茎中表达量最低;接种病原菌研究显示,文心兰感染软腐病后,各个部位的OnFd基因表达量均呈上升趋势,尤其是接种部位假鳞茎在接种病原菌1d后表达量便表现出极显著上调,并且在5个感病阶段的表达量是健康植株的2.83~3.98倍。研究表明,OnFd基因可能在文心兰响应抗软腐病过程中具有重要作用。
A total length of 989bp of the ferredoxin gene cDNA sequence was cloned from Oncorn avenaceae using RACE-PCR and named OnFd (accession number KX461907). OnFd gene open reading frame length of 465bp, predicted to encode 154 amino acids; gDNA and cDNA sequence alignment showed that OnFd gene does not contain introns. Bioinformatics analysis showed that OnFd has a typical [2Fe-2S] domain. Homology analysis showed that OnFd had the highest similarity (64.29%) with maize Fd3. Protein subcellular localization results show that OnFd is located in the chloroplast. The results of real-time fluorescent quantitative PCR showed that OnFd gene was the highest in flowers, followed by leaves and roots, and the lowest in pseudobulb. Inoculation with pathogen showed that OnFd gene expression in various parts of Inula sinica infected with soft rot Especially the inoculated part of pseudobulb showed a significant up-regulation after 1d of inoculation with pathogen, and the expression level in the five susceptible stages was 2.83 ~ 3.98 times of healthy plants. Studies have shown that OnFd gene may play an important role in the response of Oncidium against soft rot.