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miRNA广泛参与植物生长发育的调控,但与植物活体再生相关的miRNA研究还未见报道。本研究以不依赖外源激素发生在番茄活体植株伤口上的再生体系为实验材料,构建了愈伤组织分化过程中的小RNA文库。通过高通量测序,在对照组与活体再生组分别鉴定了82个与75个novel miRNA,其中36个差异性表达。差异表达的miRNA共预测到2 159个靶基因,通过靶基因注释,发现两个miRNA,Novel-56和Novel-128,调控与植物顶端分生组织生长相关的NAM基因。设计茎环引物对两个miRNA进行了鉴定。本研究首次鉴定了植物活体再生中的novel miRNA表达谱,揭示了活体再生中特异表达的novel miRNA,有助于更好的理解番茄活体再生机制。
MiRNAs are widely involved in the regulation of plant growth and development, but miRNAs related to plant regeneration have not yet been reported. In this study, a regeneration system that does not depend on exogenous hormones on the wounds of tomato living plants was used as experimental material to construct a small RNA library during callus differentiation. By high-throughput sequencing, 82 and 75 novel miRNAs were identified in the control and live regeneration groups, respectively, of which 36 were differentially expressed. Differentially expressed miRNAs predicted a total of 2 159 target genes. Two target miRNAs, Novel-56 and Novel-128, were found to target NAM genes involved in apical meristem growth by annotating target genes. Two stem-loop primers were designed to identify two miRNAs. This study, for the first time, identified novel miRNA expression profiles in plant regeneration and revealed novel miRNAs that are specifically expressed during in vivo regeneration, helping to better understand the mechanism of live regeneration in tomato.