Zhang Di, Ren Li, Yue Jian-Hua, Shi Yu-Bo, Zhuo Li-Huan, Wang Ling, Shen Xiao-Hui
Key Laboratory of Urban Agriculture (South) Ministry of Agriculture, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China.
Department of Ornamental Plants and Horticulture, College of Landscape Architecture, Northeast Forestry University, Harbin 150040, China.
Gene. 2015 Jul 10;565(2):252-67. doi: 10.1016/j.gene.2015.04.013. Epub 2015 Apr 10.
Agapanthus praecox is a monocotyledonous ornamental bulb plant. Generally, the scape (inflorescence stem) length can develop more than 1m, however application 400 mg·L(-1) paclobutrazol can shorten the length beyond 70%. To get a deeper insight into its dwarfism mechanism, de novo RNA-Seq technology has been employed, for the first time, to describe the scape transcriptome of A. praecox. We got 71,258 assembled unigenes, and 45,597 unigenes obtained protein functional annotation. Take the above sequencing results as a reference gene set, using RNA-seq (quantification) technology analyzed gene expression profiles between the control and paclobutrazol-treated samples, and screened 2838 differentially expressed genes. GO, KEGG and MapMan pathway analyses indicated that these differentially expressed genes were significantly enriched in response to stimulus, hormonal signaling, carbohydrate metabolism, cell wall, cell size, and cell cycle related biological process. To validate the expression profiles obtained by RNA-Seq, real-time qPCR was performed on 24 genes selected from key significantly enriched pathways. Comprehensive analysis suggested that paclobutrazol blocks GA signal that can effectively inhibit scape elongation; the GA signal interact with other hormonal signals including auxin, ethylene, brassinosteroid and cytokinins, and trigger downstream signaling cascades leading to metabolism, cell wall biosynthesis, cell division and the cycle decreased obviously, and finally induced dwarfism trait. Furthermore, AP2/EREBP, bHLH, C2H2, ARR, WRKY and ARF family's transcription factors were involved in the regulation of scape development in A. praecox. This transcriptome dataset will serve as an important public information platform to accelerate research on the gene expression and functional genomics of Agapanthus.
百子莲是一种单子叶观赏球根植物。一般来说,花葶(花序茎)长度可超过1米,然而施用400毫克·升(-1)多效唑可使长度缩短70%以上。为了更深入了解其矮化机制,首次采用了从头RNA测序技术来描述百子莲的花葶转录组。我们获得了71258个组装的单基因,其中45597个单基因获得了蛋白质功能注释。以上述测序结果作为参考基因集,利用RNA测序(定量)技术分析了对照样品和多效唑处理样品之间的基因表达谱,筛选出2838个差异表达基因。基因本体论(GO)、京都基因与基因组百科全书(KEGG)和MapMan通路分析表明,这些差异表达基因在对刺激的反应、激素信号传导、碳水化合物代谢、细胞壁、细胞大小和细胞周期相关生物过程中显著富集。为了验证通过RNA测序获得的表达谱,对从关键显著富集通路中选择的24个基因进行了实时定量PCR。综合分析表明,多效唑阻断赤霉素信号,可有效抑制花葶伸长;赤霉素信号与包括生长素、乙烯、油菜素内酯和细胞分裂素在内的其他激素信号相互作用,并触发下游信号级联反应,导致代谢、细胞壁生物合成、细胞分裂和细胞周期明显下降,最终诱导矮化性状。此外,AP2/EREBP、bHLH、C2H2、ARR、WRKY和ARF家族的转录因子参与了百子莲花葶发育的调控。这个转录组数据集将作为一个重要的公共信息平台,以加速对百子莲基因表达和功能基因组学的研究。