State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, 51 Hexing Road, Harbin, 150040, China.
Co-Innovation Center for Sustainable Forestry in Southern China/Bamboo Research Institute, Nanjing Forestry University, 159 Longpan Road, Nanjing, 210037, China.
BMC Plant Biol. 2020 Jan 8;20(1):12. doi: 10.1186/s12870-019-2191-2.
NAC (NAM/ATAF/CUC) is one of the largest plant-specific transcription factor (TF) families known to play significant roles in wood formation. Acting as master gene regulators, a few NAC genes can activate secondary wall biosynthesis during wood formation in woody plants.
In the present study, firstly, we screened 110 differentially expressed NAC genes in the leaves, stems, and roots of di-haploid Populus simonii×P. nigra by RNA-Seq. Then we identified a nucleus-targeted gene, NAC15 gene, which was one of the highly expressed genes in the stem among 110 NAC family members. Thirdly, we conducted expression pattern analysis of NAC15 gene, and observed NAC15 gene was most highly expressed in the xylem by RT-qPCR. Moreover, we transferred NAC15 gene into tobacco and obtained 12 transgenic lines overexpressing NAC15 gene (TLs). And the relative higher content of hemicellulose, cellulose and lignin was observed in the TLs compared to the control lines containing empty vector (CLs). It also showed darker staining in the culms of the TLs with phloroglucinol staining, compared to the CLs. Furthermore, the relative expression level of a few lignin- and cellulose-related genes was significantly higher in the TLs than that in the CLs.
The overall results indicated that NAC15 gene is highly expressed in the xylem of poplar and may be a potential candidate gene playing an important role in wood formation in transgenic tobacco.
NAC(NAM/ATAF/CUC)是已知在植物中发挥重要作用的最大的一类特定于植物的转录因子(TF)家族之一,在木质部形成中。作为主基因调控因子,少数 NAC 基因可以在木本植物木质部形成过程中激活次生壁生物合成。
在本研究中,我们首先通过 RNA-Seq 筛选了二倍体杨树叶片、茎和根中的 110 个差异表达的 NAC 基因。然后,我们鉴定了一个核靶向基因 NAC15 基因,它是 110 个 NAC 家族成员中茎中高表达的基因之一。第三,我们对 NAC15 基因的表达模式进行了分析,观察到 NAC15 基因在木质部中表达最高,通过 RT-qPCR 验证。此外,我们将 NAC15 基因转入烟草,获得了 12 个过表达 NAC15 基因的转基因株系(TLs)。与含有空载体的对照株系(CLs)相比,TLs 中的半纤维素、纤维素和木质素含量相对较高。与 CLs 相比,TLs 的木质部用间苯三酚染色后颜色更深。此外,TLs 中一些木质素和纤维素相关基因的相对表达水平明显高于 CLs。
总体结果表明,NAC15 基因在杨树的木质部中高度表达,可能是一个在转基因烟草木质部形成中起重要作用的潜在候选基因。