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红豆杉(Taxus spp.)是举世公认的珍贵林木,在全世界一共有11种,我国存在4种和1变种,即东北红豆杉(T. cuspidata Sieb. et Zucc.)、西藏红豆杉(T. wallichiana Zucc.)、云南红豆杉(T. yunnanensis Cheng et L. K. Fu)、中国红豆杉(T. chinensis (Pilger) Rehd.)以及南方红豆杉(T. chinensis var. mairei (Lemeee et Levl) Cheng et L. K. Fu)[1]。曼地亚红豆杉(Taxus × media Rehder)为一种天然杂交种,于上世纪90年代由美国或加拿大引种于我国。
NAC (NAM/ATAF/CUC)是植物中最常见的转录因子家族之一,在植物生长发育过程中起到重要的调控作用,如响应非生物胁迫、调控器官生长发育以及参与激素信号转导等[2-4]。研究发现,水稻OsNAC2蛋白能够通过影响植物生长素和细胞分裂素(cytokinin)反应基因来调控根的发育[5], 而拟南芥ANAC092可以与生长素应答因子ARF8和PIN4的启动子相结合,通过控制生长素信号途径来影响拟南芥根系的发育[6-8]。此外,在拟南芥中异源表达BnNAC14、GmNAC109、CiNAC3和CiNAC4等其他物种的NAC基因,也可以对侧根的形成起到促进作用[8-11]。这些研究结果表明,NAC转录因子在植物根系的形成过程中扮演重要角色。
近年来,人们为了缓解红豆杉开发与利用的矛盾,在红豆杉人工种植研究上取得了巨大进展,但仍存在红豆杉扦插繁殖生根较慢(大约需要45~60 d)的技术瓶颈。并且,目前关于红豆杉生根的分子机制研究也较为少见。本研究从曼地亚红豆杉三代全长转录组数据中鉴定出了44个NAC转录因子基因,并对该基因家族进行了系统生物信息学及组织表达分析,为探索NAC转录因子调控红豆杉根系的形成奠定理论基础,有助于今后利用分子手段调控其根系生长、缩短扦插育苗时间。
曼地亚红豆杉NAC基因家族鉴定及表达分析
Identification and Expression Analysis of NAC Gene Family in Taxus × media Rehder
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摘要:
目的 筛选红豆杉NAC转录因子家族中影响根系形成的关键基因,探索红豆杉根系生长发育分子机理。 方法 利用曼地亚红豆杉全长转录组数据,通过生物信息学方法鉴定NAC 转录因子,并对筛选出的NAC 转录因子进行蛋白结构及基因组织表达谱等分析。 结果 共鉴定出 44个NAC 转录因子,其在N端均具有典型的保守NAC结构域,分别聚类到拟南芥的7个亚家族中,蛋白三级结构较为相似,且成员大多含有5个保守亚结构域。组织表达谱显示TmNAC15、16、18、21、22、29、39、40、41和44在根中表达水平高于茎和叶。 结论 从曼地亚红豆杉中共鉴定出44个NAC 转录因子,其结构较为保守,且聚类为7个亚家族,其中成员TmNAC21、22、39、40和44极有可能参与红豆杉根系生长发育。 Abstract:Objective To obtain the key NAC genes involving in regulating the root growth and development of yew trees. Method Basing on the full-length transcriptome data of Taxus × media, bioinformatics methods were used to identify NAC transcription factors. The protein structure and gene tissue expression profile of the selected NAC genes were also analyzed. Result A total of 44 NAC transcription factors were identified, which have typical conserved NAC domains at the N-terminal, and most of the members contain 5 conserved structural functional subdomains. Phylogenetic tree analysis showed that 44 T. × media NAC (TmNAC) proteins were clustered into 7 subfamilies of Arabidopsis. Sequence structure analysis indicated that the tertiary structures of the TmNAC proteins were similar, but the structures of the representative member of different subfamily also showed obvious differences. The expression results showed that TmNAC15, 16, 18, 21, 22, 29, 39, 40, 41, and 44 had higher expression level in roots than stems and needles. Conclusion In this study, 44 TmNAC transcription factors were identified from T. × media and they were clustered into 7 subfamilies. Among them, TmNAC21, 22, 39, 40, and 44 may be involved in the root growth and development progresses of yew trees. -
Key words:
- Taxus spp.
- / NAC gene
- / gene identification
- / expression analysis
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