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作者中文名	作者英文名	单位中文名	单位英文名	E-Mail
李海燕	LI Hai-yan	沈阳农业大学园艺学院，辽宁沈阳 110866	Horticultural College, Shenyang Agricultural University, Shenyang 110866, China
刘天壤	LIU Tian-rang	沈阳农业大学园艺学院，辽宁沈阳 110866	Horticultural College, Shenyang Agricultural University, Shenyang 110866, China
陈佳惠	CHEN Jia-hui	沈阳农业大学园艺学院，辽宁沈阳 110866	Horticultural College, Shenyang Agricultural University, Shenyang 110866, China
王彪	WANG Biao	沈阳农业大学园艺学院，辽宁沈阳 110866	Horticultural College, Shenyang Agricultural University, Shenyang 110866, China
李宏博*	LI Hong-bo	沈阳农业大学园艺学院，辽宁沈阳 110866 道地药材国家重点实验室，北京 100700	Horticultural College, Shenyang Agricultural University, Shenyang 110866, China State Key Laboratory of Dao-di Herbs, Beijing 100700, China

基金项目:中央本级重大增减支项目（2060302）；2018年全国中药资源普查项目（1102-01042918001）

中文摘要:目的基于基因组数据鉴定丹参NAC家族转录因子（SmNACs）基因，并对其进行生物信息学与表达模式分析，为进一步深入阐明其基因功能提供依据。方法从丹参基因组数据库中鉴定出SmNACs基因，运用生物信息学方法及在线工具分析其结构特征，启动子顺式作用元件，编码蛋白的理化性质、亚细胞定位等，采用实时荧光定量聚合酶链式反应（qRT-PCR）测定SmNACs基因在丹参受到茉莉酸甲酯（MeJA）胁迫时的表达模式。结果从丹参的基因组中共确定了84个SmNACs基因（SmNAC01~SmNAC84），不均等分布于8条染色体上，编码蛋白的氨基酸长度为120~794 aa，等电点为4.27~10.28；基因结构显示SmNACs基因基本都含有1~10个内含子；上游启动子含有与激素、逆境胁迫应激相关的ABRE、MBS、ARE、LTR、CGTCA-motif、TGACG-motif等顺式作用元件；构建丹参、南丹参与拟南芥的NAC家族成员的系统发育树，将84个SmNACs基因分为23个亚族。转录组数据显示SmNACs基因在丹参不同组织中差异表达，其中7个SmNACs基因在根中的表达水平较高；MeJA处理后，17个SmNACs基因表达水平明显上调，20个SmNACs基因表达水平明显下调。qRT-PCR表达分析发现，MeJA处理后，12个SmNACs基因的表达模式随着激素处理时间的增加呈现上调或下调的趋势，SmNAC09、SmNAC15、SmNAC16受MeJA诱导后48 h表达水平显著上调，SmNAC20、SmNAC77、SmNAC78受MeJA诱导表达水平下调。结论研究结果为进一步解析SmNACs基因在丹参逆境响应及次生代谢产物生物合成中的调控机制提供了参考。

中文关键词:丹参基因组 NAC转录因子生物信息学茉莉酸甲酯表达分析

Genome-wide Identification and Expression Analysis of NAC Gene Family in Salvia miltiorrhiza

Abstract:Objective To identify NAC transcription factors of Salvia miltiorrhiza (SmNACs) and analyze bioinformatics and expression patterns based on genomic data to provide a basis for further elucidating the gene functions.Methods NAC family genes were identified based on the S. miltiorrhiza genome database, and the gene structure, promoter cis-elements, the physical and chemical properties of the encoded proteins, and subcellular localization were analyzed using bioinformatics tools. In addition, expression patterns of SmNACs in S. miltiorrhiza under methyl jasmonate (MeJA) stress were determined by the quantitative real-time polymerase chain reaction (qRT-PCR).Results Eighty-four NAC transcription factors (SmNAC01-SmNAC84) were identified from the genome of S. miltiorrhiza which were distributed unequally on the eight chromosomes, and the SmNACs consisted of 120-794 amino acids (aa) with the isoelectric point in the range of 4.27 to 10.28. The gene structures of those SmNACs showed that their introns varied from one to ten. The upstream promoters contained cis-acting elements such as ABRE, MBS, ARE, LTR, CGTCA-motif, and TGACG-motif related to hormones and stresses. Phylogenetic analysis divided the NAC gene family from S. miltiorrhiza, S. bowleyana, and Arabidopsis thaliana into 23 subgroups. Transcriptome data indicated that SmNACs were differentially expressed in different tissues of S. miltiorrhiza, among them, seven SmNACs were highly expressed in roots. Under MeJA treatment, 17 SmNACs were significantly up-regulated, and 20 SmNACs were significantly down-regulated. As revealed by qRT-PCR, 12 SmNACs were differentially expressed after MeJA treatment, and specifically, SmNAC09, SmNAC15, and SmNAC16 were significantly up-regulated 48 h after MeJA treatment, while SmNAC20, SmNAC77, and SmNAC78 were significantly down-regulated.Conclusion This study is expected to provide a theoretical foundation for further analysis of the regulatory mechanism of SmNACs in the stress response and secondary metabolite biosynthesis of S. miltiorrhiza.

keywords:Salvia miltiorrhiza Bunge genome NAC transcription factors bioinformatics MeJA expression patterns

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