《植物生理学报》 2017, 53(8): 1468-1478

苹果抗寒矮化砧应答冷胁迫的转录因子基因鉴定

王海波, 程来亮, 何平, 常源升, 李林光^*

山东省果树研究所, 山东泰安271000

通信作者：李林光；E-mail: llg6536@163.com

摘　要：

利用RNA-seq鉴定抗寒苹果矮化砧木‘71-3-150’冷胁迫(4°C)响应过程中的转录因子。结果表明: 该砧木冷胁迫2、6、12和24 h时的样品与未处理相比, 差异表达的转录因子基因数分别为83、98、115和136, 共计228个, 表明大量转录因子参与该砧木的冷胁迫响应。这些转录因子基因属于32个家族, 占苹果转录因子总数的7.25%, 包括32个WRKY、30个AP2/ERF、28个bHLH、21个MYB、13个NAC、12个HSF、10个CO-like、9个MYB_related、8个TCP、6个bZIP、6个C2H2、6个C3H、6个GATA、5个G2-like、4个GRAS、4个HD-ZIP、4个NF-YC、4个ARF、3个Dof、3个Trihelix、2个GRF和2个NF-YB, 以及B3、DBB、E2F/DP、EIL、MIKC、M-type、SBP、TALE、YABBY和ZF-HD家族的各1个成员, 其中有8个在苹果参考基因组没有注释。这些基因总体上分为10类表达模式, 同一类表达模式中的基因均涉及到多个转录因子家族, 表明转录调控在该砧木应答冷胁迫中存在复杂机制。根据对鉴定到转录因子基因的功能分析, 它们参与了冷适应相关功能基因和生长发育相关功能基因的表达调控, 使该砧木在冷胁迫下重建体内物质代谢和氧化还原平衡, 获得抗寒性, 并且重建适应冷胁迫的新的生长与发育平衡。

关键词：苹果; 抗寒矮化砧; 冷胁迫; 转录因子

收稿：2016-12-31   修定：2017-07-17

资助：山东省农业科学院青年科研基金项目(2014QNZ04)、泰山学者建设工程专项经费(鲁政办字[2013]123号)、山东省农业科学院农业科技创新工程(CXGC2016B07)、山东省发展计划(2014GNC113004和2017GNC13113)和国家现代农业产业技术体系建设专项资金项目(CARS-28)。

Identification of genes encoding transcription factors of apple cold-resistant dwarfing rootstock in response to cold stress

WANG Hai-Bo, CHENG Lai-Liang, HE Ping, CHANG Yuan-Sheng, LI Lin-Guang^*

Shandong Institute of Pomology, Taian, Shandong 271000, China

Corresponding author: LI Lin-Guang; E-mail: llg6536@163.com

Abstract:

Using RNA-seq, the transcription factors (TFs) in ‘71-3-150’, a cold-resistant apple dwarf rootstock, responding to cold stress (4°C) of 0, 2, 6, 12 and 24 h were identified. In four comparison groups, 0 h vs 2 h, 0 h vs 6 h, 0 h vs 12 h, 0 h vs 24 h, 83, 98, 115 and 136 differentially expressed TF genes were found respectively. These total 228 TF genes, belonging to 32 families and comprising 7.25% of the total TFs of apple, were 32 WRKYs, 30 AP2/ERFs, 28 bHLHs, 21 MYBs, 13 NACs, 12 HSFs, 10 CO-like, 9 MYB_related, 8 TCPs, 6 bZIPs, 6 C2H2, 6 C3Hs, 6 GATAs, 5 G2-like, 4 GRAS, 4 HD-ZIPs, 4 NF-YCs, 4 ARFs, 3 Dofs, 3 Trihelix , 2 GRFs, 2 NF-YBs, 1 B3, 1 DBB, 1 E2F/DP, 1 EIL, 1 MIKC, 1 M-type, 1 SBP, 1 TALE, 1 YABBY, 1 ZF-HD, suggesting lots of TFs were involved in the rootstock responding to cold stress. Among them, 8 new genes without annotations in apple reference genome were identified. Ten differential expression clusters were obtained according to the trends of expression levels of all these genes. Various TF families were involved in each expression cluster indicating the complex mechanism of transcriptional regulation of the rootstock in response to cold stress. Based on functional analysis of indentified TF genes, they played the roles of regulating expression of functional genes in short-term cold acclimation and long-term growth and development, realizing the reestablishment of metabolites and redox balance to acquire cold tolerance, and new homeostasis of growth under cold stress.

Key words: apple; cold-resistant dwarfing rootstocks; cold stress; transcription factors