中国农业科学 ›› 2016, Vol. 49 ›› Issue (7): 1325-1345.doi: 10.3864/j.issn.0578-1752.2016.07.010

• 园艺 • 上一篇    下一篇

苹果bZIP转录因子家族生物信息学分析及其在休眠芽中的表达

孙明岳,周 君,谭秋平,付喜玲,陈修德,李 玲,高东升   

  1. 山东农业大学园艺科学与工程学院/国家苹果工程技术研究中心/作物生物学国家重点实验室/山东果蔬优质高效生产协同创新中心,山东泰安 271018
  • 收稿日期:2015-11-26 出版日期:2016-04-01 发布日期:2016-04-01
  • 通讯作者: 高东升,Tel:0538-8249659;E-mail:dsgao@sdau.edu.cn;李玲,Tel:0538-8246168;E-mail:lilingsdau@163.com
  • 作者简介:孙明岳,Tel:0538-8249659;E-mail:smysdau@163.com
  • 基金资助:
    国家自然科学基金(31372050)、山东省自然科学基金(ZR2014CM015)

Analysis of Basic Leucine Zipper Genes and Their Expression During Bud Dormancy in Apple (Malus×domestica)

SUN Ming-yue, ZHOU Jun, TAN Qiu-ping, FU Xi-ling, CHEN Xiu-de, LI Ling, GAO Dong-sheng   

  1. College of Horticulture Science and Engineering, Shandong Agricultural University/National Research Center for Apple Engineering and Technology/State Key Laboratory of Crop Biology/Shandong Collaborative Innovation Center for Fruit and Vegetable Production with High Quality and Efficiency, Taian 271018, Shandong
  • Received:2015-11-26 Online:2016-04-01 Published:2016-04-01

摘要: 【目的】鉴定苹果(Malus×domestica Borkh.)基因组上的bZIP基因(MdbZIP),为研究苹果bZIP转录因子提供相关信息以及在芽休眠过程中的调控作用提供理论参考。【方法】通过Pfam下载bZIP隐马尔科夫模型bZIP_1(PF00170)与bZIP_2(PF07716),利用HMMER 3.0鉴定苹果bZIP基因。使用Clustal Omega、MEGA6.0、MapInspect、DNAMAN 6.0和MEME4.10.2等软件对其蛋白序列进行生物信息学分析。采用Microarray分析与qRT-PCR技术检测苹果bZIP基因在不同处理下及其在高需冷量品种与低需冷量品种中的表达情况。【结果】鉴定得到120个苹果bZIP基因,与拟南芥的系统进化树分析将苹果bZIP分为10个亚家族(A—I和S)。染色体定位分析显示,109个苹果bZIP不均匀分布于17条染色体上,其中,11个基因无匹配的染色体定位。8号染色体上分布最多(13个),1号染色体分布最少(1个),一些染色体区域基因密度较高。基因结构分析表明,MdbZIP基因家族外显子数量0—23个,其中23个基因无内含子,分布于F亚家族(4)与S亚家族(19),基因结构进化高度保守。保守元件分析表明,MdbZIP基因家族包含30个保守元件:元件1为bZIP保守结构域;在D亚家族发现的元件10与G亚家族发现的14为已知元件,另外多数元件功能未知。通过Microarray分析显示,多个MdbZIP均可能与芽休眠的解除相关。qRT-PCR结果显示在不同品种中A亚家族8个MdbZIP均呈现出ABA诱导表达,而D亚家族中随着冷处理时间延长,在需冷量不同的品种中出现多种表达模式。【结论】苹果bZIP基因家族结构高度保守,在ABA与冷处理下呈现不同表达模式,可能参与调控苹果芽休眠进程。

关键词: 苹果, bZIP转录因子, 芽休眠, 进化树分析, 序列对比, Microarray分析, qRT-PCR

Abstract: 【Objective】In this study, putative bZIP transcription factor-encoding genes in the apple (Malus×domestica Borkh.) genome were identified, so as to provide a basis for studying the theoretical roles of bZIP genes in the bud dormancy, and to provide valuable information for bZIP genes in apple. 【Method】 The Hidden Markov Model profiles of the bZIP domains (PF00170 and PF07716) downloaded from Pfam were used to search the database using HMMER, ver. 3.0 with the default E-value. The obtained amino acid sequences were analyzed with the bioinformatics softwares, including Clustal Omega, MEGA6.0, MapInspect, DNAMAN 6.0 and MEME4.10.2. Furthermore, Microarray analysis and qRT-PCR results indicated that many MdbZIP genes may be involved in regulating bud dormancy in different cultivar of apples. 【Result】 In Total 120 MdbZIP genes were found in the apple genome. Phylogenetic analyses of the genes based on A. thaliana counterparts indicated that the transcription factors can be categorized into 10 different groups (Groups A–I and S), Chromosome mapping analysis showed that 109 MdbZIP genes were distributed unevenly on 17 chromosomes. Except for 11 genes that were not located on chromosomes, the largest number of MdbZIP genes were found on chromosomes 8 (thirteen genes), only 1 genes located on chromosome 1, some chromosomal regions had a relatively high density of MdbZIP genes. The results of gene structure analysis revealed that MdbZIP gene contained 0-23 exons, 23 bZIP genes were intronless and were found in Groups S (nine genes) and F (one gene), bZIP gene structure were highly conserved in apples. Conserved motif analysis showed that the conserved motifs 1, which specify the bZIP domain were observed in all apple bZIP proteins, motif 10 and 14 as the known domain were observed in Group D and G, respectively. Furthermore, microarray data indicated that many MdbZIP genes may be involved in regulating bud dormancy release. And qRT-PCR results indicated that ABA induce eight members of group A expression in high-chill and low-chill apples, but the largest number of differentially expressed genes was found in group D with cold temperatures. 【Conclusion】 These results suggested that MdbZIP gene family was highly and structurally conserved, and involved in abscisic acid (ABA) signaling and cold stresses thereby possibly involved into the regulation of bud dormancy in apples.

Key words: apple (Malus×domestica Borkh.) , bZIP transcription factor, bud dormancy, phylogenetic analysis, sequence alignment, microarray analysis, qRT-PCR