苹果多胺氧化酶(PAO)基因家族鉴定与表达分析

doi:10.3969/j.issn.1004-1524.2020.02.10

浙江农业学报 ›› 2020, Vol. 32 ›› Issue (2): 262-273.DOI: 10.3969/j.issn.1004-1524.2020.02.10

苹果多胺氧化酶(PAO)基因家族鉴定与表达分析

秦玲¹, 张鑫¹, 荣春笑¹, 莫传园¹, 范露¹, 闫婕¹, 孟莹², 张满让^1,*

1.西北农林科技大学园艺学院,陕西杨凌 712100;
2.洛南县林业局,陕西商洛 726100

收稿日期:2019-07-11 出版日期:2020-02-25 发布日期:2020-03-13
通讯作者: *张满让,E-mail:mra@nwsuaf.edu.cn
作者简介:秦玲(1994—),女,四川遂宁人,硕士,研究方向为果树生理生态。E-mail:875332955@qq.com
基金资助:
国家现代农业产业技术体系(CARS-27)

Identification and expression analysis of polyamine oxidase (PAO) gene family in apple

QIN Ling¹, ZHANG Xin¹, RONG Chunxiao¹, MO Chuanyuan¹, FAN Lu¹, YAN Jie¹, MENG Ying², ZHANG Manrang^1,*

1. College of Horticulture, Northwest A&F University, Yangling 712100, China;
2. Forestry Bureau of Luonan County, Luonan 726100, China

Received:2019-07-11 Online:2020-02-25 Published:2020-03-13

摘要/Abstract

摘要： 从金冠基因组数据库中,利用BLAST网站在苹果中鉴定得到8个编码多胺氧化酶(polyamine oxidase,PAO)的家族基因(命名为MdPAO1~8),对其进行理化性质、系统进化、蛋白结构、基因结构和启动子元件分析。结果表明,MdPAO蛋白的分子量为54.053~64.813 ku,等电点介于5.16~6.02。根据进化树分析,MdPAO被分为3个亚家族(Ⅰ、Ⅲ、Ⅳ),且同一亚家族的成员具有相似的蛋白结构、基因结构和蛋白保守基序分布。利用GEO数据库检测出8个MdPAO在不同组织器官中的表达具有明显差异。以苹果主栽品种长富2号为材料,利用实时荧光定量PCR检测分析得出8个MdPAO在不同组织中的表达情况,结果表明,MdPAO3、MdPAO4、MdPAO5/6和MdPAO8在花中有较高的表达水平;MdPAO1/2和MdPAO7在茎和果中有较高的表达水平。外源亚精胺处理后发现,除MdPAO3以外,其余MdPAO的转录水平显著提高,表明MdPAO在PA代谢途径中具有重要作用。

关键词: 苹果, 多胺氧化酶(PAO)基因家族, 生物信息学, 基因表达

Abstract: From the genomic database of Golden Delicious, eight family genes encoding polyamine oxidase (PAO) (named as MdPAO1-8) were identified in apples using the BLAST website for physicochemical properties, phylogenetic, protein structure, analysis of gene structure and promoter elements. The results showed that the molecular weight of MdPAO protein ranged from 54.053 to 64.813 ku, and the isoelectric point ranged from 5.16 to 6.02. According to the phylogenetic tree analysis, MdPAO is divided into three subfamilies (Ⅰ,Ⅲ, Ⅳ), and members of the same subfamily share similar protein structure, gene structure and protein conserved motif distribution. The expression of 8 MdPAO genes in different tissues and organs was significantly different using the GEO database. The expression of 8 MdPAO genes in different tissues was determined by real-time quantitative PCR using Nagafu No.2. The results showed that MdPAO3, MdPAO4, MdPAO5/6 and MdPAO8 were highly espressed in flowers. MdPAO1/2 and MdPAO7 had higher expression levels in stems and fruits. Exogenous spermidine treatment showed that the transcriptional level of MdPAO significantly increased except MdPAO3, indicating that MdPAO plays an important role in the PA metabolic pathway.

Key words: apple, polyamine oxidase (PAO) gene family, bioinformatics, gene expression

中图分类号:

S661.1

秦玲, 张鑫, 荣春笑, 莫传园, 范露, 闫婕, 孟莹, 张满让. 苹果多胺氧化酶(PAO)基因家族鉴定与表达分析[J]. 浙江农业学报, 2020, 32(2): 262-273.

QIN Ling, ZHANG Xin, RONG Chunxiao, MO Chuanyuan, FAN Lu, YAN Jie, MENG Ying, ZHANG Manrang. Identification and expression analysis of polyamine oxidase (PAO) gene family in apple[J]. , 2020, 32(2): 262-273.

参考文献

[1] ALCÁZAR R, ALTABELLA T, MARCO F, et al. Polyamines: molecules with regulatory functions in plant abiotic stress tolerance[J]. Planta, 2010, 231(6): 1237-1249.
[2] AHMED S, ARIYARATNE M, PATEL J, et al.Altered expression of polyamine transporters reveals a role for spermidine in the timing of flowering and other developmental response pathways[J]. Plant Science, 2017, 258: 146-155.
[3] PLANAS-PORTELL J, GALLART M, TIBURCIO A F, et al.Copper-containing amine oxidases contribute to terminal polyamine oxidation in peroxisomes and apoplast of Arabidopsis thaliana[J]. BMC Plant Biology, 2013, 13(1): 109.
[4] MOSCHOU P N, WU J, CONA A, et al.The polyamines and their catabolic products are significant players in the turnover of nitrogenous molecules in plants[J]. Journal of Experimental Botany, 2012, 63(14): 5003-5015.
[5] AHOU A, MARTIGNAGO D, ALABDALLAH O, et al.A plant spermine oxidase/dehydrogenase regulated by the proteasome and polyamines[J]. Journal of Experimental Botany, 2014, 65(6): 1585-1603.
[6] FINCATO P, MOSCHOU P N, SPEDALETTI V, et al.Functional diversity inside the Arabidopsis polyamine oxidase gene family[J]. Journal of Experimental Botany, 2011, 62(3): 1155-1168.
[7] ONO Y, KIM D W, WATANABE K, et al.Constitutively and highly expressed Oryza sativa polyamine oxidases localize in peroxisomes and catalyze polyamine back conversion[J]. Amino Acids, 2012, 42(2/3): 867-876.
[8] CERVELLI M, CONA A, ANGELINI R, et al.A barley polyamine oxidase isoform with distinct structural features and subcellular localization[J]. European Journal of Biochemistry, 2001, 268(13): 3816-3830.
[9] CERVELLI M, TAVLADORAKI P, DI AGOSTINO S, et al.Isolation and characterization of three polyamine oxidase genes from Zea mays[J]. Plant Physiology and Biochemistry, 2000, 38(9): 667-677.
[10] YODA H, HIROI Y, SANO H.Polyamine oxidase is one of the key elements for oxidative burst to induce programmed cell death in tobacco cultured cells[J]. Plant Physiology, 2006, 142(1): 193-206.
[11] TUSKAN G A, DIFAZIO S, JANSSON S, et al.The genome of black cottonwood, Populus trichocarpa(Torr. & Gray)[J]. Science, 2006, 313: 1596-1604.
[12] WANG W, LIU J H.Genome-wide identification and expression analysis of the polyamine oxidase gene family in sweet orange (Citrus sinensis)[J]. Gene, 2015, 555(2): 421-429.
[13] CHENG X Q, ZHU X F, TIAN W G, et al.Genome-wide identification and expression analysis of polyamine oxidase genes in upland cotton (Gossypium hirsutum L.)[J]. Plant Cell, Tissue and Organ Culture, 2017, 129(2): 237-249.
[14] HAO Y W, HUANG B B, JIA D Y, et al.Identification of seven polyamine oxidase genes in tomato (Solanum lycopersicum L.) and their expression profiles under physiological and various stress conditions[J]. Journal of Plant Physiology, 2018, 228: 1-11.
[15] KITASHIBA H, HONDA C, MORIGUCHI T.Identification of polyamine oxidase genes from apple and expression analysis during fruit development and cell growth[J]. Plant Biotechnology, 2006, 23: 425-429.
[16] SAGOR G H M, INOUE M, KIM D W, et al. The polyamine oxidase from lycophyte Selaginella lepidophylla(SelPAO5), unlike that of angiosperms, back-converts thermospermine to norspermidine[J]. FEBS Letters, 2015, 589(20PartB): 3071-3078.
[17] WANG W, PASCHALIDIS K, FENG J C, et al.Polyamine catabolism in plants: a universal process with diverse functions[J]. Frontiers in Plant Science, 2019, 10: 561.
[18] KIM D W, WATANABE K, MURAYAMA C, et al.Polyamine Oxidase5 regulates Arabidopsis growth through thermospermine oxidase activity[J]. Plant Physiology, 2014, 165(4): 1575-1590.
[19] SAGOR G, KUSANO T, BERBERICH T.Identification of the actual coding region for polyamine oxidase 6 from rice (OsPAO6) and its partial characterization[J]. Plant Signaling & Behavior, 2017, 12(8): e1359456.
[20] CERVELLI M, BIANCHI M, CONA A, et al.Barley polyamine oxidase isoforms 1 and 2, a peculiar case of gene duplication[J]. FEBS Journal, 2006, 273(17): 3990-4002.
[21] KASUKABE Y, HE L X, NADA K, et al.Overexpression of spermidine synthase enhances tolerance to multiple environmental stresses and up-regulates the expression of various stress-regulated genes in transgenic Arabidopsis thaliana[J]. Plant and Cell Physiology, 2004, 45(6): 712-722.
[22] MOSCHOU P N, SANMARTIN M, ANDRIOPOULOU A H, et al.Bridging the gap between plant and mammalian polyamine catabolism: a novel peroxisomal polyamine oxidase responsible for a full back-conversion pathway in Arabidopsis[J]. Plant Physiology, 2008, 147: 1845-1857.
[23] SHEN W Y, NADA K, TACHIBANA S.Involvement of polyamines in the chilling tolerance of cucumber cultivars[J]. Plant Physiology, 2000, 124(1): 431-440.
[24] 曹尚银,张俊昌,江爱华,等. 苹果花芽孕育调控的最佳时期研究[J]. 中国果树, 2001(1): 14-17.
CAO S Y, ZHANG J C, JIANG A H, et al.Study on the best period of regulation of apple flower buds[J]. China Fruits,2001 (1):14-17. (in Chinese)
[25] FAN S, ZHANG D, XING L B, et al.Phylogenetic analysis of IDD gene family and characterization of its expression in response to flower induction in Malus[J]. Molecular Genetics and Genomics, 2017, 292(4): 755-771.
[26] LI Y M, ZHANG D, ZHANG X, et al.A transcriptome analysis of two apple (Malus×domestica) cultivars with different flowering abilities reveals a gene network module associated with floral transitions[J]. Scientia Horticulturae, 2018, 239: 269-281.
[27] 许园园, 蔺经, 李晓刚, 等. 梨CBL基因家族全基因组序列的鉴定及非生物胁迫下的表达分析[J]. 中国农业科学, 2015, 48(4): 735-747.
XU Y Y, LIN J, LI X G, et al.Identification and expression analysis under abiotic stresses of the CBL gene family in pear[J]. Scientia Agricultura Sinica, 2015, 48(4): 735-747.(in Chinese with English abstract)
[28] ARTIMO P, JONNALAGEDDA M, ARNOLD K, et al.ExPASy: SIB bioinformatics resource portal[J]. Nucleic Acids Research, 2012, 40(W1): W597-W603.
[29] ZHANG H X, JIN J H, HE Y M, et al.Genome-wide identification and analysis of the SBP-box family genes under Phytophthora capsici stress in pepper (Capsicum annuum L.)[J]. Frontiers in Plant Science, 2016, 7: 504.
[30] TAMURA K, STECHER G, PETERSON D, et al.MEGA6: molecular evolutionary genetics analysis version 6.0[J]. Molecular Biology and Evolution, 2013, 30(12): 2725-2729.
[31] GEOURJON C, DELÉAGE G. SOPMA: significant improvements in protein secondary structure prediction by consensus prediction from multiple alignments[J]. Computer Applications in the Biosciences: CABIOS, 1995, 11(6): 681-684.
[32] KELLEY L A, MEZULIS S, YATES C M, et al.The Phyre2 web portal for protein modeling, prediction and analysis[J]. Nature Protocols, 2015, 10(6): 845-858.
[33] HORTON P, PARK K J, OBAYASHI T, et al.WoLF PSORT: protein localization predictor[J]. Nucleic Acids Research, 2007, 35: W585-W587.
[34] HU B, JIN J P, GUO A Y, et al.GSDS 2.0: an upgraded gene feature visualization server[J]. Bioinformatics, 2015, 31(8): 1296-1297.
[35] BAILEY T L, WILLIAMS N, MISLEH C, et al.MEME: discovering and analyzing DNA and protein sequence motifs[J]. Nucleic Acids Research, 2006, 34(Web Server): W369-W373.
[36] LESCOT M.PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences[J]. Nucleic Acids Research, 2002, 30(1): 325-327.
[37] LI J, HOU H M, LI X Q, et al.Genome-wide identification and analysis of the SBP-box family genes in apple (Malus × domestica Borkh.)[J]. Plant Physiology and Biochemistry, 2013, 70: 100-114.
[38] GAMBINO G, PERRONE I, GRIBAUDO I.A rapid and effective method for RNA extraction from different tissues of grapevine and other woody plants[J]. Phytochemical Analysis, 2008, 19(6): 520-525.
[39] FAN S, ZHANG D, LEI C, et al.Proteome analyses using iTRAQ labeling reveal critical mechanisms in alternate bearing Malus prunifolia[J]. Journal of Proteome Research, 2016, 15(10): 3602-3616.
[40] 樊胜, 张岚清, 刘柯, 等. 苹果‘长富2号’开花调控转录因子基因MdSPL6的克隆及表达分析[J]. 园艺学报, 2016, 43(11): 2089-2098.
FAN S, ZHANG L Q, LIU K, et al.Cloning and expression of the flowering regulation transcription factor gene MdSPL6 in Malus × domestica[J]. Acta Horticulturae Sinica, 2016, 43(11): 2089-2098.(in Chinese with English abstract)
[41] LIVAK K J, SCHMITTGEN T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2^-ΔΔCT method[J]. Methods, 2001, 25(4): 402-408.
[42] BARRETT T, WILHITE S E, LEDOUX P, et al.NCBI GEO: archive for functional genomics data sets-update[J]. Nucleic Acids Research, 2013, 41(D1): D991-D995.
[43] WU T Y, YANKOVSKAYA V, MCINTIRE W S.Cloning, sequencing, and heterologous expression of the murine peroxisomal flavoprotein, N1-acetylated polyamine oxidase[J]. Journal of Biological Chemistry, 2003, 278(23): 20514-20525.
[44] KAMADA-NOBUSADA T, HAYASHI M, FUKAZAWA M, et al.A putative peroxisomal polyamine oxidase, AtPAO4, is involved in polyamine catabolism in Arabidopsis thaliana[J]. Plant and Cell Physiology, 2008, 49(9): 1272-1282.
[45] 邵红霞, 陈鸿飞, 张东, 等. 苹果YABBY基因家族的鉴定、进化及表达分析[J]. 浙江农业学报, 2017, 29(7): 1129-1138.
SHAO H X, CHEN H F, ZHANG D, et al.Identification, evolution and expression analysis of the YABBY gene family in apple (Malus×domestica Borkh.)[J]. Acta Agriculturae Zhejiangensis, 2017, 29(7): 1129-1138.(in Chinese with English abstract)
[46] VELASCO R, ZHARKIKH A, AFFOURTIT J, et al.The genome of the domesticated apple (Malus×domestica Borkh.)[J]. Nature Genetics, 2010, 42(10): 833.

苹果多胺氧化酶(PAO)基因家族鉴定与表达分析

Identification and expression analysis of polyamine oxidase (PAO) gene family in apple

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