青花椒抗性基因ZaMBF1家族的生物信息学分析和逆境响应研究

doi:10.16036/j.issn.1000-2650.202301203

摘要/Abstract

摘要： 目的: 探究青花椒抗逆调控基因ZaMBF1基因家族特征和非生物胁迫下的表达模式,为青花椒抗性育种提供重要的基因来源和理论基础。方法: 结合近年发表的青花椒基因组数据,综合利用生物信息学研究方法和荧光定量PCR技术,解析了多蛋白侨联因子MBF1家族（multiprotein bridging factor 1）的核酸和蛋白序列特征,探究了ZaMBF1s对盐胁迫和脱落酸处理后的响应模式。结果: 青花椒共有3个MBF1家族成员,分别为ZaMBF1a、ZaMBF1b和ZaMBF1c。其中ZaMBF1a和ZaMBF1b编码序列长度均为429 bp,分别位于26号和32号染色体,而ZaMBF1c编码序列长度为453 bp,位于4号染色体;青花椒ZaMBF1s均为亲水性的不稳定蛋白,其核酸序列和对应的启动子序列均含有丰富的WRKY、bHLH等抗逆调控基因结合位点和顺式作用元件;ZaMBF1家族基因在NaCl外源处理3 h后即可快速响应,并在处理后12 h再次极显著上调,尤其是ZaMBF1c的上调最为显著;同时,外源处理12 h也是ZaMBF1家族基因响应ABA处理的重要时间点。结论: 青花椒ZaMBF1家族成员可以参与抗逆调控过程,并对外源盐和脱落酸处理具有显著响应。本研究所获结果对于探究ZaMBF1基因的分子功能,揭示青花椒抗逆性的调控机制提供了丰富的数据支撑。

关键词: 青花椒, 盐胁迫, 脱落酸处理, 多蛋白侨联因子

Abstract: 【Objective】 To explore the characters of ZaMBF1 gene family and its expression patterns in different abiotic stresses,and provide an important gene source and theoretical basis for resistance breeding in Zanthoxylum armatum. 【Method】 According to the public genome data of Z. armatum,this study comprehensively conduct the bioinformatics analysis of ZaMBF1 family （multiprotein bridging factor 1）,including nucleic acid and protein sequence characters,and the expression patterns of ZaMBF1s were also explored in response to salt stress and ABA treatment using quantitative reverse transcription PCR technology. 【Result】 A total of three MBF1 members were detected in Z. armatum,named ZaMBF1a,ZaMBF1b and ZaMBF1c. Both the coding sequences of ZaMBF1a and ZaMBF1b were 429 bp and located on chromosomes No. 26 and No. 32,respectively. However,the coding sequence of ZaMBF1c was 453 bp,located on chromosome No.4. Besides,all of the ZaMBF1 were belonged to hydrophilic and unstable proteins,and it was enriched a plenty of WRKY,bHLH and other stress-resistant regulatory binding sites and cis-acting elements in the ZaMBF1s nucleic acid sequences and the corresponding promoter sequences. Meanwhile,the ZaMBF1 genes rapidly responded at 3 h after exogenous NaCl treatment,and were significantly up-regulated at 12 h once again,especially the ZaMBF1c. Additionally,the 12 h after exogenous treatment was also an crucial time point for ZaMBF1 family genes in response to ABA treatment. 【Conclusion】 The ZaMBF1 family members could be involved in stress regulation,and respond significantly to exogenous salt and ABA treatment. The present results will provide enough support for exploring the molecular functions of ZaMBF1 genes and revealing the regulatory mechanism of stress resistance in Zanthoxylum armatum.

Key words: Zanthoxylum armatum, salt stress, ABA treatment, MBF1

中图分类号:

S718.3

王寒葶, 张雨, 李豪, 郑皓, 田源成, 朱舜棱, 许博洋, 王景燕, 龚伟, 惠文凯. 青花椒抗性基因ZaMBF1家族的生物信息学分析和逆境响应研究[J]. 四川农业大学学报, 2023, 41(4): 592-601.

WANG Hanting, ZHANG Yu, LI Hao, ZHENG Hao, TIAN Yuancheng, ZHU Shunling, XU Boyang, WANG Jingyan, GONG Wei, HUI Wenkai. Bioinformatics Analysis of the ZaMBF1 Family and Its Expression Pattern under Different Stresses in Zanthoxylum armatum[J]. Journal of Sichuan Agricultural University, 2023, 41(4): 592-601.

参考文献

[1] FENG S J, LIU Z S, HU Y, et al.Genomic analysis reveals the genetic diversity, population structure, evolutionary history and relationships of Chinese pepper[J]. Horticulture Research, 2020, 7: 158.
[2] DONG Z F, ZHAO X L, LIU J J, et al.Detection and simultaneous differentiation of three co-infected viruses in Zanthoxylum armatum[J]. Plants, 2022, 11(9): 1242.
[3] OKAGU I U, NDEFO J C, AHAM E C, et al.Zanthoxylum species: a comprehensive review of traditional uses, phytochemistry, pharmacological and nutraceutical applications[J]. Molecules (Basel, Switzerland), 2021, 26(13): 4023.
[4] LIU X M, HE X, LIU Z B, et al.Transcriptome mining of genes in Zanthoxylum armatum revealed ZaMYB86 as a negative regulator of prickly development[J]. Genomics, 2022, 114(3): 110374.
[5] HUI W K, WANG J Y, MA L X, et al.Identification of key genes in the biosynthesis pathways related to terpenoids, alkaloids and flavonoids in fruits of Zanthoxylum armatum[J]. Scientia Horticulturae, 2021, 290: 110523.
[6] KALIA N K, SINGH B, SOOD R P.A new amide from Zanthoxylum armatum[J]. Journal of Natural Products, 1999, 62(2): 311-312.
[7] FEI X T, HOU L X, SHI J W, et al.Patterns of drought response of 38 WRKY transcription factors of Zanthoxylum bungeanum maxim[J]. International Journal of Molecular Sciences, 2018, 20(1): 68.
[8] ARAVIND L, ANANNTHARAMAN V, BLALJI S, et al.Themany faces of the helixturn-helix domain: Transcription regulation and beyond[J].FEMS Microbiology Reviews, 2005, 29: 231-262.
[9] JAIMES-MIRANDA F, CHÁVEZ MONTES R A. The plant MBF₁ protein family: a bridge between stress and transcription[J]. Journal of Experimental Botany, 2020, 71(6): 1782-1791.
[10] TSUDA K, TSUJI T, HIROSE S, et al.Three Arabidopsis MBF₁ homologs with distinct expression profiles play roles as transcriptional co-activators[J]. Plant & Cell Physiology, 2004, 45(2): 225-231.
[11] HUANG S W, LIN Z Q, TUNG S Y, et al.A novel multiprotein bridging factor 1-like protein induces cyst wall protein gene expression and cyst differentiation in Giardia lamblia[J]. International Journal of Molecular Sciences, 2021, 22(3): 1370.
[12] HUI W K, ZHENG H, FAN J T, et al.Genome-wide characterization of the MBF₁ gene family and its expression pattern in different tissues and stresses in Zanthoxylum armatum[J]. BMC Genomics, 2022, 23(1): 652.
[13] LI F Q, UEDA H, HIROSE S, et al.Mediators of activation of fushi tarazu gene transcription by BmFTZ-F₁[J]. Molecular and Cellular Biology, 1994, 14(5): 3013-3021.
[14] GUO W L, CHEN R G, DU X H, et al.Reduced tolerance to abiotic stress in transgenic Arabidopsis overexpressing a Capsicum annuum multiprotein bridging factor 1[J]. BMC Plant Biology, 2014, 14(5): 138.
[15] TOJO T, TSUDA K, YOSHIZUMI T, et al.Arabidopsis MBF1s control leaf cell cycle and its expansion[J]. Plant & Cell Physiology, 2009, 50(2): 254-264.
[16] YU R M, SUO Y Y, YANG R, et al.StMBF1c positively regulates disease resistance to Ralstonia solanacearum via it's primary and secondary upregulation combining expression of StTPS5 and resistance marker genes in potato[J]. Plant Science: an International Journal of Experimental Plant Biology, 2021, 307: 110877.
[17] SUZUKI N, SEJIMA H, TAM R, et al.Identifcation of the MBF1 heat-response regulon of Arabidopsis thaliana[J]. Plant Journal: for Cell and Molecular Biology, 2011, 66(5): 844-851.
[18] NOBUHIRO S, LUDMILA R, LIANG H J, et al.Enhanced tolerance to environmental stress in transgenic plants expressing the transcriptional coactivator multiprotein bridging factor 1c[J]. Plant Physiology, 2005, 139(3): 1313-1322.
[19] LI M, HOU L, LIU S S, et al.Genome-wide identification and expression analysis of NAC transcription factors in Ziziphus jujuba Mill. reveal their putative regulatory effects on tissue senescence and abiotic stress responses[J]. Industrial Crops and Products, 2021, 173: 114093.
[20] 云雪雪, 陈雨生. 国际盐碱地开发动态及其对我国的启示[J]. 国土与自然资源研究, 2020(1): 84-87.
[21] 赵作章, 陈劲松, 彭尔瑞, 等. 土壤盐渍化及治理研究进展[J]. 中国农村水利水电, 2023(6): 202-208.
[22] JI X, CHENG J, GONG D H, et al.The effect of NaCl stress on photosynthetic efficiency and lipid production in freshwater microalga—Scenedesmus obliquus XJ002[J]. Science of the Total Environment, 2018, 633: 593-599.
[23] FEI X T, SHI Q Q, YANY T X, et al.Expression stabilities of ten candidate reference genes for RT-qPCR in Zanthoxylum bungeanum maxim[J]. Molecules, 2018, 23(4): 802.
[24] VYAS P, KAUR R.Culturable endophytic Pseudomonas fluorescens Z1B4 isolated from Zanthoxylum alatum Roxb. with stress-tolerance and plant growth-promoting potential[J]. BioTechnologia, 2021, 102(3): 285-295.
[25] 赵兰兰, 闻童, 赵兵, 等. 西南地区近50年干旱趋势及特征分析[J]. 水文, 2021, 41(6): 91-95, 59.
[26] 罗蛟, 李滢, 李玉婷, 等. 六种植物生长调节剂对光温胁迫下离体黄瓜叶片光系统Ⅱ和光系统Ⅰ光抑制的影响[J]. 植物生理学报, 2021, 57(1): 178-186.
[27] LUO X Y, LI C, HE X, et al.ABA signaling is negatively regulated by GbWRKY1 through JAZ1 and ABI1 to affect salt and drought tolerance[J]. Plant Cell Reports, 2020, 39(2): 181-194.
[28] WANG M C, TONG S F, MA T, et al.Chromosome‐level genome assembly of Sichuan pepper provides insights into apomixis, drought tolerance, and alkaloid biosynthesis[J]. Molecular Ecology Resources, 2021, 21(7): 2533-2545.
[29] ZHENG Y, JIAO C, SUN H H, et al.iTAK: a program for genome-wide prediction and\u00a0Classification of plant transcription factors, transcriptional regulators, and protein kinases[J]. Molecular Plant, 2016, 9: 1667-1670.
[30] SHEN W, LE S, LI Y, et al.SeqKit: a cross-platform and ultrafast toolkit for FASTA/Q file manipulation[J]. PLoS One, 2016, 11(10): e0163962.
[31] 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.
[32] CHEN C J, CHEN H, ZHANG Y, et al.TBtools-an integrative toolkit developed for interactive analyses of big biological data[J]. Molecular Plant, 2020, 13(18): 1194-1202.
[33] LESCOT M, DÉHAIS P, THIJS G, et al. 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.
[34] KIM B M, LEE H J, SONG Y H, et al.Effect of salt stress on the growth, mineral contents, and metabolite profiles of spinach[J]. Journal of the Science of Food and Agriculture, 2021, 101(9): 3787-3794.
[35] 隆春艳, 古洪辉, 汪正香, 等. 外源脱落酸对高温胁迫下菠菜光合与叶绿素荧光参数的影响[J]. 四川农业大学学报, 2017, 35(1): 24-30.
[36] BAKHT J, BANO A, SHAFI M, et al.Effect of abscisic acid applications on cold tolerance in chickpea (Cicer arietinum L.)[J]. European Journal of Agronomy, 2013, 44: 10-21.
[37] 冯举伶, 汪军成, 姚立蓉, 等. 大麦HvMBF1c克隆及其响应盐胁迫的表达模式研究[J]. 植物遗传资源学报, 2022, 23(4): 1175-1186.
[38] WU Z, LI T, ZHANG D H, et al.Lily HD-Zip I transcription factor LlHB16 promotes thermotolerance by activating LlHSFA2 and LlMBF1c[J]. Plant & Cell Physiol, 2022, 63(11):1792-1744.
[39] 王达菲, 陈国平, 赖文泉, 等. 新转录辅激活子多蛋白桥梁因子MBF1的研究进展[J]. 安徽农业科学, 2009, 37(26):12425-12427.
[40] ERPEN L, DEVI H S, GROSSER J W, et al.Potential use of the DREB/ERF, MYB, NAC and WRKY transcription factors to improve abiotic and biotic stress in transgenic plants[J]. Plant Cell, Tissue and Organ Culture (PCTOC), 2018, 132(1): 1-25.
[41] YANG H J, ZHOU Y, ZHANG Y N, et al.Identification of transcription factors of nitrate reductase gene promoters and NRE2 cis-element through yeast one-hybrid screening in Nicotiana tabacum[J]. BMC Plant Biology, 2019, 19(1): 145.
[42] 杨巍, 唐兵, 谭国飞, 等. 芥菜全基因组中Hsf基因家族鉴定与分析[J]. 四川农业大学学报, 2023, 41(2), 295-306.
[43] LI J R, CHEN X Z, ZHOU X X, et al.Identification of trihelix transcription factors in Pogostemon cablin reveals PatGT-1 negatively regulates patchoulol biosynthesis[J]. Industrial Crop and Products, 2021, 161: 113182.
[44] KHAKSAR G, TREESUBSUNTORN C, THIRAVETYAN P.Effect of exogenous methyl jasmonate on airborne benzene removal by Zamioculcas zamiifolia: the role of cytochrome P450 expression, salicylic acid, IAA, ROS and antioxidant activity[J]. Environmental and Experimental Botany, 2017, 138: 130-138.
[45] PANDEY SHREE P, SOMSSICH IMRE E.The role of WRKY transcription factors in plant immunity[J]. Plant Physiology, 2009, 150(4): 1648-1655.
[46] LI W X, PANG S Y, LU Z G, et al.Function and mechanism of WRKY transcription factors in abiotic stress responses of plants[J]. Plants, 2020, 9(11): 1515.
[47] QIN Y, MA X, YU G H, et al.Evolutionary history of trihelix family and their functional diversification[J]. DNA Research: an International Journal for Rapid Publication of Reports on Genes and Genomes, 2014, 21(5): 499-510.
[48] BENGOA LUONI S A, CENCI A, MOSCHEN S, et al. Genome-wide and comparative phylogenetic analysis of senescence-associated NAC transcription factors in sunflower (Helianthus annuus)[J]. BMC Genemics, 2021, 22(1): 893.
[49] FIGUEROA N, LODEYRO A F, CARRILLO N, et al.Meta-analysis reveals key features of the improved drought tolerance of plants overexpressing NAC transcription factors[J]. Environmental and Experimental Botany, 2021, 186: 104449.
[50] LU X Y, LIANG X Y, LI X, et al.Genome-wide characterisation and expression profiling of the LBD family in Salvia miltiorrhiza reveals the function of LBD50 in jasmonate signaling and phenolic biosynthesis[J]. Industrial Crop and Products, 2020, 144:112006.
[51] ZHAO Y, TIAN X J, WANG F, et al.Characterization of wheat MYB genes responsive to high temperatures[J]. BMC Plant Biology, 2017, 17(1): 208.