桃PpNAC72的克隆及表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2018-0891

生物技术通报 ›› 2019, Vol. 35 ›› Issue (6): 16-23.doi: 10.13560/j.cnki.biotech.bull.1985.2018-0891

桃PpNAC72的克隆及表达分析

武肖琦, 冯涛, 刘艳军, 焦思鹏, 孙攀, 杨静慧

天津农学院园艺学院,天津 300384

收稿日期:2018-10-17 出版日期:2019-06-26 发布日期:2019-07-08
作者简介:武肖琦,女,硕士,研究方向：果树分子生物学;E-mail：1225053239@qq.com
基金资助:
天津农学院研究生创新培育项目（J0100906）,天津市科委项目（16YFZCNC00750）,中央财政项目（天津市重大农业技术推广项目）（2017CK0184）,天津市农委项目（ITTHRS2018002）,天津市科委特派员项目（17ZXBFNC00310）,天津市高等学校科技发展基金计划（20130617）,天津市林果现代产业技术体系（ITTFPRS2018004）

Cloning and Expression Analysis of the PpNAC72 in Peach

WU Xiao-qi, FENG Tao, LIU Yan-jun, JIAO Si-peng, SUN Pan, YANG Jing-hui

College of Horticulture,Tianjin Agricultural University,Tianjin 300384

Received:2018-10-17 Published:2019-06-26 Online:2019-07-08

摘要/Abstract

摘要： 旨在从桃树‘津柳早红’中克隆PpNAC72,并分析其在不同发育时期的表达情况,初步探究该基因在桃生长中的功能。采用RT-PCR技术从桃果实中克隆得到PpNAC72序列,并对其进行定量表达分析以及生物信息学分析。结果显示,桃PpNAC72（GenBank：MH627940）开放阅读框序列为1 050 bp,编码349个氨基酸,其等电点为8.34,有1个NAM结构域,具有典型的NAC家族结构特征。氨基酸同源性分析表明,PpNAC72与已报道的其他植物NAC蛋白具有较高的相似性,其中,与甜樱桃的NAC蛋白同源性最高。荧光定量PCR结果表明,PpNAC72在桃树各器官中均有不同程度的表达,果实中表达量最高;在不同时期的表达量也有明显变化,在桃果实生长中后期表达量最高。PpNAC72作为转录因子可能参与桃的成熟。

关键词: 桃, PpNAC72, 成熟, 表达分析

Abstract: The aims are to clone the PpNAC72 from the peach sepecies “Jinliuzaohong”,to analyze the expression profile in different development stages,and to explore its role in the peach growth. RT-PCR was used to clone the PpNAC72,and subsequently to quantitatively analyze its expression profile and bioinformatics. The results showed that the PpNAC72（GenBank accession number：MH627940）had an open reading frame sequence of 1 050 bp,encoding 349 amino acids,an isoelectric point of 8.34,and a NAM domain with typical NAC family structure. Amino acid homology analysis demonstrated that PpNAC72 had high similarity to the reported NAC of other plants,and the NAC protein had the highest homology with sweet cherry. The results of qRT-PCR revealed that PpNAC72 was expressed in different degrees in peach organs,and the expression level was the highest in fruits. The expression level was also significantly varied in different periods,and the expression level was the highest in the middle and late growth stages of peach fruit. The PpNAC72 might be involved in the maturation of peach as a transcription factor.

Key words: peach, PpNAC72, mature, expression analysis

武肖琦, 冯涛, 刘艳军, 焦思鹏, 孙攀, 杨静慧. 桃PpNAC72的克隆及表达分析[J]. 生物技术通报, 2019, 35(6): 16-23.

WU Xiao-qi, FENG Tao, LIU Yan-jun, JIAO Si-peng, SUN Pan, YANG Jing-hui. Cloning and Expression Analysis of the PpNAC72 in Peach[J]. Biotechnology Bulletin, 2019, 35(6): 16-23.

参考文献

[1] 汪祖华, 庄恩及. 中国果树志—桃卷[M]. 北京:中国林业出版社, 2001.
[2] Aida M, Ishida T, Fukaki H, et al.Genes involved in organ separation in Arabidopsis:an analysis of the cup-shaped cotyledon mutant[J]. The Plant Cell, 1997, 9(6):841-857.
[3] Pérezrodríguez P, Corrêa LGG, Rensing SA, et al.PlnTFDB:updated content and new features of the plant transcription factor database[J]. Nucleic Acids Research, 2010, 38:822-827.
[4] Pinheiro GL, Marques CS, Costa MDBL, et al.Complete inventory of soybean NAC transcription factors:sequence conservation and expression analysis uncover their distinct roles in stress response[J]. Gene, 2009, 444(1):10-23.
[5] Olsen AN, Ernst HA, Lehhio LL, et al.NAC transcription factors:structurally distinct, functionally diverse[J]. Trends in Plant Science, 2005, 10(2):79-87.
[6] Ko JH, Yang SH, Park AH, et al.ANAC012, a member of the plant-specific NAC transcription factor family, negatively regulates xylary fiber development in Arabidopsis thaliana[J]. The Plant Journal, 2007, 50(6):1035-1048.
[7] Xie Q, Frugis G, Colgan D, et al.Arabidopsis NAC1 transduces auxin signal downstream of TIR1 to promote lateral root development[J]. Genes & Development, 2000, 14(23):3024-3036.
[8] He XJ, Mu RL, Cao WH, et al.AtNAC2, a transcription factor downstream of ethylene and auxin signaling pathways, is involved in salt stress response and lateral root development[J]. The Plant Journal, 2010, 44(6):903-916.
[9] Mao C, Ding W, Wu Y, et al.Overexpression of a NAC-domain protein promotes shoot branching in rice[J]. New Phytologist, 2007, 176(2):288-298.
[10] Uauy C, Distelfeld A, Fahima T, et al.A NAC gene regulating senescence improves grain protein, zinc, and iron content in wheat[J]. Science, 2006, 314(5803):1298-1301.
[11] Guo Y, Gan S.AtNAP, a NAC family transcription factor, has an important role in leaf senescence[J]. The Plant Journal, 2010, 46(4):601-612.
[12] Jensen MK, Hagedorn PH, De TM, et al.Transcriptional regulation by an NAC(NAM-ATAF1, 2-CUC2)transcription factor attenuates ABA signalling for efficient basal defence towards Blumeria graminis f. sp. hordei in Arabidopsis[J]. The Plant Journal, 2010, 56(6):867-880.
[13] Bu Q, Jiang H, Li CB, et al.Role of the Arabidopsis thaliana NAC transcription factors ANAC019 and ANAC055 in regulating jasmonic acid-signaled defense responses[J]. Plant Signaling & Behavior, 2009, 18(5):756-767.
[14] Kim SG, Lee AK, Yoon HK, et al.A membrane-bound NAC transcription factor NTL8 regulates gibberellic acid-mediated salt signaling in Arabidopsis seed germination[J]. The Plant Journal, 2008, 55(1):77-88.
[15] Bailey CH, Hough LF.A hypothesis for the inheritance of season of ripening in progenies from certain early ripening peach varieties and selections[J]. Proc Amer Soc Hort Sci, 1959, 73:125-133.
[16] 景士西, 吴禄平. 园艺学年评[M]. 北京:科学出版社, 1996.
[17] Eduardo I, Picañol R, Rojas E, et al.Mapping of a major gene for the slow ripening character in peach:co-location with the maturity date gene and development of a candidate gene-based diagnostic marker for its selection[J]. Euphytica, 2015, 205(2):627-636.
[18] Pirona R, Eduardo I, Pacheco I, et al.Fine mapping and identification of a candidate gene for a major locus controlling maturity date in peach[J]. BMC Plant Biology, 2013, 13(1):166.
[19] Tong ZG, Gao ZH, Wang F, et al.Selection of reliable reference genes for gene expression studies in peach using real-time PCR[J]. BMC Molecular Biology, 2009, 10(1):1-13.
[20] Puranik S, Sahu PP, Srivastava PS, et al.NAC proteins:regulation and role in stress tolerance[J]. Trends in Plant Science, 2012, 17(6):369-381.
[21] Duval M, Hsieh TF, Kim SY, et al.Molecular characterization of AtNAM:A member of the Arabidopsis NAC domain superfamily
[J]. Plant Molecular Biology, 2002, 50(2):237-248.
[22] Hao YJ, Wei W, Song QX, et al.Soybean NAC transcription factors promote abiotic stress tolerance and lateral root formation in transgenic plants[J]. The Plant Journal, 2011, 68(2):302-313.
[23] Wang Z, Dane F.NAC(NAM/ATAF/CUC)transcription factors in different stresses and their signaling pathway[J]. Acta Physiologiae Plantarum, 2013, 35(5):1397-1408.
[24] 邹嘉欣, 吕楠, 朱梦丽, 等. NAC家族生物信息学分析[J]. 生物技术通讯, 2016, 14(8):1948-1964.
[25] Kou X, Watkins CB, Gan SS.Arabidopsis AtNAP regulates fruit senescence[J]. Journal of Experimental Botany, 2012, 63(17):6139-6147.
[26] Liu YZ, Baig MNR, Fan R, et al.Identification and expression pattern of a novel NAM, ATAF, and CUC-like gene from Citrus sinensis Osbeck[J]. Plant Molecular Biology Reporter, 2009, 27(3):292-297.
[27] Fujita M, Fujita Y, Maruyama K, et al.A dehydration-induced NAC protein RD26 is involved in a novel ABA-dependent stress-signaling pathway[J]. The Plant Journal, 2004, 39(6):863-876.
[28] 林菁. 辣椒转录因子CaNAC72的克隆及其功能鉴定[D]. 福建:福建农林大学, 2012.

桃PpNAC72的克隆及表达分析

Cloning and Expression Analysis of the PpNAC72 in Peach

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