Identification and functional characterization of the MdHB-1 gene promoter sequence from Malus×domestica

doi:10.1016/S2095-3119(16)61548-4

摘要/Abstract

Abstract: Homeobox 1 in Malus×domestica (MdHB-1) is a transcription factor that belongs to homeodomain-leucine zipper I (HD-Zip I) protein subfamily. According to previous reports, MdHB-1 could regulate ethylene synthesis by binding with the MdACO1 promoter, but other functions of MdHB-1 are still unknown. To reveal more clues concerning the characters of the MdHB-1 gene promoter and the functions of MdHB-1, the promoter region of MdHB-1 was cloned from the Royal Gala apple genome and recombined with the β-glucuronidase (GUS) gene in this study. This research was conducted in Nicotiana tabacum and supported by Agrobacterium-mediated transient transformation and bioinformatics analysis. Deletion analysis of the MdHB-1 promoter showed that the GUS gene could be activated by serially deleted promoters, and the activity promoted by 680 nucleotides (nt) was the lowest. The region, which is 266 nt upstream of the initiation code (ATG), was effective for GUS expression. Meanwhile, the activity of the MdHB-1 promoter (-1 057 nt), which was stronger than MdHB-1 promoter (-1 057 to -266 nt) and lack the 5´-untranslated region (5´-UTR), showed that 5´-UTR may have a positive effect on gene transcription. After the sequence analysis, the cis-acting elements that respond to hormones and environmental stresses were identified in the promoter region. The MdHB-1 promoter (1 057 nt) activity in Nicotiana tabacum was positively induced by ethrel and darkness, and it was suppressed by gibberellic acid (GA), whereas abscisic acid (ABA), salicylic acid (SA), wounding, and Pseudomonas syringae pv. tomato (DC3000) treatments revealed a slight auxo-action. These results reveal that the MdHB-1 promoter receive internal or external signals, and MdHB-1 may refer to many biological activities in apple, such as its stress response, development, and ripening.

Key words: transcription factor , HD-Zip , MdHB-1 , promoter , hormone , stress

. [J]. Journal of Integrative Agriculture, 2017, 16(08): 1730-1741.

WANG Hao-jie, JIANG Yong-hua, QI Ying-wei, DAI Jie-yu, LIU Yan-li, ZHU Xian-bo, LIU Cui-hua, Lü Yan-rong, REN Xiao-lin . Identification and functional characterization of the MdHB-1 gene promoter sequence from Malus×domestica[J]. Journal of Integrative Agriculture, 2017, 16(08): 1730-1741.

参考文献

Agalou A, Purwantomo S, Övernas E, Johannesson H, Zhu X, Estiati A, De Kam R, Engstrom P, Slamet-Ledin I, Zhu Z. 2008. A genome-wide survey of HD-Zip genes in rice and analysis of drought-responsive family members. Plant Molecular Biology, 66, 87–103.

Agarwal P, Garg V, Gautam T, Pilai B, Kanoria S, Burma P. 2014. A study on the influence of different promoter and 5´UTR (URM) cassettes from Arabidopsis thaliana on the expression level of the reporter gene beta glucuronidase in tobacco and cotton. Transgenic Research, 23, 351–363.

Aoyama T, Dong C H, Wu Y, Carabelli M, Sessa G, Ruberti I, Morelli G, Chua N. 1995. Ectopic expression of the Arabidopsis transcriptional activator Athb-1 alters leaf cell fate in tobacco. The Plant Cell, 7, 1773–1785.

Arguello-Astorga G R, Herrera-Estrella L R. 1996. Ancestral multipartite units in light-responsive plant promoters have structural features correlating with specific phototransduction pathways. Plant Physiology, 112, 1151–1166.

Ariel F, Manavella P, Dezar C, Chan R. 2007. The true story of the HD-Zip family. Trends in Plant Science, 12, 419–426.

Baker S, Wilhelm K, Thomashow M. 1994. The 5´-region of Arabidopsis thaliana cor15a has cis-acting elements that confer cold-, drought- and ABA-regulated gene expression. Plant Molecular Biology, 24, 701–713.

Ciarbelli A, Ciolfi A, Salvucci S, Ruzza V, Possenti M, Carabelli M, Fruscalzo A, Sessa G, Morelli G, Ruberti I. 2008. The Arabidopsis homeodomain-leucine zipper II gene family: Diversity and redundancy. Plant Molecular Biology, 68, 465–478.

Daraselia N D, Tarchevskaya S, Narita J O. 1996. The promoter for tomato 3-hydroxy-3-methylglutaryl coenzyme A reductase gene 2 has unusual regulatory elements that direct high-level expression. Plant Physiology, 112, 727–733.

Depege-Fargeix N, Javelle M, Chambrier P, Frangne N, Gerentes D, Perez P, Rogowsky P M, Vernoud V. 2011. Functional characterization of the HD-Zip IV transcription factor OCL1 from maize. Journal of Experimental Botany, 62, 293–305.

Dezar C A, Fedrigo G V, Chan R L. 2005. The promoter of the sunflower HD-Zip protein gene Hahb4 directs tissue-specific expression and is inducible by water stress, high salt concentrations and ABA. Plant Science, 169, 447–456.

Dezar C A, Giacomelli J, Manavella P, Ré D, Alves-Ferreira M, Baldwin I, Bonaventure G, Chan R. 2011. HAHB10, a sunflower HD-Zip II transcription factor, participates in the induction of flowering and in the control of phytohormone-mediated responses to biotic stress. Journal of Experimental Botany, 62, 1061–1076.

Diaz-De-Leon F, Klotz K, Lagrimini L. 1993. Nucleotide sequence of the tobacco (Nicotiana tabacum) anionic peroxidase gene. Plant Physiology, 101, 1117-1118.

Donald R, Cashmore A. Mutation of either G Box or I box sequences profoundly affects expression from the Arabidopsis rbcS-1A promoter. The EMBO Journal, 1990, 9, 1717-1726.

Doyle J J. 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin, 19, 11–15.

Foster E, Hattori J, Labbé H, Ouellet T, Fobert P R, James L E, Iyer V N, Miki B L. 1999. A tobacco cryptic constitutive promoter, tCUP, revealed by T-DNA tagging. Plant Molecular Biology, 41, 45–55.

Fu C H, Wang J P, Zhang C, Ma F W, Zhang J K. 2014. Cloning and function analysis of part of PGIP gene family member promoters from apple. Acta Horticulturae Sinica, 41, 2169–2178. (in Chinese)

Gehring W J, Qian Y Q, Billeter M, Furukubo-Tokunaga K, Schier A F, Resendez-Perez D, Affolter M, Otting G, Wüthrich K. 1994. Homeodomain-DNA recognition. Cell, 78, 211–23.

Green P J, Yong M H, Cuozzo M, Kano-Murakami Y, Silverstein P, Chua N H. 1988. Binding site requirements for pea nuclear protein factor GT-1 correlate with sequences required for light-dependent transcriptional activation of the rbcS-3A gene. The EMBO Journal, 7, 4035–4044.

Harrison M A. 2012. Cross-talk between phytohormone signaling pathways under both optimal and stressful environmental conditions. In: Khan N A, Nazar R, lqbal N, Anjum N. eds., Phytohormones and Abiotic Stress Tolerance in Plants. Springer, Berlin. pp. 49–76.

Higo K, Ugawa Y, Iwamoto M, Higo H. 1998. PLACE: A database of plant cis-acting regulatory DNA elements. Nucleic Acids Research, 26, 358–359.

Hiravama T, Shinozaki K. 2010. Research on plant abiotic stress responses in the post-genome era: Past, present and future. The Plant Journal, 61, 1041–1052.

Hooley R. 1994. Gibberellins: Perception, transduction and responses. Plant Molecular Biology, 26, 1529–1555.

Jefferson R A, Kavanagh T A, Bevan M W. 1987. GUS fusions: Beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. The EMBO Journal, 6, 3901-3907.

Kamo K, Kim A Y, Park S H, Joung Y H. 2012. The 5´UTR-intron of the Gladiolus polyubiquitin promoter GUBQ1 enhances translation efficiency in Gladiolus and Arabidopsis. BMC Plant Biology, 12, 79.

Kanoria S, Burma P K. 2012. A 28 nt long synthetic

5´UTR (synJ) as an enhancer of transgene expression in dicotyledonous plants. BMC Biotechnology, 12, 85.

Le H H, Nott A, Moore M J. 2003. How introns influence and enhance eukaryotic gene expression. Trends in Biochemical Sciences, 28, 215–220.

Lescot M, Déhais P, Thijs G, Marchal K, Moreau Y, Van De Peer Y, Rouzé P, Rombauts S. 2002. PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences. Nucleic Acids Research, 30, 325–327.

Li F, Lei H J, Zhao X J, Shen X J, Liu A L, Li T H. 2012. Isolation and characterization of two sorbitol transporter gene promoters in micropropagated apple plants (Malus×domestica) regulated by drought stress. Plant Growth Regulation, 68, 475–482.

Li X L. 2014. Mechanisms of the signal transduction for ethylene production in apple (Malus×domestica) fruit development and ripening. Ph D thesis. China Agricultural University, Beijing. (in Chinese)

Lin Z F, Hong Y G, Yin M A, Li C Y, Zhang K, Grierson D. 2008. A tomato HD-Zip homeobox protein, LeHB-1, plays an important role in floral organogenesis and ripening. The Plant Journal, 55, 301–310.

Liu Y, Lou Q, Xu W R, Xin Y, Bassett C, Wang Y. 2011. Characterization of a chalcone synthase (CHS) flower-specific promoter from Lilium orential ‘Sorbonne’. Plant Cell Reports, 30, 2187–2194.

Liu Y L, Jiang Y H, Wang H J, Qi Y W, Liu C H, Ren X L. 2015. Self-activation and interaction protein analysis of MdHB-1 transcription factor in apple. Plant Physiology Journal, 51, 1895–1900. (in Chinese)

Manavella P A, Arce A L, Dezar C A, Bitton F, Renou J P, Crespi M, Chan R L. 2006. Cross-talk between ethylene and drought signalling pathways is mediated by the sunflower Hahb-4 transcription factor. The Plant Journal, 48, 125–137.

Manavella P A, Dezar C A, Ariel F D, Drincovich M F, Chan R L. 2008. The sunflower HD-Zip transcription factor HAHB4 is up-regulated in darkness, reducing the transcription of photosynthesis-related genes. Journal of Experimental Botany, 59, 3143–3155.

Mauch-Mani B, Mauch F. 2005. The role of abscisic acid in plant-pathogen interactions. Current Opinion Plant Biology, 8, 409–414.

Maxson J M, Woodson W R. 1996. Cloning of a DNA-binding protein that interacts with the ethylene-responsive enhancer element of the carnation GST1 gene. Plant Molecular Biology, 31, 751–759.

Mayek-Pérez N, García-Espinosa R, López-Castaňeda C, Acosta-Gallegos J A, Simpson J. 2002. Water relations, histopathology and growth of common bean (Phaseolus vulgaris L.) during pathogenesis of Macrophomina phaseolina under drought stress. Physiological and Molecular Plant Pathology, 60, 185–195.

Morelli G, Ruberti I. 2002. Light and shade in the photocontrol of Arabidopsis growth. Trends in Plant Science, 7, 399–404.

Nash J, Luehrsen K R, Walbot V. 1990. Bronze-2 gene of maize: reconstruvtion of a wild-type allele and analysis of transcription and splicing. The Plant Cell, 2, 1039–1049.

Ohgishi M, Oka A, Morelli G, Ruberti I, Aoyama T. 2001. Negative autoregulation of the Arabidopsis homeobox gene ATHB-2. The Plant Journal, 25, 389–398.

Pastuglia M, Roby D, Dumas C, Cock J M. 1997. Rapid induction by wounding and bacterial infection of an S gene family receptor-like kinase in Brassica oleracea. The Plant Cell, 9, 1–13.

Paul S L, Jeffery L D, Michael B A, Klaus H, Wolfgang S. 1989. Inducible in vivo DNA footprints define sequences necessary for UV light activation of the parsley chalcone synthase gene. The EMBO Journal, 8, 651–656.

Qin Y F, Li D D, Wu Y J, Liu Z H, Zhang J, Zheng Y, Li X B. 2010. Three cotton homeobox genes are preferentially expressed during early seedling development and in response to phytohormone signaling. Plant Cell Reports, 29, 1147–1156.

Ré D, Dezar C, Chan R, Baldwin I, Bonaventure G. 2011. Nicotiana attenuata NaHD20 plays a role in leaf ABA accumulation during water stress, benzylacetone emission from flowers, and the timing of bolting and flower transitions. Journal of Experimental Botany, 62, 155–166.

Rodrigo L, André D, Klaus H, Wolfgang S. 1989. A phenylalanine ammonia-lyase gene from parsley: Structure, regulation and identification of elicitor and light responsive cis-acting elements. The EMBO Journal, 8, 1641–1648.

Rohde W, Dörr S, Salamini F, Becker D. 1991. Structure of a chalcone synthase gene from Hordeum vulgare. Plant Molecular Biology, 16, 1103–1106.

Roy R, Purty R, Agrawal V, Gupta S. 2006. Promoterless gus gene shows leaky β-glucuronidase activity during transformation of tomato with bspA gene for drought tolerance. Biologia Plantarum, 50, 352–358.

Ruberti I, Sessa G, Lucchetti S, Morelli G. 1991. A novel class of plant proteins containing a homeodomain with a closely linked leucine zipper motif. The EMBO Journal, 10, 1787.

Shahmuradov I A, Gammerman A J, Hancock J M, Bramley P M, Solovyev V V. 2003. PlantProm: A database of plant promoter sequences. Nucleic Acids Research, 31, 114–117.

Shan D P, Huang J G, Yang Y T, Guo Y H, Wu C A, Yang G D, Gao Z, Zheng C C. 2007. Cotton GhDREB1 increases plant tolerance to low temperature and is negatively regulated by gibberellic acid. New Phytologist, 176, 70–81.

Sparkes I A, Runions J, Kearns A. Hawes C. 2006. Rapid, Transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants. Nature Protocols, 1, 2019–2025.

Springer P S. 2000. Gene traps: Tools for plant development and genomics. The Plant Cell, 12, 1007–1020.

Urao T, Yamaguchi-Shinozaki K, Urao S, Shinozaki K. 1993. An Arabidopsis MYB homolog is induced by dehydration stress and its gene product binds to the conserved MYB recogniton sequence. The Plant Cell, 5, 1529–1539.

Valdés A E, Övernas E, Johanssoon H, Rada-Iglesias A, Engstrom P. 2012. The homeodomain-leucine zipper (HD-Zip) class I transcription factors ATHB7 and ATHB12 modulate abscisic acid signalling by regulating protein phosphatase 2C and abscisic acid receptor gene activities. Plant Molecular Biology, 80, 405–418.

Viana A A, Fragoso R R, Guimaraes L M, Pontes N, Oliveira-Neto O B, Artico S, Nardeli S M, Alves-Ferreira M, Batista J A, Silva M C. 2011. Isolation and functional characterization of A cotton ubiquitination-related promoter and 5´UTR that drives high levels of expression in root and flower tissues. BMC Biotechnology, 11, 115.

Vidhasekaran P. 2015. Salicylic acid signaling in plant innate immunity. In: Plant Hormone Signaling Systems in Plant Innate Immunity. Springer, Netherlands. pp. 27–122.

Walker J C, Howard E A, Dennis E S, Peacock W J. 1987. DNA sequences required for anaerobic expression of maize alcohol dehydrogenase 1 gene. Proceedings of the National Academy of Sciences of the United States of America, 84, 6624–6628.

Wang H J, Jiang Y H, Liu Y L, Qi Y W, Liu C H, Ren X L. 2016. Response analysis of the ethylene-responsive element of MdHB-1 promoter to exogenous ethephon. Plant Physiology Journal, 52, 365–371. (in Chinese)

Wei Q, Kuai B, Hu P, Ding Y L. 2012. Ectopic-overexpression of an HD-Zip IV transcription factor from Ammopiptanthus mongolicus (leguminosae) promoted upward leaf curvature and non-dehiscent anthers in Arabidopsis thaliana. The Plant Cell, Tissue and Organ Culture, 110, 299–306.

Wen X H, Jiang Y H, Kang J, Wang H J, Ren X L. 2015a. A genome-wide survey of HD-Zip I genes and their responses to hormone in apple. Scientia Agricultura Sinica, 48, 4314-4322. (in Chinese)

Wen X H, Jiang Y H, Zhou K, Ren X L. 2015b. Effects of virus-induced MdHB-1 silencing on apple fruit ripening. Acta Agriculturae Boreali-occidentalis Sinica, 24, 113–119. (in Chinese)

Xu W R, Yu Y H, Ding J H, Hua Z Y, Wang Y J. 2010. Characterization of a novel stilbene synthase promoter involved in pathogen- and stress-inducible expression from Chinese wild Vitis pseudoreticulata. Planta, 231, 475–487.

Yang Y N, Li R G, Qi M. 2000. In vivo analysis of plant promoters and transcription factors by agroinfiltration of tobacco leaves. The Plant Journal, 22, 543–551.

Yoshida K, Mohri T, Nishiguchi M, Tazaki K. 2002. Robinia pseudoacacia inner-bark lectin promoter expresses GUS also predominantly in phloem of transgenic tobacco. Journal of Plant Physiology, 159, 757–764.

Yu Q P, Yin M A, Ren X L, Jiang Y H. 2012. Cloning and eukaryotic expression vector construction of transcription factor MdHB-1 gene in apple. Journal of Northwest A&F University (Nature Science), 40, 154–160. (in Chinese)

Zhu M, Yin M A, Ye H, Ren X L. 2014. Expression patterns of ACO1 gene and transcription factor MdHB-1 in apple. Food Science, 35, 23. (in Chinese)