Abstract
The Antirrhinum DEFH125 MADS-box protein is expressed in maturing pollen and thus likely participates in the regulation of pollen development. Here, we describe the characterization of a 2.5 kbp promoter fragment conferring pollen-specific GUS expression in Antirrhinum, as well as in the distantly related species Arabidopsis. Taking advantage of the higher sensitivity of the diphtheria toxin A-chain (DTA) reporter gene assay, onset of DEFH125 promoter activity could be defined to start at the late unicellular microspore stage. Stamen development in Antirrhinum is governed by the class B MADS-box genes DEFICIENS (DEF) and GLOBOSA (GLO). The respective proteins form a heterodimer and are expressed throughout stamens, except for microspores. Complementary expression patterns of DEFH125 and DEF/GLO during later stamen development tempted us to investigate whether the DEF/GLO heterodimer might bind the DEFH125 promoter and could thus be involved in repressing the DEFH125 expression. The ChIP technique was applied to investigate protein/DNA interactions occurring in vivo. We report the identification of a 200 bp DEFH125 promoter fragment that is in vivo bound by DEF and GLO proteins. This fragment contains a CArG-box motif, known to mediate DNA binding of MADS-box proteins. Implications for a likely function of DEF and GLO in the transcriptional control of DEFH125 are discussed.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- ChIP:
-
Chromatin immunoprecipitation
- DEF :
-
DEFICIENS
- DEFH125 :
-
DEFICIENS HOMOLOG 125
- DTA:
-
Diphtheria toxin A-chain
- GLO :
-
GLOBOSA
- GUS:
-
β-Glucuronidase
References
Alvarez-Buylla ER, Liljegren SJ, Pelaz S, Gold SE, Burgeff C, Ditta GS, Vergara-Silva F, Yanofsky MF (2002) MADS-box gene evolution beyond flowers: expression in pollen, endosperm, guard cells, roots and trichomes. Plant J 24:457–466
Bate N, Twell D (1998) Functional architecture of a late pollen promoter: pollen-specific transcription is developmentally regulated by multiple stage-specific and co-dependent activator elements. Plant Mol Biol 37:859–869
Becker D, Kemper E, Schell J, Masterson R (1992) New plant binary vectors with selectable markers located proximal to the left T-DNA border. Plant Mol Biol 20:1195–1197
Bey M, Stüber K, Fellenberg K, Schwarz-Sommer Z, Sommer H, Saedler H, Zachgo S (2004) Characterization of Antirrhinum petal development and identification of target genes of the class B MADS box gene DEFICIENS. Plant Cell 16:3197–3215
Chen Y-CS, McCormick S (1996) Sidecar pollen, an Arabidopsis thaliana male gametophytic mutant with aberrant cell divisions during pollen development. Development 122:3243–3253
Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743
Coen ES, Meyerowitz EM (1991) The war of the whorls: genetic interactions controlling flower development. Nature 353:31–37
Day CD, Galgoci BF, Irish VF (1995) Genetic ablation of petal and stamen primordia to elucidate cell interactions during floral development. Development 121:2887–2895
Egea-Cortines M, Saedler H, Sommer H (1999) Ternary complex formation between the MADS-box proteins SQUAMOSA, DEFICIENS and GLOBOSA is involved in the control of floral architecture in Antirrhinum majus. EMBO J 18:5370–5379
Gomez-Mena C, de Folter S, Costa MM, Angenent GC, Sablowski R (2005) Transcriptional program controlled by the floral homeotic gene AGAMOUS during early organogenesis. Development 132:429–438
Heidmann I, Efremova N, Saedler H, Schwarz-Sommer Z (1998) A protocol for transformation and regeneration of Antirrhinum majus. Plant J 13:723–728
Hill TA, Day CD, Zondlo SC, Thackeray AG, Irish VF (1998) Discrete spatial and temporal cis-acting elements regulate transcription of the Arabidopsis floral homeotic gene APETALA3. Development 125:1711–1721
Honma T, Goto K (2001) Complexes of MADS-box proteins are sufficient to convert leaves into floral organs. Nature 409:525–529
Honys D, Twell D (2003) Comparative analysis of the Arabidopsis pollen transcriptome. Plant Physiol 132:640–652
Honys D, Twell D (2004) Transcriptome analysis of haploid male gametophyte development in Arabidopsis. Genome Biol 5:R85
Jack T, Fox GL, Meyerowitz EM (1994) Arabidopsis homeotic gene APETALA3 ectopic expression: transcriptional and post-transcriptional regulation determine floral organ identity. Cell 76:703–716
McCormick S (2004) Control of male gametophyte development. Plant Cell 16(Suppl):S142–S153
Miller R (1986) Beyond ANOVA, basics of applied statistics. Wiley, New York
Müller BM (2003) Doktorarbeit (PhD thesis), University of Cologne
Orlando V, Strutt H, Paro R (1997) Analysis of chromatin structure by in vivo formaldehyde cross-linking. Methods 11:205–214
Pina C, Pinto F, Feijó JA, Becker JD (2005) Gene family analysis of the Arabidopsis pollen transcriptome reveals biological implications for cell growth, division control, and gene expression regulation. Plant Physiol 138:744–756
Sanders PM, Bui AQ, Weterings K, McIntire KN, Hsu Y-C, Lee PY, Truong MT, Beals TP, Goldberg RB (1999) Anther developmental defects in Arabidopsis thaliana male-sterile mutants. Sex Plant Reprod 11:297–322
Schiefthaler U, Balasubramanian S, Sieber P, Chevalier D, Wisman E, Schneitz K (1999) Molecular analysis of NOZZLE, a gene involved in pattern formation and early sporogenesis during sex organ development in Arabidopsis thaliana. Proc Natl Acad Sci USA 96:11664–11669
Schreiber DN, Bantin J, Dresselhaus T (2004) The MADS box transcription factor ZmMADS2 is required for anther and pollen maturation in maize and accumulates in apoptotic bodies during anther dehiscence. Plant Physiol 134:1069–1079
Schwartz YB, Kahn TG, Pirrotta V (2005) Characteristic low density and shear sensitivity of cross-linked chromatin containing polycomb complexes. Mol Cell Biol 25:432–439
Schwarz-Sommer Z, Huijser P, Nacken W, Saedler H, Sommer H (1990) Genetic control of flower development by homeotic genes in Antirrhinum majus. Science 250:931–939
Schwarz-Sommer Z, Hue I, Huijser P, Flor P, Hansen R, Tetens F, Lönnig WE, Saedler H, Sommer H (1992) Characterization of the Antirrhinum floral homeotic MADS-box gene DEFICIENS: evidence for DNA binding and autoregulation of its persistent expression throughout flower development. EMBO J 11:251–263
Scott RJ, Spielman M, Dickinson HG (2004) Stamen structure and function. Plant Cell 16(Suppl):S46–S60
Sommer H, Beltrán JP, Huijser P, Pape H, Lönnig WE, Saedler H, Schwarz-Sommer Z (1990) DEFICIENS, a homeotic gene involved in the control of flower morphogenesis in Antirrhinum majus: the protein shows homology to transcription factors. EMBO J 9:605–613
Sorensen AM, Krober S, Unte US, Huijser P, Dekker K, Saedler H (2003) The Arabidopsis ABORTED MICROSPORES (AMS) gene encodes a MYC class transcription factor. Plant J 33:413–423
Theißen G, Saedler H (2001) Floral quartets. Nature 409:469–471
Tilly JJ, Allen DW, Jack T (1998) The CArG boxes in the promoter of the Arabidopsis floral organ identity gene APETALA3 mediate diverse regulatory effects. Development 125:1647–1657
Tröbner W, Ramirez L, Motte P, Hue I, Huijser P, Lönnig WE, Saedler H, Sommer H, Schwarz-Sommer Z (1992) GLOBOSA: a homeotic gene which interacts with DEFICIENS in the control of Antirrhinum floral organogenesis. EMBO J 11:4693–4704
Turck F, Zhou A, Somssich IE (2004) Stimulus-dependent, promoter-specific binding of transcription factor WRKY1 to its native promoter and the defense-related gene PcPR1-1 in parsley. Plant Cell 16:2573–2585
Twell D (1995) Diphtheria toxin-mediated cell ablation in developing pollen: vegetative cell ablation blocks generative cell migration. Protoplasma 187:144–154
William DA, Su Y, Smith MR, Lu M, Baldwin DA, Wagner D (2004) Genomic identification of direct target genes of LEAFY. Proc Natl Acad Sci USA 101:1775–1780
Wilson ZA, Morroll SM, Dawson J, Swarup R, Tighe PJ (2001) The Arabidopsis MALE STERILITY1 (MS1) gene is a transcriptional regulator of male gametogenesis, with homology to the PHD-finger family of transcription factors. Plant J 28:27–39
Yamaizumi M, Mekada E, Uchida T, Okada Y (1978) One molecule of diphtheria toxin fragment A introduced into a cell can kill the cell. Cell 15:245–250
Yamamoto Y, Nishimura M, Hara-Nishimura I, Noguchi T (2003) Behavior of vacuoles during microspore and pollen development in Arabidopsis thaliana. Plant Cell Physiol 44:1192–1201
Yang WC, Ye D, Xu J, Sundaresan V (1999) The SPOROCYTELESS gene of Arabidopsis is required for initiation of sporogenesis and encodes a novel nuclear protein. Genes Dev 13:2108–2117
Zachgo S, de Andrade Silva E, Motte P, Tröbner W, Saedler H, Schwarz-Sommer Z (1995) Functional analysis of the Antirrhinum floral homeotic DEFICIENS gene in vivo and in vitro by using a temperature-sensitive mutant. Development 121:2861–2875
Zachgo S, Saedler H, Schwarz-Sommer Z (1997) Pollen-specific expression of DEFH125, a MADS-box transcription factor in Antirrhinum with unusual features. Plant J 11:1043–1050
Zachgo S, Perbal MC, Saedler H, Schwarz-Sommer Z (2000) In situ analysis of RNA and protein expression in whole mounts facilitates detection of floral gene expression dynamics. Plant J 23:697–702
Acknowledgements
The authors thank Dr. Hans Sommer, MPIZ, for support with the molecular work. A.L. received a scholarship from the DFG (Graduierten Kolleg, ‘Molecular analysis of developmental processes’). This work was supported by a grant from the DFG to S.Z. (ZA 259/3–1).
Author information
Authors and Affiliations
Corresponding author
Additional information
Andrea Lauri and Shuping Xing contributed equally to the work.
Rights and permissions
About this article
Cite this article
Lauri, A., Xing, S., Heidmann, I. et al. The pollen-specific DEFH125 promoter from Antirrhinum is bound in vivo by the MADS-box proteins DEFICIENS and GLOBOSA. Planta 224, 61–71 (2006). https://doi.org/10.1007/s00425-005-0193-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00425-005-0193-9