Transcription factors (TFs) preferentially expressed in developing endosperm were identified by searching a maize transcriptome database. One thousand and two hundred TFs were annotated, 414 of which were shown to express in the seeds. Of these, 113 presented preferential expression in developing endosperm. Among the endosperm-preferred TFs 13% were represented by members of the zinc-finger family, 17% by members of the bZIP family and 11% by members of the NAC family. Among the NAC family TFs, we identified APN-1, whose expression was detected 5 days after pollination (DAP) and peaked at 20 DAP. Analysis of the promoter sequence of gene encoding APN-1 revealed the presence of cis-elements related to endosperm-specificity, abscisic acid (ABA) and gibberellic acid (GA) signalling. Transient expression analysis of a construct containing the β-glucuronidase gene driven by the Apn-1 gene promoter showed that expression was restricted to the aleurone cell layer. Analysis of mRNA levels in developing maize endosperm treated with hormones revealed that the expression of the gene encoding APN-1 was stimulated by ABA and inhibited by GA. © 2010 Association of Applied Biologists.1581115129Aida, M., Ishida, T., Fukaki, H., Fujisawa, H., Tasaka, M., Genes involved in organ separation in Arabidopsis: An analysis of the cup-shaped cotyledon mutant (1997) Plant Cell, 9, pp. 841-857Bateman, A., Coin, L., Durbin, R., Finn, R.D., Hollich, V., Griffiths-Jones, S., Khanna, A., Eddy, S.R., The Pfam protein families database (2004) Nucleic Acids Research, 32, pp. D138-D141Benson, D.A., Karsch-Mizrachi, I., Lipman, D.J., Ostell, J., Wheeler, D.L., GenBank (2005) Nucleic Acids Research, 33, pp. D34-D38Bobb, A.J., Chern, M.S., Bustos, M.M., Conserved RY-repeats mediate transactivation of seed-specific promoters by the developmental regulator PvALF (1997) Nucleic Acids Research, 25, pp. 641-647Bowman, J.L., The YABBY gene family and abaxial cell fate (2000) Current Opinion in Plant Biology, 3, pp. 17-22De Freitas, F.A., Yunes, J.A., Da Silva, M.J., Arruda, P., Leite, A., Structural characterization and promoter activity analysis of the gamma-kafirin gene from sorghum (1994) Molecular & General Genetics, 245, pp. 177-186Duval, M., Hsieh, T.F., Kim, S.Y., Thomas, T.L., Molecular characterization of AtNAM: A member of the Arabidopsis NAC domain superfamily (2002) Plant Molecular Biology, 50, pp. 237-248Ezcurra, I., Wycliffe, P., Nehlin, L., Ellerstrom, M., Rask, L., Transactivation of the Brassica napus napin promoter by ABI3 requires interaction of the conserved B2 and B3 domains of ABI3 with different cis-elements: B2 mediates activation through an ABRE, whereas B3 interacts with an RY/G-box (2000) The Plant Journal, 24, pp. 57-66Fujita, M., Fujita, Y., Maruyama, K., Seki, M., Hiratsu, K., Ohme-Takagi, M., Tran, L.-S.P., Shinozaki, K., A dehydration-induced NAC protein, RD26, is involved in a novel ABA-dependent stress-signaling pathway (2004) The Plant Journal, 39, pp. 863-876Giroux, M.J., Boyer, C., Feix, G., Hannah, L.C., Coordinated transcriptional regulation of storage product genes in the maize endosperm (1994) Plant Physiology, 106, pp. 713-722Gomez-Cadenas, A., Zentella, R., Sutliff, T.D., Ho, T.D.H., Involvement of multiple cis-elements in the regulation of GA responsive promoters: Definition of a new cis-element in the Amy32b gene promoter of barley (Hordeum vulgare) (2001) Physiologia Plantarum, 112, pp. 211-216Gong, W., Shen, Y.P., Ma, L.G., Pan, Y., Du, Y.L., Wang, D.H., Yang, J.Y., Zhu, Y.X., Genome-wide ORFeome cloning and analysis of Arabidopsis transcription factor genes (2004) Plant Physiology, 135, pp. 773-782Greve, K., La Cour, T., Jensen, M.K., Poulsen, F.M., Skriver, K., Interactions between plant RING-H2 and plant-specific NAC (NAM/ATAF1/2/CUC2) proteins: RING-H2 molecular specificity and cellular localization (2003) Biochemical Journal, 371, pp. 97-108Gubler, F., Kalla, R., Roberts, J.K., Jacobsen, J.V., Gibberellin-regulated expression of a myb gene in barley aleurone cells - Evidence for Myb transactivation of a high-pl alpha-amylase gene promoter (1995) Plant Cell, 7, pp. 1879-1891Guo, M., Rupe, M.A., Danilevskaya, O.N., Yang, X.F., Hut, Z.H., Genome-wide mRNA profiling reveals heterochronic allelic variation and a new imprinted gene in hybrid maize endosperm (2003) The Plant Journal, 36, pp. 30-44Higo, K., Ugawa, Y., Iwamoto, M., Korenaga, T., Plant cis-acting regulatory DNA elements (PLACE) database (1999) Nucleic Acids Research, 27, pp. 297-300Hobo, T., Asada, M., Kowyama, Y., Hattori, T., ACGT-containing abscisic acid response element (ABRE) and coupling element 3 (CE3) are functionally equivalent (1999) The Plant Journal, 19, pp. 679-689Hoecker, U., Vasil, I.K., McCarty, D.R., Signaling from the embryo conditions Vp1-mediated repression of alpha-amylase genes in the aleurone of developing maize seeds (1999) The Plant Journal, 19, pp. 371-377Jefferson, R.A., Kavanagh, T.A., Bevan, M.W., Gus fusions - Beta-glucuronidase as a sensitive and versatile gene fusion marker in higher-plants (1987) Embo Journal, 6, pp. 3901-3907Jofuku, K.D., Omidyar, P.K., Gee, Z., Okamuro, J.K., Control of seed mass and seed yield by the floral homeotic gene APETALA2 (2005) Proceedings of the National Academy of Sciences of the United States of America, 102, pp. 3117-3122Kiyosue, T., Ohad, N., Yadegari, R., Hannon, M., Dinneny, J., Wells, D., Katz, A., Fischer, R.L., Control of fertilization-independent endosperm development by the MEDEA polycomb gene in Arabidopsis (1999) Proceedings of the National Academy of Sciences of the United States of America, 96, pp. 4186-4191Kohler, C., Grossniklaus, U., Epigenetic inheritance of expression states in plant development: The role of Polycomb group proteins (2002) Current Opinion in Cell Biology, 14, pp. 773-779Kohler, C., Hennig, L., Bouveret, R., Gheyselinck, J., Grossniklaus, U., Gruissem, W., Arabidopsis MSI1 is a component of the MEA/FIE Polycomb group complex and required for seed development (2003) Embo Journal, 22, pp. 4804-4814Kosugi, S., Ohashi, Y., Nakajima, K., Arai, Y., An improved assay for β-glucuronidase in transformed cells - Methanol almost completely suppresses a putative endogenous β-glucuronidase activity (1990) Plant Science, 70, pp. 133-140Lescot, M., Dehais, P., Thijs, G., Marchal, K., Moreau, Y., Van De Peer, Y., Rouze, P., Rombauts, S., PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences (2002) Nucleic Acids Research, 30, pp. 325-327Livak, K.J., Schmittgen, T.D., Analysis of relative gene expression data using real-time quantitative PCR and the 2-DDCt method (2001) Methods, 25, pp. 402-408Lopes, M.A., Larkins, B.A., Endosperm origin, development, and function (1993) Plant Cell, 5, pp. 1383-1399Manning, K., Isolation of nucleic acids from plants by differential solvent precipitation (1991) Analytical Biochemistry, 195, pp. 45-50Matys, V., Fricke, E., Geffers, R., Gossling, E., Haubrock, M., Hehl, R., Hornischer, K., Wingender, E., TRANSFAC (R): Transcriptional regulation, from patterns to profiles (2003) Nucleic Acids Research, 31, pp. 374-378Neill, S.J., Horgan, R., Parry, A.D., The carotenoid and abscisic acid content of viviparous kernels and seedlings of Zea mays L (1986) Planta, 169, pp. 87-96Ober, E.S., Setter, T.L., Water deficit induces abscisic acid accumulation in endosperm of maize viviparous mutants (1992) Plant Physiology, 98, pp. 353-356Ohto, M., Fischer, R.L., Goldberg, R.B., Nakamura, K., Harada, J.J., Control of seed mass by APETALA2 (2005) Proceedings of the National Academy of Sciences of the United States of America, 102, pp. 3123-3128Olsen, O.A., Endosperm development: Cellularization and cell fate specification (2001) Annual Review of Plant Physiology and Plant Molecular Biology, 52, pp. 233-267Pabo, C.O., Sauer, R.T., Transcription factors - Structural families and principles of DNA recognition (1992) Annual Review of Biochemistry, 61, pp. 1053-1095Price, J., Laxmi, A., St Martin, S.K., Jang, J.C., Global transcription profiling reveals multiple sugar signal transduction mechanisms in Arabidopsis (2004) Plant Cell, 16, pp. 2128-2150Riechmann, J.L., Heard, J., Martin, G., Reuber, L., Jiang, C.-Z., Keddie, J., Adam, L., Yu, G.-L., Arabidopsis transcription factors: Genome-wide comparative analysis among eukaryotes (2000) Science, 290, pp. 2105-2110Riechmann, J.L., Ratcliffe, O.J., A genomic perspective on plant transcription factors (2000) Current Opinion in Plant Biology, 3, pp. 423-434Robertson, M., Two transcription factors are negative regulators of gibberellin response in the HvSPY-signaling pathway in barley aleurone (2004) Plant Physiology, 136, pp. 2747-2761Schwechheimer, C., Bevan, M., The regulation of transcription factor activity in plants (1998) Trends in Plant Science, 3, pp. 378-383Souer, E., Vanhouwelingen, A., Kloos, D., Mol, J., Koes, R., The no apical meristem gene of petunia is required for pattern formation in embryos and flowers and is expressed at meristem and primordia boundaries (1996) Cell, 85, pp. 159-170Sung, Z.R., Chen, L.J., Moon, Y.H., Lertpiriyapong, K., Mechanisms of floral repression in Arabidopsis (2003) Current Opinion in Plant Biology, 6, pp. 29-35Suzuki, M., Latshaw, S., Sato, Y., Settles, A.M., Koch, K.E., Hannah, L.C., Kojima, M., McCarty, D.R., The maize Viviparous8 locus, encoding a putative ALTERED MERISTEM PROGRAM1-like peptidase, regulates abscisic acid accumulation and coordinates embryo and endosperm development (2008) Plant Physiology, 146, pp. 1193-1206Verza, N.C., Silva, T.R., Neto, G.C., Nogueira, F.T.S., Fisch, P.H., De Rosa, V.E., Rebello, M.M., Arruda, P., Endosperm-preferred expression of maize genes as revealed by transcriptome-wide analysis of expressed sequence tags (2005) Plant Molecular Biology, 59, pp. 363-374White, C.N., Proebsting, W.M., Hedden, P., Rivin, C.J., Gibberellins and seed development in maize. I. Evidence that gibberellin/abscisic acid balance governs germination versus maturation pathways (2000) Plant Physiology, 122, pp. 1081-1088Yanagisawa, S., Akiyama, A., Kisaka, H., Uchimiya, H., Miwa, T., Metabolic engineering with Dof1 transcription factor in plants, improved nitrogen assimilation and growth under low-nitrogen conditions (2004) Proceedings of the National Academy of Sciences of the United States of America, 101, pp. 7833-7838Yunes, J.A., Neto, G.C., Da Silva, M.J., Leite, A., Ottoboni, L.M.M., Arruda, P., The transcriptional activator Opaque2 recognizes 2 different target sequences in the 22-Kd-like alpha-prolamin genes (1994) Plant Cell, 6, pp. 237-249Yunes, J.A., Vettore, A.L., Da Silva, M.J., Leite, A., Arruda, P., Cooperative DNA binding and sequence discrimination by the Opaque2 bZIP factor (1998) Plant Cell, 10, pp. 1941-1955Zimmermann, R., Werr, W., Pattern formation in the monocot embryo as revealed by NAM and CUC3 orthologues from Zea mays L (2005) Plant Molecular Biology, 58, pp. 669-685