Abstract
The Arabidopsis gene AINTEGUMENTA (At-ANT) functions in cell proliferation and organ growth. The ANT protein has two copies of the AP2 domains, R1 and R2. Recently, a partial cDNA sequence of the At-ANT homolog in Antirrhinum majus (Am-ANT) was reported (Delgado-Benarroch et al., 2009). Here, we used virus-induced gene silencing (VIGS) to analyze the function of the reported Am-ANT. We then determined the open reading frame (ORF) of Am-ANT and its predicted amino acid sequence. We induced VIGS using the Cucumber mosaic virus vector (CMV-A1) that contained partial sequence of Am-ANT (A1:ANT) and suppressed the level of Am-ANT mRNA and noted for any phenotypic changes. The function of Am-ANT was very similar to that of At-ANT. The A1:ANT-infected Antirrhinum plants had smaller floral organs and leaves, even though cell sizes were unchanged in flowers and larger in leaves. The CMV-based VIGS showed that the isolated Am-ANT gene was indeed functional in cell proliferation and organ growth as observed for At-ANT. In conclusion, we found that the CMV vector had the advantage of systemically infecting A. majus without severe symptoms for functional analysis of A. majus genes.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Literature Cited
Baker, S.C., K. Robinson-Beers, J.M. Villanueva, J.C. Gaiser, and C.S. Gasser. 1997. Interactions among genes regulating ovule development in Arabidopsis thaliana. Genetics 145:1109–1124.
Burch-Smith, T.M., J.C. Anderson, G.B. Martin, and S.P. Dinesh-Kumar. 2004. Applications and advantages of virusinduced gene silencing for gene function studies in plants. The Plant J. 39:734–746.
Darwin, C.R. 1868. Variation of animals and plants under domestication. John Murray, London.
Delgado-Benarroch, L., B. Causier, J. Weiss, and M. Egea-Cortines. 2009. FORMOSA controls cell division and expansion during floral development in Antirrhinum majus. Planta 229:1219–1229.
Elliott, R.C., A.S. Betzner, E. Huttner, M.P. Oakes, W.Q. Tucker, D. Gerentes, P. Perez, and D.R. Smyth. 1996. AINTEGUMENTA, an APETALA2-like gene of Arabidopsis with pleiotropic roles in ovule development and floral organ growth. Plant Cell 8:155–168.
Goto, K., A. Kanazawa, M. Kusaba, and C. Masuta. 2003. A simple and rapid method to detect plant siRNAs using nonradioactive probes. Plant Mol. Biol. Rep. 21:51–58.
Hamilton, A.J. and D.C. Baulcombe. 1999. A species of small antisense RNA in posttranscriptional gene silencing in plants. Science 286:950–952.
Horiguchi, G., A. Ferjani, U. Fujikura, and H. Tsukaya. 2006. Coordination of cell proliferation and cell expansion in the control of leaf size in Arabidopsis thaliana. J. Plant Res. 119:37–42.
Hudson, A., J. Critchley, and Y. Erasmus. 2008. The Genus Antirrhinum (Snapdragon): A flowering plant model for evolution and development. Cold Spring Harb. Protoc. doi 10.1101/pdb.emo100.
Kim, B., C. Masuta, H. Matsuura, H. Takahashi, and T. Inukai. 2008. Veinal necrosis induced by Turnip mosaic virus infection in Arabidopsis is a form of defense response accompanying HR-like cell death. Mol. Plant-Microbe Interact. 21:260–268.
Klucher, K.M., H. Chow, L. Reiser, and R.L. Fischer. 1996. The AINTEGUMENTA gene of Arabidopsis required for ovule and female gametophyte development is related to the floral homeotic gene APETALA2. Plant Cell 8:137–153.
Krizek, B.A. 1999. Ectopic expression of AINTEGUMENTA in Arabidopsis plants results in increased growth of floral organs. Dev. Genet. 25:224–236.
Krizek, B.A. 2003. AINTEGUMENTA utilizes a mode of DNA recognition distinct from that used by proteins containing a single AP2 domain. Nucleic Acids Res. 31:1859–1868.
Mizukami, Y. and R.L. Fischer. 2000. Plant organ size control: AINTEGUMENTA regulates growth and cell numbers during organogenesis. Proc. Natl. Acad. Sci. U.S.A. 97:942–947.
Mizukami, Y. 2001. A matter of size: developmental control of organ size in plants. Curr. Opin. Plant Biol. 4:533–539.
Nagamatsu, A., C. Masuta, M. Senda, H. Matsuura, A. Kasai, J.S. Hong, K. Kitamura, J. Abe, and A. Kanazawa. 2007. Functional analysis of soybean genes involved in flavonoid biosynthesis by virus induced gene silencing. Plant Biotechnol. 5:778–790.
Nole-Wilson, S. and B.A. Krizek. 2000. DNA binding properties of the Arabidopsis floral development protein AINTEGUMENTA. Nucleic Acids Res. 28:4076–4082.
Otagaki, S., M. Arai, A. Takahashi, K. Goto, J.S. Hong, C. Masuta, and A. Kanazawa. 2006. Rapid induction of transcriptional and post-transcriptional gene silencing using a novel Cucumber mosaic virus vector. Plant Biotechnol. 23:259–265.
Preston, J.C. and L.C. Hileman. 2010. SQUAMOSA-PROMOTER BINDING PROTEIN 1 initiates flowering in Antirrhinum majus through the activation of meristem identity genes. The Plant J. 62:704–712.
Rieu, I., M. Bots, C. Mariani, and K.A.P. Weterings. 2005. Isolation and expression analysis of a tobacco AINTEGUMENTA ortholog (NtANTL). Plant Cell Physiol. 46:803–805.
Schneitz, K., M. Hülskamp, S.D. Kopczak, and R.E. Pruitt. 1997. Dissection of sexual organ ontogenesis: a genetic analysis of ovule development in Arabidopsis thaliana. Development 124:1367–1376.
Shang, Y., K.E. Schwinn, M.J. Bennett, D.A. Hunter, T.L. Waugh, N.N. Pathirana, D.A. Brummell, P.E. Jameson, and K.M. Davies. 2007. Methods for transient assay of gene function in floral tissues. Plant Methods 3:1.
Suzuki, M., S. Kuwata, J. Kataoka, C. Masuta, N. Nitta, and Y. Takanami. 1991. Functional analysis of deletion mutants of Cucumber mosaic virus RNA3 using an in vitro transcription system. Virol. 183:106–113.
Author information
Authors and Affiliations
Corresponding author
Additional information
These authors are contributed equally to this work.
Rights and permissions
About this article
Cite this article
Kim, B.M., Inaba, Ji. & Masuta, C. Virus induced gene silencing in Antirrhinum majus using the Cucumber mosaic virus vector: Functional analysis of the AINTEGUMENTA (Am-ANT) gene of A. majus . Hortic. Environ. Biotechnol. 52, 176–182 (2011). https://doi.org/10.1007/s13580-011-0172-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13580-011-0172-y