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Floral induction in tissue culture: a system for the analysis of LEAFY-dependent gene regulation

Wagner, Doris and Wellmer, Frank and Dilks, Kieran and William, Dilusha and Smith, Michael R. and Kumar, Prakash P. and Riechmann, José Luis and Greenland, Andrew J. and Meyerowitz, Elliot M. (2004) Floral induction in tissue culture: a system for the analysis of LEAFY-dependent gene regulation. Plant Journal, 39 (2). pp. 273-282. ISSN 0960-7412. doi:10.1111/j.1365-313X.2004.02127.x.

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We have developed a versatile floral induction system that is based on ectopic overexpression of the transcription factor LEAFY (LFY) in callus. During shoot regeneration, flowers or floral organs are formed directly from root explants without prior formation of rosette leaves. Morphological and reporter gene analyses show that leaf-like structures are converted to floral organs in response to LFY activity. Thus, increased levels of LFY activity are sufficient to bypass normal vegetative development and to direct formation of flowers in tissue culture. We found that about half of the cultured cells respond to inducible LFY activity with a rapid upregulation of the known direct target gene of LFY, APETALA1 (AP1). This dramatic increase in the number of LFY-responsive cells compared to whole plants suggested that the tissue culture system could greatly facilitate the analysis of LFY-dependent gene regulation by genomic approaches. To test this, we monitored the gene expression changes that occur in tissue culture after activation of LFY using a flower-specific cDNA microarray. Induction of known LFY target genes was readily detected in these experiments. In addition, several other genes were identified that had not been implicated in signaling downstream of LFY before. Thus, the floral induction system is suitable for the detection of low abundance transcripts whose expression is controlled in an LFY-dependent manner.

Item Type:Article
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Meyerowitz, Elliot M.0000-0003-4798-5153
Additional Information:© 2004 Blackwell Publishing Ltd. Received 22 February 2004; revised 20 April 2004; accepted 28 April 2004. We thank Yanhui Su and Meina Lu for technical assistance as well as Dr Annick Dubois, Dr Márcio Alves-Ferreira, Vijaya Rao, and Ali Mortazavi for invaluable help during microarray analyses. We are grateful to Chang Seob Kwon and members of the Meyerowitz laboratory for discussion and for comments on the manuscript. F.W. was supported by an Emmy Noether fellowship of the Deutsche Forschungsgemeinschaft. This research was supported by NSF grant IBN 130804 to D.W., by US Department of Energy grant FG03-88ER13873 to E.M.M. and by the Millard and Muriel Jacobs Genetics and Genomics Laboratory at the California Institute of Technology.
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Deutsche Forschungsgemeinschaft (DFG)UNSPECIFIED
Department of EnergyDE-FG03-88ER13873
Caltech Millard and Muriel Jacobs Genetics and Genomics LaboratoryUNSPECIFIED
Subject Keywords:flower development; floral induction; floral meristem identity; LEAFY; callus; gene expression analysis
Issue or Number:2
Record Number:CaltechAUTHORS:20161205-100920077
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Official Citation:Wagner, D., Wellmer, F., Dilks, K., William, D., Smith, M. R., Kumar, P. P., Riechmann, J. L., Greenland, A. J. and Meyerowitz, E. M. (2004), Floral induction in tissue culture: a system for the analysis of LEAFY-dependent gene regulation. The Plant Journal, 39: 273–282. doi:10.1111/j.1365-313X.2004.02127.x
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:72549
Deposited By: Tony Diaz
Deposited On:07 Dec 2016 00:20
Last Modified:11 Nov 2021 05:03

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