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First published online October 23, 2003; 10.1105/tpc.017376

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The Plant Cell, Vol. 15, 2680-2693, November 2003, www.plantcell.org ©2003, American Society of Plant Biologists

Toward the Analysis of the Petunia MADS Box Gene Family by Reverse and Forward Transposon Insertion Mutagenesis Approaches: B, C, and D Floral Organ Identity Functions Require SEPALLATA-Like MADS Box Genes in Petunia

Michiel Vandenbusschea, Jan Zethofa, Erik Souer1,b, Ronald Koesb, Giovanni B. Torniellic, Mario Pezzottic, Silvia Ferrariod, Gerco C. Angenentd and Tom Gerats2,3,a

a Department of Plant Systems Biology, Vlaams Instituut voor Biotechnologie/Ghent University, B-9052 Zwijnaarde, Belgium
b Department of Genetics, Vrije Universiteit Amsterdam, 1081 HV Amsterdam, The Netherlands
c Dipartimento Scientifico e Tecnologico, Università degli Studi di Verona, 37134 Verona, Italy
d Business Unit Bioscience, Plant Research International, 6700 AA Wageningen, The Netherlands

3 To whom correspondence should be addressed. E-mail tgerats{at}sci.kun.nl; fax 31-243-553450

We have initiated a systematic functional analysis of the MADS box, intervening region, K domain, C domain-type MADS box gene family in petunia. The starting point for this has been a reverse-genetics approach, aiming to select for transposon insertions into any MADS box gene. We have developed and applied a family signature insertion screening protocol that is highly suited for this purpose, resulting in the isolation of 32 insertion mutants in 20 different MADS box genes. In addition, we identified three more MADS box gene insertion mutants using a candidate-gene approach. The defined insertion lines provide a sound foundation for a systematic functional analysis of the MADS box gene family in petunia. Here, we focus on the analysis of Floral Binding Protein2 (FBP2) and FBP5 genes that encode the E-function, which in Arabidopsis has been shown to be required for B and C floral organ identity functions. fbp2 mutants display sepaloid petals and ectopic inflorescences originating from the third floral whorl, whereas fbp5 mutants appear as wild type. In fbp2 fbp5 double mutants, reversion of floral organs to leaf-like organs is increased further. Strikingly, ovules are replaced by leaf-like structures in the carpel, indicating that in addition to the B- and C-functions, the D-function, which specifies ovule development, requires E-function activity. Finally, we compare our data with results obtained using cosuppression approaches and conclude that the latter might be less suited for assigning functions to individual members of the MADS box gene family.




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