Advertisement

Planta

, 228:401 | Cite as

Green corolla segments in a wild Petunia species caused by a mutation in FBP2, a SEPALLATA-like MADS box gene

  • Kiyoshi Matsubara
  • Katsuyoshi Shimamura
  • Hiroaki Kodama
  • Hisashi Kokubun
  • Hitoshi Watanabe
  • Isabel L. Basualdo
  • Toshio Ando
Original Article

Abstract

A Petunia inflata isolate with a novel phenotype of a purple corolla limb with green corolla segments (GCS) was characterized. The GCS have stomata and trichomes on the adaxial side, and resemble calyx segments in epidermal morphology. The GCS phenotype was inherited in a recessive manner. In the GCS plant, a novel inhibitor/defective spm-like transposable element (dPifTp1) was inserted in the second intron of the Floral Binding Protein 2 (FBP2) gene. The sequence of the resulting transcript contained five silent mutations as compared the corresponding open reading frame of P. × hybrida FBP2 mRNA. The GCS phenotype co-segregated with an FBP2 fragment containing a dPifTp1 insertion. The transcript level of the FBP2 gene in GCS flowers was markedly lower than that in wild-type (WT) flowers, suggesting that partially inhibited FBP2 gene expression caused the morphogenesis of calyx-like tissue in the corolla segments of GCS flowers. Gene expression pattern analysis using a full-length Petunia floral cDNA microarray indicated that some photosynthesis-related genes were expressed at significantly higher levels in the GCS of GCS flowers, but the mRNA levels of most other genes in the GCS were similar to those in the WT corolla. Taken together, these data suggest that the partial loss of FBP2 expression does not shift global gene expression in the corolla segments of the GCS flower toward that of calyx, even though calyx-like morphogenesis was established in the corolla segments.

Keywords

cDNA microarray FBP2 gene Floral morphology MADS box Petunia Transposon 

Abbreviations

FBP2

Floral Binding Protein 2

GCS

Green corolla segments

spm

Suppressor mutator

WT

Wild type

Supplementary material

425_2008_744_MOESM1_ESM.tif (205 kb)
PCR analysis of DNA isolated from P. inflata WT (lane 1), GCS (lane 2), and F1 (lane 3) and F2 (lane 4 to 6) generations produced by crossing a WT plant and a GCS plant. The DNA fragment was amplified with the primers fbp2-f3 (5’-GCACCAGAGACTAATATATCCACACG-3’) and fbp2-r3 (5’-TTGGCTGCTTATTTCCTGTAATCAT-3’). The fragment length is associated with the alleles FBP2 (1.7 kbp) and fbp2 with dPifTp1 (3.8 kbp). Phenotypes and generations are shown below each lane. The numbers to the left indicate DNA size markers (in kb) (TIFF 205 kb)
425_2008_744_MOESM2_ESM.tif (178 kb)
Scatter plots of expression distribution patterns of 2,976 ESTs after microarray hybridization with labelled cDNA probes obtained from mRNAs of four different floral tissues: GCS limb (purple corolla limb of GCS mutant), GCS segment (green corolla segments of GCS mutant), WT corolla (purple corolla segments plus purple corolla limb of WT), and WT calyx (calyx segments of WT). The diagonal lines indicate 1.5-fold relative intensity differences between the two RNA samples. On each diagram, the FBP2 gene is represented by solid triangles. The genes upregulated in GCS segment and WT calyx as compared to WT corolla are represented by open triangles. (a) Distribution of the signal log ratio values for WT corolla (x axis) and GCS limb (y axis). (b) Distribution of the signal log ratio values for WT corolla (x axis) and GCS segment (y axis). (c) Distribution of the signal log ratio values for WT corolla (x axis) and WT calyx (y axis) (TIFF 178 kb)

References

  1. Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402PubMedCrossRefGoogle Scholar
  2. Ando T, Hashimoto G (1993) Two new species of Petunia (Solanaceae) from southern Brazil. Bot J Linn Soc 111:265–280CrossRefGoogle Scholar
  3. Angenent GC, Busscher M, Franken J, Mol JNM, Van Tunen AJ (1992) Differential expression of two MADS box genes in wild-type and mutant petunia flowers. Plant Cell 4:983–993PubMedCrossRefGoogle Scholar
  4. Angenent GC, Franken J, Busscher M, Weiss D, Van Tunen AJ (1994) Co-suppression of the petunia homeotic gene fbp2 affects the identity of the generative meristem. Plant J 5:33–44PubMedCrossRefGoogle Scholar
  5. Bailey LH (1896) Evolution of the Petunia. In: The survival of the unlike, Macmillan, London, pp 465–472Google Scholar
  6. Esau K (1977) Anatomy of seed plants, 2nd edn. Wiley, New YorkGoogle Scholar
  7. Fedoroff NV (1989) Maize transposable elements. In: Howe M, Berg P (eds) Mobile DNA. American Society for Microbiology, Washington, pp 375–411Google Scholar
  8. Ferrario S, Immink RG, Shchennikova A, Busscher-Lange J, Angenent GC (2003) The MADS box gene FBP2 is required for SEPALLATA function in petunia. Plant Cell 15:914–925PubMedCrossRefGoogle Scholar
  9. Hirai S, Oka S, Adachi E, Kodama H (2007) The effects of spacer sequences on silencing efficiency of plant RNAi vectors. Plant Cell Rep 26:651–659PubMedCrossRefGoogle Scholar
  10. Jernstedt UA, Cutter EG, Gifford EM, Lu P (1992) Angle meristem origin and development in Selaginella martensii. Ann Bot 69:351–363Google Scholar
  11. Kodama H, Hamada T, Horiguchi G, Nishimura M, Iba K (1994) Genetic enhancement of cold tolerance by expression of a gene for chloroplast omega-3 fatty acid desaturase in transgenic tobacco. Plant Physiol 105:601–605PubMedGoogle Scholar
  12. Lassner MW, Peterson P, Yoder JI (1989) Simultaneous amplification of multiple DNA fragments by polymerase chain reaction in the analysis of transgenic plants and their progeny. Plant Mol Biol Rep 7:116–128CrossRefGoogle Scholar
  13. Nagira Y, Shimamura K, Hirai S, Shimanuki M, Kodama H, Ozeki Y (2006) Identification and characterization of genes induced for anthocyanin synthesis and chlorophyll degradation in regenerated torenia shoots using suppression subtractive hybridization, cDNA microarrays, and RNAi techniques. J Plant Res 119:217–230PubMedCrossRefGoogle Scholar
  14. Paxton J (1836) Petunia nyctaginiflora violacea. Paxton’s Mag Bot 2:173Google Scholar
  15. Pelaz S, Ditta GS, Baumann E, Wisman E, Yanofsky MF (2000) B and C floral organ identity functions require SEPALLATA MADS box genes. Nature 405:200–203PubMedCrossRefGoogle Scholar
  16. Shimamura K, Ishimizu T, Nishimura K, Matsubara K, Kodama H, Watanabe H, Hase S, Ando T (2007) Analysis of expressed sequence tags from Petunia flowers. Plant Sci 173:495–500CrossRefGoogle Scholar
  17. Shimanuki M, Shimamura K, Hirai S, Nishiuchi T, Suzuki K, Kodama H (2005) Polyethylene glycol-mediated enhancement of the hybridization rate on cDNA microarrays. Anal Biochem 344:284–286PubMedGoogle Scholar
  18. Vandenbussche M, Zethof J, Souer E, Koes R, Tornielli GB, Pezzotti M, Ferrario S, Angenent GC, Gerats T (2003) 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. Plant Cell 15:2680–2693PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Kiyoshi Matsubara
    • 1
  • Katsuyoshi Shimamura
    • 1
  • Hiroaki Kodama
    • 2
  • Hisashi Kokubun
    • 3
  • Hitoshi Watanabe
    • 3
  • Isabel L. Basualdo
    • 4
  • Toshio Ando
    • 2
  1. 1.Graduate School of Science and TechnologyChiba UniversityChibaJapan
  2. 2.Graduate School of HorticultureChiba UniversityChibaJapan
  3. 3.Center for Environment, Health and Field SciencesChiba UniversityKashiwa, ChibaJapan
  4. 4.Sección Botánica, Facultad de Ciencias QuímicasUniversidad Nacional de AsunciónSan LorenzoParaguay

Personalised recommendations