Analysis of the floral transcriptome uncovers new regulators of organ determination and gene families related to flower organ differentiation in Gerbera hybrida (Asteraceae).

Development of composite inflorescences in the plant family Asteraceae has features that cannot be studied in the traditional model plants for flower development. In Gerbera hybrida, inflorescences are composed of morphologically different types of flowers tightly packed into a flower head (capitulum). Individual floral organs such as pappus bristles (sepals) are developmentally specialized, stamens are aborted in marginal flowers, petals and anthers are fused structures, and ovaries are located inferior to other floral organs. These specific features have made gerbera a rewarding target of comparative studies. Here we report the analysis of a gerbera EST database containing 16,994 cDNA sequences. Comparison of the sequences with all plant peptide sequences revealed 1656 unique sequences for gerbera not identified elsewhere within the plant kingdom. Based on the EST database, we constructed a cDNA microarray containing 9000 probes and have utilized it in identification of flower-specific genes and abundantly expressed marker genes for flower scape, pappus, stamen, and petal development. Our analysis revealed several regulatory genes with putative functions in flower-organ development. We were also able to associate a number of abundantly and specifically expressed genes with flower-organ differentiation. Gerbera is an outcrossing species, for which genetic approaches to gene discovery are not readily amenable. However, reverse genetics with the help of gene transfer has been very informative. We demonstrate here the usability of the gerbera microarray as a reliable new tool for identifying novel genes related to specific biological questions and for large-scale gene expression analysis.

[1]  N. Kumar,et al.  Role of polyamines and ethylene as modulators of plant senescence , 2000, Journal of Biosciences.

[2]  T. Teeri,et al.  Integration of reproductive meristem fates by a SEPALLATA-like MADS-box gene. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[3]  Elliot M. Meyerowitz,et al.  Genome-Wide Analysis of Spatial Gene Expression in Arabidopsis Flowers , 2004, The Plant Cell Online.

[4]  D. Soltis,et al.  Towards a comprehensive integration of morphological and genetic studies of floral development. , 2004, Trends in plant science.

[5]  Mark Stitt,et al.  Real-time RT-PCR profiling of over 1400 Arabidopsis transcription factors: unprecedented sensitivity reveals novel root- and shoot-specific genes. , 2004, The Plant journal : for cell and molecular biology.

[6]  J. Hawkins,et al.  Involvement of non-ABC MADS-box genes in determining stamen and carpel identity in Gerbera hybrida (Asteraceae) , 2004 .

[7]  R. Hall,et al.  Gene expression during anthesis and senescence in Iris flowers , 2003, Plant Molecular Biology.

[8]  Wei Hu,et al.  Isolation, sequence analysis, and expression studies of florally expressed cDNAs in Arabidopsis , 2003, Plant Molecular Biology.

[9]  D. Brummell,et al.  Cell wall metabolism in fruit softening and quality and its manipulation in transgenic plants , 2001, Plant Molecular Biology.

[10]  N. Chua,et al.  Molecular identification and characterization of the Arabidopsis AtADF1, AtADF5 and AtADF6 genes , 2001, Plant Molecular Biology.

[11]  S. Seo,et al.  Circadian and senescence-enhanced expression of a tobacco cysteine protease gene , 2000, Plant Molecular Biology.

[12]  U. Matern,et al.  Characterization and heterologous expression of hydroxycinnamoyl/benzoyl-CoA:anthranilate N-hydroxycinnamoyl/benzoyltransferase from elicited cell cultures of carnation, Dianthus caryophyllus L. , 1997, Plant Molecular Biology.

[13]  R. Dixon,et al.  Characterization of a gene encoding a DNA-binding protein that interacts in vitro with vascular specific cis elements of the phenylalanine ammonia-lyase promoter , 1997, Plant Molecular Biology.

[14]  A. Reddy,et al.  Cloning and expression of a PR5-like protein from Arabidopsis: inhibition of fungal growth by bacterially expressed protein , 1997, Plant Molecular Biology.

[15]  Y. Helariutta,et al.  Gerbera hybrida (Asteraceae) imposes regulation at several anatomical levels during inflorescence development on the gene for dihydroflavonol-4-reductase , 1995, Plant Molecular Biology.

[16]  L. Paulin,et al.  A corolla-and carpel-abundant, non-specific lipid transfer protein gene is expressed in the epidermis and parenchyma of Gerbera hybrida var. Regina (Compositae) , 1994, Plant Molecular Biology.

[17]  Y. Helariutta,et al.  Cloning of cDNA coding for dihydroflavonol-4-reductase (DFR) and characterization of dfr expression in the corollas of Gerbera hybrida var. Regina (Compositae) , 1993, Plant Molecular Biology.

[18]  S. McQueen-Mason,et al.  The molecular basis of plant cell wall extension , 2004, Plant Molecular Biology.

[19]  A. Steinmetz,et al.  A LIM-domain protein from sunflower is localized to the cytoplasm and/or nucleus in a wide variety of tissues and is associated with the phragmoplast in dividing cells , 2004, Plant Molecular Biology.

[20]  T. Teeri,et al.  Activation of Anthocyanin Biosynthesis in Gerbera hybrida (Asteraceae) Suggests Conserved Protein-Protein and Protein-Promoter Interactions between the Anciently Diverged Monocots and Eudicots1 , 2003, Plant Physiology.

[21]  Richard W McCombie,et al.  Expressed sequence tag analysis in Cycas, the most primitive living seed plant , 2003, Genome Biology.

[22]  J. Feijó,et al.  Transcriptional Profiling of Arabidopsis Tissues Reveals the Unique Characteristics of the Pollen Transcriptome1[w] , 2003, Plant Physiology.

[23]  Michael W. Frohlich,et al.  An evolutionary scenario for the origin of flowers , 2003, Nature Reviews Genetics.

[24]  D. Honys,et al.  Comparative Analysis of the Arabidopsis Pollen Transcriptome1[w] , 2003, Plant Physiology.

[25]  Richard G. H. Immink,et al.  The MADS Box Gene FBP2 Is Required for SEPALLATA Function in Petunia Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.010280. , 2003, The Plant Cell Online.

[26]  Hans-Werner Mewes,et al.  Sputnik: a database platform for comparative plant genomics , 2003, Nucleic Acids Res..

[27]  D. Honys,et al.  Comparative Analysis of the Arabidopsis Pollen Transcriptome , 2003 .

[28]  G. Jürgens,et al.  Microtubule cytoskeleton: a track record. , 2002, Current opinion in plant biology.

[29]  R. Dixon,et al.  A putative lipid transfer protein involved in systemic resistance signalling in Arabidopsis , 2002, Nature.

[30]  T. Ng,et al.  Isolation of an antifungal thaumatin-like protein from kiwi fruits. , 2002, Phytochemistry.

[31]  D. Ryan,et al.  Programmed cell death during flower senescence: isolation and characterization of cysteine proteinases from Sandersonia aurantiaca. , 2002, Functional plant biology : FPB.

[32]  Christine E. Horak,et al.  Global analysis of gene expression in yeast , 2002, Functional & Integrative Genomics.

[33]  David G Oppenheimer,et al.  Pleiotropy, redundancy and the evolution of flowers. , 2002, Trends in plant science.

[34]  D. Marion,et al.  From elicitins to lipid-transfer proteins: a new insight in cell signalling involved in plant defence mechanisms. , 2002, Trends in plant science.

[35]  C. Chapple,et al.  Rewriting the lignin roadmap. , 2002, Current opinion in plant biology.

[36]  Gerco C Angenent,et al.  Analysis of MADS box protein–protein interactions in living plant cells , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[37]  M. Purugganan,et al.  The genetics of plant morphological evolution. , 2002, Current opinion in plant biology.

[38]  T. Teeri,et al.  Involvement of non-ABC MADS-box genes in determining stamen and carpel indentity in Gerbera hybrida (Asteraceae) , 2002 .

[39]  D. Daly,et al.  Plant systematics in the age of genomics. , 2001, Plant physiology.

[40]  H. Ebinuma,et al.  Transcriptional control of lignin biosynthesis by tobacco LIM protein. , 2001, Phytochemistry.

[41]  Q. Cronk Plant evolution and development in a post-genomic context , 2001, Nature Reviews Genetics.

[42]  P. Facchini ALKALOID BIOSYNTHESIS IN PLANTS: Biochemistry, Cell Biology, Molecular Regulation, and Metabolic Engineering Applications. , 2001, Annual review of plant physiology and plant molecular biology.

[43]  A. Steinmetz,et al.  Molecular and expression analysis of a LIM protein gene family from flowering plants , 2000, Molecular Genetics and Genomics.

[44]  T. Teeri,et al.  GRCD1, an AGL2-like MADS Box Gene, Participates in the C Function during Stamen Development in Gerbera hybrida , 2000, Plant Cell.

[45]  A. Dunker,et al.  Isolation and characterization of cDNAs expressed in the early stages of flavonol-induced pollen germination in petunia. , 2000, Plant physiology.

[46]  K. Yoshida,et al.  Functional analysis of tobacco LIM protein Ntlim1 involved in lignin biosynthesis. , 2000, The Plant journal : for cell and molecular biology.

[47]  C. V. Jongeneel,et al.  ESTScan: A Program for Detecting, Evaluating, and Reconstructing Potential Coding Regions in EST Sequences , 1999, ISMB.

[48]  L. C. Loon,et al.  The families of pathogenesis-related proteins, their activities, and comparative analysis of PR-1 type proteins , 1999 .

[49]  Y. Helariutta,et al.  GEG Participates in the Regulation of Cell and Organ Shape during Corolla and Carpel Development in Gerbera hybrida , 1999, Plant Cell.

[50]  Y. Helariutta,et al.  Organ identity genes and modified patterns of flower development in Gerbera hybrida (Asteraceae) , 1999, The Plant journal : for cell and molecular biology.

[51]  Y. Helariutta,et al.  Flower development and secondary metabolism in Gerbera hybrida, an Asteraceae , 1999 .

[52]  Y. Helariutta,et al.  New pathway to polyketides in plants , 1998, Nature.

[53]  Y. Helariutta,et al.  A bHLH transcription factor mediates organ, region and flower type specific signals on dihydroflavonol-4-reductase (dfr) gene expression in the inflorescence of Gerbera hybrida (Asteraceae). , 1998, The Plant journal : for cell and molecular biology.

[54]  B. Weisshaar,et al.  Phenylpropanoid biosynthesis and its regulation. , 1998, Current opinion in plant biology.

[55]  J. Strommer,et al.  Myb26: a MYB-like protein of pea flowers with affinity for promoters of phenylpropanoid genes. , 1997, The Plant journal : for cell and molecular biology.

[56]  J. Kader Lipid-transfer proteins: a puzzling family of plant proteins , 1997 .

[57]  J. V. van Kan,et al.  Cutinase A of Botrytis cinerea is expressed, but not essential, during penetration of gerbera and tomato. , 1997, Molecular plant-microbe interactions : MPMI.

[58]  C. Martin,et al.  Apparent redundancy in myb gene function provides gearing for the control of flavonoid biosynthesis in antirrhinum flowers. , 1996, The Plant cell.

[59]  Y. Helariutta,et al.  Duplication and functional divergence in the chalcone synthase gene family of Asteraceae: evolution with substrate change and catalytic simplification. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[60]  K. Bremer,et al.  Asteraceae: Cladistics and Classification , 1994 .

[61]  M. Bevan,et al.  A flower‐specific Myb protein activates transcription of phenylpropanoid biosynthetic genes. , 1994, The EMBO journal.

[62]  A. Steinmetz,et al.  A LIM motif is present in a pollen-specific protein. , 1992, The Plant cell.

[63]  H. Woodland,et al.  Histone genes: Not so simple after all , 1984, Cell.

[64]  J. Mansfield,et al.  Mode of Action of Pollen in Breaking Resistance of Vicia faba to Botrytis cinerea , 1971, Nature.

[65]  T. Preece,et al.  The effect of pollen grains on infections caused by Botrytis cinerea Fr. , 1968, The Annals of applied biology.