An R2R3-MYB Transcription Factor Regulates Eugenol Production in Ripe Strawberry Fruit Receptacles1

A ripening-related transcription factor affects the phenylpropanoid synthesis pathway and the biosynthesis of eugenol in the strawberry fruit. Eugenol is a volatile phenylpropanoid that contributes to flower and ripe fruit scent. In ripe strawberry (Fragaria × ananassa) fruit receptacles, eugenol is biosynthesized by eugenol synthase (FaEGS2). However, the transcriptional regulation of this process is still unknown. We have identified and functionally characterized an R2R3 MYB transcription factor (EMISSION OF BENZENOID II [FaEOBII]) that seems to be the orthologous gene of PhEOBII from Petunia hybrida, which contributes to the regulation of eugenol biosynthesis in petals. The expression of FaEOBII was ripening related and fruit receptacle specific, although high expression values were also found in petals. This expression pattern of FaEOBII correlated with eugenol content in both fruit receptacle and petals. The expression of FaEOBII was repressed by auxins and activated by abscisic acid, in parallel to the ripening process. In ripe strawberry receptacles, where the expression of FaEOBII was silenced, the expression of CINNAMYL ALCOHOL DEHYDROGENASE1 and FaEGS2, two structural genes involved in eugenol production, was down-regulated. A subsequent decrease in eugenol content in ripe receptacles was also observed, confirming the involvement of FaEOBII in eugenol metabolism. Additionally, the expression of FaEOBII was under the control of FaMYB10, another R2R3 MYB transcription factor that regulates the early and late biosynthetic genes from the flavonoid/phenylpropanoid pathway. In parallel, the amount of eugenol in FaMYB10-silenced receptacles was also diminished. Taken together, these data indicate that FaEOBII plays a regulating role in the volatile phenylpropanoid pathway gene expression that gives rise to eugenol production in ripe strawberry receptacles.

[1]  W. Schwab,et al.  MYB10 plays a major role in the regulation of flavonoid/phenylpropanoid metabolism during ripening of Fragaria x ananassa fruits. , 2014, Journal of experimental botany.

[2]  J. Franco-Zorrilla,et al.  DNA-binding specificities of plant transcription factors and their potential to define target genes , 2014, Proceedings of the National Academy of Sciences.

[3]  W. Schwab,et al.  Eugenol Production in Achenes and Receptacles of Strawberry Fruits Is Catalyzed by Synthases Exhibiting Distinct Kinetics1[W][OPEN] , 2013, Plant Physiology.

[4]  G. Casadoro,et al.  A SHATTERPROOF-like gene controls ripening in non-climacteric strawberries, and auxin and abscisic acid antagonistically affect its expression , 2013, Journal of experimental botany.

[5]  Hideyuki Suzuki,et al.  Enhancement of production of eugenol and its glycosides in transgenic aspen plants via genetic engineering. , 2013, Biochemical and biophysical research communications.

[6]  A. Monfort,et al.  The strawberry (Fragariaxananassa) fruit-specific rhamnogalacturonate lyase 1 (FaRGLyase1) gene encodes an enzyme involved in the degradation of cell-wall middle lamellae. , 2013, Journal of experimental botany.

[7]  B. Spitzer-Rimon,et al.  The R2R3-MYB–Like Regulatory Factor EOBI, Acting Downstream of EOBII, Regulates Scent Production by Activating ODO1 and Structural Scent-Related Genes in Petunia[C][W] , 2012, Plant Cell.

[8]  W. Schwab,et al.  The fruit ripening-related gene FaAAT2 encodes an acyl transferase involved in strawberry aroma biogenesis. , 2012, Journal of experimental botany.

[9]  A. Granell,et al.  Genetic Analysis of Strawberry Fruit Aroma and Identification of O-Methyltransferase FaOMT as the Locus Controlling Natural Variation in Mesifurane Content1[C][W][OA] , 2012, Plant Physiology.

[10]  Yuan-Yue Shen,et al.  FaPYR1 is involved in strawberry fruit ripening. , 2011, Journal of experimental botany.

[11]  Robert C Schuurink,et al.  The transcription factor EMISSION OF BENZENOIDS II activates the MYB ODORANT1 promoter at a MYB binding site specific for fragrant petunias. , 2011, The Plant journal : for cell and molecular biology.

[12]  E. Pichersky,et al.  Metabolic engineering in strawberry fruit uncovers a dormant biosynthetic pathway. , 2011, Metabolic engineering.

[13]  Yuan-Yue Shen,et al.  Abscisic Acid Plays an Important Role in the Regulation of Strawberry Fruit Ripening1[W][OA] , 2011, Plant Physiology.

[14]  R. McQuinn,et al.  Identification of Genes in the Phenylalanine Metabolic Pathway by Ectopic Expression of a MYB Transcription Factor in Tomato Fruit[W] , 2011, Plant Cell.

[15]  J. Franco-Zorrilla,et al.  Improved protein-binding microarrays for the identification of DNA-binding specificities of transcription factors. , 2011, The Plant journal : for cell and molecular biology.

[16]  T. Colquhoun,et al.  Unraveling the regulation of floral fragrance biosynthesis , 2011, Plant signaling & behavior.

[17]  Henry D. Priest,et al.  The genome of woodland strawberry (Fragaria vesca) , 2011, Nature Genetics.

[18]  J. Y. Kim,et al.  PhMYB4 fine-tunes the floral volatile signature of Petunia×hybrida through PhC4H , 2010, Journal of experimental botany.

[19]  A. Bombarely,et al.  Generation and analysis of ESTs from strawberry (Fragaria xananassa) fruits and evaluation of their utility in genetic and molecular studies , 2010, BMC Genomics.

[20]  J. McCarthy,et al.  Acaricidal Activity of Eugenol Based Compounds against Scabies Mites , 2010, PloS one.

[21]  P. Perkins-Veazie Growth and Ripening of Strawberry Fruit , 2010 .

[22]  Zhangjun Fei,et al.  Transcriptional profiles of drought-responsive genes in modulating transcription signal transduction, and biochemical pathways in tomato , 2010, Journal of experimental botany.

[23]  B. Spitzer-Rimon,et al.  EOBII, a Gene Encoding a Flower-Specific Regulator of Phenylpropanoid Volatiles' Biosynthesis in Petunia[C][W] , 2010, Plant Cell.

[24]  Beverly A. Underwood,et al.  Petunia floral volatile benzenoid/phenylpropanoid genes are regulated in a similar manner. , 2010, Phytochemistry.

[25]  J. Gil,et al.  Quantitative comparison of free and bound volatiles of two commercial tomato cultivars (Solanum lycopersicum L.) during ripening. , 2010, Journal of agricultural and food chemistry.

[26]  R. Thornburg,et al.  The MYB305 Transcription Factor Regulates Expression of Nectarin Genes in the Ornamental Tobacco Floral Nectary[C][W] , 2009, The Plant Cell Online.

[27]  Claudia A Bustamante,et al.  Cloning of the promoter region of β-xylosidase (FaXyl1) gene and effect of plant growth regulators on the expression of FaXyl1 in strawberry fruit , 2009 .

[28]  A. Maldonado,et al.  Evidence for a positive regulatory role of strawberry (Fragaria x ananassa) Fa WRKY1 and Arabidopsis At WRKY75 proteins in resistance. , 2009, Journal of experimental botany.

[29]  M. Berger,et al.  Universal protein-binding microarrays for the comprehensive characterization of the DNA-binding specificities of transcription factors , 2009, Nature Protocols.

[30]  W. Schwab,et al.  Multi-substrate flavonol O-glucosyltransferases from strawberry (Fragaria×ananassa) achene and receptacle , 2008, Journal of experimental botany.

[31]  M. Bowman,et al.  The multiple phenylpropene synthases in both Clarkia breweri and Petunia hybrida represent two distinct protein lineages. , 2008, The Plant journal : for cell and molecular biology.

[32]  Mark A. Matthews,et al.  Water deficits accelerate ripening and induce changes in gene expression regulating flavonoid biosynthesis in grape berries , 2007, Planta.

[33]  J. Glazebrook,et al.  In planta transformation of Arabidopsis. , 2006, CSH protocols.

[34]  W. Schwab,et al.  RNAi-induced silencing of gene expression in strawberry fruit (Fragaria x ananassa) by agroinfiltration: a rapid assay for gene function analysis. , 2006, The Plant journal : for cell and molecular biology.

[35]  M. Pitrat,et al.  Volatile compounds in the skin and pulp of Queen Anne's pocket melon. , 2006, Journal of agricultural and food chemistry.

[36]  J. Noel,et al.  Eugenol and isoeugenol, characteristic aromatic constituents of spices, are biosynthesized via reduction of a coniferyl alcohol ester. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[37]  R. Simpson Bulk precipitation of proteins by ammonium sulfate. , 2006, CSH protocols.

[38]  Jigang Li,et al.  Two groups of MYB transcription factors share a motif which enhances trans-activation activity. , 2006, Biochemical and biophysical research communications.

[39]  W. Schwab,et al.  FaQR, Required for the Biosynthesis of the Strawberry Flavor Compound 4-Hydroxy-2,5-Dimethyl-3(2H)-Furanone, Encodes an Enone Oxidoreductase , 2006, The Plant Cell Online.

[40]  M. Haring,et al.  ODORANT1 Regulates Fragrance Biosynthesis in Petunia Flowersw⃞ , 2005, The Plant Cell Online.

[41]  M. Bellido,et al.  A strawberry fruit-specific and ripening-related gene codes for a HyPRP protein involved in polyphenol anchoring , 2004, Plant Molecular Biology.

[42]  D. Bedgar,et al.  Crystal Structures of Pinoresinol-Lariciresinol and Phenylcoumaran Benzylic Ether Reductases and Their Relationship to Isoflavone Reductases* , 2003, Journal of Biological Chemistry.

[43]  Robert C Schuurink,et al.  Regulation of floral scent production in petunia revealed by targeted metabolomics. , 2003, Phytochemistry.

[44]  D. Baulcombe,et al.  An enhanced transient expression system in plants based on suppression of gene silencing by the p19 protein of tomato bushy stunt virus. , 2003, The Plant journal : for cell and molecular biology.

[45]  A. Benítez-Burraco,et al.  Cloning and characterization of two ripening-related strawberry (Fragaria x ananassa cv. Chandler) pectate lyase genes. , 2003, Journal of experimental botany.

[46]  N. Medina-Escobar,et al.  Cloning, expression and immunolocalization pattern of a cinnamyl alcohol dehydrogenase gene from strawberry (Fragaria x ananassa cv. Chandler). , 2002, Journal of experimental botany.

[47]  Dirk Inzé,et al.  GATEWAY vectors for Agrobacterium-mediated plant transformation. , 2002, Trends in plant science.

[48]  G. Choi,et al.  AtMYB21, a gene encoding a flower-specific transcription factor, is regulated by COP1. , 2002, The Plant journal : for cell and molecular biology.

[49]  E. Grotewold,et al.  MYB transcription factors in Arabidopsis. , 2002, Trends in plant science.

[50]  A. Aharoni,et al.  The strawberry FaMYB1 transcription factor suppresses anthocyanin and flavonol accumulation in transgenic tobacco. , 2001, The Plant journal : for cell and molecular biology.

[51]  M. Jordán,et al.  Aromatic profile of aqueous banana essence and banana fruit by gas chromatography-mass spectrometry (GC-MS) and gas chromatography-olfactometry (GC-O). , 2001, Journal of agricultural and food chemistry.

[52]  T. Ruttink,et al.  The use of the luciferase reporter system forin planta gene expression studies , 2000, Plant Molecular Biology Reporter.

[53]  P. Dhawan,et al.  A simple procedure for the isolation of high quality RNA from ripening banana fruit , 2000, Plant Molecular Biology Reporter.

[54]  G. Casadoro,et al.  A novel E-type endo-β-1,4-glucanase with a putative cellulose-binding domain is highly expressed in ripening strawberry fruits , 1999, Plant Molecular Biology.

[55]  C. Tonelli,et al.  Towards functional characterisation of the members of the R2R3-MYB gene family from Arabidopsis thaliana. , 1998, The Plant journal : for cell and molecular biology.

[56]  K. Manning Isolation of a set of ripening-related genes from strawberry: their identification and possible relationship to fruit quality traits , 1998, Planta.

[57]  V. Valpuesta,et al.  A fruit-specific putative dihydroflavonol 4-reductase gene is differentially expressed in strawberry during the ripening process. , 1998, Plant physiology.

[58]  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.

[59]  N. Medina-Escobar,et al.  Cloning, molecular characterization and expression pattern of a strawberry ripening-specific cDNA with sequence homology to pectate lyase from higher plants , 1997, Plant Molecular Biology.

[60]  I. Zabetakis,et al.  Strawberry Flavour: Analysis and Biosynthesis , 1997 .

[61]  P. Schieberle,et al.  Evaluation of the character impact odorants in fresh strawberry juice by quantitative measurements and sensory studies on model mixtures , 1997 .

[62]  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.

[63]  A. Rapp,et al.  Analyse des Erdbeeraromas - Quantifizierung der flüchtigen Komponenten in Kulturerdbeervarietäten und der Walderdbeere , 1995 .

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

[65]  R. Creelman,et al.  Involvement of a lipoxygenase-like enzyme in abscisic Acid biosynthesis. , 1992, Plant physiology.

[66]  R. A. Ludwig,et al.  A DNA Transformation–Competent Arabidopsis Genomic Library in Agrobacterium , 1991, Bio/Technology.

[67]  Leif Poll,et al.  Odour thresholds of some important aroma compounds in strawberries , 1990 .

[68]  J. R. Woodward Physical and chemical changes in developing strawberry fruits. , 1972, Journal of the science of food and agriculture.

[69]  M. Campbell,et al.  The interaction between MYB proteins and their target DNA binding sites. , 2012, Biochimica et biophysica acta.

[70]  D. Reinhardt,et al.  A petunia chorismate mutase specialized for the production of floral volatiles. , 2010, The Plant journal : for cell and molecular biology.

[71]  E. Pichersky,et al.  Characterization of a petunia acetyltransferase involved in the biosynthesis of the floral volatile isoeugenol. , 2007, The Plant journal : for cell and molecular biology.

[72]  D. Ulrich,et al.  Diversity of aroma patterns in wild and cultivated Fragaria accessions , 2006, Genetic Resources and Crop Evolution.

[73]  M. Haring,et al.  Regulation of volatile benzenoid biosynthesis in petunia flowers. , 2006, Trends in plant science.

[74]  D. Joyce,et al.  ABA effects on ethylene production, PAL activity, anthocyanin and phenolic contents of strawberry fruit , 2004, Plant Growth Regulation.

[75]  Yoshihiro Ugawa,et al.  Plant cis-acting regulatory DNA elements (PLACE) database: 1999 , 1999, Nucleic Acids Res..

[76]  Y. Benjamini,et al.  Controlling the false discovery rate: a practical and powerful approach to multiple testing , 1995 .

[77]  H Maarse,et al.  Volatile compounds in food : qualitative and quantitative data , 1990 .

[78]  Timo Hirvi,et al.  Volatiles of wild strawberries, Fragaria vesca L., compared to those of cultivated berries, Fragaria .times. ananassa cv Senga Sengana , 1979 .