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Two highly divergent alcohol dehydrogenases of melon exhibit fruit ripening-specific expression and distinct biochemical characteristics

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Abstract

Alcohol dehydrogenases (ADH) participate in the biosynthetic pathway of aroma volatiles in fruit by interconverting aldehydes to alcohols and providing substrates for the formation of esters. Two highly divergent ADH genes (15% identity at the amino acid level) of Cantaloupe Charentais melon (Cucumis melo var. Cantalupensis) have been isolated. Cm-ADH1 belongs to the medium-chain zinc-binding type of ADHs and is highly similar to all ADH genes expressed in fruit isolated so far. Cm-ADH2 belongs to the short-chain type of ADHs. The two encoded proteins are enzymatically active upon expression in yeast. Cm-ADH1 has strong preference for NAPDH as a co-factor, whereas Cm-ADH2 preferentially uses NADH. Both Cm-ADH proteins are much more active as reductases with K ms 10–20 times lower for the conversion of aldehydes to alcohols than for the dehydrogenation of alcohols to aldehydes. They both show strong preference for aliphatic aldehydes but Cm-ADH1 is capable of reducing branched aldehydes such as 3-methylbutyraldehyde, whereas Cm-ADH2 cannot. Both Cm-ADH genes are expressed specifically in fruit and up-regulated during ripening. Gene expression as well as total ADH activity are strongly inhibited in antisense ACC oxidase melons and in melon fruit treated with the ethylene antagonist 1-methylcyclopropene (1-MCP), indicating a positive regulation by ethylene. These data suggest that each of the Cm-ADH protein plays a specific role in the regulation of aroma biosynthesis in melon fruit.

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References

  • R Ayub M Guis M Ben Amor L Gillot JP Roustan A Latché M Bouzayen JC Pech (1996) ArticleTitleExpression of ACC oxidase antisense gene inhibits ripening of cantaloupe melon fruits Nat Biotech 14 862–866 Occurrence Handle1:CAS:528:DyaK28XktF2ktrg%3D Occurrence Handle10.1038/nbt0796-862

    Article  CAS  Google Scholar 

  • SF Altschul TL Madden AA Schaffer J Zhang Z Zhang W Miller DJ Lipman (1997) ArticleTitleGapped BLAST and PSI-BLAST: a new generation of protein database search programs Nucleic Acids Res 25 3389–3402 Occurrence Handle9254694 Occurrence Handle1:CAS:528:DyaK2sXlvFyhu7w%3D Occurrence Handle10.1093/nar/25.17.3389

    Article  PubMed  CAS  Google Scholar 

  • C Aubert N Bourger (2004) ArticleTitleInvestigation of volatiles in Charentais Cantaloupe melons (Cucumis melo Var Cantalupensis). Characterization of aroma constituents in some cultivars J Agric Food Chem 52 4522–4528 Occurrence Handle15237961 Occurrence Handle1:CAS:528:DC%2BD2cXkvVWqs7w%3D Occurrence Handle10.1021/jf049777s

    Article  PubMed  CAS  Google Scholar 

  • PK Boss C Davies SP Robinson (1996) ArticleTitleAnalysis of the expression of anthocyanin pathway genes in developing Vitis vinifera cv.Shiraz grape berries and the implications for pathway regulation Plant Physiol 111 1059–1066 Occurrence Handle12226348 Occurrence Handle1:CAS:528:DyaK28XltVGgt78%3D

    PubMed  CAS  Google Scholar 

  • MM Bradford (1976) ArticleTitleA rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein–dye binding Anal Biochem 72 248–254 Occurrence Handle942051 Occurrence Handle1:CAS:528:DyaE28XksVehtrY%3D Occurrence Handle10.1016/0003-2697(76)90527-3

    Article  PubMed  CAS  Google Scholar 

  • T Chase SuffixJr (1999) ArticleTitleAlcohol dehydrogenases: identification and names for gene families Plant Mol Biol Rep 17 333–350 Occurrence Handle1:CAS:528:DC%2BD3cXjvFSmurg%3D Occurrence Handle10.1023/A:1007620627083

    Article  CAS  Google Scholar 

  • A-RS Chen T Chase SuffixJr (1993) ArticleTitleAlcohol dehydrogenase 2 and pyruvate decarboxylase induction in ripening and hypoxy tomato fruit Plant Physiol 31 875–885 Occurrence Handle1:CAS:528:DyaK2cXisVShtr8%3D

    CAS  Google Scholar 

  • GL Bruxelles ParticleDe WJ Peacock ES Dennis R Dolferus (1996) ArticleTitleAbscisic acid induces the alcohol dehydrogenase gene in Arabidopsis Plant Physiol 111 381–391 Occurrence Handle8787023 Occurrence Handle10.1104/pp.111.2.381

    Article  PubMed  Google Scholar 

  • G Echeverria J Graell ML Lopez I Lara (2004) ArticleTitleVolatile production, quality, and aroma-related enzyme activities during maturation of “Fuji” apples Postharvest Biol Technol 31 217–227 Occurrence Handle1:CAS:528:DC%2BD2cXhslCitLk%3D Occurrence Handle10.1016/j.postharvbio.2003.09.003

    Article  CAS  Google Scholar 

  • H Eklund B Nordström E Zeppezauer G Söderlund I Ohlsson T Boiwe BO Söderberg O Tapia CI Brändén Å Åkeson (1976) ArticleTitleThree-dimensional structure of horse liver alcohol dehydrogenase at 2.4 Å resolution J Mol Biol 102 27–59 Occurrence Handle178875 Occurrence Handle1:CAS:528:DyaE28Xhs1Wit7c%3D Occurrence Handle10.1016/0022-2836(76)90072-3

    Article  PubMed  CAS  Google Scholar 

  • I El-Sharkawy D Manríquez FB Flores F Regad M Bouzayen A Latché JC Pech (2005) ArticleTitleFunctional characterization of a melon alcohol acyl-transferase gene family involved in the biosynthesis of ester volatiles. Identification of the crucial role of a threonine residue for enzyme activity Plant Mol Biol 59 343–360 Occurrence Handle10.1007/s11103-005-8884-y Occurrence Handle1:CAS:528:DC%2BD2MXhtFCrtLfN

    Article  CAS  Google Scholar 

  • F Fan JA Lorenzen BV Plapp (1991) ArticleTitleAn aspartate residue in yeast alcohol dehydrogenase I determines the specificity for coenzyme Biochemistry 30 6397–6401 Occurrence Handle2054345 Occurrence Handle1:CAS:528:DyaK3MXkt1Cqtr4%3D Occurrence Handle10.1021/bi00240a008

    Article  PubMed  CAS  Google Scholar 

  • Felsenstein J (1992) PHYLIP (Univ. of Washington, Seattle), Version 3.5

  • S Fonseca L Hackler SuffixJr A Zvara S Ferreira A Baldé D Dudits MS Pais LG Puskas (2004) ArticleTitleMonitoring gene expression along pear fruit development, ripening and senescence using cDNA microarrays Plant Sci 167 457–469 Occurrence Handle1:CAS:528:DC%2BD2cXlsFOqt7o%3D Occurrence Handle10.1016/j.plantsci.2004.03.033

    Article  CAS  Google Scholar 

  • F Flores F El Yahyaoui G Billerbeck Particlede F Romojaro A Latché M Bouzayen JC Pech C Ambid (2002) ArticleTitleRole of ethylene in the biosynthetic pathway of aliphatic ester aroma volatiles in Charentais Cantaloupe melons J Exp Bot 53 201–206 Occurrence Handle11807123 Occurrence Handle1:CAS:528:DC%2BD38XhtlWkt7g%3D Occurrence Handle10.1093/jexbot/53.367.201

    Article  PubMed  CAS  Google Scholar 

  • M Guis R Botondi M Ben Amor R Ayub M Bouzayen JC Pech A Latché (1997) ArticleTitleRipening-associated biochemical traits of cantaloupe charentais melons expressing an antisense ACC oxidase transgene J Am Soc Hort Sci 122 748–751 Occurrence Handle1:CAS:528:DyaK2sXntlOlt7g%3D

    CAS  Google Scholar 

  • JC Ingersoll M Rothenberg BE Liedl K Folkerts D Garvin MR Hanson JJ Doyle MA Mutschler (1994) ArticleTitleA novel anther-expressed adh-homologous gene in Lycopersicon esculentum Plant Mol Biol 26 1875–1891 Occurrence Handle7858224 Occurrence Handle1:CAS:528:DyaK2MXktV2gtLc%3D Occurrence Handle10.1007/BF00019500

    Article  PubMed  CAS  Google Scholar 

  • H Jörnvall B Persson M Krook S Atrian R Gonzalez-Duarte J Jeffery D Ghosh (1995) ArticleTitleShort-chain dehydrogenases/reductases (SDR) Biochemistry 34 6003–6013 Occurrence Handle7742302 Occurrence Handle10.1021/bi00018a001

    Article  PubMed  Google Scholar 

  • B Klein K Pawlowski C Hoericke-Grandpierre J Schell R Toepfer (1992) ArticleTitleIsolation and characterization of a cDNA from Cuphea lanceolata encoding a beta-ketoacyl-ACP reductase Mol Gen Genet 233 122–128 Occurrence Handle1376402 Occurrence Handle1:CAS:528:DyaK3sXisFSnuro%3D Occurrence Handle10.1007/BF00587569

    Article  PubMed  CAS  Google Scholar 

  • T Longhurst E Lee R Hinde C Brady J Speirs (1994) ArticleTitleStructure of the tomato Adh2 gene and Adh2 pseudogenes, and a study of Adh2 gene expression in fruit Plant Mol Biol 26 1073–1084 Occurrence Handle7811967 Occurrence Handle1:CAS:528:DyaK2MXjtVWrsbc%3D Occurrence Handle10.1007/BF00040690

    Article  PubMed  CAS  Google Scholar 

  • DP Matton P Constable N Brisson (1990) ArticleTitleAlcohol dehydrogenase gene expression in potato following elicitor and stress treatment Plant Mol Biol 14 775–783 Occurrence Handle2102855 Occurrence Handle1:CAS:528:DyaK3cXkvFCqsLY%3D Occurrence Handle10.1007/BF00016510

    Article  PubMed  CAS  Google Scholar 

  • I Molina C Salles M Nicolas J Crouzet (1987) ArticleTitleGrape alcohol dehydrogenase. II Kinetics studies: mechanism, substrate and coenzyme specificity Am J Enol Vitic 38 60–64 Occurrence Handle1:CAS:528:DyaL2sXktVehsr4%3D

    CAS  Google Scholar 

  • S Morrell H Greenway (1989) ArticleTitleEvidence does not support ethylene as a cue for synthesis of alcohol dehydrogenase and pyruvate decarboxylase during exposure to hypoxia Austr J Plant Physiol 16 469–475 Occurrence Handle1:CAS:528:DyaK3cXntl2ltA%3D%3D Occurrence Handle10.1071/PP9890469

    Article  CAS  Google Scholar 

  • E Moyano S Encinas-Villarejo JA Lopez-Raez J Redondo-Nevado R Blanco-Portales ML Betillo C Sanz JL Caballero J Munoz-Blanco (2004) ArticleTitleComparative study between two strawberry pyruvate decarboxylase genes along fruit development and ripening, post-harvest and stress conditions Plant Sci 166 835–845 Occurrence Handle1:CAS:528:DC%2BD2cXhs12msLw%3D Occurrence Handle10.1016/j.plantsci.2003.09.025

    Article  CAS  Google Scholar 

  • K Nakajima T Hashimoto Y Yamada (1993) ArticleTitleTwo tropinone reductases with different stereospecificities are short-chain dehydrogenases evolved from a common ancestor Proc Natl Acad Sci USA 90 9591–9595 Occurrence Handle8415746 Occurrence Handle1:CAS:528:DyaK2cXnt1aruw%3D%3D Occurrence Handle10.1073/pnas.90.20.9591

    Article  PubMed  CAS  Google Scholar 

  • Nicholas KB, Nicholas HBJ (1997) GENEDOC: a tool for editing and annotating multiple sequence alignments, distributed by the authors

  • H-P Peng C-S Chan M-C Shih S-F Yang (2001) ArticleTitleSignalling events in the hypoxic induction of alcohol dehydrogenase gene in Arabidopsis Plant Physiol 126 742–749 Occurrence Handle11402202 Occurrence Handle1:CAS:528:DC%2BD3MXks1Gms70%3D Occurrence Handle10.1104/pp.126.2.742

    Article  PubMed  CAS  Google Scholar 

  • B Persson M Krook H Jörnvall (1991) ArticleTitleCharacteristics of short-chain alcohol dehydrogenases and related enzymes Eur J Biochem 200 537–543 Occurrence Handle1889416 Occurrence Handle1:CAS:528:DyaK3MXkvVyht7w%3D Occurrence Handle10.1111/j.1432-1033.1991.tb16215.x

    Article  PubMed  CAS  Google Scholar 

  • JS Peters C Frenkel (2004) ArticleTitleRelationship between alcohol dehydrogenase activity and low-temperature in two maize genotypes, Silverado F1 and Adh1–Adh2-doubly null Plant Physiol Biochem 42 841–846 Occurrence Handle15596104 Occurrence Handle1:CAS:528:DC%2BD2cXhtVOitbbM Occurrence Handle10.1016/j.plaphy.2004.10.004

    Article  PubMed  CAS  Google Scholar 

  • S Picton J Gray S Barton U AbuBakar A Lowe D Grierson (1993) ArticleTitlecDNA cloning and characterisation of novel ripening-related mRNAs with altered patterns of accumulation in the ripening inhibitor (rin) tomato ripening mutant Plant Mol Biol 23 193–207 Occurrence Handle8219049 Occurrence Handle1:CAS:528:DyaK2cXitVWm Occurrence Handle10.1007/BF00021431

    Article  PubMed  CAS  Google Scholar 

  • N Saitou M Nei (1987) ArticleTitleThe neighbor-joining method: a new method for reconstructing phylogenetic trees Mol Biol Evol 4 406–425 Occurrence Handle3447015 Occurrence Handle1:STN:280:DyaL1c7ovFSjsA%3D%3D

    PubMed  CAS  Google Scholar 

  • JJ Salas J Sanchez (1998) ArticleTitleAlcohol dehydrogenases from olive (Olea europaea) fruit Phytochemistry 48 35–40 Occurrence Handle9621451 Occurrence Handle1:CAS:528:DyaK1cXjsVymtro%3D Occurrence Handle10.1016/S0031-9422(97)01097-2

    Article  PubMed  CAS  Google Scholar 

  • P Sarni-Manchado C Verriès C Tesnière (1997) ArticleTitleMolecular characterization and structural analysis of one alcohol dehydrogenase gene (GV-Adh1) expressed during ripening of grapevine (Vitis vinifera L.) berry Plant Sci 125 177–187 Occurrence Handle1:CAS:528:DyaK2sXktF2gtb0%3D Occurrence Handle10.1016/S0168-9452(97)04630-X

    Article  CAS  Google Scholar 

  • J Speirs E Lee K Holt YD Kim NS Scott B Loveys W Schuch (1998) ArticleTitleGenetic manipulation of alcohol deshydrogenase levels in ripening tomato fruit affects the balance of some flavour aldehydes and alcohols Plant Physiol 117 1047–1058 Occurrence Handle9662548 Occurrence Handle1:CAS:528:DyaK1cXkvVyru78%3D Occurrence Handle10.1104/pp.117.3.1047

    Article  PubMed  CAS  Google Scholar 

  • J Speirs R Correll P Cain (2002) ArticleTitleRelationship between ADH activity, ripeness and softness in six tomato cultivars Sci Hort 93 137–142 Occurrence Handle1:CAS:528:DC%2BD38XhtlSlsbc%3D Occurrence Handle10.1016/S0304-4238(01)00316-8

    Article  CAS  Google Scholar 

  • C Tesnière M Pradal A El-Kereamy L Torregrosa P Chatelet JP Roustan C Chervin (2004) ArticleTitleInvolvement of ethylene signalling in a non-climacteric fruit: new elements regarding the regulation of ADH expression in grapevine J Exp Bot 55 2235–2240 Occurrence Handle15333642 Occurrence Handle10.1093/jxb/erh244

    Article  PubMed  Google Scholar 

  • C Tesnière C Verriès (2000) ArticleTitleMolecular cloning and expression of cDNAs encoding alcohol dehydrogenases from Vitis vinifera L. during berry development Plant Sci 157 77–88 Occurrence Handle10940471 Occurrence Handle10.1016/S0168-9452(00)00274-0

    Article  PubMed  Google Scholar 

  • JD Thompson TJ Gibson F Plewniak F Jeanmougin DG Higgins (1997) ArticleTitleThe ClustalX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools Nucleic Acids Res 24 4876–4882 Occurrence Handle10.1093/nar/25.24.4876

    Article  Google Scholar 

  • D Straeten ParticleVan der RA Rodrigues Pousada J Gielen M Montagu ParticleVan (1991) ArticleTitleTomato alcohol dehydrogenase. Expression during fruit ripening and under hypoxic conditions FEBS Lett 295 39–42 Occurrence Handle10.1016/0014-5793(91)81379-M

    Article  Google Scholar 

  • C Verriès M Pradal P Chatelet L Torregrosa C Tesnière (2004) ArticleTitleIsolation and analysis of the promoter of VvAdh2, a grapevine (Vitis vinifera L.) ripening-related gene Plant Sci 167 1067–1074 Occurrence Handle10.1016/j.plantsci.2004.06.003 Occurrence Handle1:CAS:528:DC%2BD2cXmsVGis7Y%3D

    Article  CAS  Google Scholar 

  • F Yahyaoui C Wongs-Aree A Latché R Hackett D Grierson JC Pech (2002) ArticleTitleMolecular and biochemical characteristics of a gene encoding an alcohol acyl-transferase involved in the generation of aroma volatile esters during melon ripening Eur J Biochem 269 2359–2366 Occurrence Handle11985619 Occurrence Handle1:CAS:528:DC%2BD38XjvFGjtrc%3D Occurrence Handle10.1046/j.1432-1033.2002.02892.x

    Article  PubMed  CAS  Google Scholar 

  • S Yokoyama DE Harry (1993) ArticleTitleMolecular phylogeny and evolutionary rates of alcohol dehydrogenases in vertebrates and plants Mol Biol Evol 10 1215–1226 Occurrence Handle8277852 Occurrence Handle1:CAS:528:DyaK2cXkvVCrtw%3D%3D

    PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. UMR 990 INRA/INPT-ENSAT “Génomique et Biotechnologie des Fruits”, Av. de l’Agrobiopole, BP 32607, F-31326 , Castanet-Tolosan Cedex, France

    Daniel Manríquez, Islam El-Sharkawy, Francisco B. Flores, Fikri El-Yahyaoui, Farid Regad, Mondher Bouzayen, Alain Latché & Jean-Claude Pech

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  1. Daniel Manríquez
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  2. Islam El-Sharkawy
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  3. Francisco B. Flores
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  4. Fikri El-Yahyaoui
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  5. Farid Regad
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  6. Mondher Bouzayen
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  7. Alain Latché
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  8. Jean-Claude Pech
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Corresponding author

Correspondence to Jean-Claude Pech.

Additional information

Daniel Manríquez and Islam El-Sharkawy contributed equally to the work. Accession numbers for Cm-ADH1 (ABC02081), and Cm-ADH2 (ABC02082).

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Manríquez, D., El-Sharkawy, I., Flores, F.B. et al. Two highly divergent alcohol dehydrogenases of melon exhibit fruit ripening-specific expression and distinct biochemical characteristics. Plant Mol Biol 61, 675–685 (2006). https://doi.org/10.1007/s11103-006-0040-9

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