Abstract
Flavour is an important key factor of apple (Malus × domestica Borkh.) fruit quality, and its improvement is an important but complex breeding goal. Acetate esters are quantitatively the most important volatile compounds in apple fruit, and only a few of them dominate the typical aroma of a cultivar. Alcohol acyl-transferase (AAT) is a key enzyme involved in the last step of ester biosynthesis. The aim of this study was to target single nucleotide polymorphisms (SNPs) in an AAT candidate gene genetically associated with ester quantitative trait loci (QTL), to enable functional marker development for marker-assisted apple breeding programs. The AAT gene inventory of apple was characterized by in-silico mining of the assembled Golden Delicious genome, and 17 putative AAT genes in total were defined. MdAAT1 located on chromosome 2 was selected as the main candidate gene associated with QTL for different acetate esters, and its allelic diversity was assessed by direct amplicon sequencing in a collection of 102 apple cultivars characterized for ester volatile profiles. Sequencing a 468 bp nucleotide sequence of the MdAAT1 coding region resulted in the detection of four SNPs. In total, 18 different SNP haplotypes/heterozygous patterns were generated from the four SNPs identified within the apple collection. Association analyses resulted in highly significant associations of both individual SNPs and distinct haplotypes with the content of four acetate esters, including hexyl acetate, butyl acetate and 2-methyl-butyl acetate. About a third (31) of the 102 apple cultivars possessed the specific MdAAT1 haplotype H1 (C-A-C-A) and were characterized by strongly decreased ester concentrations. The contrasting H8 haplotype (T-G-T-G) was found in 28 varieties but was associated with normal to elevated ester concentrations. The observed association suggests a putative causal functional relationship between MdAAT1 and production of key apple esters.
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Aharoni A, Keizer LCP, Bouwmeester HJ, Sun Z, Alvarez-Huerta M, Harrie A, Verhoeven HA, Blaas J, van Houwelingen AMML, De Vos RCH, van der Voet H, Jansen RC, Guis M, Mol J, Davis RW, Schena M, van Tunen AJ, O’Connell AP (2000) Identification of the SAAT gene involved in strawberry flavor biogenesis by use of DNA microarrays. Plant Cell 12:647–661
Altisent R, Echeverria G, Graell J, Lopez L, Lara I (2009) Lipoxygenase activity is involved in the regeneration of volatile ester-synthesizing capacity after ultra-low oxygen storage of ‘Fuji’ apple. J Agric Food Chem 57:4305–4312
Andersen JR, Lübberstedt T (2003) Functional markers in plants. Trends Plant Sci 8:554–560
Balbontin C, Gaete-Eastman C, Fuentes L, Figueroa CR, Herrera R, Manriques D, Latche A, Pech JC, Moya-Leon MA (2010) VpAAT1: a gene encoding an alcohol acyltransferase, is involved in ester biosynthesis during ripening of montain papaya fruit. J Agric Food Chem 58:5114–5121
Beekwilder J, Alvarez-Huerta M, Neef E, Verstappen FWA, Bouwmeester HJ, Aharoni A (2004) Functional characterization of enzymes forming volatile esters from strawberry and banana. Plant Physiol 135:1865–1878
Bradbury PJ, Zhang Z, Kroon DE, Casstevens TM, Ramdoss Y, Buckler ES (2007) TASSEL: software for association mapping of complex traits in diverse samples. Bioinformatics 23:2633–2635
Costa F, Stella S, Van de Weg WE, Guerra W, Cecchinel M, Dallavia J, Koller B, Sansavini S (2005) Role of the genes Md-ACO1 and Md-ACS1 in ethylene production and shelf life of apple (M. domestica Borkh). Euphytica 141:181–190
Costa F, Van de Weg WE, Stella S, Dondini L, Pratesi D, Musacchi S, Sansavini S (2008) Map position and functional diversity of Md-Exp7: a new putative expansin gene associated with fruit softening in apple (Malus × domestica Borkh.) and pear (P. communis). Tree Genet Genomes 4:575–586
Costa F, Peace CP, Stella S, Serra S, Musacchi S, Bazzani M, Sansavini S, Van de Weg WE (2010) QTL dynamics for fruit firmness and softening around an ethylene-dependent polygalacturonase gene in apple (Malus x domestica Borkh.). J Exp Bot 61:3029–3039
D’Auria JC (2006) Acyltransferases in plants: a good time to be BAHD. Curr Opin Plant Biol 9:331–340
Defilippi BG, Dandekar AM, Kader AA (2005) Relationship of ethylene biosynthesis to volatile production, related enzymes, and precursor availability in apple peel and flesh tissues. J Agric Food Chem 53:3133–3141
Dixon J, Hewett EW (2000) Factors affecting apple aroma/flavour volatile concentration: a review. NZ J Crop Hort Sci 28:155–173
Dunemann F, Ulrich D, Boudichevskaia A, Grafe C, Weber WE (2009a) QTL mapping of aroma compounds analyzed by headspace solid-phase microextraction gas chromatography in the apple progeny Discovery × Prima. Mol Breed 23:501–521
Dunemann F, Boudichevskaia A, Grafe C, Weber WE, Ulrich D (2009b) QTL and candidate gene mapping for aroma compounds in the apple progeny Discovery × Prima. Acta Hort 839:433–440
Echeverria G, Graell J, Lopez ML, Lara I (2004) Volatile production, quality and aroma-related enzyme activities during maturation of ‘Fuji’ apples. Postharvest Biol Technol 31:217–227
El-Sharkawy I, Manriques D, Flores FB, Regad F, Bouzayen M, Latche A, Pech JC (2005) Functional 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 59:345–362
Evans KM, Patocchi A, Rezzonico F, Mathis F, Durel CE, Fernández-Fernández F, Boudichevskaia A, Dunemann F, Stankiewicz-Kosyl M, Gianfranceschi L, Komjanc M, Lateur M, Madduri M, Noordijk Y, Van de Weg WE (2010) Genotyping of pedigreed apple breeding material with a genome-covering set of SSRs: trueness–type of cultivars and their parentages. Mol Breed 56(7):361–378. doi:10.1007/s11032-010-9502-5
Fuhrmann E, Grosch W (2002) Character impact odorants of the apple cultivars Elstar and Cox Orange. Nahrung/Food 46:187–193
Ganal MW, Altmann T, Röder MS (2009) SNP identification in crop plants. Curr Opin Plant Biol 9:331–340
Knee M, Hatfield SGS (1981) The metabolism of alcohols by apple fruit tissue. J Sci Food Agric 32:593–900
Li D, Xu Y, Xu G, Gu L, Li D, Shu H (2006) Molecular cloning and expression of a gene encoding alcohol acyltransferase (MdAAT2) from apple (cv. golden delicious). Phytochemistry 67:658–667
Liebhard R, Gianfranceschi L, Koller B, Ryder CD, Tarchini R, van de Weg WE, Gessler C (2002) Development and characterization of 140 new microsatellites in apple (Malus × domestica Borkh.). Mol Breed 10:217–241
Malysheva-Otto LV, Röder MS (2006) Haplotype diversity in the endosperm specific β-amylase gene Bmy1 of cultivated barley (H. vulgare L.). Mol Breed 18:143–156
Matthies IE, Weise S, Förster J, Röder MS (2009) Association mapping and marker development of the candidate genes (1 → 3), (1 → 4)-β-D-Glucan-4-glucanohydrolase and (1 → 4)-β-Xylan-endohydrolase 1 for malting quality in barley. Euphytica 170:109–122
Noiton DAM, Alspach PA (1996) Founding clones, inbreeding, coancestry, and status number of modern apple cultivars. J Am Soc Hort Sci 121:773–782
Oraguzie NC, Iwanami H, Soejima J, Harada T, Hall A (2004) Inheritance of the Md-ACS1 gene and its relationship to fruit softening in apple (Malus × domestica Borkh.). Theor Appl Genet 108:1526–1533
Paillard NMM (1990) The flavour of apples, pears and quinces. In: Morton ID, Macleod AJ (eds) The flavour of fruits. Elsevier, Amsterdam, pp 1–41
Pritchard JK, Stephens M, Donnelly P (2000a) Inference of population structure using multilocus genotype data. Genetics 155:945–959
Pritchard JK, Stephens M, Rosenberg NA, Donnelly P (2000b) Association mapping in structured populations. Am J Hum Genet 67:170–181
Rafalski JA (2010) Association genetics in crop improvement. Curr Opin Plant Biol 13:174–180
Rowan DD, Lane HP, Allen JM, Fielder S, Hunt MB (1996) Biosynthesis of 2-methylbutyl, 2-methyl-2-butenyl and 2-methylbutanoate esters in red delicious and Granny Smith apples using deuterium-labeled substrates. J Agric Food Chem 44:3276–3285
Rowan DD, Allen JM, Fielder S, Hunt MB (1999) Biosynthesis of straight-chain ester volatiles in red delicious and Granny Smith apples using deuterium-labeled precursors. J Agric Food Chem 47:2553–2562
Rowan DD, Hunt MB, Dimouro A, Alspach PA, Weskett R, Volz RK, Gardiner SE, Chagné D (2009) Profiling fruit volatiles in the progeny of a Royal Gala × Granny Smith apple (Malus × domestica) cross. J Agric Food Chem 57:7953–7961
Saftner RA, Abbott JA, Bhagwat AA, Vinyard BT (2005) Quality measurement of intact and fresh-cut slices of Fuji, Granny Smith, pink lady and goldrush apples. J Food Sci 70:317–324
Saitou N, Nei M (1987) The neighbour-joining methods: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425
Schaffer RJ, Ellen N, Friel EN, Souleyre EJF, Bolitho K, Thodey K, Ledger S, Bowen JH, Ma J-H, Nain B, Cohen D, Gleave AP, Crowhurst RN, Janssen BJ, Yao J-L, Newcomb RD (2007) A genomics approach reveals that aroma production in apple is controlled by ethylene predominantly at the final step in each biosynthetic pathway. Plant Physiol 144:1899–1912
Schwab W, Davidovich-Rikanati R, Lewinsohn E (2008) Biosynthesis of plant-derived flavor compounds. Plant J 54:712–732
Silfverberg-Dilworth E, Matasci CL, van de Weg WE, van Kaauwen MPW, Walzer M, Kodde LP, Soglio V, Gianfranceschi L, Durel CE, Costa F, Yamamoto T, Koller B, Gessler C, Patocchi A (2006) Microsatellite markers spanning the apple (Malus × domestica Borkh.) genome. Tree Genet Genomes 2:202–224
Singh A, Singh PK, Singh R, Pandit A, Mahato AK, Gupta DK, Tyagi K, Singh AK, Singh NK, Sharma TR (2010) SNP haplotypes of the BADH1 gene and their association with aroma in rice (O. sativa L.). Mol Breed 26:325–338
Song J, Fourney CF (2008) Flavour volatile production and regulation in fruit. Can J Plant Sci 88:537–550
Souleyre EJF, Greenwood DR, Friel EN, Karunairetnam S, Newcomb RD (2005) An alcohol acyl transferase from apple (cv. Royal Gala), MpAAT1, produces esters involved in apple fruit flavor. FEBS J 272:3132–3144
Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA 4: molecular evolutionary genetics analysis software version 4.0. Mol Biol and Evol 24:1596–1599
Ulrich D, Hoberg E, Fischer C (2009) Diversity and dynamic of sensory related traits in different apple cultivars. J Appl Bot Food Qual 83:70–75
Velasco R, Zharkikh A, Affourtit J, Dhingra A, Cestaro A, Kalyanaraman A, Fontana P, Bhatnagar SK, Troggio M, Pruss D, Salvi S, Pindo M, Baldi P, Castelletti S, Cavaiuolo M, Coppola G, Costa F, Cova V, Dal Ri A, Goremykin V, Komjanc M, Longhi S, Magnago P, Malacarne G, Malnoy M, Micheletti D, Moretto M, Perazzolli M, Si-Ammour A, Vezzulli S, Zini E, Eldredge G, Fitzgerald LM, Gutin N, Lanchbury J, Macalma T, Mitchell JT, Reid J, Wardell B, Chen Z, Desany B, Niazi F, Palmer M, Koepke T, Jiwan D, Schaeffer S, Krishnan V, Wu C, Chu VT, King ST, Vick J, Tao Q, Mraz A, Stormo A, Stormo K, Bogden R, Ederle D, Stella A, Vecchietti A, Kater MM, Masiero S, Lasserre P, Lespinasse Y, Allan AC, Bus V, Chagné D, Crowhurst RN, Gleave AP, Lavezzo E, Fawcett JA, Proost S, Rouzé P, Sterck L, Toppo S, Lazzari B, Hellens RP, Durel CE, Gutin A, Bumgarner RE, Gardiner SE, Skolnick M, Egholm M, Van de Peer Y, Salamini F, Viola R (2010) The genome of the domesticated apple (Malus × domestica Borkh.). Nat Genet 42:833–839
Wang J, De Luca V (2005) The biosynthesis and regulation of biosynthesis of Concord grape fruit esters: including foxy methyl anthranilate. Plant J 44:606–619
Young JC, Chu CL, Lu X, Zhu H (2004) Ester variability in apple varieties as determined by solid-phase microextraction and gas chromatography-mass spectrometry. J Agric Food Chem 52:8086–8093
Zhu Y, Barritt BH (2008) Md-ACS1 and Md-ACO1 genotyping of apple (Malus × domestica Borkh.) breeding parents and suitability for marker-assisted selection. Tree Genet Genomes 4:555–562
Zhu Y, Rudell DR, Mattheis JP (2008) Characterization of cultivar differences in alcohol acyltransferase and 1-aminocyclopropane-1-carboxylate synthase gene expression and volatile ester emission during apple fruit maturation and ripening. Postharvest Biol Technol 49:330–339
Zini E, Biasioli F, Gasperi F, Mott D, Aprea E, Märk TD, Patocchi A, Gessler C (2005) QTL mapping of volatile compounds in ripe apples detected by proton transfer reaction-mass spectrometry. Euphytica 145:269–279
Acknowledgments
This research was funded in part through the European project ISAFRUIT (2006–2010). ISAFRUIT was funded by the European Commission under the Thematic Priority 5: Food Quality and Safety of the 6th Framework Program of RTD (Contract no. FP6-FOOD-CT-2006-016279). The authors are grateful to Regina Gläss, Astrid Sahre, Jürgen Egerer and Kirsten Weiß for their technical assistance. The authors also thank Alessandro Cestaro for bioinformatics support towards AAT gene mining and annotation.
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Dunemann, F., Ulrich, D., Malysheva-Otto, L. et al. Functional allelic diversity of the apple alcohol acyl-transferase gene MdAAT1 associated with fruit ester volatile contents in apple cultivars. Mol Breeding 29, 609–625 (2012). https://doi.org/10.1007/s11032-011-9577-7
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DOI: https://doi.org/10.1007/s11032-011-9577-7