Journal of Experimental Botany , Vol. Page 63, 63, 1 ofNo. No. 16 11, 2, pp. pp. 695–709, 4275–4290, 2012 2012 doi:10.1093/jxb/err313doi:10.1093/jxb/ers120 AdvanceAdvance AccessAccess publicationpublication 44 November,May, 2012 2011 This paper is available online free of all access charges (see http://jxb.oxfordjournals.org/open_access.html for further details) RESEARCH PAPER Journal of Experimental Botany 63 InThePosidonia fruit ripening-related oceanica cadmium gene FaAAT2 inducesencodes changes an in acyl DNA 11 4275 methylationtransferase involved and chromatin in strawberry patterning aroma biogenesis 4290 2012 MariaGuadalupe Greco, Cumplido-Laso Adriana Chiappetta,1, Laura Leonardo Medina-Puche Bruno1, and Enriqueta Maria Beatrice Moyano1 Bitonti*, Thomas Hoffmann2, Quirin Sinz2, 2 2 1 2 1 doi:10.1093/jxb/ers120 DepartmentLudwig Ring of Ecology,, Claudia University Studart-Wittkowski of Calabria, Laboratory, Jose of´ PlantLuis Cyto-physiology, Caballero , Wilfried Ponte Pietro Schwab Bucci,,JuanMun I-87036 Arcavacata˜ oz-Blanco di Rende, Advance Access publication 4 May, 2012 Cosenza,and Rosario Italy Blanco-Portales1,* Downloaded from https://academic.oup.com/jxb/article/63/11/4275/603789 by guest on 30 September 2021 Cumplido-Laso et al. *1 ToDepartamento whom correspondence de Bioquı´mica should y Biologı be addressed.´a Molecular. E-mail: Edificio [email protected] C-6, Campus Universitario de Rabanales y Campus de Excelencia Strawberry FaAAT2 is involved in fruit aroma biogenesis Internacional Agroalimentario CEIA3, Universidad de Co´ rdoba, 14071 Co´ rdoba, Spain 2 ReceivedBiotechnology 29 May of2011; Natural Revised Products, 8 July 2011; Technische Accepted Universita 18 August¨ t Mu¨ 2011nchen, Liesel-Beckmann-Str. 1, 85354 Freising, Germany * To whom correspondence should be addressed. E-mail: [email protected] ReceivedAbstract 6 February 2012; Revised 19 March 2012; Accepted 26 March 2012 In mammals, cadmium is widely considered as a non-genotoxic carcinogen acting through a methylation-dependent epigenetic mechanism. Here, the effects of Cd treatment on the DNA methylation patten are examined together with itsAbstract effect on chromatin reconfiguration in Posidonia oceanica. DNA methylation level and pattern were analysed in Short-chainactively growing esters organs, contribute under toshort- the (6 blend h) and of long- volatiles (2 d or that 4 d) define term and the low strawberry (10 mM) aroma. and high The (50 lastmM) stepdoses in of their Cd, biosynthesisthrough a Methylation-Sensitive involves an alcohol acyltransferase Amplification that Polymorphism catalyses the technique esterification and of an an immunocytological acyl moiety of acyl-CoA approach, with anrespectively. alcohol. This The study expression identified of aone novel member strawberry of the alcoholCHROMOMETHYLASE acyltransferase gene(CMT (FaAAT2) family,) awhose DNA expressionmethyltransferase, pattern duringwas also fruit assessed receptacle by growth qRT-PCR. and ripeningNuclear ischromatin in accordance ultrastructure with the was production investigated of esters by throughout transmission strawberry electron fruitmicroscopy. ripening. Cd The treatment full-length inducedFaAAT2 a DNAcDNA hypermethylation, was cloned and as expressed well as an inup-regulationEscherichia of coliCMTand, indicating its activity that wasde analysednovo methylation with acyl-CoA did indeed and alcohol occur. substrates. Moreover, Thea high semi-purified dose of Cd FaAAT2 led to enzymea progressive had activity heterochromatinization with C1–C8 straight- of chaininterphase alcohols nuclei and and aromatic apoptotic alcohols figures in were the alsopresence observed of acetyl-CoA. after long-term Cinnamyl treatment. alcohol The was data the demonstrate most efficient that acyl Cd acceptor.perturbs the When DNAFaAAT2 methylationexpression status was through transiently the involvement downregulated of a specific in the fruit methyltransferase. receptacle by agroinfiltration, Such changes arethe volatilelinked to ester nuclear production chromatin was reconfiguration significantly reduced likely toin establish strawberry a new fruit. balance The results of expressed/repressed suggest that FaAAT2 chromatin.plays aOverall, significant the data role show in the an production epigenetic of basis esters to that the contribute mechanism to underlying the final strawberry Cd toxicity fruit in plants. flavour. Key words: Alcohol5-Methylcytosine-antibody, acyltransferase, BADH, cadmium-stress fruit aroma, strawberry, condition, chromatin volatile compounds. reconfiguration, CHROMOMETHYLASE, DNA-methylation, Methylation- Sensitive Amplification Polymorphism (MSAP), Posidonia oceanica (L.) Delile. Introduction Introduction Volatile esters play several roles in higher plants. They only about 20 of these compounds actually contribute to Inmake the flowers Mediterranean more attractive coastal to pollinators ecosystem, and the dispersing endemic itsAlthough aroma and not flavour essential (Forney for plantet al., growth, 2000). Some in terrestrial authors seagrassanimals,Posidonia act as protectants oceanica (L.) against Delile pathogens plays a relevant by inducing role plants,consider Cd methyl is readily butanoate, absorbed ethyl by roots butanoate, and translocated methyl hex- into byseveral ensuring important primary plant production, defence pathways, water andoxygenation contribute and to aerialanoate, organs hexyl while, acetate, in acquatic and ethyl plants, hexanoate it is directly in addition taken up to providesthe aroma niches of ripe for somefruit (D’Auria animals, besideset al.,counteracting 2007). These by2,5-dimethyl-4-hydroxy-3(2H)-furanone leaves. In plants, Cd absorption induces (HDMF, complex furaneol) changes coastalcompounds erosion are through produced its widespread by all soft meadows fruit species (Ott, during 1980; atand the its genetic, methyl ether biochemical as the mostand physiological important constituents levels which of Piazziripeninget and al., play 1999; an Alcoverro importantet role al., determining 2001). There the is final also ultimatelythe characteristic account strawberry for its toxicity aroma (Valle (Larsen andet al. Ulmer,, 1992; 1972; Jetti considerablesensory quality evidence of fruit that (ManrıP.´ oceanicaquez et al.plants, 2006). are In able fruits to Sanitet al., 2007).di Toppi Analysis and Gabrielli, of volatiles 1999; from Benavides fruitet of al. different, 2005; z absorblike apple and (Malus accumulate domestica metals), pear from (Pyrus sediments communis (Sanchiz), and WeberFragariaetvarieties al., 2006; has Liu identifiedet al., some 2008). common The most esters obvious (ethyl etbanana al., 1990; (Musa Pergent-Martini, sapientum), esters 1998; are Maserti the mainet al. components, 2005) thus symptomacetate, methyl of Cd toxicity butanoate, is a reduction 2-methylbutyl in plant acetate, growth due octyl to influencingof their characteristic metal bioavailability aroma, while in the in marine strawberry ecosystem. they anacetate, inhibition octyl of butanoate, photosynthesis, hexyl acetate, respiration, ethyl and heptanoate, nitrogen Foronly this contribute reason, to this the seagrass blend of is widelyvolatiles considered that define to the be metabolism,2-hexenyl butanoate, as well as benzyl a reduction acetate, in and water hexyl and 2-methyl mineral aaroma metal (Beekwilder bioindicatoret species al., 2004). (Maserti The strawberryet al., 1988; aroma Pergent is uptakebutanoate) (Ouzonidou in bothetFragaria al., 1997; chiloensis Perfus-Barbeochfruits andet al. several, 2000; etdetermined al., 1995; by Lafabrie moreet than al., 300 2007). compounds Cd is one consisting of most FragariaShukla et al.3, 2003;ananassa Sobkowiakcultivars and (Drawert Deckert, 2003).et al., 1973; widespreadof alcohols, heavy aldehydes, metals esters, in both sulphur terrestrial compounds, and marine and HirviAt theand genetic Honkanen, level, 1982). in both However, animals some and esters plants, found Cd environments.furanone derivatives (Zabetakis and Holden, 1997), but incanF. induce chiloensis chromosomal(hexyl propanoate, aberrations, ethyl abnormalities 4-decenoate, in ª© 2011The Author The Author(s). [2012]. Published by Oxford University Press [on behalf of the the Society Society for for Experimental Experimental Biology]. Biology]. All All rights rights reserved. reserved. ThisFor Permissions, is an Open Access please articlee-mail: distributed [email protected] under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by- nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. 42762 of 16 | Cumplido-Laso| Cumplido-Laso et etal. al. 2-phenylethyl propanoate, and ethyl 2,4-decadienoate) have red-ripened fruits, suggesting that their corresponding not been described previously in F. 3 ananassa (Pyysalo enzymes are involved in the biosynthesis of volatile esters et al., 1979; Zabetakis et al., 1997; Azodanlou et al., 2003; in the strawberry fruit receptacle. The SAAT enzyme Berna et al., 2007; Gonza´lez et al., 2009). prefers medium-chain aliphatic alcohols in combination The last step in the biosynthesis of volatile esters is with different acyl-CoAs as substrates. In contrast, with catalysed by alcohol acyltransferase (AAT), a key enzyme in the tertiary monoterpenol linalool as substrate, activity aroma biochemistry (Fellman et al., 2000). This enzyme was insignificant. Although the