Short communication Evidence of a sirtuin gene family in grapevine (Vitis vinifera L.) Matteo Busconi a , Serena Reggi c , Corrado Fogher a , Luigi Bavaresco b, * a Istituto di Botanica e Genetica vegetale, Universita` Cattolica del Sacro Cuore, Via Emilia Parmense, 84 29100 Piacenza, Italy b Istituto di Frutti-Viticoltura, Universita` Cattolica del Sacro Cuore, Via Emilia Parmense, 84 29100 Piacenza, Italy c Plantechno s.r.l., Via Staffolo, 60 26041 Vicomoscano, Cremona, Italy article info Article history: Received 5 November 2008 Accepted 10 March 2009 Available online 18 March 2009 Keywords: Sir2-like genes Vitis vinifera L. Genome Plants abstract Sirtuins are known as regulators of age-dependent gene transcription and chromatin modification in yeast and in animals, but information about their occurrence and role in plants is scarce. Sirtuin-like sequences were amplified using two highly degenerate primers designed comparing sirtuin sequences of seven different plant species, and characterised at the sequence level in Vitis vinifera L. cv. Barbera clone R4. The data were confirmed and analysed by searching the published database from two independent grape genome projects with the obtained sequences. Two putative sirtuin genes containing sirtuin- conserved domains were found on chromosome 7 and on chromosome 19, suggesting the occurrence of sirtuins in grapevine. Ó 2009 Elsevier Masson SAS. All rights reserved. 1. Introduction The sirtuin/Sir2 (Silent information regulator 2) family of NAD þ - dependent deacetylases and mono-ADP-ribosyltransferases are found in both prokaryotes and eukaryotes. Several sirtuins have been characterised in organisms [9], such as bacteria (S. solfataricus, L. major), yeast (S. cerevisiae), worms (C. elegans), fruit fly (D. mel- anogaster), and humans (H. sapiens). The name Sir2 is used for the S. cerevisiae, fruit fly and the roundworm sirtuin-type enzyme, while in mammals sirtuins are named SIRT1, SIRT2, SIRT3 etc., and SIRT1 is the mammalian ortholog of Sir2 [4]. In eukaryotes sirtuins affect cellular metabolism being involved in the regulation of transcriptional repression, recombination, the cell division cycle, and microtubule organisation [12] and it is claimed that they mediate the calorie restriction effect, extending lifespan. This latter aspect was investigated in yeast (S. cerevisiae), whose lifespan was extended by extra copies of Sir2 [8,10]. Overexpression of Sir2 homologs also extended the longevity of C. elegans [14] and D. melanogaster [13]. When yeast is deprived of food (calorie restriction) a stress pathway is activated, leading to an altered nicotinamide concentration and/or NAD:NADH ratio, which stim- ulates the activity of Sir2 [1,11]. The first plant sirtuin gene whose function was studied was a rice (Oryza sativa) Sir2-like gene. The role of this gene was investigated by Huang et al. [6], who found that is was required for safeguarding against genome instability and cell damage to ensure cell growth in rice. Sir2 occurrence in grapevine has not yet been assessed, even though the grapevine genome has recently been released [7,15]. The presence of Sir2-like sequences in the grape genome is intriguing, because stressed grapevine cells synthesize compounds with hydroxylated trans-stilbene ring structure, such as, resveratrol [2,5]. Resveratrol (trans 3,3 0 ,5-trihydroxystilbene) is able to activate sirtuins in yeast [5], in C. elegans and D. mela- nogaster [16], mimicking calorie restriction and extending lifespan. In this work we demonstrate, for the first time, the presence of Sir2 genes in grapevine. 2. Results and discussion When the experiment started only partial information con- cerning the genome sequences was available and database searches gave no results. Only a few data concerning plant sirtuins or sirtuin- like sequences were recovered (see Material and methods). With the exception of the record with the accession number AC189542, which refers to a genomic sequence showing introns and exons of Brassica rapa, all the remaining records were cDNA clones lacking introns. By aligning all the sirtuin-like plant sequences recovered in the database, we were able to design degenerate forward and reverse primers that were potentially specific for this gene family. Using these primers we successfully amplified DNA from grape, wheat, rice, maize, sorghum, arabidopsis, tomato, lettuce, cauli- flower, garlic, and alyssum (Fig. 1) but not from carrot, onion, basil, chicory, or olive even after modifying the amplification conditions. The negative results in the second group of species could be explained by differences in the annealing sites sequences in the * Corresponding author. Tel.: þ39 0523 599270; fax: þ39 0523 599268 . E-mail address: luigi.bavaresco@unicatt.it (L. Bavaresco). Contents lists available at ScienceDirect Plant Physiology and Biochemistry journal homepage: www.elsevier.com/locate/plaphy 0981-9428/$ – see front matter Ó 2009 Elsevier Masson SAS. All rights reserved. doi:10.1016/j.plaphy.2009.03.003 Plant Physiology and Biochemistry 47 (2009) 650–652