Neutral invertases in grapevine and comparative analysis with Arabidopsis, poplar and rice.
Neutral invertases (NIs, EC 3.2.1.26) cleave sucrose to glucose and fructose. They are encoded by a small gene family of 9 members in the Arabidopsis genome, 8 in rice, 16 in poplar and 9 in Vitis vinifera (L.). The grapevine NIs were identified in the 8.4X genome assembly of the quasi-homozygous line PN40024. In addition, alleles of three NIs were sequenced in the heterozygous cultivar 'Cabernet Sauvignon'. Analyses of sequence variation between alleles, homoeologous and paralogous copies in grapevine and their orthologues in Arabidopsis, poplar and rice are provided. In grapevine, NIs were classified into four alpha NIs and five beta NIs and subsequently grouped into hierarchical clades using a combination of evidence including amino acid identity, exon/intron structure, rate of synonymous substitutions (K (s)) and chromosomal distribution. Estimation of K (s) proved the ancient origin of all NIs and the lack of expansion by gene duplication past the event of polyploidisation. We then focused on transcription analysis of five NIs for which evidence of expression was available from expressed sequence tag databases. Among these, four NIs consisted of pairs of homoeologous copies, each pair lying on a pair of chromosomes duplicated by polyploidy. Unequal expression of homoeologous genes was observed by quantitative RT-PCR in leaf, flower, seed and root tissues. Since NIs might play significant roles in fruit and wine quality, NIs expression was monitored in flesh and skin of 'Merlot' berries and shown in parallel with the suite of changes that accompany fruit ripening, including glucose and fructose accumulation.
Nonis
Alberto
A
Dipartimento di Scienze Agrarie e Ambientali, University of Udine, via delle Scienze 208, 33100 Udine, Italy.
Ruperti
Benedetto
B
Pierasco
Alessandro
A
Canaguier
Aurelie
A
Adam-Blondon
Anne-Françoise
AF
Di Gaspero
Gabriele
G
Vizzotto
Giannina
G
D000483:Alleles; D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics; D003062:Codon / Q000235:genetics; D019143:Evolution, Molecular; D005632:Fructose / Q000378:metabolism; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant; D018745:Genome, Plant / Q000235:genetics; D005947:Glucose / Q000378:metabolism; D006579:Heterozygote; D005810:Multigene Family; D009154:Mutation / Q000235:genetics; D012275:Oryza / Q000201:enzymology* / Q000235:genetics; D010802:Phylogeny; D032107:Populus / Q000201:enzymology* / Q000235:genetics; D021381:Protein Transport; D017422:Sequence Analysis, DNA; D027843:Vitis / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D043324:beta-Fructofuranosidase / Q000235:genetics* / Q000378:metabolism
D003062:Codon; D005632:Fructose; D043324:beta-Fructofuranosidase; D005947:Glucose
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