Fig 1.
Phylogenetic analysis of grape and Arabidopsis thaliana VPE family proteins.
Fig 2.
Intron and exon organization of Vitis vinifera VPE genes.
Table 1.
Gene analysis of VPEs in Vitis vinifera.
Fig 3.
Multi-sequence alignment of Vitis vinifera and Arabidopsis thaliana VPE proteins.
Amino acid sequences from 6 grapevine and 4 Arabidopsis thaliana VPE genes share similar catalytic dyad His (177), Cys (219) and the substrate binding pocket consisting of Arg (112), Arg (389), and Ser (395), with the exception of Ser (395) in VvγVPE, which was replaced with Ala (395).
Fig 4.
Expression of VPE genes in different grapevine tissues.
Expression of VPE genes analyzed by sqRT-PCR in root, stem, leaf, tendril, alabastrum, flowers, pericarp, pulp, and ovule tissues of Vitis vinifera cv. Pinot Noir.
Fig 5.
Expression of VvγVPE gene in different development stages of ovule in seed and seedless grapes.
Relative expression level of VvγVPE gene in different development stages (15, 20, 25, 30, 35, 40, and 45 DAF) of ovule in ‘V. vinifera cv. Thompson Seedless ‘, ‘V. vinifera cv. Youngle’, ‘V. vinifera cv. Pinot Noir’ and ‘V. vinifera cv.Flame Seedless’. DAF (days after full-bloom) (error bars indicate ±SD).
Fig 6.
Expression of VvδVPE gene in different development stages of ovule in seed and seedless grapes.
Relative expression level of VvδVPE gene in different development stages (15, 20, 25, 30, 35, 40, and 45 DAF) of ovule in ‘V. vinifera cv. Thompson Seedless ‘, ‘V. vinifera cv. Youngle’, ‘V. vinifera cv. Pinot Noir’ and ‘V. vinifera cv.Flame Seedless’. DAF (days after full-bloom) (error bars indicate ±SD).
Fig 7.
Expression of VvβVPE gene in different development stages of ovule in seed and seedless grapes.
Relative expression level of VvβVPE gene in different development stages (15, 20, 25, 30, 35, 40, and 45 DAF) of ovule in ‘V. vinifera cv. Thompson Seedless ‘, ‘V. vinifera cv. Youngle’, ‘V. vinifera cv. Pinot Noir’ and ‘V. vinifera cv.Flame Seedless’. DAF (days after full-bloom) (error bars indicate ±SD).
Fig 8.
Enzymatic activity of grapevine VPEs.
CK-1 (ddH2O); CK-2 (ddH2O and fluorescent VPE-specific substrate); GS115 (ddH2O, fluorescent VPE-specific substrate and GS115 thallus); pPICZαA (ddH2O, fluorescent VPE-specific substrate and GS115 thallus tansformed with pPICZαA plasmid); βVPE, γVPE and δVPE (ddH2O, fluorescent VPE-specific substrate and GS115 thallus transformed with recombinant plasmid). Enzymatic activity of recombinant grapevine βVPE, γVPE and δVPE were testified by using Ac-ESEN-MCA substrates at pH 5.5. Y-axes are scales of relative fluorescence values (error bars indicate ±SD).