Table 1.
Primer sequences used for qRT-PCR.
Table 2.
Primers used to generate template DNA for synthesize of RNA probes.
Figure 1.
Global CHI4D and TL3 mRNAs/proteins expression in berry during development.
A) Transcript accumulation of CHI4D and TL3 genes in Pinot noir grapevine berries during development. Analyses were performed by quantitative RT-PCR. Levels of transcripts were calculated using the standard curve method from duplicate data, with grapevine EF1-α gene as internal control and were expressed as mRNA copy number/1000 EF1-α mRNAs. Values represent the mean ± SD of duplicates of the harvest 2009. Letters indicate significant differences between BBCH stages as calculated by Mann-Whitney U test (P<0,05). The same trend was observed for 2008 and 2010. B) Western blot analysis of CHI4D and TL3 in Pinot noir berries during maturation. Results represent the harvest of 2009 the same trend was observed for 2008 and 2010.
Figure 2.
Localization of CHI4D and TL3 mRNAs expression sites in berry tissues during development.
In situ localization of CHI4D A) and TL3 B) mRNAs in Pinot noir berry tissues at BBCH75 (1, 2, 3, 4) and BBCH89 (5, 6, 7, 8) stages. Blue signal indicates the location of antisense (1, 2, 3, 5, 6, 7) and control sense (4, 8) probes. Sense probe controls indicate background staining. 1 and 5: central vascular bundles (CVB) in the mesocarp (MS), 2 and 6: peripheral vascular bundles (PVB) in the mesocarp, 3, 4, 7 and 8: exocarp (EX).
Figure 3.
Localization of CHI4D and TL3 proteins expression sites in berry tissues during development.
Immunolocalization of CHI4D A) and TL3 B) proteins in Pinot noir berry tissues at BBCH75 (1, 2, 3) and BBCH89 (4, 5, 6) stages. Controls without primary antibody indicate tissue background autofluorescence in berry tissues at BBCH75 (7) and BBCH89 (8) stages. 1 and 4: central vascular bundles (CVB) in the mesocarp (MS), 2 and 5: peripheral vascular bundles (PVB) in the mesocarp, 3, 6, 7 and 8: exocarp (EX). Epicuticular tissue (EP).
Figure 4.
Global CHI4D and TL3 mRNAs/proteins expression in UV-C treated berries. A)
Transcript accumulation of CHI4D (1, 2) and TL3 (3, 4) genes in Pinot noir grapevine berries 48 h after UV-C stress. Analyses were performed by quantitative RT-PCR. Levels of transcripts were calculated using the standard curve method from duplicate data, with grapevine EF1-α gene as internal control and were expressed as mRNA copy number/1000 EF1-α mRNA. Values represent the mean ± SD of duplicates of the harvest of 2009. Letters indicate significant differences between treatments at each BBCH stage as calculated by Mann-Whitney U test (P<0,05). The same trend was observed for 2008 and 2010. Abbreviations: untreated berries (NT), control detached berries (C), treated berries (T). B) Western blot analysis of CHI4D (1, 2) and TL3 (3, 4) in Pinot noir berries 48 h after an UV-C stress. Results represent the harvest of 2009. The same trend was observed for 2008 and 2010. Abbreviations: untreated berries (NT), control detached berries (C), treated berries (T).
Figure 5.
Localization of CHI4D and TL3 mRNAs expression sites in UV-C treated berries.
In situ localization of CHI4D A) and TL3 B) mRNAs in Pinot noir berry tissues at BBCH75 stages 48 h after UV-C irradiation. Blue signal indicates the location of antisense probes. 1: central vascular bundles (CVB) in the mesocarp (MS), 2: peripheral vascular bundles (PVB) in the mesocarp, 3: exocarp (EX).
Figure 6.
Localization of CHI4D and TL3 protein expression sites in UV-C treated berries.
Immunolocalization of CHI4D A) and TL3 B) proteins in Pinot noir berry tissues at BBCH75 stages 48 h after an UV-C irradiation. Green fluorescence indicates the localization of proteins. 1: central vascular bundles (CVB) in the mesocarp (MS), 2: peripheral vascular bundles (PVB) in the mesocarp, 3: exocarp (EX). Epicuticular tissue (EP).