Figure captions
Figure 1. Amounts of sugars quantified for each ripening stage. Experiments were performed with strawberry fruits immediately after harvest at ripening stages AR: 3/4 red, FR: full red and DR: dark red. A) Sucrose (mg g-1 FW); B) Glucose (mg g-1 FW); C) Fructose (mg g-1 FW); D) Myo-inositol (µg g-1 FW); E) Trehalose (µg g-1 FW); F) 1-Kestose (µg g-1 FW); G) 6G-Kestose (µg g-1 FW); H) Nystose (µg g-1 FW); I) Kestopentaose (µg g-1FW); J) Raffinose (µg g-1 FW); K) Estaquiose (µg g-1 FW)). Graphs represent the average of at least three independent replicates (each one contains 10 fruits) quantified. Error bars indicate ± SEM; ***P < 0.001, ****P< 0.0001; Student’s t‐test of AR versus FR or DR value.
Figure 2 . Phylogenetic analysis of main sucrose metabolism proteins. Protein sequence phylogeny using the Maximum Likelihood method and JTT matrix-based model for Fragaria vesca andArabidopsis thaliana in: A) SPS, B) SS, C) CWINV and VINV proteins families. HMM profiles to each protein family from Pfam or InterPro databases were used as a query against the F. vescav4.0.a2 genome to find the closest F. vesca homologues. These paralogues were used for find the most closed Arabidopsisorthologues.
Figure 3. Expression pattern of the main sucrose metabolism genes during different ripening stages. Experiments were performed with strawberry fruits immediately after harvest at ripening stages AR: 3/4 red, FR: full red and DR: dark red. Expression levels of: A)FvSPS1 and FvSPS2 , B) FvSS1 and FvSS2 , C)FvCWINV1 and FvCWINV2 and D) FvVINV1 andFvVINV2 . E) Pictures of strawberry fruits at different ripening stages. Graphs represent the average of three independent biological experiments quantified by qPCR. Error bars indicate ± SEM; *P< 0.05, **P < 0.01, ***P < 0.001; Student’s t‐test of AR versus FR or DR value.
Figure 4. Amounts of sugars quantified for each day during storage at 0 ºC. Experiments were performed with strawberry fruit at 0d: immediately after harvest; 2d Air, 7d Air: air stored fruits during 2 or 7 days; 2d CO2: fruits treated during 2 days with CO2; 2d CO2+5d Air: fruits treated during 2 days with CO2 plus 5 days in air. A) Sucrose (mg g-1 FW); B) Glucose (mg g-1 FW); C) Fructose (mg g-1 FW); D) Myo-inositol (µg g-1 FW); E) Trehalose (µg g-1 FW); F) 1-Kestose (µg g-1 FW); G) 6G-Kestose (µg g-1 FW); H) Nystose (µg g-1 FW); I) Kestopentaose (µg g-1 FW); J) Raffinose (µg g-1 FW); K) Estaquiose (µg g-1 FW)). Graphs represent the average of at least three independent replicates (each one contains 10 fruits) quantified. Error bars indicate ± SEM; *P < 0.05, **P < 0.01, ***P< 0.001, ****P < 0.0001; Student’s t‐test of 0d versus each treatment value.
Figure 5. Expression levels of the main sucrose metabolism genes during storage at 0ºC. Experiments were performed with strawberry fruit at 0d: immediately after harvest; 2d Air, 7d Air: air stored fruits during 2 or 7 days; 2d CO2: fruits treated during 2 days with CO2; 2d CO2+5d Air: fruits treated during 2 days with CO2 plus 5 days in air. Expression levels of: A) FvSPS1 and FvSPS2 , B)FvSS1 and FvSS2 , C) FvCWINV1 and FvCWINV2and D) FvVINV1 and FvVINV2 . Graphs represent the average of three independent biological experiments quantified by qPCR. Error bars indicate ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; Student’s t‐test of 0d versus each treatment value.
Figure 6. Amounts of sugars quantified during shelf-life at 20 ºC. Experiments were performed with SLC: freshly harvested fruit placed directly at 20 ºC during two days; LTA: samples transferred 1 day at 20 ºC from air at low temperature; LTC: samples transferred 1 day at 20 ºC from CO2 pretreatment. A) Sucrose (mg g-1 FW); B) Glucose (mg g-1 FW); C) Fructose (mg g-1 FW); D) Myo-inositol (µg g-1 FW); E) Trehalose (µg g-1 FW); F) 1-Kestose (µg g-1 FW); G) 6G-Kestose (µg g-1 FW); H) Nystose (µg g-1 FW); I) Kestopentaose (µg g-1 FW); J) Raffinose (µg g-1 FW); K) Estaquiose (µg g-1 FW). Graphs represent the average of at least three independent replicates (each one contains 10 fruits) quantified. Error bars indicate ± SEM; *P < 0.05, **P < 0.01, ***P< 0.001; Student’s t‐test of SLC versus LTA or LTC value.
Figure 7. Expression levels of the main sucrose metabolism genes during shelf-life at 20 ºC. Experiments were performed with SLC: freshly harvested fruit placed directly at 20 ºC during two days; LTA: samples transferred 1 day at 20 ºC from air at low temperature; LTC: samples transferred 1 day at 20 ºC from CO2pretreatment. A) Sucrose (mg g-1 FW); B) Glucose (mg g-1 FW); C) Fructose (mg g-1 FW); D) Myo-inositol (µg g-1 FW); E) Trehalose (µg g-1 FW). Graphs represent the average of three independent biological experiments quantified by qPCR. Error bars indicate ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Student’s t‐test of SLC versus LTA or LTC value.
Figure 8. Changes in weight loss (%) during storage and during shelf-life compared to strawberry fruit immediately after harvest. Fruit weight losses were measured in strawberry fruit at 2d Air, 7d Air: air stored fruits during 2 or 7 days; 2d CO2: fruits treated during 2 days with CO;, 2d CO2+5d Air: fruits treated during 2 days with CO2 plus 5 days in air; SLC: freshly harvested fruit placed directly at 20 ºC during two days; LTA: samples transferred 1 day at 20 ºC from air at low temperature; LTC: samples transferred 1 day at 20 ºC from CO2 pretreatment. Graphs represent the average of at least 16 (n=16-52) independent experiments. Error bars indicate ± SEM; **P < 0.01, ****P < 0.0001. Student’s t‐test of each comparison is indicated.