G. Ferrara 1, A. Mazzeo 1, A.M.S. Matarrese 1, C. Pacucci 1, V. Gallo 2,3

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1 G. Ferrara 1, A. Mazzeo 1, A.M.S. Matarrese 1, C. Pacucci 1, V. Gallo 2,3 1 Dipartimento di Scienze del Suolo, della Pianta e degli Alimenti - University of Bari Aldo Moro, via Amendola 165/A Bari (Italy) 2 DICATECh, Politecnico di Bari, via Orabona Bari (Italy) 3 Innovative Solutions S.r.l. Spin Off company of Politecnico di Bari, zona H 150/B Noci (Bari, Italy)

2 INTRODUCTION In Italy, 41% of table grape area is cultivated with cultivar Italia with a yield of 574,000 tons (Puglia Region, 2010). Puglia, in Southeastern Italy, is the most important region for table grape production (32,450 ha) and Italia table grape (15,000 ha) is one of most important cultivars in the overall regional table grape production (ISMEA, 2012).

3 INTRODUCTION Berry size is the main quality factor affecting sales of table grapes in international markets. Berry thinning Crop load Plant growth regulators Trunk girdling

4 GIRDLING Removing a ring of bark from either the trunk or cane Restriction of movement of assimilates from aerial portion to roots Sap flow to buds, clusters. The interruption of the phloem is temporary. LARGER BERRY

5 Plant hormones Auxins, gibberellins cytokinins, ethylene, abscisic acid, jasmonic acid, etc. Plant growth regulators (PGRs) Natural origin Synthetic GIBBERELLIC ACID (GA 3 ) FORCHLORFENURON (CPPU)

6 GA 3 & table grape INCREASE OF RNA SYNTHESIS INCREASE OF PROETINS SYNTHESIS STIMULATION OF CELL ELONGATION AND DIVISION HIGHER SUGAR AND WATER INTAKE INTO THE CELLS Increase of weight and size of the berry

7 CPPU & table grape INCREASE OF RNA SYNTHESIS INCREASE OF PROETINS SYNTHESIS BERRY ENLARGMENT INTAKE OF ASSIMILATES FROM THE PHLOEM Increase of weight and size of the berry

8 GIRDLING GA 3 CPPU Morphological analyses Chemical analyses Metabolomic approach Yield Qualitative parameters

9 LOCATION: Commercial vineyard in Turi (Bari province) 2011 and 2012 CULTIVAR: ITALIA grafted onto 34 E.M Overhead trellis system Space: 2.4 m x 2.4 m Year of plantation: 2002 Vines/ha: 1740 TREATMENTS: T1: control T2: cane girdling T3: gibberellic acid, GA 3 (Berelex) T4: forchlorfenuron, CPPU (Sitofex) APPLICATION: motorized sprayer (volume 1000 L/ha)

10 PHENOLOGICAL PHASE AND TIME OF TREATMENT BERRY GROWTH T2: cane girdling T4: forchlorfenuron (Sitofex) T3: gibberellic acid (Berelex) Berry Ø: mm Berry Ø: mm

11 EXPERIMENTAL DESIGN Row 1 Row 2 Row 3 T3 T1 T2 T3 T1 T2 T3 T1 T2 T4 T2 T1 T4 T2 T1 T4 T2 T1 T1: control T2: cane girdling T3: gibberellic acid (Berelex) T4: forchlorfenuron (Sitofex) T1 T4 T3 T1 T4 T3 T1 T4 T3 60 berries/block (20 berries/vine) T2 T3 T4 T2 T3 T4 T2 T3 T4 180 berries/treatment

12 SAMPLING DATES July 1 August 5 September 3 October (harvest) July 30 July 20 August 4 September 2 October (harvest)

13 BERRIES ANALYSES morphological diameter (mm) length (mm) weight (g) chemical TSS ( Brix) ph TA (g/l tartaric acid) qualitative firmness (N) detachment force (N) colour metabolomic primary metabolites: sugars, organic acids, aminoacids

14 RESULTS 2011

15 Weight (g) Size (mm) Diameter Length b a 25.6 a 24.8 ab a ab Size parameters b 11.1 ab 10

16 TSS ( Brix) 18 ph TA (g/l tartaric acid) a 17.2 a 16.9 ab a b b 6.1 b 6.2 b Chemical parameters

17 Firmness (N) 4 BDF (N) Quality parameters 7 6 5

18 Color > Lightness L* C* h Control 41.9 ± 2.0 ab 11.6 ± ± 4.1 c Girdling 42.6 ± 2.3 a 11.2 ± ± 5.9 bc GA ± 1.9 ab 11.2 ± ± 3.9 a CPPU 41.3 ± 2.0 b 10.7 ± ± 4.4 ab

19 Yield/vine (kg) Cluster weight (g) a a a b a a a b 21 Zabadal e Budovc, 2006 Fidan et al.,

20 Metabolomics by NMR 2011 PCA applied to NMR data (generated from the regions of sugars and organic acids) Scores plot Loadings plot Black: control Blue: GA 3 Green: girdling Red: CPPU PC2 PC2 glucose Tartaric and malic acids PC1 fructose PC1 Girdling Higher FRUCTOSE content

21 Metabolomics by NMR 2011 PCA applied to NMR data (generated from the region of amino acids) Scores plot Loadings plot Black: control Blue: GA 3 Green: girdling Red: CPPU PC2 PC2 Arginine Proline Ethanol PC1 PC1

22 RESULTS 2012

23 Weight (g) Size (mm) Diameter Length b 27.5 a 27.2 a 27.2 a a 24.3 a 24.8 a b a 10.2 a a Size parameters b 8

24 TSS ( Brix) ph TA (g/l tartaric acid) a 3.28 a 3.20 b 3.28 a 3 Chemical parameters 2 1 0

25 Firmness (N) 4 BDF (N) Quality parameters 7 6 5

26 Color > Lightness Same color of the skin L* C* h Control 42.4 ± 2.0 B 10.6 ± ± 5.2 Girdling 43.1 ± 2.3 A 10.7 ± ± 6.4 GA ± 2.0 B 10.4 ± ± 5.1 CPPU 42.2 ± 2.0 B 10.6 ± ± 5.0

27 Yield/vine (kg) Cluster weight (g) b ab ab a a ab 22.9 ab b Zabadal e Budovc, 2006 Fidan et al.,

28 Metabolomics by NMR 2012 PCA applied to NMR data (generated from the regions of sugars and organic acids) Scores plot Loadings plot Black: control Blue: GA 3 Green: girdling Red: CPPU PC2 PC2 Glucose and fructose PC1 Tartaric and malic acids PC1 Girdling GA 3 CPPU Higher sugars/organic acids

29 Metabolomics by NMR 2012 PCA applied to NMR data (generated from the regions of sugars of amino and organic acids) acids) Scores plot Loadings plot Black: control Blue: GA 3 Green: girdling Red: CPPU PC2 PC2 PC1 PC1 No significant effects on amino acids composition

30 2011 vs 2012 Berry size larger in 2011 with respect to 2012 Control berries smaller than treated ones No differences for chemical parameters with the exception of ph, lower in 2012 (3.26) with respect to 2011 (3.74) Berry firmness and detachment forces similar in the two years Samples collected during 2011 possess higher sugars to acids ratio with respect to samples in 2012 Samples collected during 2011 possess higher arginine content with respect to samples in 2012

31 CONCLUSIONS Girdling and PGRs significantly affected size of the berry and yield/vine Limited influence on chemical parameters PGRs affected the color of the berry skin On the basis of our results and literature, an unappropriated use of PGRs may lead to negative qualitative characteristics Season is a key factor

32

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