COOPD’OR, IRBI S.T.L., UNISEM, Univ. Cordoba G&H WP3 : Cultivation COOPD’OR, IRBI S.T.L., UNISEM, Univ. Cordoba Final meeting Leiden 01/04

Garlic cultivation : Plans and objectives What did we expect from this workpackage ? a standard composition in Alliin per clove distinction between varieties distinction between practices S-fertilisation Environmental effects (light, temperature) N and S Se and S powder material for Health experimentation indications for tuning garlic quality in the field Final meeting Leiden 01/04

Garlic cultivation : Plans and objectives What really happened during the project ? the analytical method has been modified not only Alliin had to be analysed the field trials were hardly comparable between locations and dates powder was not all that easy to handle plants in vitro should have been multiplied and induced to bulb before analysis experiments in greenhouse were fine but Se could not be tested Final meeting Leiden 01/04

Garlic cultivation : Plans and schedule Year 1 Initiation of in vitro work Analytical method for Alliin in whole bulbs or in powder Field trial in two locations (France and Spain) CSO analysis of the field trial products before and after processing into powder Transfer to the Health partners Year 2 In vitro : temperature effect on CSO In vitro : S nutrition effect on CSO (normal and S-enriched) Preparation of greenhouse experiments Field trial in two locations (France and Spain) CSO analysis of the field trial products before and after processing into powder Transfer to the Health partners Final meeting Leiden 01/04

Garlic cultivation : Plans and schedule Year 3 In vitro : light and S N nutrition effects on CSO Greenhouse experiments with 4 genotypes and high S nutrition for producing S-enriched garlic bulbs ; CSO analysis CSO comparison between virus-free and virus-contaminated plants Year 4 In vitro : S and N nutrition effects on CSO (not Se) In vitro : plants from embryogenesis also tested for light and S nutrition effects on CSO Greenhouse cultivation of garlic bulbs from year 3 Greenhouse experiments with one genotype and various S and N nutrition regimes Chemical analysis before and after processing into powder Transfer to the Health partners Final meeting Leiden 01/04

Main results per experimental tool Analytical methods In vitro Greenhouse Field Final meeting Leiden 01/04

Main results in analytical methods Pico Tag method quantifies only the summary of Alliin and GLUACS A new HPLC method (Lichtwer's method) was validated Final meeting Leiden 01/04

Main results in analytical methods 0,00 0,02 0,04 0,06 0,08 0,10 10,00 20,00 30,00 40,00 50,00 60,00 (+)-Alliin +GLUACS Pico Tag method Lichtwer's method Alliin and two dipeptides have to be measured to give an good image of garlic potential flavour 0,00 0,20 0,40 5,00 10,00 15,00 20,00 25,00 GLUACS (-)-alliin (+)-alliin Final meeting Leiden 01/04

Main results in vitro Functional tool Effect of light quality has been stressed, to be further analyzed S nutrition slightly effective Strong Genotype x Environment interactions Limits of this tool Final meeting Leiden 01/04

Main results in vitro Functional tool Possible independent testing of environmental factors (temperature, daylength, light spectrum quality, light intensity) or trophic factors (carbohydrates, S source, N source) on homogeneous plant material. Final meeting Leiden 01/04

Main results in vitro Effect of light quality Additional far red light (incandescence) induced higher amounts of CSO stored in the garlic bulbs (here Printanor). 3,5 0.8 Sucrose (g/l) 3,0 0.7 30 60 120 0.6 2,5 0.5 2,0 Mean bulb weight (g) Alliin (mmol/mg FW) 0.4 1,5 0.3 1,0 0.2 0,5 0.1 0,0 0.0 F F+i F F+i Final meeting Leiden 01/04

Main results in vitro Effect of light quality Additional far red light (incandescence) may induce higher amounts of CSO stored in the garlic bulbs of certain varieties. Needs to be further studied. MESSIDROME PRINTANOR 1 2 3 4 5 6 7 8 S30 S60 S30_3 S60_3 S120_3 Alliin (mmol/mg FW) F F+i S=sucrose (g/l) Final meeting Leiden 01/04

Main results in vitro S nutrition has not effect on plant growth, but on alliin concentration Alliin concentration (nmol/mg FW) MORASOL PRINTANOR MESSIDROME MORADO Mean bulb weight (g) 0,0 0,1 0,2 0,3 0,4 0,5 0,6 2S_F+i 1S_F+i 0,8 1,0 Final meeting Leiden 01/04

Main results in vitro S nutrition affects N valorisation for alliin 4 5 6 7 S++ (3S) N/2 3N N 2N 1 2 3 S normal 8 9 S- Alliin concentration (nmol/mg FW) 0,0 1,0 2,0 3,0 4,0 5,0 6,0 7,0 PRINTANOR MORASOL MORADO MESSIDROME Final meeting Leiden 01/04

Main results in vitro Genotype x Environment interactions 0,5 1 1,5 2 2,5 3 3,5 4 S- F TS F S+ F S- 2F TS 2F S+ 2F S- F+i TS F+i S+ F+i PRINTANOR MORASOL MESSIDROME Alliin concentration (nmol/mg FW) Final meeting Leiden 01/04

Main results in vitro Genotype x Environment interactions 0,5 1 1,5 2 2,5 3 3,5 4 4,5 5 S- F TS F S++ F S- 2F TS 2F S++ 2F S- F+i TS F+i S++ F+i 2 4 6 8 10 12 14 16 18 20 S- F TS F S++ F S- 2F TS 2F S++ 2F S- F+i TS F+i S++ F+i (Iso)-GluAlCSO (nmol/mg FW) PRINTANOR MORASOL MESSIDROME Final meeting Leiden 01/04

Main results in vitro Limits of this tool small amount of material heterogeneity in the bulbing response representativity to real bulbing ? representativity to real flavour ? Final meeting Leiden 01/04

Main results in greenhouse Genotype distinction S nutrition efficiency on CSOs Little S/N interactions Limits of this tool Final meeting Leiden 01/04

Main results in greenhouse Genotype distinction 2 4 6 8 10 12 14 16 5/3 25/3 14/4 4/5 24/5 13/6 3/7 Dates Number of leaves Printanor Morasol Messidrôme MES MOR PRI 59 75 91 107 123 Alliin (nmol/mg FW) Final meeting Leiden 01/04

Main results in greenhouse S nutrition not effective on CSOs 2002 AlCSO 75.1 83.4 80.7 79.7 92.6 107.8 111.9 104.1 66.2 73.9 80.9 73.7 GLU 36.7 30.8 33.1 33.5 10.9 10.2 11.7 32.9 44.6 45.4 41.0 IsoGLU 33.8 40.3 36.5 36.9 26.1 35.6 34.8 32.2 34.5 49.7 45.7 43.3 Total   150.1 147.2 158.0 2003 AlCSO GLU IsoGLU Total 61.9 20.6 34.3   60.7 20.1 34.1 59.3 19.3 34.2 60.6 20.0 114.8 73.4 9.2 29.5 68.6 11.1 33.0 92.8 12.9 29.9 78.3 11.0 30.8 120.1 53.4 20.7 42.1 52.4 18.8 32.5 30.2 51.1 53.1 23.2 41.9 118.2 Origin MOR_S- MOR_TS MOR_S++ Mean MOR PRI_S- PRI_TS PRI_S++ Mean PRI MES_S- MES_TS MES_S++ Mean MES Final meeting Leiden 01/04

Main results in greenhouse Little S/N interactions MESSIDROME S++N S++N+ S++N++ 20 30 40 50 60 70 80 90 AlCSO GLUAlCS IsoGLUAlCS Concentrations (nmol/mg FW) Final meeting Leiden 01/04

Main results in greenhouse Limits of this tool - small number of treatments - homogeneity ? - representativity to real flavour ? Final meeting Leiden 01/04

Main results in field Genotype distinction Which environmental effect ? Powders processing and handling Limits of field trials Final meeting Leiden 01/04

Main results in field No effect of S fertilization on bulb yield (g/bulb, Printanor) Spain 2000 2001 98.0 59.5 102.2 - 101.3 58.1 103.2 52.4 59.6 nd 101.2 57.4 Treatment (S0 4 unit) 50 100 200 400 2000 Mean France 2000 2001 65.8 89.9 63.2 - 60.0 86.1 62.5 84.7 87.3 62.9 87.0 nd : not determined Final meeting Leiden 01/04

Main results in field Small effect of S fertilization on alliin (nmol/mg FW, Printanor) Treatment (S0 4 unit) 50 100 200 400 2000 Mean France 2000 30,8 37,5 40,1 47,1 - 38,9 Spain 2000 nd 2001 300,6 302,2 308,8 329,9 325,8 313,5 109,7 2001 57,8 75,2 90,2 88,8 - 78,0 nd : not determined Final meeting Leiden 01/04

Main results in field Which environmental effect ? - Soil analysis France Spain Treatment (S04 unit) 25/08/00 21/02/01 14/05/01 20/06/01 14/08/01 0 Total S 217 149 189 156 208 Water soluble 9 7 5,6 8 10 100 Total S 172 187 186 Water soluble 9,7 11 9 200 Total S 168 127 213 Water soluble 12,2 16 15 400 Total S 188 114 185 Water soluble 20,8 22 20 14/05/01 234 9,5 249 10,1 238 8,7 244 8,4 Final meeting Leiden 01/04

Main results in field Which environmental effect ? Rainfall 40 300 year 1 2001* Rainfall 35 2000* 250 year 2 30 T° av year 1 200 25 Temperatures (°C) Rainfall (mm) T° av year 2 20 150 15 100 10 50 - Climatic conditions in Spain 5 March April Mai June July November December January February Final meeting Leiden 01/04

Main results in field Powders processing and handling kg of powders produced, stored, analysed and supplied to Health participants strong differences in CSO composition between powder lots, allowing biological distinct effects constant evolution of CSO composition along time Final meeting Leiden 01/04

Main results in field Limits of field trials multiple and uncontrolled environmental factors (soil, climate, human intervention…) bulb material handling, processing and storing on-farm versus on-station experiments Final meeting Leiden 01/04

Deliverables DP2. Chinese garlic powder analysis (as control) (P5) DP6. Garlic powder from bulbs (field trial 2000) with distinct varieties and S treatments (P4, P5) Garlic powder from bulbs (field trial 2001) of one variety with distinct S treatments (P4, P5) DP14. Physiological and chemical distinctions (CSO) between garlic varieties when cultivated in various S-nutrition regimes and under distinct environments in vitro (P4, P5) DP15. Physiological and chemical distinction (CSO) in garlic when cultivated in various S-nutrition regimes (field trials) (P4, P5) Final meeting Leiden 01/04

Deliverables DP19. Garlic powder from bulbs (greenhouse grown) with distinct CSO contents for biological experiments (P4) DP22. Paper on the interaction genotype x environment on CSO and total S contents of garlic plants grown in vitro, in greenhouse and in field conditions (P4, P5, P6 + P2) DP34. Chemical distinction (CSO) between garlic varieties when cultivated in various S regimes (greenhouse) and mineral nutrition regimes (in vitro) (P4, P5) DP30. Paper on the effect of combined sulphur and nitrogen (not selenium) nutrition on garlic growth, flavour precursor content (and not on biological potential of garlic for disease prevention) (not yet achieved) Final meeting Leiden 01/04

Positive and Negative points Key lessons Positive and Negative points Final meeting Leiden 01/04

Positive Points Email reports/general communication Shared experiments Shared materials and protocols between Plant and Health partners Final meeting Leiden 01/04

Negative Points Little interaction with private partners low input from STL low output for UNISEM Punctual miscommunication labelling samples from the field unusable data due to different methods in CSO analysis Smooth advertising of the project and results Final meeting Leiden 01/04

Perspectives Publications, diffusion of the results (symposium, partnership, …) Applied impact for partners (UNISEM) New contracts (valorising our experience) with private partners Final meeting Leiden 01/04

Questions or comments … … are welcome ! Final meeting Leiden 01/04