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G&H WP3 : Cultivation

G&H WP3 : Cultivation. COOPD’OR, IRBI S.T.L., UNISEM, Univ. Cordoba. 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

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G&H WP3 : Cultivation

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

  2. 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

  3. 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

  4. 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

  5. Garlic cultivation : Plans and schedule • Year 3 • In vitro : light and S N nutrition effects on CSO • Greenhouse experiments with 4 genotypesand 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 genotypeand various S and N nutrition regimes • Chemical analysis before and after processing into powder • Transfer to the Health partners Final meeting Leiden 01/04

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

  7. 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

  8. 0,10 (+)-Alliin +GLUACS 0,08 0,06 0,04 0,02 0,00 0,00 10,00 20,00 30,00 40,00 50,00 60,00 GLUACS 0,40 (-)-alliin (+)-alliin 0,20 0,00 0,00 5,00 10,00 15,00 20,00 25,00 Main results in analytical methods • Pico Tag method • Lichtwer's method Alliin and two dipeptides have to be measured to give an good image of garlic potential flavour Final meeting Leiden 01/04

  9. 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

  10. 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

  11. 30 60 120 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) 0.7 3,0 0.6 2,5 0.5 Mean bulb weight (g) 2,0 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

  12. F F+i 8 MESSIDROME 7 PRINTANOR 6 5 Alliin (mmol/mg FW) 4 3 2 1 0 S30 S60 S30_3 S60_3 S120_3 Needs to be further studied. S=sucrose (g/l) 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. Final meeting Leiden 01/04

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

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

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

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

  17. 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

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

  19. 123 107 16 14 Alliin (nmol/mg FW) 91 12 Printanor 10 Morasol Number of leaves 8 75 6 Messidrôme 4 2 59 MES MOR PRI 0 24/5 13/6 3/7 5/3 25/3 14/4 4/5 Dates Main results in greenhouse • Genotype distinction Final meeting Leiden 01/04

  20. 2002 Origin 2003 AlCSO MOR_S- AlCSO GLU GLU IsoGLU IsoGLU Total Total 75.1 61.9 MOR_TS 36.7 20.6 34.3 33.8 60.7 20.1 34.1 MOR_S++ 83.4 30.8 40.3 59.3 19.3 34.2 Mean MOR 80.7 33.1 36.5 60.6 20.0 34.2 114.8 PRI_S- 79.7 33.5 36.9 150.1 73.4 9.2 29.5 PRI_TS 92.6 10.9 26.1 68.6 11.1 33.0 PRI_S++ 107.8 10.2 35.6 92.8 12.9 29.9 Mean PRI 111.9 11.7 34.8 78.3 11.0 30.8 120.1 MES_S- 104.1 10.9 32.2 147.2 53.4 20.7 42.1 MES_TS 66.2 32.9 34.5 52.4 18.8 32.5 MES_S++ 53.4 30.2 51.1 73.9 44.6 49.7 Mean MES 53.1 23.2 41.9 118.2 80.9 45.4 45.7 73.7 41.0 43.3 158.0 Main results in greenhouse • S nutrition not effective on CSOs Final meeting Leiden 01/04

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

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

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

  24. Treatment Spain France (S0 unit) 2000 2001 2000 2001 4 0 65.8 89.9 98.0 59.5 50 63.2 - 102.2 - 100 60.0 86.1 101.3 58.1 200 62.5 84.7 103.2 52.4 400 - 87.3 - 59.6 2000 - - - nd Mean 62.9 87.0 101.2 57.4 Main results in field • No effect of S fertilization on bulb yield (g/bulb, Printanor) nd : not determined Final meeting Leiden 01/04

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

  26. 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 100Total 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

  27. 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 Rainfall (mm) Temperatures (°C) T° av year 2 20 150 15 100 10 -Climatic conditions in Spain 50 5 0 0 July June Mai April March January February December November Final meeting Leiden 01/04

  28. 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

  29. 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

  30. 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

  31. 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

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

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

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

  35. 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

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

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