G&H October 2003, Münich WP3 / cultivation, P4 P4 : persons involved Odile Huchette, research engineer Christèle Bellamy, research technician Didier Perrin and Eric Vade, research assistants Influence of environmental conditions, genetic factors and mineral nutrition on the alliin content of garlic bulbs COOPD'OR R&D

G&H October 2003, Münich WP3 / cultivation, P4 G&H October 2003, Münich WP3 / cultivation, P4 Summary of the main conclusion of the experiments 2000-2002 Very high differences of alliin contents were observed in field trials between France and Spain : the soil as a factor of explanation but not the own In vitro, a major influence of i-light has been observed on different genotypes In controlled conditions, the alliin content has appeared to be linked to the genotype Garlic didn ’t seemed really affected by a carence of sulphur in natural conditions and in greenhouse An increasing effect of the sulphur fertilisation on the bulbs alliin content had to be confirmed COOPD'OR R&D

Influence of the intensity of the light G&H October 2003, Münich WP3 / cultivation, P4 To be confirmed in 2003 : Influence of the intensity of the light Effect of high dosis of sulphur (9 meq) Effect of nitrogen combined to sulphur Influence of S atmospheric COOPD'OR R&D

COOPD'OR R&D 2 experiments In vitro : G&H October 2003, Münich WP3 / cultivation, P4 Year 4 : what was done 2 experiments In vitro : Exp.III : Effect of various light conditions (intensity + quality) combined to 3 levels of sulphur in the medium CSO analysis achieved Exp.IV : Effect of S nutrition combined to N nutrition In vitro 2 experiments in greenhouse : 1. Effect of 3 nitrogen dosis (N, 2N and 3N) combined to a very high dosis of sulphur 2. Observation and analysis on garlic providing from cloves grown in different sulphur nutrition levels in 2002 CSO analysis in progress COOPD'OR R&D

- number of green leaves - lenght of the highest green leaf G&H October 2003, Münich WP3 / cultivation, P4 Experiments 2003  Observations - stage of the plant (in vitro : 1= growing / 2=beginning of bulbification / 3=maturation / 4=mature bulb) - number of green leaves - lenght of the highest green leaf  Measurements at harvest - weight, size and observation of the bulbs - alliin content (P5) GROWING BULB / ALLIIN COOPD'OR R&D

G&H October 2003, Münich WP3 / cultivation, P4 G&H October 2003, Münich WP3 / cultivation, P4 In vitro experiment III-2003 0 3 9 (in macro-elements) Gradient in SO4 meq 22-24°C - P = 16 hrs - F 1 month 22-24°C – P=16 hrs F = 9,50 µmol/s/m² 3 mediums (S- TS S++) 3 varieties : PRI + MES + MORl Cold induction 3°C - P = 10 hrs - F F+i = 9,5 µmol/s/m² F = 20 µmol/s/m² 2 months Effect of various light conditions (intensity + quality) and S nutrition

G&H October 2003, Münich WP3 / cultivation, P4 G&H October 2003, Münich WP3 / cultivation, P4 In vitro experiment III-2003 - Growth observed in all the S treatments but clear influence of absence of S in macro-nutrients on the development of the vitro-plants - Very low percentage of bulbs well-formed for Printanor, except under F+i light conditions and presence of sulphur

- Bulbs in treatment S- were quite big but soft G&H October 2003, Münich WP3 / cultivation, P4 In vitro experiment III-2003 - Bulbs in treatment S- were quite big but soft nd Alliin content Pri > Mor > Mes AlCSO couldn ’t be detected in bulbs under S- treatment (Pri) Effect of the S level (Pri) No additional effect of i-light in S++ conditions

Influence of the sulphur dosis confirmed for Printanor G&H October 2003, Münich WP3 / cultivation, P4 Conclusion of experiment III Positive effect of i-light confirmed for Morasol and Printanor in normal S conditions No positive influence of the intensity of light and of the temperature in these experimental conditions Influence of the sulphur dosis confirmed for Printanor Vitro-plants appeared more sensitive to an absence of S in macro-nutrients than plants grown in natural conditions or in greenhouse

22-24°C – P=16 hrs F+i – 9,50 µmol/s/m² G&H October 2003, Münich WP3 / cultivation, P4 In vitro experiment IV -2003 SO4 0 3 9 Gradient in SO4 and nitrogen meq (in macro-elements) 22-24°C - P = 16 hrs - F 1 month 9 9 medias varieties : PRI + MES + MORl + MORd Cold induction 3°C - P = 10 hrs - F 22-24°C – P=16 hrs F+i – 9,50 µmol/s/m² 2 months Nitrogen N 3N N 2N 3N N/2 N 2N 3N Effect of S nutrition combined to N nutrition

(I) Effect of N combined to an excess of S G&H October 2003, Münich WP3 / cultivation, P4 Greenhouse experiments 2003 (I) Effect of N combined to an excess of S 3 varieties : Printanor, Morasol, Messidrôme 3 mineral nutrition treatments : 3 levels of nitrogen combined to 9 meq of sulphur N, 2N, 3N 24 plants per treatment for each variety COOPD'OR R&D

G&H October 2003, Münich WP3 / cultivation, P4 Greenhouse experiment I

G&H October 2003, Münich WP3 / cultivation, P4 G&H October 2003, Münich WP3 / cultivation, P4 Greenhouse experiment I : 1st results on Messidrome => Less alliin measured when the dosis of N increases in presence of high dosis of S, but more GLUAlCS and IsoGLUAlCS COOPD'OR R&D

(II) Cloves from experiment 2002 G&H October 2003, Münich WP3 / cultivation, P4 Greenhouse experiment 2003 (II) Cloves from experiment 2002 COOPD'OR R&D

G&H October 2003, Münich WP3 / cultivation, P4 G&H October 2003, Münich WP3 / cultivation, P4 Greenhouse experiment II : growth of Morasol (36plants per treatment)

Influence of the S content of the bulb : G&H October 2003, Münich WP3 / cultivation, P4 Year 4 : still to be done Interaction SxN : CSO analysis on experiment IV in vitro CSO analysis on experiment I in greenhouse (Morasol, Printanor) Influence of the S content of the bulb : CSO analysis on experiment II in greenhouse (Printanor) => Should be available at the beginning of november COOPD'OR R&D

G&H October 2003, Münich WP3 / cultivation, P4 Milestones Year 4 ·        Experiments in vitro III and IV  Complementary experiment in greenhouse in year 4  Greenhouse cultivation of bulbs coming from the experiment 2002 COOPD'OR R&D

G&H October 2003, Münich WP3 / cultivation, P4 Deliverables P4 DP 2  Chinese garlic powder to be analysed DP 6  Garlic powder issued from various S-fertilisation in field condition DP 14  Chemical distinction between garlic accessions when cultivated in various sulphur nutrition regimes and under distinct temperatures in vitro DP 15  Physiological (growth traits) and chemical distinction (CSO) between accessions when cultivated in various sulphur nutrition regimes over two years DP 19  Chemical distinction (CSO) between accessions when cultivated in various sulphur (greenhouse) and mineral nutrition regimes (in vitro) DP 22  Paper on the interaction genotype x environment on the sulphur content of garlic plants grown in vitro , in greenhouse and in field conditions DP 30 Paper on the effect of combined sulphur, selenium and nitrogen nutrition on garlic growth, flavour precursor content and on biological potential of garlic for disease prevention DP 34  Chemical distinction (CSO) between garlic accessions when cultivated in various sulphur regimes (greenhouse) and mineral nutrition regimes (in vitro)