1. Beijing, 4th ALLIUM Symposium, April 2004 G&H Influence of environmental and genetic factors on the alliin content of garlic bulbs - Odile Huchette INRA, BV 86510, 21065 Dijon Cedex, France European funded programme GARLIC AND HEALTH (FP5)

2. Beijing, 4th ALLIUM Symposium, April 2004 G&H aa  -glutamyl-S-allyl-L-cysteine (GLUAlCS) S-allyl-L-cysteine sulfoxide  -glutamyl- transpeptidase Allicin Alliinase N O S NH 2 H HOOC COOH S NH 2 COOH S-allyl-L-cysteine (SAC) S NH 2 COOH O S O S H2O2H2O2 Alliin Factors affecting the quality of garlic ?  Numerous properties have been reported for garlic  Benefits probably linked to the high amount of organosulfur compounds, and particularly of thiosulfinates, found in garlic bulbs.  measured by HPLC ion-pairing method (Arnault et al, 2003) antifungal, antibacterial, antiviral, antithrombotic, antitumor, hypotensive, hypoglycemic, hypolipidemic (Augusti, 1996; Sato, 2000) therapeutic values related to cardiovascular diseases, cholesterol metabolism, atherosclerosis (Kik et al, 2001), and cancer (Le Bon and Siess, 2000)

3. Beijing, 4th ALLIUM Symposium, April 2004 G&H bb Influence of environmental and genetic factors on the alliin content of garlic bulbs  4 cultivars from 3 different varietal Groups in different experimental conditions : Morado (MC), Morasol (MOR), Printanor (PRI), Messidrôme (MES) Field trials : 2 locations (France, Spain) and 2 years (2000, 2001)  3 cultivars in 2000 (1 in 2001) and 4 sulphur fertilisation levels In vitro (Kahane et al, 1992a) : 3 cultivars 1. Influence of the temperature 2. Influence of the light spectrum Greenhouse (2002) : 3 cultivars and 3 sulphur fertilisation levels

4. Beijing, 4th ALLIUM Symposium, April 2004 G&H n Garlic Field experiments 2000-2001 Experimental design : 4 treatments and 4 replications in a randomised block design.

5. Beijing, 4th ALLIUM Symposium, April 2004 G&H bb Results obtained in France and Spain with MC, PRI and MES Alliin content measured in samples of garlic powder made with dehydrated bulbs (n=3). Mean alliin content in Field Experiment 2000  influence of the genotype in France

6. Beijing, 4th ALLIUM Symposium, April 2004 G&H bb Results from cultivar PRI Alliin content measured in samples of garlic powder made with dehydrated bulbs (n=3). Comparison of alliin contents from different fertilisation levels in both locations  influence of the fertilisation level in France  influence of the year and location Spain 2000 102.2

7. Beijing, 4th ALLIUM Symposium, April 2004 G&H bb Main factors affecting alliin content of garlic bulbs in the field ? Genotype : Higher alliin content in Spring planted varieties Temperature : 4°C warmer during the bulbing period in 2001 vs 2000 (Spain) Hypothesis of 4 main factors according to the field trials Soil : Total and water soluble sulphur difference Sulphur fertilisation : Not systematically

8. Beijing, 4th ALLIUM Symposium, April 2004 G&H n 3 varieties : PRI, MOR, MES 3 Sulphur fertilisation levels : 0 - 2 - 8 meq (macroelements) Influence of genotype and sulphur fertilisation in greenhouse 01/03 : Plantation in boxes with washed river sand 12 plants per box - 3 boxes per treatment 17/06 : Harvest of Messidrôme 10-11/07 : Harvest of Morasol 16-17/07 : Harvest of Printanor Mineral fertilisation and water applied by drop irrigation : 50 to 100 ml per day according to the stage of garlic

9. Beijing, 4th ALLIUM Symposium, April 2004 G&H n Influence of the genotype and sulphur fertilisation in greenhouse  influence of the genotype  significant influence of the fertilisation level for MES Alliin content of bulbs from varieties PRI, MOR and MES (2002)

10. Beijing, 4th ALLIUM Symposium, April 2004 G&H n Different genotypes (MOR, PRI, MES) exposed to various temperature and light spectrum conditions Experimental conditions in vitro 22-24°C - photoperiod 16 hrs - F 1month 22-24°C – F Cold induction : 2 months 3°C - photoperiod 10 hrs - F 27°C – F Growth medium (sucrose 30 to 60 g/l) photoperiod 16 hrs Garlic plantlets Bulbing period 22-24°C – F+i F : Fluorescent light F+i : Fluorescent + incandescent light

11. Beijing, 4th ALLIUM Symposium, April 2004 G&H Alliin content of bulbs from variety PRI grown in vitro on 2 different media under 2 culture temperature Influence of the temperature in vitro Bulb formation under fluorescent light Sucrose : 30 versus 60 g/l Temperature : 22-24°C versus 27°C  positive influence of temperature not confirmed

12. Beijing, 4th ALLIUM Symposium, April 2004 G&H n Bulb formation at 22-24°C with 60 g/l sucrose Light conditions : fluorescent (F) versus fluorescent + incandescent (F+i) Influence of the light conditions on 3 cultivars in vitro Alliin content of bulbs from varieties PRI, MOR and MES grown in vitro under 2 conditions of light spectrum  strong influence of the light spectrum for spring planted varieties

13. Beijing, 4th ALLIUM Symposium, April 2004 G&H n Conclusion To be confirmed : Influence of light intensity and temperature Role of mineral fertilisation in controlled conditions Slight increasing effect of the sulphur fertilisation, but no effect of the absence of sulphur fertilisation Strong influence of the genotype observed in the field and in controlled conditions Major influence of the light spectrum stressed on Spring planted varieties grown in vitro, but effect of temperature not confirmed Very high differences of alliin contents observed in the field trials between France and Spain  strong influence of the environment, only partially explained by the sulphur status of the soil

14. Beijing, 4th ALLIUM Symposium, April 2004 G&H Odile Huchette *, Christèle Bellamy INRA, BV 86510, 21065 Dijon Cedex, France Rémi Kahane CIRAD-FLHOR, TA50/PS4, bd de la Lironde, 34398 Montpellier Cedex 5, France Jacques Auger, Ingrid Arnault IRBI, Université François Rabelais, Avenue Monge, 37200 Tours, France Influence of environmental and genetic factors on the alliin content of garlic bulbs THANK YOU !