Influence of environmental and genetic factors on the alliin content of garlic bulbs -

1. European funded programme GARLIC AND HEALTH (FP5) Influence of environmental and genetic factors on the alliin content of garlic bulbs - Odile Huchette INRA, BV 86510, 21065 Dijon Cedex, France

2. Alliin H N H O 2 H2O2 H N N Alliinase H N S 2 2 -glutamyl-transpeptidase S H O O C S S S C O O H O C O O H C O O H O S-allyl-L-cysteine (SAC) S-allyl-L-cysteine sulfoxide Allicin -glutamyl-S-allyl-L-cysteine (GLUAlCS) Factors affecting the quality of garlic ?  Numerous properties have been reported for garlic • 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) aa  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)

3. 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) bb Field trials : 2 locations (France, Spain) and 2 years (2000, 2001)  3 cultivars in 2000 (1 in 2001) and 4 sulphur fertilisation levels Greenhouse (2002) : 3 cultivars and 3 sulphur fertilisation levels In vitro(Kahane et al, 1992a) : 3 cultivars 1. Influence of the temperature 2. Influence of the light spectrum

4. Garlic Field experiments 2000-2001 Experimental design : 4 treatments and 4 replications in a randomised block design. n

5. Mean alliin content in Field Experiment 2000 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). bb  influence of the genotype in France

6. Spain 2000 102.2 Comparison of alliin contents from different fertilisation levels in both locations Results from cultivar PRI Alliin content measured in samples of garlic powder made with dehydrated bulbs (n=3). bb  influence of the fertilisation level in France  influence of the year and location

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

8. 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 Influence of genotype and sulphur fertilisation in greenhouse 3 varieties : PRI, MOR, MES 3 Sulphur fertilisation levels : 0 - 2 - 8 meq (macroelements) n

9. Influence of the genotype and sulphur fertilisation in greenhouse Alliin content of bulbs from varieties PRI, MOR and MES (2002) n  influence of the genotype  significant influence of the fertilisation level for MES

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

11. Influence of the temperature in vitro Alliin content of bulbs from variety PRI grown in vitro on 2 different media under 2 culture temperature 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. 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 n Bulb formation at 22-24°C with 60 g/l sucrose Light conditions : fluorescent (F) versus fluorescent + incandescent (F+i)  strong influence of the light spectrum for spring planted varieties

13. 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 Conclusion • Strong influence of the genotype observed in the field and in controlled conditions n • Major influence of the light spectrum stressed on Spring planted varieties grown in vitro, but effect of temperature not confirmed • Slight increasing effect of the sulphur fertilisation, but no effect of the absence of sulphur fertilisation • To be confirmed : • Influence of light intensity and temperature • Role of mineral fertilisation in controlled conditions

14. Influence of environmental and genetic factors on the alliin content of garlic bulbs THANK YOU ! 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