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Beijing, 4th ALLIUM Symposium, April 2004 G&H
Influence of environmental and genetic factors on the alliin content of garlic bulbs
-
Odile HuchetteINRA, BV 86510, 21065 Dijon Cedex, France
European funded programme GARLIC AND HEALTH (FP5)
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
NH2 H
HOOC
COOH
SNH2
COOH
S-allyl-L-cysteine (SAC)
SNH2
COOH O
S
O
SH2O2
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)
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 temperature2. Influence of the light spectrum
Greenhouse (2002) : 3 cultivars and 3 sulphur fertilisation levels
Beijing, 4th ALLIUM Symposium, April 2004 G&H
nYear, Location Variety Planting date Harvest date Treatments (*)
Kg/ha
2000, France MC
PRI
MES
12-13 January
12-13 January
23 December
10 July
10 July
28 June
0-50-100-200
2000, Spain MC
PRI
MES
16-17 November
22-23 November
22-23 November
16 June
26 June
15 June
0-50-100-200
2001, France PRI 15 January 13 July 0-100-200-400
2001, Spain PRI 15 January August 0-100-200-400
(*) Sulphur fertilisation in the form of dry Calcium sulphate (gypsum) applied in two times (50% each)
Garlic Field experiments 2000-2001
Experimental design : 4 treatments and 4 replications in a randomised block design.
Beijing, 4th ALLIUM Symposium, April 2004 G&H
bb
Results obtained in France and Spain with MC, PRI and MESAlliin content measured in samples of garlic powder made with dehydrated bulbs (n=3).
Mean alliin content in Field Experiment 2000
MESMC
PRI
France
Spain
106.8 ns
90.2 ns
109.7 ns
38.9 c
81.2 a
66.3 b
0
20
40
60
80
100
120Alliin content in
bulbs (nmol/mg)
Variety
Statistical mean difference analysed by ANOVA was characterised and expressed as significant with a Newman-Keuls Test (p = 0.05).
influence of the genotype in France
Beijing, 4th ALLIUM Symposium, April 2004 G&H
bbFrance 2000 France 2001 Spain 2001Treatment
(S04 unit)
0 46.7 a 57.8 a 300.6 a
50 59.7 b - -
100 69.1 c 75.2 b 302.2 a
200 89.7 d 90.2 c 308.8 a
400 - 88.8 c 329.9 a
Mean 66.3 78.0 310.4
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
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)
Sulphur analysis on soil samples from experimental plots withoutany additional application of SO4 (2001) :
Sulphur in mg/ kg dry soil
25/08/00 14/05/01 20/06/01
France Spain France Spain FranceTotal 217 226 189 234 156
Water soluble 9 27.1 5,6 9,5 8
Hypothesis of 4 main factors according to the field trials
Soil :
Total and water soluble sulphur difference
Sulphur fertilisation : Not systematically
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
Beijing, 4th ALLIUM Symposium, April 2004 G&H
n
Influence of the genotype and sulphur fertilisation in greenhouse
0 S2 S
8 S
MES
MORPRI
ns
ns ns
nsns
ns
a ab bc
0
20
40
60
80
100
120Alliin content in bulbs (nmol/mg
FW)
Sulphur fertilisation level (in meq)
Variety
78.088.4
91.2
73.7
79.7
104.3
influence of the genotype
significant influence of the fertilisation level for MES
Alliin content of bulbs from varieties PRI, MOR and MES (2002)
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 1 month
22-24°C – F
Cold induction : 2 months3°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
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
0.00
0.05
0.10
0.15
0.20
0.25
22-24°C 27°CTemperature
Alliin content (nmol/mg FW)
S30
S60
Bulb formation under fluorescent light Sucrose : 30 versus 60 g/l Temperature : 22-24°C versus 27°C
positive influence of temperature not confirmed
Beijing, 4th ALLIUM Symposium, April 2004 G&H
nBulb 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
F+iF
MES
PRI
MOR
0.90 a
0.12 b
0.82 a
0.20 b0.18 b
0.17 b
0.0
0.2
0.4
0.6
0.8
1.0Alliin content in bulbs (nmol/mg
FW)
Light condition
Variety
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
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
Beijing, 4th ALLIUM Symposium, April 2004 G&H
Odile Huchette * , Christèle BellamyINRA, BV 86510, 21065 Dijon Cedex, France
Rémi KahaneCIRAD-FLHOR, TA50/PS4, bd de la Lironde, 34398 Montpellier Cedex 5, France
Jacques Auger, Ingrid ArnaultIRBI, Université François Rabelais, Avenue Monge, 37200 Tours, France
Influence of environmental and genetic factors on the alliin content of garlic bulbs
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