Beijing, 4th ALLIUM Symposium, April 2004 G&H Influence of environmental and genetic factors on the...

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)


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)


bb


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


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.


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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


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


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


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


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)


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


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


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


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


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


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