1. COOPD’OR, IRBI S.T.L., UNISEM, Univ. Cordoba
G&H WP3 : Cultivation
COOPD’OR, IRBI
S.T.L., UNISEM, Univ. Cordoba
Final meeting Leiden 01/04

2. Garlic cultivation : Plans and objectives
What did we expect from this workpackage ?
a standard composition in Alliin per clove
distinction between varieties
distinction between practices
S-fertilisation
Environmental effects (light, temperature)
N and S
Se and S
powder material for Health experimentation
indications for tuning garlic quality in the field
Final meeting Leiden 01/04

3. Garlic cultivation : Plans and objectives
What really happened during the project ?
the analytical method has been modified
not only Alliin had to be analysed
the field trials were hardly comparable between locations and dates
powder was not all that easy to handle
plants in vitro should have been multiplied and induced to bulb before analysis
experiments in greenhouse were fine but Se could not be tested
Final meeting Leiden 01/04

4. Garlic cultivation : Plans and schedule
Year 1
Initiation of in vitro work
Analytical method for Alliin in whole bulbs or in powder
Field trial in two locations (France and Spain)
CSO analysis of the field trial products before and after processing into powder
Transfer to the Health partners
Year 2
In vitro : temperature effect on CSO
In vitro : S nutrition effect on CSO (normal and S-enriched)
Preparation of greenhouse experiments
Field trial in two locations (France and Spain)
CSO analysis of the field trial products before and after processing into powder
Transfer to the Health partners
Final meeting Leiden 01/04

5. Garlic cultivation : Plans and schedule
Year 3
In vitro : light and S N nutrition effects on CSO
Greenhouse experiments with 4 genotypes and high S nutrition for producing S-enriched garlic bulbs ; CSO analysis
CSO comparison between virus-free and virus-contaminated plants
Year 4
In vitro : S and N nutrition effects on CSO (not Se)
In vitro : plants from embryogenesis also tested for light and S nutrition effects on CSO
Greenhouse cultivation of garlic bulbs from year 3
Greenhouse experiments with one genotype and various S and N nutrition regimes
Chemical analysis before and after processing into powder
Transfer to the Health partners
Final meeting Leiden 01/04

6. Main results per experimental tool
Analytical methods
In vitro
Greenhouse
Field
Final meeting Leiden 01/04

7. Main results in analytical methods
Pico Tag method quantifies only the summary of Alliin and GLUACS
A new HPLC method (Lichtwer's method) was validated
Final meeting Leiden 01/04

8. Main results in analytical methods
0,00
0,02
0,04
0,06
0,08
0,10
10,00
20,00
30,00
40,00
50,00
60,00
(+)-Alliin +GLUACS
Pico Tag method
Lichtwer's method
Alliin and two dipeptides have to
be measured to give an good
image of garlic potential flavour
0,00
0,20
0,40
5,00
10,00
15,00
20,00
25,00
GLUACS
(-)-alliin
(+)-alliin
Final meeting Leiden 01/04

9. Main results in vitro Functional tool
Effect of light quality has been stressed, to be further analyzed
S nutrition slightly effective
Strong Genotype x Environment interactions
Limits of this tool
Final meeting Leiden 01/04

10. Main results in vitro Functional tool
Possible independent testing of environmental factors (temperature, daylength, light spectrum quality, light intensity) or trophic factors (carbohydrates, S source, N source) on homogeneous plant material.
Final meeting Leiden 01/04

11. Main results in vitro Effect of light quality
Additional far red light (incandescence) induced higher amounts of CSO stored in the garlic bulbs (here Printanor).
3,5
0.8
Sucrose (g/l)
3,0
0.7 30 60
120
0.6
2,5
0.5
2,0
Mean bulb weight (g)
Alliin (mmol/mg FW)
0.4
1,5
0.3
1,0
0.2
0,5
0.1
0,0
0.0 F
F+i F
F+i
Final meeting Leiden 01/04

12. Main results in vitro Effect of light quality
Additional far red light (incandescence) may induce higher amounts of CSO stored in the garlic bulbs of certain varieties.
Needs to be further studied.
MESSIDROME
PRINTANOR 1 2 3 4 5 6 7 8
S30
S60
S30_3
S60_3
S120_3
Alliin (mmol/mg FW) F
F+i
S=sucrose (g/l)
Final meeting Leiden 01/04

13. Main results in vitro
S nutrition has not effect on plant growth, but on alliin concentration
Alliin concentration
(nmol/mg FW)
MORASOL
PRINTANOR
MESSIDROME
MORADO
Mean bulb weight (g)
0,0
0,1
0,2
0,3
0,4
0,5
0,6
2S_F+i
1S_F+i
0,8
1,0
Final meeting Leiden 01/04

14. Main results in vitro S nutrition affects N valorisation for alliin4 5 6 7
S++ (3S)
N/2 3N N 2N 1 2 3
S normal 8 9 S-
Alliin concentration
(nmol/mg FW)
0,0
1,0
2,0
3,0
4,0
5,0
6,0
7,0
PRINTANOR MORASOL MORADO MESSIDROME
Final meeting Leiden 01/04

15. Main results in vitro Genotype x Environment interactions
0,5 1
1,5 2
2,5 3
3,5 4
S- F
TS F
S+ F
S- 2F
TS 2F
S+ 2F
S- F+i
TS F+i
S+ F+i
PRINTANOR MORASOL MESSIDROME
Alliin concentration
(nmol/mg FW)
Final meeting Leiden 01/04

16. Main results in vitro Genotype x Environment interactions
0,5 1
1,5 2
2,5 3
3,5 4
4,5 5
S- F
TS F
S++ F
S- 2F
TS 2F
S++ 2F
S- F+i
TS F+i
S++ F+i 2 4 6 8 10 12 14 16 18 20
S- F
TS F
S++ F
S- 2F
TS 2F
S++ 2F
S- F+i
TS F+i
S++ F+i
(Iso)-GluAlCSO
(nmol/mg FW)
PRINTANOR MORASOL MESSIDROME
Final meeting Leiden 01/04

17. Main results in vitro Limits of this tool small amount of material
heterogeneity in the bulbing response
representativity to real bulbing ?
representativity to real flavour ?
Final meeting Leiden 01/04

18. Main results in greenhouse
Genotype distinction
S nutrition efficiency on CSOs
Little S/N interactions
Limits of this tool
Final meeting Leiden 01/04

19. Main results in greenhouse
Genotype distinction 2 4 6 8 10 12 14 16
5/3
25/3
14/4
4/5
24/5
13/6
3/7
Dates
Number of leaves
Printanor
Morasol
Messidrôme
MES
MOR
PRI 59 75 91
107
123
Alliin (nmol/mg FW)
Final meeting Leiden 01/04

20. Main results in greenhouse
S nutrition not effective on CSOs
2002
AlCSO
75.1
83.4
80.7
79.7
92.6
107.8
111.9
104.1
66.2
73.9
80.9
73.7
GLU
36.7
30.8
33.1
33.5
10.9
10.2
11.7
32.9
44.6
45.4
41.0
IsoGLU
33.8
40.3
36.5
36.9
26.1
35.6
34.8
32.2
34.5
49.7
45.7
43.3
Total
150.1
147.2
158.0
2003
AlCSO
GLU
IsoGLU
Total
61.9
20.6
34.3
60.7
20.1
34.1
59.3
19.3
34.2
60.6
20.0
114.8
73.4
9.2
29.5
68.6
11.1
33.0
92.8
12.9
29.9
78.3
11.0
30.8
120.1
53.4
20.7
42.1
52.4
18.8
32.5
30.2
51.1
53.1
23.2
41.9
118.2
Origin
MOR_S-
MOR_TS
MOR_S++
Mean MOR
PRI_S-
PRI_TS
PRI_S++
Mean PRI
MES_S-
MES_TS
MES_S++
Mean MES
Final meeting Leiden 01/04

21. Main results in greenhouse
Little S/N interactions
MESSIDROME
S++N
S++N+
S++N++ 20 30 40 50 60 70 80 90
AlCSO
GLUAlCS
IsoGLUAlCS
Concentrations
(nmol/mg FW)
Final meeting Leiden 01/04

22. Main results in greenhouse
Limits of this tool
- small number of treatments
- homogeneity ?
- representativity to real flavour ?
Final meeting Leiden 01/04

23. Main results in field Genotype distinction
Which environmental effect ?
Powders processing and handling
Limits of field trials
Final meeting Leiden 01/04

24. Main results in field
No effect of S fertilization on bulb yield (g/bulb, Printanor)
Spain
2000
2001
98.0
59.5
102.2 -
101.3
58.1
103.2
52.4
59.6 nd
101.2
57.4
Treatment
(S0 4
unit) 50
100
200
400
2000
Mean
France
2000
2001
65.8
89.9
63.2 -
60.0
86.1
62.5
84.7
87.3
62.9
87.0
nd : not determined
Final meeting Leiden 01/04

25. Main results in field
Small effect of S fertilization on alliin (nmol/mg FW, Printanor)
Treatment
(S0 4
unit) 50
100
200
400
2000
Mean
France
2000
30,8
37,5
40,1
47,1 -
38,9
Spain
2000 nd
2001
300,6
302,2
308,8
329,9
325,8
313,5
109,7
2001
57,8
75,2
90,2
88,8 -
78,0
nd : not determined
Final meeting Leiden 01/04

26. Main results in field Which environmental effect ? - Soil analysis
France
Spain
Treatment
(S04 unit) 25/08/00 21/02/01 14/05/01 20/06/01 14/08/01
0 Total S
Water soluble ,
100 Total S
Water soluble ,
200 Total S
Water soluble ,
400 Total S
Water soluble ,
14/05/01
234
9,5
249
10,1
238
8,7
244
8,4
Final meeting Leiden 01/04

27. Main results in field Which environmental effect ?
Rainfall 40
300
year 1
2001*
Rainfall 35
2000*
250
year 2 30
T° av
year 1
200 25
Temperatures (°C)
Rainfall (mm)
T° av
year 2 20
150 15
100 10 50
- Climatic conditions
in Spain 5
March
April
Mai
June
July
November
December
January
February
Final meeting Leiden 01/04

28. Main results in field Powders processing and handling
kg of powders produced, stored, analysed and supplied to Health participants
strong differences in CSO composition between powder lots, allowing biological distinct effects
constant evolution of CSO composition along time
Final meeting Leiden 01/04

29. Main results in field Limits of field trials
multiple and uncontrolled environmental factors (soil, climate, human intervention…)
bulb material handling, processing and storing
on-farm versus on-station experiments
Final meeting Leiden 01/04

30. Deliverables DP2. Chinese garlic powder analysis (as control) (P5)
DP6. Garlic powder from bulbs (field trial 2000) with distinct varieties and S treatments (P4, P5)
Garlic powder from bulbs (field trial 2001) of one variety with distinct S treatments (P4, P5)
DP14. Physiological and chemical distinctions (CSO) between garlic varieties when cultivated in various S-nutrition regimes and under distinct environments in vitro (P4, P5)
DP15. Physiological and chemical distinction (CSO) in garlic when cultivated in various S-nutrition regimes (field trials) (P4, P5)
Final meeting Leiden 01/04

31. Deliverables
DP19. Garlic powder from bulbs (greenhouse grown) with distinct CSO contents for biological experiments (P4)
DP22. Paper on the interaction genotype x environment on CSO and total S contents of garlic plants grown in vitro, in greenhouse and in field conditions (P4, P5, P6 + P2)
DP34. Chemical distinction (CSO) between garlic varieties when cultivated in various S regimes (greenhouse) and mineral nutrition regimes (in vitro) (P4, P5)
DP30. Paper on the effect of combined sulphur and nitrogen (not selenium) nutrition on garlic growth, flavour precursor content (and not on biological potential of garlic for disease prevention)
(not yet achieved)
Final meeting Leiden 01/04

32. Positive and Negative points
Key lessons
Positive and
Negative points
Final meeting Leiden 01/04

33. Positive Points Email reports/general communication Shared experiments
Shared materials and protocols between Plant and Health partners
Final meeting Leiden 01/04

34. Negative Points Little interaction with private partners
low input from STL
low output for UNISEM
Punctual miscommunication
labelling samples from the field
unusable data due to different methods in CSO analysis
Smooth advertising of the project and results
Final meeting Leiden 01/04

35. Perspectives
Publications, diffusion of the results (symposium, partnership, …)
Applied impact for partners (UNISEM)
New contracts (valorising our experience) with private partners
Final meeting Leiden 01/04

36. Questions or comments …
… are welcome !
Final meeting Leiden 01/04