Irish Phytochemical Food Network
Systems Comparison Trial
Reilly, K., Finn. L., Valverde, J., Grant, J., Gaffney, M., Brunton, N.
OGI Farm Walk 10th July 2012
email: kim.reilly@teagasc.ie
Irish Phytochemical Food Network
A FIRM funded project set up in 2008 to study
phytochemicals in Irish grown fruit and vegetables
“from farm to fork”. It is made up of researchers
from Teagasc Kinsealy and AFRC, UCC, UCD,
UCL, DIT and UCC.
phytochemicals = “non-nutrient compounds
produced by plants that have biological activity
against chronic disease”. AKA “plant bioactives”
Funding body:
Members:
Teagasc Kinsealy : Work-package 3
•
•
•
•
•
Variety
Plant nutrition
Plant maturity
Tissue type
Plant spacing
• Conventional vs. Organic
Production Practices
Broccoli Field Trial 2008
Onion – quercetin
Broccoli- glucosinolates
Carrot - polyacetylenes
Types of study:
1) Basket survey e.g. Meyer et al. 2008
2) Paired Farm Survey e.g. Lester et al. 2007
3) Field trial e.g. Soltoft et al. 2010
Suggested criteria (Dangour et al. 2009)
1) specification of the cultivar of crop or breed of livestock
2) a statement of which nutrient or other nutritionally relevant
substance was analyzed
3) a description of the laboratory analytic methods used
4) a statement of the methods used for statistical analyses
5) a clear definition of the organic production methods, including
the name of the organic certification body.
Systems Comparison Trial:
Why is the systems trial important?
Public Good:
• Producing healthy food
• Producing sustainable food
• Encourage awareness
of F&V as healthy food
Grower and Industry:
• What inputs are important?
• Encourage consumption of F&V
• Marketing
Academic:
• Understand factors that affect
phytochemicals in vegetables
• Provide evidence for farming
inputs or practices
• Generate peer reviewed
publications
Systems Comparison trial
Objective:
To investigate the effect of, and interaction between, production
system components - a) soil management and b) pest control
measures, that differ between organic and conventional systems, on
phytochemical accumulation in 2 varieties each of carrot, broccoli
and onion.
2x2x2 factorial split plot design with 4 replicates
Main-plot: 2 varieties of each crop (V1, V2)
Sub-plot: 2 Levels of soil treatment a) Organic Soil treatment (OS)
b) Conventional Soil treatment (CS)
Sub sub plot: 2 levels of pest control a) Organic pest control (OP)
b) Conventional pest control (CP)
This gives 8 experimental treatments for each crop
1) V1 OS OP
2) V1 OS CP
3) V1 CS OP
4) V1 CS CP
5) V2 OS OP
6) V2 OS CP
7) V2 CS OP
8) V2 CS CP
Split plot design:
V1
V2
CS
CP
OS
OS
OP
CS
Pest Control Treatments:
:
Organic (OP)
Conventional (CP)
 certified organic or untreated seed
 mechanical weeding
 refuge area to encourage beneficials
 bionet, collars, certified treatments
 chemically treated seed
 chemical weed control
 chemical pest control
e.g. garlic spray
Soil Treatments:
:
Organic (OS)
 4 year rotation (ley
Conventional (CS)
→ broccoli →
onion → carrot)
 additional organic fertilization as
 no set rotation (IPM)
 “other crop” in place of ley
 no winter cover crop
indicated by soil test *
 winter cover crop
*Based on soil test equivalent levels of N, P, K added to CS and OS treatments
Varieties:
:
Crop
V1
V2
Carrot
‘Flyaway’
‘Nairobi’
Broccoli
‘Fiesta’
‘Belstar’
Onion
‘Hyskin’
‘Red Baron’
Ley crop:
‘Other’crop:
red clover
Winter cover
crop:
Refuge:
rye + phacelia
wheat ‘Amaretto’ in 2009; lettuce ‘Design’ in 2010,
‘Design’ and ‘Tusca’ in 2011, Tonata in 2012
sunflower, borage, buckwheat, vetch, coriander,
fennel, cornflower, corn marigold, cocksfoot grass,
phacelia, nasturtium,
Factorial Systems Comparison Trial
Refuge 2009
Spring 2009
OS + OP
OS + CP
Harvested carrots
Lab Analyses:
AFRC
- Polyacetylenes in carrot
- Glucosinolates in broccoli
- Phenolic compounds in onion
Kinsealy
- Yield
- Total phenolics + flavonoids
- Taste panel test
- Soil ecology
Systems trial: Onion (2009 data)
Mean bulb weigh (g)
Onion - mean bulb weight (g)
100
Bulb weight was higher under CP
treatment (p<0.05)
75
50
25
0
OS + OP
CS + OP
OS + CP
CS + CP
Treatment
Onion - Total Phenolics
Total Phenolics – higher in ‘Red Baron’
(p<0.05) and under OS treatment
(p<0.01)
GAE (mg/100g FW)
500
400
300
200
100
0
OS + OP
CS + OP
OS + CP
CS + CP
Treatm ent
Onion - Total Flavonoids
Hyskin
CE (mg/100g FW)
60
Red Baron
50
Total Flavonoids – higher in ‘Red Baron’
(p<0.01) and under OS treatment
(p<0.01)
40
30
20
10
0
OS + OP
CS + OP
OS + CP
Treatment
CS + CP
Total Flavonoids higher under fully
organic (OSOP) than fully conventional
(CSCP) (p<0.05)
Systems trial – broccoli (2009 data)
Mean floret weight (g)
Broccoli - Mean floret weight (g)
300
Floret weight was higher under
CS treatment (p<0.01)
200
100
0
OS + OP
CS + OP
OS + CP
CS + CP
Treatment
GAE (mg/100g FW)
Broccoli - Total Phenolics
400
Total Phenolics – higher under
OS treatment (p<0.05)
300
200
100
0
OS + OP
CS + OP
OS + CP
CS + CP
Treatment
Broccoli - Total Flavonoids
Belstar
CE (mg/100g FW)
40
Fiesta
30
20
10
0
OS + OP
CS + OP
OS + CP
Treatm ent
CS + CP
Total Flavonoids – higher under
OS treatment (p<0.05)
Dr Juan Valverde AFRC
Glucosinolates (2009 and 2010 data)
2009
2010
Effect
NeoGBS
GBS
year
ns
ns
block
ns
ns
Var
ns
ns
Soil
ns
ns
Var*Soil
ns
ns
Pest
p<0.05
p<0.06
Var*Pest
p<0.05
ns
Soil*Pest
ns
ns
Var*Soil*Pest
ns
ns
OSOP vs CSCP
p<0.01
p<0.01
Systems trial - Carrot (2009 and 2010 data)
2009 - Total harvest weight
(kg per plot)
2010 - Total harvest weight
(kg per plot)
Nairobi
Nairobi
Flyaway
Flyaway
90
60
80
70
40
Yield (kg)
Yield (kg)
50
30
20
60
50
40
30
20
10
10
0
0
OS + OP
CS + OP
OS + CP
CS + CP
OS + OP
CS + OP
• Higher yield from cv. Nairobi both years (p<0.05)
• Higher yield with CS both years (p<0.01)
• Higher yield with CP treatment in 2009 (p<0.05)
OS + CP
CS + CP
Systems trial - Carrot (2009 and 2010 data)
• Total phenolics
• Total flavonoids
2010 - Total phenolics
2009 - Total phenolics
Nairobi
Flyaway
20
15
10
Flyaway
25
GAE(mg/100g FW)
GAE(mg/100g FW)
25
Nairobi
20
15
10
5
5
Flyaway CP)
0
0
OS + OP
CS + OP
OS + CP
OS + OP
CS + CP
2009 - Total flavonoids
CS + OP
OS + CP
2010 - Total flavonoids
Nairobi
12
CS + CP
Nairobi
Flyaway
12
Flyaway
10
CE (mg/100g FW)
CE (mg/100g FW)
10
8
6
4
2
8
6
4
2
0
0
OS + OP
CS + OP
OS + CP
CS + CP
Total Phenolics:
• Slightly lower in 2010
• No significant main effects
• Variety x pest control
interaction in 2009 (higher in
OS + OP
CS + OP
OS + CP
CS + CP
Total Flavonoids:
• Higher in 2010
• Significantly higher under
OS soil treatment in 2009
but not 2010
• Variety x pest control
interaction in both years
(higher in Flyaway CP)
Dr Juan Valverde AFRC
Polyacetylene content (2009 and 2010 data) - Falcarinol
Nairobi
Flyaway
250
200
150
100
50
0
OS+OP
OS+CP
CS+OP
2010 - FaOH content
FaOH content (ug/g DM)
FaOH content (ug/g DM)
2009 - FaOH content
CS+CP
Nairobi
Flyaway
250
200
150
100
50
0
OS+OP
OS+CP
• Significant effect of year (p<0.05)
• Falcarinol significantly higher in cv. Nairobi (p<0.01)
• Interaction effects…
•Overall conventional not different to organic
CS+OP
CS+CP
Soil Microbial Community
*in collaboration with Dora Lola-Luz, Bryan Griffiths, Dorothy Stone, Eileen Cullen
Biolog Eco Plates - CLPP
CSCP
OSOP
SUGAR DERIVATIVES
Well
SUGAR PHOSPHATES
Well
CARBOXYLIC ACIDS
Well
CLPP- Average well colour development
1
OSOP
OSCP
CSOP
0.8
CSCP
G1
H1
A2
B2
C2
D2
E2
H2
G2
B1
F2
A3
B3
C3
D3
E3
F3
G3
H3
AWCD
0.6
AMINO ACIDS
Well
0.4
0.2
A4
B4
C4
D4
E4
F4
G4
H4
2.82
0.74
1.91
0.54
-0.07
2.20
2.62
TOTAL SUGAR DERIVATIVES
10.75 ± 0.79
2
3
4
Day
5
POLYMERS
Well
Higher under OS p<0.001
Pest control treatment n.s.
Organic (OSOP) higher than conventional (CSCP)
C1
D1
E1
F1
f
4.97 ± 1.47
± 0.21
± 0.24
± 0.51
b
± 0.10
c
± 0.02
± 0.16
± 0.50
D,L--Glycerol Phosphate
Glucose-1-Phosphate
0.35 ± 0.03
0.09 ± 0.15
0.39 ± 0.06
-0.07 ± 0.07
TOTAL SUGAR PHOSPHATES
0.44 ± 0.17
0.31 ± 0.08
d
± 0.13
± 0.04
± 0.07
± 0.15
± 0.42
e
± 0.32
± 0.11
± 0.02
± 0.03
± 0.16
Pyruvic Acid Methyl Ester
D-Glucosaminic Acid
D-Galactonic Acid -Lactone
D-Galacturonic Acid
2-Hydroxy Benzoic Acid
4-Hydroxy Benzoic Acid
Hydroxybutyric Acid
Itaconic acid
Ketobutyric Acid
D-Malic Acid
1.96
0.29
1.80
1.90
0.60
2.36
0.15
1.19
-0.18
1.37
TOTAL CARBOXYLIC ACIDS
11.45 ± 0.46
L-Argenine
L-Asparagine
L-Phenylalanine
L-Serine
L-Threonine
Glycyl-L-Glutamic Acid
Phenylethylamine
Putrescine
TOTAL AMINO ACIDS
0
0.36
0.24
0.71
0.03
0.03
1.81
1.79
± 0.06
± 0.05
± 0.35
b
± 0.10
c
± 0.02
± 0.27
± 0.24
D-Cellobiose
-D-Lactose
-Methyl- D-Glucoside
D-Xylose
i-Erythritol
D-Mannitol
N-Acetyl-D-Glucosamine
a
a
2.03
3.75
0.49
2.37
0.16
0.25
1.46
1.90
g
0.64
1.86
0.03
0.92
± 0.06
± 0.13
± 0.12
± 0.58
TOTAL POLYMERS
5.73 ± 0.64
g
± 0.21
± 0.22
± 0.11
± 0.15
± 0.04
± 0.12
± 0.14
± 0.07
9.98 ± 0.42
12.40 ± 1.09
1.05
1.72
0.29
2.67
d
± 0.08
± 0.04
± 0.12
± 0.36
± 0.06
e
± 0.10
± 0.13
± 0.61
± 0.03
± 0.24
8.16 ± 1.15
1.48
3.25
0.16
2.13
-0.09
0.09
1.02
1.94
± 0.23
± 0.32
± 0.07
± 0.18
± 0.19
± 0.12
± 0.38
± 0.27
Tween 40
Tween 80
-Cyclodextrin
Glycogen
1.36
0.23
1.79
1.55
-0.01
1.43
0.20
1.22
-0.17
0.57
f
h
± 0.21
± 0.15
± 0.07
± 0.69
3.44 ± 0.87
h
Soil Microbial Community
Culturable Microbial and total nematode populations
Fungal/bacterial ratio
Soil Microbial Community
DGGE
Fungal primers:
Bacterial primers:
Band Identity
Closest hit
B1
Cellvibrio
spp.
B1c
Cellvibrio spp
Cellvibrio vulgaris
strain NCIMB 8633
(NR_025209.1)
Cellvibrio vulgaris
strain NCIMB 8633
(NR_025209.1)
B2a
E
Max identity
value
0.0
99%
0.0
99%
Probable
Nocardioides
spp.
NR_044981.2
2eNocardioides jensenii 108
strain KCTC 9134
97%
B3
Similar to
Deinococcus
spp.
Deinococcus aerius
strain TR0125
(NR_040934.1)
2e125
86%
B3b
Similar to
Geobacter
spp.
Geobacter
bemidjiensis Bem
strain
(NR_042769.1)
4e-53 93%
B2b
Excised and sequencing 22 fungal bands and 8 bacterial
Acknowledgements: Christy Roberts, Leo Finn, Philip Dunne, FETAC students
Thank you!