ACADIAN SEAPLANTS LIMITED ACADIAN SEAPLANTS LIMITED ACADIAN SEAPLANTS LIMITED ACADIAN SEAPLANTS LIMITED ADVANCES IN THE USE OF ADVANCES IN THE USE OF ASCOPHYLLUM ASCOPHYLLUM NODOSUM NODOSUM SEAPLANT EXTRACTS FOR CROP SEAPLANT EXTRACTS FOR CROP PRODUCTION PRODUCTION PRODUCTION PRODUCTION LINKING LABORATORY AND FIELD RESEARCH LINKING LABORATORY AND FIELD RESEARCH Jeffrey Norrie, Ph.D., P.Ag. CPH Jeffrey Norrie, Ph.D., P.Ag. CPH Acadian Seaplants Ltd., Dartmouth, Nova Scotia, Canada Acadian Seaplants Ltd., Dartmouth, Nova Scotia, Canada
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� Gas Chromatography/Mass spectrometry� Specific molecules
� NMR technologies� Comparative profiles of molecular constituents
� In vivo testing
NMR Spectra of ASL ExtractsNMR Spectra of ASL ExtractsLocalities and SeasonsLocalities and Seasons
Ingomar NS Sep 2004
Barrington NS Jun 2004
Bear Point NS Mar 2004
Glenwood NS Jan 2004
Principal Components Analysis of Principal Components Analysis of 11HH--NMR SpectraNMR Spectra
12
14
A_14
A_15
6
8
10
G_1
H_1 H_2
I_4
0
2
4
PC2
A_1
A_2
A_3A_4A_5
A_6A_8
A_9A_10A_11A_12A_13 A_18A_19
A_20
A_21
A_22A_23A_24
A_25
A 26
A_27 A_28A_29A_30
A_31A_32 A_33A_34A_36
A 37A_38A_39A 40
A_41
A 42
A_43B_1
B_2C_1
D_1
F_1
I_1I_2
I_3
J_1
K_2
L_1L_2
-6
-4
-2_A_26 _A_35A_37A_40 A_42
E_2
E_3 K_1
-12
-10
-8 A_16A_17
E_1
-10 0 10 20PC1
HPLC Ultahydrogel Profile with PSA AssayAcadian SSEP 4318 (A1-04)
0.25
0.3
139.4
S
Very large 1,000kDa 500kDa 100kDa 50kDa 10kDa 1kDa
0.15
0.2so
rban
ce @
480
-490
nm
137.4
138.4
Inde
x O
utpu
t mV
PSA AssayRI Output
0.05
0.1
PSA
Assa
y, A
b
135.4
136.4
Refra
ctiv
e
015 17 20 22 25 27 30 32 35 37 40 42
Elution time, min
134.4
HPLC Ultahydrogel Profile with PSA AssayLSE Pressurized Cook
0.7
0.8
0.9
1
480-
490n
m
138.6
140.6
t mVPSA Assay
RI Output
Very large 1,000kDa 500kDa 100kDa 50kDa 10kDa 1kDa
0.3
0.4
0.5
0.6
A As
say,
Abs
orba
nce
@ 4
134.6
136.6
Refra
ctiv
e In
dex
Out
putp
0
0.1
0.2
15 18 20 23 25 28 30 33 35 38 40 43
Elution time, min
PSA
130.6
132.6
Points to Consider
� Different seaweed products are made in� Different seaweed products are made in different ways.Diff d h diff bi i i i� Different products have different bioactivities
� Need to identify these actives in various ymarine plant products
� Need to connect activities to specific plant� Need to connect activities to specific plant responsesGenetic/molecular responses� Genetic/molecular responses
Hypersensitive Vitamins
A ti id t
ypresponse
Vitamins
Hormones(stimulants /Anti-oxidants (stimulants /
retardants
PhenolicsOrganic
AcidsAmino acids
PolyaminesPhytoalexins Polyamines
Betaines SalicilatesJasmonatesGABA Jasmonates
Brassins
Stimuli
Stress ResistanceStress ResistanceS id di t i i b tS id di t i i b t
s/g
Superoxide dismutase in creeping bentgrassSuperoxide dismutase in creeping bentgrass
15
20
00 u
nits
5
10
ty (x
100
DM)
0
5
D ac
tivit
Low N High N
SOD
Treated Control
Virginia Polytechnical University, 1997Virginia Polytechnical University, 1997
Ascorbate peroxidase activity in Ascorbate peroxidase activity in tall fescuetall fescuetall fescuetall fescue
3500y
250030003500
se a
ctiv
ityw
t.)
100015002000
pero
xida
sol
e/h/
g fr.
0500
E+ E-scor
bate
(µ
mo
E+ E-As
Seaweed No Seaweed
Virginia Polytechnical UniversityVirginia Polytechnical University, 1997Virginia Polytechnical University, 1997
P/S capacity in creeping bentgrassP/S capacity in creeping bentgrassI h t th ti ffi i
0 390)
Improves photosynthetic efficiency
0.2
0.25
0.3ty
(Fvm
6
0.1
0.15
0.2
c ca
paci
t
0
0.05
0.1
synt
hetic
0Low N High N
Phot
o
Treated Control
Virginia Polytechnical University, 1997Virginia Polytechnical University, 1997
Resistance to Disease StressResistance to Disease Stress
University of Avignon, 1998
Resistance to Disease StressResistance to Disease Stress
University of Avignon, 1998
Seaweed extract:At least 2 applications‘Primer’ & ‘Activator’
Soil Applied
Increased
Greenerfoliage
Primer & Activatorroot & shootgrowth
(Fe & Laminarin + other microbe ‘food’)
P d l t
Feeds microbes
Produce plantgrowth stimulatingcompoundsRE
Alarm reactioninduced SAR
Direct induced SAR
(fluorescent Pseudomonads)
SiderophoreFe
ß-1,3-glucanase
induced SAR
Greater resistance Siderophore
production
Laminarin(Antagonism)
es sta ceto soilbornefungal diseases
ß-1,3-glucanases
(Fungal hyphae)Laminarin
(Antagonism)ß-1,3-glucans
Less disease (Antagonism)
Root Application Dec, 2002
Less diseasesymptomsOn plants (Also compete for space, nutrients, water & may produce antibiotics)
I d th & litI d th & lit•• Increased crop growth & quality:Increased crop growth & quality:¾¾ From plant establishment to harvestFrom plant establishment to harvest
•• Enhanced crop resistance to abiotic and biotic Enhanced crop resistance to abiotic and biotic stresses:stresses:¾¾ D ht li it h t hilli f t tD ht li it h t hilli f t t l i tl i t¾¾ Drought, salinity, heat, chilling, frost, waterDrought, salinity, heat, chilling, frost, water--logging, etclogging, etc¾¾ Disease and InsectsDisease and Insects
•• Provide commercially relevant technical Provide commercially relevant technical knowledge: knowledge: ¾¾ Market support for Market support for AscophyllumAscophyllum productsproducts..
Acadian Seaplants R&DAcadian Seaplants R&Dpp
•• Bioassay testing for bioactivityBioassay testing for bioactivity•• Processing developmentProcessing development•• Formulation of active fractionsFormulation of active fractions•• ShelfShelf--life studieslife studies•• Greenhouse efficacy testingGreenhouse efficacy testing•• Local replicated field trialsLocal replicated field trials•• Target market replicated field trialsTarget market replicated field trials•• Commercialization and release of informationCommercialization and release of information
MCSB: Product Testing
0.4a
Maize Chilling Stress Bioassay
ce (F
v/Fm
)0.3
a
abbbb
bhy
ll Flu
ores
cenc
0.2c
Chl
orop
h
0 0
0.1
0.0
Control (distilled water)SSEP Control (Batch 1186)ASL
ControlExtract 1
ASL Liquid Seaweed Concentrate 29%, pH 8 (May 17, 2005; Batch 5118)ASL Liquid Seaweed Concentrate 29%, pH 8 (May 20, 2005; Batch 5131)Beijing Leili (Spain, October 2004 (rec'd))Beijing Leili (USA, March 2005 (rec'd))Algifert (Norway, March 2005 (rec'd))
Figure 20. Bioactivity (Maize Chilling Stress Bioassay (MCSB)) of controls (applied at 100 mg/l or equivalent. Different letters indicate significantly different means between treatments (α = 5%). Error bars (fractions) represent standard error (SE). (n = 30)
Extract concentration
Figure 2. Effect of batch 5054 on lettuce hypocotyls elongation in the dark after 96 hours.
Extract concentration
Figure 1. WinRhizo root scanner and associated computer software analyze plant roots for important characteristics of root physiology such as root surface area, root diameter, root volume, p y gy , , ,and the number of root tips. .
Field Trials ProgramField Trials Programh fi ld i l i l j i i lh fi ld i l i l j i i lResearch field trials, coop trials, joint trialsResearch field trials, coop trials, joint trials
•• B ll 1 S iB ll 1 S i•• Ball 1: ScienceBall 1: Science¾¾How many times should I apply? How many times should I apply? ¾¾LittlLittl OftOft¾¾LittleLittle--OftenOften
•• Ball 2: EconomicsBall 2: Economics¾¾ H ti I ff d t l ?H ti I ff d t l ?¾¾ How many times can I afford to apply? How many times can I afford to apply? ¾¾ Consider ROIConsider ROI
•• Ball 3: PracticalBall 3: Practical•• Ball 3: PracticalBall 3: Practical¾¾ How many times will I actually apply?How many times will I actually apply?¾¾ Tank mixes, routines, weather, labourTank mixes, routines, weather, labour, , ,, , ,
ControlControlPlot: 3 x 10’Plot: 3 x 10’Crop: Processing tomatoesCrop: Processing tomatoes
AcadianAcadianAcadianAcadianPlot: 3 x 10’Plot: 3 x 10’Crop: Processing tomatoesCrop: Processing tomatoes
ControlControlPlot: 10’ rowPlot: 10’ rowCrop: Green PeppersCrop: Green PeppersRep: 3Rep: 3
TreatedTreatedPlot: 10’ rowPlot: 10’ rowCrop: Green PeppersCrop: Green PeppersRep: 2Rep: 2Rep: 3Rep: 3 Rep: 2Rep: 2
Chart 10: Acadian on Strawberries - Ventura County, Winter 2007 - Cumulative Marketable Production Net Return by Pick Day
Effect Of Acadian, Soil-Applied Post Veraison On Rachis Length In CRIMSON and RED GLOBE Grapes, Fresno, CA. 2002
ACADIAN
30
CONVENTIONAL
25
15
20
5
10
R hi T t l L th (CM) CRIMSON R hi T t l L th (CM) RED GLOBERachis Total Length (CM) CRIMSON Rachis Total Length (CM) RED GLOBE
Tobacco
A: TreatedB: Control
BEANS UNDER WATER STRESS
TREATEDCONTROL
Benefits of Benefits of AscophyllumAscophyllump yp yTake Home MessagesTake Home Messages
•• Benefits through foliar applications:Benefits through foliar applications:•• Improved plant nutrition, growth and Improved plant nutrition, growth and
d l td l tdevelopmentdevelopment
•• Improved fruit quality and quantityImproved fruit quality and quantity
•• Improved efficiency of crop inputsImproved efficiency of crop inputs
•• Enhanced disease insect and nematodeEnhanced disease insect and nematodeEnhanced disease, insect and nematode Enhanced disease, insect and nematode repressionrepression
Benefits of Benefits of AscophyllumAscophyllum
•• Benefits through soil applicationsBenefits through soil applications ::
p yp yTake Home MessagesTake Home Messages
Benefits through soil applicationsBenefits through soil applications ::¾¾Stimulate plant beneficial microbes and Stimulate plant beneficial microbes and
increase soil suppressiveness to diseasesincrease soil suppressiveness to diseasesincrease soil suppressiveness to diseasesincrease soil suppressiveness to diseases
¾¾ Induce a direct SAR effect via the rootsInduce a direct SAR effect via the roots
L d t b tt t bli h tL d t b tt t bli h t•• Leads to better crop establishment, Leads to better crop establishment, improved crop health and ultimately improved crop health and ultimately increased productivity plus profitability (to increased productivity plus profitability (to growers)growers)
Benefits of Benefits of AscophyllumAscophyllump yp yTake Home MessagesTake Home Messages
•• Ascophyllum nodosumAscophyllum nodosum extract is a extract is a general general biostimulantbiostimulant¾¾(vitamin(vitamin--like effects)like effects)¾¾Substantial research shows SAR benefitsSubstantial research shows SAR benefits
•• Can be used many times during the seasonCan be used many times during the season¾¾Foliar and/or soil applicationsFoliar and/or soil applications
•• Stimulates growth and development at time of Stimulates growth and development at time of applicationapplication
•• Can use often without toxicity problemsCan use often without toxicity problemsy py p
Benefits of Benefits of AscophyllumAscophyllume e ts oe e ts o scop y uscop y uTake Home MessagesTake Home Messages
•• Seaweed products can be made in different ways Seaweed products can be made in different ways (e.g. species, processing and extraction)(e.g. species, processing and extraction)¾¾May result in different activitiesMay result in different activities¾¾May result in different activities.May result in different activities.
•• Identifying active ingredients in Identifying active ingredients in AscophyllumAscophyllum extractsextracts¾¾FootFoot printing molecular profiles of our productsprinting molecular profiles of our products¾¾FootFoot--printing molecular profiles of our productsprinting molecular profiles of our products
•• Connecting activities with specific plant responsesConnecting activities with specific plant responses¾¾ In controlled bioassaysIn controlled bioassays¾¾ In controlled bioassaysIn controlled bioassays¾¾ In the fieldIn the field
•• Delivering bestDelivering best--use application information touse application information toDelivering bestDelivering best--use application information to use application information to growers and endgrowers and end--users on ongoing basisusers on ongoing basis
Stimulate proteins and growth promotinggrowth promoting