Microbial Biofertilizers andtheir Microbial Biofertilizers andtheir

Potential in sustainable
Agriculture
Dr. Heike Bcking
Outline
1. Overview Plant Microbe Interactions
2. Mycorrhizal interactions
ff f h l
4 I t ti b t h
3. Effect of mycorrhizal interactions on
nutrient uptake and pathogen resistance
4. Interactions between exchange processes
5. Nitrogen flux in the symbiosis
6. Mycorrhizal fungi and their application
Ectomycorrhiza Fagus
Overview Plant-Microbe Interactions
Bacteria
Nitrogen (N
2
) Fixation
Symbiotic bacteria (Rhizobia, Frankia)
Associated bacteria (Acetobacter - sugarcane)
Free living bacteria (Frankia rhizosphere) Free-living bacteria (Frankia rhizosphere)
Plant Growth promoting Bacteria
F i
Abrosimov 2007
Fungi
Mycorrhizal associations
Arbuscular mycorrhizal fungi y g
Ectomycorrhizal fungi
Other root colonizing fungi
Smith and Read 1997 Smith and Read 1997
Plant-Growth Promoting Bacteria
Enhanced N supply to the host by N
2
ppy y
2
fixation
Enhanced supply of other plant nutrients
( bl S d h l ) (P mobilization, S oxidation, Fe chelation)
Phytochrome production leading to
increases in root surface area (IAA,
Siberian soy beans, 1 control, 2 - nodule
bacteria; 3 - nodule bacteria and
pseudomonads (Dashkevish 2007)
c eases oot su ace a ea ( ,
cytokinin, gibberllin)
Enhancement of other beneficical
bacterial or fungal symbioses
The significance of mycorrhizal interactions
80-90% of all known plant species
bryophytes, pteridophytes,
gymnosperms and most of the
angiosperms angiosperms
simultaneous colonization by various
mycorrhizal types and fungal species,
essential for achlorophyllous plants
dependent on environmental conditions
I ncreasing attention for their role as
biofertilizers, bioprotectors and
bioregulators bioregulators
Smith and Read 1997
The benefits for both partners
Carbohydrates
Nutrients
Stress resistance
modified Egli, Brunner 2002
The life cycle and morphology of an
asexual coenocytic obligate symbiont
Germinating
spore
Spores
Appressorium
Spores
Sporulation
H h l b h
Arbuscule
Hyphal branches
Structure of an arbuscular mycorrhizal root
Mycorrhizal root (light microscope) Arbuscule (electron microscope)
How does the fungus-plant interaction work ?
I ncrease in the nutrient
absorbing surface area
beyond the depletion
zone of the root
Highly efficient nutrient Highly efficient nutrient
uptake systems
Better P storage
capabilities
Utilization of organic
nutrient resources
How does the fungus-plant interaction work ?
I ncreased nutrient supply
Competition among the microorganisms
for limited nutrient resources
Effect on the quantity and quality of
root exudates root exudates
Selective pressure on the microbial
populations in the rhizosphere leading to populations in the rhizosphere leading to
an increase in the number of
microorganisms with antagonistic
p ope ties properties
Activation of defense mechanisms by the
mutualistic fungus
Mycorrhizal and nonmycorrhizal barley
plants after colonisation with
Cochliobolus sativus (Kogel, Giessen)
mutualistic fungus
Nutrient exchange always includes an apoplastic step
Arbuscular interface
Intercellular interface
Arbuscular interface
Plant plasma membrane
Intercellular interface
Plant plasma membrane
Interfacial matrix
Plant cell wall
Interfacial matrix Interfacial matrix
Fungal cell wall
Interfacial matrix
Fungal cell wall
Fungal plasma membrane Fungal plasma membrane
arbuscular mycorrhiza arbuscular mycorrhiza
Nutrient exchange includes passive and active steps
Soil Soil Fungus Fungus Interfacial Interfacial
Apoplast Apoplast
Plant Plant
P
i
H
+
H
+
P
i
P
i
P
i
ATP
H
+
H
+
ATP
ADP
H
+
H
+
Hexose
acid
H
+
H
+
Hexose
Sucrose Sucrose
Invertase
H
+
ATP
H
+
H
+
ADP ADP
The C availability affects P uptake and P transfer
by arbuscular mycorrhizas
Sucrose Phosphate
C
C C C C
P
P Growth
Extraradical mycelium (ERM) Daucus root system
PolyP LP, DNA-P
PolyP LP, DNA-P P
i
Cytopl.
Bcking, Shachar-Hill, 2005
P uptake and P transfer is stimulated
by the carbohydrate availability
Soil Soil Fungus Fungus Interfacial Interfacial
Apoplast Apoplast
Plant Plant
P
i
H
+
H
+
P
i
P
i
P
i
ATP
H
+
H
+
P
i
H
+
H
+
P
i
P
i
P
i
ATP
H
+
H
+
P
i
H
+
H
+
P
i
P
i
Sinks P
i
ATP
H
+
H
+
ATP
ADP
H
+
H
+
ATP
ADP
H
+
H
+
Fungal
Carbohyd.
Growth
ATP
ADP
H
+
H
+
polyP
Hexose
acid
H
+
H
+
Hexose
H
+
H
+
Hexose Hexose
H
+
H
+
Hexose
Hexose-P
Hexose
acid
Sucrose Sucrose
Invertase
H
+
ATP
H
+
H
+
ATP
H
+
H
+
H
+
H
+
H
+
ATP
Photo-
synthesis
Sucrose Sucrose
Invertase
ADP ADP ADP ADP ADP ADP
Nitrogen flux in the arbuscular mycorrhizal symbiosis
Sucrose
Sucrose
NO
3
NH
4
Hexose
NO
3
NH
4
Hexose
Protein
C
Pool
C
Pool
ERM
Amino
acids
AA AA
IRM
ERM
INTERFACIAL
APOPLAST
SOIL
HOST
Nitrogen flux in the arbuscular mycorrhizal symbiosis
Sucrose
Sucrose
NO
3
NH
4
Hexose
NO
3
NH
4
NO
3
NH
4
Hexose
Protein
NO
3
NH
4
Protein
C
Pool
C
Pool
GS
ERM
Amino
acids
AA AA AA AA Amino
acids
IRM
ERM
INTERFACIAL
APOPLAST
SOIL
HOST
Nitrogen flux in the arbuscular mycorrhizal symbiosis
Sucrose
Sucrose
NO
3
NH
4
Hexose
NO
3
NH
4
Hexose
Protein Protein
C
Pool
C
Pool
ERM
Amino
acids
AA AA AA AA Amino
acids
IRM
ERM
INTERFACIAL
APOPLAST
Arg-N
SOIL
HOST
Nitrogen flux in the arbuscular mycorrhizal symbiosis
Sucrose
Sucrose
NO
3
NH
4
Hexose
NO
3
NH
4
Hexose
Protein
C
Pool
C
Pool
C
Pool
ERM
Amino
acids
AA AA AA Amino
acids
IRM
ERM
INTERFACIAL
APOPLAST
Arg-C
SOIL
HOST
Nitrogen flux in the arbuscular mycorrhizal symbiosis
Sucrose
Sucrose
NO
3
NH
4
Hexose
NO
3
NH
4
Hexose
Glu
Arg
Gln
C
Pool
NH
4
NH
4
Orn
Urea
NH
4
Arg
C
Pool
ERM
PolyP
P
PolyP
P
i
P
i
P
i
Arg
IRM
ERM
INTERFACIAL
APOPLAST
P
i
i i i
SOIL
P
i
HOST
The dependency of various crop species on mycorrhiza
Mycorrhiza Potential yield loss Crops
dependency without mycorrhiza
Very high Greater than 90 % Linseed
High 60 80 % Sunflower, mungbean, g , g ,
pigeon pea, maize,
chickpea
Medium 40 60 % Sorghum, soybean
Low 10 30 % Wheat, barley, triticale
Very low 0 10 % Panicum, canary
Nil 0 Canola lupins
The State of Queensland (Department of Primary Industries and Fisheries)
Nil 0 Canola, lupins
Application of mycorrhizal fungi
Arbuscular mycorrhizal fungi can account for 5 50 % of the Arbuscular mycorrhizal fungi can account for 5 50 % of the
microbial biomass in the soil (Ryan and Graham, 2002)
Efficient mycorrhizal symbiosis can substitute 222 kg P
2
O
5
ha
-1
2 5
(Kelly et al., 2001)
I n field studies, the growth yield of linseed could be correlated
to the mycorrhizal colonization rate (Thompson et al 1991) to the mycorrhizal colonization rate (Thompson et al., 1991)
High P levels in the soil reduce significantly the colonization
rate.
Why do we need mycorrhizal research?
We need to better understand:
Regulation of transport processes (beneficial nutrient transport Regulation of transport processes (beneficial nutrient transport
in relation to the carbon costs for the plant)
Complex role of arbuscular mycorrhizal fungi in resource
allocation
Partner choice in mycorrhizal systems
Effects of mycorrhizal colonization on the nutritional value Effects of mycorrhizal colonization on the nutritional value
I nteractions between arbuscular mycorrhizal fungi and root
pathogen under field conditions p g
P management
Studying metabolism and transport in
arbuscular and ectomycorrhizal associations
Soil
culture
In vivo
Sample
analysis
0
1
10
2 4 8 24 48 96
Molecular
tools
Localization
and regulation
Monoxenic culture
ERM ERM Ac Plant Ac
Autoradiography
GC/MS
LSMand analytical
Monoxenic culture
Data analysis
LSMand analytical
electron microscopy
In
vitro
Axenic culture
Data analysis
vitro
Labeling Labeling with stable with stable
or radioactive or radioactive
isotopes isotopes
Sustainable
agriculture
Revegetation
Remediation
Research Questions
Nitrogen metabolism and transport in the symbiosis
Sucrose
Sucrose
NO
3
NH
4
Hexose
3 3
3
Gln
C
NO
3
NH
4
Hexose
NH
4
NH
4
Orn
Urea
NH
4
Glu
C
1
1
3
PolyP
Arg
C
Pool
P P P
Urea
Arg
C
Pool
PolyP
1 2
2
IRM
ERM
INTERFACIAL
APOPLAST
P
i
P
P
i
P
i
P
i
SOIL
P
i
HOST
Activity and regulation of the urea cycle in the IRM
1
Interactions between the N and P flux in the symbiosis 2
Gene expression, in situ hybridization, labeling studies, GC/MS, microautoradiography,
enzymatic assays, Western blots, EDXS
Regulation of plant and fungal N transporters and interactions of C and N flux
3
The application of mycorrhizal fungi
in sustainable agriculture
Maximal benefit of mycorrhizal fungi
by:
I noculation with efficient mycorrhizal fungi
I ncrease of activity by proper cultural y y p p
practices
Cultural practices that increase the Cultural practices that increase the
activity
Reduced tillage
Crop rotations
Cover crops
Phosphorus management p g
Collaborators
Yair Shachar- Hill, Michigan State University
Philip E. Pfeffer, USDA Wyndmoor
Peter Lammers, New Mexico State University
Toby Kiers, University Amsterdam
Arbuscule
Thanks for your attention