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 seminar - i

Antagonistic and Synergistic Interaction in

Microbial Consortium and their Influence
on Plant Growth

Archana, D .S.
PAK 8014
 Introduction

Microbial communities showed varied

forms of interaction ranges from synergistic and
mutualistic to antagonistic and parasitic.

Microbial interaction depends on biotic and abiot

ic components of the environment helps in
transformation, mobilization and solubilization
of essential plant nutrients.
 Types of interactions in rhizosphere
1. Interaction between plant roots and microorganisms

2. Interaction between microorganisms

 Ecological Associations Among
Microorganisms
Symbiotic – organisms live in close nutritional relationships;
required by one or both members
 Mutualism – obligatory, dependent; both members benefit
 Commensalism – commensal member benefits, other
member not harmed
 Parasitism – parasite is dependent and benefits; host is
harmed

Non-symbiotic – organisms are free-living; relationships

not required for survival
 Synergism – members cooperate and share
nutrients
 Antagonism – some member are inhibited or
5
destroyed by others
 Criteria to be considered while designing
Microbial consortium

“Interactions among the microorganisms”

“Negative interaction” “Positive interaction”

Antibiosis Neutralism
Mutualism
Competition
Synergism
Amensalism
Symbiosis
Predation Protocooperation
Parasitism Commensalisms
 Effect of Interaction
Population/ Population/
Type of Interactions Species A Species B
Neutralism 0 0

Commensalism 0 +

Mutualism + +

Amensalism 0 -

Predation, parasitism + -

Competition - -
INTERACTION EFFECT
 Antagonistic interactions
“Any inhibitory effect of an organism created by any
means to the other organism (S) is antagonistic
interaction”

 Used in biological control of plant pathogens

 Antagonist are not pathogen specific
 Inhibits wide range of microorganisms
 True parasitic relationship with microbial host
 Mechanisms are not mutually exclusive
Mechanisms of Antagonism

Amensalism : Inhibition or destruction of one organism by a

metabolite produced by another organism.

Eg. Antibiotics, volatile compounds, enzymes etc.

Antagonistic activity of T.viride on

Bortytis cinerea, Cladosporium sp

(Bai et al, 2008)

 Microorganisms produce different type
Microorganisms Antibiotics
Antibiotics
Target pathogen Disease

Pseudomonas 2, 4- Pythium spp. Damping off

fluorescens F113 diacetylphloroglucino
l
Agrobacterium Agrocin 84 Agrobacterium Crown gall
radiobacter tumefaciens
Trichoderma Gliotoxin Rhizoctonia solani Root rots
virens

Bacillus subtilis Bacillomycin D Aspergillus flavus Aflatoxin

contamination

Burkholderia Pyrrolnitrin,pseudane R. solani and Damping off

cepacia Pyricularia oryzae and rice blast

P. fluorescens Phenazines Gaeumannomycs Take-all

2-79 and 30-84 graminis var. tritici
 Competition

Competition: two or more organisms trying to

utilize the same nutrients or occupy the same
niche or infection site.
 Parasitism
Predation or Parasitism : Attack and
feed directly on the target organism
or the biocontrol agent can produce
some sort of toxin that kills the target
organism and then the biocontrol
agent feeds on the dead target.

Parasitism
1. Hyperparasites – are parasites of
parasites

2. Mycoparasites - fungi that parasitizes

T. Harzianum coils around
other fungi Rhizoctonia Solani

(Benhamou and Chet, 2003)

 Antagonisms leading to biological control of plant
pathogens
Type Mechanism Examples
Lytic/some nonlytic mycoviruses
Direct antagonism Hyperparasitism/predation
Ampelomyces quisqualis
Pasteuria penetrans
Trichoderma virens

Mixed-path Antibiotics 2,4-diacetylphloroglucinol

Phenazines
antagonism Cyclic lipopeptides
Chitinases
Lytic enzymes
Glucanases
Proteases

Unregulated waste products Ammonia

Carbon dioxide
Hydrogen cyanide
Blockage of soil pores
Physical/chemical interference
Germination signals consumption
Molecular cross-talk confused
Exudates/leachates consumption
Indirect antagonism Competition
Siderophore scavenging
Physical niche occupation
Contact with fungal cell walls
Induction of host resistance
Detection of pathogen-associated,
molecular patterns
Phytohormone-mediated induction
 Synergistic Interaction of Beneficial
Microorganisms
(PGPR, Rhizobium and Arbuscular Mycorrhizal
fungi)

Microorganisms in combinations interact synergistically

 Providing nutrients
 Removing inhibitory products
 Stimulating each other
 Enhance beneficial aspects of their physiology.
 Soil microbiological equilibrium
 Favorable environment for plant growth
 Mechanisms of interaction in
beneficial microorganisms
Direct mechanisms:
1. Phytohormone
2. P-solubilization
3. Nitrogen fixation
4. Induce systemic resistance
5. Decreased heavy metal toxicity

Indirect mechanism:
1. Siderophores
2. Anti-fungal metabolites -antibiotics,
3. Fungal cell wall-lysing enzymes,
4. Hydrogen cyanide,
5. Competition for ‘rhizosphere space’ and nutrients
6. parasitism
 Microbe-Microbe interaction

1. PGPR and Rhizobium

2. AM fungi and rhizosphere microbes

 PGPR and Rhizobia interact synergistically for N2
fixation
Mechanisms of action:

 Altering the host secondary metabolism or creating antibiosis

 Stimulate formation of additional infection sites- Auxins

 Alteration of the plant flavonoid metabolism

 Promoted the nod-gene inducers in roots

 By stimulate the plant to produce more signal molecules

 Phytohormones- Siderophores ,phytoalexins,and flavonoids

 Decreased heavy metal toxicity

 Increase in soil enzyme activities

Synergistic interactions between PGPR and Rhizobia on
plant growth

PGPR strain Rhizobium strain Benefit as an

increase in
Pseudomonas sp Bradyrhizobium sp N2 and P uptake in
green gram
Pseudomonas sp Rhizobium Shoot height, root
leguminosarum length and dry
biovar viceae weight of pea
Pseudomonas striata Rhizobium sp Pigeonpea
Azospirillum sp
Pseudomonas B.japonicum Soyabean
fluorescens
 AM fungi and rhizosphere microbes

AM fungi interact natural and introduced

microorganism ‘mycorrhizosphere’

Rhizobacteria acted as ‘Mycorrhiza-Helper-Bacteria’

(MHB) produces biologically active substances

 Stimulate mycelial growth and germination

 Mycorrhizal colonization
 Spore germination
 AM fungi Interact synergistically to stimulate plant growth

 Improved nutrient acquisition

 Inhibition of fungal plant pathogen
 Enhancement of root branching
 Physical attachment results - carbon uptake by the bacteria from the
fungal hyphal surface by provide them with a competitive advantage.

Direct interactions
 Supply of energy-rich C compounds derived from host plants
 Changes of mycorrhizosphere i.e pH induced by the fungus
 Competition for nutrients
 Fungal exudation of inhibitory or stimulatory compounds

Indirect interactions
 Modification of root exudates and soil structure
Groups of bacteria interact with AM
fungi

1. AMF with symbiotic nitrogen fixers

2. AMF with Asymbiotic nitrogen fixers
3. AMF with phosphate solubilizing bacteria
4. AMF with PGPR
 AMF AND RHIZOBIA

Interact synergistically and their effects on plant growth:

 AM fungi improve P uptake - energy available for N2 fixation

 AMF enhances the functioning of Nitrogenase

 Improve nodulation and N2 fixation under low water potential

 Uptake micronutrients by the AM fungi –improve plant growth,

indirect effects upon the N2-fixing system

 AM fungi increased nutrient status -mycorrhizosphere, by

decomposing organic N compounds, benefit additional nitrogen
provided through N2 fixation
AMF with Asymbiotic nitrogen fixers
AMF – affects population of other microorganisms in the
rhizosphere both quantitatively and qualitatively

Ex: A. chroococcum and Glomus fasculatum in tomato rhizoplane

 Increased population of A. chroococcum in rhizosphere for

long time
 Enhanced AM infection and spore production
 Increased N2 fixation and P avaliability
Effect of specific AM sp on population of bacteria
Ex: P. fluroscence decreases after infection of cucumber seedling
with G. intraradices not after G. etunicatum
Azospirillum - polysaccharide-degrading bacteria
C source for N2 fixation

Azospirillum – Bacillus
Increased N2 fixation

Azospirillum brasilense - Enterobacter cloacae and

Arthrobacter giacomelloi -efficient N2 fixation

Azospirillum sp. DN64 - Cellulolytic fungi

nitrogenase activity increased

Azospirillum brasilense –Staphylococcus

Increased N2 fixation due to release of aspartic acid
 AMF with phosphate solubilizer
Synergistic interactions results
 Bacteria promoted Mycorrhizal pre colonization
establishment
 Survival PSB longer around Mycorrhizal roots

Effect on plant growth

 PSB solubilizing more Phosphorous
 Mycorrhiza enhance P uptake
 Increased plant biomass and dry matter production
 N and P content in plant tissues- plant hormones or
vitamins than P solubilization
 AMF with PGPR
PGPR
1. Improved mineral nutrition
2. Soil fertility and plant health
3. Disease suppression,
4. Phytohormone production

PGPR interact synergistically with AM fungi

 Mycorrhizal establishment and function

 Improves mycelial growth of AM fungi
 Stimulation of root development
 Helps in recognition process between root and fungus.
 Enhanced AM fungal colonization levels in roots
 Gram-positive bacteria and γ-proteobacteria interact
synergistically with AM fungi
 Synergistic interactions between bacteria and AM fungi, to
enhanced plant growth.
Bacterial species AMF species Effect
Gram+, low G+C Glomus clarum ↑f.g.,↑s.g.,↑r.c.
Bacillus pabuli G. intraradices ↑p.s.,↑r.c.
Bacillus subtilis G. intraradices ↑f.g.,
Paenibacillus validus G. mosseae ↑f.g.,↑s.g.,↑r.c.+i.p.p.f
Paenibacillus sp.
Gram+, high G+C G. versiforme ↑s.g. ↑ s.g.
Corynebacterium sp. Gigaspora margarita
Streptomyces orientalis
Y-Proteobacteria G. Intraradices ↑p.s.,↑r.c.
Enterobacter sp. G. versiforme ↑s.g.
Pseudomonas sp Endogone sp. ↑f.g.,↑r.c.
Pseudomonas aeruginosa G. Intraradices ↑p.s.
Pseudomonas putida G. Intraradices ↑r.c.
Rhizobium meliloti G. mosseae ↑Nitrogen fixation
rates
Antagonistic interactions between bacteria and
AM fungi

 Inoculation of G. fasciculatum and Streptomyces

cinnamomonicus with finger millet inoculation had
antagonistic effect-

 Suppressed the growth and multiplication of the other

 Microbial competition and suppressed colonization-result

failure establishment of AMF

Krishna et al., 1982

 Microbial consortium

“Group of different species of microorganisms

that act together as a community”.

 Organisms work together in a complex system where all

benefit from the activities of others in the community.
 Microbial consortia are much more efficient than single
strains of organisms with a diversity of metabolic capabilities.
Ideal Characteristics of microbial consortium

 Fast acting and high rhizosphere competence

 Synergistic to each other
 Able grow with or without air
 Produces natural enzymes - Wide degrading ability
 Easy to handle and mass multiplication
 Broad spectrum of action
 Long shelf life and good stability
 Should tolerate desiccation, heat, oxidizing
and UV radiations
 Non-toxic, non-pathogenic and non-corrosive
 Economical and safe to environment
 Advantages
 Single strains of microorganisms are not capable of degrading
all of the compounds, therefore microbial consortia are essential
in the complete mineralization of any compounds.

 A microbial consortium is more resistant to environmental

shock.

 Compete and survive in the environment than single

microorganisms.

 Microbial consortia are capable of handling a wide variety

of complex wastes.
 “ Two-species microbial consortium for growth
promotion of Cajanus cajan”
(Pandey and Maheshwari, 2006)

Studied the interactions and the importance of plant

growth promoting consortium
Burkholderia sp. MSSP and Sinorhizobium meliloti PP3.
Indole-3-acetic acid (IAA)
Solubilize inorganic phosphate.
Nitrogen fixer
The organisms were grown as monospecies or mixed-
species culture/ consortium
Studied for growth profile, IAA production and phosphate
solubilization.
Fig 1: Effect of mixed-species consortium (U) on phosphate solubilization
compared to monospecies culture of Burkholderia sp. MSSP and S. meliloti PP3.
(Pandey and Maheshwari, 2006)
Fig 2: Effect of mixed-species consortium (U) on IAA production
compared to monospecies culture of Burkholderia sp. MSSPG and S.
meliloti PP3.
(Pandey and Maheshwari,
Table 1: Effect of Burkholderia sp. MSSP and Sinorhizobium
meliloti PP3 on seed germination and vegetative growth of
Cajanus cajana after 40 days of seed germination

Growth measurement
Sl Strain Seed Total Plant Shoot Fresh root Fresh shoot
No germination length length (mm) weight (mg) weight
(%) (mm) (mg)
1 Control 70 125 d 101 c 401 d 2.15 c
2 MSSP 100 188 b 150 b 835 a 3.18 b
3 PP3 90 142 c 111 c 560 c 3.47 b
4 MSSP+ 100 312 a 242 a 750 b 6.06 a
PP3

(Pandey and Maheshwari, 2006)

“Dual inoculation of Azotobacter chroococcum and
Glomus fasciculatum improves growth and yield
of sunflower under field condition”
(Sreeramulu et al., 2000)

 Field experiment
 Consisted nine treatments
 Sunflower (KBSH-1)
 Seeds are treated with the consortium
containing Azotobacter chroococcum and
Glomus fasciculatum
Table 2: Effect of dual inoculation of Azotobacter
chroococcum and Glomus fasciculatum on growth
and yield of sunflower at different levels of fertilizers
Sl. Treatments Plant No. of Head Yield
No. Height leaves diameter (q/ha)
(cm) (cm)
1 NPK (50 %) 143 18 11.8 10.00
2 NPK (75 %) 140 20 13.0 11.00
3 NPK (100 %) 145 20 15.0 15.50
4 NPK 50 % + Azotobacter 144 20 12.5 12.00
5 NPK 75 % + Azotobacter 145 19 13.5 13.20
6 NPK 50 % + Glomus fasciculatum 146 20 14.6 14.50
7 NPK 75 % + Glomus fasciculatum 148 20 15.2 15.80
8 NPK 50 % + Azotobacter + Glomus 150 21 17.6 16.20
fasciculatum
9 NPK 75 % + Azotobacter+ Glomus 154 22 19.4 18.50
fasciculatum
SEm ± 2.19 NS 0.90 1.20
CD (0.05) 6.58 2.70 3.59
CV (%) 2.60 10.59 14.72

(Sreeramulu, et al.,2000)
“ Studies on synergism between Rhizobium, plant
growth promoting rhizobacteria (PGPR) and phosphate
solubilizing bacteria in blackgram”
(Gunasekaran et al., 2000)
Field trial (National Pulse Research Center, Vamban)
Test crop: Balckgram
1. Uninoculated control
1. Rhizobium (COC. 10)
1. PSB (Bacillus megaterium)
2. PGPR (Pseudomonas KB 133)
3. Rhizobium + PSB
4. Rhizobium + PGPR
5. PGPR + PSB
6. Rhizobium + PSB + PGPR
RBD
Table 3: Synergisms between Rhizobium, PGPR and PSB

Sl. Treatments Plant No. of Plant Grain % increase

No height nodules biomass yield over
(cm/pl) /plant (g/pl) (kg/ha) control
1 UIC 32.4 10 13.3 472 -
2 Rhizobium (COC 10) 29.5 14 19.3 655 38.7
3 PSB (B. megaterium) 31.9 13 18.3 483 2.3
4 PGPR 29.3 14 28.3 616 30.5
(Pseudomonas KB 133)
5 Rhizobium + PSB 29.8 13 25.0 612 29.6
6 Rhizobium + PGPR 30.9 14 15.0 702 48.7
7 PGPR + PSB 31.8 12 25.0 483 2.3
8 Rhizobium + PSB + 37.3 19 28.3 760 61.0
PGPR
CD (P=0.05) 2.3 14 2.3 36.6

(Gunasekaran, et al., 2007)

 “Developing ‘ Microbial Consortia’ for better
growth and nutrition of Dalbergia sissoo”
(Raghu et al., 2005)
 D. sissoo – Avenue tree and legume
 Interaction between Glomus fasiculatum, PGPRs viz.,
Azotobacter chroococcum, Bacillus coagulans and
Trichoderma harzianum
Preparation of Microbial Consortium
 Azotobacter chroococcum, Trichoderma harzianum
and Bacillus coagulans
 Grown in separate medium. Mycelia mat of
Trichoderma harzianum and bacterial cultures was
macerated and centrifuged at 5000 rpm for 7 minutes
and the pellet was suspended with 0.1 M MgSO4
 10 ml of culture was added to the pots
 Table 4: Influence of Glomus fasiculatum and PGPRs
on the plant parameters of Dalbergia sissoo
Sl. Treatments Plant height Total Plant P Plant N
No. (cm) biomass (g) content content
(mg/pl) (mg/pl)
1 Control 62.1 c 10.6 c 19.7 abc 4.7 e
2 Glomus 66.0 bc 11.3 bc 19.7 abc 8.1 b
fasciculatum(Gf)
3 Azotobacter 69.7 abc 12.0 bc 29.6 c 5.2 de
chroococcum(Ac)
4 Bacillus coagulans(Bc) 65.9 bc 10.7 c 23.2 abc 5.3 de

5 Trichoderma harzianum 68.9 abc 10.7 c 21.1 cd 6.4 cd

(Th)
6 Gf+Ac 68.2 abc 11.7 bc 15.8 cd 5.5 d
7 Gf+Bc 64.6 bc 11.9 bc 26.6 abc 6.6 c
8 Gf+Th 67.0 bc 12.6 bc 27.3 c 6.4 cd
9 Gf+Ac+Bc 81.2 a 14.8 a 59.9 a 9.1 a
10 Gf+Ac+Th 62.2 c 11.7 bc 18.4 bcd 7.6 bcd
11 Gf+Ac+Bc+Th 61.8 c 12.7 bc 42.2 b 8.2 b
(Raghu et al., 2005)
 Synergistic effects of plant-growth promoting
rhizobacteria and Rhizobiumon nodulation and nitrogen
fixation by pigeonpea (Cajanus cajan)

TILAK et al., 2006

 Table 5: Effect of PGPRs and Rhizobium on plant biomass and
grain yield of pigeon pea cv.p-921
Treatments Dry plant biomass/g plant Grain yield/g plant

Uninoculated control 3.5 1.05

Rhizobium alone 4.2 1.25

Rhizobium+ Azotobacter 4.0 1.30

chroococcum
Rhizobium+ Azospirillum 4.0 1.35
brasilense
Rhizobium + Pseudomonas 4.2 1.75
fluorescens
Rhizobium+ Pseudomonas 4.8 1.75
putida
Rhizobium + Bacillus 4.0 1.62
cereus
Standard error 0.47 0.36

TILAK et al., 2006

 Table 6 :Nodulation ,nitrogen fixation and total nitrogen(N) content in
shoot of pigeonpea after inoculation with rhizobium and five PGPRs

Treatments Nodule Nodule dry Nitrogen Shoot N %

density/nodule/ weight fixation/mm
plant /g/plant ol C2H4
Uninoculated control 4.50 15.20 5.50 1.75

Rhizobium alone 12.50 30.50 8.90 1.78

Rhizobium+ Azotobacter 15.70 30.50 10.60 1.65

chroococcum
Rhizobium+ Azospirillum 15.20 38.50 11.20 1.78
brasilense
Rhizobium + Pseudomonas 22.00 55.80 12.90 1.90
fluorescens
Rhizobium+ Pseudomonas 27.50 57.00 13.80 2.05
putida
Rhizobium + Bacillus cereus 21.70 45.00 11.90 2.00

Standard error 3.19 4.19 0.55 0.05

 Impact of bio inoculants consortium on rice root
exudates biological nitrogen fixation and plant growth
(Raja et al 2006)
Table 7: Biochemical analysis of root exudates

(Raja et al 2006)
Table 8: Estimation of plant growth promoting substances in root
exudates
(Raja et al 2006)
 Table 8: Plant biometric observations
Treatments Root Shoot Dry weight Chlorophyll
length(c length(cm) (mg/plant) content
m) (mg)
T1-A.lipoferum 12.25 15.50 630 2.09
T2-P.fluorescens 8.20 15.30 580 2.03
T3-B.megaterium 4.30 6.10 227 0.63

T4- 20.30 16.30 720 2.30

Microbialconsortium(T1
+T2+T3)

T5-Un inoculated control 3.70 4.60 185 0.45

SED 2.28 2.96 127.66 0.2492

CD (0.05) 5.25 6.83 294.40 0.5748

(Raja et al ., 2006)
Response of Chilli(Capsicum annum) to inoculation
with Glomus mosseae,Pseudomonas fluorescences
and Azospirillum brasilense

Muthuraju and Jayasheela (2005)

 Table 9: Effect of AM fungi and PGPR on growth and yield of Capsicum
annum
Treatments Plant height Mean root Dry weight (g/plant) Fresh fruit
(cm) length (cm) weight (g/plant)
shoot root Total
biomass
Uninoculated control 65.6e 7.6e 0.94f 0.36f 1.3f 276.7e

Azospirillum brasilense 87.3c 9.5d 1.3e 0.47e 1.7e 357.1d

Pseudomonas fluorescens 77.3d 9.8d 1.4e 0.47e 1.9e 375.8c

Glomus mosseae 90.0b 11.0c 1.7bc 0.47c 2.2c 405.3ab

Azospirillum 91.0ab 12.8ab 1.5d 0.52d 2.0d 399.7b

brasilense+P.fluorescens

Glomus mosseae+A.brasilense 90.0b 12.5ab 1.7bc 0.58c 2.3c 405.4ab

Glomus mosseae+P.fluorescens 91.5ab 12.0b 1.8b 0.62b 2.5b 416.3ab

Glomus mosseae+P.fluorescens+ 92.5a 13.0a 2.1a 0.72a 2.8a 419.8a

A. brasilense

Means with the same superscript do not differ significantly at p=0.05

 Table 10: Effect of AM fungi and PGPRs on nitrogen, phosphorus content and
mycorrizal Colonization and spore count in root zone soil of Capsicum annum
Treatments Nitrogen (mg/plant) phosphorus Colonization Spore
(%) No./25g
shoot root Shoot Root soil

Uninoculated control 17.6e 4.0f 1.2e 0.31h 40.3f 138.0f

Azospirillum brasilense 22.4d 5.3c 1.7d 0.38g 50.0e 162.3f

Pseudomonas fluorescens 23.3d 5.7e 1.9bc 0.42f 47.3e 169.0e

Glomus mosseae 27.2c 7.2c 2.2a 0.46d 71.6b 172.3d

Azospirillum brasilense+P.fluorescens 23.4c 6.7d 2.1 0.48de 55.6d 165.0f

Glomus mosseae+A.brasilense 27.4c 7.4c 1.9bc 0.51cd 67.3c 182.3b

Glomus mosseae+P.fluorescens 30.4b 8.2b 2.2a 0.60b 72.0b 176.0c

Glomus mosseae+P.fluorescens+ A. 38.5a 9.6a 2.2a 0.64a 79.3a 190.3a

brasilense

Means with the same superscript do not differ significantly at p=0.05

 With increasing concerns regarding the impact
of conventional fertilizers and pesticides, use of
microbial consortium appear praised for a greater
role in future because these open up a world
opportunity and create living spaces which
ultimately bring about harmony in nature without
affecting the ecosystem.
Title of research

Development and Evaluation of

Microbial Consortium for Plant
Growth Promotion
 Objectives of investigation

1. Development of alginate based consortium of

microorganisms (Azotobacter, Pseudomonas sp and
Acinetobacter).

2. To test the survival of microbial consortium in alginate

formulation.

3. To study the release of microorganisms from alginate beads.

4. To determine effectiveness of selected formulation under

pot culture study.
THANK YOU