1. OPTIMIZATION OF FLY ASH FOR THE CULTURING OF AM FUNGI
Sanmit Adhikari1, Sayalee Dolas1, Neha Nair1, Pushkar Vartak1, Mili Awasthi1, Nikita Parab2, Seema Mishra2
1SIES- Graduate School of Technology
2SIES Indian Institute of Environment Management
Sri Chandrasekharendra Saraswati Vidyapuram
Nerul, Navi Mumbai 400 703
A.M. Fungi: a Ubiquitous Microorganism
 AM fungi are associated with at least 80% plants of the earth.
An obligate biotroph feeding only on the products of
photosynthesis
of their alive plant hosts
It consist of intra- and extraradical structures such as
arbuscules, vesicles,
and intraradical hyphae.
 The extraradical structures are extraradical hyphae,spores,
and auxiliary cells
PotentIal of A.M
FUNGI
Soil
Aggregate
formation
Reception of
direct supply
of C from host
Bio
remediation
Provision of
greater
absorptive
surface
Increases
Plant
Yield[2]
Provision
of greater
absorptive
surface
Culturing of AM Fungi
A.M. Fungi grows under axenic conditions and requires a
live host for growth.
Plants and grass family are known to be the best hosts for
their culturing.
A large quantity of soil is needed for the culturing of A.M.
Fungi
Requirement of AM fungi culture in bulk results in the loss
of upper fertile layer of soil.
Wastes viz. fly Ash can be used as a substitute for the
culturing of A.M. Fungi.
Fly Ash contains micro and macro nutrients as well as
some heavy metals.
AM fungi will help in the remediation of toxicity of heavy
metals present in fly ash as well as support their sustainable
culturing.
Objective of Study
To study the suitability of waste viz. fly ash as inoculum for
AM fungi.
To optimize the standard dose of fly ash for AM fungi
culturing.
Factors responsible for
phytostablization
and phytoextraction
Secretion of Chelating
agents.
Transport of heavy metal
in the hyphae of the
fungus.
The plasma membrane as
a living, selective barrier in
plants and fungi.
Sequestration of heavy
metal in the vacuole of
plant and fungal cells.
Metal Transfer in Plants
Role of AM fungi in heavy metal
remediation Gohre
and Paszkowski,2006
Materials and methods
1. Treatment Details:
Treatment Fly Ash
Concentration
Soil
Concentration
Treatment 1 0% 100%
Treatment 2 25% 75%
Treatment 3 50% 50%
Treatment 4 75% 25%
Treatment 5 100% 0%
2. Test Plant : Wheat
Procurement of AM fungi Culture: Glomus intraradiaces
from TERI, New Delhi
Physical Parameters Chemical Parameters
Electrical Conductivity
(EC)
(by Conductivity
Meter)
Macro Nutrients
(Organic Carbon,
Nitrogen (Kjeldahl
Method),
Phosphorous (Olsen’s
Method.)
pH
(by pH Meter)
Micro Nutrients (Lead,
Cadmium, Potassium,
Iron, Zinc, Magnesium,
Nickel, Manganese)
(by Atomic Absorption
Spectroscopy)
Water Holding
Capacity
4. Biometric Parameters [11]
Root Length
Plant Height
Fresh Weight
Dry Weight
5. Enumeration of A.M. Fungi
Spores(Philips and Hayman Method,
1970)
Results and Discussion
Parameter
s
Plant
Height
Root
Length
Fresh
Weight
Dry
Weight
Soil
(100%)
15.6±
2.306
2.93±
0.208
0.1±0.05 0.005 ±
0.010
Soil(75%)+
FA(25%)
25±
10.28
6.8±
2.9461
0.17±
0.14
0.023±
0.022
Soil(50%)+
FA(50%)
30.5±
3.3004
6.56±
1.34
0.273±
0.072
0.035±
0.015
Soil
(25%)+FA(7
5%)
29.1±
3.36
5.76±
1.15
0.236±
0.097
0.025±
0.01
FA(100%) 29.2±
2.55
8.51±
1.201
0.245±
0.042
0.03±
0.002
Parameters pH Electrical
Conductivity
(in mʊ)
Water
Holding
Capacity
(in %)
Organic
Carbon
Content
(in %)
Nitrogen
Content
(in ppm)
Phosph-
orous
Content
(in kg/ha)
Soil
(100%)
6.75 0.135±0.004 57.48 0.364 121.8 2.5088
Soil(75%)+FA(25%) 7.19 0.193±0.018 55.86 0.67 107.8 2.32
Soil(50%)+ FA(50%) 7.25 0.136±0.004 50.82 1.219 110.6 10.572
Soil (25%)+FA(75%) 7.72 0.167±0.004 50.5 1.121 77 3.6736
FA(100%) 8.78 0.266 51.49 0.518 19.6 7.52
Spore Density
0
500
1000
1500
2000
2500
Spore Density(per
100g of soil)
Root Infection
0
20
40
60
80
100
120
Root
Infection(%)
Atomic Absorption Spectroscopy for Cadmium
Concentration (mg/kg) Mean Standard
Deviation
Replicate
1
Replicate 2 Replicate 3
Pre-Soil 0.00085 -0.00127 -0.00338 -0.00183 -0.00136
Pre-Fly Ash 0.00126 0.00253 0.00042 0.00141 0.00106
Soil (100%) 0.00211 0.00254 0.00423 0.00296 0.00112
Soil (75%) +
Fly Ash (25%)
0.00507 0.00423 0.00423 0.00451 0.00049
Soil (50%)+Fly
Ash (50%)
0.00761 0.00423 0.00634 0.00606 0.00171
Soil (25%) +
Fly Ash
(100%)
0.00719 0.00507 0.00761 0.00662 0.00136
Fly Ash
(100%)
0.00761 0.00846 0.00888 0.00832 0.00065
Significant observations from study
Standard Dose of Fly Ash with Soil for culturing of
A.M. Fungi was found to be 50%.
50% fly ash with soil also showed the highest AM
fungi spore density (2295 spores per 100gm of soil)
along with optimal availability of organic carbon,
nitrogen and phosphorous content as compared to
treatment control treatment/
Maximum root infection was also observed in this
particular treatment (50% fly ash) confirms its
suitability for AM fungi culturing.
Scope of Future study
Field leveled trials for confirmation of results.
Similar study can also be done near ash ponds for
their management.
We express our gratitude to SIES IIEM, Nerul for
providing us the opportunity of conducting the
Project titled “OPTIMIZATION OF FLY ASH FOR THE
CULTURING OF AM FUNGI” in the final year, as per
the curriculum of the Degree course of
Biotechnology.
References
1. Marques AP, Oliveira RS, Samardjieva KA,
Pissarra J, Rangel AO, Castro PM. Solanum
nigrum grown in contaminated soil: effect of
arbuscular mycorrhizal fungi on zinc
accumulation and histolocalisation. 145(3), 2007
,pp 691-699
2. Satyawati Sharma , Seema Mishra.”Mycorrhiza-A
potencial approach for augmenting soil fertility
and productivity”.Nov 24, 2008, pp 87-114
Acknowledgements