OPTIMIZATION OF FLY ASH FOR THE CULTURING OF AM FUNGISanmit Adhikari1, Sayalee Dolas1, Neha Nair1, Pushkar Vartak1, Mili Awasthi1, Nikita Parab2, Seema Mishra2

1SIES- Graduate School of Technology2SIES Indian Institute of Environment Management

Sri Chandrasekharendra Saraswati VidyapuramNerul, 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 hostsIt 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 FungiA.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 StudyTo 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 methods1. 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 : WheatProcurement of AM fungi Culture: Glomus intraradiaces from TERI, New Delhi

Physical Parameters Chemical ParametersElectrical 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 LengthPlant HeightFresh WeightDry Weight5. Enumeration of A.M. Fungi Spores(Philips and Hayman Method, 1970)

Results and DiscussionParameters Plant

HeightRoot 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(75%)

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 ElectricalConductivity(in mʊ)

WaterHoldingCapacity(in %)

Organic Carbon Content(in %)

NitrogenContent(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

Treatment 1

Treatment 2

Treatment 3

Treatment 4

Treatment 5

0

500

1000

1500

2000

2500

Spore Density(per 100g of soil)

Root Infection

0

20

40

60

80

100

120

Root In-fection(%)

Atomic Absorption Spectroscopy for CadmiumConcentration (mg/kg) Mean Standard

Deviation

Replicate 1

Replicate 2 Replicate 3

Pre-Soil 0.00085 -0.00127 -0.00338 -0.00183 -0.00136Pre-Fly Ash 0.00126 0.00253 0.00042 0.00141 0.00106Soil (100%) 0.00211 0.00254 0.00423 0.00296 0.00112Soil (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 studyStandard 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.

References1. 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