isolation and characterisation of ldpe degrading bacteria having plant growth promoting actives

8/22/2019 Isolation and Characterisation of LDPE Degrading Bacteria Having Plant Growth Promoting Actives

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Project Proposal (Primary Protocol)

Isolaon and characterisaon of LDPE degrading

bacteria having plant growth promong acvesAabeer Kumar Basu (Microbiology Department, Vijaygarh Jyosh Ray College)

Anirban Dua (Chemistry Department, Krishnagar Government College)

Key-Words:Low Density Polythene (LDPE), LDPE degradaon, N-xaon, P-solubilisaon, IAA-producon

Objecve:This project is aimed at isolaon, characterisaon and culvaon of Low Density Poly Ethene

(LDPE) degrading bacteria and assessing their plant growth promong acvies. For assaying

plant growth promong abilies, three properes will be studied: ability to x nitrogen,

ability to solubilise inorganic phosphorous and producon of Indole Acec Acid (IAA). Aer

being through these tasks, an aempt would be made to manufacture a bio-ferliser using

the bacterial strains isolated in this project.

Methodology:

(The tests/experiments and their procedures, as on requirement, may be changed.)1. Making LDPE powder: LDPE lms would be cut into small pieces, immersed in xylene

and boiled for 15 minutes and then crushed. LDPE powder so obtained would be washed

with ethanol to remove residual xylene and allowed to evaporate to remove excess

ethanol. Powder would be dried in hot air oven at 600C overnight (refer to no. 5 and 6 in

the reference list).2. Collecon of samples: Soil samples will be collected from various waste disposal vats

and dustbins. If possible, with permission, soil samples will be collected from

_________________________________________________________.3. Isolaon and characterisaon of LDPE degrading bacterial strains: Selecve enrichment

techniques would be employed to isolate LDPE degrading bacteria. The media used will

be minimal media with LDPE as chief carbon source (refer to no. 8 in the reference list).

Characterisaon will be done through various staining techniques and biochemical tests

(refer to no. 8 in the reference list).4. Assessment of LDPE degradaon: LDPE degradaon by the bacterial strains would be

monitored using any one or a combinaon of the following methods:


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4.1.By measuring the change in tensile strength and percentage extension of the LDPElms aer being inoculated with bacteria for a me period (refer to no. 6 in the

reference list).4.2.By measuring the Biological Oxygen Demand (BOD) of the media with LDPE as sole

carbon source (refer to no. 6 in the reference list).4.3.By trubidimetric analysis of the media, and plong the growth curve of the bacterial

strains (refer to no. 6 in the reference list).4.4.Microscopic analysis of LDPE lms aer being inoculated with bacteria for a me

period (refer to no. 4 in the reference list).4.5.By using modied Sturm Test, i.e., quanfying the amount of CO 2 produced by the

bacteria by degrading LDPE (refer to no. 4 in the reference list).5. Assessing N-xing abilies of the isolated strains: N-xing ability of isolated bacterial

strains would be assessed using either or both the following methods:5.1.By assessing their acetylene reducon acvity (ARA), using Burks N-free medium.5.2.By using N-free mannitol agar plate.

6. Assessing P-solubilising abilies of the isolated strains: P-solubilisaon test would beconducted qualitavely by plang the isolated bacterial strains in agar containing

precipitated tricalcium phosphate. Bacterial culture would be streaked on the surface of

the agar plates. The presence of clearing zone, if present, around the bacterial colonies

aer overnight incubaon would be considered indicaon of P-solubilisaon.7. Assessing IAA producon by the isolated strains: Paen and Glick (refer to no. 2 and 7

in the reference list) method of assaying IAA producon would be used.8. Producing bio-ferliser: This secon is provisional and completely dependent on the

success of the previous secons. Aer isolang LDPE degrading bacteria having the threebefore menoned plant growth promong acvies, a trial would be made to check

their abilies to enhance plant growth in combinaon with each other. The chance of

their success when used as bio-ferliser will be observed.

References:1. Husen, E., Screening of soil bacteria for plant growth promoon acvies in vitro;

Indonesian Journal of Agricultural Science 4(1) 2003:27-31.2. Paen, C.L., and Glick, B.R., Role of Pseudomonas puda Indole Acec Acid in

Development of Host Plant Root System; Appl. Environ. Microbiol. 2002, 68(8):3795.3. Karnwal, K., Producon of IAA by uorescent Pseudomonas in the Presence of L-

Tryptophan and Rice Roots Exudates; Journal of Plant Pathology, 2009, 91 (1), 61-63.4. R. Pramila and K. Vijay Ramesh, Biodegradaon of LDPE by Fungi Isolated From

Marine Water a SEM Analysis; African Journal of Microbiology Research, Vol. 5(28),

pp. 5013-5018.5. Kumar, S., et al, Isolaon and Idencaon of LDPE Degrading Fungi from Municipal

Solid Waste; JCPRC, 2013, 5(3):78-81.


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6. Sharma, A., and Sharma, A., Degradaon Assessment of Low Density Polythene (LDP)and Polythene (PP) by an Indigenous Isolate of Pseudomonas stutzeri; JSIR, Vol. 63,

March 2004, pp 293-296.7. Shakina J. et al, Microbial Degradaon of Synthec Polyesters form Renewable

Resources; Indian Journal of Science, Volume 1, Number 1, November 2012.8. Cappuccino, J. G., and Sherman, N., Microbiology: A Laboratory Manual; 7 th edion,

Pearson Educaon, Inc., Pages: 53-92, 203-208, 459-466, 512.

isolation and characterisation of ldpe degrading bacteria having plant growth promoting actives

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