development and commercialization for bio pesticide by using pseudomonas

DEVELOPMENT AND COMMERCIALIZATION FOR BIO-PESTICIDE BY USING PSEUDOMONAS

FLUORESCENCE

INTRODUCTION P. fluorescence is gram negative bacillus

These bacteria are easy to culture, as they grow on minimal media and at 25-30 degrees Celsius

They destroy certain toxins and pollutants including styrene, polycyclic aromatic hydrocarbons, and TNT

They also protect plants from infection by pathogens by producing secondary metabolites like antibiotics and hydrogen cyanide which kill other bacteria and fungi

ABSTRACT In physical characteristic test in various media P. fluorescence shows positive growth

In biochemical characteristic on various media P. fluorescence shows positive result

In anti-pathogenic activity test of P. fluorescence also shows positive result against F. oxysporum. This test confirm that P. fluorescence can be use as bio-fertilizer

MATERIAL AND METHOD All the pure culture of P. fluorescence are taken from DESIGN

BIOSIS

All chemical & media were prepared to evaluate the biochemical assay, physical appearance & anti-pathogenic activity test of P. fluorescence

Also microscopic observation of P. fluorescence was done

Physical activity test

various media was used to determine the physical appearance of P. fluorescenceThose are King’s B agar media King’s Broth media Starch agar and starch broth media LB Broth media pikovaskaya's agar and broth media

Biochemical activity test

various media was use to determine the Biochemical activity test for P. fluorescenceThose are Citrate agar slant media Starch agar media Czapek agar medium

Anti-pathogenic activity test

various media was use to determine the anti-pathogenic activity test for P. fluorescenceThose are LB agar media Potato dextrose agar medium

Physical characteristic photos

King’s B agar medium

King’s B Broth media LB Broth media

Starch Broth media Starch agar media

Pikovaskaya's agar media Pikovaskaya's agar media

Biochemical activity test photos

Citrate agar medium

Starch agar medium

Czapek agar medium

Anti-pathogenic activity testIn PDA media F.oxysporum

P. fluorescence

Mat based inhibition of F.oxysporium(Plant Pathogen)

In PDA media

F. oxysporum P. fluorescence F.oxysporum

Inoculum based inhibition of F.oxysporium (Plant -Pathogen)

In LB agar mediaF. oxysporum

P. fluorescence

F. oxysporium is inoculated in middle and P. fluorescence Inoculated in around that F. oxysporium(Plant pathogen)

In LB agar media

F. oxysporum

P. fluorescence

Mat based inhibition of F.oxysporum (Plant Pathogen)

Mass Culture, Bio-formulation and commercialization

Blander

Mass culture ofPseudomonas fluorescence

Market product

RESULTPhysical activity test

From the above photos we can observed that in starch agar media the growth of Pseudomonas fluorescence is greater than Kings B media, pikovaskaya's media.

In fact in pikovaskaya's agar slant media was to less.

Biochemical activity test The biochemical assay confirmed that Pseudomonas

fluorescence has citrate utilizing, starch hydrolysis and cellulose activity.

This experiment shows the proof and practical identification

and confirmation method for P. fluorescence.

Anti-pathogenic activity test The anti-pathogenic activity of P. fluorescence against F.

oxysporum shows that the mat based formulation of bio-fertilizer is effective against F. oxysporum rather than liquid based preparation.

Also P. fluorescence can be added in combination with other soil friendly microorganisms to inhibit of the plant pathogens including F. oxysporum in it’s soil based application

Mass Culture, Bio-formulation and commercialization

The bio-formulation of P. fluorescence is an easy way if can be done by P. fluorescence Mat mixing with Talc powder and CMC.

The commercialization of this bio-formulation can be widely used as the plant pathogen eradication or control by using natural resources are very few in Agriculture sector.

REFERENCE www.biotecharticle.com/agriculture/stages in plant disease

development www.promusa.org/fussarium wilt, fussarium wilt of banana Agrios, George N. Plant Pathology. 3rd ed. New York:

Academic Press, 1972. print. Martinelli, F., Scalenghe, R., Davino, S., Panno, S., Scuderi,

G., Ruisi, P., Villa, P., Stroppiana, D., Boschetti, M., Goulart, L.R., Davis, C.E., Dandekar, A.M. "Advanced methods of plant disease detection. A review". Agronomy for Sustainable Development(2014).

DR. A. V. Navarajan Paul. Indian Agricultural Research Institute. Insect Pests and their Management.2007

G.N Agrious,plant pathology(3rd edition).journal of basic microbiology.1989

King E.O,Ward MK and Roney D.E.Lab and clinic.44.301-307(1954)

Harrigan W and Mecance M, Laboratory method of food and diary Microbiology.Academic press inc(London)ltd.(1976)

Andre K. Gonsalves. Department of Plant Pathology, University of Hawaii at Manoa

Guy R. Knudsen and Louise-Marie C. Dandurand. Ecological Complexity and the Success of Fungal Biological Control Agents. 2014,

LoekasSoesanto, EndangMugiastuti, and Ruth FetiRahayuniati. MORPHOLOGICAL AND PHYSIOLOGICAL FEATURES OF Pseudomonas fluorescence P60.2011

Pablo I. Nikel,EstebanMartínez-García&Víctor de Lorenzo. Ecological habitats of Pseudomonas spp.

Biotechnological domestication of pseudomonads using synthetic biology.368-379.2014

Girja Ganesh& Manoj kumar. Journal of plant interaction. Pseudomonas fluorescence,a potential

bacterial analogist to control plant disease;2005,123-13 AshahMhana Mohammad Laith K.T ALani,LassatBekbaya and

Baharauddinsalleh.World applied scinceJournal.Biocontrol effect of Fussarium oxysporium f .spaubense by Pseudomonas fluorescence and BABA invitro.2011.189-191

John stavrinides.Plant Pathogenic bacteria.Origin and evolution of pathogenic Bacteria.37,70-331(2009)

Jackson Rt.W. Plant Pathogenic Bacteria,Genomics and Molecular Biology.37(2009)

Creamer,Rebecca,H.Hubble,A.Lewis.PlantDisease,Curtovirus infection of chilli paper in maxico.480-486(May2005)

Avsveg.com.au/intranet/technical-insight/crop protection/bacterial disease.htm

R.K Mishra,R.KJaswal ,D.Kumar,PR Sable ,a.singh.Plant Breeding and Crop Science Management of Major disease and insect pest of onion and garlic.6(11).160-170(November2014)

Eaton A.D cleseriL.S,Rice E.W and Greenberg A.W. standard Methods for the examination of water and waste water.(2005)

Palleroni, N.J. Pseudomonadaceae. Bergey's Manual of Systematic Bacteriology. Krieg, N. R. and Holt J. G. (editors) Baltimore: The Williams and Wilkins Co., pg. 141 - 199(1984)

Anzai; Kim, H; Park, JY; Wakabayashi, H; Oyaizu, H; et al. (Jul 2000). "Phylogenetic affiliation of the pseudomonads based on 16S rRNA sequence". Int J Syst Evol Microbiol50 (4)1563–89

Rao, Subba. Soil Microbiology. Fourth ed. Enfield: Science Publishers, 1999.

Vrieze, Jop de (2015-08-14).Science 349 (6249)

THANK U!

ANYQUESTIONS!