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![Page 1: Control of rhizoctonia_solani__species_and_root_knot_nematodes_using_chtinolytic_and_antibiotic_producing_pseudomonas_species](https://reader033.fdocuments.us/reader033/viewer/2022052506/557d2905d8b42a7a608b4b18/html5/thumbnails/1.jpg)
BY
EUGENE MADZOKERE MOLECULAR BIOLOGY SERVICES CHINHOYI UNIVERSITY OF TECHNOLOGY
Control of Rhizoctonia solani species and root-knot nematodes using chitinolytic and antibiotic producing Pseudomonas species
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Structure of Presentation
Introduction of the problem
Methods
Application
Benefits of Proposed Controls
Conclusion & Recommendations
Acknowledgements
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Introduction
R. solani disease symptoms oLeaf Spot oStem & root rot oDamping-off oSore shin
Nematode disease symptoms oRoot galls oStunted growth oChlorosis
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Identification of specific Pseudomonas species
I. Suppressive soils e.g. P. fluorescens, P. chlororaphis, & P. aeruginosa
II. Morphological: Colloidal Chitin test
III. Spectral: fluorescence at 365nm
IV. Molecular:
a. PCR with pseudomonas specific primers :
i. PsF/PsR
b. PCR with antibiotic specific primers :
i. PRN - pyrrolnitrin
ii. Phl2a/Phl2b - 2, 4-diacetylphloroglucinol &
iii. PCA - phenazine-1-carboxylic acid
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Polyhedrin promoter
Plasmid transfer vector e.g. PVL 1393/ PAcAB3
Co-transfection into expressSF+ cells
Transfer expressSF+ cells for fermentation in Serum-free medium
Scale-up of fermentation for harvesting high antibiotic yield
Filtration & harvesting of cell-free extract
Ethyl acetate extraction of Antibiotics
Detection & Purification of Antibiotics by Chromatography
e
Production of Antibiotics Using the ProEasyTM BEVS Technology
Antibiotic encoding gene
Linearized ProEasy Tm
Baculovirus DNA
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Production of Inoculum
Batch culture within Kings-B Broth
24-36 hours to harvest
Cell densities high hence high inoculum yield
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Application of Controls
Bacterial suspension & antibiotic solutions as root dip & drenches
Soil inoculants at transplanting & after
Low cost carriers e.g. peat, calcined clay, powdered corn cobs, alginate microbeads
50-94% pathogen inhibition efficacy reported against AG 2-2, AG-3, AG-4, AG-5 R. solani species and Meloidogyne incognita species
Cameroon, USA, Netherlands, India, Australia, South Korea, China, Canada successfully controlled pathogenic species
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Broad spectral action of controls
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Broad spectral action of controls continued…
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Benefits of Technology
Promotes healthy plant growth via nutrient uptake, root growth & induced systemic resistance (ISR)
High quality antibiotic yield
Production processes highly scalable
High recombinant viral titer & cell densities
Control is non-phytopathogenic, environmentally safe & cost efficient.
Applicable with tobacco, rice, wheat, bean, potato, & citrus fruits
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Conclusion
Efficient control of Rhizoctonia solani and root-knot nematode species demands an integrated management approach.
Making use of
i. High efficacy fungicides, nematicides, biocontrols & prudent agric-practices
ii. Environmentally safe agrochemical controls
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Recommendations
I. Consider application of Pseudomonas species inoculants & antibiotic products as broad spectrum controls
II. Adopt alternative screening techniques for Chitinolytic activity within antagonistic soil-borne microflora
III. Produce and commercialize the 3 antibiotics & Pseudomonas Inoculants
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Acknowledgements
Dr Garwe
Mr. Muzhinji
Plant Health Clinic (PHC) Personnel
MBS, PB & PHC Students
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THANK YOU