Control of Rhizoctonia solani species and rootknot nematodes using chitinolytic and
antibiotic producing Pseudomonas speci...
Structure of Presentation
 Introduction of the problem
 Methods
 Application

 Benefits of Proposed Controls
 Conclus...
Introduction
R. solani disease symptoms
oLeaf Spot
oStem & root rot
oDamping-off
oSore shin

Nematode disease symptoms
oRo...
Identification of specific Pseudomonas species
Suppressive soils e.g. P. fluorescens, P.
chlororaphis, & P. aeruginosa
II....
Production of Antibiotics Using the ProEasyTM BEVS Technology
e

Antibiotic encoding gene
Polyhedrin promoter
Linearized P...
Production of Inoculum
 Batch culture within Kings-B Broth

 24-36 hours to harvest
 Cell densities high hence high ino...
Application of Controls
 Bacterial suspension & antibiotic solutions as
root dip & drenches
 Soil inoculants at transpla...
Broad spectral action of controls
Broad spectral action of controls continued…
Benefits of Technology
 Promotes healthy plant growth via nutrient uptake,

root growth & induced systemic resistance (IS...
Conclusion
 Efficient control of Rhizoctonia solani and root-

knot nematode species demands an integrated
management app...
Recommendations
Consider application of Pseudomonas species
inoculants & antibiotic products as broad
spectrum controls
II...
Acknowledgements
 Dr Garwe

 Mr. Muzhinji
 Plant Health Clinic (PHC) Personnel
 MBS, PB & PHC Students
THANK YOU
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Control of rhizoctonia_solani__species_and_root_knot_nematodes_using_chtinolytic_and_antibiotic_producing_pseudomonas_species

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A methed for controlling Rhizoctonia solani species and root-knot nematodes using chitinolytic and antibiotic producing Pseudomonas species

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Control of rhizoctonia_solani__species_and_root_knot_nematodes_using_chtinolytic_and_antibiotic_producing_pseudomonas_species

  1. 1. Control of Rhizoctonia solani species and rootknot nematodes using chitinolytic and antibiotic producing Pseudomonas species BY EUGENE MADZOKERE MOLECULAR BIOLOGY SERVICES CHINHOYI UNIVERSITY OF TECHNOLOGY
  2. 2. Structure of Presentation  Introduction of the problem  Methods  Application  Benefits of Proposed Controls  Conclusion & Recommendations  Acknowledgements
  3. 3. Introduction R. solani disease symptoms oLeaf Spot oStem & root rot oDamping-off oSore shin Nematode disease symptoms oRoot galls oStunted growth oChlorosis
  4. 4. Identification of specific Pseudomonas species 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 I.
  5. 5. Production of Antibiotics Using the ProEasyTM BEVS Technology e Antibiotic encoding gene Polyhedrin promoter Linearized ProEasy Tm Baculovirus DNA 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
  6. 6. Production of Inoculum  Batch culture within Kings-B Broth  24-36 hours to harvest  Cell densities high hence high inoculum yield
  7. 7. 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
  8. 8. Broad spectral action of controls
  9. 9. Broad spectral action of controls continued…
  10. 10. 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
  11. 11. 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
  12. 12. Recommendations Consider application of Pseudomonas species inoculants & antibiotic products as broad spectrum controls II. Adopt alternative screening techniques for Chitinolytic activity within antagonistic soilborne microflora III. Produce and commercialize the 3 antibiotics & Pseudomonas Inoculants I.
  13. 13. Acknowledgements  Dr Garwe  Mr. Muzhinji  Plant Health Clinic (PHC) Personnel  MBS, PB & PHC Students
  14. 14. THANK YOU

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