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Introduction to endophytes and their application to develop commercial products

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Ponencia: Introduction to endophytes and their application to develop commercial products …

Ponencia: Introduction to endophytes and their application to develop commercial products

Autor: Dr. Gary Strobel

Evento TF Innova: Workshop Biotechnology "Isolation and identification of endophytic fungi from vascular plants"


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Transcript

  • 1. Montana State University The Promise of Endophyte Biology
  • 2. The Causal Agent of Dutch Elm Disease – Ceratocystis ulmi
  • 3. European Bark Beetle
  • 4. Antibiosis Effects of the Pseudomycins
  • 5. Transformation Plasmid
  • 6. The Field Experiment
  • 7. Systemic Candida albicans Infection in a Child
  • 8. Control of Black Sigatoka with Pseudomycin B
  • 9. Taxomyces andreanae
  • 10. Taxomyces andreanae an Endophyte on Yew
  • 11. Thin Layer Chromatography of Fungal Taxol and Authentic Taxol
  • 12. Electrospray MS of Fungal Taxol
  • 13. Ipecac Products from the Rainforest Taxol Vinblastine Reserpine Tubocurarine
  • 14. Definition of An Endophyte By definition an endophyte is a microorganism that becomes established in a higher plant and does not cause any overt symptoms or cause any apparent tissue damage
  • 15. An Endophyte from the Jurassic Period
  • 16. Areas of High Plant Biodiversity 70-80% of the Earth’s Plant Diversity Occurs on 1.5% of Its Land Area Is this also true of microbial diversity?
  • 17. ant Diversity in the Canary Isles 0 million year old landscape.. About 2000 plant species About 528 of these are endemic Chances of finding novel endophytes are great
  • 18. Selecting Plants for the Isolation of Endophytes 1. Visiting Areas of High Plant Biodiversity 2. Finding Gondwanaland Plants 3.Meeting and Learning from Native Peoples
  • 19. Progress on Endophytic Microbes Examples of products-jesterone, hydroxyjesterone pestacin, isopestacin, ambuic acid, ecomycin, oocydin A munumbicins, kakadumycin, cryptocandin, pseudomycins and others. Ambuic Acid: JOC (2003) 4609-4614
  • 20. A Bastion of Biodiversity-the World’s Rainforests
  • 21. Avenue of the giants
  • 22. The Spiny Forest of Southern Madagascar
  • 23. Angel Falls in the Tepui Lands of Venezuela Highest in the world- 3212’
  • 24. Top of the Roraima Tepui
  • 25. A Novel Fungus Seimatoantlerium tepuiense
  • 26. Igauazu Falls-Brazil/Argentina
  • 27. An area of ancient Gondwanaland-Torres Del Paine, Chile
  • 28. Technique of Isolating Endophytes
  • 29. Some Endophytic fungi
  • 30. Tripterigeum wilfordii a Chinese medicinal plant used to treat arthritis and other ailments of the joints and limbs possesses a plethora of endophytic microbes including Cryptosporiopsis quercina.
  • 31. Cryptosporiopsis quercina
  • 32. Cryptosporiopsis spores
  • 33. HPLC Separation of Cryptocandin using an Altima C-18column Bioactivity
  • 34. Cryptocandin MW 1079 Pneumocandin MW 1079
  • 35. Biological Activity of Cryptocandin Test Fungal MIC MIC of Pathogen micrograms/ml Amphotericin B of Cryptocandin Candida albicans 0.03 0.03 Cryptococcus > 20.0 0.04 neoformans Aspergillus >20.0 0.03 fumigatus Histoplasma 0.01 0.01 capsulatum
  • 36. Crystals of Cryptocin
  • 37. X-ray Crystallographic Device
  • 38. Structure of Cryptocin
  • 39. Carrao River
  • 40. Rhyncholacis penicillata under a water fall on the Carrao River
  • 41. Serratia marcescens on the plant surface
  • 42. Serratia marcescens Activity Against Pythium
  • 43. The Papuans
  • 44. Pestalotiopsis jesterii, a fungal endophyte
  • 45. Conidiospores of P. jesterii
  • 46. Ambuic Acid is produced by various isolates of Pestalotiopsis microspora obtained in various rainforest locations around the world
  • 47. Monstera speciosa in the Jungles of the Upper Amazon of Peru
  • 48. P-25 An Endophytic Streptomycete from the Monstera vine
  • 49. Origin of Spores in P-25 vs. Penicillum sp.
  • 50. Some Characteristics of Isolate P-25 1. Is endophytic on Monstera speciosa 2. Appears to be a streptomycete a. Only bacterial primers function to give product from its ribosomal DNA b. Primers for 18Sr DNA do not function c. Has no discernable nucleus (by nuclear staining) d. Gram positive 3. Has 16S r DNA sequences that have 95% similarity to Streptoverticillum sp. 4. Makes Coronamycins 4. Has some characteristics of Penicillum sp. but origin of spores is unique 5. Is not inhibited by cyclohexamide up to (or greater) than 100 ppm
  • 51. Activity of P-25 Against Pythium
  • 52. Anopheles sp. vector of Plasmodium sp.- cause of Malaria
  • 53. Properties of Coronamycins 1.Peptides- 2 principle ones MW.’s 1217 and 1203 2. Composition x:x methionine:tyrosine and leucine-ratio-- 2:2:1:1:3 3. Active against pythicaeous fungi (2-16 micrograms/ml) ,and Cryptococcus neoformans. 4. Extremely active against Plasmodium falciparum IC 50 <1 ng /ml
  • 54. My Native Australian Friends
  • 55. Northern Territory Australia – a Monsoonal Rainforest Snakevine-Kennedia nigriscans
  • 56. Streptomyces sp. A-11 from Snakevine
  • 57. HPLC Separation of the Components of Streptomyces sp. A-11 Munumbicins E-4,5 M.W. 1315 & 1431 Actinomycin D Actinomycin X2 Actinomycin X0 beta
  • 58. Some Representative Streptomycetes from Snakevine
  • 59. New, Very Unusual Streptomycetes spp. from Snakevine
  • 60. Streptomycete 149 -- Horn Structure
  • 61. Rainforest Microbiology- The Prospects and Promises 1. Each individual plant has one or more associated microorganisms. Only a few microbes have ever been studied. 2. The relationships of the plant to the microbe may range from symbiotic to weakly pathogenic. 3. Host specificity may exist with certain microbes which places a huge emphasis on the preservation of the world’s rainforests. 4. Products from endophytes have a wide range of uses in medicine, agriculture and industry. 5. Peculiar microbes are plentiful and usually are to be found in areas of high biodiversity. 6. The biology of endophytes is largely unknown.