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

Autor: Dr. Gary Strobel

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

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Introduction to endophytes and their application to develop commercial products

  1. 1. Montana State University The Promise of Endophyte Biology
  2. 2. The Causal Agent of Dutch Elm Disease – Ceratocystis ulmi
  3. 3. European Bark Beetle
  4. 4. Antibiosis Effects of the Pseudomycins
  5. 5. Transformation Plasmid
  6. 6. The Field Experiment
  7. 7. Systemic Candida albicans Infection in a Child
  8. 8. Control of Black Sigatoka with Pseudomycin B
  9. 9. Taxomyces andreanae
  10. 10. Taxomyces andreanae an Endophyte on Yew
  11. 11. Thin Layer Chromatography of Fungal Taxol and Authentic Taxol
  12. 12. Electrospray MS of Fungal Taxol
  13. 13. Ipecac Products from the Rainforest Taxol Vinblastine Reserpine Tubocurarine
  14. 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. 15. An Endophyte from the Jurassic Period
  16. 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. 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. 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. 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. 20. A Bastion of Biodiversity-the World’s Rainforests
  21. 21. Avenue of the giants
  22. 22. The Spiny Forest of Southern Madagascar
  23. 23. Angel Falls in the Tepui Lands of Venezuela Highest in the world- 3212’
  24. 24. Top of the Roraima Tepui
  25. 25. A Novel Fungus Seimatoantlerium tepuiense
  26. 26. Igauazu Falls-Brazil/Argentina
  27. 27. An area of ancient Gondwanaland-Torres Del Paine, Chile
  28. 28. Technique of Isolating Endophytes
  29. 29. Some Endophytic fungi
  30. 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. 31. Cryptosporiopsis quercina
  32. 32. Cryptosporiopsis spores
  33. 33. HPLC Separation of Cryptocandin using an Altima C-18column Bioactivity
  34. 34. Cryptocandin MW 1079 Pneumocandin MW 1079
  35. 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. 36. Crystals of Cryptocin
  37. 37. X-ray Crystallographic Device
  38. 38. Structure of Cryptocin
  39. 39. Carrao River
  40. 40. Rhyncholacis penicillata under a water fall on the Carrao River
  41. 41. Serratia marcescens on the plant surface
  42. 42. Serratia marcescens Activity Against Pythium
  43. 43. The Papuans
  44. 44. Pestalotiopsis jesterii, a fungal endophyte
  45. 45. Conidiospores of P. jesterii
  46. 46. Ambuic Acid is produced by various isolates of Pestalotiopsis microspora obtained in various rainforest locations around the world
  47. 47. Monstera speciosa in the Jungles of the Upper Amazon of Peru
  48. 48. P-25 An Endophytic Streptomycete from the Monstera vine
  49. 49. Origin of Spores in P-25 vs. Penicillum sp.
  50. 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. 51. Activity of P-25 Against Pythium
  52. 52. Anopheles sp. vector of Plasmodium sp.- cause of Malaria
  53. 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. 54. My Native Australian Friends
  55. 55. Northern Territory Australia – a Monsoonal Rainforest Snakevine-Kennedia nigriscans
  56. 56. Streptomyces sp. A-11 from Snakevine
  57. 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. 58. Some Representative Streptomycetes from Snakevine
  59. 59. New, Very Unusual Streptomycetes spp. from Snakevine
  60. 60. Streptomycete 149 -- Horn Structure
  61. 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.

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