Se ha denunciado esta presentación.
Utilizamos tu perfil de LinkedIn y tus datos de actividad para personalizar los anuncios y mostrarte publicidad más relevante. Puedes cambiar tus preferencias de publicidad en cualquier momento.

Instructions for the collection of plant specimens and endophytes studies

1.073 visualizaciones

Publicado el

Ponencia: Instructions for the collection of plant specimens and endophytes studies

Autor: Dr. Gary Strobel

Evento TF Innova:

Workshop Biotechnology "Isolation and identification of endophytic fungi from vascular plants"

  • Inicia sesión para ver los comentarios

  • Sé el primero en recomendar esto

Instructions for the collection of plant specimens and endophytes studies

  1. 1. The story of Muscodor albus
  2. 2. Avenue of the giants
  3. 3. Honduran Rainforest With High Plant Diversity Comes High Microbial Diversity
  4. 4. Cinnamomum zeylanicum
  5. 5. Phytophageous Mites
  6. 6. Anti-Mite Measures
  7. 7. SEM of the fungus
  8. 8. Maximum Parsimony Phenogram of 18S r DNA Sequences of Xylariales
  9. 9. Muscodor albus Properties No spores 2.Ropy mycelium 3.Strange odor 4.Antibiotic activity US Patent-6911338 5.Related to xylaria
  10. 10. Bioassay of VOC’s of M. albus
  11. 11. Trapping the VOC’s of M. albus Divinylbenzene/carburen on polydimethylsiloxane on a stable flex fibre
  12. 12. Joe Sears with GC/MS
  13. 13. The VOC’s of M. albus
  14. 14. Bioassay of Fungal VOC’s
  15. 15. Bioactivity of M. albus and its VOC‘s
  16. 16. Bioactivity of M. albus VOC’s Test Organism Alcohols Esters Ketones Acids Lipids 0.48 l/CC 0.53 l/CC 0.02 l/CC 0.09 l/CC 0.08l/CC % growth of % growth of % growth of % growth of % growth of control control control control control Pythium ultimum 11.2  4 0 67.5  7 40.9  3 75  0 Rhizoctonia solani 55 5 0 67.57.5 67.57.5 400 Tapesia yallundae 3515 0 75  25 100 0 1000 Xylaria sp. 7525 0 1000 1000 1000 Sclerotinia sclerotiorum 293 8.11.5 20.612 400 782 Cercospora beticola 588 55 1000 8317 1000 Fusarium solani 7010 55 5 9010 8020 8010
  17. 17. Proton Transfer Reaction Mass Spectrometer
  18. 18. The PTR – MS of Standard VOCs of M. albus Compound Major ion and % in ( ) Acetic acid, 2-phenylethyl 43 (37), 61 (63) ester Phenyl ethyl alcohol 105 (100) Propanoic acid, 2-methyl, 41 (7), 43 (18), 57 (35), 89 (31), 145 (9) 2-methylpropyl ester Propanoic acid, 2-methyl, 43 (39), 71 (42), 89 (16), 159 (3) 3-methylbutyl ester Propanoic acid, 2 methyl, 43 (4), 89 (11), 117 (85) ethyl ester Propanoic acid,2-methyl 41 (11), 43 (42), 89 (47) Propanoic acid, 2-methyl, 43 (3), 103(97) methyl ester 1-Butanol, 3-methyl, 41 (10), 43 (47), 71 (37), 131 (4) acetate
  19. 19. 34 Temperature 800 33 mass 131 32 Temperature (C) 600 Intensity (cps) 31 400 30 mass 103 29 200 Mass 131= Ca. 18ppb 28 Mass 103=Ca. 12ppb 0 14 15 16 17 18 19 Time (days)
  20. 20. Treating Barley Seeds with M. albus
  21. 21. Control of Loose Smut by M. albus Ustilago hordei on barley
  22. 22. Protective effects of M. albus
  23. 23. Preserving Garbage with M. albus VOCs
  24. 24. Tons of Muscodor albus
  25. 25. Muscodor albus in action- decontaminating human waste products
  26. 26. Phillips Toilet on the North Col of Mt Everest
  27. 27. Access to the Upper Amazon of Peru
  28. 28. M. vitigenes from Paullinia paullinioides
  29. 29. Muscodor roseus from Ironwood in Australia
  30. 30. S.E Asia Tesso Nilo collecting site
  31. 31. Collecting in the Tesso Nilo Area of Sumatra, Indonesia
  32. 32. M. albus Isolate I-41 Sumatra, Indonesia
  33. 33. Coastal Ecuador- Dry Forest
  34. 34. Fun in the jungle
  35. 35. A Jungle Nasty-fer de Lance Percy’s foot
  36. 36. M. albus from Guazuma ulmifolia in the dry forest Of coastal Ecuador
  37. 37. Table 1. GC/MS analysis of the volatile compounds produced by M. albus E-6. RT Total M/z Possible compound MW Area (%) 4:58 32.4 102 *Propanoic acid, 2- 102 methyl-methyl ester 7:07 1.2 116 Butanoic acid, 2-methyl- 116 7:24 1.0 116 Butanoic acid, 3-methyl- 116 9:35 1.3 84 2-Butenal, 2-methyl- 84 10:19 28.0 130 *1 Butanol, 2-methyl- 130 12:20 5.9 158 Butanoic acid, 3- 158 methylbutyl ester 12:24 4:2 158 *Propanoic acid, 2- 158 methyl, 2 -methylbutyl ester 13:51 1.1 118 Propanoic acid, 2- 118 hydroxy-2-methyl-methyl ester 14:07 1.0 86 3-Buten-1-ol, 3-methyl- 86 15:36 1.6 140 1-Octene, 3-ethyl- 140 16:12 1.1 142 *4-Nonanone 142 18:21 1.4 204 Naphthalene,decahydro- 204 4a-methyl-1-methylene-7- (1methylethylidine)-,(4aR- trans) 19:54 1.0 204 1H- 204 cycloprop[e]azulene,1a,2, 3,5,6,7,7a,7b-octahydro- 1,1,4,7-tetramethyl-,[1aR- (1a.alpha.)] 20:02 3.8 222 Guaiol 222 20:25 5.7 204 *Caryophyllene 204 22:33 100 88 *Propanoic acid, 2- 88 methyl- 24:36 1.8 101 Formamide,N-(1- 101 methylpropyl) 24:55 1.0 98 2-Furanmethanol 98 25:57 1.0 204 *Azulene, 1,2,3,5,6,7,8, 204
  38. 38. Muscodor crispans. from Ananas ananasoides (Bolivan Amazon)
  39. 39. Retention Time Min. Total Area Possible Compound M- Da 2:05 0.139 Acetaldehyde 44.03 VOCs 3:40 3:51 0.623 0.283 Ethyl Acetate 2-Butanone 88.05 72.06 B-23 4:08 3.056 Propanoic acid, 2-methyl-, methyl ester 102.07 4:18 1.241 Ethanol 46.04 5:29 0.229 Acetic acid, 2-methylpropyl 116.08 ester 6:39 0.109 Propanoic acid, 2-methyl-, 2- 144.12 methylpropyl ester 6:46 0.178 1-Propanol, 2-methyl- 74.07 6:52 0.151 2-Butenal, 2-methyl-, (E)- 84.06 7:12 0.479 1-Butanol, 3-methyl-, acetate 130.10 8:18 0.301 Hexane, 2,3-dimethyl- 114.14 8:21 0.478 Propanoic acid, 2-methyl-, 2- 158.13 methylbutyl ester 8:31 1.538 1-Butanol, 3-methyl- 88.09 13:37 35.118 Propanoic acid, 2-methyl- 88.05 14:41 0.394 Formamide, N-(1- 101.08 methylpropyl)- 16:44 0.131 Acetic acid, 2-phenylethyl 164.08 ester 20:44 0.720 Cyclohexane, 1,2-dimethyl- 192.19 3,5-bis(1-methylethenyl)-
  40. 40. Effects of B-23 on human pathogenic bacteria Organism Type of Exposure Growth/ No Comments Cell Wall Time Growth (in the presence of M. crispans) S. aureus 6538 Gram + 2, 4 and 6 No growth days S. cholerasuis 10708 Gram - 2, 4 and 6 No growth days P. aeruginosa 15442 Gram - 2 days Growth No visible difference between exposed and control plates. S. aureus ATCC 43300 Gram + 2, 4 and 6 Growth No actual colonies formed, (MRSA) days just a slightly filmy growth. Y. pestis 91-3365 Gram - 3 and 5 days No growth B. anthracis A2084 Gram + 3 and 5 days Growth Only a few colonies left after exposure and when incubated, more grew. M. tuberculosis 3081 Acid-fast 2, 4, 7 and 14 No growth (resistant to isoniazid) days M. tuberculosis 50001106 Acid-fast 2, 4, 7 and 14 No growth (resistant to streptomycin) days M. tuberculosis 59501228 Acid-fast 2, 4, 7 and 14 No growth (resistant to days streptomycin/ethambutol) M. tuberculosis 59501867 Acid-fast 2, 4, 7 and 14 No growth (susceptible) days
  41. 41. Distribution of Muscodor spp. in the World
  42. 42. Plant (Family)Sources of Muscodor albus 1. Lauraceae 2. Myristicaceae 3. Proteaceae 4. Combretaceae 5. Sapindaceae 6. Leguminosae 7. Caesalpiniaceae 8. Bromeliaceae
  43. 43. Lessons Learned from M. albus and its relatives 1. Each has 95% -99% 18S rDNA partial sequence similarity to the original isolate of M. albus -620. 2. They make different VOC’s and in differing amounts. 3. They are found as endophytes in the wet tropical regions of the world from Thailand, to Indonesia, Australia, Venezuela, and Peru. They are confined to +/- 16 degrees from the equator. 4, Each has some type of bioactivity. 5. Some are being developed for commercial purposes. 6. Many other surprises remain in the jungle.
  44. 44. Northern Patagonia –Land of the Alerces
  45. 45. Torres del Paine
  46. 46. Calving of a glacier in a hanging glacier-Chile
  47. 47. Ulmo and Northern Patagonia
  48. 48. An Endophytic Gliocladium sp. from Eucryphia cordifolia in Northern Patagonia Isolated by the M. albus selection technique
  49. 49. SEM’s of Gliocladium sp.
  50. 50. The Hydrocarbons of Gliocladium sp. Relati Molecu Grown on Oat meal agar 18 days under minimal oxygen ve Possible lar Time Area Compound Weight 1.603 1.213 Oxirane, ethyl- 72.06 7.648 1.807 1-Butanol, 3-methyl-, acetate 130.10 2.081 1.419 Heptane, 2-methyl- 114.14 7.836 1.928 2-n-Butyl furan 124.09 2.666 2.519 Octane 114.14 8.026 0.279 Benzene, 1 3-dimethyl- 106.08 3.138 0.261 1-Octene 112.13 8.114 0.368 Decane, 3, 3, 5-trimethyl- 184.22 4.598 7.132 Ethanol 46.04 8.303 0.335 Pentane, 1-iodo- 197.99 4.872 1.133 Cyclohexene, 4-methyl- 96.09 8.364 1.379 2-Hexanol 102.10 5.204 0.342 Hexane, 2, 4-dimethyl- 114.14 8.498 0.306 Acetic acid, pentyl ester 130.10 5.378 0.180 Undecane, 2, 6-dimethyl- 184.22 8.735 1.228 Hexanoic acid, methyl ester 130.10 5.533 0.504 Hexadecane 226.27 9.066 7.956 1-Butanol, 3-methyl- 88.09 Heptane, 5-ethyl-2, 2, 3- 9.302 0.134 Phenol, 4-ethyl- 122.07 5.941 0.564 trimethyl- 170.20 9.817 0.710 3-Octanone 128.12 6.365 0.476 Undecane, 4-methyl- 170.20 10.054 1.780 Acetic acid, hexyl ester 144.12 Heptane, 5-ethyl-2, 2, 3- 6.418 0.180 trimethyl- 170.20 10.708 0.143 2-Heptanol 116.12 Octane, 3-ethyl-2, 7- 10.985 0.574 7-Octen-2-one 126.10 6.668 0.155 dimethyl- 170.20 11.242 6.514 Cyclopropane, propyl- 84.09 6.768 1.021 Decane, 2, 2, 6-trimethyl- 184.22 11.329 0.550 Acetic acid, sec-octyl ester 172.15 6.931 0.360 Undecane 156.19 11.545 11.294 Acetic acid, heptyl ester 158.13 7.112 0.195 Decane, 3, 3, 5-trimethyl- 184.22 11.775 0.205 Octanoic acid, methyl ester 158.13 7.173 0.592 Nonane, 3-methyl- 142.17 11.938 0.485 3, 5-Octadiene (Z, Z) 110.11 7.232 0.601 1-Propanol, 2-methyl- 74.07 12.265 3.289 1-Butanol, 3-methyl-, acetate 131.11 7.325 0.746 Furan, 4-methyl-2-propyl- 124.09
  51. 51. Hydrocarbons ..continued 12.878 11.533 Acetic acid 60.02 12.931 12.008 Acetic acid, octyl ester 172.15 13.381 0.222 3-Octen-2-ol (Z) 128.12 13.584 0.176 Neoisolongifolene 204.19 14.455 0.130 Propanic acid, 2-methyl- 88.05 14.543 0.134 Cycloheptanone, 2-methylene- 124.09 14.651 0.651 1H-Indene, octahydro-, cis 124.13 Cyclopentadiene, 5, 5-dimethyl-1- 14.926 0.254 ethyl- 122.11 15.335 0.137 Butyrolactone 86.04 15.477 0.147 Acetic acid, decyl ester 200.18 15.673 0.465 Pentanoic acid, 3-methyl- 116.08 15.835 0.271 Cyclodecene 138.14 16.472 0.174 Pentanoic acid 102.07 17.653 1.657 Hexanoic acid 116.08 18.360 1.073 Phenylethyl alcohol 122.07 19.588 0.355 Phenol, 4-ethyl-2methoxy- 152.08
  52. 52. Gliocladium a virtual chemistry factory
  53. 53. Victoria, Australia temperate rainforest
  54. 54. The stages of Ascocoryne
  55. 55. The Victoria Fires of 09
  56. 56. Could fungi have contributed to the formation of crude oil ?? SEM’s of Gliocladium sp. and a fossilized fungus
  57. 57. Chief Contributors

×