Study of the genetic diversity of the genus Passiflora L. and its distribution in Colombia<br />John Ocampo<br />École Sup...
People involved in the study<br />CIRAD<br /><ul><li>  Philippe FELDMANN
  Geo COPPENS d’EECKENBRUGGE
Ange-Marie RISTERUCCI</li></ul>BIOVERSITY INTERNATIONAL – CIAT<br /><ul><li>Andrew JARVIS
  Xavier SCHELDEMAN</li></ul>Ginés-Mera Fellowship<br />
Context<br /><ul><li>Genetic resources in Colombia (cultivated species)
Passion fruit priorities:</li></ul>	- Collection and characterization <br />	- Taxonomy problems<br />	- Conservation prob...
General objective<br />To study the genetic diversity of the genus Passiflora L. and its biogeographic distribution in Col...
Specific objectives<br /><ul><li>To map the distribution and the diversity of Colombian Passifloraceae species using Geogr...
To characterize the morphologic and molecular diversity at inter- and intra-specific levels.
To evaluate the potential of Passiflora as an indicator group to evaluate the risks of biodiversity erosion and take them ...
1. Passifloraceae family Taxonomy<br /> 18 genera<br />ca. 650 species<br />America <br />   Africa<br />Asia<br />  Killi...
Subgenus ASTROPHEA (Feuillet & MacDougal, 2003)<br /><ul><li> Astrophea</li></ul>- Arborescent<br />- American<br />- 2n =...
SubgenusDECALOBA (Feuillet & MacDougal, 2003)<br /><ul><li>Decaloba
Psilanthus
Murucuja
Pseudomurucuja
Apodogyne</li></ul>- Climbers<br />- America<br />  South Asia and Australia <br />- 2n = 12, 24, 36, 72<br />Killip (1938...
SubgenusPASSIFLORA (Feuillet & MacDougal, 2003)<br /><ul><li>Passiflora
Tacsonia
Distephana
Manicata
Dysosmia</li></ul>- Climbers<br />Killip (1938)<br />- American<br />- 2n = 18, 20 (Dysosmia)<br />
SubgenusDEIDAMIOIDES (Feuillet & MacDougal)<br /><ul><li>Deidamioides
Tryphostemmatoides</li></ul>- Climbers<br />- American<br />- 2n = ??<br />Killip<br />
2. Biology of reproduction<br /><ul><li>Allogamous: cross-pollination
Cytogenetic:2n = 12, 18, 20, 24, 36, 72</li></ul>                                 x = 6 y 9<br /><ul><li>Heredability:pate...
Pollinationsyndromes<br />
3. Distribution of thegenusPassifloraL.<br />22 species<br />Subg. Decaloba<br />
4. Uses<br />Other…<br />Ornamental<br />Food<br />Pharmacopeia<br />Liquors<br />Medicinal<br />
5. Cultivatedspecies in Colombia<br />
Economic importance – International markets<br />P. edulis f. flavicarpa<br />Yellow maracuja - Maracuyá<br />P.  ligulari...
Economic importance – International markets<br />P. tripartita var. mollissima<br />Curuba de Castilla<br />P. tarminiana<...
Economic importance – International markets<br />P. quadrangularis<br />Giant granadilla - Badea<br />P. popenovii<br />Gr...
Study Components<br />I- Diversity distribution in Colombia (GIS)<br />	1.a. Biogeography and conservation<br />	1.b. Pote...
I – Diversity Distribution in Colombia (SIG)<br />1a. Biogeography and conservation of Colombian Passifloraceae <br />.<br />
Reported species <br />HERBARIA<br />National<br />PSO  - HAU - COL  - FAUC - CUVC - FMB <br />VALLE - MEDEL - CHOCO – TOL...
Maximum Diversity in the American Andean Region, especially in Colombia and Ecuador<br />Diversity Centre<br />Diversity d...
Species spatial distribution<br />High density in the central mountain region.<br />Low density North-East and South of th...
Expeditions and collected samples<br /><ul><li> Species area richness and collection gaps
 Access permits (MMA - Ministerio del Medio Ambiente)
 Public order</li></ul>555 collections from 17 of 32 departments<br />TOTAL : 3.930 OBSERVATIONS<br />
Elevation<br />Species distribution by Biogoegraphic region<br />Caribbean      (38)<br />Diversity concentrated in the An...
Pacific<br />Andes<br />Caribbean<br />Amazonia<br />Orinoquia<br />Colombian Biogeography & Passifloracea Species <br />
Passifloraceae distribution in Colombia <br />(Maximum Distance vs. Circular Area)<br />Species of continental distributio...
f. Threat status of Passifloraceae<br /><ul><li> Circular area CA50
 Maximum Distance MaxD
 Number of records
 Date of records</li></ul>UICN Categories<br />EX:Extinct		<br />EW:Extinct in the wild<br />CR:Critically endangered<br /...
1. Diversity and potentiel distribution<br />1b. Diversity distribution and in situ conservation of the Colombians Passifl...
Software DIVA-GIS<br />Speciesrichness and diversity<br />Herbarium data: 3.330<br />Literature data: 45<br />Collection d...
1<br />2<br />Nine ultra- diverse zones in the Andes, four sub-explored (1, 6, 8, 9)<br />3<br />4<br />Species richness<b...
Protectedareas<br />In situ conservation<br />“Hotspot”<br />
Ultra-diverse zones vs. Coffee growing ecotopes<br />Small reserves and micro-basin management<br />Conservation strategie...
Crop systems</li></ul>Software DIVA-GIS<br />
Conclusions<br /><ul><li>The highest number of species are located between 1.000 and 2.000 m in fragmentation forests.
Nine ultra-diverse (hotspot) and non-protected zones in the Andes.
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Study of the genetic diversity of the genus Passiflora L. and its distribution in Colombia

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Workshop Ginés-Mera fellowship , Cali - Colombia (may 12, 13 and 14)
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Study of the genetic diversity of the genus Passiflora L. and its distribution in Colombia

  1. 1. Study of the genetic diversity of the genus Passiflora L. and its distribution in Colombia<br />John Ocampo<br />École Supérieure d’Agronomie de Montpellier<br />Université de Montpellier II<br />Ph.D.<br />ocampo.john@gmail.com<br />
  2. 2. People involved in the study<br />CIRAD<br /><ul><li> Philippe FELDMANN
  3. 3. Geo COPPENS d’EECKENBRUGGE
  4. 4. Ange-Marie RISTERUCCI</li></ul>BIOVERSITY INTERNATIONAL – CIAT<br /><ul><li>Andrew JARVIS
  5. 5. Xavier SCHELDEMAN</li></ul>Ginés-Mera Fellowship<br />
  6. 6. Context<br /><ul><li>Genetic resources in Colombia (cultivated species)
  7. 7. Passion fruit priorities:</li></ul> - Collection and characterization <br /> - Taxonomy problems<br /> - Conservation problems<br />
  8. 8. General objective<br />To study the genetic diversity of the genus Passiflora L. and its biogeographic distribution in Colombia.<br />
  9. 9. Specific objectives<br /><ul><li>To map the distribution and the diversity of Colombian Passifloraceae species using Geographic Information Systems (GIS).
  10. 10. To characterize the morphologic and molecular diversity at inter- and intra-specific levels.
  11. 11. To evaluate the potential of Passiflora as an indicator group to evaluate the risks of biodiversity erosion and take them into account in the development of strategies for in situ genetic resources.</li></li></ul><li>Introduction<br />1. Taxonomy<br />2. Reproduction Biology<br />3. Distribution<br />4. Uses<br />5. Cultivated species<br />
  12. 12. 1. Passifloraceae family Taxonomy<br /> 18 genera<br />ca. 650 species<br />America <br /> Africa<br />Asia<br /> Killip (1938)<br />22 subgenera<br />GenusPassifloraL.<br />ca. 573 species<br />America<br /> Asia<br />Feuillet & McDougal (2003)<br />4 subgenera<br />Astrophea DecalobaDeidamioides Passiflora<br />
  13. 13. Subgenus ASTROPHEA (Feuillet & MacDougal, 2003)<br /><ul><li> Astrophea</li></ul>- Arborescent<br />- American<br />- 2n = 24<br />Killip (1938)<br />
  14. 14. SubgenusDECALOBA (Feuillet & MacDougal, 2003)<br /><ul><li>Decaloba
  15. 15. Psilanthus
  16. 16. Murucuja
  17. 17. Pseudomurucuja
  18. 18. Apodogyne</li></ul>- Climbers<br />- America<br /> South Asia and Australia <br />- 2n = 12, 24, 36, 72<br />Killip (1938)<br />
  19. 19. SubgenusPASSIFLORA (Feuillet & MacDougal, 2003)<br /><ul><li>Passiflora
  20. 20. Tacsonia
  21. 21. Distephana
  22. 22. Manicata
  23. 23. Dysosmia</li></ul>- Climbers<br />Killip (1938)<br />- American<br />- 2n = 18, 20 (Dysosmia)<br />
  24. 24. SubgenusDEIDAMIOIDES (Feuillet & MacDougal)<br /><ul><li>Deidamioides
  25. 25. Tryphostemmatoides</li></ul>- Climbers<br />- American<br />- 2n = ??<br />Killip<br />
  26. 26. 2. Biology of reproduction<br /><ul><li>Allogamous: cross-pollination
  27. 27. Cytogenetic:2n = 12, 18, 20, 24, 36, 72</li></ul> x = 6 y 9<br /><ul><li>Heredability:paternal / biparental</li></ul>Muschner et al. (2006) Hansen et al. (2007)<br /><ul><li>Heteroplasmy</li></ul>Mráček (2005) <br />
  28. 28. Pollinationsyndromes<br />
  29. 29. 3. Distribution of thegenusPassifloraL.<br />22 species<br />Subg. Decaloba<br />
  30. 30. 4. Uses<br />Other…<br />Ornamental<br />Food<br />Pharmacopeia<br />Liquors<br />Medicinal<br />
  31. 31. 5. Cultivatedspecies in Colombia<br />
  32. 32. Economic importance – International markets<br />P. edulis f. flavicarpa<br />Yellow maracuja - Maracuyá<br />P. ligularis<br />Sweet granadilla<br />P. edulis f. edulis<br />Purple maracuja - Gulupa<br />
  33. 33. Economic importance – International markets<br />P. tripartita var. mollissima<br />Curuba de Castilla<br />P. tarminiana<br />Curuba India<br />P. maliformis<br />Cholupa<br />
  34. 34. Economic importance – International markets<br />P. quadrangularis<br />Giant granadilla - Badea<br />P. popenovii<br />Granadilla de Quijos<br />P. alata<br />Maracuja doce<br />
  35. 35. Study Components<br />I- Diversity distribution in Colombia (GIS)<br /> 1.a. Biogeography and conservation<br /> 1.b. Potential distribution and adaptation<br />II- Morphological diversity (descriptors)<br />III- Genetic diversity (CAPS – DNAcp/mt)<br />
  36. 36. I – Diversity Distribution in Colombia (SIG)<br />1a. Biogeography and conservation of Colombian Passifloraceae <br />.<br />
  37. 37. Reported species <br />HERBARIA<br />National<br />PSO - HAU - COL - FAUC - CUVC - FMB <br />VALLE - MEDEL - CHOCO – TOLI - CAUP <br />AFP - UIS – CDMB - COAH – JAUM - HUQ <br />SURCO<br />Foreign<br />P : Museum d’Histoire Naturelle – Paris – France<br />MO : Missouri Botanical Garden – USA<br />MA : Real Jardin Botánico – Madrid – Espagne<br />K :Kew Garden – London - Grande-Bretagne<br />NY : New York Botanical Garden– USA<br />LITERATURE<br />Killip (1938, 1960), Escobar (1988, 1989) etc.<br />3.375 georeferenced data<br />
  38. 38. Maximum Diversity in the American Andean Region, especially in Colombia and Ecuador<br />Diversity Centre<br />Diversity distribution of the American Passion Fruits<br />3.375 georeferenced data<br />16<br />4<br /> 23.5º N<br />167 reported species (26 new species for the country)<br /> Tropic of Cancer<br />20<br />14<br />20<br />70<br />15<br />17<br />10<br />10<br />18<br />55<br />25<br />13<br />78<br />31<br />48<br />53<br />30<br />32<br />36<br />167<br />0º<br />90<br />127<br />73<br />48<br />Tropic of Capricorn<br />2<br />16<br /> -23.5º S<br />19<br />3<br />
  39. 39. Species spatial distribution<br />High density in the central mountain region.<br />Low density North-East and South of the Andes.<br />Very marked low density in lowlands, especially in the Orinoquian and Amanzonian regions.<br />DIVA-GIS software<br />
  40. 40. Expeditions and collected samples<br /><ul><li> Species area richness and collection gaps
  41. 41. Access permits (MMA - Ministerio del Medio Ambiente)
  42. 42. Public order</li></ul>555 collections from 17 of 32 departments<br />TOTAL : 3.930 OBSERVATIONS<br />
  43. 43. Elevation<br />Species distribution by Biogoegraphic region<br />Caribbean (38)<br />Diversity concentrated in the Andean Region<br />Pacific (36)<br />Andes (123)<br />Orinoquia (19)<br />Amazonia (44)<br />
  44. 44. Pacific<br />Andes<br />Caribbean<br />Amazonia<br />Orinoquia<br />Colombian Biogeography & Passifloracea Species <br />
  45. 45. Passifloraceae distribution in Colombia <br />(Maximum Distance vs. Circular Area)<br />Species of continental distribution<br />Species of regional distribution and common endemic species<br />R2= 0.779<br />P. vitifolia<br />P. foetida<br />P. auriculata<br />P. quadrangularis<br />P. nitida<br />P. maliformis<br />P. mollis<br />P. mixta<br />Endemic species <br />P. cumbalensis<br />P. menispermifolia<br />P. ligularis<br />P. suberosa<br />P.arborea<br />P. oerstedii<br />P. capsularis<br />P. seemannii<br />P. coriaceae<br />P.misera<br />P. biflora<br />P. spinosa<br />P..coccinea<br />P. rubra<br />P. antioquiensis<br />P. micropetala<br />P. serratodigitata<br />P. involucrata<br />D. parviflora<br />1000<br />400<br />
  46. 46. f. Threat status of Passifloraceae<br /><ul><li> Circular area CA50
  47. 47. Maximum Distance MaxD
  48. 48. Number of records
  49. 49. Date of records</li></ul>UICN Categories<br />EX:Extinct <br />EW:Extinct in the wild<br />CR:Critically endangered<br />EN:Endangered <br />VU:Vulnerable<br />NT:Near threatened<br />LC:Least concern<br />DD:Deficient Data<br />
  50. 50. 1. Diversity and potentiel distribution<br />1b. Diversity distribution and in situ conservation of the Colombians Passifloraceae.<br />
  51. 51. Software DIVA-GIS<br />Speciesrichness and diversity<br />Herbarium data: 3.330<br />Literature data: 45<br />Collection data: 555<br />3.930 georeferenced data<br />
  52. 52. 1<br />2<br />Nine ultra- diverse zones in the Andes, four sub-explored (1, 6, 8, 9)<br />3<br />4<br />Species richness<br />5<br />6<br />7<br />8<br />9<br />Species potential distribution<br />“Hotspot”<br />Software DIVA-GIS<br />
  53. 53. Protectedareas<br />In situ conservation<br />“Hotspot”<br />
  54. 54. Ultra-diverse zones vs. Coffee growing ecotopes<br />Small reserves and micro-basin management<br />Conservation strategies<br /><ul><li>Corridor creation
  55. 55. Crop systems</li></ul>Software DIVA-GIS<br />
  56. 56. Conclusions<br /><ul><li>The highest number of species are located between 1.000 and 2.000 m in fragmentation forests.
  57. 57. Nine ultra-diverse (hotspot) and non-protected zones in the Andes.
  58. 58. There is no geographic relation between richness and endemism.
  59. 59. There is a need for urgent conservation strategies.
  60. 60. Of the 167 recorded species, 42 have edible fruits and most are not used. </li></li></ul><li>II. Morphological diversity in the genus Passiflora L. <br />
  61. 61. Collection and characterization (ex situ conservation)<br /><ul><li> Collections
  62. 62. Tenerife (El Cerrito – Valle del Cauca) – 2800 m
  63. 63. El Moral (El Cerrito – Valle del Cauca) – 2400 m
  64. 64. San Joaquín (El Tambo – Cauca) – 1800 m
  65. 65. Paraguacito (Buenavista – Quindío) – 1250 m
  66. 66. Arauca (Risaralda – Caldas) – 1000 m
  67. 67. In situ (collections)
  68. 68. Plant material</li></ul> 124 accessions <br /> 60 species<br /> 9 subgenera (Killip, 1938)<br />
  69. 69. List of 127 descriptors<br />
  70. 70. Relative variance components – Quantitative characters<br />50<br />
  71. 71. d. Factor loadings from a Principal Component Analysis - PCA<br />COMPONENTS<br />Descriptors<br />1<br />2<br />3<br />4<br />5<br />ACP - Factor loadings<br /> 83% total variance<br />-0.088<br />0.215<br />-0.023<br />0.041<br />-0.892<br />STDI<br />0.759<br />LEMS<br />0.353<br />0.365<br />0.152<br />0.041<br />LELC<br />0.019<br />0.571<br />-0.686<br />0.009<br />-0.195<br />-0.017<br />0.040<br />0.011<br />0.014<br />LENN<br />0.962<br />PENN<br />0.648<br />0.407<br />0.013<br />-0.055<br />0.158<br />0.135<br />-0.287<br />0.010<br />0.145<br />PDDI<br />0.847<br />PDLF<br />-0.943<br />-0.054<br />-0.136<br />-0.007<br />-0.044<br />-0.054<br />-0.113<br />-0.006<br />-0.046<br />PDBS<br />-0.973<br />BRLR<br />0.723<br />0.296<br />0.186<br />-0.091<br />0.240<br />FLPL<br />0.774<br />0.501<br />-0.021<br />0.235<br />0.042<br />Cp.1: Flower length<br />0.595<br />-0.024<br />0.095<br />0.028<br />FLPW<br />0.713<br />FLSL<br />0.716<br />0.577<br />0.036<br />0.243<br />-0.028<br />0.520<br />0.034<br />0.074<br />-0.108<br />FLSW<br />0.754<br />Cp.2: Flower width and bract shape<br />FLLE<br />0.960<br />0.179<br />0.045<br />0.017<br />0.090<br />0.063<br />0.086<br />-0.100<br />0.119<br />0.904<br />FLHL<br />FLHL<br />0.824<br />0.208<br />0.026<br />-0.080<br />-0.137<br />FLCN<br />0.723<br />-0.521<br />0.034<br />-0.007<br />-0.329<br />Cp.3: Stem with, peduncle branching and leaf length<br />0.529<br />0.651<br />0.082<br />0.097<br />0.171<br />FLFL<br />FLSF<br />0.677<br />0.658<br />0.076<br />0.061<br />-0.049<br />0.454<br />0.693<br />0.067<br />0.073<br />0.263<br />FLOL<br />0.064<br />0.047<br />0.033<br />0.099<br />0.964<br />FLSL<br />0.236<br />0.099<br />0.363<br />0.214<br />0.711<br />FLOP<br />Cp.4:Number of nectaries on leaf margin<br />-0.160<br />-0.221<br />-0.131<br />0.241<br />BRWI/BRLE<br />0.718<br />0.699<br />FLNC/FLHD<br />0.056<br />0.042<br />0.043<br />0.368<br />Expl.Var<br />7.610<br />6.496<br />3.336<br />1.256<br />1.238<br />Prp.Totl<br />0.317<br />0.271<br />0.139<br />0.052<br />0.052<br />Cp.5:Leaf serration<br />% total varianza<br />46.028<br />16.999<br />11.138<br />4.913<br />3.986<br />
  72. 72. Manicata<br />Distephana<br />PCA – Three-dimensional plot of the scores for Passiflora accessions<br />Tacsonia<br />Passiflora<br />Psilanthus<br />Dysosmia<br />Decaloba<br />Tryphostemmatoides<br />Astrophea<br />
  73. 73. 32 Discriminant qualitative characters between subgenera<br />
  74. 74. e. Qualitative data dendrogram (nNeighbour Joining, 32 characters)<br />n = 12<br />n = 6<br />n = 10<br />n = 9<br />n = 6<br />n = ??<br />Distances Sokal & Michener<br />
  75. 75. Conclusions<br />The new descriptors list has permitted to rank species together with taxonomy in a coherent way. However, there are some exceptions.<br />The analysis is coherent with Killip's classification (1938) at the subgenus level.<br />Descriptors related to the flower permit a higher discrimination between subgenera and species.<br />
  76. 76. III. Genetic diversity (CAPS – DNAcp/mt)<br />
  77. 77. Taxon sampling<br /><ul><li> 213 individuals
  78. 78. 151 species
  79. 79. 15 subgenera (Killip, 1938)</li></ul>Origin<br /><ul><li> Collections in Colombia – 75 %
  80. 80. Passion Fruit National Collections – Blois (France) – 22 %
  81. 81. Royal Botanical Garden – Meise (Belgium) – 2 %
  82. 82. Cameroon and Gabon - 1% (outgroup)</li></li></ul><li>Tecnic – Markers RFLP-PCR (2 couple of primers / 6 enzymes)<br />
  83. 83. Outgroup<br />Previousstudies<br />PASSIFLORA<br />Yockteng & Nadot (2004)<br />Nuclear Sequences<br />ncpGS<br /> - 91 species<br /> - 17 subgenera (Killip, 1938)<br />DECALOBA<br />ASTROPHEA<br />
  84. 84. Previousstudies<br />PASSIFLORA<br />Hansen et al. (2006)<br />Chloroplas Sequences<br />TrnL / TrnT<br />- 57 species<br />- 16 subgenera (Killip, 1938)<br />DECALOBA<br />outgroup<br />ASTROPHEA<br />
  85. 85. Cytoplasmatic genome inter-specific variation<br />Chloroplast<br />280 haplotypes<br />Mitochondria<br />372 haplotypes<br />Chlorotypes are easier to interpret than mitotypes<br />
  86. 86. Observed variation per primer-enzyme combination (haplotypes)<br />Mitochondrial Chloroplastic<br />
  87. 87. AFTD of Chloroplastic data<br />OUTGROUP<br />
  88. 88. Tryphostemmatoides<br />Astrophea<br />- Three big groups<br />- Genus monophyly ?<br />Chloroplast DNA<br /><ul><li> Decaloba – much better separated and structured
  89. 89. Passiflora - little structure
  90. 90. Astrophea
  91. 91. Tryphostemmatoides</li></ul>NJ, distance de Sokal & Michener<br />
  92. 92. Tacsonia<br />Astrophea<br /><ul><li> Strong approximation of Astrophea and Tryphostemmatoides to Passiflora</li></ul>Tryphostemmatoides<br /><ul><li> Little cohesion with morphology at the section and series levels.</li></ul>Tacsonia<br /><ul><li> Geographic structuring by:</li></ul> - altitude range (Tacsonia, stip. ren.)<br /> - region (Tacsonia, Ecuador/Colombia).<br />Mitochondrial DNA<br /><ul><li>Separation of the Decaloba group, better structured
  93. 93. Genus monophyly ?</li></li></ul><li>Conclusions<br /><ul><li> Divergence between morphologic, chloroplastic and mitochondrial data explains plastid transmission modes:</li></ul>Chloroplastic genome transmission is basically paternal or biparental in the Passiflora group, and maternal in the mitochondiral genome.<br /><ul><li>The doubt on whether the Passifloragenus is not monophyletic imposes a re-examination of the generic classification of Passifloraceae.
  94. 94. The data set must be interpreted taking into account the geological context of tropical flora separation and the formation of the Andean mountain range.
  95. 95. RETICULATE EVOLUTION</li></li></ul><li>Future prospects <br />Passion fruits and biodiversityComplete and verify our knowledge on little or non-exploited zones and tune further geographic analyses to test passion fruits as an indicator group. <br />Passion fruits and genetic resourcesExplore the possibility of interspecific hybridization to compensate for the deficiencies of the main cultivated species.<br />Passifloraceae studies and their evolution- Sequences of other genome regions.- Study hybridization and introgression between sympatric species (compatibility, molecular markers and cytogenetics) to understand cross-linking mechanisms. - Study the plastid genome transmission in the family at the genus level.<br />
  96. 96. Thanks!<br />
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