Study of the genetic diversity of the genus Passiflora L. and its distribution in

Colombia

John OcampoÉcole Supérieure d’Agronomie de Montpellier

Université de Montpellier IIPh.D.

ocampo.john@gmail.com

CIRAD• Philippe FELDMANN

• Geo COPPENS d’EECKENBRUGGE

• Ange-Marie RISTERUCCI

BIOVERSITY INTERNATIONAL – CIAT

• Andrew JARVIS

• Xavier SCHELDEMAN

People involved in the study

Ginés-Mera Fellowship

Genetic resources in Colombia (cultivated species)

Passion fruit priorities:

- Collection and characterization

- Taxonomy problems

- Conservation problems

Context

General objective

To study the genetic diversity of the genus Passiflora L. and its biogeographic distribution in Colombia.

Specific objectives

• To map the distribution and the diversity of Colombian Passifloraceae species using Geographic Information Systems (GIS).

• 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 into account in the development of strategies for in situ genetic resources.

1. Taxonomy

2. Reproduction Biology

3. Distribution

4. Uses

5. Cultivated species

Introduction

1. Passifloraceae family Taxonomy

18 genera

ca. 650 species America

Africa

Asia

Killip (1938)

22 subgeneraGenus Passiflora L.

ca. 573 speciesAmerica

Asia

Feuillet & McDougal (2003)

4 subgenera

Astrophea Decaloba Deidamioides

Passiflora

Subgenus ASTROPHEA (Feuillet & MacDougal, 2003)

Astrophea

- Arborescent

- American

- 2n = 24

Killip (1938)

Subgenus DECALOBA (Feuillet & MacDougal, 2003)

Decaloba Psilanthus Murucuja Pseudomurucuja Apodogyne

- Climbers

- America South Asia and Australia

- 2n = 12, 24, 36, 72

Killip (1938)

Subgenus PASSIFLORA (Feuillet & MacDougal, 2003)

- American

- 2n = 18, 20 (Dysosmia)

Passiflora Tacsonia Distephana Manicata Dysosmia

- Climbers

Killip (1938)

Subgenus DEIDAMIOIDES (Feuillet & MacDougal)

Deidamioides Tryphostemmatoides

- Climbers

- American

- 2n = ??

Killip

2. Biology of reproduction

• Allogamous: cross-pollination

• Cytogenetic : 2n = 12, 18, 20, 24, 36, 72 x = 6 y 9

• Heteroplasmy

Mráček (2005)

Muschner et al. (2006) Hansen et al. (2007)

• Heredability: paternal / biparental

Pollination syndromes

3. Distribution of the genus Passiflora L.

22 species

Subg. Decaloba

4. Uses

Food

Pharmacopeia

Liquors

Medicinal

Ornamental

Other…

5. Cultivated species in Colombia

Economic importance – International markets

P. edulis f. flavicarpa

Yellow maracuja - Maracuyá

P. ligularis

Sweet granadilla

P. edulis f. edulis

Purple maracuja - Gulupa

P. tripartita var. mollissima

Curuba de Castilla

P. tarminiana

Curuba India

P. maliformis

Cholupa

Economic importance – International markets

P. quadrangularis

Giant granadilla - Badea

P. popenovii

Granadilla de Quijos

P. alata

Maracuja doce

Economic importance – International markets

II- Morphological diversity (descriptors)

Study Components

I- Diversity distribution in Colombia (GIS)

1.a. Biogeography and conservation

1.b. Potential distribution and adaptation

III- Genetic diversity (CAPS – DNAcp/mt)

I – Diversity Distribution in Colombia (SIG)

1a. Biogeography and conservation of Colombian Passifloraceae .

Foreign

P : Museum d’Histoire Naturelle – Paris – France

MO : Missouri Botanical Garden – USA

MA : Real Jardin Botánico – Madrid – Espagne

K : Kew Garden – London - Grande-Bretagne

NY : New York Botanical Garden– USA

LITERATURE

Killip (1938, 1960), Escobar (1988, 1989) etc.

HERBARIA

National

PSO - HAU - COL - FAUC - CUVC - FMB

VALLE - MEDEL - CHOCO – TOLI - CAUP

AFP - UIS – CDMB - COAH – JAUM - HUQ

SURCO

Reported species

3.375 georeferenced data

167

127

19

53

90

73

2

48

3

16

3130 3236

48

2555

70 20

10

20 14

10

16

0º

1518

13

17

4

Tropic of Capricorn

Tropic of Cancer

-23.5º S

23.5º N

78

167 reported species (26 new species for the country)

Diversity distribution of the American Passion Fruits

Maximum Diversity in the American Andean Region, especially in Colombia and Ecuador

Diversity Centre

3.375 georeferenced data

DIVA-GIS software

Species spatial distribution

High density in the central mountain region.

Low density North-East and South of the Andes.

Very marked low density in lowlands, especially in the Orinoquian and Amanzonian regions.

Expeditions and collected samples

• Species area richness and collection gaps

• Access permits (MMA - Ministerio del Medio Ambiente)

• Public order

555 collections from 17 of 32 departments

TOTAL : 3.930 OBSERVATIONS

Orinoquia (19)

Amazonia (44)

Caribbean (38)

Andes (123)

Species distribution by Biogoegraphic region

Pacific (36)

Elevation

Diversity concentrated in the Andean Region

Colombian Biogeography & Passifloracea Species

Amazonia

Orinoquia

Caribbean

Pacific

Andes

MaxD (km)

CA

50

(km

2 )

0 200 400 600 800 1000 1200 1400 1600 1800 2000

0

10

20

30

40

50

60

70y = 1.8046+0.0024*x+9.3697E-6*x 2

R2 = 0.7709

P. vitifolia

P. foetida

P. auriculata

P. quadrangularis

P. nitida

P. mollis

P. cumbalensis

P. mixta

P. maliformis

P. serratodigitata

P. antioquiensis

P. oerstedii

P. coriaceae

P. suberosa

P. spinosa

P. involucrata

D. parviflora

P. ligularis

P.arborea

P.misera

P..coccinea

P. micropetala

P. seemannii

P. biflora

P. rubra

P. capsularis

P. menispermifolia

Passifloraceae distribution in Colombia (Maximum Distance vs. Circular Area)

Endemic species

400 1000

MaxD (km)

CA

50

(k

m

2

)

0 200 400 600 800 1000 1200 1400 1600 1800 2000

0

10

20

30

40

50

60

70y = 1.8046+0.0024*x+9.3697E-6*x 2

R2 = 0.7709

R2= 0.779Species of continental distribution

Species of regional distribution and common endemic species

• Circular area CA50

• Maximum Distance MaxD• Number of records• Date of records

UICN Categories

EX: ExtinctEW: Extinct in the wildCR: Critically endangeredEN: EndangeredVU: VulnerableNT: Near threatenedLC: Least concernDD: Deficient Data

f. Threat status of Passifloraceae

1b. Diversity distribution and in situ conservation of the Colombians Passifloraceae.

1. Diversity and potentiel distribution

Species richness and diversity

Literature data: 45

3.930 georeferenced data

Collection data: 555

Herbarium data: 3.330

Software DIVA-GIS

Species richness

Software DIVA-GIS

12

3

4567

9 8

Nine ultra- diverse zones in the Andes, four sub-explored (1, 6, 8, 9)

Species potential distribution

“Hotspot”

Protected areas

In situ conservation

“Hotspot”

Software DIVA-GIS

Ultra-diverse zones vs. Coffee growing ecotopes

Small reserves and micro-basin management

Conservation strategies• Corridor creation • Crop systems

Conclusions

• 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.

• There is no geographic relation between richness and endemism.

• There is a need for urgent conservation strategies.

• Of the 167 recorded species, 42 have edible fruits and most are not used.

II. Morphological diversity in the genus Passiflora L.

Collection and characterization (ex situ conservation)

Collections Tenerife (El Cerrito – Valle del Cauca) – 2800 m El Moral (El Cerrito – Valle del Cauca) – 2400 m San Joaquín (El Tambo – Cauca) – 1800 m Paraguacito (Buenavista – Quindío) – 1250 m Arauca (Risaralda – Caldas) – 1000 m

In situ (collections)

Plant material 124 accessions 60 species 9 subgenera (Killip, 1938)

List of 127 descriptors

ORGAN Qualitative Characters (84) Qunatitative Characters (43)

Stem Form, antocianine, pubescence Internodes

Tendril Antocianine Longitude

Stipule Permanence, antocianine, pubescence and form

Dimensions

Leaf Polymorphism, form, colour, presence, antocianine and pubescence, type, margin

Lobule longitude, angle between lateral rib, petiole—sinus distance, number of margin teeth

Petiole Nectary form Longitude and number of nectaries

Bract Form, union, permanency Dimensions

Flower Form, colours and pubescence of different parts, number and series of filaments, sepal and petal union, antocianine and chlorophyll on sepals and hypanthium

Number, orientation and dimension of different parts

Fruit Pigmentation, pubescence, form, consistency, anthracnosis symptoms

Dimensions, weight

Relative variance components – Quantitative characters

50

ACP - Factor loadingsDescriptors 1 2 3 4 5STDI -0.088 0.215 -0.892 -0.023 0.041LEMS 0.353 0.365 0.152 0.041 0.759LELC 0.019 0.571 -0.686 0.009 -0.195LENN -0.017 0.040 0.011 0.962 0.014PENN 0.648 0.407 0.013 -0.055 0.158PDDI 0.135 0.847 -0.287 0.010 0.145PDLF -0.054 -0.136 -0.943 -0.007 -0.044PDBS -0.054 -0.113 -0.973 -0.006 -0.046BRLR 0.296 0.723 0.186 -0.091 0.240FLPL 0.501 0.774 -0.021 0.235 0.042FLPW 0.713 0.595 -0.024 0.095 0.028FLSL 0.577 0.716 0.036 0.243 -0.028FLSW 0.520 0.754 0.034 0.074 -0.108FLLE 0.960 0.179 0.045 0.017 0.090FLHL 0.904 0.063 0.086 -0.100 0.119FLHL 0.824 0.208 0.026 -0.080 -0.137FLCN -0.521 0.723 0.034 -0.007 -0.329FLFL 0.529 0.651 0.082 0.097 0.171FLSF 0.677 0.658 0.076 0.061 -0.049FLOL 0.454 0.693 0.067 0.073 0.263FLSL 0.964 0.064 0.047 0.033 0.099FLOP 0.711 0.236 0.099 0.363 0.214BRWI/BRLE -0.160 0.718 -0.221 -0.131 0.241FLNC/FLHD 0.699 0.056 0.042 0.043 0.368Expl.Var 7.610 6.496 3.336 1.256 1.238Prp.Totl 0.317 0.271 0.139 0.052 0.052% total varianza 46.028 16.999 11.138 4.913 3.986

COMPONENTS

d. Factor loadings from a Principal Component Analysis - PCA

83% total variance

Cp.2: Flower width and bract shape

Cp.3: Stem with, peduncle branching and leaf length

Cp.4: Number of nectaries on leaf margin

Cp.5: Leaf serration

Cp.1: Flower length

PCA – Three-dimensional plot of the scores for Passiflora accessions

Passiflora

Decaloba

Astrophea

Manicata

DistephanaDysosmia

PsilanthusTacsonia

Tryphostemmatoides

Feuillet & MacDougal (2003) Astrophea Deidamioides

Killip (1938), Escobar (1988), MacDougal (1994) Astrophea Decaloba Psilanthus Passiflora Distephana Dysosmia Tacsonia Manicata TryphostemmatoidesSpecies/accession number n= 3/4 n= 17/31 n= 1/1 n= 19/44 n= 1/2 n= 1/2 n= 16/36 n= 1/1 n= 2/2Main pollinators medium (honey) bees small to large bees - wasps sword-billed large (carpenter) bees hummingbirds bees sword-billed hummingbirds bees

hummingbird hummingbirdChromosome number 2n = 24 2n = 12, 22, 24, 36 2n = 12 2n = 18 2n = 18 2n = 18, 20, 22 2n = 18 2n = 18 2n = 12

Altitudinal range 1.000 - 2.200 m 60 - 2.700 m 2.600 - 3.200 20 - 2.400 m 50 - 1.200 m 30 - 1.200 m 2.100 - 3.700 m 1.900 - 2.500 m 50 - 2.000 m

Descriptors

Habit tree vine vine vine vine vine vine vine vineStem section irregular terete/angular angular terete/angular/winged terete terete terete/angular angular tereteSecondery xylema present absent absent absent absent absent absent absent absentTendril position absent axillary axillary axillary axillary axillary axillary axillary axillary and peduncleStipule short triangular setaceous/linear/foliaceous-aristate setaceous setaceous/linear/foliaceous-aristate linear foliaceous-aristate setaceous/linear/foliaceous-aristate foliaceous-aristate setaceousStipule nectaries (conspicuous) absent absent absent absent/present (P. maliformis ) present present absent absent absentLeaf lobation one three three one/three/more three three one/three three oneLeaf base cuneate-rounded cuneate-rounded/cordate/ cordate cuneate-rounded/cordate cordate cordate cuneate-rounded/cordate cordate rounded

peltate (P. coriacea and P. guatemalensis )Leaf apex obtuse/acute rounded/obtuse/acute acute/very acute rounded to very acute acute acute acute/very acute obtuse to very acute retuseLeaf margin entire entire/serrate (P. adenopoda ) entire entire/serrate serrate serrate serrate serrate entireLaminar nectaries absent present/absent (five species)b present absent absent absent absent absent absentLeaf margin nectaries (conspicuous) absent absent/sinus/ absent absent/sinus sinus absent absent/sinus (P. jardinensis ) absent leaf base

all margin (P. adenopoda ) lateral lobe (P. edulis f. edulis )Petiolar nectaries absent absent/orbicular (P. adenopoda )/ absent all except auriculate orbicular absent linear/orbicular linear absent

cylindrical (P. coriaceae and P. suberosa )/auriculate (P. auriculata )

Dorsal nectaries present absent absent absent absent absent absent absent absentPeduncle branching present absent/present (P. sexflora ) absent absent absent absent absent absent presentBract shape linear setaceous/linear/ linear foliaceous linear pinnatisect foliaceous foliaceous linear

foliaceous (P. adenopoda , P. guatemalensis )/absent (four species)*

Nectary on bract absent absent absent absent/present present absent absent absent absentFlower orientation erect erect/intermediate/pendular pendular erect/intermediate/pendular erect erect erect/intermediate/pendular erect erectCorolla shape reflex intermediate/campanulate campanulate intermediate/reflex reflex intermediate/reflex campanulate/intermediate/reflex reflex reflexDominant corolla color white white red (pink) white/red red white red (including pink or orange) red whiteCorona type filamentous filamentous filamentous filamentous filamentous filamentous tuberculous/ filamentous filamentousCorona filaments sickle-sword-shaped linear linear linear linear linear filamentous, linear linear linear

free free free free fused at base free free free freeNumber of corona series uniseriate biseriate/ uniseriate pentaseriate biseriate 3- to 5-seriate uni- to 5-seriate pentaseriate biseriate

uniseriate (P. guatemalensis and P. adenopoda )/triseriate (P. filipes and P. magadalenae )

Corona color-clear (longest row) white white white red white white/purple purple whiteCorona color-darkest (longest row) yellow white/purple/ white purple red purple purple purple white

yellow (P. guatemalensis ) white (P. guazumaefolia ) white (P. jardinensis )Petals present present/ present present present present present present present

absent (P. coriaceae and P. suberosa )Sepal awn absent absent/present absent present present present present present absentNectar chamber ring present absent absent present present present present present absentHypanthium campanulate flat tubular campalunate tubular campanulate tubular tubular flatLimen present present absent present present present present present presentOvary shape tricostate globose globose globose globose globose globose globose globoseFruit shape globose globose/ elongate globose/ globose globose elongate/ elongate globose

elongate (P. rubra and P. capsularis ) elongate (P. quadrangularis ) globose (P. pinnatistipula )

Categorized (from quantitatives traits)Stem diameter ≥ 120 mm ≤ 20 mm ≤ 20 mm ≥ 30 mm ≥ 30 mm ≤ 20 mm ≥ 30 mm ≥ 30 mm ≤ 20 mm

< 60 mm < 60 mm < 60 mm < 60 mmLeaf length > 215 mm < 190 mm < 190 mm < 190 mm < 190 mm < 190 mm < 190 mm < 190 mm < 190 mm

< 190 mm (P. sphaerocarpa )Sepal length ≤ 30 mm ≤ 30 mm > 30 mm > 30 mm > 30 mm ≤ 30 mm > 30 mm > 30 mm ≤ 30 mm

≤ 30 mm (six species)ª ≤ 30 mm (P. luzmarina and P. mathewsii ) Androgynophore length < 23 mm < 23 mm > 25 mm < 23 mm > 25 mm < 23 mm > 25 mm > 25 mm < 23 mm

Decaloba PassifloraFeuillet & MacDougal (2003) Astrophea Deidamioides

Killip (1938), Escobar (1988), MacDougal (1994) Astrophea Decaloba Psilanthus Passiflora Distephana Dysosmia Tacsonia Manicata TryphostemmatoidesSpecies/accession number n= 3/4 n= 17/31 n= 1/1 n= 19/44 n= 1/2 n= 1/2 n= 16/36 n= 1/1 n= 2/2Main pollinators medium (honey) bees small to large bees - wasps sword-billed large (carpenter) bees hummingbirds bees sword-billed hummingbirds bees

hummingbird hummingbirdChromosome number 2n = 24 2n = 12, 22, 24, 36 2n = 12 2n = 18 2n = 18 2n = 18, 20, 22 2n = 18 2n = 18 2n = 12

Altitudinal range 1.000 - 2.200 m 60 - 2.700 m 2.600 - 3.200 20 - 2.400 m 50 - 1.200 m 30 - 1.200 m 2.100 - 3.700 m 1.900 - 2.500 m 50 - 2.000 m

Descriptors

Habit tree vine vine vine vine vine vine vine vineStem section irregular terete/angular angular terete/angular/winged terete terete terete/angular angular tereteSecondery xylema present absent absent absent absent absent absent absent absentTendril position absent axillary axillary axillary axillary axillary axillary axillary axillary and peduncleStipule short triangular setaceous/linear/foliaceous-aristate setaceous setaceous/linear/foliaceous-aristate linear foliaceous-aristate setaceous/linear/foliaceous-aristate foliaceous-aristate setaceousStipule nectaries (conspicuous) absent absent absent absent/present (P. maliformis ) present present absent absent absentLeaf lobation one three three one/three/more three three one/three three oneLeaf base cuneate-rounded cuneate-rounded/cordate/ cordate cuneate-rounded/cordate cordate cordate cuneate-rounded/cordate cordate rounded

peltate (P. coriacea and P. guatemalensis )Leaf apex obtuse/acute rounded/obtuse/acute acute/very acute rounded to very acute acute acute acute/very acute obtuse to very acute retuseLeaf margin entire entire/serrate (P. adenopoda ) entire entire/serrate serrate serrate serrate serrate entireLaminar nectaries absent present/absent (five species)b present absent absent absent absent absent absentLeaf margin nectaries (conspicuous) absent absent/sinus/ absent absent/sinus sinus absent absent/sinus (P. jardinensis ) absent leaf base

all margin (P. adenopoda ) lateral lobe (P. edulis f. edulis )Petiolar nectaries absent absent/orbicular (P. adenopoda )/ absent all except auriculate orbicular absent linear/orbicular linear absent

cylindrical (P. coriaceae and P. suberosa )/auriculate (P. auriculata )

Dorsal nectaries present absent absent absent absent absent absent absent absentPeduncle branching present absent/present (P. sexflora ) absent absent absent absent absent absent presentBract shape linear setaceous/linear/ linear foliaceous linear pinnatisect foliaceous foliaceous linear

foliaceous (P. adenopoda , P. guatemalensis )/absent (four species)*

Nectary on bract absent absent absent absent/present present absent absent absent absentFlower orientation erect erect/intermediate/pendular pendular erect/intermediate/pendular erect erect erect/intermediate/pendular erect erectCorolla shape reflex intermediate/campanulate campanulate intermediate/reflex reflex intermediate/reflex campanulate/intermediate/reflex reflex reflexDominant corolla color white white red (pink) white/red red white red (including pink or orange) red whiteCorona type filamentous filamentous filamentous filamentous filamentous filamentous tuberculous/ filamentous filamentousCorona filaments sickle-sword-shaped linear linear linear linear linear filamentous, linear linear linear

free free free free fused at base free free free freeNumber of corona series uniseriate biseriate/ uniseriate pentaseriate biseriate 3- to 5-seriate uni- to 5-seriate pentaseriate biseriate

uniseriate (P. guatemalensis and P. adenopoda )/triseriate (P. filipes and P. magadalenae )

Corona color-clear (longest row) white white white red white white/purple purple whiteCorona color-darkest (longest row) yellow white/purple/ white purple red purple purple purple white

yellow (P. guatemalensis ) white (P. guazumaefolia ) white (P. jardinensis )Petals present present/ present present present present present present present

absent (P. coriaceae and P. suberosa )Sepal awn absent absent/present absent present present present present present absentNectar chamber ring present absent absent present present present present present absentHypanthium campanulate flat tubular campalunate tubular campanulate tubular tubular flatLimen present present absent present present present present present presentOvary shape tricostate globose globose globose globose globose globose globose globoseFruit shape globose globose/ elongate globose/ globose globose elongate/ elongate globose

elongate (P. rubra and P. capsularis ) elongate (P. quadrangularis ) globose (P. pinnatistipula )

Categorized (from quantitatives traits)Stem diameter ≥ 120 mm ≤ 20 mm ≤ 20 mm ≥ 30 mm ≥ 30 mm ≤ 20 mm ≥ 30 mm ≥ 30 mm ≤ 20 mm

< 60 mm < 60 mm < 60 mm < 60 mmLeaf length > 215 mm < 190 mm < 190 mm < 190 mm < 190 mm < 190 mm < 190 mm < 190 mm < 190 mm

< 190 mm (P. sphaerocarpa )Sepal length ≤ 30 mm ≤ 30 mm > 30 mm > 30 mm > 30 mm ≤ 30 mm > 30 mm > 30 mm ≤ 30 mm

≤ 30 mm (six species)ª ≤ 30 mm (P. luzmarina and P. mathewsii ) Androgynophore length < 23 mm < 23 mm > 25 mm < 23 mm > 25 mm < 23 mm > 25 mm > 25 mm < 23 mm

Passiflora

Feuillet & MacDougal (2003) Astrophea Deidamioides

Killip (1938), Escobar (1988), MacDougal (1994) Astrophea Decaloba Psilanthus Passiflora Distephana Dysosmia Tacsonia Manicata TryphostemmatoidesSpecies/accession number n= 3/4 n= 17/31 n= 1/1 n= 19/44 n= 1/2 n= 1/2 n= 16/36 n= 1/1 n= 2/2Main pollinators medium (honey) bees small to large bees - wasps sword-billed large (carpenter) bees hummingbirds bees sword-billed hummingbirds bees

hummingbird hummingbirdChromosome number

Altitudinal range 1.000 - 2.200 m 60 - 2.700 m 2.600 - 3.200 20 - 2.400 m 50 - 1.200 m 30 - 1.200 m 2.100 - 3.700 m 1.900 - 2.500 m 50 - 2.000 m

Descriptors

Habit tree vine vine vine vine vine vine vine vineStem section irregular terete/angular angular terete/angular/winged terete terete terete/angular angular tereteSecondery xylema present absent absent absent absent absent absent absent absentTendril position absent axillary axillary axillary axillary axillary axillary axillary axillary and peduncleStipule short triangular setaceous/linear/foliaceous-aristate setaceous setaceous/linear/foliaceous-aristate linear foliaceous-aristate setaceous/linear/foliaceous-aristate foliaceous-aristate setaceousStipule nectaries (conspicuous) absent absent absentLeaf lobation one three three one/three/more three three one/three three oneLeaf base cuneate-rounded cuneate-rounded/cordate/ cordate cuneate-rounded/cordate cordate cordate cuneate-rounded/cordate cordate rounded

Leaf apex obtuse/acute rounded/obtuse/acute acute/very acute rounded to very acute acute acute acute/very acute obtuse to very acute retuseLeaf margin entireLaminar nectaries absent present/absent (five species)b present absent absent absent absent absent absentLeaf margin nectaries (conspicuous) absent absent/sinus/ absent absent/sinus sinus absent

Petiolar nectaries absent

Dorsal nectaries present absent absent absent absent absent absent absent absentPeduncle branching presentBract shape linear setaceous/linear/ linear foliaceous linear pinnatisect foliaceous foliaceous linear

/absent (four species)*Nectary on bract absent absent absent absent/present present absent absent absent absentFlower orientation erect erect/intermediate/pendular pendular erect/intermediate/pendular erect erect erect/intermediate/pendular erect erectCorolla shape reflex intermediate/campanulate campanulate intermediate/reflex reflex intermediate/reflex campanulate/intermediate/reflex reflex reflexDominant corolla color white white red (pink) white/red red white red (including pink or orange) red whiteCorona type filamentous filamentous filamentous filamentous filamentous filamentous tuberculous/ filamentous filamentousCorona filaments sickle-sword-shaped linear linear linear linear linear filamentous, linear linear linear

free free free free fused at base free free free freeNumber of corona series uniseriate biseriate/ uniseriate pentaseriate biseriate 3- to 5-seriate uni- to 5-seriate pentaseriate biseriate

Corona color-clear (longest row) white white white red white white/purple purple whiteCorona color-darkest (longest row) yellow white/purple/ white purple red purple purple purple white

Petals present present/ present present present present present present present

Sepal awn absent absent/present absent present present present present present absentNectar chamber ring present absent absent present present present present present absentHypanthium campanulate flat tubular campalunate tubular campanulate tubular tubular flatLimen present present absent present present present present present presentOvary shape tricostate globose globose globose globose globose globose globose globoseFruit shape globose globose/ elongate globose/ globose globose elongate/ elongate globose

Categorized (from quantitatives traits)Stem diameter ≥ 120 mm ≤ 20 mm ≤ 20 mm ≥ 30 mm ≥ 30 mm ≤ 20 mm ≥ 30 mm ≥ 30 mm ≤ 20 mm

< 60 mm < 60 mm < 60 mm < 60 mmLeaf length > 215 mm < 190 mm < 190 mm < 190 mm < 190 mm < 190 mm < 190 mm < 190 mm < 190 mm

Sepal length ≤ 30 mm ≤ 30 mm > 30 mm > 30 mm > 30 mm ≤ 30 mm > 30 mm > 30 mm ≤ 30 mm

Androgynophore length < 23 mm < 23 mm > 25 mm < 23 mm > 25 mm < 23 mm > 25 mm > 25 mm < 23 mm

32 Discriminant qualitative characters between subgenera

e. Qualitative data dendrogram (nNeighbour Joining, 32 characters)

n = 9n = 6

n = 12

Distances Sokal & Michener

n = 10

n = ??

n = 6

Conclusions

The new descriptors list has permitted to rank species together with taxonomy in a coherent way. However, there are some exceptions.

The analysis is coherent with Killip's classification (1938) at the subgenus level.

Descriptors related to the flower permit a higher discrimination between subgenera and species.

III. Genetic diversity (CAPS – DNAcp/mt)

Taxon sampling

• 213 individuals• 151 species • 15 subgenera (Killip, 1938)

Origin• Collections in Colombia – 75 %• Passion Fruit National Collections – Blois (France) – 22 %• Royal Botanical Garden – Meise (Belgium) – 2 %• Cameroon and Gabon - 1% (outgroup)

Tecnic – Markers RFLP-PCR (2 couple of primers / 6 enzymes)

Nuclear Sequences

ncpGS

- 91 species

- 17 subgenera (Killip, 1938)

Yockteng & Nadot (2004)DECALO

BA

ASTROPHEA

PASSIF

LORA

Outgroup

Previous studies

Hansen et al. (2006)

Chloroplas Sequences TrnL / TrnT

- 57 species

- 16 subgenera (Killip, 1938)

DECALOBA

ASTROPHEA

PASSIF

LORA

outgroup

Previous studies

Lad

der

P. o

erst

edii

P. m

alifo

rmis

P. s

mith

iiP

. le

hman

nii

P. a

lata

P. a

mbi

gua

P. c

inci

nnat

aP

. gar

ckei

P. p

open

ovii

P. t

risul

caP

. cae

rule

aP

. qua

dran

gula

risP

. citr

ifolia

P. c

oria

ceae

P. f

ilipe

sP

. ery

trop

hylla

P. s

uber

osa

P. c

unea

taP

. mag

dale

nae

P.

linea

ristip

ula

P. a

ntio

quie

nsis

P. p

arrit

aeP

. tric

uspi

sP

. aln

ifolia

P.

luzm

arin

aP

. fle

xipe

sP

. mix

taP

. tar

min

ana

Lad

der

TS1TS1--TS2 (TS2 (HpaHpaIIII))

N1B1N1B1--N1C (N1C (HpaHpaIIII))

Chloroplast

Mitochondria

Cytoplasmatic genome inter-specific variation

280 haplotypes

372 haplotypes

Chlorotypes are easier to interpret than mitotypes

Primers Hinf I Rsa I Hpa II Hae III Hha I

Taq I

PC1 – PC2

Decaloba two group - Adenia - Barteria -Smeathmannia - other subgenera

Impossible to interpret

Decaloba three groups - other subgenera

Decaloba - Adenia -Smeathmannia - other subgenera

Decaloba three groups - Passiflora two groups - Tacsonia two group - other subgenera

Decaloba - other subgenera

TS1 – TS2 Decaloba two groups - Passiflora four groups - (Barteria - Smeathmannia) - other subgenera

Decaloba three groups - Passiflora four groups - Astrophea - other subgenera

Decaloba two groups - Passiflora three groups - Adenia - (Barteria - Smeathmannia) - other subgenera

Decaloba three groups - Passiflora two groups - other subgenera

Decaloba two groups - Passiflora two groups - other subgenera

Decaloba three groups - Passiflora two groups - Distephana - Astrophea - Adenia - (Barteria - Smeathmannia) - other subgenera

N41 – N42

Decaloba two groups - Passiflora - Astrophea - (Barteria - Smeathmannia) - other subgenera

Decaloba three groups - Passiflora two groups - Adenia - other subgenera

Decaloba two groups Passiflora four groups - Tacsonia two groups - Barteria two groups - other subgenera

Decaloba two - Passiflora five groups - Adenia - other subgenera

Decaloba two groups - Passiflora three groups - Barteria two groups - other subgenera

Impossible to interpret

N1B – N1C Decaloba three groups - Passiflora two groups - Tacsonia two groups - (Barteria - Smeathmannia) - Adenia - other subgenera

Decaloba two groups - Passiflora two groups - Tacsonia - (Barteria - Smeathmannia) - Adenia - other subgenera

Decaloba two groups - Passiflora two groups - Tacsonia two groups - (Barteria - Smeathmannia) - other subgenera

Decaloba two groups - Passiflora two groups - Tacsonia three two - other subgenera

Decaloba two groups - Passiflora two groups - Tacsonia two groups - Murucuja - Adenia - other subgenera

Decaloba two groups - Tacsonia - Barteria two groups - Adenia -

Observed variation per primer-enzyme combination (haplotypes)

Mit

och

on

dri

al

C

hlo

rop

last

ic

AFTD of Chloroplastic data

OUTGROUP

Chloroplast DNAAstropheaTryphostemmatoides

- Three big groups

Decaloba – much better separated and structured

Passiflora - little structure

Astrophea

- Genus monophyly ?

- Tryphostemmatoides

NJ, distance de Sokal & Michener

Mitochondrial DNA

Separation of the Decaloba group, better structured

AstropheaTryphostemmatoides

Strong approximation of Astrophea and Tryphostemmatoides to Passiflora

Tacsonia

Little cohesion with morphology at the section and series levels.

Tacsonia

Geographic structuring by:

- altitude range (Tacsonia, stip. ren.)

- region (Tacsonia, Ecuador/Colombia).

Genus monophyly ?

Conclusions

• Divergence between morphologic, chloroplastic and mitochondrial data explains plastid transmission modes:

Chloroplastic genome transmission is basically paternal or biparental in the Passiflora group, and maternal in the mitochondiral genome.

• The doubt on whether the Passiflora genus is not monophyletic imposes a re-examination of the generic classification of Passifloraceae.

• The data set must be interpreted taking into account the geological context of tropical flora separation and the formation of the Andean mountain range.

• RETICULATE EVOLUTION

Future prospects

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.

Passion fruits and genetic resourcesExplore the possibility of interspecific hybridization to compensate for the deficiencies of the main cultivated species.

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.

Thanks!