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A survey of Phytophthora and
  Pythium associated to export
       crops in Guatemala




P. Abad-Campos1, A. Pérez-Sierra1, L.A. Álvarez1, J. Armengol1, R. López-
   Pineda2, A. Sánchez-Pérez2, E. Rodríguez-Quezada3 and G. Álvarez-
                              Valenzuela2
      1 Instituto Agroforestal Mediterráneo, Universidad Politécnica de Valencia (Spain)
                2 Facultadde Agronomía, Universidad San Carlos de Guatemala
                3 Laboratorio de Diagnóstico Fitosanitario, M.A.G.A. (Guatemala)
A Survey of Phytophthora and Pythium associated to
                 export crops in Guatemala


•   Located within the tropics and with elevations
    ranging between sea level and more than
    4.000 m, Guatemala experiences a diversity of
    climates.
•   Guatemala has mainly agricultural economy.
•   Subsistence crop production includes corn, dry
    beans, rice,corn, and fruits and vegetables.
•   Agriculture accounts for 23% of the GNP and
    75% of the country’s exports.
•   Nontraditional agricultural exports have greatly
    increased in recent years; such products
    include: banana, brassicas, beans, cacao,
    coffee, cucumbers, melon, mango, onion,
    papaya, pineapple, and ornamental plants.
A Survey of Phytophthora and Pythium associated to
                 export crops in Guatemala

•   Diseases caused by Oomycetes have been
    detected with increasing incidence in several
    crops.
•   Surveys of ornamental, vegetable and fruit
    crops were carried out during 2007 mainly in
    Central Region of Guatemala (Chimaltenango,
    Sacatepéquez and Guatemala Departments).
•   Samples were collected during the humid
    period of the year (March- November).
A Survey of Phytophthora and Pythium associated to
                 export crops in Guatemala

•   Sacatepéquez: Camino de San Pedro, Chitol, Cruz Verde, El Pilar, Loma
    Alta, Los Guamuches, Santa Fe Ocaña, Sajcavilla, San Juan, Zet
•   Chimaltenango: Comalapa, El Tejar, Parramos, Patzicia, Patzún, Rincón,
    Saragoza, San Andrés Itzapa, San Martín Jilotepeque, Sumpango
•   Guatemala: Villa Canales, Villa Nueva, Guatemala City
•   Alta Verapaz: Chirijuyú
•   Baja Verapaz: Chilasco
•   Jalapa: Tucurú
•   Petén: Flores
•   Huehuetenango: Jacaltenango, El Piol San Sebastián
•   San Marcos: Malacatán
•   Quezaltenango: Almolonga
•   Escuintlá: Escuintlá, Finca Sabana Grande
•   Santa Rosa: Pueblo Nuevo Viñas, Guazacapán
•   Sololá: Chiarmira, Chacayá
•   Suchitepéquez: Bulbuyá
A Survey of Phytophthora and Pythium associated to export crops
                         in Guatemala




El Jocotillo - Villa Canales, Guatemala (800 m)
A Survey of Phytophthora and Pythium associated to export crops
                         in Guatemala




 San Juan, Sacatepéquez (1700 m)
A Survey of Phytophthora and Pythium associated to export crops
                         in Guatemala




 Comunidad de Zet, Sacatepéquez (1787 m)
A Survey of Phytophthora and Pythium associated to export crops
                         in Guatemala




 Sumpango, Sacatepéquez (1791m)
A Survey of Phytophthora and Pythium associated to export crops
                         in Guatemala




Patzicia, Chimaltenango (2083 m)
A Survey of Phytophthora and Pythium associated to export crops
                         in Guatemala




  El Rincón, Chimaltenango (2.207 m)
A Survey of Phytophthora and Pythium associated to
                export crops in Guatemala

SAMPLED CROPS
• Leguminoses: beans and snow pea
  pod (Pisum sativum var. Saccharatum)
• Vegetables: brassicas, carrot,
  cucumber, lettuce, onion, pepper,
  potato, and tomato
• Fruits: avocado, lemon, pineapple, and
  strawberry
• Cacao and coffee trees
• Ornamentals: asparagus, carnation,
  chrysanthemum, gladiolus, iris,
  leatherleaf fern, lilium, peperomia,
  ponytail grass, pothos, rose, sheflera
• Gold tree or ‘Palo blanco’ (Tabebuia
  donnell-smithii)                         Gold tree
A Survey of Phytophthora and Pythium associated to
             export crops in Guatemala
A Survey of Phytophthora and Pythium associated to
             export crops in Guatemala


                                         Pothos




         Rose
                                     Carnation
A Survey of Phytophthora and Pythium associated to
             export crops in Guatemala




         Chrysanthemum            Lilium
A Survey of Phytophthora and Pythium associated to
              export crops in Guatemala
Leatherleaf fern                        Ornamental asparagus
A Survey of Phytophthora and Pythium associated to
             export crops in Guatemala
A Survey of Phytophthora and Pythium associated to
             export crops in Guatemala
A Survey of Phytophthora and Pythium associated to
             export crops in Guatemala
A Survey of Phytophthora and Pythium associated to
             export crops in Guatemala
MATERIALS AND METHODS

ISOLATIONS
• Isolations from lesions in roots, stems and leaves of
  symptomatic plants were made by direct planting on
  selective media for Oomycetes (PARBH and PARB)
• Cacao pods were previously incubated in humid
  chamber at room temperature
• Soil samples were tested using apple baits
• Pure cultures were obtained by transferring single
  hyphal tips to PDA
MATERIALS AND METHODS


CHARACTERIZATION
• Cultural pattern on Potato dextrose agar
• Morphology:

   - Asexual characteristics:
       •Sporangia: nonpapillate, semipapillate, or papillate
       •Persistence or caducity of sporangia
       •Sporangium shape and size
       •Sporangium length – breadth (L:B) ratio
       •Sporangiophore: distribution
       •Hyphal swellings: single, catenulate or clustered
       •Chlamydospores: terminal, lateral, intercalary or sessile
MATERIALS AND METHODS

CHARACTERIZATION
  - Sexual characteristics: pairing of isolates on CMA and V8
      Isolates paired with A1 and A2 mating types of P. cryptogea
      •Oospore: diameter and wall tickness                Dual cultures

      •Oospore: plerotic or aplerotic
      •Homothallic or heterothallic


  - Growth cardinal temperatures: T °C minimum and maximum


MOLECULAR IDENTIFICATION
- Amplification of ITS-5.8S rDNA using ITS4 and ITS6 primers
        (White et al., 1990; Cooke et al., 2000)
- Sequencing and alignment with sequences available in the EMBL/GenBank
RESULTS


Six Phytophthora species were identified
  associated to different crops:
P. capsici: pepper and pothos
P. cinnamomi: avocado
P. citrophthora: pothos
P. palmivora: avocado, cacao, and Persian
  lemon
P. nicotianae: ‘Palo blanco’ tree and Persian
  lemon
P. tropicalis: carnation, peperomia, pepper,
  pothos, and rose
RESULTS
P. capsici and P. tropicalis

Long-pedicellate caducous sporangia
Papillate and semipapillate sporangia
Most of them presented a L:B ratio > 1.8:1
Absence of chlamydospores
Heterothallism; present both mating types
Some isolates grew well at 35ºC and others had limited growth or
were inhibited at this temperature
ITS sequence was not useful to differentiate both species




  Papillate (left) and
  semipapillate (right)
  sporangia
                           P. capsici            P. tropicalis
RESULTS
P. cinnamomi




   Non papillate sporangium   Colony pattern on PDA




      Oogonium                Chlamydospores
RESULTS

P. palmivora




                    Sporangia




         Oogonium        Colony pattern on PDA
RESULTS

 P. citrophthora             P. nicotianae




     Sporangia          Sporangia          Oogonium




Colony pattern on PDA   Colony pattern on PDA
RESULTS

Several Pythium species were isolated from
different crops
   Identification was based on ITS sequence

•Py. cucurbitacearum: broccoli, carnation,
onion, papaya, pepper, Persian lemon, and
ponytail grass
•Py. splendens: avocado, coffee tree, and
ornamental asparagus
•Py. sylvaticum: ornamental asparagus
•Py. ultimum: iris, lettuce, potato, and squash
•Pythium sp.: avocado, chrysanthemum, coffee
tree, leatherleaf fern, lilium, sheflera, and snow
pea pod
We acknowledge the financial support of ‘Agencia
 Española de Cooperación Internacional para el
        Desarrollo’ (AECID, A/6711/06)

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O15 Abad

  • 1. A survey of Phytophthora and Pythium associated to export crops in Guatemala P. Abad-Campos1, A. Pérez-Sierra1, L.A. Álvarez1, J. Armengol1, R. López- Pineda2, A. Sánchez-Pérez2, E. Rodríguez-Quezada3 and G. Álvarez- Valenzuela2 1 Instituto Agroforestal Mediterráneo, Universidad Politécnica de Valencia (Spain) 2 Facultadde Agronomía, Universidad San Carlos de Guatemala 3 Laboratorio de Diagnóstico Fitosanitario, M.A.G.A. (Guatemala)
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  • 3. A Survey of Phytophthora and Pythium associated to export crops in Guatemala • Located within the tropics and with elevations ranging between sea level and more than 4.000 m, Guatemala experiences a diversity of climates. • Guatemala has mainly agricultural economy. • Subsistence crop production includes corn, dry beans, rice,corn, and fruits and vegetables. • Agriculture accounts for 23% of the GNP and 75% of the country’s exports. • Nontraditional agricultural exports have greatly increased in recent years; such products include: banana, brassicas, beans, cacao, coffee, cucumbers, melon, mango, onion, papaya, pineapple, and ornamental plants.
  • 4. A Survey of Phytophthora and Pythium associated to export crops in Guatemala • Diseases caused by Oomycetes have been detected with increasing incidence in several crops. • Surveys of ornamental, vegetable and fruit crops were carried out during 2007 mainly in Central Region of Guatemala (Chimaltenango, Sacatepéquez and Guatemala Departments). • Samples were collected during the humid period of the year (March- November).
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  • 6. A Survey of Phytophthora and Pythium associated to export crops in Guatemala • Sacatepéquez: Camino de San Pedro, Chitol, Cruz Verde, El Pilar, Loma Alta, Los Guamuches, Santa Fe Ocaña, Sajcavilla, San Juan, Zet • Chimaltenango: Comalapa, El Tejar, Parramos, Patzicia, Patzún, Rincón, Saragoza, San Andrés Itzapa, San Martín Jilotepeque, Sumpango • Guatemala: Villa Canales, Villa Nueva, Guatemala City • Alta Verapaz: Chirijuyú • Baja Verapaz: Chilasco • Jalapa: Tucurú • Petén: Flores • Huehuetenango: Jacaltenango, El Piol San Sebastián • San Marcos: Malacatán • Quezaltenango: Almolonga • Escuintlá: Escuintlá, Finca Sabana Grande • Santa Rosa: Pueblo Nuevo Viñas, Guazacapán • Sololá: Chiarmira, Chacayá • Suchitepéquez: Bulbuyá
  • 7. A Survey of Phytophthora and Pythium associated to export crops in Guatemala El Jocotillo - Villa Canales, Guatemala (800 m)
  • 8. A Survey of Phytophthora and Pythium associated to export crops in Guatemala San Juan, Sacatepéquez (1700 m)
  • 9. A Survey of Phytophthora and Pythium associated to export crops in Guatemala Comunidad de Zet, Sacatepéquez (1787 m)
  • 10. A Survey of Phytophthora and Pythium associated to export crops in Guatemala Sumpango, Sacatepéquez (1791m)
  • 11. A Survey of Phytophthora and Pythium associated to export crops in Guatemala Patzicia, Chimaltenango (2083 m)
  • 12. A Survey of Phytophthora and Pythium associated to export crops in Guatemala El Rincón, Chimaltenango (2.207 m)
  • 13. A Survey of Phytophthora and Pythium associated to export crops in Guatemala SAMPLED CROPS • Leguminoses: beans and snow pea pod (Pisum sativum var. Saccharatum) • Vegetables: brassicas, carrot, cucumber, lettuce, onion, pepper, potato, and tomato • Fruits: avocado, lemon, pineapple, and strawberry • Cacao and coffee trees • Ornamentals: asparagus, carnation, chrysanthemum, gladiolus, iris, leatherleaf fern, lilium, peperomia, ponytail grass, pothos, rose, sheflera • Gold tree or ‘Palo blanco’ (Tabebuia donnell-smithii) Gold tree
  • 14. A Survey of Phytophthora and Pythium associated to export crops in Guatemala
  • 15. A Survey of Phytophthora and Pythium associated to export crops in Guatemala Pothos Rose Carnation
  • 16. A Survey of Phytophthora and Pythium associated to export crops in Guatemala Chrysanthemum Lilium
  • 17. A Survey of Phytophthora and Pythium associated to export crops in Guatemala Leatherleaf fern Ornamental asparagus
  • 18. A Survey of Phytophthora and Pythium associated to export crops in Guatemala
  • 19. A Survey of Phytophthora and Pythium associated to export crops in Guatemala
  • 20. A Survey of Phytophthora and Pythium associated to export crops in Guatemala
  • 21. A Survey of Phytophthora and Pythium associated to export crops in Guatemala
  • 22. MATERIALS AND METHODS ISOLATIONS • Isolations from lesions in roots, stems and leaves of symptomatic plants were made by direct planting on selective media for Oomycetes (PARBH and PARB) • Cacao pods were previously incubated in humid chamber at room temperature • Soil samples were tested using apple baits • Pure cultures were obtained by transferring single hyphal tips to PDA
  • 23. MATERIALS AND METHODS CHARACTERIZATION • Cultural pattern on Potato dextrose agar • Morphology: - Asexual characteristics: •Sporangia: nonpapillate, semipapillate, or papillate •Persistence or caducity of sporangia •Sporangium shape and size •Sporangium length – breadth (L:B) ratio •Sporangiophore: distribution •Hyphal swellings: single, catenulate or clustered •Chlamydospores: terminal, lateral, intercalary or sessile
  • 24. MATERIALS AND METHODS CHARACTERIZATION - Sexual characteristics: pairing of isolates on CMA and V8 Isolates paired with A1 and A2 mating types of P. cryptogea •Oospore: diameter and wall tickness Dual cultures •Oospore: plerotic or aplerotic •Homothallic or heterothallic - Growth cardinal temperatures: T °C minimum and maximum MOLECULAR IDENTIFICATION - Amplification of ITS-5.8S rDNA using ITS4 and ITS6 primers (White et al., 1990; Cooke et al., 2000) - Sequencing and alignment with sequences available in the EMBL/GenBank
  • 25. RESULTS Six Phytophthora species were identified associated to different crops: P. capsici: pepper and pothos P. cinnamomi: avocado P. citrophthora: pothos P. palmivora: avocado, cacao, and Persian lemon P. nicotianae: ‘Palo blanco’ tree and Persian lemon P. tropicalis: carnation, peperomia, pepper, pothos, and rose
  • 26. RESULTS P. capsici and P. tropicalis Long-pedicellate caducous sporangia Papillate and semipapillate sporangia Most of them presented a L:B ratio > 1.8:1 Absence of chlamydospores Heterothallism; present both mating types Some isolates grew well at 35ºC and others had limited growth or were inhibited at this temperature ITS sequence was not useful to differentiate both species Papillate (left) and semipapillate (right) sporangia P. capsici P. tropicalis
  • 27. RESULTS P. cinnamomi Non papillate sporangium Colony pattern on PDA Oogonium Chlamydospores
  • 28. RESULTS P. palmivora Sporangia Oogonium Colony pattern on PDA
  • 29. RESULTS P. citrophthora P. nicotianae Sporangia Sporangia Oogonium Colony pattern on PDA Colony pattern on PDA
  • 30. RESULTS Several Pythium species were isolated from different crops Identification was based on ITS sequence •Py. cucurbitacearum: broccoli, carnation, onion, papaya, pepper, Persian lemon, and ponytail grass •Py. splendens: avocado, coffee tree, and ornamental asparagus •Py. sylvaticum: ornamental asparagus •Py. ultimum: iris, lettuce, potato, and squash •Pythium sp.: avocado, chrysanthemum, coffee tree, leatherleaf fern, lilium, sheflera, and snow pea pod
  • 31. We acknowledge the financial support of ‘Agencia Española de Cooperación Internacional para el Desarrollo’ (AECID, A/6711/06)