, Volume 169, Issue 1, pp 1–17 | Cite as

Platanus × acerifolia genotypes surviving to inoculation with Ceratocystis platani (the agent of canker stain): first screening and molecular characterization

  • M. PilottiEmail author
  • A. Brunetti
  • L. Tizzani
  • O. Marani


Canker stain, caused by the fungus Ceratocystis platani, is a destructive disease in Platanus spp. It has been recently proved that resistant accessions can be produced and grown in Europe. However, additional resistant genotypes are still needed in order to avoid the onset of virulent pathogen strains favoured by the selection pressures exerted by genetically homogeneous resistant plane tree plantings. In this study we present the results of two parallel experiments performed on 975 accessions of P. × acerifolia seedlings and P. × acerifolia clones derived by cutting propagation from mature trees grown in the urban environments. The selection process was based on inoculation with C. platani and yielded 13 accessions that showed different types of resistant reactions and survived in a stable manner thoughout the period of observation. Selected accessions were characterised by sequencing the rDNA-ITS region and by developing PCR procedures capable of detecting P. orientalis and P. occidentalis LEAFY homoeologues. These molecular analyses enabled us to confirm the identification of the species, its hybrid origin and to assess an evident genetic variability among the accessions, which therefore have to be considered as different genotypes.


Platanus Canker stain Ceratocystis platani Resistance ITS region LEAFY gene 



This ten-year long research could not have been realised without the vital support of pubblic Institutions and the help and engagement of many persons. The research project was funded over the years by Servizio Giardini—Comune di Roma. In particular we are grateful to Dr. Dario Esposito, Dr. Stefano Mastrangelo and Dr. Angelico Bonuccelli. Thanks also to Mr. Augusto Burini and Mr. Francesco Messina for their valuable contribution in supporting the research projects and to all the staff of Servizio Giardini that helped us in collecting and taking care of plant material. Molecular investigation was supported by MIPAF—Ministero delle Politiche Agricole e Forestali, project: “Proteine e geni per la protezione delle piante dagli stress biotici e abiotici, Proteo Stress—Genomica e Proteomica per lo studio della risposta di resistenza in specie arbore, ciliegio e platano”. We express our thanks to the coordinator of the project, Prof Felice Cervone (University La Sapienza, Rome). We would also like to mention Prof Paolo Capretti (University of Florence), Dr. Guido W. Grimm (Eberhard-Karls-University) and Dr. Angelo Porta-Puglia for their useful comments on the manuscript. Finally, we express our gratefulness to Dr. Andrè Vigouroux (INRA, France) whose research experience on the subject inspired us with the selection procedures in this study.

Supplementary material

10681_2009_9884_MOESM1_ESM.doc (130 kb)
Supplementary material (DOC 130 kb)


  1. Annesi T, Motta E, Pilotti M (2003) Diagnostic protocols for regulated pests: Ceratocystis fimbriata f. sp. platani. EPPO Bull 33:249–255. doi: 10.1046/j.1365-2338.2003.00640.x CrossRefGoogle Scholar
  2. Anselmi N, Cardin L, Nicolotti G (1994) Plane decline in European and Mediterranean countries: associated pests and their interactions. EPPO Bull 24:159–171. doi: 10.1111/j.1365-2338.1994.tb01058.x CrossRefGoogle Scholar
  3. Baker CJ, Harrington TC, Krauss U, Alfenas AC (2003) Genetic variability and host specialization in the latin American clade of Ceratocystis fimbriata. Phytopathol 93:1274–1284. doi: 10.1094/PHYTO.2003.93.10.1274 CrossRefGoogle Scholar
  4. Besnard G, Tagmount A, Baradat P, Vigouroux A, Bervillé A (2002) Molecular approach of genetic affinities between wild and ornamental Platanus. Euphytica 126:401–412. doi: 10.1023/A:1019912425987 CrossRefGoogle Scholar
  5. Britton KO, Leininger R, Chang CJ, Harrington TC (1998) Association of Xilella fastidiosa, Ceratocystis fimbriata f. platani and Botryosphaeria rhodina with declining sycamore plantations in the south-eastern USA. In: Proceedings of ICPP 98, Edinburgh, Poster 3.7.50Google Scholar
  6. Buckler ES, Ippolito A, Holtsford TP (1997) The evolution of ribosomal DNA: divergent paralogues and phylogenetic implications. Genetics 145:821–832PubMedGoogle Scholar
  7. Crane PR, Pedersen KR, Friis EM, Drinnan AN (1993) Early cretaceous (early to middle Albian) platanoid inflorescence associated with sapindopsis leaves from the Potomac group of group of eastern North America. Syst Bot 18:328–344. doi: 10.2307/2419407 CrossRefGoogle Scholar
  8. Cristinzio M, Marziano F, Verneau R (1973) La moria del platano in Campania. Riv Pat Veg S IV 9:189–214Google Scholar
  9. Efron B (1982) The Jackknife, the bootstrap and other resembling plans. SIAM, PhiladelphiaGoogle Scholar
  10. Engelbrecht CJB, Harrington TC (2005) Intersterility, morphology and taxonomy of Ceratocystis fimbriata on sweet potato, cacao and sycamore. Mycol 97:57–69. doi: 10.3852/mycologia.97.1.57 CrossRefGoogle Scholar
  11. Engelbrecht CJB, Harrington TC, Steimel J, Capretti P (2004) Genetic variation in eastern north American and putatively introduced populations of Ceratocystis fimbriata f. platani. Mol Ecol 13:2995–3005. doi: 10.1111/j.1365-294X.2004.02312.x PubMedCrossRefGoogle Scholar
  12. Feng Y, Oh SH, Manos PS (2005) Phylogeny and historical biogeography of the genus Platanus as inferred from nuclear and and chloroplast DNA. Syst Bot 30:786–799. doi: 10.1600/036364405775097851 CrossRefGoogle Scholar
  13. Göker M, Grimm GW (2008) General functions to transform associated data to host data and their use in phylogenetic inference from sequences with intra-individual variability. BMC Evol Biol 8:86. doi: 10.1186/1471-2148-8-86 PubMedCrossRefGoogle Scholar
  14. Granata G, Pennisi AM (1989) Estese morie di platani in forestazioni naturali causate da Ceratocystis fimbriata (Ell. et Halst.) Davidson f. platani Walter. Fitopatol 39:59–61Google Scholar
  15. Grimm GW, Denk T (2008) ITS Evolution in Platanus (Platanaceae): homoeologues, pseudogenes and ancient hybridization. Ann Bot (Lond) 101:403–419. doi: 10.1093/aob/mcm305 CrossRefGoogle Scholar
  16. Heath MC (2000) Hypersensitive response-related death. Plant Mol Biol 44:321–334. doi: 10.1023/A:1026592509060 PubMedCrossRefGoogle Scholar
  17. Henry A, Floods MG (1919) The hystory of the London Plane (Platanus acerifolia). Notes on the genus Platanus. Proc R Ir Acad [B] 35:9–28Google Scholar
  18. Jackson LWR, Sleeth B (1935) A new disease affecting Platanus orientalis in the Eastern United States. Phytopathol 25:22Google Scholar
  19. Jones JDG, Dangl JL (2006) The plant immune system. Nature 444:323–329PubMedCrossRefGoogle Scholar
  20. Kellog EA, Bennetzen JL (2004) The evolution of nuclear genome structure in seed plants. Am J Bot 91:1709–1725. doi: 10.3732/ajb.91.10.1709 CrossRefGoogle Scholar
  21. Kumar S, Tamura K, Nei M (2004) MEGA3: integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5:150–163. doi: 10.1093/bib/5.2.150 PubMedCrossRefGoogle Scholar
  22. Lam E, Kato N, Lawton M (2001) Programmed cell death, mitochondria and the plant hypersensitive response. Nature 411:848–853PubMedCrossRefGoogle Scholar
  23. McDowell JM, Woffenden BJ (2003) Plant disease resistance genes: recent insights and potential applications. Trends Biotech 21:178–183CrossRefGoogle Scholar
  24. Nixon KC, Poole JM (2003) Revision of the Mexican and Guatemala species of Platanus (Platanaceae). Lundellia 6:103–137Google Scholar
  25. Ocasio-Morales RG, Tsopelas P, Harrington TC (2007) Origin of Ceratocystis platani on native Platanus orientalis in Greece and its impact on natural forests. Plant Dis 91:901–904CrossRefGoogle Scholar
  26. Panconesi A (1999) Canker stain of plane trees: a serious danger to urban plantings. J Plant Pathol 81:3–15Google Scholar
  27. Pikaard CS (2001) Genomic change and gene silencing in polyploids. Trends Genet 17:675–677PubMedCrossRefGoogle Scholar
  28. Pilotti M (2002) Le avversità del platano. Fitopatol 52:9–24Google Scholar
  29. Pilotti M, Ponzio V, Motta E (2002a) Disorders of Platanus × acerifolia in Italy associated with Fusarium solani. For Pathol 32:249–264Google Scholar
  30. Pilotti M, Ponzio V, Coppola R, Motta E (2002b) Selezione per la resistenza al cancro colorato in Platanus × acerifolia. Fitopatol 52:28–37Google Scholar
  31. Pilotti M, Brunetti A, Gallelli A, Loreti S (2008) NPR1-like genes from cDNA of rosaceous trees: cloning strategy and genetic variation. Tree Genet Genomes 4:49–63CrossRefGoogle Scholar
  32. Ponchet J, Andrèoli C (1990) Compartmentalization and reactions in the host. In: Ponchet J (ed) Progress in EEC research on cypress disease. Commission of the European Communities, Brussels, pp 96–111Google Scholar
  33. Raddi P, Panconesi A (1981) Cypress canker disease in Italy: biology, control possibilities and genetic improvement for resistance. For Pathol 11:340–347Google Scholar
  34. Raddi P, Panconesi A (1984) Pathogenicity of some isolates of Seridium (Coryneum)cardinale, agent of cypress canker disease. For Pathol 14:348–354Google Scholar
  35. Saitou N, Nei M (1987) The neighbour-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425PubMedGoogle Scholar
  36. Santini A, Capretti P (2000) Analysis of the Italian population of Ceratocystis fimbriata f. sp. platani using RAPD and minisatellite markers. Plant Pathol 49:461–467CrossRefGoogle Scholar
  37. Santini A, Di Lonardo V (2000) Genetic variability of the ‘bark canker resistance’ character in several natural provenances of Cupressus sempervirens. For Pathol 30:87–96Google Scholar
  38. Santini A, La Porta N, Ghelardini L, Mittempergher L (2007) Breeding against Dutch elm disease adapted to the Mediterranean climate. Euphytica 163(1):45–56. doi: 10.1007/s10681-007-9573-5 CrossRefGoogle Scholar
  39. Smalley EB, Guries RP (1993) Breeding elms for resistance to Dutch elm disease. Annu Rev Phytopathol 31:325–352CrossRefGoogle Scholar
  40. Stanford AM, Harden R, Parks CR (2000) Phylogeny and biogeography of Juglans (Juglandaceae) based on matK and ITS sequence data. Am J Bot 87:872–882PubMedCrossRefGoogle Scholar
  41. Vigouroux A (1999) Main cryptogamic disorders of urban trees. Example of the plane tree with special emphasis on canker stain. Acta Hort 496:93–98Google Scholar
  42. Vigouroux A, Olivier R (2004) First hybrid plane trees to show resistance against canker stain (Ceratocystis fimbriata f. sp. platani). For Pathol 34:307–319Google Scholar
  43. Wendel JF (2000) Genome evolution in polyploids. Plant Mol Biol 42:225–249PubMedCrossRefGoogle Scholar
  44. White TJ, Bruns TD, Lee S, Taylor JW (1990) PCR protocols. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. Academic Press, San DiegoGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • M. Pilotti
    • 1
    Email author
  • A. Brunetti
    • 1
  • L. Tizzani
    • 1
  • O. Marani
    • 1
  1. 1.Centro di Ricerca per la Patologia VegetaleConsiglio per la Ricerca e la Sperimentazione in Agricoltura, CRA-PAVRomeItaly

Personalised recommendations