Australasian Plant Pathology

, Volume 40, Issue 5, pp 540–548 | Cite as

Distribution of Diplodia pinea and its genotypic diversity within asymptomatic Pinus patula trees

  • Wubetu Bihon
  • Treena Burgess
  • Bernard Slippers
  • Michael J. Wingfield
  • Brenda D. Wingfield
Article

Abstract

Diplodia pinea (= Sphaeropsis sapinea) is an endophytic fungus and opportunistic canker pathogen of Pinus spp. The diversity of this fungus has been studied at broad geographic scales, but little is known regarding its population structure at smaller spatial scales such as within a single tree. This is despite the importance that diversity in a single tree might hold for understanding the biology of the fungus, especially the role of the endophytic or asymptomatic phase in disease development. Moreover there was not information regarding the distribution of the fungus within healthy trees and its persistence. The genotypic diversity of these isolates was investigated using microsatellite markers. Five polymorphic markers were developed and these were used together with eight previously developed markers and vegetative compatibility tests to study the genotypic diversity of D. pinea isolates. In this study, D. pinea was isolated for the first time in the well structured stems of healthy P. patula trees along with branches and cones. From a total of 44 isolates collected from five trees, 39 microsatellite haplotypes and 32 vegetative compatibility groups (VCG’s) were identified. The results indicate high genotypic diversity of D. pinea within individual asymptomatic trees which will lead to disease outbreak when trees are physiologically stressed.

Keywords

Microsatellite markers Population diversity Vegetative compatibility groups Persistence 

References

  1. Blodgett JT, Kruger EL, Stanosz GR (1997) Effects of moderate water stress on disease development by Sphaeropsis sapinea on red pine. Phytopathology 87:422–428PubMedCrossRefGoogle Scholar
  2. Breuillin F, Dutech C, Robin C (2006) Genetic diversity of the chestnut blight fungus Cryphonectria parasitica in four French populations assessed by microsatellite markers. Mycol Res 110:288–296PubMedCrossRefGoogle Scholar
  3. Burgess T, Wingfield MJ (2001) Exotic pine forestry in the Southern Hemisphere: a brief history of establishment and quarantine practices. S Afr For J 192:79–84Google Scholar
  4. Burgess T, Wingfield BD, Wingfield MJ (2001a) Comparison of genotypic diversity in native and introduced populations of Sphaeropsis sapinea isolated from Pinus radiata. Mycol Res 105:1331–1339CrossRefGoogle Scholar
  5. Burgess T, Wingfield MJ, Wingfield BD (2001b) Simple sequence repeat markers distinguished among morphotypes of Sphaeropsis sapinea. Appl Environ Microbiol 67:354–362PubMedCrossRefGoogle Scholar
  6. Burgess T, Gordon TR, Wingfield MJ, Wingfield BD (2004a) Geographic isolation of Diplodia scrobiculata and its association with native Pinus radiata. Mycol Res 108:1399–1406PubMedCrossRefGoogle Scholar
  7. Burgess T, Wingfield MJ, Wingfield BD (2004b) Global distribution of Diplodia pinea genotypes revealed using simple sequence repeat (SSR) markers. Australas Plant Pathol 33:513–519CrossRefGoogle Scholar
  8. Burgess TI, Sakalidis ML, Hardy GES (2006) Gene flow of the canker pathogen Botryosphaeria australis between Eucalyptus globulus plantations and native eucalypt forests in Western Australia. Austral Ecol 31:559–566CrossRefGoogle Scholar
  9. Burgess TI, Bihon W, Wingfield MJ, Wingfield BD (2009) A simple and rapid method to determine vegetative compatibility groups in fungi. Inoculum Mycol Soc Am 60:1–2Google Scholar
  10. Dakin N, White D, Hardy G, Burgess TI (2010) The opportunistic pathogen, Neofusicoccum australe, is responsible for crown dieback of peppermint (Agonis flexuosa) in Western Australia. Australas Plant Pathol 39:1–5CrossRefGoogle Scholar
  11. Desprez-Loustau ML, Marcais B, Nageleisen LM, Piou’ D, Vannini A (2006) Interactive effects of drought and pathogens in forest trees. Ann For Sci 63:597–612CrossRefGoogle Scholar
  12. Flowers J, Nuckles E, Hartman J, Vaillancourt L (2001) Latent infections of Austrian and Scots pine tissues by Sphaeropsis sapinea. Plant Dis 85:1107–1112CrossRefGoogle Scholar
  13. Flowers J, Hartman J, Vaillancourt L (2003) Detection of latent Sphaeropsis sapinea infections in Austrian pine tissues using nested-polymerase chain reaction. Phytopathology 93:1471–1477PubMedCrossRefGoogle Scholar
  14. Gamboa MA, Bayman P (2001) Communities of Endophytic Fungi in Leaves of a Tropical Timber Tree (Guarea guidonia: Meliaceae). Biotropica 33:352–360Google Scholar
  15. Leslie JF (1993) Fungal vegetative compatibility. Annu Rev Phytopathol 31:127–150PubMedCrossRefGoogle Scholar
  16. Lundquist JE (1987) A history of five forest diseases in South Africa. S Afr For J 140:51–59Google Scholar
  17. Maresi G, Luchi N, Pinzani P, Pazzagli M, Capretti P (2007) Detection of Diplodia pinea in asymptomatic pine shoots and its relation to the Normalized Isolation index. For Pathol 37:272–280Google Scholar
  18. McCutcheon TL, Carroll GC (1993) Genotypic Diversity in Populations of a Fungal Endophyte from Douglas Fir. Mycologia 85:180–186CrossRefGoogle Scholar
  19. McDonald BA, McDermott JM (1993) Population genetics of plant pathogenic fungi: electrophoresis markers give unprecedented precision to analysis of genetic structure of populations. Bioscience 43:311–319CrossRefGoogle Scholar
  20. Milgrom MG, Cortesi P (1999) Analysis of population structure of the chestnut blight fungus based on vegetative incompatibilty genotypes. Proc Natl Acad Sci USA 96:10518–10523CrossRefGoogle Scholar
  21. Milgroom MG, Sotirovski K, Spica D, Davis JE, Brewer MT, Milev M, Cortesi P (2008) Clonal population structure of the chestnut blight fungus in expanding ranges in south Eastern Europe. Mol Ecol 17:4446–4458PubMedCrossRefGoogle Scholar
  22. Müller MM, Valjakka R, Suokko A, Hantula J (2001) Diversity of endophytic fungi of single Norway spruce needles and their role as pioneer decomposers. Mol Ecol 10:1801–1810PubMedCrossRefGoogle Scholar
  23. Nei M (1973) Analysis of gene diversity in subdivided populations. Proc Natl Acad Sci USA 70:3321–3323PubMedCrossRefGoogle Scholar
  24. Santana QC, Coetzee MPA, Steenkamp ET, Mlonyeni OX, Hammond GNA, Wingfield MJ, Wingfield BD (2009) Microsatellite discovery by deep sequencing of enriched genomic libraries. Biotechniques 46:217–223PubMedCrossRefGoogle Scholar
  25. Santini A, Pepori A, Ghelardini L, Capretti P (2008) Persistence of some pine pathogens in coarse woody debris and cones in a Pinus pinea forest. For Ecol Manage 256:502–506CrossRefGoogle Scholar
  26. Smith H, Wingfield MJ, Crous PW, Coutinho TA (1996) Sphaeropsis sapinea and Botryosphaeria dothidea endophytic in Pinus spp. and Eucalyptus spp. in South Africa. S Afr J Bot 62:86–88Google Scholar
  27. Smith H, Wingfield MJ, de Wet J, Coutinho TA (2000) Genotypic diversity of Sphaeropsis sapinea from South Africa and Northern Sumatra. Plant Dis 84:139–142CrossRefGoogle Scholar
  28. Smith H, Wingfied MJ, Couinho TA (2002) The role of latent Sphaeropsis sapinea infectins in post-hail associated die-back of Pinus paula. For Ecol Manag 164:177–184CrossRefGoogle Scholar
  29. Stanosz GR, Smith DR, Guthmiller MA, Stanosz JC (1997) Persistence of Sphaeropsis sapinea on or in asymptomatic shoots of red and jack pines. Mycologia 89:525–530CrossRefGoogle Scholar
  30. Stanosz GR, Blodgett JT, Kruger EL (2001) Water stress and Sphaeropsis sapinea as a latent pathogen of red pine seedlings. New Phytol 149:531–538CrossRefGoogle Scholar
  31. Stanosz GR, Smith DR, Albers JS (2005) Surveys for asymptomatic persistence of Sphaeropsis sapinea on or in stems of red pine seedlings from seven Great Lakes region nurseries. For Pathol 35:233–244Google Scholar
  32. Swart WJ, Wingfield MJ (1991) Biology and control of Sphaeropsis sapinea on Pinus species in South Africa. Plant Dis 75:761–766CrossRefGoogle Scholar
  33. Swart WJ, Wingfield MJ, Knox-Davies PS (1987) Factors associated with Sphaeropsis sapinea infection of pine trees in South Africa. Phytophylactica 19:505–510Google Scholar
  34. Swofford DL (2002) PAUP*. Phylogenetic Analysis Using Parsimony (* and other Methods), Version 4. Sinauer Associates, SunderlandGoogle Scholar
  35. Van der Nest MA, Steenkamp ET, Wingfield BD, Wingfield MJ (2000) Development of simple sequence repeats (SSR) markers in Eucalyptus from amplified inter-simple sequence repeats (ISSR). Plant Breed 119:433–436CrossRefGoogle Scholar
  36. Vanneste JL, Robert AH, Stuart JK, Roberta LF, Patrick TH (2002) Biological control of sapstain fungi with natural products and biological control agents: review of the work carried out in New Zealand. Mycol Res 106:228–232CrossRefGoogle Scholar
  37. Yeh FC, Yang RC, Boyle T (1999) PopGene version 1.31 (Microsoft windows based freeware for population genetic analysis. Alberta).Google Scholar
  38. Zane L, Bargelloni L, Patarnello T (2002) Strategies for microsatellite isolation: a review. Mol Ecol 11:1–16PubMedCrossRefGoogle Scholar
  39. Zwolinski JB, Swart WJ, Wingfield MJ (1990) Intensity of die-back induced by Sphaeropsis sapinea in relation to site conditions. Eur J For Pathol 20:167–174CrossRefGoogle Scholar

Copyright information

© Australasian Plant Pathology Society Inc. 2011

Authors and Affiliations

  • Wubetu Bihon
    • 1
  • Treena Burgess
    • 1
    • 2
  • Bernard Slippers
    • 1
  • Michael J. Wingfield
    • 1
  • Brenda D. Wingfield
    • 1
  1. 1.Department of Genetics, Forestry and Agricultural Biotechnology InstituteUniversity of PretoriaPretoriaSouth Africa
  2. 2.School of Biological Sciences and BiotechnologyMurdoch UniversityPerthAustralia

Personalised recommendations