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ITS-RFLP and sequence analysis of endophytes from Acianthus, Caladenia and Pterostylis (Orchidaceae) in southeastern Queensland

Published online by Cambridge University Press:  30 March 2005

Jeremy J. BOUGOURE ,
Damian S. BOUGOURE ,
John W. G. CAIRNEY  and
John D. W. DEARNALEY
Jeremy J. BOUGOURE
Affiliation:
School of Plant Biology, Faculty of Natural and Agricultural Sciences, The University of Western Australia, Crawley 6009, Australia.
Damian S. BOUGOURE
Affiliation:
Mycorrhiza Research Group, Centre for Horticulture and Plant Sciences, The University of Western Sydney, Parramatta Campus, Locked Bag 1797, Penrith South DC, NSW 1797, Australia.
John W. G. CAIRNEY
Affiliation:
Mycorrhiza Research Group, Centre for Horticulture and Plant Sciences, The University of Western Sydney, Parramatta Campus, Locked Bag 1797, Penrith South DC, NSW 1797, Australia.
John D. W. DEARNALEY
Affiliation:
Department of Biological and Physical Sciences, The University of Southern Queensland, Toowoomba 4350, Australia. E-mail: [email protected].
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Abstract

We used ITS-RFLP and sequence analysis to determine the identities of the fungal endophytes of six terrestrial orchid species from southeastern Queensland, a region previously unexplored in this context. Pure cultures of orchid – colonising fungi were obtained and fungal identities were assessed by means of ITS-PCR, RFLP analysis, sequence comparison, and protocorm colonisation tests. ITS-PCR and RFLP analysis resulted in five main groupings. Sequencing and GenBank comparison of these five groups showed that the fungal endophytes isolated from the three Pterostylis species were probably Thanatephorus species. There was close sequence identity (90%) of the fungus isolated from Acianthus spp. to Epulorhiza repens, suggesting these may be the same fungal species. However, that only E. repens succeeded in colonising protocorms of Thelymitra pauciflora suggests these may be different species of Epulorhiza. Analysis of the ITS and LSU sequences of the fungus isolated from Caladenia carnea showed high identities with a sequence from a Sebacina vermifera originally isolated from Caladenia dilatata. These results show that there is specificity for fungal partners within the orchid genera Acianthus, Caladenia and Pterostylis.

Type
Research Article
Information
Mycological Research , Volume 109 , Issue 4 , April 2005 , pp. 452 - 460
Copyright
© The British Mycological Society 2005

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