Multiple gene genealogies reveal important relationships between species ofPhaeophleosporainfectingEucalyptusleaves

Abstract

The majority of Eucalyptus species are native to Australia, but worldwide there are over 3 million ha of exotic plantations, especially in the tropics and subtropics. Of the numerous known leaf diseases, three species of Phaeophleospora can cause severe defoliation of young Eucalyptus; Phaeophleospora destructans, Phaeophleospora eucalypti and Phaeophleospora epicoccoides. Phaeophleospora destructans has a major impact on seedling survival in Asia and has not, as yet, been found in Australia where it is considered a serious threat to the biosecurity of native eucalypts. It can be difficult to distinguish Phaeophleospora species based on symptoms and micromorphology and an unequivocal diagnostic tool for quarantine purposes would be useful. In this study, a multiple gene genealogy of these Phaeophleospora species and designed specific primers has been constructed to detect their presence from leaf samples. The phylogenetic position of these Phaeophleospora species within Mycosphaerella was established. They are closely related to each other and to other important Eucalyptus pathogens, Mycosphaerella nubilosa, Mycosphaerella cryptica and Colletogloeopsis zuluensis. The specific primers developed can now be used for diagnostic and screening purposes within Australia.

Fishing for Phytophthora from Western Australia’s waterways: a distribution and diversity survey

“…The PCR reaction mixture and conditions were as described by Andjic et al (2007). For identification purposes templates were sequenced with ITS-4.…”

“…Harvested mycelium was frozen in liquid nitrogen, ground to a fine powder and genomic DNA was extracted according to Andjic et al (2007).…”

Phytophthora bilorbang sp. nov., a new species associated with the decline of Rubus anglocandicans (European blackberry) in Western Australia

“…Harvested mycelium was frozen in liquid nitrogen and ground to a fine powder. Genomic DNA was extracted according to the method of Andjic et al (2007). The region spanning the internal transcribed spacer (ITS1-5.8S-ITS2) region of the ribosomal DNA was amplified using the primers DC6 (Cooke et al 2000) and ITS-4 (White et al 1990).…”

Potential for dissemination of Phytophthora cinnamomi by feral pigs via ingestion of infected plant material

“…The mycelium was harvested by scraping it from the agar surface with a sterile blade and placing it in a 1.5 ml sterile Eppendorf tube. Harvested mycelium was frozen in liquid nitrogen, ground to a fine powder and genomic DNA extracted according to Andjic et al (2007). The region spanning the internal transcribed spacer (ITS) 1-5.8S-ITS2 region of rDNA was amplified using primers ITS6 (5' GAA GGT GAA GTC GTA ACA AGG 3') (Cooke et al, 2000) and ITS4 (5' TCC TCC GCT TAT TGA TAT GC 3') (White et al, 1990).…”

“…The region spanning the internal transcribed spacer (ITS) 1-5.8S-ITS2 region of rDNA was amplified using primers ITS6 (5' GAA GGT GAA GTC GTA ACA AGG 3') (Cooke et al, 2000) and ITS4 (5' TCC TCC GCT TAT TGA TAT GC 3') (White et al, 1990). The PCR reaction mixture, amplification conditions, clean up of products and sequencing were as described by Andjic et al (2007).…”