Association of Phytophthora with Declining Vegetation in an Urban Forest Environment.

Urban forests consist of various environments from intensely managed spaces to conservation areas and are often reservoirs of a diverse range of invasive pathogens due to their introduction through the nursery trade. Pathogens are likely to persist because the urban forest contains a mixture of native and exotic plant species, and the environmental conditions are often less than ideal for the trees. To test the impact of different land management approaches on the Phytophthora community, 236 discrete soil and root samples were collected from declining trees in 91 parks and nature reserves in Joondalup, Western Australia (WA). Sampling targeted an extensive variety of declining native trees and shrubs, from families known to be susceptible to Phytophthora. A sub-sample was set aside and DNA extracted for metabarcoding using Phytophthora-specific primers; the remaining soil and root sample was baited for the isolation of Phytophthora. We considered the effect on the Phytophthora community of park class and area, soil family, and the change in canopy cover or health as determined through sequential measurements using remote sensing. Of the 236 samples, baiting techniques detected Phytophthora species from 24 samples (18 parks), while metabarcoding detected Phytophthora from 168 samples (64 parks). Overall, forty-four Phytophthora phylotypes were detected. Considering only sampling sites where Phytophthora was detected, species richness averaged 5.82 (range 1-21) for samples and 9.23 (range 2-24) for parks. Phytophthora multivora was the most frequently found species followed by P. arenaria, P. amnicola and P. cinnamomi. While park area and canopy cover had a significant effect on Phytophthora community the R2 values were very low, indicating they have had little effect in shaping the community. Phytophthora cinnamomi and P. multivora, the two most invasive species, often co-occurring (61% of samples); however, the communities with P. multivora were more common than those with P. cinnamomi, reflecting observations over the past decade of the increasing importance of P. multivora as a pathogen in the urban environment.

Seedling mycorrhizal type and soil chemistry are related to canopy condition of Eucalyptus gomphocephala.

The health of Eucalyptus gomphocephala is declining within its natural range in south-western Australia. In a pilot study to assess whether changes in mycorrhizal fungi and soil chemistry might be associated with E. gomphocephala decline, we set up a containerized bioassay experiment with E. gomphocephala as the trap plant using intact soil cores collected from 12 sites with E. gomphocephala canopy condition ranging from healthy to declining. Adjacent soil samples were collected for chemical analysis. The type of mycorrhiza (arbuscular or ectomycorrhizal) formed in containerized seedlings predicted the canopy condition of E. gomphocephala at the sites where the cores were taken. Ectomycorrhizal fungi colonization was higher in seedling roots in soil taken from sites with healthy canopies, whereas colonization by arbuscular mycorrhizal fungi dominated in roots in soil taken from sites with declining canopies. Furthermore, several soil chemical properties predicted canopy condition and the type of mycorrhizal fungi colonizing roots. These preliminary findings suggest that large-scale studies should be undertaken in the field to quantify those ectomycorrhiza (ECM) fungi sensitive to E. gomphocephala canopy decline and whether particular ECM fungi are bioindicators of ecosystem health.

Botryosphaeriaceae from tuart (Eucalyptus gomphocephala) woodland, including descriptions of four new species.

Eucalyptus gomphocephala (tuart) is a tree native to the southwest coast of Western Australia, where, in some areas, there is a significant decline in the health of tuart. Botryosphaeriaceous taxa have been isolated as endophytes and canker pathogens from numerous hosts in many parts of the world and have been implicated in the decline of E. gomphocephala. In the present study, endophytic fungi were isolated from a wide variety of native woody plant species (Acacia cochlearis, A. rostellifera, Allocasuarina fraseriana, Agonis flexuosa, Banksia grandis, E. gomphocephala, E. marginata and Santalum acuminatum), at two locations in native E. gomphocephala woodland; a site in decline at Yalgorup National Park and a healthy site at Woodman Point Regional Park. Of the 226 isolates obtained, 154 were botryosphaeriaceous taxa, 80% of which were found to be Neofusicoccum australe, isolated from all hosts at both collection sites. Four new species are described, Dothiorella moneti, Dothiorella santali, Neofusicoccum pennatisporum, and a species belonging to a genus only recently included in the Botryosphaeriaceae, Aplosporella yalgorensis. The other species isolated were Botryosphaeria dothidea on the new hosts A. rostellifera, A. cochlearis and E. marginata and Dichomera eucalypti, on the new host E. marginata. None of the new species formed lesions on excised stems of their host species, E. gomphocephala, or a common plantation species, E. globulus. However, Neofusicoccum australe formed lesions on excised stems of E. globulus and E. gomphocephala.

Phylogenetic reassessment supports accommodation of Phaeophleospora and Colletogloeopsis from eucalypts in Kirramyces.

Species of Phaeophleospora are anamorphs of Mycosphaerella and they include some of the most serious foliar pathogens of Eucalyptus spp. grown in plantations worldwide. Pathogens assigned to this genus and occurring on Eucalyptus spp. were previously treated in Kirramyces and they are also phylogenetically closely related to other anamorphs of Mycosphaerella residing in the genus Colletogloeopsis. The primary aim of this study was to consider the appropriate taxonomic placement of these species. To achieve this goal, morphological characteristics and DNA sequence data from the ITS and translation EF1-alpha gene regions were used to compare the type species P. eugeniae, Phaeophleospora spp. and Colletogloeopsis spp. occurring on eucalypts, using ex-type cultures and herbarium specimens. Phylogenetic data and morphological comparisons supported the separation of P. eugeniae from Phaeophleospora species occurring on eucalypts. The name Phaeophleospora is retained for P. eugeniae and the name Kirramyces is resurrected for the species occurring on eucalypts (genera Eucalyptus, Corymbia, and Angophora). Sequence data from the type specimens of two previously described species of Kirramyces, K. lilianiae and K. delegatensis, show they reside in a clade with other Kirramyces spp. Morphological and DNA sequence comparisons also showed that there is considerable overlap between species of Phaeophleospora and Colletogloeopsis from eucalypts. Based on these findings, Colletogloeopsis is reduced to synonymy with the older Kirramyces and the description of Kirramyces is emended to include species with aseptate, as well as multiseptate, conidia produced in acervuli or pycnidia. Two new species of Kirramyces, K. angophorae and K. corymbiae, are also described.

Three new Lasiodiplodia spp. from the tropics, recognized based on DNA sequence comparisons and morphology.

Botryosphaeria rhodina (anamorph Lasiodiplodia theobromae) is a common endophyte and opportunistic pathogen on more than 500 tree species in the tropics and subtropics. During routine disease surveys of plantations in Australia and Venezuela several isolates differing from L. theobromae were identified and subsequently characterized based upon morphology and ITS and EF1-alpha nucleotide sequences. These isolates grouped into three strongly supported clades related to but different from the known taxa, B. rhodina and L. gonubiensis, These have been described here as three new species L. venezuelensis sp. nov., L. crassispora sp. nov. and L. rubropurpurea sp. nov. The three could be distinguished easily from each other and the two described species of Lasiodiplodia, thus confirming phylogenetic separations. Furthermore all five Lasiodiplodia spp. now recognized separated from Diplodia spp. and Dothiorella spp. with 100% bootstrap support.

Botryosphaeria species from Eucalyptus in Australia are pleoanamorphic, producing dichomera synanamorphs in culture.

Species within the genus Botryosphaeria include some of the most widespread and important pathogens of woody plants, and have been the focus of numerous taxonomic studies in recent years. It is currently accepted that anamorphs of Botryosphaeria belong to two distinct genera, Fusicoccum and Diplodia. Species within the genus Fusicoccum commonly produce aseptate, hyaline conidia. In the present study, fungi were isolated from foliage and wood of Eucalyptus in native forests and plantations in Australia. Although these fungi produced Dichomera anamorphs in culture, they clustered within the Fusicoccum clade of Botryosphaeria based on their ITS sequence data. Four species, Botryosphaeria dothidea, B. parva, B. ribis and B. australis produced Dichomera conidia in culture. The Dichomera synanamorphs are described for these four species of Botryosphaeria. In addition, falling within the Fusicoccum clade of Botryosphaeria, two species were found to be distinct from previously described Botryosphaeria spp. based on their ITS sequences, but synonymous with D. versiformis and D. eucalypti. These observations are currently unique to isolates from host trees within the genus Eucalyptus in Australia, and the pleoanamorphic nature of these species is discussed.

A specific primer PCR and RFLP assay for the rapid detection and differentiation in planta of some Mycosphaerella species associated with foliar diseases of Eucalyptus globulus.

It is difficult to accurately identify Mycosphaerella species associated with leaf diseases of Eucalyptus based on morphological characters, as there is considerable overlap between very similar species and subspecies, and isolation from the host is not easy. Thus, a PCR and RFLP assay based on the ITS region of nr DNA was developed for the rapid detection and differentiation of M. nubilosa, M. cryptica and two non-sporing unidentified Mycosphaerella species isolated from the foliage of trees in resistant and susceptible families of E. globulus in a seed orchard at Kinglake West, Victoria, Australia. The M. nubilosa primer pair MNF/MNR was highly specific. A PCR-RFLP system based on the primer pair MCF/MCR, coupled with two restriction enzymes (DdeI and Tru1 I), differentiated M. cryptica, M. nubilosa, M. tasmaniensis and M. aff. vespa. One of the unidentified field-isolated Mycosphaerella species was identified as M. grandis on the basis of ITS sequence data while the other species remains unidentified. A PCR-RFLP system based on the primer pair U1F/U1R, coupled with the restriction enzyme StyI, differentiated between the two unidentified species. Unexpectedly, unlike isolation and culture studies, these assays detected M. nubilosa, M. cryptica and M. grandis in all single lesions examined on both juvenile and adult leaves, and on both highly resistant and highly susceptible E. globulus trees at this site.