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Mycosphaerella leaf disease on eucalypts in Western Australia - The diversity and impact

Jackson, Sarah (2013) Mycosphaerella leaf disease on eucalypts in Western Australia - The diversity and impact. PhD thesis, Murdoch University.

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Eucalyptus plantation forestry in Western Australia (WA) is a relatively young industry and by the end of 2008, the total plantation estate (softwood and hardwood) was over 950 000 ha. The predominant plantation species is Eucalyptus globulus, native to south-eastern Australia. In Western Australia (WA), the most serious foliar disease of eucalypt plantations is Mycosphaerella Leaf Disease (MLD). However, little systematic sampling for MLD has been carried out in WA to determine its impact on plantations, yields, species involved or whether they are introduced or not. The overall aim of this thesis was to investigate MLD in south-western Australia with a particular focus on the species diversity, taxonomy and the impact on early growth on E. globulus.

The increase in the number of Mycosphaerella and Teratosphaeria species associated with Mycosphaerella leaf disease (MLD) in E. globulus plantations in WA in the past decade has raised concern about the possible movement of pathogens between the native forests and plantations and vice versa. A survey of necrotic leaf spots collected from plantation and endemic eucalypts from WA and Queensland was conducted. Overall, ten new Eucalyptus host records for Mycosphaerella/ Teratosphaeria species were isolated from WA and five from Queensland. Significantly, M. nubilosa was isolated from E. grandis x resinifera and E. urophylla x globulus in WA. This is the first time M. nubilosa has been isolated from Eucalyptus hosts within the series Resinifera (see Chapter 2).

An assessment of the number of fungi that may be contributing to MLD in E. globulus plantations in WA was undertaken (Chapter 3) and the changes in the number of species and their incidence since the first surveys were conducted. Four new records of Mycosphaerella were identified in this study; M. ellipsoidea, P. fori, M. suttoniae and M. tasmaniensis. Mycosphaerella ellipsoidea and P. fori are first records for Australia, and M. suttoniae and M. tasmaniensis are first records for WA. The current work shows an increase in the number of Mycosphaerella species associated with plantation eucalypts in WA and Australia. With the exception of M. cryptica, none of these species were known in WA prior to the commencement of large-scale E. globulus plantations, and with M. cryptica as the exception, none have a known impact on the major native eucalypts in the region.

The ITS region of the type material of T. parva, M. grandis and M. gregaria using culture and herbarium specimens was sequenced and compared to existing sequences from GenBank (Chapter 4). This was the first study to examine and sequence the type material of M. grandis, T. parva and M. gregaria. As the sequences of the ITS region of M. grandis and T. parva were identical it was concluded that M. grandis be reduced to synonymy with T. parva. Mycosphaerella aurantia, M. buckinghamiae and M. africana also match the type sequence of M. gregaria. Therefore, these should all be synonymised to M. gregaria. Also, this study was the first to describe ITS sequence variation within the same Mycosphaerella isolate.

The aim of Chapter 5 was to identify the infection pathway at the leaf surface using scanning electron microscopy and to determine the pathogenicity of M. marksii on E. globulus. The use of glycerol as a surfactant and its effect on ascospore viability was also assessed. However, this study was unable to confirm pathogenicity of M. marksii on E. globulus seedlings under laboratory conditions. However, M. marksii ascospores were able to germinate and enter E. globulus stoma 3–6 days after initial infection.

Species-specific primers were successfully designed and tested for three Mycosphaerella species that occur on E. globulus in WA (Chapter 6). Meteorological conditions appeared to determine the defoliation of juvenile foliage and not MLD as levels of MLD remained relatively low throughout the trial period. The MLD levels increased throughout spring as warm wet conditions favoured the development of disease especially on the flush of new juvenile foliage. Also, new foliage emerged after late summer rainfall. As disease pressure mounted, the trees responded through defoliation. As temperatures increased and the juvenile foliage aged, there is likely to have been an increase in the defoliation of leaves. Therefore, by mid-summer defoliation levels reached a similar level to disease and insect damage. Following leaf defoliation and the emergence of new juvenile and adult leaves, the relative amount of disease on the trees decreased. This is because most of the disease was present on the older juvenile foliage which was shed. Field observations can be a reliable indication of disease progression. Although field observations at a branch level over exaggerated levels of MLD when there was a higher level of foliage, there was still a similar trend in the amount of disease when compared to the ASSESS program. Some experience in disease monitoring would indicate a more accurate assessment of MLD. It is interesting to note that the assessors tended to overestimate disease when MLD was at a higher level, and this also included the author.

Infection studies of Uwebraunia dekkeri were conducted to confirm how this species enters E. globulus leaves and to determine its pathogenicity (Chapter 7). This study demonstrated that conidia of U. dekkeri could infect E. globulus leaves and that it is not a hyperparasite of M. cryptica or M. nubilosa. Conidiogenesis was both percurrent and sympodial and the phenomenon of anastomosis was observed for the first time on the leaf surface.

The impact that MLD has on the wood volume has previously not been investigated in WA (Chapter 8). Through the application of pesticides and fungicides in the early stages of establishment at two plantations near Albany, tree volumes were significantly increased. However, the increase in wood volume would be offset by the pesticide and application costs. This study demonstrated that monitoring for pests and disease would be more effective than spraying of chemical treatments for the first three years. The regular use of chemical treatments is expensive to maintain and is proving to be environmentally unacceptable by some communities. This study also showed that spraying for low levels of MLD had little effect on disease incidence and/ or volume increase in E. globulus plantations in WA. The most important factors for a healthy plantation appear to be site selection, preparation and tree genetics.

This study was the first to investigate the impact of MLD on the growth of Eucalyptus globulus plantations in WA. As part of this study, the biology, taxonomy and pathogenicity of the main species present in WA were investigated. The key findings were: i) the number, abundance and distribution of Mycosphaerella/ Teratosphaeria species in WA is not static and plantations should be continually monitored for the presence of new potentially threatening species; ii) spraying for MLD, although effective in reducing the prevalence and impact on growth, was not economically viable; and iii) intragenomic variation of the ribosomal genome may explain sequence variation observed in single spore isolates of Mycosphaerella/ Teratosphaeria and this has taxonomic implications. Further work would identify the impact the new records are having on the plantation estate and also if these species have the potential to spread into the neighbouring endemic forests. This study has provided a broader understanding of MLD in WA and the development of tools that could be used for further study.

Publication Type: Thesis (PhD)
Murdoch Affiliation: School of Veterinary and Life Sciences
Supervisor: Hardy, Giles, Dell, Bernard and Maxwell, Aaron
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