N. Williams

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

During one spring season, 12 Phytophthora species, two Phytophthora hybrids, three Halophytophthora species and three Phytopythium species, were isolated from 48 waterways across Western Australia. The waterways were sampled using nylon mesh bags containing leaf baits of up to six different plant species and were isolated by plating necrotic lesions on these onto Phytophthora-selective agar media. Phytophthora species were isolated from all except one waterway. Of the Phytophthora species isolated, eight are known while the remaining four are undescribed taxa. Six of the Phytophthora species and the two hybrids are from clade 6. The two hybrids and P. inundata were the predominant species recovered. Recoveries from different plant leaf baits varied with the best two baits being Pittosporum undulatum and Banksia attentuata; and from these two combined all Phytophthora species were isolated. There was a marked difference in the Phytophthora diversity in the waterways from different regions. This is the first comprehensive study from Australia to examine the Phytophthora communities in waterways, and advances our understanding of the role of these oomycetes in natural and anthropized ecosystems.

Defining the phosphite-regulated transcriptome of the plant pathogen Phytophthora cinnamomi

Phosphite, an analog of phosphate is used to control oomycete diseases on a wide range of horticultural crops and in native ecosystems. In this study, we investigated morphological and transcriptional changes induced in Phytophthora cinnamomi by phosphite. Cytological observations revealed that phosphite caused hyphal distortions and lysis of cell walls and had an adverse effect on hyphal growth. At the molecular level, the expression levels of 43 transcripts were changed. Many of these encoded proteins involved in cell wall synthesis, or cytoskeleton functioning. The results of both the microscopic and molecular investigations are consistent with phosphite inhibiting the function of the cytoskeleton and cell wall synthesis.

Pathogenicity of Phytophthora pluvialis to Pinus radiata and its relation with red needle cast disease in New Zealand

BackgroundRed needle cast, a new foliage disease of Pinus radiata in New Zealand is described. The disease has been variable in incidence and severity both regionally and in different years. The early symptoms of discrete olive coloured lesions, often with a narrow dark resinous mark or band, were first recognised in winter of 2008 in plantation forests on the eastern coast of the North Island. These lesions develop further to result in rapid needle senescence and premature defoliation. The disease has been termed red needle cast in New Zealand as affected trees have a reddish appearance prior to the casting of the needles. The subsequent four years of monitoring have confirmed that, depending on location, symptoms are first observed in late autumn through late winter. Newly developing spring and summer foliage is seldom affected. Isolation from needles using a Phytophthora-selective medium frequently yielded an unknown species of Phytophthora which was subsequently found to be identical to Phytophthora pluvialis, a species described from Oregon, USA in 2013 where it is not associated with disease. Infection appears to be limited to the needles with no recoveries of Phytophthora pluvialis having been made from the roots, stems or branches. Occasionally a second species of Phytophthora, P. kernoviae, was also recovered from needles with the same symptoms.MethodsNeedle symptoms were described in the field from 2008-2012 with isolation onto Phytophthora selective media. Koch’s postulates was completed on potted plants and detached needles.ResultsSymptoms were reproduced on both detached needles and potted plants of Pinus radiata when inoculated with z oospore suspensions of Phytophthora pluvialis.ConclusionsThis paper presents evidence that Phytophthora pluvialis is the primary cause of red needle cast in New Zealand.

Genome sequences of six Phytophthora species associated with forests in New Zealand.

In New Zealand there has been a long association of Phytophthora diseases in forests, nurseries, remnant plantings and horticultural crops. However, new Phytophthora diseases of trees have recently emerged. Genome sequencing has been performed for 12 Phytophthora isolates, from six species: Phytophthora pluvialis, Phytophthora kernoviae, Phytophthora cinnamomi, Phytophthora agathidicida, Phytophthora multivora and Phytophthora taxon Totara. These sequences will enable comparative analyses to identify potential virulence strategies and ultimately facilitate better control strategies. This Whole Genome Shotgun data have been deposited in DDBJ/ENA/GenBank under the accession numbers LGTT00000000, LGTU00000000, JPWV00000000, JPWU00000000, LGSK00000000, LGSJ00000000, LGTR00000000, LGTS00000000, LGSM00000000, LGSL00000000, LGSO00000000, and LGSN00000000.

Foliar phosphite application has minor phytotoxic impacts across a diverse range of conifers and woody angiosperms.

Phytophthora plant pathogens cause tremendous damage in planted and natural systems worldwide. Phosphite is one of the only effective chemicals to control broad-scale Phytophthora disease. Little work has been done on the phytotoxic effects of phosphite application on plant communities especially in combination with plant physiological impacts. Here, we tested the phytotoxic impact of phosphite applied as foliar spray at 0, 12, 24 and 48 kg a.i. ha(-1) . Eighteen-month-old saplings of 13 conifer and angiosperm species native to New Zealand, and two exotic coniferous species were treated and the development of necrotic tissue and chlorophyll-a-fluorescence parameters (optimal quantum yield, Fv /Fm ; effective quantum yield of photosystem II, ΦPSII ) were assessed. In addition, stomatal conductance (gs ) was measured on a subset of six species. Significant necrosis assessed by digital image analysis occurred in only three species: in the lauraceous canopy tree Beilschmiedia tawa (8-14%) and the understory shrub Dodonaea viscosa (5-7%) across phosphite concentrations and solely at the highest concentration in the myrtaceous pioneer shrub Leptospermum scoparium (66%). In non-necrotic tissue, Fv /Fm , ΦPSII and gs remained unaffected by the phosphite treatment. Overall, our findings suggest minor phytotoxic effects resulting from foliar phosphite application across diverse taxa and regardless of concentration. This study supports the large-scale use of phosphite as a management tool to control plant diseases caused by Phytophthora pathogens in plantations and natural ecosystems. Long-term studies are required to ascertain potential ecological impacts of repeated phosphite applications.

First Report of Phytophthora pluvialis Causing Needle Loss and Shoot Dieback on Douglas-fir in Oregon and New Zealand

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Development of a high throughput optical density assay to determine fungicide sensitivity of oomycetes

A high-throughput assay was developed to screen Phytophthora species for fungicide sensitivity using optical density measurements for unbiased, automated measurement of mycelial growth. The efficacy of the optical density assay (OD) to measure phosphite sensitivity in Phytophthora species was compared to two widely used methods, radial growth (RG) and dry weight (DW) assays. Three isolates of each of Phytophthora cinnamomi, P. multivora and P. pluvialis, with known phosphite exposure and three isolates of each species with no prior phosphite exposure, were screened for phosphite sensitivity using the three assays. Mycelial growth measurements were taken after culturing for 6, 14 and 15 days for the OD, DW and RG assays respectively. Mycelial growth inhibition at 15, 80, 200 and 500 μg/mL phosphite relative to growth on control media was used to determine effective concentration values for 50% growth reduction (EC50). The species varied in their tolerance to phosphite with P. cinnamomi being the least sensitive followed by P. multivora and P. pluvialis. No significant differences in tolerance were found between isolates within the same species using any method. The OD assay produced comparable EC50 values to the RG and DW assays. The growth of the three species was more sensitive to phosphite in the DW than the RG and OD assays, however limited sample throughput and greater variation in measuring small amounts of mycelia in the dry weight assessment increase variability and limits throughput. The OD assay offers a fast method to enable an inventory of chemical resistance and is particularly advantageous for slow growing species as it requires less time and offers greater throughput than existing RG and DW methods.