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New Disease Reports article 18 Jun 2013

Grijalba PE, Palmucci HE, Guillin E, Herrera C, 2013. Phytophthora multivora causing leaf spot on rhododendrons in Argentina. New Disease Reports 27, 20. DOI: 10.5197/j.2044-0588.2013.027.020

Rhododendron is a genus belonging to the family Ericaceae and contains over 1000 species commonly known as rhododendrons and azaleas (Dimitri, 1978). Phytophthora multivora is a species that was described in Western Australia in 2008 but before that it was misidentified as . citricola (Scott et al., 2009). During the early spring of 2011, leaf spot symptoms were observed on rhododendrons in two gardens in Tigre (northern Buenos Aires province) and in containers in a nursery near Buenos Aires city. Leaf spots were dark brown to almost black, visible on both sides near the leaf tips and margins, while tissue death continued down the leaf along the midrib (Fig. 1A). The veins under the diseased area of the leaves presented a reddish tint (Fig. 1B). Some leaves became brown and died, while in others the infection remained as spots on leaves.


Early view Forest Pathology article, 15 June 2013

Mulholland, V., Schlenzig, A., MacAskill, G. A., Green, S. (2013), Development of a quantitative real-time PCR assay for the detection of Phytophthora austrocedrae, an emerging pathogen in Britain. Forest Pathology. doi: 10.1111/efp.12058

A TaqMan real-time PCR assay was developed for Phytophthora austrocedrae, an emerging pathogen causing severe damage to juniper in Britain. The primers amplified DNA of the target pathogen down to 1 pg of extracted DNA, in both the presence and absence of host DNA, but did not amplify any of the non-target Phytophthora and fungal species tested. The assay provides a useful tool for screening juniper populations for the disease.


Early view Forest Pathology article, 10 April 2013

Than, D. J., Hughes, K. J. D., Boonhan, N., Tomlinson, J. A., Woodhall, J. W., Bellgard, S. E. (2013), A TaqMan real-time PCR assay for the detection of Phytophthora ‘taxon Agathis’ in soil, pathogen of Kauri in New Zealand. Forest Pathology. doi: 10.1111/efp.12034

Kauri Agathis australis, an iconic tree of New Zealand, is under threat from an introduced disease-causing pathogen provisionally named Phytophthora ‘taxon Agathis’ (referred to as PTA). This soilborne, Pythiaceous species belongs to the Chromista and causes a collar rot resulting in yellowing of the foliage and thinning of the canopy, which eventually causes death of the infected tree. The management and containment of this pathogen requires rapid and reliable detection in the soil. The current method for soil detection utilizes a soil bioassay involving lupin baits and soil flooding in a process that takes between ten and twenty days. We describe a real-time PCR assay based on TaqMan chemistry for the specific detection of PTA, which targets the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA. This TaqMan real-time PCR assay could be used with DNA extracted directly from bulk soil samples to enable rapid quantification of PTA within soil. The detection limit was 2 fg of PTA DNA from pure culture, or 20 fg in the presence of DNA extracted from soil. The assay was validated using soil samples taken from a PTA-infested site and soil spiked with a known concentration of oospores. We conclude that the TaqMan real-time PCR assay offers a more time-efficient method for detection of PTA in soil than existing methods.


March 2013 Biological Invasions article

Challenges in predicting invasive reservoir hosts of emerging pathogens: mapping Rhododendron ponticum as a foliar host for Phytophthora ramorum and Phytophthora kernoviae in the UK. Bethan V. Purse, Philipp Graeser, Kate Searle, Colin Edwards, Catriona Harris. Biological Invasions March 2013, Volume 15, Issue 3, pp 529-545. DOI: 10.1007/s10530-012-0305-y

Abstract: Invasive species can increase the susceptibility of ecosystems to disease by acting as reservoir hosts for pathogens. Invasive hosts are often sparsely recorded and not in equilibrium, so predicting their spatial distributions and overlap with other hosts is problematic. We applied newly developed methods for modelling the distribution of invasive species to the invasive shrub Rhododendron ponticum—a foliar reservoir host for the Phytophthora oomycete plant pathogens, P. ramorum and P. kernoviae, that threaten woodland and heathland habitat in Scotland. We compiled eleven datasets of biological records for R. ponticum (1,691 points, 8,455 polygons) and developed Maximum Entropy (MaxEnt) models incorporating landscape, soil and climate predictors. Our models produced accurate predictions of current suitable R. ponticum habitat (training AUC = 0.838; test AUC = 0.838) that corresponded well with population performance (areal cover). Continuous broad-leaved woodland cover, low elevation (<400 m a.s.l.) and intermediate levels of soil moisture (or Enhanced Vegetation Index) favoured presence of R. ponticum. The high coincidence of suitable habitat with both core native woodlands (54 % of woodlands) and plantations of another sporulation host, Larix kaempferi (64 % of plantations) suggests a high potential for spread of Phytophthora infection to woodland mediated by R. ponticum. Incorporating non-equilibrium modelling methods did not improve habitat suitability predictions of this invasive host, possibly because, as a long-standing invader, R. ponticum has filled more of its available habitat at this national scale than previously suspected.


Journal of Phytopatholgy early view article 3 April 2013

Tooley, P. W., Browning, M. and Leighty, R. M. (2013), Inoculum density relationships for infection of some eastern US forest species by Phytophthora ramorum. Journal of Phytopathology. doi: 10.1111/jph.12107

Our objectives were to establish inoculum density relationships between P. ramorum and selected hosts using detached leaf and whole-plant inoculations. Young plants and detached leaves of Quercus prinus (Chestnut oak), Q. rubra (Northern red oak), Acer rubrum (red maple), Kalmia latifolia (mountain laurel) and Rhododendron ‘Cunningham's White’ were dip-inoculated with varying numbers of P. ramorum sporangia, and the total number of diseased and healthy leaves recorded following incubation at 20°C and 100% relative humidity. Calibration threshold estimates for obtaining 50% infected leaves based on linear analysis ranged from 36 to 750 sporangia/ml for the five hosts. Half-life (LD50) estimates (the number of spores for which the per cent of diseased leaves reaches 50% of its total) from asymptotic regression analysis ranged from 94 to 319 sporangia/ml. Statistically significant differences (P = 0.0076) were observed among hosts in per cent infection in response to increased inoculum density. Inoculum threshold estimates based on studies with detached leaves were comparable to those obtained using whole plants. The results provide estimates of inoculum levels necessary to cause disease on these five P. ramorum hosts and will be useful in disease prediction and for development of pest risk assessments.



Early view Forest Pathology article, 19 March 2013

Scanu, B., Linaldeddu, B. T., Franceschini, A., Anselmi, N., Vannini, A., Vettraino, A. M. (2013), Occurrence of Phytophthora cinnamomi in cork oak forests in Italy. Forest Pathology. doi: 10.1111/efp.12039


An increasing decline and mortality of cork oak trees have been recently observed in central Italy and Sardinia Island. Following surveys conducted in three declining cork oak forests, a Phytophthora species was consistently isolated from soil samples collected from trees displaying different level of decline. Based on morphological features, growth rates at different temperatures and analysis of DNA sequences of the ITS region, all isolates were identified as Phytophthora cinnamomi Rands. This pathogen caused large brownish lesions on inoculated freshly cut branches of cork oak. It was re-isolated from all infected tissues. These findings represent the first report of P. cinnamomi on cork oak trees in Italy.


Early view Forest Pathology article, 11 March 2013

Pérez-Sierra, A., López-García, C., León, M., García-Jiménez, J., Abad-Campos, P., Jung, T. (2013), Previously unrecorded low-temperature Phytophthora species associated with Quercus decline in a Mediterranean forest in eastern Spain. Forest Pathology. doi: 10.1111/efp.12037

Oak decline has been a serious problem in Europe since the beginning of the twentieth century. In south-west Spain, Quercus ilex and Q. suber are the main affected species, and their decline has been associated with Phytophthora cinnamomi. During the last 10 years, a severe decline of Q. ilex and Q. faginea accompanied by a significant decrease in the production of acorns affecting natural regeneration was observed in the eastern part of the Iberian Peninsula. Therefore, the aim of this study was to investigate the possible involvement of Phytophthora spp. in the decline. A forest in the Natural Park ‘Carrascar de la Font Roja’ in Comunidad Valenciana (eastern Spain), which is dominated by Q. ilex and Q. faginea, was surveyed during 2010–2011. Symptomatic trees showed thinning and dieback of the crown, withering of leaves and death. An extensive loss of both lateral small woody roots and fine roots and callusing or open cankers on suberized roots were observed. Soil samples containing fine roots were baited using both Q. robur leaves and apple fruits. Six Phytophthora species were isolated: P. cryptogea, P. gonapodyides, P. megasperma, P. quercina, P. psychrophila and P. syringae. These are the first records of P. quercina and P. psychrophila on Q. faginea, of P. quercina in Spain and of P. psychrophila in mainland Spain. A soil infestation trial was conducted for 6 months under controlled conditions with 1-year-old seedlings of Q. ilex and Q. faginea. Phytophthora cinnamomi was included in the pathogenicity test for comparison. The results showed that Q. ilex seedlings were generally more susceptible to infection than Q. faginea with P. cinnamomi being the most aggressive pathogen to both oak species. The two most commonly isolated Phytophthora species, P. quercina and P. psychrophila, also proved their pathogenicity towards both Q. ilex and Q. faginea.


Early view Forest Pathology article, 13 February 2013

Crone, M., McComb, J. A., O'Brien, P. A., Hardy, G. E. S. J.Assessment of Australian native annual/herbaceous perennial plant species as asymptomatic or symptomatic hosts of Phytophthora cinnamomi under controlled conditions.  Forest Pathology. doi: 10.1111/efp.12027

Phytophthora cinnamomi is a necrotrophic pathogen of woody perennials and devastates many biomes worldwide. A controlled perlite–hydroponic system with no other hyphae-producing organisms as contaminants present allowed rapid assessment of ten annual and herbaceous perennial plant species most of which have a wide distribution within the jarrah (Eucalyptus marginata) forest in Western Australia where this pathogen has been introduced. As some annuals and herbaceous perennials have recently been reported as symptomatic and asymptomatic hosts, laboratory screening of some of the field-tested annuals and herbaceous perennials and additional species was used to further evaluate their role in the pathogen's disease cycle. Nine of the species challenged with the pathogen were asymptomatic, with none developing root lesions; however, Trachymene pilosa died. The pathogen produced thick-walled chlamydospores and stromata in the asymptomatic roots. Furthermore, haustoria were observed in the roots, indicating that the pathogen was growing as a biotroph in these hosts.


Early view Forest Pathology article, 31 January 2013

Jung, T., Colquhoun, I. J., Hardy, G. E. St. J. (2013), New insights into the survival strategy of the invasive soilborne pathogen Phytophthora cinnamomi in different natural ecosystems in Western Australia. Forest Pathology. doi: 10.1111/efp.12025

Despite its importance as one of the most notorious, globally distributed, multihost plant pathogens, knowledge on the survival strategy of Phytophthora cinnamomi in seasonally dry climates is limited. Soil and fine roots were collected from the rhizosphere of severely declining or recently dead specimens of 13 woody species at 11 dieback sites and two dieback spots and from healthy specimens of five woody species at four dieback-free sites in native forests, woodlands and heathlands of the south-west of Western Australia (WA). Phytophthora cinnamomi was recovered from 80.4, 78.1 and 100% of tested soil, fine root and soil–debris slurry samples at the 11 dieback sites, in some cases even after 18-month storage under air-dry conditions, but not from the small dieback spots and the healthy sites. Direct isolations from soil–debris slurry showed that P. cinnamomi colonies exclusively originated from fine roots and root fragments not from free propagules in the soil. Microscopic investigation of P. cinnamomi-infected fine and small woody roots and root fragments demonstrated in 68.8, 81.3 and 93.8% of samples from nine woody species the presence of thick-walled oospores, stromata-like hyphal aggregations and intracellular hyphae encased by lignitubers, respectively, while thin-walled putative chlamydospores were found in only 21.2% of samples from five woody species. These findings were confirmed by microscopic examination of fine roots from artificially inoculated young trees of 10 woody species. It is suggested that (i) the main function of chlamydospores is the survival in moderately dry conditions between consecutive rain events and (ii) selfed oospores, hyphal aggregations, and encased hyphae and vesicles in infected root tissue of both host and non-host species are the major long-term survival propagules of P. cinnamomi during the extremely dry summer conditions in WA.


Early view Plant Pathology article

Prospero, S., Vercauteren, A., Heungens, K., Belbahri, L. and Rigling, D. (2013). Phytophthora diversity and the population structure of Phytophthora ramorum in Swiss ornamental nurseries. Plant Pathology. doi: 10.1111/ppa.12027

Invasive oomycete pathogens have been causing significant damage to native ecosystems worldwide for over a century. A recent well-known example is Phytophthora ramorum, the causal agent of sudden oak death, which emerged in the 1990s in Europe and North America. In Europe, this pathogen is mainly restricted to woody ornamentals in nurseries and public greens, while severe outbreaks in the wild have only been reported in the UK. This study presents the results of the P. ramorum survey conducted in Swiss nurseries between 2003 and 2011. In all 120 nurseries subjected to the plant passport system, the main P. ramorum hosts were visually checked for above ground infections. Phytophthora species were isolated from tissue showing symptoms and identified on the basis of the morphological features of the cultures and sequencing of the ribosomal ITS region. Phytophthora was detected on 125 plants (66 Viburnum, 58 Rhododendron and one Pieris). Phytophthora ramorum was the most frequent species (59·2% of the plants), followed by P. plurivora, P. cactorum, P. citrophthora, P. cinnamomi, P. cactorum/P. hedraiandra, P. multivora and P. taxon PgChlamydo. The highest incidence of P. ramorum was observed on Viburnum × bodnantense. Microsatellite genotyping showed that the Swiss P. ramorum population is highly clonal and consists of seven genotypes (five previously reported in Europe, two new), all belonging to the European EU1 clonal lineage. It can therefore be assumed that P. ramorum entered Switzerland through nursery trade. Despite sanitation measures, repeated P. ramorum infections have been recorded in seven nurseries, suggesting either reintroduction or unsuccessful eradication efforts.