Cinnamomi Infection Research Articles

Spatio-temporal analysis of oak decline process in open woodlands: A case study in SW Spain

This study aims to characterize at landscape level the spatio-temporal dynamics of a massive oak decline that is occurring in dehesas ecosystems. We are looking at possibilities of matching with Phytophthora disease behavior, a harmful disease detected in the studied area, in order to interpret its implications within the context of the disease management. Spatial locations of affected trees from 2001, 2009 and 2016 identified through photointerpretation were analyzed with the inhomogeneous Ripley's K-function to assess their spatial pattern. Multivariate Adaptive Regression Splines (MARS), a non-parametric data mining method, was used to investigate the influence of a range of landscape descriptors of different nature on the proneness to oak decline, using the location of affected trees in comparison with that of healthy spots (points randomly extracted from areas covered by healthy trees).Affected trees showed a strong clustering pattern that decreased over time. The reported spatial patterns align with the hypothesis of Phytophthora cinnamomi Rands. being the main cause of oak decline in Mediterranean forests. Location of affected trees detected in different years was found to be spatially related, suggesting the implication of a contagion process. MARS models from 2001, 2009 and 2016 reported Area Under the Curve (AUC) values of 0.707, 0.671 and 0.651, respectively. Slope was the most influential landscape descriptor across the three years, with distance to afforestations being the second for 2001 and 2009. Landscape descriptors linked to human factors and soil water content seem to influence oak decline caused by Phytophthora cinnamomi at landscape level. Afforestations carried out as part of the afforestation subsidy program promoted by the European Commission in 1992 could have acted as an initial source of Phytophthora cinnamomi infection. These findings together with the consideration of the spatial and temporal scale of the spreading are essential when planning the management of oak decline in open woodlands.

Detecting symptoms of Phytophthora cinnamomi infection in Australian native vegetation using reflectance spectrometry: complex effects of water stress and species susceptibility

Diseases in natural and agricultural systems have been linked to species of the Oomycete genus Phytophthora, around the world. Direct detection of the pathogen requires sampling of soil or plant material, which can be expensive, difficult to obtain and error-prone. As an alternative, reflectance spectroscopy provides a potential indirect method for detecting symptoms of infection by P. cinnamomi. Here we evaluate the use of reflectance spectroscopy to detect physiological changes associated with infection in host plants using spectral indices designed to quantify changes in plant pigments (pigment indices), leaf water content (water indices) and fluorescence (fluorescence indices). Two grasses and two tree species with different susceptibilities to P. cinnamomi were inoculated and/or exposed to water stress in a glasshouse experiment. Inoculated plants were detected using pigment and fluorescence indices, which also had the capacity to separate inoculated plants from water stressed uninoculated plants. While inoculation may have caused an opposing spectral response to water stress in some indices, plants that were both water stressed and inoculated then demonstrated an intermediate response. Water stress was detected using the water indices in all four species, and spectroscopic changes associated with inoculation were often greater in the susceptible species. Our results indicate that reflectance spectroscopy at the leaf scale detects the effects of P. cinnamomi infection in native vegetation. Extending these results has the potential to improve early detection of disease in natural vegetation and avoiding manual sampling, thus improving management of the disease.

Exploring interactive effects of climate change and exotic pathogens on Quercus suber performance: Damage caused by Phytophthora cinnamomi varies across contrasting scenarios of soil moisture

Climate change and exotic pests and pathogens are causing alarming forest declines worldwide. However, we still lack a comprehensive understanding of how damage caused by exotic pests and pathogens might vary under the different scenarios of water availability imposed by a changing climate, particularly in water-limited forests as those that occupy Mediterranean areas. In this paper we aimed to experimentally analyse the interactive effects of the aggressive exotic pathogen Phytophthora cinnamomi and climate change-related reductions in soil moisture on seedling performance of the Mediterranean host Quercus suber. We conducted a full-factorial greenhouse experiment where the physiology and growth of Q. suber seedlings was measured in soils with different combinations of P. cinnamomi inoculum density (0, 30, 60 and 120 colony forming units per gram of dry soil) and soil moisture (15%, 40%, 50% and 100% soil water holding capacity) simulating different invasion and climate change scenarios. We found additive effects of P. cinnamomi and drought on Q. suber performance aboveground, although these effects were not always negative. In fact, seedlings showed a compensatory physiological response to P. cinnamomi infection by increasing their net photosynthetic rates. Our results also supported important interactive effects of pathogens and soil moisture on belowground performance. Thus, the inoculum density in the soil required to cause significant root damage in experimental seedlings decreased as soil moisture increased. From a climate change perspective, these results suggest that an average drier climate might imply sub-optimal conditions for P. cinnamomi infections allowing for a slower advance of the disease in invaded areas. However, this effect will be modulated by the also predicted more frequent extreme climatic events. A higher frequency of extreme rain events that saturate the soil might be particularly beneficial for P. cinnamomi, boosting its soil density beyond any possible response capacity of susceptible hosts.

Resistance to Phytophthora cinnamomi in American Chestnut (Castanea dentata) Backcross Populations that Descended from Two Chinese Chestnut (Castanea mollissima) Sources of Resistance.

Restoration of American chestnut (Castanea dentata) depends on combining resistance to both the chestnut blight fungus (Cryphonectria parasitica) and Phytophthora cinnamomi, which causes Phytophthora root rot, in a diverse population of C. dentata. Over a 14-year period (2004 to 2017), survival and root health of American chestnut backcross seedlings after inoculation with P. cinnamomi were compared among 28 BC3, 66 BC4, and 389 BC3F3 families that descended from two BC1 trees (Clapper and Graves) with different Chinese chestnut grandparents. The 5% most resistant Graves BC3F3 families survived P. cinnamomi infection at rates of 75 to 100% but had mean root health scores that were intermediate between resistant Chinese chestnut and susceptible American chestnut families. Within Graves BC3F3 families, seedling survival was greater than survival of Graves BC3 and BC4 families and was not genetically correlated with chestnut blight canker severity. Only low to intermediate resistance to P. cinnamomi was detected among backcross descendants from the Clapper tree. Results suggest that major-effect resistance alleles were inherited by descendants from the Graves tree, that intercrossing backcross trees enhances progeny resistance to P. cinnamomi, and that alleles for resistance to P. cinnamomi and C. parasitica are not linked. To combine resistance to both C. parasitica and P. cinnamomi, a diverse Graves backcross population will be screened for resistance to P. cinnamomi, survivors bred with trees selected for resistance to C. parasitica, and progeny selected for resistance to both pathogens will be intercrossed.

Regulation by biotic stress of tannins biosynthesis in Quercus ilex: Crosstalk between defoliation and Phytophthora cinnamomi infection.

Sustainability of the Mediterranean forest is threatened by oak decline, a disease of holm oak and other Quercus species that is initiated by infection with the oomycete Phytophthora cinnamomi. Focusing on the role of tannins in the chemical defense of plants, this work investigated whether tannins content in Quercus ilex is regulated by biotic stress. Screening of published genomes allowed the identification of Quercus sequences encoding enzymes for early steps of the biosynthesis of phenolic compounds, like hydrolysable tannins and condensed tannins (CT) among others, plus genes involved in the late steps of CT biosynthesis. Four days after treatment of Q. ilex seedlings by mechanical defoliation, P. cinnamomi infection and both stressors simultaneously, mRNA concentrations for tannins biosynthesis enzymes were measured in leaves. Among the transcript amount for shikimate dehydrogenase (SDH, EC 1.1.1.25), anthocyanidin reductase (EC 1.3.1.77), anthocyanidin synthase (EC 1.14.11.19) and leucoanthocyanidine reductase (EC 1.17.1.3), defoliation induced gene expression for SDH2 isoenzyme. About 4 days after infection of roots by P. cinnamomi, this up-regulation was canceled and SDH enzyme activity decreased. Furthermore, during this late stage of biotrophic interaction the pathogen switched off the correlation engaged by defoliation between the expression of SDH1 and SDH2 encoding genes and chemical defenses corresponding to total tannins, which were down-regulated. Thus, tannins biosynthesis in seedlings of Q. ilex is induced after mechanical defoliation whereas infection by the pathogen interferes with this regulation, potentially increasing the susceptibility of plants to herbivory and aggravating the impact of biotic stress.

Interaction of a Preventative Fungicide Treatment and Root Rot Pathogen on Ambrosia Beetle Attacks during a Simulated Flood Event.

Flooding can increase tree susceptibility to root rot pathogens as well as attacks by ambrosia beetles attracted to stress-induced ethanol emissions. The objective of this study was to investigate the interaction of a preventative fungicide treatment and root infection with Phytophthora cinnamomi on ambrosia beetle attacks in flood stressed trees. A fungicide (Pageant® Intrinsic®) was evaluated in two flood trials using Eastern redbud and tulip poplar trees with treatments including the fungicide with or without pathogen or no fungicide with or without pathogen. Fungicide treated trees had fewer ambrosia beetle attacks, particularly in trees without P. cinnamomi co-infection. In a follow-up experiment, ethanol content was evaluated in flooded redbuds to determine if the fungicide treatment reduced stress-induced compounds. All flood stressed trees began producing ethanol within 24 h post flooding, regardless of fungicide treatment or P. cinnamomi infection. We conclude that pre-treatments of a fungicide can provide protection from ambrosia beetle attacks during an extreme flood event, but that protection is reduced if a root rot pathogen is also present. Additionally, rejection of fungicide treated trees was not related to the absence of ethanol, as the fungicide-treated plants released ethanol in quantities similar to non-treated trees.

Relative importance of site, weather and Phytophthora cinnamomi in the decline and death of Eucalyptus marginata – jarrah dieback investigations in the 1970s to 1990s

Jarrah dieback was the name given to the sudden death of Eucalyptus marginata in the southwest of Western Australia, a serious economic problem. Although deaths were attributed to Phytophthora cinnamomi in the 1960s, the supporting evidence was weak; these deficiencies were not realised until 1980. Renewed interest in jarrah pathology showed that the incidence and severity of root lesions caused by P. cinnamomi in live trees was low, but in recent deaths it could be isolated from the root collar and large roots of some, but not all trees. Jarrah deaths result from hydraulic failure, implying extensive sapwood damage. This is unlikely to result from P. cinnamomi infection, which preferentially invades phloem, but could result from waterlogging, which causes tyloses to form in xylem vessels so they no longer conduct water. Tylosed root sapwood has been reported from investigations into jarrah deaths. An interpretation of past deaths based on stress factors better fits where and when deaths occur. This is within 3 years of exceptionally heavy rainfall, an inciting factor. Predisposing conditions are sites with some form of poor drainage, such as water-gaining sites, or those with impeded sub-soil drainage. Recent logging further increases site wetness. Phytophthora cinnamomi should be seen as a contributing factor, which is normally compartmentalised by the host, but can spread extensively in dying trees.

Generation of composite Persea americana (Mill.) (avocado) plants: A proof-of-concept-study.

Avocado (Persea americana (Mill.)), an important commercial fruit, is severely affected by Phytophthora Root Rot in areas where the pathogen is prevalent. However, advances in molecular research are hindered by the lack of a high-throughput transient transformation system in this non-model plant. In this study, a proof-of-concept is demonstrated by the successful application of Agrobacterium rhizogenes-mediated plant transformation to produce composite avocado plants. Two ex vitro strategies were assessed on two avocado genotypes (Itzamna and A0.74): In the first approach, 8-week-old etiolated seedlings were scarred with a sterile hacksaw blade at the base of the shoot, and in the second, inch-long incisions were made at the base of the shoot (20-week-old non-etiolated plants) with a sterile blade to remove the cortical tissue. The scarred/wounded shoot surfaces were treated with A. rhizogenes strains (K599 or ARqua1) transformed with or without binary plant transformation vectors pRedRootII (DsRed1 marker), pBYR2e1-GFP (GFP- green fluorescence protein marker) or pBINUbiGUSint (GUS- beta-glucuronidase marker) with and without rooting hormone (Dip 'N' Grow) application. The treated shoot regions were air-layered with sterile moist cocopeat to induce root formation. Results showed that hormone application significantly increased root induction, while Agrobacterium-only treatments resulted in very few roots. Combination treatments of hormone+Agrobacterium (-/+ plasmids) showed no significant difference. Only the ARqua1(+plasmid):A0.74 combination resulted in root transformants, with hormone+ARqua1(+pBINUbiGUSint) being the most effective treatment with ~17 and 25% composite plants resulting from strategy-1 and strategy-2, respectively. GUS- and GFP-expressing roots accounted for less than 4 and ~11%, respectively, of the total roots/treatment/avocado genotype. The average number of transgenic roots on the composite plants was less than one per plant in all treatments. PCR and Southern analysis further confirmed the transgenic nature of the roots expressing the screenable marker genes. Transgenic roots showed hyper-branching compared to the wild-type roots but this had no impact on Phytophthora cinnamomi infection. There was no difference in pathogen load 7-days-post inoculation between transformed and control roots. Strategy-2 involving A0.74:ARqua1 combination was the best ex vitro approach in producing composite avocado plants. The approach followed in this proof-of-concept study needs further optimisation involving multiple avocado genotypes and A. rhizogenes strains to achieve enhanced root transformation efficiencies, which would then serve as an effective high-throughput tool in the functional screening of host and pathogen genes to improve our understanding of the avocado-P. cinnamomi interaction.

Variation in susceptibility among macadamia genotypes and species to Phytophthora root decay caused by Phytophthora cinnamomi

Phytophthora cinnamomi is a major pathogen of cultivated macadamia (Macadamia integrifolia, Macadamia tetraphylla and their hybrids) worldwide. The susceptibility of the two non-edible Macadamia species (Macadamia ternifolia and Macadamia jansenii) to P. cinnamomi is not well-understood. Commercial macadamia trees are established on grafted seedling (seed propagation) or own-rooted cutting (vegetative propagation) rootstocks of hybrids of the cultivated species. There is little information to support the preferential use of rootstock propagated by either seedling or own-rooted cutting methods in macadamia. In this study we assessed roots of macadamia plants of the four species and their hybrids, derived from the two methods of propagation, for their susceptibility to P. cinnamomi infection. The roots of inoculated plant from which P. cinnamomi was recovered showed blackening symptoms. The non-cultivated species, M. ternifolia and M. jansenii and their hybrids were the most susceptible germplasm compared with M. tetraphylla and M. integrifolia. Of these two species, M. tetraphylla was less susceptible than M. integrifolia. Significant differences were observed among the accessions of their hybrids. A strong association (R2 > 0.75) was recorded between symptomatic roots and disease severity. Root density reduced with increasing disease severity rating in both own-rooted cuttings (R2 = 0.65) and germinated seedlings (R2 = 0.55). P. cinnamomi severity data were not significantly (P > 0.05) different between the two methods of plant propagation. The significance of this study to macadamia breeding and selection of disease resistant rootstocks is discussed.

Dual RNA-Sequencing of Eucalyptus nitens during Phytophthora cinnamomi Challenge Reveals Pathogen and Host Factors Influencing Compatibility.

Damage caused by Phytophthora cinnamomi Rands remains an important concern on forest tree species. The pathogen causes root and collar rot, stem cankers, and dieback of various economically important Eucalyptus spp. In South Africa, susceptible cold tolerant Eucalyptus plantations have been affected by various Phytophthora spp. with P. cinnamomi considered one of the most virulent. The molecular basis of this compatible interaction is poorly understood. In this study, susceptible Eucalyptus nitens plants were stem inoculated with P. cinnamomi and tissue was harvested five days post inoculation. Dual RNA-sequencing, a technique which allows the concurrent detection of both pathogen and host transcripts during infection, was performed. Approximately 1% of the reads mapped to the draft genome of P. cinnamomi while 78% of the reads mapped to the Eucalyptus grandis genome. The highest expressed P. cinnamomi gene in planta was a putative crinkler effector (CRN1). Phylogenetic analysis indicated the high similarity of this P. cinnamomi CRN1 to that of Phytophthora infestans. Some CRN effectors are known to target host nuclei to suppress defense. In the host, over 1400 genes were significantly differentially expressed in comparison to mock inoculated trees, including suites of pathogenesis related (PR) genes. In particular, a PR-9 peroxidase gene with a high similarity to a Carica papaya PR-9 ortholog previously shown to be suppressed upon infection by Phytophthora palmivora was down-regulated two-fold. This PR-9 gene may represent a cross-species effector target during P. cinnamomi infection. This study identified pathogenicity factors, potential manipulation targets, and attempted host defense mechanisms activated by E. nitens that contributed to the susceptible outcome of the interaction.

How Phytophthora cinnamomi became associated with the death of Eucalyptus marginata – the early investigations into jarrah dieback

The name jarrah dieback was used in the 1940s to describe a serious economic problem in the jarrah forest in the south west of Western Australia. This was the sudden death of groups of jarrah (Eucalyptus marginata) trees that occurred on previously logged sites that had a tendency to become waterlogged in winter. Although the cause was not determined at the time, from symptoms recorded in early investigations the most likely explanation is that the trees died as the result of waterlogging damage. In the 1960s it was shown that many of these sites were infested by the introduced oomycete Phytophthora cinnamomi and tree deaths, together with the deaths of many mid- and under-storey plants, were attributed to this pathogen. A chronology of the research, based on contemporary unpublished documents, shows that in 1968 the conclusion that P. cinnamomi caused jarrah deaths was not supported by the available evidence, because the work did not satisfy the first and fourth of Koch’s postulates. The evidence that P. cinnamomi killed many mid- and under-storey plants was much stronger. There are two problems that have been confused: the death of groups of jarrah trees (jarrah dieback) that is caused by waterlogging and the death of many mid- and under-storey plants (Phytophthora dieback) caused by P. cinnamomi infection.

Phylogenetic and expression analysis of the NPR1-like gene family from Persea americana (Mill.).

The NONEXPRESSOR OF PATHOGENESIS-RELATED GENES1 (NPR1) forms an integral part of the salicylic acid (SA) pathway in plants and is involved in cross-talk between the SA and jasmonic acid/ethylene (JA/ET) pathways. Therefore, NPR1 is essential to the effective response of plants to pathogens. Avocado (Persea americana) is a commercially important crop worldwide. Significant losses in production result from Phytophthora root rot, caused by the hemibiotroph, Phytophthora cinnamomi. This oomycete infects the feeder roots of avocado trees leading to an overall decline in health and eventual death. The interaction between avocado and P. cinnamomi is poorly understood and as such limited control strategies exist. Thus uncovering the role of NPR1 in avocado could provide novel insights into the avocado – P. cinnamomi interaction. A total of five NPR1-like sequences were identified. These sequences were annotated using FGENESH and a maximum-likelihood tree was constructed using 34 NPR1-like protein sequences from other plant species. The conserved protein domains and functional motifs of these sequences were predicted. Reverse transcription quantitative PCR was used to analyze the expression of the five NPR1-like sequences in the roots of avocado after treatment with salicylic and jasmonic acid, P. cinnamomi infection, across different tissues and in P. cinnamomi infected tolerant and susceptible rootstocks. Of the five NPR1-like sequences three have strong support for a defensive role while two are most likely involved in development. Significant differences in the expression profiles of these five NPR1-like genes were observed, assisting in functional classification. Understanding the interaction of avocado and P. cinnamomi is essential to developing new control strategies. This work enables further classification of these genes by means of functional annotation and is a crucial step in understanding the role of NPR1 during P. cinnamomi infection.

Quantitative RT-PCR analysis of differentially expressed genes in Quercus suber in response to Phytophthora cinnamomi infection.

cDNA-AFLP methodology was used to gain insight into gene fragments differentially present in the mRNA profiles of Quercus suber roots infected with zoospores of Phytophthora cinnamomi at different post challenge time points. Fifty-three transcript-derived fragments (TDFs) were identified and sequenced. Six candidate genes were selected based on their expression patterns and homology to genes known to play a role in defence. They encode a cinnamyl alcohol dehydrogenase2 (QsCAD2), a protein disulphide isomerase (QsPDI), a CC-NBS-LRR resistance protein (QsRPc), a thaumatin-like protein (QsTLP), a chitinase (QsCHI) and a 1,3-β-glucanase (QsGlu). Evaluation of the expression of these genes by quantitative polymerase chain reaction (qPCR) revealed that transcript levels of QsRPc, QsCHI, QsCAD2 and QsPDI increased during the first 24 h post-inoculation, while those of thaumatin-like protein decreased. No differential expression was observed for 1,3-β-glucanase (QsGlu).Four candidate reference genes, polymerase II (QsRPII), eukaryotic translation initiation factor 5A (QsEIF-5A), β-tubulin (QsTUB) and a medium subunit family protein of clathrin adaptor complexes (QsCACs) were assessed to determine the most stable internal references for qRT-PCR normalization in the Phytophthora-Q. suber pathosystem in root tissues. Those found to be more stable, QsRPII and QsCACs, were used as internal reference in the present work.Knowledge on the Quercus defence mechanisms against biotic stress is scarce. This study provides an insight into the gene profiling of a few important genes of Q. suber in response to P. cinnamomi infection contributing to the knowledge of the molecular interactions involving Quercus and root pathogens that can be useful in the future to understand the mechanisms underlying oak resistance to soil-borne oomycetes.Electronic supplementary materialThe online version of this article (doi:10.1186/2193-1801-3-613) contains supplementary material, which is available to authorized users.

Fine root dynamics of oak saplings in response to Phytophthora cinnamomi infection under different temperatures and durations

SummaryThe belowground effects of Phytophthora cinnamomi on 1‐year‐old saplings of two common oak species in mid‐Atlantic US forests, white (Quercus alba) and red oak (Q. rubra), were examined after incubation in pathogen‐infested soilless potting mix. Fine root lengths (0–1.5 mm in diameter) of both oak species were quantified after incubation at successive 30‐day intervals up to 300 days, for a total of 10 incubation periods. In addition, colony‐forming units (CFU) of P. cinnamomi were quantified after white oak saplings were incubated in infested soilless potting mix at different temperature/duration combinations that reflect soil conditions present in the mid‐Atlantic United States. Impact of P. cinnamomi on fine root lengths of red and white oak saplings varied considerably over time. Significant periods of fine root loss occurred primarily during spring, when bud break and leaf flush began for both oak species. Red oaks had 17% fine root loss on average, while white oaks appeared more resistant to P. cinnamomi infection with a 2% decrease in fine roots over the course of the experiment. Phytophthora cinnamomi CFU declined significantly with exposure to all incubation temperatures except 8°C. This was in contrast to in vitro experiments, where the optimum temperature for mycelial growth was determined to be 21°C and above. Significant fine root loss caused by P. cinnamomi depended on plant phenology and the oak species tested. Extreme soil temperatures have a significant adverse impact on temporal changes of P. cinnamomi population.

Composted Cotton Stalks and Cotton Gin Trash Substrate Amendments and Irrigation/Ground Cover Management II. Effect on Growth and Disease Suppression for Azalea and Juniper

‘Sunglow’ azalea and ‘Blue Pacific’ juniper were grown in pine bark (PB) and pine tree (PT) substrates that were amended with cotton stalks composted with a N source (CSN), cotton stalks composted without an N source (CS), and cotton gin trash (CGT) to evaluate the substrate's effect on plant growth and disease suppression. The plants were grown under two different, commonly used, irrigation/ground surface management regimes — overhead, sprinkler irrigation with black geotextile weed fabric covering the ground (OH) or low-volume, spray stake irrigation with gravel covering the ground (LV). In 2010, with OH, all PB-amended substrates produced significantly larger azalea shoots than PT-amended substrates. In 2011, with OH, all azalea shoots were similar in size when grown in all substrates except for PT:CS, where plants were significantly smaller. With LV, in 2010 and 2011, azalea shoot growth was largest when grown in a PB substrate amended with CSN or CGT and lowest in PT:CS. Junipers with OH produced generally larger shoot growth with the PB-based substrates in both 2010 and 2011 compared to the PT-based substrates. With LV, PT:CGT produced the numerically smallest juniper shoot growth for both years. Overall, PT-based substrates appeared to produce greater consistency in growth, because responses were more similar in 2010 and 2011, however irrigation method and management can impact growth regardless of substrate composition. CGT added to PB- or PT-based substrates enhanced Ca and Mg uptake by both species but not P uptake. OH generally kept ground surface and substrate temperatures lower than LV regardless of substrate composition. The substrates tested neither enhanced nor deterred P. cinnamomi infection in azalea or juniper.

Drought events determine performance of Quercus ilex seedlings and increase their susceptibility to Phytophthora cinnamomi

More frequent weather extremes are expected to occur in the Mediterranean region within the present context of climate change. These extremes could affect forests and plant diseases driven by pathogens. It is hypothesised that simulation of weather extremes during Quercus ilex growth will influence early performance and susceptibility to the invasive oomycete Phytophthora cinnamomi. In 2010, 140 Q. ilex seedlings were subjected to three watering regimes under greenhouse conditions: waterlogging (W), water stress (S) and optimal watering regime for growth (C). During the second vegetative period, conditions were altered to create the following scenarios: WW, WS, SS, SW and CC. After the second vegetative period, plants were artificially infested with P. cinnamomi. Holm oak (Q. ilex) was more sensitive to flooding in the first year of growth than in the second year. The altered scenarios produced plants with a lower fine-to-total root ratio than plants in unaltered scenarios. Plants with the highest growth rates maintained their relatively rapid growth and photosynthetic activity under altered scenarios. However, plants with the highest growth rates became the plants with the lowest growth rates when two consecutive years of drought occurred, indicating a trade-off by Q. ilex in growth investment, observed only if the water stress scenario persists. Seedlings were more sensitive to water shortage than to waterlogging, especially if they encountered a dry scenario during the first year. Exposure to drought events increased seedling mortality rates after P. cinnamomi infection. Waterlogging combined with subsequent water deprivation was the worst scenario when soil was infested with P. cinnamomi, causing 100% mortality of plants.

Anti-Phytophthora cinnamomi activity of Phlomis purpurea plant and root extracts

Phlomis purpurea (Lamiaceae), found in Quercus suber and Quercus ilex ssp. rotundifolia forest habitats in southern Portugal, is a non-host for the oomycete Phytophthora cinnamomi, the main biotic factor involved in cork oak and holm oak decline in the Iberian Peninsula. The effect of P. purpurea crude ethanol root extract was evaluated in vitro on P. cinnamomi mycelial growth, sporangial production, zoospore release and germination as well as on chlamydospore production and viability. The protection of cork oak against infection by the pathogen was also evaluated in planta. At 10 mg ml−1, in vitro inhibition of the pathogen structures was 85–100 %. In addition, P. purpurea plants were shown to protect Q. suber and Q. ilex from P. cinnamomi infection and to reduce the inoculum potential in glasshouse trials, indicating the ability to reduce root infection by the pathogen. The results suggest that P. purpurea has the potential to reduce disease spread and that their root extracts could provide candidate substances for control of the important pathogen, P. cinnamomi.

Variation in susceptibility of Banksia (including Dryandra) to Phytophthora cinnamomi

Variation in susceptibility of 109 Western Australian Banksia and Dryandra and 16 Eastern Australian Banksia to Phytophthora cinnamomi infection was evaluated after soil inoculation. Banksia and Dryandra were initially analysed separately and then combined due to the transfer of Dryandra to Banksia. Disease progress curves of % mortality following soil inoculation were analysed with the three logistic model parameters of upper asymptote (Kmax), lag time (t½K) and intrinsic rate of increase (r). The susceptibility continuum represented by the ordination of parameters of the logistic model for % plant mortality in two dimensions was similar for Banksia, Dryandra and combined Banksia and Dryandra. Although resistant taxa had low Kmax, the values for delay and rate varied. In comparison, susceptible taxa tended to have high Kmax short delay and fast rate. Western Australian threatened and priority Banksia were significantly more susceptible than common Banksia. Seeders were more susceptible than resprouters. There was no significant difference in Western Australian Banksia susceptibility between soil, rainfall region, bioregion and growth form. Of the Western Australian Banksia tested, 40 % were most at risk of P. cinnamomi infection because they were either threatened, priority or common though geographically restricted taxa with Kmax of mortality >50 %. Positioning Banksia taxa on the P. cinnamomi resistance-susceptibility continuum needs to be incorporated into extinction-risk methodology in order to prioritise flora for conservation actions according to hazard from the pathogen.

Cryptogein and capsicein promote defence responses in Quercus suber against Phytophthora cinnamomi infection

The decline of cork oak (Quercus suber) stands in Iberian Peninsula is associated with infection by Phytophthora cinnamomi. Most Phytophthora spe- cies secrete elicitins, which can enhance defence reac- tions against some pathogens. Here cytological and physiological effects of the elicitins cryptogein and capsicein on cork oak root infection by P. cinnamomi were evaluated. The progression of the pathogen in root tissue and its effects on total fatty acid (TFA) composition of roots and leaves were analysed in seedlings. Net photosynthesis (Pn), stomatal conductance (gs), chlorophyll a fluorescence measure- ments (quantum yield of linear electron transport ϕe, photochemical quenching qP, non-photochemical quenching NPQ) and carotenoid determinations were carried out in well established (4 months) plants. In elicitin-treated roots, 2 days after inoculation, the path- ogen which presented loss of viability and membrane degradation was mainly restricted to the intercellular spaces of the cortical parenchyma, and did not reach the vascular cylinder. Electron dense materials accumu- lated in the intercellular spaces of the cortex next to disorganized hyphae, suggested to be related with de- fence reactions. Cryptogein (or its interaction with P. cinnamomi) induced enhanced lipid synthesis in leaves, which may contribute to preserve membrane stability. P. cinnamomidecreasedPn,gs, ϕe,andqP,whereaselicitin- treated plants displayed values similar to controls. Over- all, the results indicated a resistance response of cork oak against this oomycete, induced by the elicitins.

Variation in susceptibility to Phytophthora cinnamomi infection within the genus Lambertia

Variation in susceptibility to Phytophthora cinnamomi Rands infection within the genus Lambertia Sm. was evaluated by soil and stem inoculation. Disease-progress curves following soil inoculation were analysed with the three logistic-model parameters of upper asymptote (K max), lag time (t 1/2K ) and intrinsic rate of increase (r), which were combined into a mortality score. Resistant Lambertia taxa had the lowest K max, longest t 1/2K and slowest r. These parameters change to greatest K max, shortest t 1/2K and fastest r for susceptible Lambertia taxa. Mortality score following soil inoculation was significantly positively correlated with lesion score determined by stem inoculation. The plot of the scores ranked the Lambertia taxa in relation to P. cinnamomi susceptibility on the resistance–susceptibility continuum and prioritised taxa in relation to the threat posed by the pathogen. The highest mortality and lesion scores for the threatened rare and endangered taxa L. orbifolia subsp. orbifolia, L. fairallii and L. rariflora subsp. lutea suggest a high risk of extinction from P. cinnamomi infestation. Furthermore, rare and endangered taxa L. orbifolia subsp. Scott River Plains, L. echinata subsp. occidentalis and L. echinata subsp. echinata that have high mortality and moderate lesion scores are also likely to be at high risk of extinction from P. cinnamomi infestation. Among common taxa with restricted geographic distribution, L. ericifolia has high mortality and lesion scores, suggesting a high risk of localised extinction and loss of genetic diversity in P. cinnamomi disease centres. Positioning taxa on the P. cinnamomi resistance–susceptibility continuum needs to be incorporated into extinction-risk methodology so as to prioritise flora for conservation actions according to hazard from the pathogen.