Occurrence and distribution of <Emphasis Type="Italic">Phytophthora</Emphasis> species in European chestnut stands,and their association with Ink Disease and crown decline

The Phytophthora complex associated with Castanea sativa Mill. was investigated in five European countries in 35 regions and with respect to various domestication levels. Annual precipitation and length of drought season were the main parameters that regulated the presence of Phytophthora species in the chestnut stands. Seven species of Phytophthora were detected; three of these, P. megasperma, P. cryptogea and P. syringae had not been previously reported on sweet chestnut. P. cinnamomi. P. cambivora and P. citricola were most frequently isolated. P. cinnamomi and P. cambivora were the species significantly associated with declining trees with symptoms of Ink Disease. P. cinnamomirequired distinct ecological conditions compared to the other species. P. cinnamomi was never detected in sites characterized by minimum temperatures below 1.4 °C, maximum temperature above 28 °C, or soil pH below 5.4. The results obtained provide useful information for modeling the probability of Ink Disease, crown decline and associated Phytophthora species in chestnut groves in global climatic change scenarios.     

Diagnostic sensitivity and specificity of different methods used by two laboratories for the detection of Phytophthora ramorum on multiple natural hosts

Five detection methods were comparatively tested on putative Phytophthora ramorum field samples from 41 wild plant species. The tested methods included two culture‐based assays, a DAS‐ELISA‐based polyclonal assay, a nested PCR‐based assay, and a TaqMan real‐time PCR assay. Diagnostic values including sensitivity, specificity, positive predictive value and negative predictive value were calculated for each method. The effects of host species, seasonality and host location were analysed and compared between two laboratories. Significant effects of season, host species and laboratory were detected. It is concluded that a combination of either culturing and molecular diagnosis or of two molecular assays is the most promising approach to diagnose this pathogen. Based on the results of this and other studies, diagnosis should occur as much as possible during wet and warm periods favourable to the pathogen, and proficiency tests should be performed to compare results obtained with molecular approaches in different laboratories. Furthermore, length of time lapsed between sample collection and processing strongly affected the diagnostic sensitivity of culture‐based methods, and therefore needs to be taken into account when comparing results from different laboratories.     

Evaluation of root damage to English walnut caused by five Phytophthora species

The pathogenicity of five species of Phytophthora to English walnut was studied in a greenhouse experiment. Phytophthora cinnamomi was the most aggressive species, causing severe root rot and seedling mortality. The other species tested, P. cambivora , P. citricola , P. cactorum and P. cryptogea , did not induce visible crown symptoms on seedlings 2 months after inoculation. Some strains of P. cambivora and P. cactorum also caused taproot damage to seedlings. All except one of the tested isolates caused significant necrosis of fine roots and a significant reduction of root weight compared with noninoculated seedlings. Reduction of above-ground plant development was not statistically significant. While P. cinnamomi is well known as an aggressive primary pathogen of English walnut, the other species of Phytophthora may act as predisposing factors to walnut decline, affecting root system development and increasing host vulnerability to environmental stress.     

Occurrence of Phytophthora cinnamomi in cork oak forests in Italy

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.     

Phytophthora lateralis discovered in an old growth Chamaecyparis forest in Taiwan

The geographic origins of the invasive Phytophthora species, P. lateralis and P. ramorum are unknown. In 2008 soil samples were collected in an old growth yellow cedar (Chamaecyparis obtusa var. formosana) stand in the Ma‐kau Ecological Park in north eastern Taiwan and subjected to Phytophthora baiting procedures at 18°C. Cedar needle baits yielded isolates of a slow growing Phytophthora culture from one soil sample, together with P. cinnamomi. Phytophthora bisheria sp. nov. was obtained from another sample. The slow growing isolates conformed closely to P. lateralis in the morphology of their sporangia and chlamydospores, growth–temperature relationships, absence of gametangia and their ITS and cox II sequences. The isolates’ sporangia were partially caducous, with short pre‐formed pedicels of ca. 3–5 μm, a highly unusual feature in a non‐papillate Phytophthora. The isolates also produced multicellular stromata on cedar decoction agar. Small morphological and molecular differences were observed between the Taiwan‐isolates and Oregon‐control isolates. Taiwan may lie within the geographic centre of origin of P. lateralis. By analogy Japan may also lie within the natural range of P. lateralis; and Japan, along with Taiwan and Yunnan, could be an origin for the closely related P. ramorum.     

Behaviour and survival of Phytophthora cambivora inoculum in soil‐like substrate under different water regimes

Phytophthora cambivora is a soil‐borne pathogen responsible for root and collar rot of woody species including Castanea spp., on which it causes Ink disease. P. cambivora does not produce chlamydospores, and the prevalence of a single mating type in nature limits the production of oogonia and oospores. Thus, survival of P. cambivora, in the absence of suitable hosts, relies mainly on sporangia, zoospores and mycelium. However, inoculum survival in soils or substrates represents a major factor in disease epidemics and is probably dependant on environmental factors. The aim of this article was to study the behaviour of P. cambivora in a peat‐based substrate (PbS) in the absence of a host, under controlled conditions, and by varying the substrate water regimes. Total inoculum in the substrate was quantified by qPCR, using customized rDNA primer sets, whilst zoospores were quantified after elution, and their vitality was determined by a specific staining procedure. Inoculum infectivity was assessed using chestnut root baits. Results indicated that during the initial 48 h after infestation (matric potential, pF 1.2; 70% water content), the inoculum level increased producing microsporangia and mycelium before decreasing, regardless of the water regime applied. The number of free zoospores increased over a period of (maximum) 7 days after substrate inoculation. Vitality of zoospores declined to 45% at day 18, with no statistical differences amongst water regime treatments. Both rDNA and zoospore number increased in flooded samples at day 23, illustrating the vitality of the inoculum. Inoculum was able to infect chestnut root baits up to 45 days at pF 1.2 and 3.5, before and after flooding, but at pF 6.3, only in the absence of flooding. Although based on experiments carried out in microcosms, these results highlight an unexpected resistance of P. cambivora zoospores to environmental constraints. Furthermore, the infectivity ability of the inoculum to survive in PbS over time even in the absence of the host adds relevance to the risk of movement of soil and substrates associated with plant trading worldwide.     

First report of the ambrosia beetle Xylosandrus compactus and associated fungi in the Mediterranean maquis in Italy,and new host–pest associations

In September 2016, a survey conducted in the Circeo National Park revealed an outbreak and serious damage caused by the black twig borer (Xylosandrus compactus) and its associated fungi in the Mediterranean maquis. Among the affected hosts, Quercus ilex, Viburnum tinus, Ruscus aculeatus, Pistacia lentiscus, Laurus nobilis and Ceratonia siliqua, showed flagging and wilting of branches and, in younger individuals, the death of the whole plant occurred. In total, 18 different fungal taxa were found associated with the insect. These included Ambrosiella xylebori, Geosmithia pallida, Fusarium spp., Epicoccum nigrum and Bionectria sp. This is the first report in Europe of X. compactus and associated ambrosia fungi in a natural environment.     

Characterization of Phytophthora cinnamomi from common walnut in Southern Europe environment

Forty‐eight isolates of Phytophthora cinnamomi obtained from common walnut were analysed according to their variability in growth at different temperatures, virulence, sensitivity to metalaxyl and in genomic DNA. Isolates were obtained from commercial common walnut orchards located in northern Italy and in southern France. Inter‐simple sequence repeat (ISSR) were analysed for the 49 isolates, 43 of which were Italian, 6 French; an isolate of the same species obtained from Viburnum spp. was used as an outgroup. ANOVA on phenotypic characters showed a significant impact of the geographic location of the orchard on isolate variability in terms of reaction to temperatures and aggressiveness. In turn, clustering obtained with UPGMA analysis on genetic data was almost exclusively dependant on isolate variability, nevertheless the 48 isolates seem to share a common variability that differentiates the group from the isolate from Viburnum spp. Correlation between phenotypic and genetic traits was not statistically significant. In conclusion, phenotypic variability like virulence seemed to be conditioned from geographic origin while the genetic variability of P. cinnamomi isolates from walnut was associated to the single genotype.     

Detection,description, and technological properties of colour aberration in wood of standards and shoots from a chestnut (Castanea sativa Mill.) coppice stand

European Journal of Forest Research - Sweet chestnut forests occupy an important role in Italy and Europe, its resilience is often affected by aggressive pathogen attack. In the last times some...     

The morphology,behaviour and molecular phylogeny of Phytophthora taxon Salixsoil and its redesignation as Phytophthora lacustris sp. nov.

Since its first isolation from Salix roots in 1972, isolates of a sexually sterile Phytophthora species have been obtained frequently from wet or riparian habitats worldwide and have also been isolated from roots of Alnus and Prunus spp. Although originally assigned to Phytophthora gonapodyides on morphological grounds, it was recognized that these isolates, informally named P. taxon Salixsoil, might represent a separate lineage within ITS Clade 6. Based on phylogenetic analyses and comparisons of morphology, growth‐temperature relationships and pathogenicity, this taxon is formally described here as Phytophthora lacustris sp. nov. Isolates of P. lacustris form a clearly resolved cluster in both ITS and mitochondrial cox1 phylogenies, basal to most other Clade 6 taxa. Phytophthora lacustris shares several unusual behavioural properties with other aquatic Clade 6 species, such as sexual sterility and tolerance of high temperatures, that have been suggested as adaptations to riparian conditions. It appears to be widespread in Europe and has also been detected in Australia, New Zealand and the USA. It was shown to be weakly or moderately aggressive on inoculation to Alnus, Prunus and Salix. The extent of P. lacustris’ activity as a saprotroph in plant debris in water and as an opportunistic pathogen in riparian habitats needs further investigation. Its pathogenic potential to cultivated fruit trees also deserves attention because P. lacustris has apparently been introduced into the nursery trade.