Pestalotiopsis Funerea Research Articles

Antifungal Potential and Chemical Composition of Chamaecyparis formosensis and Chamaecyparis obtusa var. formosana Essential Oils in Liquid and Vapor Phases Against 4 Fungal Species

Background Only a few studies have investigated the bioactivity of the essential oils (EOs) of Chamaecyparis formosensis and C. obtusa var. formosana, which are endemic to Taiwan. The wood and leaf EOs extracted from C. obtusa var. formosana had anti-inflammatory and antitermitic effects, respectively, and the twig EO extracted from C. formosensis inhibited the growth of Fusarium oxysporum, Pestalotiopsis funerea, and Ganoderma australe. Methods A poisoned food technique was used to evaluate the antifungal effects of wood and leaf EOs extracted from C. formosensis and C. obtusa var. formosana on brown rot fungus, white rot fungus, and mold. A phytatray chamber assay was conducted to investigate the antifungal activities of the EOs in the vapor phase. Gas chromatography-mass spectrometry was used to examine the chemical constituents of both EOs. Results The leaf EO extracted from C. formosensis primarily contained monoterpene hydrocarbons, whereas the leaf EO extracted from C. obtusa var. formosana primarily contained monoterpene and sesquiterpene hydrocarbons. The wood EOs extracted from C. formosensis and C. obtusa var. formosana primarily contained sesquiterpene hydrocarbons and oxygenated sesquiterpenes. These results indicated that the wood EOs had higher antifungal activity in the liquid phase, whereas the leaf EOs had higher antifungal activity in the vapor phase. Conclusion Wood EOs extracted from C. formosensis and C. obtusa var. formosana have strong antifungal effects against Gloeophyllum trabeum, Lenzites betulina, Trametes versicolor, and Trichoderma sp. Compared with wood EOs, leaf EOs have a greater inhibitory effect on mycelial growth in the vapor phase. In both the liquid and vapor phases, the EOs extracted from C. formosensis and C. obtusa var. formosana serve as effective antifungal agents against pathogenic fungi. Further research is required to elucidate the mechanisms underlying the antifungal activities of EOs in different phases and explore their potential applications.

Türkiye’nin kuzeybatısındaki bir plantasyonda yerli ve egzotik çam türlerinin ibrelerinde görülen fungal endofitler

The endophytic fungi present in needles of Pinus sylvestris, Pinus pinaster, Pinus nigra, Pinus taeda and Pinus radiata were investigated in Kerpe Research Forest, İzmit in 2016. Ten trees of each pine species were sampled systematically. Previous years green needles were sampled from the lower part of the canopy, from two equally spaced positions around the tree. Each needle was surface sterilized and cut into 0.5 cm sections then individually placed onto malt extract agar plates. DNA was ex-tracted from reprensentative seven isolates and amplified using primers ITS1 and ITS4 targeting the nuclear 5.8S rDNA gene and the two ITS regions flanked between 18S and 28S rDNA genes. Amplicons sequenced in both directions using the univer-sal fungal primers ITS1 and ITS4. Isolations from a total of 1000 needles (200 from each pine species) yielded 750 fungal isolates. Pestalotiopsis funerea (Desm.) Steyaert was isolated at the highest frequency followed by Acremonium sp., Cladio-sporum sp. and Cyclaneusma minus.

Blueberry mycobiota

   We have identified the following pathogenic fungi on blueberries: Godronia casandra pck = Fusicocum putrefaciens Sheare; Verticillium dahliae kleeb; Fusarium oxysporium (Schlecht) Snyd. et Haans.; Pestalotiopsis guepini (Desm) Steyaert.; Pestalotiopsis neglecta (Thumen) Steyaert.; Pestalotiopsis funerea (Desm) Steyaert.; Alternaria alternata(Fr.) Keisl. and Botrytinia fuckeliana(De Bary)Whetzel = Botrytis cinerea Pers. Godronia casandra pck = Fusicocum putrefaciens Sheare was noted by us for the first time in Georgia in 2020 on seedlings and 3-4 year old plants imported from abroad. It is soil-dwelling, highly pathogenic, quarantined. Fungi with great pathogenic nature are also: Verticillium dahliae kleeb and Fusarium oxysporium (Schlecht)Snyd. et Haans. which cause wilting of plants, falling of leaves and withering of stems and branches.

Investigations on Fungi Isolated from Apple Trees with Die-Back Symptoms from Basilicata Region (Southern Italy).

Val d’Agri is an important orchard area located in the Basilicata Region (Southern Italy). A phenomenon affecting cv. “Golden Delicious” apples which lead to tree death has been observed in the past several years in this area. This phenomenon has already been detected in about 20 hectares and is rapidly expanding. The symptoms observed were “scaly bark” and extensive cankers, mainly located in the lower part of the trunk, associated with wood decay. Dead plants ranged from 20% to 80% and, in many cases, trees were removed by farmers. In order to identify the causes of this phenomenon, investigations were started in autumn/winter 2019. In order to determine the possible causal agents, fungal and bacterial isolations, from symptomatic tissues, were performed in laboratory. Bacterial isolations gave negative results, whereas pure fungal cultures (PFCs) were obtained after 3–4 passages on potato dextrose agar (PDA) media. Genetic material was extracted from each PFC and amplified by PCR using three pairs of primers: ITS5/4, Bt2a/Bt2b and ACT-512F/ACT-783R. The amplicons were directly sequenced, and nucleotide sequences were compared with those already present in the NCBI GenBank nucleotide database. All isolated fungi were identified based on morphological features and multilocus molecular analyses. Neofusicoccum parvum, Diaporthe eres and Trametes versicolor were most frequently isolated, while Pestalotiopsis funerea, Phomopsis spp. and Diaporthe foeniculina were less frequently isolated. All nucleotide sequences obtained in this study have been deposited into the EMBL database. Pathogenicity tests showed that N. parvum was the most pathogenic and aggressive fungus, while Phomopsis sp. was demonstrated to be the less virulent one. All the investigated fungi were repeatedly reisolated from artificially inoculated twigs of 2-year-old apple trees, cv. “Golden Delicious”, and subsequently morphologically and molecularly identified. The role played by the above-mentioned fungi in the alterations observed in field is also discussed.

Foliar endophytes in trees varying greatly in age

Temple trees, including the gymnosperm Platycladus orientalis and the angiosperm Styphnolobium japonicum, have been planted in China for thousands of years. Tree age thus varies widely from young to ancient trees. Foliar endophytes of P. orientalis and S. japonicum were surveyed in this exploratory study that was based on isolation into culture and sequencing of fungi from trees varying in age from 10 to 5000 years (P. orientalis) and from 10 to 1700 years (S. japonicum). Sequenced endophytes of P. orientalis and S. japonicum belonged to 24 and 16 fungal genera, respectively. Principal components analysis showed that 14 components were necessary to explain 90% of the variance in endophyte community structure in P. orientalis. In S. japonicum eight components were needed for 90%. It is against that backdrop of complex etiology in community structuring, that the relative frequencies (abundances) of 17 of the 24 endophytes from P. orientalis and 9 of the 16 from S. japonicum were significantly correlated with tree age. There were two major trends. Abundant fungal genera [Fusarium + Alternaria = 74.57% (P. orientalis) and 81.24% (S. japonicum)] tended to decline linearly with tree age. Most of the rare fungal genera, in contrast, increased in relative abundance linearly with tree age. Diversity (H′) and richness (Margalef) of endophyte communities in foliage thus increased as the trees aged. Relative abundances of pathogenic endophytes, or latent pathogens, (Pestalotiopsis funerea and Amyloporia subxantha in P. orientalis; Collectotrichum gloeosporioides and Botryosphaeria dothidea in S. japonicum) also increased linearly as the trees aged.Since leaf age does not vary with tree age in the deciduous S. japonicum, nor in the evergreen P. orientalis, ‘tree age’ currently lacks a mechanistic explanation for its apparent importance among common foliar endophytes.

Determination of chemical compositions and antifungal activities of laurel and fennel essential oils against fungal disease agents of cypress seedlings

Fusarium oxysporum and Pestalotiopsis funerea are the most common fungal disease agents of conifer seedlings causing root rot and shoot or tip blight diseases. In this study, chemical compositions and antifungal activities of essential oils of fennel (Foeniculum vulgare Mill.) and laurel (Laurus nobilis L.) were determined against root rot and wilt disease agents F. oxysporum and P. funerea in vitro conditions. Chemical compositions of essential oils were determined by using GC-MS analysis. Antifungal volatile phase effects of essential oils were determined on inhibition of mycelial growth in vitro conditions by using different concentrations. The effect of most effective concentrations of essential oils on the morphology of fungal hypha was also determined by using light microscope. GC-MS analysis of essential oils of laurel and fennel plants revealed that eucalyptol (46.97%) and α-terpinyl acetate (19.82%) were major components of laurel; trans-anethole (82.44%) and limonene (5.18%) were major components of fennel essential oils. Volatile phase effects of fennel and laurel essential oils were found to completely inhibit mycelial growth of F. oxysporum at 30.0 and 50.0 µl petri-1 concentrations, respectively. Complete growth inhibition of P. funerea by essential oil of fennel and laurel were observed at relatively lower concentrations (20.0 and 25.0 µl petri-1 concentrations, respectively). Light microscopic observations on hyphae, exposed to volatile phase of the most efficient concentrations of essential oil, revealed considerable structural deformations such as cytoplasmic coagulation, vacuolations and protoplast leakage. In conclusion, our results suggest that essential oils have the potential for use in control of fungal diseases of conifer plants.

Incidence of pine needle blight and its relationship with site factors of Japanese red pine forests in the Kunyushan Mountains, East China

Pine needle blight (Pestalotiopsis funerea (Desm.) Steyaert) has caused major damage to the Japanese red pine (JRP, Pinus densiflora Sieb. et Zucc.) forests in the Kunyushan Mountains region (121°41′34″–121°48′04″ E, 37°11′50″–37°17′22″ N) of Shandong, China. Site factors play an important role in the occurrence of JRP needle blight. Here, the regression equations of the disease incidence (DI) site factors and disease severity index (DSI) site factors were established to predict the incidence and severity of JRP needle blight. In this study, the site factors, DI, and DSI of pine needle blight were surveyed in 121 pure JRP forests plots. The incidence and severity of JRP needle blight were investigated, and regression equations were developed for DI and DSI site factors. The results show that JRP needle blight was a common disease in the Kunyushan Mountains, and 95.9% of the surveyed plots were affected. The DI and DSI of JRP needle blight were significantly affected by altitude, slope angle, and humus depth (p < 0.05), as well as highly significantly affected by slope shape and slope aspect (p < 0.01). The R2 values of DI and DSI models were 0.68 and 0.798, respectively, indicating that their interpretation rate for the occurrence of JRP needle blight is more than 65%, proving that the model is reliable and that key site factors can be used to predict the incidence of JRP needle blight. Overall, the occurrence of pine needle blight in forest land with low altitude, steep and shaded slopes, and thinner humus depth was higher than in other types of sample plots. Thus, the prevention of JRP needle blight should be the focus in this type of woodland.

Comparison of the fungi Pestalotiopsis funerea (Desm.) Steyaert and Truncatella hartigii (Tubeuf) Steyaert isolated from some species of the genus Pinus L. in morphological characteristics of conidia and appendages

The similarity of and differences between the fungi Pestalotiopsis funerea (Desmazières) Steyaert and Truncatella hartigii (Tubeuf) Steyaert that damage the needles of the host plants Pinus ponderosa Peter & Charles Lawson, Pinus coulteri D. Don and Pinus nigra Arnold planted in Mlyňany ArborĂŠtum, were evaluated with emphasis on comparison of the morphology of their conidia and appendages. Conidia of P. funerea were fusiform to ellipsoid, straight to slightly curved, 4-septate with 3 (4-6) apical sometimes branched appendages. The basal cell was short, hyaline, obtuse and thin-walled with basal, hyaline, filiform, straight, unbranched and single appendage. Conidia of T. hartigii were fusiform, 3-septate, without basal appendages. 2-3 apical appendages arising from the apical cell were attenuated, flexuous, sometimes dichotomously branched. The highest occurrence of the fungi P. funerea and T. hartigii indicates the influence of biotic stresses on properties of these pine trees.

Występowanie i szkodliwość Pestalotiopsis funerea dla iglastych roślin ozdobnych

Summary Surveying of coniferous plants in the years 2010–2011 showed the occurrence of necrotic lesions on shoot tips on about 0.5 to 9% of Lawson cypress, juniper, microbiota and thuja. The disease dominated on thuja shoots. Isolates of Pestalotiopsis funerea from juniper and thuja colonised stem parts of different species and cultivars of both plants. Significant differences in susceptibility of juniper and thuja to P. funerea were noticed. Isolates of the species from ericaceous plants caused shoot necrosis of heather and 6 species of coniferous plants. The disease spread, about 2 times faster on heather plants than on juniper and thuja. The trial with P. funerea growth in relation to temperature showed that 20–25°C is an optimal temperature for pathogen development.

Metabolic response of Taxus baccata L. to Pestalotipsis funerea Desm. in dependence of environmental conditions

One of the reasons of yew shoot blight observed in the garden and park plantations was the infection with &lt;i&gt;Pestalotiopsis funerea&lt;/i&gt; fungus. Leaves with extensive disease symptoms have been found to have metabolic disturbances induced by this pathogen. Activation of oxidative stress enzymes, i.e. catalase and peroxidase, points to the possibility of inducing defense responses. Among factors modifying the degree of plant affection under natural conditions, the level of nitric nutrition as well as light conditions may play an important role.

First Report of Botryosphaeria iberica Associated with Dieback and Tree Mortality of Monterey Cypress (Cupressus macrocarpa) in Algeria.

Stem cankers and branches showing bark discoloration, fissuring, resin exudation leading to dieback, crown wilting, and tree mortality have been observed since late spring 2008 on 40-year-old Cupressus macrocarpa (Hartw.) trees planted in forests mixed with Juniperus oxycedrus L. and Acer monspessulanum L. in Taffet, near Ain Abbessa, in the district of Bougaa, Algeria (36°18'57″N; 05°06'33″E; 1,400 m elevation). In 2010, approximately 60% of the C. macrocarpa trees were diseased. For fungal isolations, cankered branches were surface sterilized with ethanol. After removal of the outer bark, fragments of necrotic inner bark taken from the margin of cankers were plated on potato dextrose agar (PDA). Most of the colonies were identified as Botryosphaeria iberica (Phillips, Luque & Alves) based on comparison of morphological traits and DNA sequences with known isolates of the fungus (1). Pestalotiopsis funerea colonies were also obtained, although with less frequency. B. iberica colonies on PDA were dark green with aerial mycelium and optimum growth at 25°C. Pycnidia were produced after 3 weeks of incubation at 20°C under a 12-h near UV light photoperiod on water agar amended with autoclaved cypress seeds. Conidia were brown, one-septate, oval to oblong, and 24.2 (20.1 to 27.4) × 11.2 μm (8.8 to 14.1) (n= 50). An isolate was deposited at the Centralbureau voor Schimmelculture as CBS 130984. DNA was extracted from freeze-dried mycelium and amplified using primers ITS1 and ITS4. The amplified DNA sequence of B. iberica isolate CBS 130984 from Algeria (GenBank Accession No. JN836991) showed 100% homology with sequences of B. iberica isolates obtained from dead and cankered bark of oaks from Spain and Italy (GenBank Accession Nos. AY573216, AY573214, AY573213, AY573210, AY573202, and AY573201). Stem inoculations were performed in the greenhouse on 10 4-year-old, grafted plants of C. macrocarpa growing in 5-liter pots using isolate CBS 130984. A 3-mm plug taken from the margin of a colony grown on PDA for 1 week was inserted in a circular wound of the same size made in the bark with a cork borer where the stem diameter was approximately 1 cm. Inoculations were repeated in June 2010 and June 2011. Five months after inoculations, small rounded to elongated lesions (1.0 to 2.5 cm long), sometimes with resin exuding cracks, were visible on all inoculated stems. Control trees, inoculated with sterile PDA plugs, showed no canker development. B. iberica was successfully reisolated from the necrotic bark surrounding the inoculation sites. No significant differences in canker size were observed between the two replicated experiments. Some Botryosphaeria species that are found on a variety of hosts are also known to cause cankers and dieback of cypress; among these are B. stewensii, B. obtusa, B. dothidea, and B. ribis, often acting as weak pathogens (2,3). Considered weakly virulent in causing dieback of grapevine (4) and, to our knowledge, reported here for the first time on Cupressaceae, B. iberica caused cankers and dieback of C. macrocarpa trees that had probably been weakened by repeated drought events occurring in Algeria during the last 10 years.

Pathogenicity of some fungi isolated from cankers on Cupressus sempervirens var. horizontalis in Turkey

Natural stands of Cupressus sempervirens in Turkey are among the largest forests of this species in the world and are regarded as relicts of the centre of origin of var. horizontalis. In this study, we tested the pathogenicity of some of the most common fungal isolates originating from cankers on C. sempervirens by inoculating the isolates into the inner bark of C. sempervirens seedlings. The internal transcribed spacer (ITS) region of rDNA of the isolates was sequenced and compared with those in the GenBank. Among the isolates, eight ITS taxa were found. The isolates were inoculated into the inner bark of 2 year old C. sempervirens seedlings, on average 90 cm tall and 6 to 12 mm thick at the base. The seedlings were incubated seven weeks in a growth chamber at 70% mean relative humidity and 22.5°C mean temperature. The coaxial length of the lesion around the inoculation point on each seedling was measured. Among the eight ITS taxa Pestalotiopsis funerea, two other species of Pestalotiopsis, and two unidentified species belonging to the class Dothideomycetes caused lesions that were significantly larger than those in the controls while Fusarium sp., Cytospora sp. and an unidentified species belonging to Amphisphaeriaceae did not. In contrast to the Pestalotiopsis species, the two members of Dothideomycetes grew also into the sapwood of the seedlings.

Flammulina Velutipes (Curt.: Fr.) Singer: An Edible Mushroom in Northern Forest of Iran and its Antagonistic Activity Against Selected Plant Pathogenic Fungi

Flammulina velutipes is known as edible and medicinal mushroom. The fungus grow on living treesof Zelkova carpinifolia and dead trees of Diospyros lotus (Stand and fallen) in Behshar forest area. These plantswere found as new hosts for F. velutipes. In vitro antagonistic activity of this mushroom against Bipolarissorokiniana, Ophiostoma ulmi, Pestalotiopsis funerea, Fusarium culmurum, and Fusarium oxysporum wasevaluated on MEA as dual culture. Colonies of the fungus were white farinaceous and powdery on MEA, laterturns yellow. Daily growth of colony was measured 5.36mm on MEA at 25 oC. Growth coefficient was indicated36.64 and 90 mm Petriplate was covered up to 15 days after plating. The strongest and poorest antagonisticindex have been seen against Ophiostoma ulmi and Bipolaris sorokiniana, respectively.

Cultural characteristics and pathogenicity of Pestalotiopsis funerea on Cupressus arizonica

SummaryThe fungal pathogen Pestalotiopsis funerea principally attacks conifer species, causing necrosis on infected tissues, and sometimes death of plants. The main objectives of this study were to determine optimal in vitro growth conditions for Spanish isolates of P. funerea and to evaluate the virulence of these isolates on Cupressus arizonica under field and laboratory conditions, with the goal of verifying possible correlations between these variables. Eight isolates of P. funerea derived from C. sempervirens, C. arizonica and Quercus pyrenaica were used in the assays. In the growth rate experiment, five culture media (PDA, MEA, WA, PCA and TAKAY medium) and six temperatures (5, 10, 15, 20, 25 and 30°C) were evaluated. In the pathogenicity tests, two different experiments were carried out: (i) laboratory inoculations used 30‐mm‐long twigs of C. arizonica; (ii) in field inoculations, twigs of C. arizonica were inoculated with mycelia through wounds. Four months after inoculation, length of the twigs affected by necrosis were measured. The results suggested that Spanish P. funerea isolates had optimum growth at 25°C on TAKAY medium. The fungus caused substantial necroses on C. arizonica twigs after inoculation in both excised twigs and on adult C. arizonica trees. A positive relationship between inoculations carried out under laboratory and field conditions was found. No clear relationships were found between cultural characteristics and pathogenicity for P. funerea.

Spread of plant pathogens and insect vectors at the northern range margin of cypress in Italy

The Mediterranean cypress (Cupressus sempervirens) is a multi-purpose tree widely used in the Mediterranean region. An anthropogenic range expansion of cypress has taken place at the northern margin of the range in Italy in recent decades, driven by ornamental planting in spite of climatic constraints imposed by low winter temperature. The expansion has created new habitats for pathogens and pests, which strongly limit tree survival in the historical (core) part of the range. Based on the enemy release hypothesis, we predicted that damage should be lower in the expansion area. By comparing tree and seed cone damage by pathogens and pests in core and expansion areas of Trentino, a district in the southern Alps, we showed that tree damage was significantly higher in the core area. Seed cones of C. sempervirens are intensively colonized by an aggressive and specific pathogen (the canker fungus Seiridium cardinale, Coelomycetes), associated with seed insect vectors Megastigmus wachtli (Hymenoptera Torymidae) and Orsillus maculatus (Heteroptera Lygaeidae). In contrast, we observed lower tree damage in the expansion area, where a non-aggressive fungus (Pestalotiopsis funerea, Coelomycetes) was more frequently associated with the same insect vectors. Our results indicate that both insect species have a great potential to reach the range margin, representing a continuous threat of the arrival of fungal pathogens to trees planted at extreme sites. Global warming may accelerate this process since both insects and fungi profit from increased temperature. In the future, cypress planted at the range margin may then face similar pest and pathogen threats as in the historical range.

First report of Hakea sericea leaf infection caused by Pestalotiopsis funerea in Portugal

Plant PathologyVolume 53, Issue 4 p. 535-535 Free Access First report of Hakea sericea leaf infection caused by Pestalotiopsis funerea in Portugal M. F. Sousa, M. F. Sousa Centro de Biologia, Departamento de Biologia, Universidade do Minho, Campus de Gualtar, 4710–057 Braga, PortugalSearch for more papers by this authorR. M. Tavares, R. M. Tavares Centro de Biologia, Departamento de Biologia, Universidade do Minho, Campus de Gualtar, 4710–057 Braga, PortugalSearch for more papers by this authorH. Gerós, H. Gerós Centro de Biologia, Departamento de Biologia, Universidade do Minho, Campus de Gualtar, 4710–057 Braga, PortugalSearch for more papers by this authorT. Lino-Neto, T. Lino-Neto Centro de Biologia, Departamento de Biologia, Universidade do Minho, Campus de Gualtar, 4710–057 Braga, Portugal E-mail: tlneto@bio.uminho.ptSearch for more papers by this author M. F. Sousa, M. F. Sousa Centro de Biologia, Departamento de Biologia, Universidade do Minho, Campus de Gualtar, 4710–057 Braga, PortugalSearch for more papers by this authorR. M. Tavares, R. M. Tavares Centro de Biologia, Departamento de Biologia, Universidade do Minho, Campus de Gualtar, 4710–057 Braga, PortugalSearch for more papers by this authorH. Gerós, H. Gerós Centro de Biologia, Departamento de Biologia, Universidade do Minho, Campus de Gualtar, 4710–057 Braga, PortugalSearch for more papers by this authorT. Lino-Neto, T. Lino-Neto Centro de Biologia, Departamento de Biologia, Universidade do Minho, Campus de Gualtar, 4710–057 Braga, Portugal E-mail: tlneto@bio.uminho.ptSearch for more papers by this author First published: 24 August 2004 https://doi.org/10.1111/j.1365-3059.2004.01042.xCitations: 5 AboutSectionsPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Hakea sericea (Proteaceae) is native to south-eastern Australia and has been considered as an invader of natural habitats. In northern Portugal, dense stands are rapidly spreading after forest fires. In May 2003, unusual leaf spots were observed on naturally growing plants. Infected plants exhibit reddish leaves bearing black, 1–3 mm circular lesions. Leaf sections containing necrotic lesions were plated onto potato dextrose agar (PDA) and eight fungal isolates were obtained. Pure cultures exhibited pinkish mycelium, bearing compact acervuli that contained black slimy spore masses. Microscopic observation revealed typical Pestalotiopsis sp. five-celled spores (three coloured median and two hyaline end cells) with three to four apical and one basal appendages. Genomic DNA from fungal isolates was purified using a DNeasy® Plant Mini Kit (Qiagen, Crawley, UK), and employed in thermocyclic amplifications using Ready-To-Go PCR beads (Amersham Biosciences, Barcelona, Spain) with ITS5 and ITS4 internal transcribed spacer (ITS) universal primers (White et al., 1990). The amplified sequences (599 bp) were analysed together with other Pestalotiopsis sequences (Jeewon et al., 2002) using the programs ClustalX, GeneDoc and Phylip. ITS sequences from all fungal isolates were identical to each other and only four base pairs were different (99·3% similarity) from Pestalotiopsis funerea. To confirm the pathogenicity of P. funerea on H. sericea, leaf-wounded and nonwounded 6-week-old in vitro plants were sprayed with a suspension containing 105 spores mL−1 or with sterile water. After 6 days, only leaf-wounded plants infected by P. funerea exhibited lesions identical to those observed in field plants, and bearing fungal spores identical to those from original isolates. Previously, species of Pestalotiopsis have been described as pathogenic on Proteaceae species (Taylor et al., 2001). In countries where H. sericea invasion has become a problem, its biological control has been successfully achieved using Colletotrichum gloeosporioides (Richardson & Manders, 1985). The identification of naturally occurring pathogenic fungi in Portuguese H. sericea may allow the design of an integrated control strategy for this invading species. References Jeewon R, Liew ECY, Hyde KD, 2002. Phylogenetic relationships of Pestalotiopsis allied genera inferred from ribosomal DNA sequences and morphological characters. Molecular Phylogenetics and Evolution 25, 378– 92.CrossrefCASPubMedWeb of Science®Google Scholar Richardson DM, Manders PT, 1985. Predicting pathogen-induced mortality in Hakea sericea (Proteaceae), an aggressive alien plant invader in South Africa. Annals of Applied Biology 106, 243– 54. Wiley Online LibraryWeb of Science®Google Scholar Taylor JE, Crous PW, Palm ME, 2001. Foliar and stem fungal pathogens of Proteaceae in Hawaii. Mycotaxon 78, 449– 90. Web of Science®Google Scholar White TJ, Bruns T, Lee S, Taylor J, 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: MA Innis, DH Gelfand, JJ Sninsky, TJ White, eds. PCR Protocols – A Guide to Methods and Applications. San Diego, CA, USA: Academic Press, 315– 22. Google Scholar Citing Literature Volume53, Issue4August 2004Pages 535-535 ReferencesRelatedInformation

First Report of Pestalotiopsis funerea on Cupressocyparis leylandii in Italy.

Pestalotiopsis funerea (Desm.) Steyaert occurs worldwide and is reported as an opportunistic invader of conifers. It can also be have as a primary pathogen. During the spring of 2001, leaf yellowing was observed on mature and young Cupressocyparis leylandii trees in some nurseries in northern Italy. Symptoms were first observed on twig tips and progressed toward the stem during the summer, resulting in twig dieback and death of branches up to 15 mm. Dark acervuli were present in the cankers. Isolations were made from cankers and necrotic twigs by removing the outer tissue and placing wood slivers from affected tissues on potato dextrose agar. P. funerea was isolated from 88 and 72% of cankers and necrotic twigs, respectively. Four isolates were selected for inoculation tests. Inoculations were made in September on the upper part of stems of 4-year-old pot-grown C. leylandii saplings. The outer layer of the bark (5 × 3 mm) was removed, and a 5-mm-diameter mycelial plug on malt extract agar (MEA) was placed on the wound and covered with Parafilm. Sterile MEA plugs served as controls. The experiment included 20 replications for each isolate and the control. After 4 weeks, 80% of inoculated plants showed bark necrosis or small cankers, or both, and 20% died. P. funerea was reisolated. No cankers developed in the controls. P. funerea was previously reported in Italy on Thuja orientalis L. (2) and from France on C. leylandii (1), but to our knowledge, this is the first report of the pathogen on C. leylandii in Italy.

Susceptibility of cones and seeds to fungal infection in a pine (Pinus spp.) collection

Summary Thirty‐one fungi, members of Ascomycota and Deuteromycota, were isolated from cones/seeds of 28 hosts (Pinus spp.) originating from East Asia, Europe and North America, and growing at the Montreal Botanical Garden, Canada. A total of 21 taxa of these isolated fungi are considered pathogens. The damage severity was most prevalent on Diploxylon pines of European provenance (Pinus nigra, Pinus mugo and Pinus sylvestris), and one indigenous North American species (Pinus albicaulis). Twelve fungi occurred on different hosts, i.e. Sphaeropsis sapinea (12), Herpotrichia juniperi (8), Truncatella hartigii (7), Ceuthospora sp. (5), Fusarium, spp. (5), Pestalotiopsis funerea (3), Phomopsis sp. (3), Valsa spp. (anamorph. Cytospora) (3), Diaporthe sp. (3), Fusicoccum sp. (2) and Sirococcus strobilinus (2). Sphaeropsis sapinea, Herpotrichia juniperi, Phomopsis conorum, Truncatella hartigii, Tubercularia sp. and Valsa spp. were related to high cone and seed damage. For the most frequently observed Sphaeropsis sapinea damage, the subgenus Haploxylon was less susceptible than Diploxylon, and P. sylvestris less than P. nigra and P. mugo. Pinus resinosa showed more tolerance to S. sapinea. Truncatella hartigii was more frequently associated with necrotic lesions on cones/seeds in comparison with Pestalotiopsis funerea. The results are discussed in the context of arboretum, seed orchard, nursery and quarantine management.

Efficient transmission of an introduced pathogen via an ancient insect-fungus association

In Cupressus sempervirens the association between seed insects and tree pathogens has resulted in optimal exploitation of the cones. A fungus-infected cone can be inhabited by the nymphs of a true seed bug (Orsillus maculatus), the adults of which may carry a heavy spore load at emergence. Cones are infected when eggs are laid within the cone, most frequently via the emergence holes of a seed wasp (Megastigmus wachtli). This symbiotic association evolved with the nonaggressive fungus Pestalotiopsis funerea within the natural range of the cypress. When the aggressive cypress canker disease (Seiridium cardinale) was introduced into Europe, it was transmitted by O. maculatus to cones usually colonized by Pestalotiopsis funerea, with disastrous consequences for the regeneration and survival of C. sempervirens in the entire Mediterranean area.http://link.springer. de/link/service/journals/00114/bibs/9086010/90860479.htm</HEA

The ecology of fungal cankers on Cupressus macrocarpa in southern England

SummaryBark isolations from cankers on Cupressus macrocarpa in southern England yielded Seiridium cardinale, Pestalotiopsis funerea and Pestalotiopsis monochaetioides. The recovery pattern of the three fungi indicated that only S. cardinale was a primary pathogen; the two Pestalotiopsis species being secondary. Pathogenicity data obtained following the inoculation of C. macrocarpa branches with mycelium of the three fungi confirmed this view.No evidence of vegetative incompatibility was found in S. cardinale, but five vegetative‐compatibility (vc) groups were found in P. funerea and six in P. monochaetioides. Different vc groups of the two Pestalatiopsis species were often found in the same S. cardinale lesion, suggesting that a number of separate colonization events were involved. It is possible that this sometimes leads to complete replacement of the pathogen.