Incidence and severity of dieback in birch plantings associated with Anisogramma virgultorum and Marssonina betulae in Scotland

Betula pendula and B. pubescens were surveyed at nine Scottish Woodland Grant Scheme (WGS) sites in 2004 to assess the incidence and severity of two fungal pathogens, Anisogramma virgultorum and Marssonina betulae , and their association with crown dieback. Of the surveyed trees, 47% had at least 40% crown dieback, with B. pendula more severely affected than B. pubescens at most sites. Overall, 57% of surveyed birch trees had A. virgultorum and 28% had M. betulae , with the incidence of trees infected with both species varying among sites. Incidence and severity of A. virgultorum were greater on B. pubescens than on B. pendula , whereas the reverse was true for M. betulae. The relationship between incidence of M. betulae foliar disease and sunken cankers was significant, with 83% of M. betulae -infected trees having these cankers. The presence of M. betulae foliar disease resulted in a greater percentage of crown dieback compared with non-infected trees in both birch species, whereas the presence of A. virgultorum caused greater crown dieback in B. pubescens only. Across all sites and both birch species, as severity of infection with either A. virgultorum or sunken cankers increased, so did the severity of crown dieback. This study showed that A. virgultorum and M. betulae appear to contribute significantly to birch dieback at the nine sites surveyed.     

Phomopsis alnea, the cause of dieback of black alder in Italy

Black alder ( Alnus glutinosa ) occurs naturally on moist soils and along streams and rivers throughout most of its range. Groves of this species in north-central Italy were recently found to be attacked by the mitosporic coelomycete Phomopsis alnea , which causes perennial stem cankers and dieback. The fungi Melanconium apiocarpium and a Hymenopsis sp. were also frequently found on necrotic alder tissue, occasionally invading the living bark. All these fungi were tested for pathogenicity in two seasonal inoculation trials on seedlings of black alder, Italian alder ( Alnus cordata ) and green alder ( Alnus viridis ) that were either normally watered or water-stressed. Phomopsis alnea actively colonized the seedlings and reproduced the symptoms observed in the field. The other fungi behaved as weak parasites, only occasionally spreading to apparently healthy bark tissue. It appears that these fungi are saprobes, commonly colonizing bark and twigs already killed by P. alnea . Symptoms caused by P. alnea in the field were exacerbated on dry sites and by seasonal drought stress. On artificially inoculated and water-stressed seedlings, both the incidence and severity of P. alnea also increased, causing extensive mortality. The data provide evidence for the belief that P. alnea becomes a factor in the dieback of natural black alder woodlands when trees are first impaired by wounding agents and then subjected to unusual extended drought.     

Anastomosis group and pathogenicity of isolates of Rhizoctonia solani from potato crops in South Australia

Isolates of Rhizoctonia collected from the stems, roots, tuber sclerotia and soil of potato crops in Virginia and Lenswood, South Australia, were identified to anastomosis groups (AG). Of the 301 multinucleate isolates of Rhizoctonia solani tested, 90% were AG-3, 7% were AG-4 and 2% were AG-5; 12 isolates were binucleate Rhizoctonia spp. This is the first report of isolates of AG-4 and AG-5 causing disease in potato crops in South Australia. All AG-3, AG-4 and AG-5 isolates tested caused rhizoctonia disease symptoms on the potato cultivar Coliban in pathogenicity trials conducted under glasshotise conditions. Both AG-3 and AG-5 isolates caused black scurf and stem cankers, although symptoms of black scurf were less severe with AG-5. AG-4 isolates produced the most severe stem and stolon cankers of all isolates tested. The pathogenicity of tuber-borne inoculum was confirmed by growing plants from sclerotia-infested tubers. AG-8 isolates from diseased barley and wheat produced severe root cankers and caused loss of feeder roots on inoculated potato plants. Results suggest that rhizoctonia disease in potato fields in South Australia is caused by a combination of different anastomosis groups and this has important implications for crop rotations.     

Phytophthora ramorum and Phytophthora kernoviae on naturally infected asymptomatic foliage

Phytophthora ramorum and Phytophthora kernoviae are recently discovered invasive Phytophthoras causing leaf necrosis and shoot tip dieback mostly on ornamental and forest understorey species, but also cause bleeding cankers on stems of a wide range of tree species. Sporulation occurs only on infected shoots or fruits and foliage so foliar hosts are central to the disease epidemiology. In field trials to assess infection in trap plants exposed to natural inoculum of P. ramorum and P. kernoviae on rhododendron in south west England, it was discovered that leaves of the trap plants ( Rhododendron 'Cunninghams White') and holm oak ( Quercus ilex ) were asymptomatically infected and supported sporulation of both pathogens. More than half the rhododendron trap plants exposed to inoculum of P. kernoviae became infected compared with approximately a third of those exposed to P. ramorum in a natural situation. Approximately one third of the infections were detected from asymptomatic foliage for both pathogens. The significance of these findings for plant health regulation based on visual inspection as a measure to prevent introduction and dissemination of both these pathogens is explored and research gaps identified.     

Pathogenicity and genetic variation of Phytophthora cactorum from silver birch and strawberry

Phytophthora cactorum strains isolated from necrotic stem lesions on Betula pendula seedlings or from Fragaria ananassa plants suffering from crown rot were pathogenic to their host plants. Only isolates from birch caused clear lesions on non-wounded bark of birch. P. cactorum isolates from birch were not detrimental to strawberry. Random Amplified Polymorphic DNA (RAPD) analysis revealed variation within P. cactorum, isolates from silver birch having different banding patterns than those from strawberry. UPGMA analysis clustered isolates from silver birch and strawberry plants into separate groups. The data show that the recent outbreak in Finland of P. cactorum in birch could not be caused by the import of strawberry plants affected by crown rot.     

Colletotrichum gloeosporioides as the cause of stem tip dieback of cassava

A fungus with morphological features corresponding to the group species Colletorichum gloeosporioides was consistently isolated from cassava with shoots showing dieback symptoms in Ghana. When four locally-grown cultivars were inoculated with isolates of the fungus, they developed disease symptoms, which consisted of discrete dark brown lesions on the stems followed by defoliation. Koch's postulates were completed by re-isolating the fungus from the inoculated plants. The relatedness of the isolates lo other members of the genus Calletotrichum , whose identities were well established, was investigated by comparison of the nucleotide sequence of domain 2 of their ribosomal DNA. The cassava isolates differed from authentic isolates of C. gloeosporioides by only one nucleotide among the 193 analysed. The causal agent of cassava stem tip dieback (STDB) is thus identified as a form of C. gloeosporioides.     

Characterization of Cytospora isolates from wood cankers of declining grapevine in North America,with the descriptions of two new Cytospora species

Cytospora species are ubiquitous pathogens of numerous woody plants, causing dieback and wood cankers in agronomic crops, timber trees and wildland trees (e.g. Prunus, Eucalyptus and Salix, respectively). Cytospora chrysosperma, C. cincta and C. leucostoma have been reported from grapevines in Iran showing symptoms of one or more recognized trunk diseases (esca, botryosphaeria‐, eutypa‐ and phomopsis diebacks); however, only C. chrysosperma was shown to be pathogenic to grapevine. To understand the potential role of Cytospora species in the grapevine trunk‐disease complex, 21 Cytospora isolates were examined that were recovered from dieback and wood cankers of Vitis vinifera and Vitis interspecific hybrids in seven northeastern U.S. states and two Canadian provinces. Phylogenetic analyses of ITS and translation elongation factor 1‐α identified two novel species: Cytospora vinacea sp. nov. and Cytospora viticola sp. nov. Differences in culture morphology and conidial dimensions also distinguished the species. When inoculated to the woody stems of potted V. vinifera ‘Thompson Seedless’ in the greenhouse, both species were pathogenic, based on development of wood lesions and fulfilment of Koch's postulates. Cytospora viticola was the most virulent based on lesion length at 12 months post‐inoculation. As cytospora canker shares some of the same general dieback‐type symptoms as botryosphaeria‐, eutypa‐ and phomopsis diebacks, it may be considered part of the grapevine trunk‐disease complex in eastern North America.     

Pathogenicity of fungi associated with winter loss and injury in white lupin

Fungi isolated from diseased white lupin plants were tested for pathogenicity in standardized glasshouse tests to assess their potential as causes of plant death in winter in autumn-sown crops. Pleiochaeta setosa and, to a slightly lesser extent, Fusarium avenaceum caused lesions or plant death when inoculated onto hypocotyls of lupin seedlings. Disease was more severe when the hypocotyls were wounded before inoculation. F. avenaceum also caused injury when inoculated onto roots; wounding the roots had no effect. Some isolates of F. solani also caused injury, especially after root inoculation, but isolates of F. oxysporum and Cylindrocarpon destructans were only slightly or non-pathogenic. Isolates of Fusarium and C. destructans , but not of P. setosa , differed in pathogenicity. There were differences in susceptibility to disease among three lupin cultivars.     

The effect of Cryphonectria parasitica on water relations of American chestnut

Chestnut blight has commonly been regarded as a phloem disease due to conspicuous stem cankers that result from infection by the fungal pathogen Cryphonectria (Endothia) parasitica. Stomatal conductance (g_s), transpiration rate (E) and leaf water potential were measured throughout the day on leaves distal to naturally-occurring virulent (sunken bark with abundant stromata) or hypovirulent (swollen bark lacking stromata) cankers and cankers induced by inoculation with virulent or hypovirulent strains of C. parasitica. Relative to control stems, hydraulic conductivity (K_h), g_s and E were reduced significantly (α = 0·05) for both natural virulent cankers, and cankers that were induced by a virulent strain. These effects were less pronounced for both natural and induced hypovirulent cankers. Isolation experiments revealed that the percentage of xylem tissue chips yielding C. parasitica was greater for virulent than for hypovirulent cankers. The data provide evidence that the localized presence of C. parasitica in cankers of American chestnut results in stomatal closure, possibly as a direct result of xylem dysfunction.     

Identification,potential inoculum sources and pathogenicity of botryosphaeriaceous species associated with grapevine dieback disease in New Zealand

This study investigated the prevalence and identity of botryosphaeriaceous dieback pathogens in necrotic grapevines tissues in New Zealand vineyards, and other woody hosts growing nearby. The presumptive identities of the isolates by conidial and cultural morphology were confirmed with ITS sequence data as Neofusicoccum australe, N. luteum, N. parvum and Diplodia seriata. They were isolated predominantly from necrotic stems of grapevine and other hosts, but also from leaves, flowers and wood debris of grapevines. Inoculation with conidia and mycelium of multiple isolates of each species onto excised and attached green shoots and trunks of five grapevine varieties, Cabernet sauvignon, Chardonnay, Pinot noir, Riesling, and Sauvignon blanc, showed that all varieties became infected to a similar extent. All species except D. seriata were pathogenic, irrespective of the host source, with N. luteum being the most and D. mutila the least pathogenic (P < 0.05). On trunks, N. parvum caused cankers and the other pathogenic species caused die-back when the inoculated vines became winter-dormant. Conidia were produced from green shoot lesions and die-back wood, which indicates potential inoculum sources for vineyard infection.     

Pestalotiopsis canker of cypress in Israel

The occurrence of a new type of canker on stems and branches of cypress trees(Cupressus sempervirens L.) in Israel is reported.Pestalotiopsis funerea (Desm.) Steyaert was isolated from the cankers. Its pathogenicity was confirmed by formation of typical disease symptoms in inoculated plants and reisolation of that fungus from the cankers. Canker development was significantly enhanced when extreme water stress (water potential of -4.7 MPa) was imposed prior to inoculation.     

Evaluation of fungicides for the management of Botryosphaeria dieback diseases of grapevines

BACKGROUND: A range of botryosphaeriaceous species can cause dieback and cankers in grapevines; however, different species most commonly affect the grapevines in different grape‐growing regions and countries. They infect through wounds and sporulate on woody stems and green shoots throughout the year, so wound protection is the recommended control strategy. This research evaluated fungicides for their ability to reduce mycelial growth and conidial germination of three botryosphaeriaceous species and to protect pruning wounds against infection. RESULTS: In vitro experiments showed that nine out of 16 tested fungicides were effective at reducing mycelial growth and/or conidial germination of three isolates each of Neofusicoccum australe, N. luteum and Diplodia mutila. The species differed in their response to the fungicides, although N. luteum was usually the least sensitive. When nine selected fungicides were sprayed on cane pruning wounds on potted and field grapevines and subsequently inoculated with N. luteum conidia, some effectively protected them from infection. The most effective fungicides were flusilazole, carbendazim, tebuconazole, thiophanate‐methyl and mancozeb, as they prevented the inoculated pathogen from infecting healthy wood in 100, 93, 87, 83 and 80% of field vines, respectively. CONCLUSION: This research has demonstrated that fungicides applied after winter pruning can protect vines from infection by conidia of three botryosphaeriaceous species. Copyright © 2012 Society of Chemical Industry     

松材线虫对黑松一些生理生化指标的影响（英文）

 松材线虫病是松树的一种毁灭性病害。本文对接种松材线虫后3~4年生黑松茎部的一些生理生化指标的变化进行了研究。结果显示,接种线虫后,松苗的可溶性糖含量降低,在病情发展中期和后期比早期降低更显著。松苗的可溶性蛋白质和抗坏血酸含量逐渐降低,游离氨基酸含量也降低。松树茎部的可溶性蛋白质含量可用于病害的早期诊断。此外,对这4个生理生化指标在病害致病机理中的作用进行了讨论。     

Phytophthora kernoviae and P. ramorum: host susceptibility and sporulation potential on foliage of susceptible trees1

Phytophthora kernoviae and P. ramorum are introduced, invasive pathogens in the UK. Both species are adapted for aerial dispersion and have a wide host range, many of which are common to both pathogens. The diseases they cause are foliar necrosis and shoot tip dieback on both tree and ornamental hosts, and bleeding cankers on tree hosts. Inoculum is produced on infected foliage but not on bleeding cankers in both cases. Proactive measures to prevent disease spread and to evaluate the risks posed by these pathogens are being undertaken. Amongst others, these include using the detached leaf assay to get an indication of tree foliage susceptibility, and inoculating wounded stems of saplings to get an idea of under‐canopy sapling and nursery tree susceptibility. The sporulation potential on selected susceptible hosts was assessed, and finally, surveys which are still ongoing were carried out in south‐west England. Results of this work are presented and discussed.     

Systemic growth of Leptosphaeria maculans from cotyledons to hypocotyls in oilseed rape: influence of number of infection sites, competitive growth and host polygenic resistance

The influence of competitive effects between two isolates, of the number of infection sites on cotyledons and of host polygenic resistance on the systemic growth of Leptosphaeria maculans , the cause of phoma stem canker in oilseed rape ( Brassica napus ), were investigated. Controlled-condition experiments were conducted with two oilseed rape doubled haploid lines, one susceptible and the other with a high level of polygenic resistance, inoculated via wounded cotyledons with conidial suspensions obtained from two isolates. Expression of cankers in plants was enhanced by exposing inoculated plants to low temperature (6°C) followed by warm temperature (20°C). The fungus was detected by PCR amplifications of three minisatellite markers in all stems with visible canker symptoms and also in the stems of 14 of the 59 plants without visible cankers on the hypocotyls. Disease severity increased with the number of infection sites on cotyledons: in one of the three replicate experiments, the mean external necrosis length on the hypocotyl ranged from 6·47 to 35·3 mm for one and eight infections sites on cotyledons, respectively. The probability of an isolate reaching the hypocotyl from inoculated cotyledons decreased with increasing competing inoculum load on cotyledons: for instance, for isolate A290v it decreased from 1 when inoculated alone to 0·28 when coinoculated with six drops of competing isolate P27d. Polygenic resistance significantly reduced disease incidence and severity. For instance, in one of the three replicate experiments, disease incidence ranged from more than 74% in susceptible plants to 16% in resistant ones, while mean external necrosis length was up to 35·3 and 6·5 mm on susceptible and on resistant plants, respectively. This study offers new possibilities for assessing levels of polygenic resistance to stem canker in B. napus and studying the aggressiveness of L. maculans isolates.     

Fungi as potential factors limiting natural regeneration of pedunculate oak (Quercus robur) in mixed-species forest stands in Poland

Dieback of young Quercus robur seedlings can limit natural regeneration in mixed-species forest stands in Poland. The aim of this study was to examine the role of fungi in the dieback of oak seedlings in central Europe. Fungi were isolated from the stems and roots of Q. robur, from both healthy seedlings and seedlings with symptoms, that were sampled from four stands in Poland. In total, 111 distinct taxa were identified. Ascomycota was dominant, representing 95.6% of the isolates. Among the taxa identified, Alternaria alternata, Colletotrichum fioriniae, Colletotrichum godetiae, Coniella quercicola, Diaporthe eres, Gnomoniopsis paraclavulata, Ilyonectria rufa, Mucor genevensis, Penicillium glandicola, Tubakia dryina, and Umbelopsis changbaiensis were most frequently isolated, and were consistently found in stems and roots of both healthy and diseased plants. The community compositions of fungi in healthy and diseased stems were similar, although Fusarium species, especially Fusarium sp. 1 (FTSC 5) and F. sporotrichioides, were found mainly in seedlings with symptoms. The pathogenicity of the most consistently isolated species from stems and roots of diseased seedlings was tested on Q. robur seedlings. F. sporotrichioides caused the largest lesions on inoculated seedlings. Six weeks after inoculation, D. eres, C. fioriniae, G. paraclavulata, T. dryina, and F. sporotrichioides killed 0%–18.8% of seedlings, while I. rufa and Ilyonectria pseudodestructans did not cause any lesions or other symptoms. This study is the first comprehensive report suggesting that massive fungal attack can lead to oak seedling dieback in mixed-species forest stands in central Europe.     

Variation in host range, systemic infection and epidemiology of Leptosphaeria maculans

The A and B groups (aggressive and non-aggressive) of Leptosphaeria maculans were compared in studies of host range, infection phenotypes and epidemiology. Isolates of both groups infected a wide range of cruciferous hosts including Brassica napus, B. rapa, B. oleracea, B. juncea, B. carinata. B. nigra, Thlaspi arvense and Raphanus sativus. On cotyledons, B-group isolates were generally more aggressive than A-group isolates, causing local lesions and subsequent systemic invasion of the majority of test species. On susceptible stems, A-group isolates caused cortical lesions; B-group isolates, if they reached the stem, usually caused pith lesions with no external symptoms. In a susceptible line of B. napus , CrGC5, systemic infections of the leaf and petiole were similar with both groups, but the B-group rarely entered the stem to form cortical lesions or cankers. At two different field locations, natural epidemics caused by the two groups on oilseed rape leaves were similar in overall pattern, B-group lesions occurred slightly later but the incidence increased more rapidly and reached a maximum slightly earlier than that of the A group. The two groups differed markedly in stem infection patterns. Infection of the cortex near the base of the stem and the development of a typical stem canker was caused entirely by the A group. However the B group often caused considerable damage to the pith. Superficial chlorotic lesions on stems and inflorescences were mainly attributable to the B group. In view of the stem pith infection by the B group without external symptoms, its importance on oilseed rape may have been underestimated previously.     

Phenotypic,molecular and pathogenic characterization of Phlyctema vagabunda,causal agent of olive leprosy

Olive leprosy, caused by the fungus Phlyctema vagabunda, is a classic fruit rot disease widespread in the Mediterranean basin. From 2009 to 2013, new disease symptoms consisting of small circular necrotic leaf lesions, coin branch canker and shoot dieback were observed in Spanish and Portuguese olive orchards showing intense defoliation. Phlyctema‐like anamorphs were consistently isolated from leaves and shoots with symptoms. Representative isolates from affected leaves, shoots and fruits were characterized based on morphology of colonies and conidia, optimum growth temperature and comparison of DNA sequence data from four regions: ITS, tub2, MIT and rpb2. In addition, pathogenicity tests were performed on apple and olive fruits, and on branches and leaves of olive trees. Maximum mycelial growth rate ranged between 0.54 and 0.73 mm per day. Conidia produced on inoculated apple fruits showed slight differences in morphology among the representative fungal isolates evaluated. Phylogenetic analysis clustered all of the Phlyctema‐like isolates in the same clade, identifying them as Phlyctema vagabunda. On fruits, influence of wounding, ripening and cultivar resistance was studied, with cv. Blanqueta being the most susceptible cultivar. On branches, a mycelial‐plug inoculation method reproduced olive leprosy symptoms and caused shoot dieback. On leaves, Koch's postulates were fulfilled and the pathogen caused characteristic necrotic spots and plant defoliation. This is the first time that the pathogenicity of P. vagabunda in olive leaves has been demonstrated.     

Aetiology of branch dieback,panicle and shoot blight of pistachio associated with fungal trunk pathogens in southern Spain

Pistachio represents an emerging nut crop across the Mediterranean basin. In Spain, pistachio has been traditionally cultivated in marginal-dry areas with unfavourable climatic conditions for plant diseases. Consequently, little attention has been given to research on pistachio diseases until recently. Symptoms of branch dieback and cankers, and shoot and panicle blight have been recently observed in commercial pistachio orchards across southern Spain. In this study, 10 commercial pistachio orchards showing disease symptoms were surveyed between 2017 and 2018. Botryosphaeriaceae fungi were consistently isolated from affected shoots, among other fungal families with minor relevance. Representative isolates of each family were characterized based on colony and conidial morphology, optimum growth temperature, and the comparison of DNA sequence data (ITS, LSU, EF, TUB2, and ACT genomic regions). Detached and attached shoots, and attached panicles of pistachio cv. Kerman were inoculated with mycelial plugs or conidial suspensions to demonstrate the pathogenicity of the selected isolates. Botryosphaeria dothidea, Lasiodiplodia pseudotheobromae, Neofusicoccum mediterraneum, N. parvum, Diaporthe neotheicola, Diaporthe sp., Eutypa lata, Eutypa sp., Cytospora sp., and Phaeoacremonium minimum were identified. P. minimum had the highest optimum growth temperature (29.6 °C) and Cytospora sp. the lowest (21–22 °C). Botryosphaeriaceae isolates showed the largest lesions on inoculated shoots, with L. pseudotheobromae being the most aggressive, followed by Neofusicoccum species. Panicles inoculated with N. mediterraneum showed blight symptoms and canker formation 6 weeks after inoculation, without significant differences in aggressiveness between isolates. This work reports relevant information about this emerging disease in the novel Spanish pistachio-growing areas.     

Characterization of Neofusicoccum parvum causing canker and dieback on Brachychiton species

Brachychiton species are planted in Italy as ornamental trees. A survey in a nursery in Sicily (Italy) revealed the presence of young B. acerifolius and B. populneus showing severe trunk cankers, massive gummosis from the bark cracks, wood discolouration, and twig dieback. Morphological characterization was based on conidia measurement, and molecular characterization was carried out performing phylogenetic analysis (Maximum Parsimony) based on multi-locus approach of partial ITS, EF-1α, and tub2. Identification of the causal agent resulted in Neofusicoccum parvum (Botryosphaeriaceae). Pathogenicity tests were conducted in order to fulfill the Koch’s postulates. Five and three-years-old potted Brachychiton plants were inoculated (indoor and outdoor) using mycelial plugs. All the inoculated plants showed severe symptoms similar of those observed in the nursery during the survey. Some of the inoculated plants were completely dead. Re-isolations fully confirmed the causal agent. For our knowledge, this is the first report worldwide of N. parvum attacking Brachychiton spp.