Involvement of Phytophthora spp. in chestnut decline in the Black Sea region of Turkey

Chestnut blight caused by Cryphonectria parasitica is a serious disease of Castanea sativa in the Black Sea region of Turkey. During disease surveys, dieback and decline symptoms were observed on trees without apparent blight and ink disease symptoms. Black necroses, similar to those caused by Phytophthora infections, were noted on some of the chestnut coppices and saplings in one nursery in Ordu and led to an investigation into this disease complex. Only symptomatic plants showing dieback symptoms were investigated. Soil samples together with fine roots were collected from two directions, north and north‐east, approximately 150 cm away from the main stems. Phytophthora spp. were baited with young chestnut leaves. Three Phytophthora spp., P. cambivora, P. cinnamomi and P. plurivora, were identified from 12 soil samples collected from 73 locations, while from the nurseries, only P. cinnamomi was obtained. Phytophthora cinnamomi was the most common species, obtained from seven locations in five provinces and from four nurseries having similar symptoms mentioned above in different locations. Phytophthora cambivora and P. plurivora were less frequently obtained, from three to two stands, respectively. Phytophthora cinnamomi and P. cambivora were the most aggressive species when inoculated at the stem base on 3‐year‐old chestnut saplings, killing six saplings of eight inoculated in 2 months. The three Phytophthora species were first recorded on chestnut in Black sea region of Turkey with the limited samples investigated in a large area about 150 000 ha chestnut forest.     

Phytophthora species associated with dieback of sweet chestnut in Western Turkey

Sweet chestnut (Castanea sativa) is an important tree species in the Marmara and Aegean regions of Turkey as these two regions produce the great majority of edible nuts, especially those used for marron glacé production. Chestnut forests and orchards in these regions showing severe dieback symptoms not associated with chestnut blight were investigated to determine the role of Phytophthora spp. in the decline syndrome. Soil samples were collected from around 108 symptomatic chestnut trees at 29 sites and Phytophthora spp. isolated using soil baiting technique and selective medium. Species isolated were identified by cultural characteristics and ITS sequencing. Phytophthora x cambivora was the dominant species detected in 13 sites, followed by P. cinnamomi (5 sites), P. plurivora (3 sites) and P. cryptogea (1 site). Phytophthora x cambivora was present in both regions, while P. cinnamomi was found only in the Marmara region in coastal areas around Istanbul. When inoculated at the stem bases of 3‐year‐old chestnut saplings, P. cinnamomi produced significantly longer necrotic lesions (7.8–12.0 cm) than P. x cambivora (2.6–6.3 cm) by 12 days after inoculation. Phytophthora plurivora was the least aggressive species causing only small lesions. Phytophthora cryptogea, which represents the first record on chestnut in Turkey, produced intermediate sized lesions in between P. x cambivora and P. plurivora. These results indicate that P. x cambivora and in some areas P. cinnamomi play major roles in the observed dieback of sweet chestnut in western Turkey.     

三种橡胶树乙烯利刺激剂对柑桔褐腐疫霉的毒力测定

通过含毒介质和橡胶叶片接种的试验方法,测定三种不同载体的乙烯利涂剂(含2％乙烯利)对柑桔褐腐疫霉的抑菌率,结果表明三种乙烯利涂剂均有不同程度的抑菌能力,其中抗病增胶灵抑菌能力最高,效能稳定,持效期长;而以淀粉和甲基纤维素为载体的涂剂抑菌能力较低,效能不稳定,持效期短。     

Triazole induced drought tolerance in horse chestnut (Aesculus hippocastanum)

We determined the influence of the triazole derivatives paclobutrazol, penconazole, epixiconazole, propiconazole and myclobutanil on the drought tolerance and post drought recovery of container-grown horse chestnut (Aesculus hippocastanum L.) saplings. Myclobutanil neither conferred drought resistance, as assessed by its effects on a number of physiological and biochemical parameters, nor affected growth parameters measured after recovery from drought. Chlorophyll fluorescence (F(v)/F(m)), photosynthetic rates, total foliar chlorophyll and carotenoid concentrations, foliar proline concentration and superoxide dismutase and catalase activities were consistently higher and leaf necrosis and cellular electrolyte leakage was lower at the end of a 3-week drought in trees treated with paclobutrazol, penconazole, epixiconazole or propiconazole than in control trees. Twelve weeks after drought treatment, leaf area and shoot, root and total plant dry masses were greater in triazole-treated trees than in control trees with the exception of those treated with myclobutanil. In a separate study, trees were subjected to a 2-week drought and then sprayed with paclobutrazol, penconazole, epixiconazole, propiconazole or myclobutanil. Chlorophyll fluorescence, photosynthetic rate, foliar chlorophyll concentration and catalase activity over the following 12 weeks were 20 to 50% higher in triazole-treated trees than in control trees. At the end of the 12-week recovery period, leaf area and shoot, root and total plant dry masses were higher in triazole-treated trees than in control trees, with the exception of trees treated with myclobutanil. Application of triazole derivatives, with the exception of myclobutanil, enhanced tolerance to prolonged drought and, when applied after a 2-week drought, hastened recovery from drought. The magnitude of treatment effects was in the order epixiconazole approximately propiconazole > penconazole > paclobutrazol > myclobutanil.     

Parasitic Ichneumonoidea on the horse chestnut leaf miner, Cameraria ohridella (Lepidoptera: Gracillariidae) in Slovakia

Parasitic Hymenoptera from the suprafamily Ichneumonoidea attacking the horse chestnut leafminer Cameraria ohridella Deschka & Dimic (Lepidoptera: Gracillariidae) were studied in southwest Slovakia during 2000 and 2001. The search for such species resulted in the determination of eight ichneumonid species, Gelis agilis (Fabricius), Itoplectis alternans (Gravenhorst), I. clavicornis (Thompson), I. maculator (Fabricius), Scambus annulatus (Kiss), S. brevicornis (Gravenhorst), Zatypota percontatoria (Müller), Adelognathus sp. and a braconid Colastes braconius Haliday. The most abundant species was S. annulatus. All these species recorded are polyphagous and often facultative hyperparasitoids, except C. braconius, which departs from this norm in not being hyperparasitic. The presence of more diversified native parasitoids and hyperparasitoids from the Ichneumonoidea predicate a slow successive adaptation of the parasitoid fauna to the invasive host, Cameraria ohridella.     

Phytophthora ×alni and Phytophthora lacustris associated with common alder decline in Central Portugal

Since the early 1990s, an emerging disease induced by the highly aggressive oomycete Phytophthora ×alni has caused widespread alder decline across Europe. In parallel, P. lacustris, a recently described species associated with riparian habitats, has been subject of increasing interest. A field survey conducted in 2014 showed high mortality rates in alder stands located in the riparian gallery along two rivers in Central Portugal. The pathogens isolated from necrotic alder stem base during this study were identified as P. ×alni and P. lacustris. This paper is the first to report the occurrence of P. lacustris in Portugal and presents the first finding of P. ×alni affecting mature trees in natural ecosystems located in Central Portugal.     

Rescue of American chestnut with extraspecific genes following its destruction by a naturalized pathogen

Following the near-obliteration of American chestnut (Castanea dentata [Marsh.] Borkh.) by the chestnut blight early in the last century, interest in its restoration has been revived by efforts to develop a blight-resistant form of the species. We summarize progress and outline future steps in two approaches: (1) a system of hybridizing with a blight-resistant chestnut species and then backcrossing repeatedly to recover the American type and (2) transformation of American chestnut with a resistance-conferring transgene followed by propagation and conventional breeding. Several decades of effort have been invested in each approach. More work remains, but results indicate that success is within practical reach. The restoration of C. dentata to its native habitat now appears to be less a matter of time and conjecture than ever before in 90 years of work by public and private entities. The difficult and protracted task of incorporating extraspecific genes for resistance into a tree species with lethal susceptibility to a naturalized pathogen represents perhaps the most extreme of restoration challenges. Its pursuit by a small non-governmental organization supported primarily by philanthropy and volunteers may serve as a model for other species threatened by exotic pathogens or insects.     

Synergistic effect of drought and chestnut blight (Cryphonectria parasitica) on growth decline of European chestnut (Castanea sativa)

The chestnut blight fungus [Cryphonectria parasitica (Murill) Barr] has threatened European chestnut stands (Castanea sativa Mill.) in the 20th century, but infected trees recovered because of the appearance of hypovirulent strains. However, within a dry inner Alpine valley (Italy), blight‐infected C. sativa showing various degrees of crown dieback and dead trees were found. We conducted a dendroecological analysis to retrospectively evaluate a possible synergistic effect of blight infection in the early 1970s and climate stress on the growth decline of C. sativa. In the Eisack Valley (Italy), where annual precipitation is <700 mm, increment cores were taken from blight‐infected C. sativa (n = 103) showing different levels of decline symptoms, i.e. extent of crown dieback (healthy, moderate, severe), and from dead trees. Ring width and basal area increment (BAI) chronologies were developed based on dendroecological methods. Growth–climate relationships were explored using response function analysis and Pearson correlation coefficients. Major findings of our study were: (i) C. sativa growth is limited by low precipitation from December to February and high temperatures in June, (ii) BAI of all vitality classes except healthy trees shows a decreasing trend since 1980; and (iii) a severe drought in 1996 accelerated growth decline and caused death of infected C. sativa individuals. Because of the strong influence of climate on radial tree growth within the study area and observed divergent growth trends in selected vitality classes after infestation by hypovirulent C. parasitica strains in the early 1970s, we conclude that although blight infection is a prerequisite for observed growth declines, soil water availability in the years of drought strongly affects susceptibility to tree death.     

Pseudomonas syringae pv. aesculi associated with horse chestnut bleeding canker in Germany

Bacteria were isolated from necrotic lesions on a horse chestnut tree (Aesculus hippocastanum) with bleeding canker in Hamburg, Germany. Sequencing of the rDNA-ITS region revealed great similarity to pathovars of Pseudomonas syringae. Pseudomonas syringae pv. aesculi was identified by sequence homology of the gyrase B gene. This is the first report of P. syringae pv. aesculi in Germany. Phytophthora was not detected.     

Phytophthora dieback on narrow leaved ash in the Black Sea region of Turkey

Severe dieback symptoms were observed in a 490‐ha moist ash (Fraxinus angustifolia) lowland forest stand, comprising trees over 100 years old and in 100 ha of newly planted F. angustifolia near Sinop, in Turkey. Five of the 10 soil samples collected around stem bases of the diseased trees were baited using ash leaves and yielded a Phytophthora sp. This heterothallic species produced non‐caducous, non‐papillate sporangia in non‐sterile soil extract, and fluffy, even growth on corn meal and potato dextrose agars, and suppressed, even growth on grated carrot agar. Isolates were identified as Phytophthora taxon salixsoil based on internal transcribed spacer DNA sequences. This species has been recently redesignated as P. lacustris. Three isolates were found to be pathogenic when inoculated on the stem bases of three‐year‐old F. angustifolia saplings.     

Fast molecular detection of Pseudomonas syringae pv. aesculi in diseased horse chestnut trees

A molecular technique was used to detect the bacterium Pseudomonas syringae pv. aesculi in horse chestnut trees (Aesculus hippocastanum), affected by the recently recognized European ‘Pseudomonas horse chestnut bark disease’. The technique helped identify the pathogen within 6 h of sample preparation including DNA extraction, polymerase chain reaction (PCR) and electrophoresis until gel documentation. PCR primer pairs derived from the gyrase B gene sequence were used. Because of the great similarity in the gyrase B gene sequences of the numerous closely related P. syringae pathovars, the primers were not only totally specific to the pathovar aesculi, but also detected a few other pathovars. The assumption that other bacteria should not occur at least near to a necrotic lesion of a horse chestnut tree was corroborated by sequence identity of the PCR products obtained with the gyrase B gene sequence of P. syringae pv. aesculi. Koch’s postulates were fulfilled for an isolate of P. syringae pv. aesculi obtained from a diseased horse chestnut tree sampled in Hamburg in 2007.     

The oak decline: a new problem in Italy

The Authors observed, in Italy, over a number of years, isolated and grouped trees of the genus Quercus that declined over one or more vegetative seasons. The spread was very fast; in fact the trees declined in the course of 2–3 vegetative seasons. A series of symptoms was observed: production of epicormic shoots, yellowing of leaves, laceration of the bark, bleeding from the trunk. Many fungi were isolated: Armillaria spp., Cephalosporium spp., Cladosporium spp., Cylindrocarpon spp., Diplodia mutila, Hypoxylon mediterraneum, Phoma cavae, Pbomopsis quercina, Sporotrix spp. Future research will attempt to ascertain the possible causes of the decline.     

Vegetative compatibility types of Cryphonectria parasitica,causal agent of chestnut blight,in the Black Sea region of Turkey

Vegetative compatibility types (vc types) of 296 isolates of the chestnut blight fungus, Cryphonectria parasitica, were determined. The isolates had been obtained from 32 localities in 11 provinces in the Black Sea region of Turkey. Five vc types were detected: EU‐1, EU‐12, EU‐14, EU‐2 and EU‐5. The number of vc types found in single provinces varied between one and five. All of the five vc types were present only in the Kastamonu province. Vc type EU‐1 was detected in all the provinces. EU‐1 accounted for 90.8% of all isolates. Vc type EU‐12 was present in eight provinces and accounted for 6.8% of the isolates, whereas one or two isolates each of EU‐14, EU‐2 and EU‐5 were found in one or two provinces. Isolates possessing the white colony phenotype were considered to be hypovirulent. Hypovirulent isolates of each vc type were found, and they were detected in nine of 11 provinces.     

Recent spruce decline with biotic pathogen infestation as a result of interacting climate,deposition and soil variables

Decline or health deterioration of Norway spruce (Picea abies (L.) Karst.) dominated forest stands has recently been observed mainly in sub-mountainous parts of Central Europe. Forest inventory of 208 randomly distributed circular plots including field observations of spruce tree health and rot symptoms by honey fungus (Armillaria ostoyae) was used for assessing intensity of spruce forest health decline in a managed forest area of 12.7 th. ha located in Beskids Mts., NE Czech Republic. First, principal component analysis was used to separate inventory variables related to environmental stress (reduced apical increment, dry tree top and stem resin exudation due to A. ostoyae infestation) into PC1, and health deterioration symptoms associated with mechanical damage (peeling, crown breaks) into PC2. The first two principal components explained 59% of the total variability in health decline symptoms. Spatial variability of both principal components was explained using spatial lag regression model identified from a set of environmental variables including sulfur and nitrogen deposition, elevation, solar radiation, age of the forest stands and geological properties (geochemical reactivity index). Environmental stress (PC1) was associated with low elevations (sub-optimal for spruce), high level of nitrogen and sulfur deposition (their interaction), low geochemical reactivity and also stand age. On the other hand, mechanical damage (PC2) significantly increased with elevation and stand age. As the forest decline in Beskids Mts. is related to A. ostoyae spreading from local infestation hot spots, both principal components had a significant spatial autocorrelation, partly distorting the signal of environmental conditions. The results indicate that the disturbed forest soils by long-term acid deposition and subsequent nutrient degradation and more pronounced drought stress at low elevations are the most important drivers of the recent spruce health decline in Beskids Mts.     

Pestalotiopsis biciliata,a new leaf pathogen of Eucalyptus spp. recorded in Italy

The present paper describes new symptoms on Eucalyptus spp. in Central Italy associated with the fungal pathogen Pestalotiopsis biciliata. This fungal species has been previously recorded in Italy on grape associated with fruit rot. Artificial inoculation trials on leaves confirmed the pathogenicity of the fungus to Eucalyptus camaldulensis. P. biciliata is included in the list of Pestalotiopsis spp. responsible for leaf disease on eucalypts worldwide.     

Development of a quantitative real‐time PCR assay for the detection of Phytophthora austrocedrae,an emerging pathogen in Britain

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

Calonectria metrosideri,a highly aggressive pathogen causing leaf blight,root rot,and wilt of Metrosideros spp. in Brazil

The genus Metrosideros includes several tree, shrub and vine species, native to the Pacific Islands. Seedlings from 25 seed lots of Metrosideros polymorpha and two seed lots of M. tremuloides with symptoms of root rot, stem girdling, wilting and round, purple leaf spots were observed in the Forestry Nursery at the Universidade Federal de Viçosa, Brazil. In the original disease site, seedling mortality reached up to 71% in M. polymorpha and 34% in M. tremuloides. Single conidial cultures obtained from infected leaf, root and stem samples of M. polymorpha were used to identify the fungal species. Morphological characters and DNA sequences of four loci, containing partial sequences of β‐tubulin (TUB2), histone H3 (HIS3), calmodulin (CAL) and the elongation factor (tef‐1α) genes of three isolates, indicated that they belong to a new species, described here as Calonectria metrosideri sp. nov. Potting medium infestation and inoculation of seedlings of M. polymorpha with an inoculum suspension at 1 × 10⁴ conidia ml^?1 induced typical symptoms of the disease (leaf spots, root rot and wilt), similar to those observed under natural conditions. Calonectria metrosideri was re‐isolated, which fulfilled Koch's postulates, and confirmed its status as a pathogen.     

Involvement of Phytophthora species in white oak (Quercus alba) decline in southern Ohio

This study was initiated to investigate the possible role of Phytophthora species in white oak decline (Quercus alba) in southern Ohio at Scioto Trail State Forest. Surveys demonstrated the presence of four species of Phytophthora including one novel species. By far, the most common species was P. cinnamomi; P. citricola and P. cambivora were isolated infrequently. In few instances, P. cinnamomi was isolated from fine roots and necroses on larger roots. No special pattern of incidence was found, but P. cinnamomi was more commonly isolated from greater Integrated Moisture Index values suggesting moist lower bottomlands favour this Phytophthora species. When tree crown condition was examined relative to the presence of Phytophthora, no significant association was found. However, roots of declining P. cinnamomi‐infested trees had 2.5 times less fine roots than non‐infested and healthy trees, which was significantly different. The population densities of P. cinnamomi from declining trees were significantly greater than from healthy trees, suggesting increased pathogen activity that has the potential to cause dieback and decline and possibly the cause of a reduced fine root amount found on declining trees.     

Incidence of Phytophthora species in oak forests in Austria and their possible involvement in oak decline

A survey on the occurrence of Phytophthora species in oak ecosystems in Austria was conducted from April to May 1999 and in June 2000. The investigations were carried out at 35 study sites distributed throughout the zone of oak forests in eastern Austria. Four oak species, including Quercus robur, Q. petraea, Q. cerris and Q. pubescens were considered in the survey. Rhizosphere soil samples were taken from sample trees, which consisted of healthy and declining trees as indicated by their crown transparency. Young oak leaflets were used as baits to recover Phytophthora species. The assemblage of Phytophthora spp. detected in Austrian oak forests consisted of five species, including Phytophthora quercina, P. citricola, P. gonapodyides, P. europaea and P. syringae. P. quercina and P. citricola were isolated from 11 and seven sites, respectively, and were thus the most common and most widely distributed species. The three other species were recovered only sporadically. P. citricola could be separated into two morphologically and genetically well‐characterized types (A and B). Phytophthora species, in particular the common P. quercina and P. citricola occurred on sites showing a wide variety of soil types, soil textures and moisture classes. There was mild evidence for connection between deteriorating crown status and the presence of Phytophthora spp. Furthermore, significant differences in contents of magnesium, as well as calcium, aluminium, nitrogen and carbon at different soil depths (0–10, 10–20 and 20–40 cm) were detected between Phytophthora‐infested and Phytophthora‐free sites. The results of the present study provide circumstantial evidence that Phytophthora species are involved in oak decline at certain sites in Austria.     

Beech decline in Central Europe driven by the interaction between Phytophthora infections and climatic extremes

During the past decade, and in particular after the wet year 2002 and the dry year 2003, an increasing number of trees and stands of European beech (Fagus sylvatica L.) in Bavaria were showing symptoms typical for Phytophthora diseases: increased transparency and crown dieback, small‐sized and often yellowish foliage, root and collar rot and aerial bleeding cankers up to stem heights of >20 m. Between 2003 and 2007 134 mature beech stands on a broad range of geological substrates were surveyed, and collar rot and aerial bleeding cankers were found in 116 (86.6%) stands. In most stands the majority of beech trees were declining and scattered or clustered mortality occurred. Bark and soil samples were taken from 314 trees in 112 stands, and 11 Phytophthora species were recovered from 253 trees (80.6%) in 104 stands (92.9%). The most frequent species were P. citricola, P. cambivora and P. cactorum. Primary Phytophthora lesions were soon infected by a series of secondary bark pathogens, including Nectria coccinea, and wood decay fungi. In addition, infected trees were often attacked by several bark and wood boring insects leading to rapid mortality. Bark necroses were examined for their probable age in order to determine whether the onset of the current Phytophthora epidemic was correlated to rainfall rates recorded at 22 Bavarian forest ecosystem monitoring stations. A small‐scale survey in nine Bavarian nurseries demonstrated regular infestations of all beech fields with the same range of Phytophthora species. The results indicate that (1) Phytophthora species are regularly associated with beech decline and may also be involved in the complex of ‘Beech Bark Disease’, (2) excessive rainfalls and droughts are triggering the disease, and (3) widespread Phytophthora infestations of nursery stock might endanger current and future silvicultural projects aiming on the replacement of non‐natural conifer stands by beech dominated mixed stands.