Trunk Pathogens Research Articles

Fungal trunk pathogens associated with wood decay of almond trees on Mallorca (Spain).

Severe decline of almond trees has recently been observed in several orchards on the island of Mallorca (Balearic Islands, western Mediterranean Sea). However, the identity of the causal agents has not yet been investigated. Between August 2008 and June 2010, wood samples from branches of almond trees showing internal necroses and brown to black vascular streaking were collected in the Llevant region on the island of Mallorca. Several fungal species were subsequently isolated from the margin between healthy and symptomatic tissue. Five species of Botryosphaeriaceae (namely Botryosphaeria dothidea, Diplodia olivarum, D. seriata, Neofusicoccum australe and N. parvum), Eutypa lata, Phaeoacremonium iranianum and Phomopsis amygdali were identified based on morphology, culture characteristics and DNA sequence comparisons. Neofusicoccum parvum was the dominant species, followed by E. lata, D. olivarum and N. australe. First reports from almond include D. olivarum and Pm. iranianum. Two species are newly described, namely Collophora hispanica sp. nov. and Phaeoacremonium amygdalinum sp. nov.

Fungi associated with die-back symptoms of apple and pear trees, a possible inoculum source of grapevine trunk disease pathogens

A survey was undertaken on apple and pear trees in the main pome fruit growing areas ofthe Western Cape of South Africa to determine the aetiology of trunk diseases with specific reference to pathogens known to occur on grapevine, which are frequently cultivated in close proximity of these orchards. Several fungal genera containing known trunk disease pathogens were found. Two Diplodia species, D. seriata and Diplodia sp., were isolated along with Neofusicoccum australe and N. vitifusiforme . Four Phaeoacremonium species, Phaeoacremonium aleophilum , Pm. iranianum , Pm. mortoniae and Pm. viticola , two Phomopsis species linked to clades identified in former studies as Phomopsis theicola and Phomopsis sp. 7, and Eutypa lata were found. In addition, Paraconiothyrium brasiliense , Pr. variabile and a Pyrenochaeta like species were also isolated. Diplodia seriata (56% of total isolates) and Pm. aleophilum (22%) were most frequently isolate. First reports from pear wood include the Phaeoacremonium spp. and Paraconiothyrium brasiliense , while new reports from apple include Pm. aleophilum , Ph. theicola , Phomopsis sp. 7, Pr. variabile and E. lata . A pathogenicity trial was undertaken to determine the role of these species on apple, pear and grapevine shoots. Neofusicoccum australe caused the longest lesions on grapevine shoots, while Pr. variabile , D. seriata , Pm. mortoniae and the Pyrenochaeta -like sp. caused lesions that were longer than non-inoculatedand non-pathogen experimental controls. On pear shoots, Diplodia sp. and N. australe caused the longest lesions,followed by D. seriata and E. lata . On apple shoots, the longest lesions were caused by N. australe and Pm. iranianum . These results demonstrate that apple and pear trees in Western Cape orchards are hosts to many known trunk pathogens along with potential new trunk disease-causing fungi.

Temporal susceptibility of grapevine pruning wounds to trunk pathogen infection in South African grapevines.

Grapevine trunk diseases, caused by a range of phytopathogenic fungi, represent a serious impediment to wine and table grape production wherever these crops are cultivated. Previous studies have shown that the distribution of these pathogens is influenced by climate and that they are associated with a variety of internal wood decay symptoms. Little was known, however, about the influence of climate on the disease symptomatology of the different pathogens in a specific area. To address this, a survey was conducted in 30 wine and table grape vineyards in summer, marginal and winter rainfall regions of South Africa. Apart from Eutypa lata , which occurred only in the winter rainfall region, Phaeomoniella chlamydospora , species of Phaeoacremonium , Botryosphaeriaceae and Phomopsis occurred in all regions surveyed. The incidence of the fungal genera and species associated with trunk disease varied between regions, with overlapping symptom profiles that differed based on the climatic region. These findings suggest that symptom-based disease diagnosis alone is unreliable and that distribution and symptomatology of grapevine trunk pathogens are strongly influenced by climatic conditions in a specific production region.

Grapevine cultivar variation to pruning wound protection by Trichoderma species against trunk pathogens

Using Trichoderma species to protect grapevine pruning wounds from trunk disease pathogensis one of the options available for managing grapevine trunk diseases. The growth and persistence of Trichoderma species in the pruning wound and the resulting control effect may depend on intrinsic wound factors and hence may vary between cultivars. Cultivar variability to pruning wound protection by Trichoderma species was evaluated in eight wine grape (Cabernet Sauvignon, Chardonnay, Chenin blanc, Colombar, Merlot,Pinotage, Sauvignon blanc and Shiraz) and four table grape (Prime, Red Globe, Thompson Seedless andVictoria) cultivars. Two strains of Trichoderma atroviride (USPP-T1 and USPP-T2) separately or in combinationand Eco 77® a registered pruning wound biocontrol agent based on T. harzianum , were applied to fresh pruning wounds of spur-pruned wine grapevines and cane-pruned table grapevines. Trichoderma spp. and a variety of trunk pathogens, Phaeomoniella chlamydospora and species of Phaeoacremonium , Phomopsis , Botryosphaeriaceae and Diatrypaceae, were isolated from the pruning wounds eight months after treatment. Significant treatment × cultivar interactions (P 0.50;P<0.05) though not in Chardonnay (r=0.37; P=0.11), Pinotage (r=-0.12; P=0.62), Sauvignon blanc (r=0.26;P=0.26) and Victoria (r=0.29; P=0.22). It was concluded that the wound protection effect of Trichoderma spp.is also dependent on the Trichoderma -grapevine interaction and is not only due to the suppressive effect of Trichoderma spp. on the pathogens.

Evaluation of biocontrol agents for grapevine pruning wound protection against trunk pathogen infection.

Trunk diseases of grapevine are caused by numerous pathogens, including Eutypa lata , Phaeomoniella chlamydospora , and species of Botryosphaeriaceae (incl. Botryosphaeria and aggregate genera), Phomopsis and Phaeoacremonium . Since infections occur mainly through pruning wounds, that have been shown by previous research to stay susceptible for up to 16 weeks after pruning, long-term pruning wound protection is required for prevention of infection. This study evaluated several biocontrol agents against a range of trunk disease pathogens in dual plate laboratory trials to determine macroscopic and microscopic interactions. The biocontrol agents had a substantial effect on all the pathogens, with a wide range of macroscopic and microscopic interactions observed. The best performing biocontrol agents were tested in two field trials. Fresh pruning wounds were treated with benomyl, Trichoderma products (Biotricho®, Vinevax® and ECO 77®) and isolates (USPP-T1 and -T2, identified as T. atroviride ) and Bacillus subtilis . Seven days after treatment the pruning wounds were inoculated by spraying with spore suspensions of Neofusicoccum australe , N. parvum , Diplodia seriata , Lasiodiplodia theobromae , Eutypa lata , Phaeomoniella chlamydospora or Phomopsis viticola . Eight months after inoculation, the treatments were evaluated by isolation onto potato dextrose agar. The efficacy of the biocontrol agents was in most cases similar or superior to that observed for benomyl. Isolate USPP-T1, in particular, was very effective, reducing incidence of Ph. viticola , E. lata , Pa. chlamydospora , N. australe , N. parvum , D. seriata and L. theobromae by 69, 76, 77, 78, 80, 85 and 92%, respectively. This is the first report of biological protection of grapevine pruning wounds against this group of grapevine trunk disease pathogens.

The distribution and symptomatology of grapevine trunk disease pathogens are influenced by climate

Grapevine trunk diseases, caused by a range of phytopathogenic fungi, represent a serious impediment to wine and table grape production wherever these crops are cultivated. Previous studies have shown that the distribution of these pathogens is influenced by climate and that they are associated with a variety of internal wood decay symptoms. Little was known, however, about the influence of climate on the disease symptomatology of the different pathogens in a specific area. To address this, a survey was conductedin 30 wine and table grape vineyards in summer, marginal and winter rainfall regions of South Africa. Apart from Eutypa lata , which occurred only in the winter rainfall region, Phaeomoniella chlamydospora , species of Phaeoacremonium , Botryosphaeriaceae and Phomopsis occurred in all regions surveyed. The incidence of the fungal genera and species associated with trunk disease varied between regions, with overlapping symptom profiles that differed based on the climatic region. These findings suggest that symptom-based disease diagnosis alone is unreliable and that distribution and symptomatology of grapevine trunk pathogens are strongly influenced by climatic conditions in a specific production region.

Susceptibility of grapevine tissues to Neofusicoccum luteum conidial infection

This study investigated the ability of Neofusicoccum luteum to infect wounded shoots, trunks, pruned cane ends, leaf surfaces, buds, berries and roots, and its further progression into stem tissues. All tissue types were susceptible to infection except roots, with highest incidences in trunks (100%), cane ends (100%), shoots (92%) and buds (88%), indicating that in New Zealand, N. luteum is primarily a trunk and shoot pathogen. In trunks, there were no external symptoms, although N. luteum could be reisolated from 60 to 70 cm acropetally from the inoculation site after 4 months, by which time the pathogen had progressed into side shoots which became necrotic. Wounded and non‐wounded buds became infected; most were killed, with basipetal progression of the pathogen into the supporting shoots. Berries wounded and inoculated at the pre‐bunch closure stage were susceptible to N. luteum infection, with isolation incidence increasing over the season and peaking at harvest, when infected berries became mummified and produced pycnidia with many conidia. The pathogen was also able to progress from berries into bunch stems and supporting canes. Results from this research have indicated that N. luteum infection can occur in all aerial grapevine tissues and progress to young stem tissues where it causes wood necrosis. Growers should remove mummified berries from vineyard trash to ensure that pruning and trimming times do not coincide with rainy periods when conidia are released and dispersed. Furthermore, the susceptibility of buds to N. luteum infection indicates the need for fungicide sprays before budburst in spring.

Fungal Trunk Pathogens in the Grapevine Propagation Process: Potential Inoculum Sources, Detection, Identification, and Management Strategies

Fungal Trunk Pathogens in the Grapevine Propagation Process: Potential Inoculum Sources, Detection, Identification, and Management Strategies

Detection of botryosphaeriaceous species in environmental samples using a multi‐species primer pair

Botryosphaeriaceous species are significant grapevine trunk pathogens worldwide, which can be difficult to identify to species level using conventional morphological methods. This study developed and optimized a quick, reliable molecular identification method that could facilitate investigations into the epidemiology of these diseases in vineyards. The multi‐species primers, BOT100F and BOT472R, amplified a 371–372 bp portion of the rRNA gene region from the six botryosphaeriaceous species commonly found in New Zealand vineyards. In silico analysis indicated that they would amplify DNA from six of the 12 lineages of the Botryosphaeriaceae, including all of the main species pathogenic to grapevines. A detection sensitivity of 1 and 0·1 pg DNA in standard and nested PCR, respectively, was achieved and this was calculated as equivalent to 2·5 conidia. Validation of the primers for environmental samples showed that their specificity was not compromized by the presence of competing DNA templates extracted from wood and soil. Single stranded conformational polymorphism (SSCP) analysis of the amplicons could resolve Neofusicoccum australe, N. luteum, Diplodia mutila and D. seriata, but did not differentiate between N. parvum and N. ribis. The optimized PCR‐SSCP was used to identify botryosphaeriaceous species present in rainwater traps collected over 1 year in a vineyard known to contain infected vines. It could detect multiple species in individual samples and demonstrated differences in the dispersal patterns of conidia from different species. Given the specificity and sensitivity of this method it could prove useful in epidemiology studies involving the numerous botryosphaeriaceous species that infect a wide range of host species.

Effects of Soil-Applied and Trunk and Petiole-Injected Manganese on Manganese Content of Coconut Palm (Cocos nucifera)

Manganese deficiency is a common and potentially fatal disorder of palms growing in highly leached and calcareous soils. Soil applications of MnSO4 may not always be effective in treating this disorder due to rapid oxidation of Mn to less available forms. Trunk injection with MnSO4 (2.0 g Mn) was found to be more effective in increasing foliar Mn concentrations than soil application (192 g Mn) or petiole injections with 0.1 g Mn in a single hole, or 0.5 g Mn divided among four holes. In contrast to trunk injection, neither petiole injections nor soil application of MnSO4 increased foliar Mn concentrations above that of the untreated control palms. Trunk injections, while effective, result in permanent wounds that could potentially serve as entry sites for the trunk pathogen Thielaviopsis paradoxa, especially on young palms with minimal trunks.

First Report of Lasiodiplodia crassispora as a Pathogen of Grapevine Trunks in South Africa.

In 2003 and 2004, a survey of grapevine (Vitis vinifera L.) trunk pathogens was conducted in 30 vineyards in the Western and Northern Cape and Limpopo provinces of South Africa. In each vineyard, 20 visually healthy plants were sampled randomly by removing the distal part of one cordon arm. Isolations were made onto potato dextrose agar (PDA) from the internal wood decay symptoms observed in the cordon samples. Seven Botryosphaeriaceae spp. were identified, including Lasiodiplodia crassispora (1). Other Botryosphaeriaceae spp. are known grapevine trunk pathogens (2). Species identity was confirmed by DNA sequence data of the partial translation factor 1-α gene (1) and sequences deposited in GenBank (GU233658 and GU233659). The L. crassispora isolates (CBS 125626 and 125627) were associated with brown internal necrosis, a known symptom of grapevine Botryosphaeriaceae spp. infection (3), in the cordon arms of Ruby Cabernet grapevines occurring in two vineyards in the Northern Cape Province. L. crassispora was described from cankered wood of Santalum album in Western Australia and endophytically from Eucalyptus urophylla in Venezuela (1). Its grapevine pathogen status was determined using both isolates in a repeated pathogenicity test that included three isolates each of Botryosphaeria dothidea and Neofusicoccum australe as positive controls (2), Trichoderma harzianum as a nonpathogen treatment, and an uncolonized agar plug as a negative control. The Botryosphaeriaceae spp. and T. harzianum were plated on PDA and incubated at 25°C for 7 days. Lignified, 6-month-old shoots of grapevine cv. Chardonnay were excised from grapevines with internodes 4 to 6 used for inoculations. Before wounding, shoots were disinfected by submersion for 1 min in a 1 ml/liter solution of a quaternary ammonium compound (Sporekill; ICA International Chemicals (Pty) Ltd, Stellenbosch, South Africa). Twelve shoots were used for each isolate or control treatment. Wounds were made 2 mm deep on the fifth internode of the shoots with a 5-mm flame-sterilized cork borer (2,3). Wounds were inoculated with a pathogen colonized agar plug (5 mm in diameter) or an uncolonized agar plug and then covered with Parafilm (2,3). Inoculated shoots were incubated in the dark in moist chambers for 14 days at 25°C. After incubation, the bark of the shoots was peeled from the area around the wound and the lengths of any resultant lesions were measured under sterile conditions. The inoculum effect was assessed by analysis of variance and Student's t-test. Results showed that significantly (P < 0.0001) longer lesions were caused by L. crassispora (13.36 mm) compared with N. australe (9.27 mm) and B. dothidea (5.28 mm) and also significantly longer than lesions caused by the nonpathogen and negative controls (3.23 and 2.90 mm, respectively). To determine if lesions were caused by inoculated fungi, isolations were made from the tissue at the edges of the lesions by aseptically removing five 0.5 × 1 mm pieces of wood and placing them on PDA dishes amended with 0.04 g/liter of streptomycin sulfate. Dishes were incubated under normal fluorescent light at 25°C for 14 days before identifying isolated fungi based on morphological and cultural characteristics (1). To our knowledge, this is the first report of L. crassispora as a grapevine pathogen.

Rethinking the value of high wood density

Summary 1. Current thinking holds that wood density mediates a tradeoff between strength and economy of construction, with higher wood density providing higher strength but at higher cost. 2. Yet the further away wood fibres are from the central axis of the trunk, the more they increase the strength of the trunk; thus, a fat trunk of low‐density wood can achieve greater strength at lower construction cost than a thin trunk of high‐density wood. 3. What then are the countervailing advantages of high wood density? 4. We hypothesize that high wood density is associated with lower maintenance costs due to lower trunk surface area, as surface area correlates with maintenance respiration. 5. This advantage would be particularly important to long‐lived trees and could in part explain why they tend to have high wood density. 6. High wood density has also been associated with lower risk of trunk breakage, xylem implosion and pathogen invasion, but we argue that these relationships are not causal and instead reflect correlated selection on other traits of value to long‐lived trees. 7. This revaluation of the costs and benefits of high wood density has important implications for understanding tree life‐history evolution, functional diversity, forest carbon stocks and the impacts of global change.

Temporal spore dispersal patterns of grapevine trunk pathogens in South Africa

Trunk disease pathogens of grapevines, viz. Phaeomoniella chlamydospora, Eutypa lata and several species in Botryosphaeriaceae, Phaeoacremonium and Phomopsis are known to infect fresh pruning wounds by means of air-borne inoculum released after rainfall or prolonged periods of high relative humidity. Recent surveys have demonstrated that most or all of these pathogens are present in climatically diverse grape growing regions of South Africa. However, the factors controlling spore dispersal of these pathogens in vineyards were largely unknown. To address this question, spore trapping was done in a Chenin Blanc vineyard in the Stellenbosch area, South Africa, for 14 weeks during the grapevine pruning period from June to mid-September of 2004 and 2005. Hourly recordings of weather data were done by a weather station in the row adjacent to the spore trap. Spores of E. lata and Phomopsis and species in Botryosphaeriaceae were trapped throughout the trapping periods of 2004 and 2005, with higher levels of trapped spores recorded in 2005. The spores of all three pathogens were trapped during or after periods of rainfall and/or high relative humidity. In neither of the 2 years were spores of Pa. chlamydospora or Phaeoacremonium spp. trapped. Results indicated that spore event incidence, as well as the amount of spores released during a spore event of above-mentioned pathogens, were governed by rainfall, relative humidity, temperature and wind speed prior to and during the spore events.

Effect of hot‐water treatments in vitro on conidial germination and mycelial growth of grapevine trunk pathogens

In this study, the sensitivity of Cadophora luteo‐olivacea, Cylindrocarpon liriodendri, Cn. macrodidymum and eight species of the genus Phaeoacremonium to hot‐water treatments (HWTs) in vitro was evaluated. Conidial suspensions and plugs of agar with mycelia were placed in Eppendorf vials and incubated for 30, 45 or 60 min in a hot‐water bath at 41, 42, 43, 44, 45, 46, 47, 48 or 49°C for Cylindrocarpon spp. and at 49, 50, 51, 52, 53, 54 or 55°C for Ca. luteo‐olivacea and Phaeoacremonium spp. In general, conidial germination and the colony growth rate of all pathogens decreased with increased temperature and time combinations. Cylindrocarpon spp. were more sensitive than Ca. luteo‐olivacea and Phaeoacremonium spp. to HWT temperatures. Conidial germination of Ca. luteo‐olivacea was inhibited by treatments above 51°C–30 min, while treatments up to 54°C–60 min were necessary to inhibit the mycelial growth. For Cylindrocarpon spp., conidial germination was inhibited by treatments above 45°C–45 min, while treatments above 48°C–45 min were necessary to inhibit the mycelial growth. Regarding Phaeoacremonium spp., treatments up to 54°C–60 min were necessary to completely inhibit both conidial germination and mycelial growth. These results suggest that current HWT protocols at 50°C for 30 min may be sufficient to control Cylindrocarpon spp. However, it would be necessary to develop HWT using higher temperatures to reduce the incidence of Ca. luteo‐olivacea and Phaeoacremonium spp. infections.

Evaluation of the grapevine nursery propagation process as a source of Phaeoacremonium spp. and Phaeomoniella chlamydospora and occurrence of trunk disease pathogens in rootstock mother vines in Spain

Five commercial nurseries were sampled in 2007 to evaluate the grapevine nursery propagation process as a source of Petri disease pathogens (Phaeoacremonium spp. and Phaeomoniella chlamydospora). Samples were taken at four stages of the propagation process: pre-grafting hydration tanks, scissors used for cutting buds, grafting machines and peat used to promote root development. All samples were analysed using two different techniques: nested PCR using specific primers for Phaeoacremonium spp. (Pm1/Pm2) and Pa. chlamydospora (Pch1/Pch2); and fungal isolation by culturing on semi-selective medium. Either Phaeoacremonium spp. or Pa. chlamydospora were detected at any of these stages, and more importantly they were viable since they were detected by isolating on culturing medium. Additionally, the importance of grapevine rootstock mother fields as sources of inoculum in the nurseries was studied. Fourteen grapevine rootstock mother fields were surveyed in 2006 and 2007 for the occurrence of fungal trunk pathogens. A total of 16.4% and 30% of the plants sampled in 2006 and 2007, respectively were infected. Petri disease pathogens (Pa. chlamydospora, Phaeoacremonium aleophilum, Pm. parasiticum) and several Botryosphaeriaceae species (Neofusicoccum parvum, Botryosphaeria dothidea, Lasiodiplodia theobromae, N. australe, N. mediterraneum and N. vitifusiforme) and Phomopsis viticola were isolated. This is the first time N. mediterraneum has been isolated from grapevines and the first report of N. australe, N. mediterraneum and N. vitifusiforme in Spain. This work shows that grapevine rootstock mother plants and the propagation process of grapevine plants should be considered as important sources of inoculum for fungal trunk pathogens, and especially of Petri disease pathogens.

A Multiplex PCR System for the Specific Detection of Cylindrocarpon liriodendri, C. macrodidymum, and C. pauciseptatum from Grapevine

Cylindrocarpon liriodendri and C. macrodidymum are the causal agents of grapevine black foot disease. Recently, a third species, C. pauciseptatum, has been isolated from roots of grapevine showing decline symptoms. Currently, reliable identification of isolates of these species through phenotypical characteristics has not been possible. The polymerase chain reaction (PCR)-based method developed in this study allows a quick and easy detection of Cylindrocarpon spp. associated with grapevine. Three primer pairs annealing to variable, partly species-specific sites of the internal transcribed spacer regions amplified species-specific PCR fragments of different sizes in C. liriodendri, C. macrodidymum, and C. pauciseptatum in a multiplex assay with DNA obtained with both quick and traditional extraction methods. They did not generate any PCR product in other fungal trunk pathogens or contaminants commonly associated with grapevines. When universal fungal ITS primers were used in a nested multiplex PCR, the three primer pairs also detected C. liriodendri, C. macrodidymum, and C. pauciseptatum in total DNA extracted from roots of inoculated grapevines. The designed methods can be used for the diagnosis of these fungi from pure culture or infected grapevines.

Occurrence of Fungal Pathogens Associated with Grapevine Nurseries and the Decline of Young Vines in Spain

AbstractThe occurrence of fungal grapevine trunk pathogens associated with grapevine nurseries and the decline of young vines in Spain was determined in extensive surveys conducted in nurseries and vineyards with young plants. The presence of Phaeomoniella chlamydospora, Phaeoacremonium aleophilum, Cylindrocarpon spp., Botryosphaeria obtusa and Botryosphaeria spp. was detected in all the surveys that were carried out. This study provides evidence for the presence of these pathogens in Spanish grapevine nurseries. Cylindrocarpon spp., B. obtusa and Botryosphaeria spp., were isolated very early in the planting material production process, nevertheless, P. aleophilum and P. chlamydospora were not detected until the rootstock–scion combinations were planted in the field to develop shoots and roots during summer. The occurrence of trunk disease pathogens in symptomatic young vineyards was also high, suggesting that infected plant material might have been used. Phaeomoniella chlamydospora and P. aleophilum were the main species associated with the decline of young vines, followed by Botryosphaeria spp., Cylindrocarpon spp. and B. obtusa. This research confirmed the importance of fungal grapevine trunk pathogens associated with the decline of young vineyards in Spain and suggested the connection between the presence of these fungi in the nurseries and the disease in the fields.

Identification of Eutypa lata by PCR-RFLP.

Eutypa lata is a vascular canker pathogen of woody plants commonly diagnosed by isolating the pathogen from infected tissue. Related fungi from the same family, the Diatrypaceae, also have been found in association with grapevine in Californian vineyards. An in situ polymerase chain reaction (PCR) method has been developed for detection of E. lata in infected wood tissue. However, our results indicate that this method also would amplify other Diatrypaceous fungi, which could potentially lead to an incorrect diagnosis. Therefore, we developed a PCR-restriction fragment length polymorphism (PCR-RFLP) assay. The internal transcribed spacer (ITS)1/5.8S/ITS2 ribosomal DNA region was amplified by PCR using universal primers, and RFLP patterns were determined after digestion with AluI. The restriction profiles obtained served to distinguish E. lata from wood trunk pathogens of grapevine (Phomopsis viticola, Botryodiplodia sp., Phaeoacremonium aleophilum, and Phaeomoniella chlamydospora), Diatrypaceous fungi (Diatrype sp., Diatrypella sp., Eutypella vitis, and Eutypa leptoplaca), and Cryptovalsa sp. found on dead wood of grapevine, and other Eutypa spp. (E.petrakii var. hederae, E. astroidea, E. crustata, and E. lejoplaca), with the exception of E. armeniacae, which we regard as a synonym for E. lata, and E. laevata, which has not been found on grapevine.