A serious canker disease caused by Immersiporthe knoxdaviesiana gen. et sp. nov. (Cryphonectriaceae) on native Rapanea melanophloeos in South Africa

Recent disease surveys in the Western Cape province of South Africa have revealed a previously unknown and serious stem canker disease on native Rapanea melanophloeos (Myrsinaceae, Ericales) trees. Cankers commonly result in the death of branches or entire stems. Fruiting structures typical of fungi in the Cryphonectriaceae were observed on the surfaces of cankers. In this study, the fungus was identified and its pathogenicity to R. melanophloeos was tested. Multigene phylogenetic analyses based on DNA sequences of the partial LSU gene, ITS region of the nuclear ribosomal DNA gene and two regions of the β‐tubulin (BT) gene, showed that the fungus represents a formerly undescribed genus and species in the Cryphonectriaceae. The fungus was also morphologically distinct from other genera in this family. Inoculation trials showed that the fungus described here as Immersiporthe knoxdaviesiana gen. et sp. nov. is an aggressive pathogen of R. melanophloeos trees.     

Corticimorbus sinomyrti gen. et sp. nov. (Cryphonectriaceae) pathogenic to native Rhodomyrtus tomentosa (Myrtaceae) in South China

The Cryphonectriaceae includes many important tree pathogens, especially of the Myrtales. Disease surveys on Myrtales in South China revealed a stem canker disease on native Rhodomyrtus tomentosa (Myrtaceae, Myrtales) trees in the proximity of Eucalyptus plantations in GuangXi Province and in a natural forestry area in the Hong Kong Region. Fruiting structures with typical characteristics of the Cryphonectriaceae were observed on the canker surfaces. The fungus was identified based on DNA sequence comparisons and morphological features, and its pathogenicity was tested on R. tomentosa under field conditions. DNA sequence comparisons for the partial large subunit, partial internal transcribed spacer nuclear ribosomal DNA, and two regions of the β‐tubulin gene showed that the fungus represents a previously undescribed genus and species in the Cryphonectriaceae. Based on morphology, the fungus is most similar to species in the genus Chrysoporthe, but can be distinguished from this and the other genera of Cryphonectriaceae. The fungus provided with the name Corticimorbus sinomyrti gen. et sp. nov., is described. Field inoculations showed that it is highly pathogenic to R. tomentosa trees, with the ability to kill inoculated branches within 4 weeks. Pathogenicity tests also showed that C. sinomyrti is pathogenic to a tested Eucalyptus clone. Pathogenicity on both native R. tomentosa and non‐native Eucalyptus suggests that this fungus should be monitored carefully to limit its possible spread to commercially grown Eucalyptus in South China.     

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.     

Multilocus PCR assay reveals high diversity of vegetative compatibility types in populations of Cryphonectria parasitica in Croatia

The ascomycete fungus Cryphonectria parasitica, causal agent of chestnut blight, is probably one of the best known invasive fungal pathogens in forests of Europe and North America. Mycovirus that reduces virulence of C. parasitica can be used as a biocontrol agent of the chestnut blight. However, anastomosis‐mediated virus transmission is limited by a vegetative (in)compatibility (vc) system involving at least six known diallelic vic genetic loci. This study looked at vegetative compatibility (vc) diversity in two populations of C. parasitica in Croatia. For that purpose, a PCR assay was validated and implemented using already known/published and newly designed primers for amplification of six known vic loci. The vc genotypes determined by PCR for 158 C. parasitica isolates investigated in this study were in complete agreement with the vc genotypes determined by pairwise co‐culturing of the same isolates, revealing the specificity and accuracy of the PCR‐based molecular vic genotyping assay. Twenty‐six unique vc genotypes were found among 158 isolates, and 19 vc types per population, which makes Croatian C. parasitica populations among the most diverse in Europe regarding the number of vc types and genetic diversity. Low values of multilocus linkage disequilibrium suggest sexual reproduction as a major contributor to high C. parasitica genetic diversity in studied populations.     

First record of Dryocosmus kuriphilus in Greece

After the first introduction of chestnut gall wasp, Dryocosmus kuriphilus, in Italy, it has spread rapidly in several other European countries, probably largely due to trade in chestnut plants. Despite the measures taken to prevent further spread of this pest to other countries, D. kuriphilus was detected in Greece for the first time in 2014. However, surveys have shown that the pest is still of very limited distribution in Greece. Having faced the damage caused by chestnut blight (Cryphonectria parasitica) over the last 50 years, the production of chestnuts in Greece is now seriously threatened by chestnut gall wasp, which has already caused extensive losses in chestnut production of other countries. In order to minimize the impact of chestnut gall wasp, control measures are being designed involving the release of the parasitoid Torymus sinensis.     

Chestnut blight in Yugoslavia1

The present state of chestnut blight caused by the fungus Cryphonectria parasitica in Yugoslavia is presented. Canker disease is present all over the country wherever chestnut stands occur. Field observations have shown the presence of abnormal disease symptoms at some localities. Isolates of C. parasitica from these sites varied widely in cultural characteristics and pathogenic properties, some showing the characteristics of hypovirulent strains.     

Chestnut blight fungus in Croatia: diversity of vegetative compatibility types,mating types and genetic variability of associated Cryphonectria hypovirus 1

In order to improve understanding of its diversity, 338 isolates of Cryphonectria parasitica, the causal agent of chestnut blight, were sampled from 10 chestnut populations throughout chestnut‐growing coastal and continental areas of Croatia. Eighteen vegetative compatibility (VC) types were identified. The VC type EU‐1 was the most widespread, comprising 42·9% of the isolates, followed by EU‐2 (21%) and EU‐12 (14·2%). In respect to the occurrence of the main VC types, the C. parasitica populations in Croatia combined features of both northwestern and southeastern European populations. Perithecia and mating‐type ratios of approximately 1 : 1 were found in all populations, suggesting that sexual reproduction of the fungus is common in Croatia. Natural hypovirulence was also evident in all populations, with incidence of hypovirus‐infected isolates ranging from 12·7% in Istria‐Buje to 66·6% in the continental part of the country. A total of 36 hypovirus‐infected isolates sampled throughout Croatia were analysed in ORF‐A and ORF‐B by RT‐PCR/RFLP analysis. All viral isolates belonged to the Italian subtype of Cryphonectria hypovirus 1 (CHV‐1) and were closely related to the isolates found in other European countries. The RFLP patterns found were also identical or similar to the patterns of three isolates collected in Croatia 22 years ago, suggesting a slow evolution of the hypovirus.     

A new genus of Cryphonectriaceae isolated from Lagerstroemia speciosa in southern China

Numerous species in the Cryphonectriaceae have been recorded on the Myrtales and many of these are economically important pathogens of Eucalyptus. Some species have also recently been shown to be endophytes of native Myrtaceae and to have undergone host jumps to infect Eucalyptus species established as exotics in plantations. Recent surveys in the GuangDong and HaiNan Provinces of South China reveal the presence of a species of Cryphonectriaceae associated with cankers on trees of Lagerstroemia speciosa (Lythraceae, Myrtales). Fungal structures were observed on the surface of dead bark covering cankers and on branch stubs. Multigene phylogenetic analyses were conducted based on DNA sequence comparisons of the partial LSU gene, ITS region of the nuclear ribosomal RNA gene and two regions of the β‐tubulin (BT) gene. The results revealed the presence of a previously undescribed genus and species in the Cryphonectriaceae. The fungus is described here as Chrysomorbus lagerstroemiae gen. et sp. nov. Inoculation tests showed that it is an aggressive pathogen on L. speciosa and that it can also infect Eucalyptus.     

First report of the canker pathogen Endothia gyrosa on Eucalyptus in South Africa

During country-wide surveys of Eucalyptus plantations for Cryphonectria canker, cankers distinctly different from those usually associated with Cryphonectria cubensis were observed. These cankers were less severe and were exemplified by cracked and slightly swollen areas on the bark. Endothia gyrosa , a well-known pathogen of woody plants including Eucalyptus spp., was consistently associated with these cankers. The pathogen is easily distinguished from C. cubensis by the presence of orange-brown stromata and non-septate ascospores. Inoculations on Eucalyptus grandis resulted in lesions similar to those observed on naturally infected trees. The disease associated with E. gyrosa is widespread in South Africa, and research is required to establish control strategies.     

Mixed inoculum for the biological control of chestnut blight1

The vegetative compatibility of four hypovirulent isolates of Cryphonectria parasitica was checked with 35 virulent isolates obtained from chestnut samples collected in many regions of Italy. Artificial inoculation tests confirmed the effectiveness of a mixture prepared with hypovirulent isolates. This was packed in squeezing tubes, to simplify the biological control of chestnut blight in practice.     

Colletotrichum species associated with chili anthracnose in Australia

Phylogenetic relationships were determined for 45 Colletotrichum isolates causing anthracnose disease of chili in Queensland, Australia. Initial screening based on morphology, ITS and TUB2 genes resulted in a subset of 21 isolates being chosen for further taxonomic study. Isolates in the C. acutatum complex were analysed using partial sequences of six gene regions (ITS, GAPDH, ACT, CHS‐1, TUB2 and HIS3), and in the C. gloeosporioides complex were analysed using four gene regions (ITS, TUB2, ApMat and GS). Phylogenetic analysis delineated four Colletotrichum species including C. siamense, C. simmondsii, C. queenslandicum, C. truncatum and a new Colletotrichum species, described here as C. cairnsense sp. nov. This is the first reported association of C. queenslandicum, C. simmondsii and C. siamense with chili anthracnose in Australia; these species were previously associated with anthracnose on papaya and avocado. Furthermore, the dominant species causing anthracnose of chili in Southeast Asia, C. scovillei, was not detected in Australia. Inoculations on chili fruit confirmed the pathogenicity of C. cairnsense and the other four species in the development of chili anthracnose in Australia.     

Horizontal transmission of hypoviruses between vegetative compatibility types of <Emphasis Type="Italic">Cryphonectria parasitica</Emphasis> in Macedonia

Biological control of chestnut blight with hypovirulence depends on the successful transmission of hypoviruses between individuals of the chestnut blight fungus, Cryphonectria parasitica. Vegetative incompatibility inhibits horizontal virus transmission, but not completely. In an effort to assess the potential for the spread of hypoviruses in the Republic of Macedonia, we studied the transmission of Cryphonectria hypovirus 1 (CHV-1) among the five observed vegetative compatibility (vc) types of C. parasitica. One fungal isolate of each vc type was infected with CHV-1 and was paired in vitro with isolates of all other vc types for a total of 20 combinations of virus donors and recipients, and 250 replicate trials per combination. Virus transmission was scored after 7 days as successful if the recipient isolate took on an unpigmented culture phenotype typical of virus infection. Transmission occurred at high frequencies between some pairs of vc types, but in <1% of the trials for 10 of the 20 combinations of donors and recipients. Asymmetric transmission was observed between some vc types that had different alleles at vegetative incompatibility loci vic1 or vic7; i.e., transmission occurred at high frequencies in one direction, but very low frequencies between the same pair of isolates in the opposite direction. The expected virus transmission, calculated as the average transmission predicted for any two randomly chosen individuals from a population, was highly negatively correlated to vc type diversity. Results for isolates of C. parasitica from Macedonia were similar to those from Italy, but less transmission was generally observed. Differences in genetic background effects on transmission may vary among different populations even when isolates differ by the same vic alleles.     

Temporal and spatial propagule deposition patterns of the emerging fungal pathogen of chestnut Gnomoniopsis castaneae in orchards of north-western Italy

Two chestnut (Castanea sativa) orchards of north-western Italy were sampled with passive spore traps 35 times over 24 months. Samples were analysed through a newly developed quantitative PCR assay to quantify propagule loads of the emerging fungal pathogen Gnomoniopsis castaneae. Average propagule deposition patterns were assessed along with temporal and climatic variables, including sampling month and season, temperatures, relative humidity, precipitations, and wind. Machine learning algorithms combining information theory, fractal analysis, unbiased recursive partitioning, ordinary least squares and logistic regressions, were used to model propagule deposition patterns. The trained models were validated on independent data gathered from 24 samplings conducted in a third chestnut orchard during the same timeframe. Results showed that propagule deposition rate (DR) was variable within and among sites, with a site average ranging from 173 to 765 spores ⋅m⁻² ⋅h⁻¹. Propagule deposition was observed across all seasons, although the DR dropped substantially during wintertime (p < 0.05). Mean, maximum, and minimum temperatures, the growing degree days at 0 and 5℃ thresholds, and wind gust were all positively correlated (p < 0.05) with DR of G. castaneae. The trained models were all significant (p < 0.05), as well as their validation (p < 0.05). Fluctuations of propagule deposition throughout the year were consistent among sites and proved to be driven by temperatures. Wind gust was associated with the overall amount of propagules deposited at site level. In future, the increase in temperatures and strong winds as a result of climate change may boost the spread of G. castaneae.     

Studies of canker and dieback of oak tree in China,with two Cytospora species described

Oak (Quercus sp.) is an excellent tree species as a windbreak, for water conservation, and for fireproofing in forests in China. However, several trees of this genus were found to be suffering from various fungal diseases. In this study, we evaluated 15 fungal pathogens that can cause dieback and canker disease in oak in China, and discovered two Cytospora species. They were identified as Cytospora quercinum sp. nov. and C. vinacea, based on detailed morphological comparisons and phylogenetic analysis of ITS, LSU, act, rpb2, tef1-α, and tub2 loci. This study is the first record of C. quercinum and C. vinacea as causal agents of dieback in oak based on pathogenicity tests conducted on 2-year-old plants in a greenhouse. In addition, this study also revealed the influence of different conditions on the growth rate of mycelia. Mycelial growth of C. quercinum and C. vinacea occurred at optimum temperatures of 20.1 and 20.8 °C, and optimum pH of 5.4 and 5.3, respectively. For these two species, utilization of glucose and fructose was highly efficient, and sucrose was the least efficient. The habitat of Quercus mongolica indicated that more attention and management are needed to prevent the occurrence of Cytospora disease in the summer in north-eastern China, and in spring and autumn in eastern and northern China. This study contributes to the understanding of the species causing canker or dieback diseases in important economic forest trees, and provides useful information for effective disease management of oak trees in China.     

Effects of fungicide on growth of Leptosphaeria maculans and L. biglobosa in relation to development of phoma stem canker on oilseed rape (Brassica napus)

Controlled‐environment and field experiments were done to investigate effects of the fungicide Punch C (flusilazole plus carbendazim) on growth of Leptosphaeria maculans and L. biglobosa in oilseed rape. In controlled‐environment experiments, for plants inoculated with L. maculans, fungicide treatment decreased lesion size and amount of L. maculans DNA in leaves; for plants inoculated with L. biglobosa, fungicide did not affect lesion size or amount of pathogen DNA. When release of ascospores was monitored using a Burkard spore sampler, the timing and pattern of ascospore release differed between the four seasons. In 2006/2007, the majority of ascospores released were L. maculans, whilst in 2007/2008 the majority were L. biglobosa; in both seasons L. maculans ascospores were released before L. biglobosa ascospores. In field experiments in 2002/2003 and 2003/2004, fungicide treatment decreased severity of stem canker on cv. Apex, but gave no significant yield response. In 2006/2007 and 2007/2008, fungicide treatment decreased phoma leaf spot incidence in autumn and stem canker severity at harvest, and increased yield. Fungicide treatment decreased stem canker severity more on cv. Courage, with a good yield response, than on cv. Canberra. In 2002/2003 and 2003/2004, fungicide treatment decreased the frequency of spread of L. maculans into stem pith tissues and in 2006/2007 fungicide decreased the amount of L. maculans DNA in stem tissues (measured by quantitative PCR). These results are used to suggest how effects of fungicides on interactions between L. maculans and L. biglobosa might affect severity of phoma stem canker and yield response.     

Characterization of hypovirulent isolates of the chestnut blight fungus, Cryphonectria parasitica from the Marmara and Black Sea regions of Turkey

Chestnut blight caused by the introduced fungus Cryphonectria parasitica has been responsible for the decline of Castanea sativa in Turkey since the 1960s. In this study, 72 C. parasitica isolates were recovered from the Marmara and Black Sea regions of Turkey showing white or cream-coloured culture morphology and were subjected to various tests to determine if they were infected by Cryphonectria hypovirus 1 (CHV-1). The vast majority of the isolates (69 out of 72) were vc type EU-1. Both mating types were found among a subsample of the isolates. The hypovirus was detected in 55 isolates by dsRNA extraction and/or virus specific RT-PCR on total RNA extracts. All but one isolates showed no or only weak phenol oxidase activity on agar medium containing tannic acid, typical of CHV-1 infected isolates. Through sequencing of a specific region of the hypovirus genome, we found that 24 hypovirus isolates belonged to the CHV-1 subtype I and six to the CHV-1 subtype F2. The distribution of the two CHV-1 subtypes in Turkey showed a clear geographic pattern. CHV-1 subtype I was only detected in the Marmara and western Black Sea region, whereas subtype F2 was restricted to the eastern part of the Black Sea region. The effectiveness of 23 hypovirulent isolates was tested against a virulent isolate on 2–3 years old chestnut sprouts. Ten hypovirulent isolates, all infected by CHV-1 subtype I, prevented canker development by more than 80 % suggesting that they might be suitable for biological control of chestnut blight in Turkey.     

High vegetative compatibility diversity of Cryphonectria parasitica infecting sweet chestnut (Castanea sativa) in Britain indicates multiple pathogen introductions

Chestnut blight, caused by Cryphonectria parasitica, was identified in Devon, UK, in December 2016. Intensive surveys detected the disease at further sites in Devon (seven), Berkshire (one), Dorset (one), Derbyshire (four) and a cluster of eight sites in southeast London. Over 570 survey samples were tested, and 227 were positive for C. parasitica by isolation and real-time PCR. A total of 227 isolates were tested for mating type, and 197 screened for vegetative compatibility group (VCG) and compared with VCGs known from mainland Europe. The same isolates were also screened for the presence of Cryphonectria hypovirus 1 (CHV-1). Eleven VCGs were identified within the UK population. Five corresponded to already known European VCGs but six were unique. The European VCGs mainly came from the Devon, Dorset, Berkshire and Derbyshire disease outbreaks, whilst unique VCGs were almost exclusively from the southeast London cluster. Both mating types were detected, but only one mating type was present at each site, with the exception of a single Devon site. Perithecia of C. parasitica were never observed at any site. CHV-1 was found in seven isolates from three different locations and was always subtype-I, which has limited hypovirulence. Therefore, although CHV-1 is associated with C. parasitica at some outbreaks, it probably has limited impact on virulence. The diversity of VCGs and their distribution at outbreak sites, together with findings of CHV-1, suggests C. parasitica has been introduced to the UK multiple times over at least two decades through international plant trade.     

Frost promotes the pathogenicity of Pseudomonas syringae pv. actinidiae in Actinidia chinensis and A. deliciosa plants

Frost occurs in all major areas of cultivation, presenting a threat for the production of kiwifruit crops worldwide. A series of experiments were performed on 1‐year‐old, potted plants or excised twigs of Actinidia chinensis and A. deliciosa to verify whether strict relationships exist between bacterial canker outbreaks from Pseudomonas syringae pv. actinidiae (Psa) attacks and the occurrence of autumn and winter frost events. The association between the occurrence of autumn frost and the sudden outbreak of bacterial canker in A. chinensis in central Italy has been confirmed. Both autumn and winter frosts promote Psa multiplication in the inoculated twigs of both species. The day after the frost, reddish exudates oozing from the inoculation sites were consistently observed in both species, and Psa was re‐isolated in some cases. During the thawing of both A. deliciosa and A. chinensis twigs, the 2‐cm upward and downward migration of Psa from the inoculation site was observed within 3 min, and the leaves were consistently colonized with the pathogen. A consistent brown discoloration, accompanied with a sour‐sap odour, was observed throughout the length of the excised twigs of both Actinidia species after Psa inoculation and winter frost. Psa inoculation induced a remarkably higher necrosis in excised twigs that were not frozen compared with P. s. pv. syringae inoculation. Antifreeze protection using irrigation sprinklers did not influence the short‐term period of Psa and P. s. pv. syringae multiplication in both A. deliciosa and A. chinensis twigs. Thus, the damage from frost, freeze thawing and the accumulation of Psa in Actinidia twigs promotes the migration of the pathogen within and between the orchards. Taken together, the results obtained in this study confirmed that A. deliciosa is more frost tolerant than A. chinensis, autumn frosts are more dangerous to these crops than winter frosts, and in the absence of Psa, young kiwifruit plants remain sensitive to frost.     

Molecular phylogenetics and microsatellite analysis reveal a new pathogenic Ceratocystis species in the Asian‐Australian clade

The ascomycete genus Ceratocystis has a broad geographic distribution and includes pathogens of a wide range of mostly woody hosts. Black rot of Colocasia esculenta (taro), a popular cultivated root crop in China, is caused by a species of Ceratocystis broadly treated as C. fimbriata sensu lato. Recently, isolates of Ceratocystis were obtained from black rot lesions on C. esculenta corms in two Chinese provinces. Sequence comparison of the ITS, partial β‐tubulin, TEF‐1α, MS204 and RPBII gene regions were used to identify these isolates and compare them to other Ceratocystis species. Furthermore, the diversity of Ceratocystis species in the Asian‐Australian clade (AAC) was investigated using 23 microsatellite markers. Results showed that the isolates from C. esculenta represent a novel species, which is morphologically and phylogenetically different to closely related species in the AAC, and is described here as Ceratocystis changhui sp. nov. In microsatellite analyses, this new species also emerged as distinct from others in the AAC. Inoculation tests showed that C. changhui sp. nov. is a virulent pathogen of C. esculenta corms, but produces only small lesions on Eucalyptus grandis and Eriobotrya japonica.