A rapid,apple-based test for virulence in Cryphonectria cubensis isolates

The presence of the Eucalyptus canker pathogen Cryphonectria cubensis in South Africa is of concern to the local forestry industry. Quantification of the virulence of this fungus through the inoculation of trees, is both time consuming, and expensive. In this study, the potential to use apples in screening for virulence of isolates of C. cubensis, was tested using different apple cultivars and inoculation procedures. The best indication of virulence was given on Golden Delicious apples incubated at 25°C for 14 days. Here, the lesion size associated with inoculations of C. cubensis, was found to correlate significantly with the virulence of the isolates, as determined by inoculations on trees.     

Occurrence and pathogenicity of Neofusicoccum parvum and N. mangiferae on ornamental Tibouchina species

Species in the Botryosphaeriaceae are important canker pathogens of woody plants, including Eucalyptus spp. The recent discovery of the Eucalyptus pathogen, Chrysoporthe austroafricana, on ornamental Tibouchina trees raised the question as to whether Tibouchina spp. might be alternative hosts for other Eucalyptus pathogens. Therefore, the aim of this study was to consider whether species of the Botryosphaeriaceae occurring on Eucalyptus spp. might also occur on ornamental Tibouchina spp. Isolations were made from Tibouchina trees in South Africa, New Zealand and Australia. Isolates were identified using morphological characteristics and DNA‐based techniques. Neofusicoccum parvum and N. mangiferae were identified from the samples. Pathogenicity trials on T. urvilleana showed that both species are pathogenic to this host.     

Effect of environment on the response of Eucalyptus clones to inoculation with Cryphonectria cubensis

The ascomycete Cryphonectria cubensis causes severe losses in Eucalyptus plantations in South Africa and selection programmes for disease tolerance are necessary. The aim of this study was to use two C. cubensis inoculation trials, planted at different locations to assess the disease susceptibility of the clones and the effect of the environment on disease development. These two trials consisted each of 21 Eucalyptus clones (E. grandis, E. grandis × camaldulensis and E. grandis × urophylla). All trees were inoculated with a single virulent strain of C. cubensis and lesion widths measured 6 and 12 months after inoculation. Clones differed significantly in their tolerance to C. cubensis. Further, disease severity differed depending on the geographical location of the trial. A significant clone × locality (genotype × environment) interaction was observed. Therefore, screening for disease resistance should take place only in areas where clones will be commercially grown.     

Virulence of Cryphonectria cubensis on Eucalyptus species differing in resistance

Variability in virulence were detected among geographic isolates of Cryphonectria.cubensis (Bruner) Hodges. Inoculation studies of isolates, originally cultured from Brazil, Africa and Hawaii, in provenance of Eucalyptus pellita F. Muell (resistant) and E. saligna Sm. (susceptible) showed that the variation in resistance of Eucalyptus spp. is quantitatively isolate-specific; the variation in virulence quantitatively host-specific. The practical consequences of pathogenic variability of C. cubensis and the strategies of selection and breeding of Eucalyptus for resistance is discussed.     

Wound response of Eucalyptus clones after inoculation with Cryphonectria cubensis

Twenty-five different Eucalyptus grandis clones were artificially wounded and inoculated with a virulent isolate of Cryphonectria cubensis. The capacity of wounds to close through callus production was correlated with the relative susceptibility of these clones to infection by C. cubensis. Clones with the greatest capacity to close wounds were those that were also most tolerant to C. cubensis infection. Those with a lower capacity to close wounds were most susceptible to Cryphonectria canker.     

Susceptibility of Eucalyptus grandis to Cryphonectria cubensis

Artificial inoculation of rooted Eucalyptus grandis cuttings with Cryphonectria cubensis in the glasshouse showed that significant differences exist in the susceptibility of various Eucalyptus clones to the pathogen. A 61.3% difference in susceptibility was observed between the most resistant and susceptible group of clones. Results appeared, however, to have been influenced by genotype × environment interaction.     

Chrysoporthe puriensis causing canker and mortality in Pleroma mutabile in the Atlantic Forest,Brazil

Fungi of the Cryphonectriaceae family are globally known to be tree pathogens. In Brazil, several species of Chrysoporthe have been found causing stem and branch cankers in Pleroma (= Tibouchina) spp. Recently, Chrysoporthe puriensis was described as a new species causing stem and branch cankers in Pleroma granulosum, Pleroma candolleanum, and Pleroma heteromallum, all native species of the Melastomataceae family. During an investigation to collect isolates of the Cryphonectriaceae family in an important Brazilian biome, the Atlantic Forest, in Serra do Mar, structures typical of Chrysoporthe species were found in a different host, Pleroma mutabile. Fruiting bodies present in the bark of these trees were collected and isolated. The isolates were submitted on morphological characterization and phylogenetic analyses of the internal transcribed spacer (ITS) and β-tubulin gene regions using Maximum likelihood, Bayesian inference and Maximum parsimony methods. The isolates collected, for this study, were identified as C. puriensis. Pathogenicity tests on seedlings of Pleroma species and Eucalyptus clones revealed C. puriensis can infect and cause canker in these plant species as mortality. The results demonstrate the importance of delimiting the C. puriensis collection range to track its dissemination in other hosts. No host specificity was observed in the inoculation tests, suggesting this is an important finding, the pathogen causes diseases and mortality in several plants of the Atlantic Forest. Additionally, the pathogen can affect others hosts, such as Eucalyptus clones in commercial plantations.     

Lesion size induced by Chrysoporthe fungal pathogens varies between Eucalyptus species and geographic locations in Zambia

Chrysoporthe canker disease is one of the serious threats facing planted Eucalyptus in southern Africa. In order to manage this disease, planting of less susceptible species of Eucalyptus has been promoted in many countries but it is not clear which species are less susceptible for Zambian environments. The purpose of this study was to test the response of different Eucalyptus species to Chrysoporthe fungal pathogens under field conditions to identify the least susceptible species to the disease. Two-year-old Eucalyptus trees (E. grandis, E. camaldulensis, E. cloeziana and E. tereticornis) growing in low- and high-rainfall locations were inoculated separately with three Chrysoporthe fungal species (C. austroafricana, C. syzygiicola and C. zambiensis) and the induced lesion size was measured six months after inoculation. Analysis of variance revealed that lesion size differed significantly between Eucalyptus species (F_3,948 = 1 978.8, p < 0.001). Lesion size on Eucalyptus species increased in the order E. camaldulensis < E. teriticornis < E. cloeziana < E. grandis. On average, trees growing in the higher-rainfall site produced significantly larger lesions than trees growing in the low-rainfall site (F_1,948 = 565, p < 0.001), suggesting that location influences disease severity. Smaller lesions on E. camaldulensis indicate lower susceptibility to Chrysoporthe canker disease, providing an opportunity to use this species in the selection and breeding for resistant Eucalyptus genotypes for plantation development in high disease-risk sites in Zambia.     

Informative value of canker morphology on the presence or absence of virus infection in chestnut blight cankers

Chestnut blight destroyed the native chestnut forests in North America and also severely affected the European chestnut trees after its introduction in the 20th century. The ascomycete fungus Cryphonectria parasitica is responsible for this serious disease and causes lethal bark cankers on susceptible chestnut trees. In Europe, however, an infection of C. parasitica with Cryphonectria hypovirus 1 (CHV‐1) causes hypovirulence in C. parasitica and reduces the severity of the disease. Hypovirulence biologically controls chestnut blight in many regions to date. In this study, our goal was to determine morphological canker characteristics that are indicative of virus presence or absence in C. parasitica. We investigated 677 chestnut blight cankers from seven different geographic locations across Europe. For each canker, we assessed canker length, stem encircling, canker depth, presence of sporulation, canker activity and virus infection. We statistically analysed the informative value of these morphological characteristics for the presence or absence of CHV‐1. However, we did not find reliable indicators. Our logistic regression analysis revealed that virus infection of C. parasitica is not clearly related to canker morphology. This implies that fungal isolations from chestnut blight cankers and assessments in the laboratory are required to determine infection with CHV‐1 unequivocally.     

Diseases on Eucalyptus species in Zimbabwean plantations and woodlots

Zimbabwe embarked on planting Eucalyptus species in the early 1900s. Based on a robust breeding programme, it has become a major source of seed for other countries in and outside Africa. Tree health surveys conducted on Eucalyptus in some east and southern African countries over the past two decades have revealed several important fungal diseases that were previously not known in the region, but little is known regarding these problems in Zimbabwe. The aim of this study was to identify important Eucalyptus diseases across Zimbabwe's agroclimatic regions. Morphological characteristics and DNA sequence data were used to identify pathogens collected to species level. Widespread stem canker diseases, caused by species belonging to the Botryosphaeriaceae and Teratosphaeriaceae, and leaf spot diseases caused by fungi in the Capnodiales, were identified. Armillaria root and stem rot was restricted to a single site in the Eastern Highlands. Fungi that could cause canker or blue stain of timber were isolated from recently harvested stumps and included species of Ceratocystis and Ophiostoma. This study is the first to identify Eucalyptus pathogens to species level in Zimbabwe and we report for the first time the presence of the stem canker pathogen T. gauchensis in southern Africa. The results will provide a foundation for the formulation of future disease management strategies in the country.     

A serious new wilt disease of Eucalyptus caused by Ceratocystis fimbriata in Central Africa

In a recent survey of Eucalyptus clones in the Republic of Congo, Central Africa, a serious wilt and die-back disease of two different hybrid clones was observed. Affected trees ranged in age from approximately 6 months to 4 years. Isolations from symptomatic plant material consistently yielded a Ceratocystis species. On the basis of morphology and sequence data this fungus was identified as Ceratocystis fimbriata, a well-known wilt and canker pathogen of many economically important plants. The Eucalyptus isolates were compared with other Ceratocystis spp. based on sequence data generated from the ITS and 5.8S region of the rRNA operon. The results confirmed the identity of the Ceratocystis isolates from Eucalyptus as C. fimbriata and showed that they group with other C. fimbriata isolates from Brazil, South Africa and Europe. Inoculations on young Eucalyptus plants were conducted in the greenhouse and all three of the Congolese isolates tested, produced typical lesions in the bark and xylem. This study represents the first report of C. fimbriata as a pathogen of Eucalyptus in Africa. This is a serious new disease that will require considerable study in order to ensure that losses, caused by C. fimbriata, do not continue.     

Variation in virulence of Cryphonectria hypovirus 1 in Macedonia

We investigated variation in virulence of Cryphonectria hypovirus 1 (CHV‐1) to the chestnut blight fungus, Cryphonectria parasitica, in Macedonia by inoculating chestnut stems in the field. We inoculated trees with two isolates of C. parasitica, each infected with one of five isolates of CHV‐1, four of which were the same for both fungal isolates. Two virus isolates, [Sk28] and [Sk47], were significantly more virulent than the others when compared in the same fungal host isolates, as measured by reduced canker growth and increased callus formation. Mycelial growth rate in vitro was weakly correlated to canker growth or callus formation and is therefore not a reliable predictor for virulence. We found significant fungus × virus interactions for canker growth and callus formation, which seems due mainly to one virus isolate. Significant interactions were not expected because the two fungal host isolates are members of the same clone that is dominant in Macedonia and most of southeastern Europe. Phenotypic variation for response to viruses, therefore, is greater than variation revealed by the genetic markers used to define clones. More than half of the trees inoculated with virus‐free controls were dead within 2 years, and the 30% still alive after 5 years had cankers with extensive callus formation, indicating that natural virus transmission had occurred after inoculation. In contrast, only 2% of the trees inoculated with virus‐infected isolates were dead after 5 years. Hypoviruses naturally occurring in Macedonia reduce canker development and tree mortality similarly to those in other parts of southern Europe, and therefore, may have good potential for biological control of chestnut blight.     

First report of Teratosphaeria zuluensis causing stem canker of Eucalyptus grandis in Uganda

Teratosphaeria stem canker is one of the most important diseases to have emerged on non‐native plantation‐grown Eucalyptus trees globally. In 2012, Eucalyptus grandis trees with typical Teratosphaeria stem canker symptoms were observed in Uganda. Multigene sequence analyses of isolates from these cankers led to the identification of T. gauchensis, previously recorded in Uganda, and T. zuluensis. This study represents the first report of T. zuluensis in Uganda. Furthermore, this is the first report of the co‐occurrence of T. zuluensis and T. gauchensis in a single area.     

Susceptibility of Eucalyptus hybrid clones to Botryosphaeria canker in Uganda

ABSTRACT

The study assessed the susceptibility of the nine commonly grown Eucalyptus clones to Neofusicoccum species associated with Botryosphaeria canker in Uganda. The inoculation trials indicated that susceptibility of Eucalyptus hybrids differed significantly (p = .000), clones GU609, GU7, GC578, and GC796 exhibiting a higher tolerance than GC784, GC550, GU8, GC514, and GC540. The results further revealed that N. parvum was more pathogenic than N. kwambonambiense. The generated information can be exploited in sustainable forest management by expanding the growing of tolerant hybrids in areas with high Botryosphaeria canker disease pressure.     

A severe canker disease of Corymbia ficifolia caused by Quambalaria coyrecup in native and urban forests of Western Australia

A serious canker disease is affecting the health of Corymbia ficifolia in urban areas of Perth, Western Australia. Quambalaria coyrecup was frequently associated with diseased trees and its presence confirmed by morphological characters and DNA sequencing. A number of Quambalaria species have previously been identified as causal agents of canker and shoot blight diseases on a range of Eucalyptus and Corymbia species. It was therefore the aim, using glasshouse studies, to determine whether Q. coyrecup was the primary causal agent of the cankers on C. ficifolia and whether other Quambalaria species could also be associated with the disease. All seedlings inoculated with Q. coyrecup produced canker symptoms within 1 month after inoculation. Canker lesions typical of those observed in the field also occurred in the four subsequent months, after which time the trial was terminated. Inoculation with Q. cyanescens and Q. pitereka (isolated from C. ficifolia shoots) did not result in lesion development. This study establishes that Q. coyrecup is a serious pathogen of C. ficifolia and that wounding is required for canker development to occur. The management of Quambalaria canker must therefore include the minimization of artificial wounding of trees in the nursery and field.     

Incomplete movement of Cryphonectria hypovirus 1 within a vegetative compatibility type of Cryphonectria parasitica in natural cankers on grafted American chestnut trees

American chestnut trees, grafted in 1980 from large survivors, were inoculated in 1982 and 1983 with four white (European) hypovirulent strains of Cryphonectria parasitica, infected with C. hypovirus 1 (CHV1); this hypovirus has been shown to be capable of moving rapidly within the mycelium of a vegetative compatibility (vc) type of C. parasitica in blight cankers. Using a 49‐cell lattice plot, 17.8×17.8 cm, the spatial patterns and frequencies of white and pigmented isolates and white and pigmented vc types were investigated within superficial cankers on the grafts located outside the hypovirulent‐strain‐inoculated zone. Four of six cankers assayed contained white isolates, and three of the four had random spatial patterns of white isolates, based on join‐count statistics. Vc tests, using pigmented isolates and pigmented single‐spore colonies of white isolates, indicated that the majority of white and pigmented isolates recovered from each of two cankers assayed were in one vc type. White and pigmented lattice‐plot cells of the same vc type were frequently in contact with each other, indicating incomplete movement of CHV1 within a vc type. Nine and 10 vc types were found in the two cankers; it is hypothesized that small, white vc type areas in each canker may be a source of CHV1 transmission to the major vc types. Based on join‐count statistics, the spatial pattern of the single, major vc type in one canker was non‐random (aggregated), whereas the other canker had a random major vc type pattern. White and pigmented in vitro variants (sectors) of C. parasitica, that resemble white and pigmented in vivo variants in spatial contact and vc compatibility, were intermediate hypovirulent and virulent on forest American chestnuts, and dsRNA positive and negative, respectively. Incomplete movement of CHV1 within a vc type could be a major cause of the prevalence of pigmented isolates in superficial cankers on chestnut trees.     

Relationship between canker size and wood starch in American chestnut

Chestnut trees (Castanca dentata) were wounded in May and inoculated with Cryphonectria parasitica. In August, canker area was measured and the surfaces of longitudinal dissections of stem wood were stained with I₂KI to reveal the presence of starch. Sections with the most starch had smaller cankers than sections with no starch. Results show that starch reserves were related to canker size and therefore may play a role in the tree's ability to respond to infection.     

Distribution and severity of damage by Cryphonectria parasitica in the chestnut stands in Guilan province,Iran

Chestnut blight caused by Cryphonectria parasitica has recently been reported from Guilan province, the only region with natural chestnut (Castanea sativa) stands in Iran. During the past few years, chestnut stands in Iran have been seriously threatened by the fungus, incidence of the disease is increasing and vast canopies are reduced to sprouts. As yet, there is no report on the disease distribution and severity in this region. Six sites from three main growing regions of chestnut in Guilan province were selected for investigation. We report occurrence and evaluation of the damage of the disease caused by C. parasitica. To evaluate the scale of damage, the investigated trees were classified into six categories based on the degree of crown damage, the number of canker wounds and the presence of the fungus. Index of health condition was calculated for all sites. During this study, a total of 250 isolates of Cryphonectria species were obtained, of which 232 isolates were C. parasitica and 18 were Cryphonectria radicalis. Castanea parasitica was observed in all regions. Index of health condition was scored from 0 to 6 with 0 being disease free and six being the most severe infection. Index results in the investigated sites varied between 0.69–5.45 and 0.93–5.55 for years 2006 and 2007, respectively. The highest damage was found in Doran (I_H = 5.55), which is located some 100 km away from Shahbalutmahalleh, the site with lowest damage (I_H = 0.93). This is the first extended report on aspects of chestnut blight in Iran.     

The role of Spulerina simploniella in the spread of chestnut blight

The bark miner Spulerina simploniella (Lepidoptera: Gracilariidae) was found in coppice chestnut (Castanea sativa) forests in Greece but was not found in chestnut orchards. Its larvae mine under the thin periderm of young trees, 4–10 years old, while the stem bark is still smooth. Under normal conditions it does not cause any damage to the trees. However, when chestnut blight caused by Cryphonectria parasitica is present in the area, the insect may be an agent of disease spread. Experiments revealed that spraying of 23 pupation sites with a C. parasitica conidiospore suspension caused canker formation at a rate of 100% in the coppice chestnut forests of Mount Athos, North Greece. It is believed that rain during the pupation period (approximately May 23 to June 15) may deposit conidiospores on the freshly exposed phloem and cause cankers. This bark miner has been detected in several parts of Greece, however, always in intensively managed chestnut coppice forests.     

Field survey,isolation, identification and pathogenicity of Phytophthora species associated with a Mediterranean‐type tree species

Corymbia calophylla (marri), a keystone tree species in the global biodiversity hot spot of southwestern Australia, is suffering decline and mortality associated with a canker disease caused by the endemic fungus Quambalaria coyrecup. Phytophthora species are frequently isolated from the rhizosphere of C. calophylla, and a hypothesis is that Phytophthora root infection is predisposing C. calophylla to this endemic canker pathogen. Field surveys were conducted in both anthropogenically disturbed and undisturbed C. calophylla stands, from where a total of 100 rhizosphere soil samples, from both healthy and cankered trees, were collected. Phytophthora species were isolated from 26% of the samples collected, with Phytophthora incidence significantly higher on disturbed stands than in natural forests (73% and 27%, respectively). Five Phytophthora species were recovered, including P. cinnamomi, P. elongata, P. multivora, P. pseudocryptogea and P. versiformis. Under‐bark inoculations with the Phytophthora isolates caused significant lesion lengths in excised C. calophylla stems. Corymbia calophylla response to pot infestation trials in the glasshouse varied between Phytophthora species and isolates, with isolates of P. cinnamomi and P. multivora causing a significant reduction in seedling root volume and often leading to seedling death. This study demonstrates that root disease caused by Phytophthora species, especially P. cinnamomi and P. multivora, has the ability to adversely affect C. calophylla health. This study leads the way to do a dual inoculation trial with the canker pathogen Q. coyrecup, and different Phytophthora species to investigate if Phytophthora root infection predisposes C. calophylla to this canker disease.