Aggressiveness,cultural characteristics and genetic variation of Ceratocystis fimbriata on Eucalyptus spp.

Ceratocystis wilt caused by Ceratocystis fimbriata is currently one of the most important diseases affecting Eucalyptus in Brazil. This disease is controlled by planting resistant clones; however, possible variability in the pathogen population may compromise the selection of resistant genotypes. Therefore, this study aimed to evaluate the aggressiveness of C. fimbriata isolates obtained from Eucalyptus spp, as well as their cultural characteristics and genetic variation of their ITS rDNA gene region. We found a significant isolate × clone interaction, with the isolate RM35 being the most aggressive and presenting a broader spectrum of aggressiveness, causing greater xylem discoloration on a larger number of clones. This isolate is the most suitable for artificial inoculations focusing on the selection of resistant materials. Clones CLR‐236 and CLR‐212 were identified as the most resistant and clones CLR‐223 and CLR‐240 as the most susceptible and those that are recommended as reliable comparators in artificial inoculations. All isolates were morphologically similar and differed from C. fimbriata from sweet potato by the formation of a wide mouth endoconidiophore that produces doliform endoconidia. According to the culture media and temperature applied, the most favourable conditions for mycelial growth were observed using malt extract agar (MEA) and temperatures ranging from 24 to 26°C. There was no correlation between sporulation and aggressiveness. Great variation in ITS sequences was observed, and a total of five ITS genotypes were identified among the ten isolates tested.     

Comparison of procedures to evaluate the pathogenicity of Ceratocystisfimbriata sensu lato isolates from Eucalyptus in South Africa

Ceratocystis fimbriata sensu lato (s.l.) is an important pathogen of Eucalyptus. Pathogenicity of isolates has typically been evaluated by inoculating seedlings under greenhouse conditions. It is, however, not clear how accurately this reflects pathogenicity under field conditions. In this study, five techniques to potentially screen C. fimbriata isolates for their relative pathogenicity to Eucalyptus were compared. These included: in vitro growth comparisons on artificial media; inoculations on apples; inoculation on Eucalyptus seedlings in a greenhouse; inoculations on Eucalyptus bolts freshly cut from stems of young trees; and field inoculations on young trees. Eight isolates of C. fimbriata s.l. collected from various areas in South Africa were used. There was considerable variation in growth in culture and aggressiveness of the eight isolates. Field inoculations on young trees were best correlated with inoculations of bolts (r = 0.76). Lower correlation coefficients were obtained with seedlings (r = 0.59), apple inoculations (r = 0.56), and in vitro colony growth (r = 0.42). Inoculation of bolts provides a rapid and reliable method to screen isolates of C. fimbriata s.l. for pathogenicity to Eucalyptus.     

Quambalaria eucalypti found on Eucalyptus in Indonesia

The Eucalyptus plantation industry in Indonesia has expanded rapidly during the last few decades. During routine nursery disease surveys, symptoms of a leaf and shoot blight disease were detected on Eucalyptus mother plants. Isolates were obtained from symptomatic tissues and identified using DNA sequence analyses. Phylogenetic analyses showed that the isolates were those of Quambalaria eucalypti. Pathogenicity tests were conducted with isolates of Q. eucalypti on clones of E. pellita and E. grandis × E. pellita hybrids. These resulted in symptoms similar to those observed on naturally infected plants. Eucalyptus genotypes tested showed variation in their susceptibility, highlighting the potential to select and breed for resistance and thus to manage future outbreaks of the disease. This is the first report of the pathogen in Indonesia as well as in Southeast Asia.     

Physiological responses of Eucalyptus spp. hybrids to infection by Ceratocystis fimbriata

In Brazil, the selection and planting of eucalyptus clones resistant to Ceratocystis fimbriata is the pre‐eminent control strategy for the wilt pathogen. However, little is known about host defence responses associated with resistance of eucalyptus to C. fimbriata infection. In this work, enzymatic activity, sugars, lignin, total phenols and phenolic compounds involved in the defence response of eucalyptus clones resistant (RC) and susceptible (SC) to the Ceratocystis wilt were evaluated. Changes were detected in the sugars produced by RC clones, with higher concentrations occurring compared with SC. A similar response occurred with lignin content at 28 days after inoculation (dai) in RC plants. SC plants had an increase in polyphenol oxidase and peroxidase activities at 7 dai. In contrast, RC plants demonstrated high activity of the same enzymes at 2 and 4 dai, decreasing afterwards. There was no difference in phenylalanine ammonia‐lyase activity between resistant and susceptible clones. Hydroxycinnamic acid concentration was higher in RC than in SC; however, there was no difference between RC and SC in flavonoid concentrations. RC had high concentrations catechin, chlorogenic acid and caffeic acid. Histochemical tests demonstrated the presence of phenolic compounds and lignin, at higher intensities, in xylem of RC plants. Inoculation with C. fimbriata induced defence responses in both resistant and susceptible eucalyptus clones, but both the intensity and speed of the responses were higher in RC. Alterations in the concentrations of sugars and lignin, as well as certain enzymes and phenolic compounds, could be used to predict the relative susceptibilities to Ceratocystis wilt of different phenotypes of eucalyptus clones.     

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.     

First report of bacterial wilt caused by Ralstonia solanacearum on eucalypts in South Africa

Ralstonia solanacearum, the causal agent of bacterial wilt, has one of the widest host ranges of all phytopathogenic bacteria. This pathogen was first reported on Eucalyptus spp. in the late 1980s in Brazil. Since then, there have been reports of its occurrence on this host in Australia, China and Venezuela. Early in 1997, an 18‐month‐old clonally propagated Eucalyptus grandis × Eucalyptus camaldulensis (GC) hybrid in Zululand, KwaZulu/Natal, showed signs of wilting. The vascular tissue of infected trees was dicoloured and bacterial exudation was produced from cut surfaces. The bacterium was consistently isolated from diseased tissue, purified and identified as R. solanacearum biovar 3 race 1, using the BioLog bacterial identification system. Inoculation trials were conducted on three E. grandis × E. camaldulensis clones (GC515, GC550 and GC505). Clone GC550 displayed wilting after 3 days and all cuttings subsequently died. Clones GC515 and GC505 appeared to be less susceptible with cuttings not showing signs of disease until 7 days after inoculation. After 14 days, 90 and 80%, respectively, of cuttings of these two clones had died. This is the first report of bacterial wilt on Eucalyptus in South Africa.     

The Eucalyptus stem canker pathogen Teratosphaeria gauchensis represents distinct genetic groups in Africa and South America

Teratosphaeria gauchensis (Capnodiales) causes a serious stem canker disease on commercially propagated Eucalyptus species in South America. Recently, this pathogen was detected for the first time in Africa. Very little is known regarding the biology or origin of T. gauchensis, but it has been suggested that it is native to South America. The aim of this study was to compare isolates from Africa and South America using microsatellite markers. Bayesian analysis conducted in STRUCTURE, principal coordinates analysis and a UPGMA dendrogram revealed two distinct genetic groups for these isolates. The South American isolates were more genetically diverse than those from Africa. Patterns of genetic diversity in Africa suggest that T. gauchensis could have been introduced into Zimbabwe before spreading north‐eastwards. The existence of the two genetic groups and high haplotype richness associated with the South American and Zimbabwean populations suggest that it will be more difficult to reduce the impact of disease caused by T. gauchensis in these regions than in those areas where there is limited genetic diversity.     

Spatiotemporal dynamics of bacterial wilt in Eucalyptus

Bacterial wilt (caused by Ralstonia spp.) is one of the most damaging diseases of Eucalyptus species, and is responsible for substantial losses to producers. For efficient, scientifically-based management of this disease, it is necessary to understand the various factors involved in its development, including greater knowledge of the epidemiology of the pathogen on Eucalyptus spp. This study aimed to determine the spatialtemporal dynamics of bacterial wilt in Eucalyptus under natural infection conditions. An experiment was conducted in a commercial plantation in the municipality of Itinga, state of Maranhão, Brazil, using the clone FGCA0385 (Eucalyptus urophylla var. platyphylla). The study comprised of four plots composed of 450 plants each, subdivided into nine rows with 50 plants per row with a spacing of 3 × 3 m. Disease incidence was quantified over 1 year. The spatial dynamics of the disease was determined using the dispersion index, a modified Taylor law, and the analysis of the dynamics and structure of foci. For temporal dynamics, the curve of the disease incidence progress was plotted, and the data were analysed by simple linear regression analysis fitted to three empirical models: logistic, monomolecular, and Gompertz. The distribution pattern of Eucalyptus bacterial wilt was random, as confirmed by the Index of dispersion. Analysis of the dynamics and structure of the foci, showed that 69 disease foci occurred, 44 of which were unitary, with an average number of plants per focus of 1.63. Foci had greater length in the direction of the planting line. The epidemics were best described by the monomolecular model, with an estimated incidence of Eucalyptus bacterial wilt of 27.77% in the fourth year of the study. According to the spatiotemporal dynamics of this work, management strategies such as eliminating symptomatic plants and crop renovation can be used based on economic viability.     

Spatial–temporal patterns of Ceratocystis wilt in Eucalyptus plantations in Brazil

Ceratocystis wilt, caused by Ceratocystis fimbriata, has become the most important disease in eucalyptus (Eucalyptus spp. and hybrids) plantations in Brazil. To further our understanding of the epidemiology of this disease, we surveyed eucalyptus plantations in the states of Minas Gerais and Bahia that were known to have Ceratocystis wilt or were thought to have been planted with infected rooted cuttings. There was generally higher disease incidence in the Minas Gerais plantations, which were on former Cerrado forest sites and likely had soilborne inoculum prior to planting eucalyptus. In such plantations, disease incidence was not evident before 20 months after planting but slowly increased up to 50% at 74 months. The symptomatic and killed trees were aggregated, perhaps from uneven distribution of inoculum in the soil. Also, the progression of cumulative disease incidence best fit a monomolecular model, which is typical of soilborne diseases (fixed level of initial inoculum with little or no secondary inoculum during the crop rotation). However, plots where some trees had been harvested during the rotation showed very high levels of disease incidence in the sprouts that arose from stumps, suggesting secondary spread of the pathogen on harvesting tools or machinery. Most of the Bahia plantations were on pastureland prior to eucalyptus cultivation, and the pathogen was likely introduced with infected nursery stock. In such plots, symptoms were evident as soon as 7 months after planting, and most of the mortality occurred within 12 months. The diseased trees on former pastureland sites were sometimes aggregated within planting rows, suggesting that bunches of infected nursery stock were planted together within the rows. Care should be taken in planting disease‐free planting material and spreading the pathogen on tools, but on sites with soilborne inoculum, use of resistant clones may be the only management option.     

Cryphonectria canker on Tibouchina in Colombia

Cryphonectria canker, caused by Cryphonectria cubensis, has limited the development of new Eucalyptus plantations in tropical and subtropical regions. The pathogen is commonly found on Eucalyptus, but its occurrence on other hosts in the Myrtaceae has also been documented. In this study C. cubensis is reported as the causal agent of a serious canker disease on Tibouchina spp. (Melastomataceae) in Colombia. We used morphological studies, pathogenicity tests on Eucalyptus and Tibouchina, and a phylogenetic study using partial ribosomal DNA sequence data. This is the first record of C. cubensis on a host outside the Myrtaceae.     

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.     

Biocontrol potential of Pseudomonas azotoformans,Serratia marcescens and Trichoderma virens against Fusarium wilt of Dalbergia sissoo

Wilt disease caused by Fusarium solani is a serious constraint to Dalbergia sissoo (shisham) plantations in northern India. In this study, the antagonistic potential of 40 bacterial isolates recovered from rhizophere soil of healthy shisham trees, and a well‐characterized Trichoderma species (Trichoderma virens) were tested for their possibility as biocontrol agents for F. solani. Two promising isolates (S1 and S15) were identified which inhibited pathogen growth, caused chitin degradation, produced siderophores and solubilized phosphate in vitro. Isolate S15 scored highest for hydrogen cyanide (HCN) production while isolate S1 was a non‐HCN producer. These two isolates were identified as Serratia marcescens (S1) and Pseudomonas azotoformans (S15) following sequence analysis of 16S rDNA. In dual culture assays, T. virens caused 80% inhibition of mycelial growth of the test fungus. The three selected antagonists when tested in planta in the glasshouse completely suppressed production of wilt symptoms on 12‐month‐old shisham plants. Further work is needed to ascertain the potential of these isolates to be used as biocontrol agents to manage shisham wilt under field conditions.     

Possibilités d'infection du platane par Ceratocystis fimbriata f. platani après contamination de l'eau où se développent des racines blessées

Infection of plane tree by Ceratocystis fimbriata f. platani after contamination of water as growing substrate of wounded roots. Plane tree seedlings (Platanus acerifolia) were cultivated during 1. year so as some roots emerged from the culture pot And grow in the water of a container placed below. In April, these roots were wounded and nearby, conidia suspension of C. fimbriata?. platam was injected into the water. 1 month later, foliage of some seedlings began to wilt and after 2 months, 7 out of 12 plane trees were drying. The pathogen was recovered from all parts of the roots above the water level inside the pots and as high as the collar for 4 plants.     

Population structure of Chrysoporthe austroafricana in southern Africa determined using Vegetative Compatibility Groups (VCGs)

Chrysoporthe austroafricana is one of the most damaging pathogens of Eucalyptus trees in southern Africa. It also occurs on non‐native Tibouchina granulosa trees and native Syzygium species. Additional isolates of the pathogen from previously unstudied countries in the region have become available from survey studies. The aim of this study was to use VCGs to consider the diversity in populations of isolates collected in various countries in southern Africa (Malawi, Mozambique, Namibia, South Africa and Zambia) and from different hosts. We also wanted to determine whether there are shared VCGs among these countries and hosts in southern Africa and establish a VCG tester strain database. Results showed a high diversity amongst isolates from different countries and hosts, but suggested little movement of VCGs among countries or hosts based on the available isolates. A total of 108 VCG tester strains were identified for southern Africa.     

Host range of Erwinia psidii and genetic resistance of Eucalyptus and Corymbia species to this pathogen

Dieback caused by Erwinia psidii is currently one of the most important emerging diseases in eucalypt plantations in Brazil. However, little is known in terms of the host range of this pathogen or the potential sources of resistance against the disease it causes. In this study, we inoculated plants of species from nine families to gain insight into the host range of E. psidii. Plants of all inoculated species of Myrtaceae except Acca sellowiana exhibited disease symptoms and therefore represent potential hosts for the pathogen under natural conditions. In addition, the response of four Corymbia species, 29 Eucalyptus species and three interspecific Eucalyptus hybrids to inoculation with E. psidii was evaluated. All Corymbia henryi, Corymbia maculata, Eucalyptus thozetiana, Eucalyptus cloeziana, Eucalyptus viminalis, Eucalyptus dalrympleana and Eucalyptus pilularis plants were highly resistant to the pathogen, whereas differential disease resistance was observed in the other species. This study provides important information on sources of resistance to Erwinia psidii with potential use in the development of clones with enhanced resistance in eucalypt species of economic importance.     

Factors influencing infection of Acacia mearnsii by the wilt pathogen Ceratocystis albifundus in South Africa

It is well known that species of Ceratocystis, which cause canker and wilt diseases on trees, require wounds for infection. In this regard, a number of physical and environmental factors influence the success of wound colonization by these fungi. The aim of this study was to consider the influence of wound age, stem diameter and colonization of wounds by Ophiostoma quercus on the success of infection by Ceratocystis albifundus, an important wilt pathogen of Acacia mearnsii in southern and eastern Africa, under field conditions. This was performed by doing controlled inoculations on Acacia mearnsii trees, of different diameters, with C. albifundus at different time intervals after wounding and after pre‐inoculation of wounds with O. quercus at the time of wounding. The success of infection by C. albifundus decreased significantly from 8 h after wounding and was higher on stems of greater diameter. Pre‐infection of wounds by O. quercus reduced the infection success of C. albifundus. The overall results showed that managing wounds created during forestry operations by treating wounds with naturally occurring, non‐virulent fungi, such as O. quercus, could reduce the prevalence of infections by C. albifundus.     

First report of Fusarium solani causing wilt of Melia dubia

Melia dubia, a multipurpose tree species, is gaining importance to meet the demand supply gap of timber, plywood and pulpwood . In June 2016, a serious outbreak of wilt disease was observed in M. dubia seedlings planted in the Central Nursery of Forest Research Institute (FRI), Dehradun, India. The disease led to the destruction of one hundred thousand (100,000) seedlings. Earlier in June 2012, serious wilting of M. dubia seedlings was observed in Haryana, India. The pathogen was identified as Fusarium solani following standard laboratory procedures and sequence analysis of the internal transcribed spacer (ITS) region of the ribosomal DNA (rDNA). The pathogenicity of three isolates has been proved under greenhouse conditions. This is the first report of F. solani causing wilt of M. dubia.     

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.     

First report of Calonectria cerciana causing leaf blight of Eucalyptus in northern India

Eucalyptus spp. and their hybrids are frequently cloned and mass planted across farmland tracts and commercial plantations in northern India. It is a viable feeder species to the paper and pulp industries in this region. In 2018 and 2019, during field surveys conducted in northern India, a serious leaf blight disease was frequently observed in E. tereticornis plantations. Isolation from the blighted leaf samples consistently yielded fungal isolates having Calonectria‐like morphology. Morphological features coupled with sequence analysis of partial β‐tubulin (TUB2) and partial translation elongation factor‐alpha (TEF1) gene regions of two fungal isolates confirmed the species as Ca. cerciana. In detached leaf assays and glasshouse inoculation experiments, both isolates produced symptoms similar to those observed on the naturally infected leaves. Koch's postulates were fulfilled by re‐isolating Ca. cerciana from the inoculated leaves. This work is the first to confirm that Ca. cerciana is associated with a serious leaf blight disease of Eucalyptus in northern India and is an important addition to the taxonomy of Calonectria fungi in India.     

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.