Outbreak of shoot blight and dieback of Eucalyptus spp., caused by Pseudoplagiostoma eucalypti in Brazil

An outbreak of a new and severe disease was observed in Eucalyptus plantations of Bahia state, Brazil. An Ascomycota fungus has been frequently associated with the main symptoms of the disease namely leaf spot, branch cankers, shoot blight, defoliation, and dieback. Based on morphological characteristics, phylogenetic analysis (ITS and TEF-1α genes), and pathogenicity test on Eucalyptus plants, Pseudoplagiostoma eucalypti was identified as the causal agent of the disease. Although P. eucalytpi has been known from in Brazil since 1998, this is the first report of it causing severe disease and die-back on Eucalyptus spp. and we also record new symptoms associated with the pathogen.     

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.     

Quambalaria eucalypti a pathogen of Eucalyptus globulus newly reported in Portugal and in Europe

The Eucalyptus pathogen Quambalaria eucalypti has been reported from several subtropical and tropical countries of the Southern Hemisphere. During the course of a nationwide monitoring programme aimed at the detection of pathogens in commercial eucalypt plantations in Portugal, Q. eucalypti was found affecting Eucalyptus globulus. The identity of the pathogen was confirmed by sequence analyses of the internal transcribed spacer (ITS) region of the rDNA operon, as well as morphological characteristics. Quambalaria eucalypti is widespread throughout the eucalypt‐producing areas of Portugal. Pathogenicity studies revealed that different host genotypes showed various degrees of susceptibility to the pathogen. To our knowledge, this is the first report of Q. eucalypti on Eucalyptus spp. in Portugal and in the Northern Hemisphere.     

Identification,pathogenicity and control of powdery mildew caused by Erysiphe elevata on Eucalyptus pellita in Sumatra,Indonesia

Powdery mildews are biotrophic ascomycetes that do not typically kill their hosts, but instead interfere with normal plant growth. In Indonesia, white powdery spots were observed forming on the upper part of fully developed leaves of Eucalyptus pellita planted in sand beds of a clonal nursery. Thus, this study aimed to identify the causal agent of eucalypt powdery mildew in Indonesia as well as to develop control strategies to better manage the disease. Pathogen identification was based on morphology, molecular characterization of DNA sequencing the ITS region and pathogenicity test. In addition, considering that some powdery mildews are inhibited by free moisture on leaves, efficacy of foliar watering was tested for disease control. Phylogenetic analyses showed that isolates collected from infected eucalypt plants grouped together with other sequences of the Erysiphe elevata clade with bootstrap support of 99%; a similar result was also confirmed by morphological observations. Pathogenicity of powdery mildew on eucalypt plants was confirmed through artificial inoculation, thus fulfilling the Koch's postulates. For disease control, foliar watering reduced the incidence of powdery mildew on mother plants. The reduction of the disease incidence began to be more evident at 2 weeks after trial establishment, and in the fourth week, the incidence was below 10% in all three treatments, while the control was above 20%. Therefore, we concluded that E. elevata is the causal agent of powdery mildew in Indonesia and that one application of foliar watering per week was enough to minimize the disease impact in the nursery.     

A single dominant Ganoderma species is responsible for root rot of Acacia mangium and Eucalyptus in Sumatra

Ganoderma root rot is the most serious disease affecting commercially planted Acacia mangium in plantations in Indonesia. Numerous Ganoderma spp. have been recorded from diseased trees of this species and to a lesser extent Eucalyptus, causing confusion regarding the primary cause of the disease. In this study, a large collection of Ganoderma isolates were obtained from the roots of A. mangium showing early signs of root rot in disease centres in South Sumatra plantations. Isolates were also collected from Eucalyptus roots at Lake Toba in North Sumatra showing similar symptoms as well as from sporocarps connected to these samples. Phylogenetic analyses showed that a single Ganoderma sp., identified as G. philippii, is the major causal agent of Ganoderma root rot on A. mangium. Results from this study also showed that the isolates obtained for Eucalyptus trees in North Sumatra belong to G. philippii. Isolates from roots and connected fruiting bodies together with the morphology of the fruiting structures confirmed this identification. Symptoms associated with this pathogen are obvious and it should not be confused with other diseases. Other Ganoderma spp. found in disease centres are considered to be of minor importance and management strategies for root rot should be focused on G. philippii.     

Mycosphaerella species associated with leaf disease of Eucalyptus globulus in Ethiopia

Eucalyptus spp. are among the most widely planted exotic trees in Ethiopia. Several damaging leaf pathogens are known from Eucalyptus spp. worldwide. Of these, Mycosphaerella spp. are among the most important, causing the disease known as Mycosphaerella leaf disease (MLD). Characteristic symptoms of MLD include leaf spot, premature defoliation, shoot and twig dieback. Recent disease surveys conducted in Ethiopian Eucalyptus plantations have revealed disease symptoms similar to those caused by Mycosphaerella spp. These symptoms were restricted to E. globulus trees growing in several localities in south, south western and western Ethiopia. The aim of this study was to identify the fungi associated with this disease. This was achieved by examining ascospore germination patterns, anamorph associations and sequence data from the Internal Transcribed Spacer (ITS) region of the rRNA operon, for representative isolates. Several different ascospore germination patterns were observed, suggesting that more than one species of Mycosphaerella is responsible for MLD on E. globulus in Ethiopia. Analysis of sequence data showed that three Mycosphaerella spp., M. marksii, M. nubilosa and M. parva were present. This is the first report of these three species from Ethiopia and represents a valuable basis on which to build further studies in the region.     

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.     

Impact of Botryosphaeria,Diplodia and Neofusicoccum species on two Eucalyptus species and a hybrid: From pathogenicity to physiological performance

Species of Botryosphaeriaceae are associated with canker and dieback of Eucalyptus spp. worldwide, but little is known about their effect on the host physiology. The aim of this study was to evaluate the impact of Botryosphaeriaceae isolates from nine species in three genera (Botryosphaeria, Diplodia and Neofusicoccum), previously isolated from eucalypts, on three different Eucalyptus hosts (seedlings of E. nitens, cuttings of E. globulusand of E. globulus× E. cypellocarpa). An approach combining standard pathogenicity trials with evaluation of plant morpho‐physiological parameters was used. The size of the lesions produced revealed differences in fungal aggressiveness and host susceptibility. Isolates of Neofusicoccum kwambonambienseand Diplodia corticolawere the most aggressive, while Botryosphaeria dothidea and Diplodia seriataisolates were the least aggressive. In general, hybrid E. globulus× E. cypellocarpa plants developed smaller lesions, followed by E. nitens and E. globulus. Eucalyptus nitensplants showed minimal modifications on the morpho‐physiological profile when infected, although more severe symptoms and mortality were observed. This is probably due to a more variable genetic background of the plants. However, in general, no direct association between putative fungal aggressiveness and plant physiological disorders could be found. Results suggested that under optimal growth conditions plants manage to cope with pathogen attack and maintain their physiological performance.     

Erysiphe alphitoides causes powdery mildew on Eucalyptus gunnii

In June 2016, pot‐grown Eucalyptus gunnii plants showing typical symptoms of powdery mildew disease were found in a greenhouse in Korea. Morphological characteristics of the anamorph and molecular sequence analyses of the ITS regions and 28S rDNA verified the fungus as Erysiphe alphitoides, which is known as common and widespread powdery mildew on Quercus spp. Although E. alphitoides has been reportedly associated with powdery mildew diseases of tropical trees and shrubs other than Quercus spp., this is the first finding of E. alphitoides on a Eucalyptus tree worldwide.     

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.     

Local and systemic control of powdery mildew in eucalyptus using essential oils and decoctions from traditional Brazilian medicinal plants

Eucalyptus plants cultivated in greenhouses, nurseries and clonal minigardens are vulnerable to attack of Oidium eucalypti (powdery mildew). Essential oils and decoctions prepared from the traditional Brazilian medicinal plants Aloysia gratissima, Cordia verbenacea and Hyptis marrubioides have been shown to be effective in the control of some plant diseases. In this study, the local and systemic effects of these preparations in the control of O. eucalypti were evaluated by assessing the progress of the disease in saplings of a highly susceptible hybrid population of eucalyptus (‘urocam’). The systemic effects were also assessed by investigating the ability of the preparations to induce disease resistance, as indicated by increased peroxidase activity. The natural products were found to be as effective as a commercial fungicide (mixture of pyraclostrobin and epoxiconazole) for the control of O. eucalypti in eucalyptus plants under greenhouse conditions. Additionally, the essential oil and decoction of A. gratissima were able to exert systemic control over O. eucalypti and induce peroxidase activity in Oidium‐inoculated plants. Scanning electron microscopy revealed that the oils and decoctions from the three species exhibited direct fungitoxic activities on O. eucalypti as shown by lysis of hyphal walls and shrinking of conidia. The results presented herein suggest that oils and decoctions from A. gratissima, C. verbenacea and H. marrubioides may offer an effective, practical and ecofriendly strategy for the control of O. eucalypti in eucalyptus.     

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.     

Realized genetic gains observed in second generation seedling seed orchards of Eucalyptus pellita in Indonesia

The first generation genetic gains in four second-generation seedling seed orchards of Eucalyptus pellita established at two locations in Indonesia (Kalimantan and Sumatra) were examined by analyzing data on the height, diameter and stem form of the trees during years 1 to 3. The seed orchards each consisted of 60 open-pollinated families: 48–49 families from first-generation plus trees (improved population) and 11–12 families from natural forest (unimproved population). The realized gains for height and diameter were ca. 16 and 19%, respectively. These gains were highly significant and consistent, irrespective of age and location. However, the gains in stem form were much lower (4%) in Kalimantan than in Sumatra (21%) and only significant in the latter, possibly because of the more intensive silvicultural management regimes at the Sumatran sites. The results confirmed that the first-generation tree improvement program for E. pellita has been effective in Indonesia, and that it should provide improved seeds for operational plantation, thus raising their productivity.     

Phytophthora alticola and P. boodjera associated with decline of young Eucalyptus smithii trees in Uruguay

Eucalyptus production mainly destined to cellulose pulp production has expanded strongly in the last 30 years in Uruguay. Eucalyptus smithii has recently emerged as a promising species for cellulose pulp production. However, an average of 40% of young trees die during the first and second summer of post-planting. In this study, 32 Phytophthora isolates were obtained from 132 E. smithii young trees with root and collar rot symptoms, confirming the association of Phytophthora to E. smithii decline. Based on phylogenetic analysis of ITS, TUB2, cox1 and HSP90 gene regions and phenotypical characteristics, two species belonging to the genera Phytophthora clade 4 were identified, P. alticola (96%) and P. boodjera (4%). Tested isolates of both species significantly reduced both shoot and root dry weights of inoculated E. smithii seedlings compared to control plants. To our best knowledge, this is the first time that P. alticola and P. boodjera are recovered from young symptomatic E. smithii trees in commercial plantations as well as the first time these species are found in the Americas.     

An agglutination test for fast and specific detection of Erwinia psidii

Dieback and wilt, caused by Erwinia psidii (Ep), is one of the most important emergent diseases of Eucalyptus spp. in Brazil. Currently, pathogen detection relies on isolation of bacteria from infected plant tissue and either identification based on morphological, physiological and biochemical tests or DNA amplification using the polymerase chain reaction (PCR), which in many cases is laborious and cumbersome. Considering the need for a simpler and more rapid, yet reliable, method for detecting the pathogen, we obtained a polyclonal antibody (anti‐Ep) and developed an agglutination test for specific detection of E. psidii. The antiserum was produced against the E. psidii strain LPF534 and tested against 101 E. psidii isolates from Eucalyptus spp.; three E. psidii isolates from Psidium guajava; 23 Ralstonia solanacearum and 18 Xanthomonas axonopodis isolates pathogenic to Eucalyptus spp.; and seven endophytic isolates from Eucalyptus spp., three of which are phylogenetically related to the genus Erwinia. Results of direct ELISA indicated that a concentration as low as 3.5 µg/ml of the anti‐Ep antibody was able to detect the E. psidii antigen and that the antibody did not cross‐reacted with other bacteria pathogenic and non‐pathogenic to Eucalyptus spp. In the agglutination test, the anti‐Ep antibody showed positive reaction with all strains of E. psidii tested whereas cross‐reaction with none of the strains that belong to other taxonomic groups was observed. The agglutination test showed a detection limit of 10⁵ colony‐forming units (CFU)/ml, and its specificity was the same as that obtained by PCR amplification using E. psidii‐specific primers. These results demonstrate that the agglutination test developed here is a useful tool for specific, fast and inexpensive detection of E. psidii although only operational on pure bacterial suspensions and not yet directly from infected tissues.     

Screening of Corymbia and Eucalyptus species for resistance to Calonectria pteridis leaf blight

Calonectria leaf blight (CLB), caused by Calonectria pteridis, is a major foliar disease of eucalypt plantations in warm and high rainfall regions in Brazil. The use of resistant genotypes is the best method for disease control in the field, so identification of sources of resistance is strategic for the long‐term genetic breeding programmes of eucalypt. In this study, resistance of 13 species of Eucalyptus and 3 species of Corymbia to CLB was evaluated by spray inoculation of a spore suspension (1 × 10⁴ conidia ml^?1) of the pathogen, under controlled conditions. Eucalyptus brassiana, E. saligna, E. scias and E. agglomerata were the most resistant, while E. robusta, E. urophylla, E. camaldulensis, E. longirostrata and C. toreliana were moderately resistant and E. cloeziana, E. pellita, E. tereticornis, E. pilularis, C. maculata, E. grandis, E. dunnii and C. citriodora were the most susceptible. The broad inter and intraspecific variability of the species tested demonstrates the potential for introgression of resistance genes into valuable genotypes as a strategy for the breeding programmes and commercial plantation of eucalypt.     

Identification and pathogenicity of Alternaria alternata causing leaf spot and blight disease of Ailanthus excelsa in India

Alternaria leaf spot of Ailanthus excelsa is generally considered as minor disease in India. Recently, severe disease outbreaks were recorded in the nursery of the Forest Research Institute, Dehradun, progeny trial at Jhumpa, Haryana, and in a nearby farm field. Leaf symptoms included small circular, brown, necrotic spots with a chlorotic halo. With severe infections, leaf spots coalesced and resulted in leaf blight. A small‐spored Alternaria with concatenated conidia was isolated consistently from the leaf samples with spot symptoms. Sequence analyses of the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) and the translation elongation factor 1‐alpha (tef‐1α) gene region of two fungal isolates confirmed the species as A. alternata. In detached leaf assays, both isolates produced symptoms similar to those observed on the nursery/field‐grown plants. To validate the detached leaf assay result, pathogenicity was also demonstrated on whole plants in a glasshouse. Koch's postulates were fulfilled by re‐isolating A. alternata from the inoculated leaves. This work is the first to confirm that A. alternata is associated with leaf spot and blight disease of A. excelsa in India.     

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.     

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.     

Genetic variation in three Ceratocystis species with outcrossing,selfing and asexual reproductive strategies

Nuclear and mitochondrial markers were used to examine variation within three closely related species of tree pathogens with differing reproductive strategies. Ceratocystis eucalypti is obligately outcrossing; Ceratocystis virescens is capable of selfing due to unidirectional mating type switching; and Chalara australis is an asexual species, comprised of a single mating type. When the nuclear DNA fingerprinting markers (CAT)₅ and (CAG)₅ were used as probes against Pst I-restricted DNA, isolates of C. eucalypti were found to be highly polymorphic, and Ch. australis showed very little polymorphism. The selfing C. virescens showed an intermediate level of variation in the nuclear fingerprint markers, and much of the variation appeared to be due to differences between two forms of the species, one pathogenic to Acer and Liriodendron and another less-pathogenic form on Fagus and other hardwoods. Mitochondrial DNA (mtDNA) polymorphisms were examined by digesting total DNA with Hae III or CfoI, and C. eucalypti showed somewhat more variation in mtDNA than did C. virescens. The only polymorphism seen in the mtDNA of Ch. australis was associated with a plasmid. Selfing in C. virescens may be common and could explain an intermediate level of diversity when compared to the obligately outcrossing C. eucalypti and the asexual Ch. australis.