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.     

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.     

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.     

Wilt and die‐back of Eucalyptus spp. caused by Erwinia psidii in Brazil

A new disease of unknown bacterial aetiology has been observed in eucalyptus stands since 2009. It is characterized by die‐back, wilting and lesions on the branches, petiole and midrib in association with macroscopic and microscopic bacterial ooze. To date, this disease has been observed in stands of clonal Eucalyptus saligna, E. grandis and E. urophylla x E. grandis hybrids and in E. dunnii seedling plantations in the states of São Paulo, Rio Grande do Sul and Mato Grosso do Sul. Considering the economic importance of eucalyptus plantations and the potential losses caused by this disease, this study aimed to identify and characterize the causal agent. Thirty‐four strains were obtained from infected plants, which were collected in the field from four locations. The inoculation of detached leaves and intact rooted cuttings supported pathogenicity in eucalyptus. The phylogenetic analysis of four housekeeping genes (16S rDNA, gapA, recA and rpoB) as well as biochemical tests confirmed the identity of strains belonging to the species Erwinia psidii. This is the first report of E. psidii as the cause of wilt and die‐back in Eucalyptus spp. in Brazil.     

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.     

Ralstonia solanacearum decreases volumetric growth of trees and yield of kraft cellulose of Eucalyptus spp.

Ralstonia solanacearum is currently one of the most important plant pathogenic bacteria worldwide, with a wide geographical distribution and host diversity. The pathogen infects more than 200 plant species belonging to approximately 50 plant families, including Eucalyptus spp. Although, high losses have been reported in nurseries, little is known on the negative impact of the disease in the field. In this study, we evaluated the incidence of R. solanacearum and its effect on volumetric growth and cellulose yield of discoloured wood chips obtained from infected trees of one clone of Eucalyptus urophylla and two hybrid clones of Eucalyptus urophylla × E. grandis. The average incidence of bacterial wilt ranged between 60.6% and 72.4%. Volumetric growth of infected trees disease decreased 78.6% and 81.7% at 18 and 30 months, respectively. The pulp screen yield of three clones decreased between 3.2 and 6.4%, with an average 4.3%. The results of this work provide useful information on the losses of volumetric growth and pulp yield of eucalypt caused by R. solanacearum.     

Detection and characterization of Ralstonia pseudosolanacearum infecting Eucalyptus sp. in Brazil

Ralstonia solanacearum sensu lato causes bacterial wilt in many agronomic crops and tree species economically important worldwide. It is a species complex that has been divided into phylotypes and sequevars, commonly related to geographic distribution. Knowledge of the phylotype composition and genetic variability in populations of this phytopathogenic bacterium is useful for implementing effective control measures. In a survey conducted in 2019, six bacterial strains were obtained from wilted Eucalyptus urophylla trees in plantations located in the municipality of Dom Eliseu, Pará state, Brazil. Multiplex PCR based on the internal transcribed spacer (ITS) indicated that the bacterial strains belonged to two different species, namely R. pseudosolanacearum (phylotype I) and R. solanacearum (phylotype II). In a phylogenetic analysis, the nucleotide sequence of the endoglucanase (egl) gene from eucalypt strains of phylotype I clustered together with sequevar 18 sequences from GenBank. Separation of the strains into two different species was confirmed by repetitive element palindromic PCR (rep‐PCR). Pathogenicity tests demonstrated that the R. solanacearum and R. pseudosolanacearum strains recovered from E. urophylla cause disease in both tomato and eucalypt plants. Until now, only R. solanacearum (Phylotype II) has been reported causing wilt symptoms on Eucalyptus spp. in Brazil. Therefore, the presence of R. pseudosolanacearum and a need for better understanding of its genetic and aggressiveness variability as well as possible differences between the two species should be considered in breeding programmes aimed at the deployment of host resistance.     

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.     

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.     

Methods of inoculation and evaluation of Erwinia psidii in eucalypt

The dieback and wilting caused by Erwinia psidii are emerging eucalypt diseases that have been observed since 2014 in the south and central‐south regions of Brazil. Field observations have shown variability in disease severity resistance among Eucalyptus spp. clones and species. It is hypothesized that this variability is due to genetic resistance. To confirm this hypothesis, inoculations in genetically distinct eucalypt plants are necessary. However, lack of an inoculation method and disease assessment makes difficult to select resistant genotypes for use in commercial plantations or genetic breeding programmes. Three inoculation methods were tested on eight clones of Eucalyptus spp. Among them, inoculum deposition with bacteria‐impregnated toothpick on the axillary buds was the simplest and most effective, capable to reproduce the disease symptoms observed under conditions of natural infection. We also developed a rating scale for disease assessment. Among eight clones tested, only Clone 1 (Eucalyptus saligna) and Clone 2 (Eucalyptus urophylla) were resistant.     

The influence of VA mycorrhizae on the wilting of Albizia procera and Dalbergia sissoo

The effect of pre-inoculation with VA mycorrhizae on Fusarium wilt of Albizia procera and Dalbergia sissoo was done in a green house trial. Inoculation with Fusarium spp. decreased plant growth compared to noninoculated controls. Mycorrhizal plants were significantly higher than nonmycorrhizal ones. In addition to plant growth, the disease severity caused by Fusarium spp. reduced significantly when inoculated with VA mycorrhizae. The lowest percentage of Fusarium spp. were found on plants when treated with VA mycorrhizae. The population of Fusarium spp. were higher in the rhizosphere of nonmycorrhizal roots. The P concentration in the leaves of mycorrhizal and nonmycorrhizal plants were higher in the absence of Fusarium spp.     

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.     

Pathogenitätstests mit Bursaphelenchus mucronatus an Kiefern und Fichtensämlingen in Deutschland

Pathogenicity tests with Bursaphelenchus mucronatus on pine and spruce seedlings in Germany Inoculation tests on Pinus sylvestris seedlings with a German, a Siberian and a French isolate of Bursaphelenchus mucronatus and on Picea abies seedlings using only the German isolate have clearly shown, under German field conditions, that the nematode can cause wilt symptoms on the apex of about 60% of the inoculated P. sylvestris plants. The nematodes in all these tests mostly remained near the inoculation site. However, further spread of the nematodes and wilt symptoms occured when a climate chamber, at 25°C, was used or when the French isolate was used outdoors. At 25°C, 10% of the inoculated plants with at least 1000 nematodes per seedling died. Seedlings' apices wilted if more than 50 nematodes per trunk were present, whereas smaller numbers of nematodes could cause partial wilting. Nevertheless, a few pine seedlings did tolerate high nematode-population densities, up to 966 per plant. No further progress of wilt of plants with symptoms was observed the following year. Inoculations of P. abies with a German isolate resulted in population development near the site of inoculation. Spruces largely tolerated nematode infestations without any development of wilt symptoms.     

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.     

Survival of Ceratocystis fagacearum in branches of trees killed by oak wilt in Minnesota

The survival of Ceratocystis fagacearum in the branches of Quercus ellipsoidalis killed by oak wilt in Minnesota depends on the time of year when the trees die. In trees, which wilt in May or June, the pathogen can be isolated from the xylem for only 1–2 months. In trees which die later in the summer it may persist through to the following spring. Observations on oak bark beetles (Pseudopityophthorus spp.) breeding in the dead trees do not suggest that these beetles are important vectors of the disease in Minnesota.     

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.     

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.     

Pestalotiopsis biciliata,a new leaf pathogen of Eucalyptus spp. recorded in Italy

The present paper describes new symptoms on Eucalyptus spp. in Central Italy associated with the fungal pathogen Pestalotiopsis biciliata. This fungal species has been previously recorded in Italy on grape associated with fruit rot. Artificial inoculation trials on leaves confirmed the pathogenicity of the fungus to Eucalyptus camaldulensis. P. biciliata is included in the list of Pestalotiopsis spp. responsible for leaf disease on eucalypts worldwide.     

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.     

Mutualistic symbiosis between Bursaphelenchus xylophilus and bacteria of the genus Pseudomonas

Interactions between the pine wood nematode (PWN), Bursaphelenchus xylophilus, and bacteria of the genus Pseudomonas were examined by cultivating axenic PWN and bacterial strains using callus of Pinus thunbergii. Ten (Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas cepacia and Pseudomonas spp.) of the 29 bacterial strains tested, significantly increased the reproduction of PWN. The rest of the bacteria (19 strains of 10 species) inhibited the reproduction of PWN completely. The growth of 18 of the 29 bacterial strains tested, including the 10 strains promoting PWN reproduction, was significantly increased by the presence of PWN. It indicated a mutualistic symbiotic relationship between PWN and the 10 bacterial strains in the genus Pseudomonas. The bacterial mutualistic symbionts are organisms, which may have co‐evolved with PWN rather than being accidentally associated. The finding provides further evidence for our hypothesis that pine wilt disease is complex, induced by both PWN and associated phytotoxin‐producing bacteria.