Defence responses in plantation‐grown Eucalyptus globulus and Eucalyptus nitens after artificial fungal inoculation

The formation of reaction and barrier zones was studied in the xylem of Eucalyptus globulus and Eucalyptus nitens tree stems after wounding and artificial inoculation with two white rot fungi. The study had two objectives: to describe host responses in Eucalyptus spp. by light microscopy and to determine whether they would differ in a fungal treatment (wounding and inoculation by one of two fungal isolates) when compared to a control treatment (wounding only). Eucalyptus globulus and E. nitens developed similar reaction and barrier zones. The E. globulus barrier zone was characterized by kino vein formation. In both hosts, the reaction zone was primarily influenced by content and distribution of living tracheids and parenchyma cells within the sapwood. By contrast, the anatomy of the barrier zone showed similarities to the basic xylem structure of each host, except for some cell types that were newly formed (sclereids, kino veins) or increased in number (parenchyma cells, tracheids). Other cell types were reduced in number or completely absent. Host response in terms of barrier zone width appeared to be greater in the fungal than control treatment. Both wood decay fungi appeared to induce a wider barrier zone in both species than that associated with non‐specific damage caused exclusively by wounding. However, the small number of replicates available for this study was possibly insufficient to provide statistical evidence for different barrier zone width between fungal and control treatments.     

Strain‐related pathogenicity in Diplodia corticola

Diplodia corticola is one of the most aggressive fungal pathogens of Quercus species and is involved in the decline of Mediterranean cork oak forests and Californian oaks. Information regarding variation in virulence between strains is scarce. We hypothesize that D. corticola strains differ in virulence and consequently induce different symptoms in infected plants. To test this, infection assays were carried out on Quercus suber half‐sib seedlings with seven strains of D. corticola. Visual symptoms of infection (external lesions, leaf wilting, exudation and others) were recorded in parallel with physiological and biochemical parameters. All strains were able to cause lesions but at differing levels of aggressiveness. We show that internal lesion length did not correlate directly with strain aggressiveness and this agrees with physiological parameters that should be taken into account to infer about strain pathogenicity. Infection by all strains induced an overall negative impact on the net photosynthetic rate and an increase in the oxidative stress status of plants; however, significant differences were found when the effects of different strains were compared. Results also suggest that being under optimum growth conditions, prior to and during infection, allowed plants to respond to the pathogen. At the end of the experiment, some strains of D. corticola established a latent pathogen‐like relationship with cork oak. This is the first study to show that D. corticola virulence is strain‐dependent.     

Nondestructive sampling of Eucalyptus globulus and E. nitens for wood properties; II. Fibre length and coarseness

Within-tree variation in fibre length and coarseness was studied in fifty trees of E. globulus and E. nitens to develop a non-destructive sampling strategy. Trees, aged 5 to 9 years, were sampled across a range of sites in southern Australia. Simulated core samples were removed at six fixed heights easily accessible from the ground (0.5, 0.7, ... 1.5 m) and at eight percentage heights (0, 10, 20, ... 70%). Whole-tree values, calculated from percentage height data, were correlated with the core data to determine the optimal sampling height. Core samples were found to be reliable predictors of whole-tree fibre length, but results were variable for fibre coarseness. Simulated cores taken from the recommended sampling heights explained 87% and 71% of variation in whole-tree fibre length for E. globulus and E. nitens respectively and 54% and 45% of the variation in whole-tree fibre coarseness. Fibre length at all fixed heights showed good correlations with whole-tree values at all sites for E. globulus. For E. nitens the correlations were slightly lower and variable across sites. Results for fibre coarseness varied across sampling heights and sites for both species. The recommended sampling height for fibre length is 1.5 m for both species, whilst for fibre coarseness, the recommended sampling heights are 0.9 and 1.1 m for E. globulus, and 0.9 and 1.3 m for E. nitens. Radial orientation of cores was not important and neither fibre length nor coarseness were related to tree size or basic density. To estimate stand mean fibre length to an accuracy of ±5% would require sampling 9 whole trees or taking cores from 13 trees for E. globulus and 4 whole trees or cores from 8 trees for E. nitens. For estimating stand mean fibre coarseness, 10 whole trees of E. globulus and 7 whole trees are needed for E. nitens. Core sampling for stand mean coarseness would require more trees: 13 to 21 for E. globulus and 11 to 16 trees for E. nitens. Received 17 September 1998     

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.     

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.     

Genetic characterization of Botrytis cinerea isolates collected from pine and eucalyptus nurseries in Bio‐Bio Region,Chile

A total of 55 Botrytis cinerea isolates collected from Pinus radiata and Eucalyptus globulus plants cultivated in nurseries located in the Bio‐Bio Region, Chile, as well as isolates collected from native plants such as Rubus spp and Aristotelia chilensis located near the nurseries were genetically characterized. All isolates carried the Bc‐hch2 allele, thus belonging to genetic Group II, which is now referred to as B. cinerea. Genotyping based on the presence of transposons Boty and Flipper showed differences between isolates related to the plant host. Thus, transposa isolates (containing both transposons) were detected in P. radiata and E. globulus, while vacuma isolates (containing neither transposon) were detected in all plants except E. globulus. Notably, boty isolates (containing just the Boty transposon) were detected at high frequencies in all plant hosts. Analyses to detect mutations involved in resistance to fungicides such as benzimidazoles (BZ), dicarboximides and QoIs also showed differences in the studied isolates. Isolates collected from E. globulus were shown to carry mutations for all tree fungicides, while those collected from P. radiata presented mutations involved in resistance to BZ only. Isolates collected from native plant hosts did not carry any of the mutations analysed.     

Nondestructive sampling of Eucalyptus globulus and E. nitens for wood properties. I. Basic density

A non-destructive sampling strategy for basic density, based on removing 12 mm bark-to-bark cores, was developed in E. globulus and E. nitens. Fifty trees of each species, aged 5 to 9 years, were sampled across a range of sites. Core samples were removed on both a north-south and an east-west axis from 6 fixed heights in the base of the tree (0.5 m, 0.7 m, …, 1.5 m). Whole-tree values were calculated from disc samples removed at eight percentage heights (0, 10, 20, …, 70%) and correlations between the cores and whole-tree values were used to determine the optimal sampling height. Core samples were found to be reliable predictors of whole-tree density, explaining between 84% and 89% of the variation between trees. Core sampling of E. globulus and E. nitens to estimate basic density of whole-trees and stands is feasible; cores from trees at all E. globulus sites gave high correlations with whole-tree values. For E. nitens, site differences were apparent, and it is recommended that a small destructive sampling program should be undertaken prior to commencing a major sampling program. Recommended optimal sampling heights are 1.1 m for E. globulus and 0.7 m for E. nitens. Core orientation was not important and density was not related to tree size. Six whole-tree samples or eight core samples are required for estimating the mean density of a stand at a specific site to an accuracy of ±20 kg m⁻³ with a 95% confidence interval. Received 17 September 1998     

Nondestructive sampling of Eucalyptus globulus and E. nitens for wood properties. III. Predicted pulp yield using Near Infrared Reflectance Analysis

  Within-tree variation in kraft pulp yield, predicted using near infrared reflectance analysis, was studied in thirty trees of E. globulus and fifty trees of E. nitens to develop a non-destructive sampling strategy. Trees, aged 5 to 9 years, were sampled across a range of sites in southern Australia. Simulated core samples were removed at six fixed heights easily accessible from the ground (0.5, 0.7, ... 1.5 m) and at seven percentage heights (0, 20, 30, ... 70%). Whole-tree values, calculated from percentage height data, were correlated with the core data to determine the optimal sampling height. Core samples were found to be good predictors of whole-tree pulp yield for E. globulus, with simulated cores taken from the recommended sampling height (1.1 m) explaining more than 50% of variation in whole-tree pulp yield. Results for E. nitens were variable with large site differences apparent. On high quality sites, core samples from the recommended sampling height (0.9 m) were good predictors of whole-tree pulp yield, explaining around 60% of the variation. On poor quality sites, cores were poor predictors of whole-tree pulp yield. Radial orientation of cores was not important and predicted pulp yield was not related to tree size, basic density or fibre length. To estimate stand mean pulp yield to an accuracy of ±1% would require sampling 6 trees of E. globulus and 4 trees for E. nitens using either multiple discs or core samples. A single sampling height (1.1 m) is recommended for sampling for basic density, fibre length, fibre coarseness and predicted pulp yield in E. globulus. For E. nitens the recommended sampling height for basic density and fibre length is 0.7 m and 0.9 m is recommended for predicted pulp yield on good quality sites. Received 17 September 1998     

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.     

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.     

以芳香油和薪材为最终产品的桉树幼林地上部分生物量研究*

在2年生的桉树引种试验林内,对蓝桉和新引种的5种其他桉树的地上部分生物量进行了测定。研究发现,当轮伐期为2年时,不同桉树的芳香油和薪材收获量差异显著,而同一树种的芳香油和薪材产量在不同造林密度之间无显著差异。史密斯桉和贝克桉的芳香油和桉树脑含量均高于蓝桉,是生产桉树芳香油和薪材的优良树种,在我国亚热带地区有进一步试验和推广的价值。     

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.     

Analysis of differences in field performance of vegetatively and seed-propagated Eucalyptus varieties I: survival and leaf gas exchange

The aim of this study was to compare survival rates and leaf gas exchange of micro- and macro-propagated Eucalyptus grandis × E. nitens, and seed-propagated E. grandis and E. nitens, at a site in Hilton, KwaZulu-Natal, South Africa, where the conditions were suitable for cold-tolerant hybrid eucalypts. Fourteen months after planting, 50% of micropropagated E. grandis × E. nitens had survived, compared with 87% for macropropagated E. grandis × E. nitens, 53% for seed-propagated E. grandis and 93% for E. nitens. Differences in instantaneous leaf gas exchange and parameters derived from the light and CO₂responses of gas exchange were not significant between micro- and macro-propagated plants, but were significant between species. All trees used water more efficiently during photosynthesis at 16 months (winter; June 2004) than at 14 months (autumn; April 2004) after planting. However, micropropagation yielded plants that were most vulnerable to the combination of prolonged dry conditions and air frost     

Physiological responses of Eucalyptus nitens × nitens under experimentally imposed water stress

This study reports on how a cold- and frost-tolerant clone of Eucalyptus nitens × nitens responds to drought stress. The aim was to identify physiological traits that contribute to drought acclimation. Macropropagated saplings were grown in a climate-controlled greenhouse in pots filled with coarse river sand supplied with a slow-release fertiliser. One group of plants was kept regularly watered (control), and another group was subjected to four cycles of water stress (drought) whereby water was withheld for periods lasting 6, 10, 10 and 14 d, with 4 d of regular watering (recovery) inbetween. A drought cycle was terminated once saplings showed signs of wilting. Daily responses in stomatal conductance (g _s) were similar between control and drought treatments, except on the day of termination of the cycle, when g _s was significantly depressed in droughted plants. During the fourth and most severe drought cycle, there was a physiological adaptation to water stress because g _s was similar between control and droughted plants. Stomatal conductance was significantly positively correlated with volumetric soil moisture content in the drought treatment, but not in the control treatment. Electron transport capacity (J _max) increased during each drought cycle, and the increase was significant during the fourth cycle. Other parameters derived from A/c _i response curves were similar between the treatments. Under experimentally imposed water stress, E. nitens × nitens reduced leaf area, increased assimilate rate per unit leaf area, and maintained high stomatal conductance until leaves wilted. After 46 d droughted plants had accumulated half the biomass of control plants. Therefore, a cold- and frost-tolerant clone of E. nitens × nitens may be tolerant to drought stress but at a reduced growth rate because of reduced leaf area.     

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.     

Anatomical and histochemical defence responses induced in juvenile leaves of Eucalyptus globulus and Eucalyptus nitens by Mycosphaerella infection

The responses of juvenile leaves of two Eucalyptus species, with contrasting susceptibility to infection by Mycosphaerella leaf disease, were compared. The anatomical changes, accumulation of phenolics, suberin, lignin and anthocyanin and the retention of chlorophyll were studied in leaf lesions of varying developmental stages caused by species of Mycosphaerella. Enhanced resistance of Eucalyptus nitens in southern Australia was attributed to the formation of an effective lignified and suberized necrophylactic periderm, to restrict pathogen spread. Leaves of E. nitens contained isobilateral palisade which resulted in both abaxial and adaxial cell division and the initiation of a strong reinforced cellular zone from an early lesion stage. Eucalyptus globulus formed a slower, distorted necrophylactic periderm through hypertrophic changes to existing mesophyll and limited cell divisions of the single adaxial palisade layer. Deposits of lignin and suberin did not occur until later in lesion development, which were not effective in preventing further disease development. From this study it is hypothesized that tolerance of eucalypts to Mycosphaerella pathogens may be associated with constitutive mesophyll density.     

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.     

New and unexpected host associations for Diplodia sapinea in the Western Balkans

Diplodia sapinea is an important pathogen of pine trees in plantations and urban areas in many parts of the world. This pathogen has recently also been isolated from diseased Cedrus atlantica, C. deodara and Picea omorika planted as ornamentals across the Western Balkans. The aim of this study was to consider the host range of D. sapinea in Serbia and Montenegro. Diplodia sapinea was identified from a broader collection of Botryosphaeriaceae from the Western Balkans region, based on the DNA sequence data for the internal transcribed spacer (ITS) rDNA and the translation elongation factor 1α (TEF 1‐ α). The D. sapinea isolates were obtained from sixteen tree species in the genera Abies, Cedrus, Chamaecyparis, Juniperus, Picea, Pinus, Pseudotsuga and Fagus. Four species represented new hosts in the Balkans, and this is the first report of D. sapinea from F. sylvatica anywhere in the world. Pathogenicity tests were conducted on the tree hosts from which D. sapinea was isolated, as well as on P. abies, Thuja occidentalis, Prunus laurocerasus, Eucalyptus grandis and P. patula. Inoculations were made on seedlings in the field, in the greenhouse or on freshly detached branches. Inoculations on P. pungens, P. omorika, P. abies, P. menziesii, A. concolor, P. nigra and P. sylvestris resulted in death of the seedlings 5–16 weeks after inoculation. Diplodia sapinea produced lesions on J. horizontalis and P. patula seedlings and F. sylvatica cut branches. Reciprocal inoculations showed that D. sapinea is not a pine‐specific pathogen, causing disease on tree species, including those from which it had not been isolated. Not surprisingly, the pathogen was most aggressive on some species of Pinaceae.     

Mycosphaerella species occurring on Eucalyptus globulus and Eucalyptus nitens plantations of Tasmania,Australia

The genus Mycosphaerella Johanson contains many pathogens capable of causing a severe impact on the growth of susceptible eucalypt species. The lack of knowledge about which species are present in Tasmania and their potential risk to the plantation industry prompted this study into the Mycosphaerella species occurring on Eucalyptus globulus and Eucalyptus nitens plantations in Tasmania. A total of 36 plantation and five road verge sites of E. globulus and E. nitens were sampled. Five Mycosphaerella species and three species from associated anamorph genera were isolated and identified in Tasmania; Mycosphaerella nubilosa, Mycosphaerella cryptica, Mycosphaerella tasmaniensis, Mycosphaerella grandis, Mycosphaerella vespa, Coniothyrium ovatum, Sonderhenia eucalypticola and Sonderhenia eucalyptorum. The most frequently isolated species with the highest incidence and severity of infection were M. cryptica and M. nubilosa. These two species appear to have the greatest potential to damage juvenile eucalypt plantations in Tasmania. A link between Mycosphaerella vespa and Coniothyrium ovatum is described for the first time.