Erysiphe australiana: the cause of powdery mildew on Lagerstroemia speciosa in Brazil

Powdery mildew symptoms and signs were observed on seedlings of Lagerstroemia speciosa within an ornamental plant nursery in Viçosa, State of Minas Gerais, Brazil. Signs were most prominent as intense mycelial growth and sporulation on the shoots, new branches and leaves, which led to tissue necrosis. Based on morphological characteristics and phylogenetic analyses of ITS and 28S rDNA sequences, the powdery mildew pathogen of L. speciosa was identified as Erysiphe australiana, a well‐known powdery mildew pathogen infecting Lagerstroemia spp. worldwide. To our knowledge, this is the first report of E. australiana causing powdery mildew on L. speciosa in Brazil.     

First report of Lasiodiplodia theobromae and Pseudofusicoccum stromaticum causing dieback in Syzygium malaccense tree in Brazil

Syzygium malaccense is a woody species, known as Malay apple, which is grown as an ornamental or for fruit production in Brazil, mainly in the northern and north‐eastern regions. A high incidence of dieback has been observed in Malay apple in ornamental situations, including streets and gardens. Morphological identification, sequencing analysis of TEF+ITS gene regions and a pathogenicity test confirmed that fungi in the Botryosphaeriaceae cause the disease. This is the first report of Lasiodiplodia theobromae and Pseudofusicoccum stromaticum causing dieback on S. malaccense trees in Brazil.     

Genetic diversity of the myrtle rust pathogen (Austropuccinia psidii) in the Americas and Hawaii: Global implications for invasive threat assessments

Since the myrtle rust pathogen (Austropuccinia psidii) was first reported (as Puccinia psidii) in Brazil on guava (Psidium guajava) in 1884, it has been found infecting diverse myrtaceous species. Because A. psidii has recently spread rapidly worldwide with an extensive host range, genetic and genotypic diversities were evaluated within and among A. psidii populations in its putative native range and other areas of myrtle rust emergence in the Americas and Hawaii. Microsatellite markers revealed several unique multilocus genotypes (MLGs), which grouped isolates into nine distinct genetic clusters [C1–C9 comprising C1: from diverse hosts from Costa Rica, Jamaica, Mexico, Puerto Rico, and USA‐Hawaii, and USA‐California; C2: from eucalypts (Eucalyptus spp.) in Brazil/Uruguay and rose apple (Syzygium jambos) in Brazil; C3: from eucalypts in Brazil; C4: from diverse hosts in USA‐Florida; C5: from Java plum (Syzygium cumini) in Brazil; C6: from guava and Brazilian guava (Psidium guineense) in Brazil; C7: from pitanga (Eugenia uniflora) in Brazil; C8: from allspice (Pimenta dioica) in Jamaica and sweet flower (Myrrhinium atropurpureum) in Uruguay; C9: from jabuticaba (Myrciaria cauliflora) in Brazil]. The C1 cluster, which included a single MLG infecting diverse host in many geographic regions, and the closely related C4 cluster are considered as a “Pandemic biotype,” associated with myrtle rust emergence in Central America, the Caribbean, USA‐Florida, USA‐Hawaii, Australia, China‐Hainan, New Caledonia, Indonesia and Colombia. Based on 19 bioclimatic variables and documented occurrences of A. psidii contrasted with reduced sets of specific genetic clusters (subnetworks, considered as biotypes), maximum entropy bioclimatic modelling was used to predict geographic locations with suitable climate for A. psidii which are at risk from invasion. The genetic diversity of A. psidii throughout the Americas and Hawaii demonstrates the importance of recognizing biotypes when assessing the invasive threats posed by A. psidii around the globe.     

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.     

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.     

Molecular characterization of Ralstonia solanacearum infecting Eucalyptus spp. in Brazil

Among the bacterial pathogens of Eucalyptus in Brazil, Ralstonia solanacearum is considered one of the most important because of the characteristics of the pathogen, like the high diversity among the strains related to host range, high virulence, broad geographical distribution and its damage to the crop in recent years. Given its importance and the lack of research on this pathosystem, the present study aimed to perform a molecular characterization of different strains of infected Eucalyptus plants in Brazil. A total of 19 bacterial cultures isolated from Eucalyptus in different regions of Brazil were analysed. A 372‐bp product generated by multiplex‐PCR amplification using Nmult primers identified all the strains analysed as belonging to phylotype II. Eighteen strains were grouped into subclade IIA and one into subclade IIB. The phylogenetic tree generated from the gene sequences of endoglucanase (egl) confirmed the classification of the strains into phylotype II and separated the strains into sequevars. Strains AMC22, IBSBF2568 and IBSBF2576 were grouped into a single clade, as were strains UFV18 and UFV20, with 89% and 78% a posteriori probability, respectively, forming two new potential sequevars not yet defined. We also identified strains belonging to sequevars 41 (100% probability) and 37 (88% probability). However, most of the strains did not fit into any previously described sequevar and did not form distinct clades. The results of the analysis of fragments amplified using the ERIC‐PCR technique indicated the existence of genetic diversity among the strains studied, with a generally high correlation between similarity and the geographical origin of the strains.     

Relationships between nursery practices and field performance for Eucalyptus plantations in Brazil

The 3.5 million ha of Eucalyptus plantations in Brazil support many domestic and industrial activities. In the pulpwood sector, forest productivity has increased from 12 to 40 m³ ha^–1 year^–1 through breeding and silviculturalpractices. Nurseries have fostered thisevolution with changes in container/substratetypes, propagation methods and specificinfrastructure for each plant growth phase.Eucalyptus plants are mainly produced bycuttings, using rigid tube containers filledwith vermiculite-organic compound substrates.Stock quality is based on morphologicalcharacteristics of the plants, andenvironmental conditions are highly significantfor plant performance. Performance is dependenton physiological short-term effects related tosurvival and long-term effects of genotype/rootmorphology related to tree development. Currentsilvicultural trends are reducing establishmentcosts but increasing the risk of using lowquality plants. The development of fieldperformance models and nursery physiologicaltests is recommended for Eucalyptusplantations in Brazil. Improved experimentaldesigns are needed to capture the interactionbetween plant quality and field performance.     

Shifting cultivation effects on Brazil nut (Bertholletia excelsa) regeneration

Brazil nut (Bertholletia excelsa), has emerged as the cornerstone of the extractive economy in much of the Amazon, but the debate continues as to whether or not current harvest levels have a detrimental effect on Brazil nut seedling recruitment. While regeneration studies to date have been conducted solely within mature forest, our study provides further insight into current Brazil nut regeneration dynamics, with a unique first look at regeneration in swidden fallows within two multiple-use areas in Acre, Brazil. Recruitment of individuals ≤10 cm diameter at breast height (dbh) was evaluated within three 9 ha mature forest plots and six fallows of 0.5–1.0 ha each. Higher mean densities of B. excelsa seedlings (individuals <1.5 m in height) were observed in fallow than in mature forest (12.7 and 5.3 individuals ha⁻¹, respectively). A generalized linear mixed model analysis revealed that in addition to forest type, proximity to seed sources also explained seedling densities. Height and diameter growth of young B. excelsa individuals was positively related to light availability. Photosynthetic photon flux density levels were over three times higher in fallows versus mature forest during peak sunlight hours. Results suggest that fallow environments may provide favorable conditions for B. excelsa regeneration. Not only can anthropogenic disturbance in the form of shifting cultivation play a positive role in B. excelsa regeneration, its historic presence on the landscape could explain current Brazil nut densities and distributions. Finally, swidden fallows could potentially be managed for enhanced Brazil nut densities, which may provide an opportunity for greater income for extractive families while contributing to the sustainability of Brazil nut extraction and forest conservation in the long term.     

Cross‐infectivity of oil palm by Phytophthora spp. isolated from perennial crops in Malaysia

Bud rot disease or “Pudricion del cogollo” (PC) of oil palm is a major constraint on production in Colombia and neighbouring countries such as Brazil, Costa Rica, Ecuador, Nicaragua, Panama, Peru and Surinam. To date, there are no documented reports of Phytophthora disease of oil palm in South‐East Asia. This research, therefore, was conducted to determine the pathogenic potential of Phytophthora palmivora and Phytophthora nicotianae on oil palm using both in vitro and nursery inoculation experiments. In vitro inoculation of both P. palmivora and P. nicotianae on immature oil palm leaflets caused discoloration within 2 days of inoculation and incubation at 25 ± 1.5°C, 100% RH. Similarly, in nursery trials, lesions formed on the buds (unopened leaflets) 3 days after inoculation with P. palmivora or P. nicotianae zoospore suspensions. No lesions developed on untreated leaflets in either in vitro or nursery inoculation experiments. Phytophthora spp. were re‐isolated from leaflet lesions and confirmed as the inoculated pathogens.     

Pseudopestalotiopsis elaeidis associated with leaf spots on Cycas revoluta in Brazil

Cycas revoluta (Cycadaceae) is commonly cultivated as an ornamental tree in several countries. In April 2015, C. revoluta leaves showing brownish spots with a yellow halo that progressed to leaflet necrosis were observed in Belém, Pará state, Brazil. This study aimed to determine the aetiology of this disease on sago palm in Brazil. Based on DNA sequence-based phylogeny, morphology, and colony characterization, the fungus was identified as Pseudopestalotiopsis elaeidis. To the best of our knowledge, this is the first report of P. elaeidis associated with leaf spots on C. revoluta worldwide.     

Occurrence of leaf spot caused by Cristulariella depraedans on Acer ginnala

The occurrence of previously unknown leaf spots on Amur maples (Acer ginnala) has consistently been noticed during disease surveys in Korea since 2004. The symptoms included circular or irregular leaf spots that were greyish‐green to brownish‐grey in colour, which led to leaf senescence and premature defoliation. The morphological and cultural characteristics confirmed the identity of the causal agent as Cristulariella depraedans. This identification was supported by sequence comparisons based on ITS and LSU regions. The monospore isolate proved to be pathogenic when inoculated to the host plant under artificial conditions. This is the first report of C. depraedans infecting Amur maples in the world.     

Potential susceptibility of Australian flora to a NA2 isolate of Phytophthora ramorum and pathogen sporulation potential

Phytophthora ramorum is an invasive plant pathogen and the cause of considerable and widespread damage in nurseries, gardens and natural woodland ecosystems of the USA and Europe. It is considered to be a significant plant disease as it could cause biodiversity loss and severe economic losses in plant industries in areas where it is not yet known to exist, such as Australasia. Foliar susceptibility and sporulation potential were tested using detached‐leaf assays for 70 Australian native plant species sourced from established gardens and arboreta in California using a NA2 isolate of P. ramorum. Correa‘Sister Dawn’, Eucalyptus regnans, Isopogon cuneatus, I. formosus, Leptospermum scoparium, L. lanigerum and Melaleuca squamea were identified as potentially highly susceptible host species. Hedycarya angustifolia, Olearia argophylla, Phyllocladus aspleniifolius, Pittosporum undulatum and Podocarpus lawrencei were identified as potentially resistant. All 70 species were able to be infected with P. ramorum, as confirmed by reisolation. Putative sporulating hosts include five members of the Myrtaceae, Agonis flexuosa, Corymbia ficifolia, Eucalyptus haemastoma, E. delegatensis and E. viminalis. As a part of a precautionary strategy, the potentially highly susceptible species found in this study are suitable candidates for targeted surveillance programmes in high‐risk incursion areas of Australia and within the global horticultural trade.     

Genetic variation in the natural pollen cloud of pinus sylvestris: a study based on progeny testing

Seeds from open pollination of individual female stroboli on different days in June 1990 in a coastal clone archive of Pinus sylvestris (L.) in northern Sweden (latitude 63°39’ N) were used for a short‐term progeny experiment to study differences over time in the genetic composition of the background pollen. Analysis of progeny growth and cold acclimatization in artificial whole‐plant freezing tests suggested no genetic differences in the fertilizing pollen between pollination days. A clinal variation with the latitude of pollen origin was observed for reference progenies from fertilization with pollen of known geographical origin. Comparisons between open‐pollinated progenies and reference progenies indicate that the pollen cloud originated from approximately 1° latitude (based on height growth) or 1.5° latitude (based on freezing injury) north of the pollination site.     

Characterization of Cylindrocarpon‐like species associated with litter in the old‐growth beech forests of Central Europe

Cylindrocarpon‐like fungi are globally distributed plant pathogens and have a wide range of host species. However, very little is known about the species that live in the topsoil of forests and their potential role in reducing the natural regeneration of tree species, particularly of forest trees that produce abundant fruit only once every few years. To enhance our understanding of the species that inhabit the topsoil, we studied the diversity and pathogenicity of Cylindrocarpon‐like fungi inhabiting the litter in old‐growth mixed‐beech forests in the Carpathians (Poland) and in the Alps (Austria), and in a managed beech stand in the Krakowsko‐Cz?stochowska Highland (Poland). The fungi inhabiting the beech litter were investigated using beechnuts and pine seedlings as bait. Isolates were identified based on morphology and DNA sequencing. The pathogenicity of the most common species was investigated by inoculating beech germinants. A wide range of Cylindrocarpon‐like fungi were associated with the beech litter: 718 cultures representing 12 species were isolated. Five taxa were identified down to species level: namely Ilyonectria crassa, I. pseudodestructans, I. rufa, Neonectria candida and N. obtusispora, and seven species were identified to genus level (Neonectria or Ilyonectria species). Ilyonectria destructans, which is considered to be the sexual morph of ‘Cylindrocarpon destructans’, was not found. There were qualitative and quantitative differences between the different forest sites in terms of Neonectria and Ilyonectria species composition in beech litter. The isolation frequency and species richness of Cylindrocarpon‐like fungi were greatest in beech litter taken from old‐growth mixed‐beech forests. Neonectria and Ilyonectria species were capable of killing beech germinants, suggesting that they may play a negative role in natural beech regeneration.     

Elm yellows: A phytoplasma disease of concern in forest and landscape ecosystems

Elm yellows (EY) is a lethal or decline phytoplasma disease that affects several Ulmus (elm) species and hybrids, which is widespread in North America and Europe. The symptoms vary among the elm species. In those native to North America, main symptoms include epinasty, chlorosis, premature casting of the leaves, yellow to brown discoloration of the phloem in the roots and stem and tree death that usually occurs within 1 or 2 years from the appearance of foliar symptoms. In contrast, affected trees of European and Asian species are primarily characterized by witches’ broom as a specific symptom, do not show phloem discoloration and are less prone to decline. The disease is caused by a relatively genetically homogeneous phytoplasma, the EY agent “Candidatus Phytoplasma ulmi,” a member of the EY phytoplasma group or 16SrV group, subgroup 16SrV‐A. In nature, this pathogen exhibits a high plant host specificity. The elm leafhopper Scaphoideus luteolus is the only confirmed vector of EY phytoplasma in North America, whereas Macropsis mendax has been reported as a natural vector in Northern Italy. However, other insect vectors are likely to be involved in its natural spread. Phytoplasmas of other taxonomic groups or 16SrV subgroups, which are known to infect a wide range of plant hosts, have been identified in naturally infected elm trees. However, the pathological relevance of these “non‐elm” phytoplasmas needs to be confirmed in many cases. Their detection is mainly based on the highly sensitive nested PCR assays, while pathological data are lacking. This study summarizes, within the framework of a single comprehensive review, the current knowledge of EY. Gaps in knowledge of this disease and prospects for future research are also critically discussed.     

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.     

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.     

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.