Interactions between Melamspora larici‐epitea pathotypes and the mycoparasite Sphaerellopsis filum from willow rusts

Five pathotypes of the willow rust Melamspora larici‐epitea were inoculated with 12 isolates of Sphaerellopsis filum derived from Melampsora species/forms occurring on willows. On average, 20.5% uredinial pustules produced S. filum pycnidia and rust spore production was reduced by 38.4% on leaf discs inoculated with S. filum. Some rust isolates were more readily infected by S. filum than others while some S. filum isolates caused higher levels of infections than other S. filum isolates. In general, the suppressive effects of these S. filum isolates on rust spore production were similar on the majority of rust pathotypes tested. There appeared to be a positive link between the rust pustule area and the rate of infection by S. filum. Sphaerellopsis filum inoculum densities were positively correlated with the reduction in rust spore production but not with the number of rust pustules. Implications from the results were discussed in relation to the deployment of S. filum in biological control of willow rust in willow mixture plantations which harbour more diverse rust pathotypes compared with monocultures.     

A new species of Melampsora rust on Salix elbursensis from Iran

A rust fungus was found causing stem cankers on 1‐ to 5‐year‐old stems of Salix elbursensis in the north west of Iran. The rust also forms uredinia on leaves and flowers of the host willow. Light and scanning electron microscopy revealed that the new rust is morphologically distinct from several Melampsora species occurring on the willows taxonomically close to S. elbursensis, but indistinguishable from Melampsora larici‐epitea. Examination of the internal transcribed spacer (ITS) region of the ribosomal DNA suggested that the rust fungus is phylogenetically close to Melampsora allii‐populina and Melampsora pruinosae on Populus spp. Based on both the morphological characteristics and the ITS sequence data, the rust is described as a new species –Melampsora iranica sp. nov.     

A method for estimating white pine blister rust canker age on limber pine in the central Rocky Mountains

Epidemiological studies of white pine blister rust on limber pine require a temporal component to explain variations in incidence of infection and mortality. Unfortunately, it is not known how long the pathogen has been present at various sites in the central Rocky Mountains of North America. Canker age, computed from canker length and average expansion rate, can be used to estimate infestation origin and infection frequency. To investigate relationships between canker lengths and canker ages for limber pine, we collected live white pine blister rust branch and stem cankers from three locations in Wyoming and two locations in Colorado. We quantified relationships between various measures of canker length and an estimate of canker age based on dendrochronological analysis. Total branch canker length was strongly, negatively correlated (r = ?0.79) with the first year of incomplete, annual ring formation (canker age). Mean longitudinal canker expansion rate was 8.4 cm year^?1 for branch and stem cankers where branches distal to the canker were either dead or alive. Annual longitudinal canker expansion, however, was significantly greater on a stem or branch where the portion distal to the canker was alive (11.5 cm year^?1) rather than dead (7.1 cm year^?1). For branches or stems, proximal expansion rate (i.e., toward or down stem) averaged 4.9 cm year^?1. The circumferential canker expansion rate (around branch or stem) was greater for stem cankers (8.3 cm year^?1) than for branch cankers (6.2 cm year^?1). Additional site and host tree covariates did not improve prediction of canker age. Two simple linear equations were developed to estimate a canker age from total length of a canker with the distal portion either alive or dead. An appropriate sample of canker ages can be used to determine how long a limber pine stand has been infested with white pine blister rust and how frequently infections have occurred.     

Clonal response in Salix to Melampsora rusts in short rotation coppice plantations

The appearance of Melampsora spp. rust on a range of Salix spp. clones used for short rotation coppice was recorded. The progress of the disease and its subsequent effect on leaf fall was also recorded. Rust pustules from all the clones described were examined using the Stereoscan Elec-tron Microscope. This revealed no differences in uredospore architecture which could be consis-tently related to host.     

Compatible and Incompatible Reactions of Melampsora Rust on Willow Leaves

Spore germination, growth and appresssoria formation for Melampsora willow rust were studied for compatible and incompatible hosts to elucidate penetration specificity and prehaustorial events in the infection process. Rust originating from Salix dasyclados was inoculated on a compatible (S. dasyclados) and an incompatible (Salix daphnoides) clone and, for comparison, on the non-host plants tomato, tobacco and poplar. The same experimental procedure was also carried out for rust originating from poplar. Rust development was followed for 5 days at 20 h intervals using a microscope with fluorescent equipment. The study showed that the Melampsora rust can develop on, and penetrate, the leaf regardless of whether the rust was inoculated on compatible or incompatible willow plants. However, the fungus was able to infect and reproduce itself only in compatible interactions. For willow rust in the prehaustorial stage, the study indicated that specific recognition mechanisms were unnecessary to start the infection process.     

Genetic diversity of the hyperparasite Sphaerellopsis filum on Melampsora willow rusts

The non‐specific rust hyperparasite Sphaerellopsis filum occurs naturally on Melampsora rusts of many species of the genus Salix as well as on a large range of other rust genera worldwide. To study the genetic diversity of the hyperparasitic fungus 77 S. filum isolates collected from rusts on willow clones from plantations, clone collections and natural habitats of different sites were investigated using polymerase chain reaction ‐ restriction fragment length polymorphism (PCR‐RFLP) analysis of the rDNA internal transcribed spacer regions including 5.8S rDNA and sequence analysis. Additionally, strains from Melampsora poplar rusts (4) and strains of Puccinia abrupta from Parthenium hysterophorus (5) and of P. obscura from Bellis perennis (1) were used for comparisons. Results of genetic analysis demonstrated distinct variation within the S. filum isolates. Two main groups with more than 32% difference between their nucleotide sequences were distinguished, indicating two taxa within S. filum. Within the first main group three profiles (I, II, III) were detected. The differences between these profiles were about 12%. The variation within each profile was very low (less than 2%). The second main group comprised two profiles (IV, V), which differed in 12 to 16% of their nucleotide positions. The isolates of group IV possessed a higher variation (up to 5%) within the group than those of the first main group (I, II, III). Group V was only represented by a single isolate. Neither interrelations between the S. filum profiles and the Melampsora genotypes nor a spatial distribution could be detected. It is remarkable that the six strains of S. filum from Puccinia rusts belong to one subgroup.     

Comparing Chilean and Swedish rust populations: possible impact of long‐distance transport of Melampsora rust

As willow (Salix) cultivation grows globally, worldwide monitoring of pathotypes for Melampsora rust, the most serious disease in willow, becomes increasingly important. We compared the pathotype composition of the Melampsora rust population in Chile with that in Sweden and assessed inocula exchange for the rust between the continents. For pathotyping rust isolates, a willow differential was used (a standardized set of willow test clones) to obtain clone‐virulence patterns. These patterns were used to identify virulence components involved in determining which willow clones a given individual pathogen isolate (genotype) can successfully attack. Each virulence component detected earlier in Sweden's rust population was present in Chile's rust population. Using isoenzyme analysis to classify willow species in Chile, a low genetic variation of Salix viminalis (an introduced willow species) was found, compared with the endemic species Salix humboldtiana. Comparison of rust populations for the two countries supports the present hypothesis that intercontinental inocula exchange can be a significant determinant of local pathotype structure, and consequently can be important for willow‐resistance breeding.     

Variation in leaf phenolics of field‐cultivated willow (Salix myrsinifolia) clones in relation to occurrence of Melampsora rust

Concentrations of potential antifungal phenolics in field‐cultivated willow (Salix myrsinifolia) clones were analysed during three growing seasons, and correlated to the occurrence of Melampsora rust. Consistent relationships between phenolics and rust were not found across the experimental years. There was significant clonal and temporal variation in phenolic content and rust frequency. Levels of some phenolics varied considerably within a sequence of four full‐grown leaves, but the variation in rust occurrence within the same leaf sequence was nonsignificant. The results suggest that the possible association between willow phenolics and rusts is not straightforward, and emphasize the importance of long‐term studies to investigate the chemical basis of willow rust interactions.     

Genetic relationships among genetically distinct forms of Melampsora larici‐epitea and related species based on AFLP data

Amplified Fragment Length Polymorphism (AFLP) was used to study genetic relationship among leaf rust fungi (Melampsora spp.) occurring on Salix species in short‐rotation coppice. Special interest was paid to a newly identified rust fungus found on S. viminalis in southern Sweden, morphologically similar to M. larici‐epitea but with a distinct DNA profile. Genetic distances among 40 Melampsora isolates collected from S. viminalis, S. dasyclados, S. viminalis x dasyclados, S. daphnoides, S. acutifolia and M. larici‐populina were calculated based on 101 AFLP markers. Neighbour‐joining analysis revealed the presence of six clusters, which corresponded exactly to predefined groups, namely three formae speciales of M. larici‐epitea, a stem‐infecting form of Melampsora on S. viminalis, the newly identified Melampsora on S. viminalis and a group consisting of two isolates from the poplar rust M. larici‐populina. All six clusters were well supported by bootstrap analysis (84 to 100% support). The newly identified Melampsora on S. viminalis was indicated to be genetically separated from M. larici‐epitea as well as from the stem‐infecting form of Melampsora.     

Informative value of canker morphology on the presence or absence of virus infection in chestnut blight cankers

Chestnut blight destroyed the native chestnut forests in North America and also severely affected the European chestnut trees after its introduction in the 20th century. The ascomycete fungus Cryphonectria parasitica is responsible for this serious disease and causes lethal bark cankers on susceptible chestnut trees. In Europe, however, an infection of C. parasitica with Cryphonectria hypovirus 1 (CHV‐1) causes hypovirulence in C. parasitica and reduces the severity of the disease. Hypovirulence biologically controls chestnut blight in many regions to date. In this study, our goal was to determine morphological canker characteristics that are indicative of virus presence or absence in C. parasitica. We investigated 677 chestnut blight cankers from seven different geographic locations across Europe. For each canker, we assessed canker length, stem encircling, canker depth, presence of sporulation, canker activity and virus infection. We statistically analysed the informative value of these morphological characteristics for the presence or absence of CHV‐1. However, we did not find reliable indicators. Our logistic regression analysis revealed that virus infection of C. parasitica is not clearly related to canker morphology. This implies that fungal isolations from chestnut blight cankers and assessments in the laboratory are required to determine infection with CHV‐1 unequivocally.     

Endothia gyrosa associated with severe stem cankers on plantation grown Eucalyptus nitens in Tasmania,Australia

Severe stem cankers in Eucalyptus nitens, from a 14-year-old mixed provenance plantation, were associated with infection by Endothia gyrosa, present in its teleomorph state. Surveys of incidence among canker severity classes were carried out in a thinned and pruned stand and an adjacent unthinned and unpruned stand within the affected plantation. No differences in incidence among the canker severity classes were found between the thinned/pruned and unthinned/unpruned stands or between different crown dominance classes within the unthinned/unpruned stand. However, the incidence among canker severity classes was strongly associated with bark roughness with 97% of rough-barked trees developing either annual cankers or cankers causing cambial damage. Stem cankers were found on only 11% of trees with smooth bark. Bark roughness in E. nitens was shown to differ significantly between provenances. Deployment of provenances prone to rough bark in routine plantation establishment may pose a risk of damaging stem canker outbreaks.     

Rust scorings in a plantation of Salix viminalis clones during ten consecutive years

Among pests and diseases occurring in coppiced willow plantations established to produce biofuel, Melampsora rust is the most widespread and severe. In the present study, the variation in rust attacks in a set of Salix viminalis clones was investigated. The aim was to look for trends possibly arising from changes in rust genotype composition, to get some idea about the risks of rust overcoming resistance bred into new willow varieties. Rust levels on 92 S. viminalis clones belonging to 18 different families were scored once each year during 1987–96 in a plantation in southern Sweden. Rust scorings in 1993 deviated most from the average scoring pattern of the clones. Furthermore there was a successive change, such that the longer the time interval between scorings the lower was the correlation between rust scores. Willow families were identified that deviated from the others over time. Various explanations for inconsistency in rust scores over time are discussed.     

Formation and growth of stem cankers caused by Gremmeniella abietina on young Pinus contorta

From 1990 to 1995, the formation and growth of stem cankers caused by Gremmeniella abietina on Pinus contorta var. latifolia was studied in three stands in northern Sweden. The stands were planted in 1976–80. The total number of cankers on 756 trees that were individually followed increased from 233 to 477 during the 5‐year period. With 42.0% of the cankers, the pathogen entered through or from the base of diseased branches, and 33.6% through visually undamaged bark. Most of the cankers were within 100 cm of the ground. In one of the three areas, the cankers were evenly distributed within 180 cm of the ground. The frequency of cankers facing north exceeded those facing south. The average vertical length of cankers had increased, 55.6% of cankers had increased their percentage coverage of the stem girth; 13.8% had fully girdled the stem. At two of the sites, there was a negative correlation between canker coverage of the stem circumference and tree height increment.     

Infection of Populus alba var. hickeliana by Melampsora medusae Thüm.

The first record of infection of Populus alba var. hickeliana by the American rust, Melampsora medusae Thum. is reported. The morphology of uredospores of rust on P. alba is compared with that of other Melampsora spp. infecting white poplars. The implications of these observations for the widespread establishment in the future of white × black hybrid poplars are discussed.     

Genetics of field resistance to Melampsora in Salix viminalis

The inheritance of resistance to Melampsora. spp. on Salix viminalis, the variation in resistance over space and time, and rust infection rate in relation to the geographic origin of the host was studied. The analysis showed that there was considerable additive genetic variation for rust resistance, that the results did not have high repeatability over space and time, and that the material from southernmost Sweden was most sensitive to the rusts occurring in south Sweden.     

Assessment of the effect of Nectria flute canker on wood quality within mature <Emphasis Type="Italic">Pinus radiata</Emphasis> using multiple methods

Nectria flute canker is a fungal disease that results in cankers on Pinus radiata stems. The disease, caused by the pathogen Neonectria fuckeliana, is associated with commercial pruning operations that limit knot defect to a central core to promote growth of high-quality clearwood. Nectria flute canker is present in commercial plantation forests in parts of New Zealand and Chile. Symptoms of the disease, bark cracks, stem flattenings or depressions and flute cankers, were identified on 25 mature Pinus radiata trees. Trees were felled, and discs crosscut from within, and adjacent to, the disease symptoms. The discs were photographed, and three different methods were used to determine the extent of Nectria damage beyond the defect core into the valuable clearwood sheath. These methods included a visual assessment of photographs of internal damage and two quantitative methods, based on calculations of defect extent using image processing software. Overall, flute cankers were associated with the greatest degree of encased bark and severe decay pockets, and stem flattenings the least. None of the flattened stems had internal decay. The average damage to the clearwood sheath due to flute canker was 43%, whilst that for bark cracks and flattenings was 19 and 8%, respectively. Similar results were achieved using all three methods, and the preferred method would depend on the context of the study. Using a combination of all three methods provides a robust method for measuring the damage to internal wood quality by stem pathogens and other abiotic agents.     

Distribution and frequency of Cronartium flaccidum on Melampyrum spp. in permanent sample plots in Finland

Abstract

In 2006, the distribution and frequency of pine stem rust Cronartium flaccidum was studied on Melampyrum spp. in permanent sample plots of an EU forest health monitoring program and of a BioSoil program in Finland. The frequency of Melampyrum spp. infected with C. flaccidum was low, but the rust was recorded in new locations of eastern Finland. No uredinia of C. flaccidum were observed in samples collected from over 700 plots. Telia were common on Melampyrum sylvaticum but were occasionally also found on Melampyrum pratense and Melampyrum nemorosum. Melampyrum pratense was the most common species growing in plots on mainly dry sites. Melampyrum sylvaticum was seldom detected, suggesting that alternate hosts are absent from northern Finland or that the current sample plot network is too sparse for effective disease monitoring in the north. No relationship between C. flaccidum, Melampyrum spp. and rust incidence in host trees was observed, which implies that the autoecious Peridermium pini is a more likely pathogen than C. flaccidum locally. Because plots containing M. sylvaticum occur almost solely in southern Finland, the plots with M. sylvaticum reflect the best changes in rust epidemics in that area. In the future, nutrient-rich sites containing M. sylvaticum may serve as a source of rust epidemics. It is unlikely that C. flaccidum will spread to dry sites via M. pratense.     

Pathogenic variability of various isolates of Seiridium cardinale,S. cupressi and S. unicorne inoculated on selected Cupressus clones and seedlings

Comparative studies of the pathogenicity of eight isolates of three Seiridium species were carried out by stem inoculations on Cupressus sempervirens and C. glabra seedlings and clones. Seiridium cardinale and S. cupressi caused bigger and more severe cankers than all strains of S. unicorne. During the first nine months after inoculation, the canker length of S. cardinale was larger than the cankers of S. cupressi. However, during the second year after inoculation, S. cupressi cankers were larger and more severe than those of 5. cardinale.     

The genetic composition of Peridermium pini and Cronartium flaccidum cankers on Scots pine as revealed by two multi‐allelic loci

Summary Two highly variable DNA markers were developed for Cronartium flaccidum and Peridermium pini. The markers were used to study the genetic composition of aecia of single cankers of both species. All five cankers of P. pini were homogenic and contained only one allele in both loci studied, indicating that the aecia were homozygous. The homogeneity of the cankers suggested that either genetic exchange via spermatization does not occur in autoecious P. pini populations, or that the pairing is homothallic. Two C. flaccidum cankers contained heterozygous loci. However, one canker contained genetically variable aecia indicating heterothallism, whereas the aecia of the other canker were homogenic.