Investigations on Heterobasidion annosum s.lat. in central and eastern Asia with the aid of mating tests and DNA fingerprinting

To investigate the taxonomy of Heterobasidion in Eurasia, 49 specimens belonging to H. annosum sensu lato from Asia were identified with the aid of mating tests. Most of the specimens originated from north‐eastern and south‐western China and from the Altai region in southern Siberia, but a few isolates from Kirghizia, Japan and India were also tested. In addition to mating tests, the material from China was investigated with DNA fingerprinting. Heterobasidion annosum sensu stricto was identified only from the Altai region. Homokaryotic isolates from other specimens, except the Indian ones, were sexually compatible with H. parviporum, but they also showed a high degree of compatibility with H. abietinum and with the North American S group. The isolates from SW China (eastern Himalayas) mated with about equal frequency with the European strains of H. parviporum and H. abietinum. However, the DNA fingerprinting showed that these isolates were more closely related to H. parviporum, and hence they were tentatively included in this species. The North American S group was more distant from these Eurasian taxa. Four old isolates from India mated only weakly with the members of the H. annosum s.lat. According to the species concept presented, the distribution of H. parviporum extends from western Europe through southern Siberia to northern China, Japan and the eastern Himalayas. H. annosum s.str. is so far identified only from the Altai region outside Europe, and H. abietinum only from Europe.     

PCR‐RFLP markers identify three lineages of the North American and European populations of Phytophthora ramorum

Phytophthora ramorum, the cause of sudden oak death and ramorum blight, has three major clonal lineages and two mating types. Molecular tests currently available for detecting P. ramorum do not distinguish between clonal lineages and mating type is determined by cultural methods on a limited number of samples. In some molecular diagnostic tests, cross‐reaction with other closely related species such as P. hibernalis, P. foliorum or P. lateralis can occur. Regions in the mitochondrial gene Cox1 are different among P. ramorum lineages and mitochondrial genotyping of the North American and European populations seems to be sufficient to differentiate between mating types, because the EU1 lineage is mostly A1 and both NA1 and NA2 lineages are A2. In our study, we were able to identify P. ramorum isolates according to lineage using polymerase chain reaction‐restriction fragment‐length polymorphism (PCR‐RFLP) of the Cox1 gene, first by using ApoI to separate P. ramorum from other species and EU1 from North American populations, and then AvaI to distinguish between NA1 and NA2 genotypes. However, P. foliorum had the same restriction profile as P. ramorum NA1 isolates.     

Variation in Heterobasidion annosum detected by Random Amplified Polymorphic DNAs

The production of Random Amplified Polymorphic DNAs (RAPDs) by Polymerase Chain Reaction (PCR) was used to detect variation in the isolates of Heterobasidion annosum with various geographical origins. Specific RAPD products were detected for each of the intersterility groups: the European S, F, P and the North American S group. There was considerably more polymorphism found among European S than P isolates. The analysis of RAPDs was shown to be a simple and a fast way to generate DNA markers specific to the previously established intersterility groups and thus useful for diagnostic purposes.     

Extent,distribution and origin of non-native forest tree species in Europe

ABSTRACT

The management of non-native tree species in European forests has a long history, but the information on the current number and geographic distribution of these species in European forests is incomplete and scattered across various datasets. This study aims to perform an inventory of the most frequent non-native tree species growing in European forests and analyse their current extent, geographic distribution and geographic origin. Our results show that at least 150 non-native tree species are currently growing in European forests and provenance trials. The genera represented by largest number of species are Eucalyptus, Pinus, Acacia and Abies. Species growing at the largest areas are Robinia pseudoacacia (2.44 million ha), Eucalyptus globulus (1.46 million ha), Picea sitchensis (1.16 million ha) and Pseudotsuga menziesii (0.83 million ha). In total, non-native tree species in Europe are found in an area of approximately 8.54 million ha, or 4.0% of the European forest area, and the five most abundant species account for up to 77% of this area. The largest number of these 150 species were introduced from North America (71), followed by Asia (45) and Australia (20). North American species occupy by far the largest area.     

Interfertility between North American and European strains of Phlebiopsis gigantea

Thirteen homokaryotic strains of Phlebiopsis gigantea from Canada, six strains from the US and 10 strains from Europe were paired in all possible combinations in order to determine the degree of interfertility between them. The diagnosis of interfertility was based on the production of heterokaryotic fruit bodies in the pairings. Among the resulting 406 pairings, 253 (62%) fruited. Within the strains originating from Canada, USA and Europe, 64, 80 and 64% of the pairings fruited, respectively. The fruiting frequency in pairings between the Canadian and US strains was 65%, between the Canadian and European strains 55%, and between the US and European strains 67%. True hybridization between the European and North American P. gigantea was shown by analysing the single‐spore progeny using DNA fingerprinting. In spite of the relatively low interfertility in pairings within and between continents, no clear indication of the existence of intersterility groups was found. The low interfertility is probably due to the ageing of the pure cultures and to deficient fruiting ability of certain heterokaryons on agar medium. The results strongly suggest that although the North American and European strains of P. gigantea show some differentiation they can be regarded as belonging to the same biological species.     

Heterobasidion annosum group S identified in north-eastern China

Samples of Heterobasidion annosum were collected in two stands of mixed forest in Changbaishan Nature Reserve in north-eastern China. Pure cultures were isolated from 15 specimens growing on Abies, Picea, Pinus and Populus. Mating tests showed that all these collections belong to the S inter-sterility group. The compatibility of Chinese homokaryons with S homokaryons from Europe was 100%. Clamp connections appeared almost equally in both strains of each pairing, and no distinct demarcation line formed in the pairings. The mating frequency of Chinese isolates with the European F group was also high, approximately 94%. These matings, however, were usually unilateral and a more or less distinct demarcation line was present in most of the pairings. The Chinese isolates were completely intersterile with the European P group. The S group of H. annosum in north-eastern China appears to have similar morphology and ecology as in Europe. It is common on Abies and Picea, but is not a serious pathogen in the pristine forests of north-eastern China.     

Lack of genetic differentiation after host range extension argues for the generalist nature of <Emphasis Type="Italic">Pityogenes chalcographus</Emphasis> (Curculionidae: Scolytinae)

Context  

The bark beetle, Pityogenes chalcographus, is one of the main pests in Picea abies stands, and it has also been found on other European Pinaceae species. With massive introductions of North American conifer species into European forests, this insect has extended its host range to exotic Pinaceae species.     

Larval performances and life cycle completion of the Siberian moth, <Emphasis Type="Italic">Dendrolimus sibiricus</Emphasis> (Lepidoptera: Lasiocampidae), on potential host plants in Europe: a laboratory study on potted trees

The Siberian moth, Dendrolimus sibiricus, Tschtv. is the most harmful defoliator of coniferous forests in North Asia. The pest has already spread over the Urals and continues moving westwards. Recently, it has been recommended for quarantine in member countries by European and Mediterranean Plant Protection Organization (EPPO). The performances of the pest on coniferous species planted in Europe were assessed on a range of potted trees corresponding to the spectrum of economically important conifers in the EU: European larch Larix decidua, Norway spruce Picea abies, Scots pine Pinus sylvestris, European black pine Pinus nigra, and the North American species: Douglas fir Pseudotsuga menziesii and grand fir Abies grandis. Larvae showed a potential to survive and complete the development on all these host tree species. Favorable hosts were grand fir, European larch, and Douglas fir that allowed higher survival, better larval development, and as a result, yielded heavier pupae and adult moths with higher longevity. Black pine was a poor host but, however, could still support larval and pupal development. Norway spruce and Scots pine had an intermediate behavior. If accidentally introduced to Europe, the Siberian moth may become especially damaging in forest stands predominated by European larch and by the North American firs. Norway spruce and especially the two-needle pines will be less prone to intensive defoliation by this species. The fact that the pest may damage the range of economically important coniferous species should be taken into account in the pest risk assessment for Europe and also for North America where the Siberian moth occurrence is considered likely.     

Fungi dangerous at Pinus contorta with special reference to pathogens from North Europe

Examples are given of North American fungi potentially dangerous to Pinus contorta plantations in northern Europe. The pathogenicity of North European fungi is discussed. P. contorta was found to be immune or nearly immune to all European rust fungi, more resistant than P. sylvestris to Phacidium infestans and Lophodermium pinastri, but less resistant to Crumenulopsis sororia and Discella strobilina. For other fungi no such clearcut conclusions could be drawn.     

Isozyme and RAPD polymorphisms in Heterobasidion annosum in Italy

Isozyme and random amplified polymorphic DNA (RAPD) polymorphisms were used to study variability in a group of 41 isolates from the Italian population of Heterobasidion annosum. The isolates belonged to the intersterility groups P and S, and particularly to the group that is most widely distributed in Italy, group F. Isozyme analysis was effective in identifying the three intersterility groups and revealed a high degree of genetic divergence within the P group isolates; the mannose phosphate isomerase (MPI-2) locus was diagnostic in the attribution of isolates to the more correlated F and S groups. RAPDs were detected following amplification by the polymerase chain reaction (PCR). 74 RAPD fragments, obtained through amplifications with eight primers, were scored. Isolates from the 3 intersterility groups were clearly divergent based on analysis of RAPD markers. However, a similarity index calculated for the isolates within the F population indicated a high uniformity of the isolates collected throughout the Italian peninsula.     

Geographic distribution,disease symptoms and pathogenicity of Phytophthora nemorosa and Phytophthora pseudosyringae in California,USA

During the course of surveys for Phytophthora ramorum in coastal forests of California and Oregon, P. nemorosa and P. pseudosyringae were frequently isolated from foliage and stems of the same hosts as P. ramorum. Both species ranged from central California to Oregon within 50 km of the Pacific Ocean. Both were also found in the Sierra Nevada Mountains. Phytophthora nemorosa was primarily found infecting trees in coast redwood forests and was most often isolated from bay laurel leaves (Umbellularia californica), bleeding cankers on the main bole of tanoak (Lithocarpus densiflorus), and leaf and small stem tissue of redwood (Sequoia sempervirens). Phytophthora pseudosyringae was primarily isolated from hosts found in coast live oak woodlands. Bay laurel was the most common host while infection of coast live oak (Quercus agrifolia) stems was less frequent. Inoculation studies confirmed the pathogenicity of P. nemorosa and P. pseudosyringae on their most common hosts.     

Intersterility group differentiation in Heterobasidion annosum using ribosomal IGS1 region polymorphism

Summary The intergenic spacer region 1 (IGS1) of the nuclear ribosomal RNA gene repeat was examined in three European intersterility groups of the wood‐rot fungus Heterobasidion annosum. The IGS1 elements were informative for restriction fragment length polymorphism (RFLP) fingerprinting the fungal intersterility groups following digestion with two different four‐cutter restriction endonucleases. The fingerprints of isolates from the same intersterility group from different localities shared some fragments in common, but also showed specific fragments suggesting that RFLP of the IGS1 region might be used in population studies. RFLP patterns between isolates from different intersterility groups differed more markedly. RFLP were also detected in the IGS1 region of North American strains of H. annosum and in the related species Heterobasidion araucariae. The results indicate that ribosomal IGS1 region fingerprinting could be a useful tool to identify the fungal intersterility group in Heterobasidion annosum.     

Phytophthora species isolated from ash stands in Białowieża Forest nature reserve

Five Phytophthora species were isolated from the rhizosphere of ash‐dominated areas of the Bia?owieza European lowland forest nature reserve area. Morphological and DNA analyses identified Phytophthora fragariaefolia, P. plurivora, P. cactorum, P. lacustris and P. pseudosyringae. This is the first record of P. fragariaefolia in ash forests and demonstrates that several species in the potentially highly damaging Phytophthora genus are present within a near‐pristine ecosystem.     

Susceptibility of Pedicularis spp. to Cronartium ribicola and C. flaccidum in Finland

Aeciospores of Cronartium ribicola and C. flaccidum were collected from several locations in Finland and used to inoculate Pedicularis spp. and some known and suspected alternate hosts in 2008–2009. In all trials, C. ribicola formed uredinia and telia on leaves of Ribes nigrum. No uredinia or telia of C. ribicola formed on older leaves of Pedicularis spp. but both uredinia and telia were found on young leaves of P. palustris ssp. palustris. Cronartium flaccidum produced uredinia and/or telia on leaves of P. palustris ssp. palustris, P. lapponica, Vincetoxicum hirundinaria and Melampyrum sylvaticum. Neither rust infected P. sceptrum‐carolinum, Vaccinium myrtillus, Calluna vulgaris or Ledum palustre. Similar to rusts in Asia and North America, the results showed that European C. ribicola exhibit more variable host reactions and wider alternate host ranges than earlier described. Pedicularis palustris may play a role in the spread of Cronartium in natural forests.     

Fast and specific detection of the invasive forest pathogen Heterobasidion irregulare through a Loop‐mediated isothermal AMPlification (LAMP) assay

Heterobasidion irregulare is one of the most destructive fungal pathogens of pines in North America and was accidentally introduced into central Italy, where it has become invasive. The fungus is currently recommended for regulation by the European and Mediterranean Plant Protection Organisation (EPPO). In this work, an efficient diagnostic tool for the early detection of H. irregulare based on Loop‐mediated isothermal AMPlification (LAMP) coupled with two different DNA extraction methods was developed. The LAMP assay showed high specificity and good sensitivity, with a limit of detection of about 20 picograms of target DNA and time of detection of less than 40 min. The assay was successfully tested on a variety of different samples, including fungal fruiting bodies, infected plants and colonized wood. A survey on environmental samples collected in the field was also performed using the LAMP assay coupled with a rapid DNA extraction method. The possible applications of this molecular diagnostic tool encompass the monitoring of pine forests surrounding the current invasion area, laboratory or in‐field analyses of samples from suspected trees, and the surveillance in the ports of entry of wood imported from North America.     

Variation of RAMS markers within the intersterility groups of Heterobasidion annosum in Europe

Random amplified microsatellite (RAMS) markers were used for the analysis of genetic variation within and among the intersterility groups of Heterobasidion annosum. Isolates from seven European countries were used, and the analysis of 77 markers revealed banding patterns with several markers unique to intersterility (IS) groups S, P and F. The highest number of polymorphic markers and the lowest number of fixed markers was observed within IS group P. In addition, a low level of geographic differentiation was observed within IS groups S and P by amova analysis.     

香樟树上的地衣状况对杭州市空气污染梯度的指示

在过去的25年,树木上的地衣已被广泛地用作空气污染状况的指示物,并已有大量的报道,主要在欧洲和北美的温带地区使用这一技木,在其他地区(包括香港)也可广泛应用。1987年5月11日至17日,英中双方于中国林科院亚林所举办了“应用真菌学教程”培训班,在此期间,安排了这一实验。     

Leptoglossus occidentalis and Diplodia pinea: a new insect‐fungus association in Mediterranean forests

Leptoglossus occidentalis, an insect native to North America, was inadvertently introduced into Italy about 1999. The insect damages the cones of conifer trees, especially Pinus pinea (Italian Stone pine). Pinus pinea is also affected by Diplodia pinea, a fungus native to Italy, which is becoming an increasing threat because pine trees are becoming more susceptible to it as a consequence of global warming. Because the insect and the fungus both have the pine cones as a common habitat, a possible interaction between them has been postulated. The aim of this study was to ascertain whether L. occidentalis and D. pinea interact on P. pinea cones. The interaction was studied using real‐time PCR on a group of naturally infected insects collected from a forest, and a group raised in the laboratory and artificially inoculated with D. pinea conidia. Molecular analysis showed that D. pinea DNA occurred on both naturally infected and inoculated insects, but with significant differences between the two groups. The rapid and sensitive molecular technique made it possible to detect D. pinea DNA on the bodies of the insects, and to show that the native D. pinea occurred on the exotic insect.     

Hosts and distribution of the intersterility groups of Heterobasidion annosum in the highlands of Greece

The intersterility groups of 127 pure cultures of H, annosum collected from different host trees in 12 mountain areas in Greece were identified. The F group commonly caused butt rot on the fir species Abies cephalonica and Abies borisii regis. It was the only type of H. annosum found in pure fir forests. The P group caused serious root rot in pinus sylvestris stands in north-eastern Greece. In more southern mountain areas it often colonized stumps of pinus nigra but seldom killed this tree species. The S group was found in natural forests of Picea abies in northern Greece, causing butt rot of spruce. In mixed forests, the intersterility groups of H. annosum were found relatively often in stumps of tree species other than their main hosts, although some host preference seemed to occur also in stump colonization.     

Identification of Kenyan Armillaria isolates by cultural morphology,intersterility tests and analysis of isozyme profiles

Three groups of morphologically distinct isolates were observed among nine Kenyan Armillaria isolates based on their mycelium and rhizomorphs characteristics. Seven of the isolates were interfertile with testers of North American biological species III, VII and IX. However, tests with benomyl segregants BEN 433, BEN 157-10 and BEN AVK were intersterile with the same testers and also with the European A. mellea, A. lutea and A. ostoyae. The analysis of isozyme profiles showed that morphologically similar isolates had similar isozyme profiles of esterases. Their profiles however differed from those of the European A. mellea, A. lutea and A. ostoyae. On the basis of intersterility tests and analysis of isozyme profiles, the Kenyan isolates should be considered different.