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1.
Occurrence and ITS diversity of wood‐associated Bursaphelenchus nematodes in Scots pine forests in Switzerland 
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下载免费PDF全文 The quarantine pathogen Bursaphelenchus xylophilus (pine wood nematode, PWN) represents a serious threat for Pinus species in Europe. To exclude its presence in Switzerland, in 2010 and 2011 a countrywide survey was conducted in 102 Pinus sylvestris stands, chosen according to whether they contained dying or dead trees or were located in areas at risk of PWN introduction. In total, 285 trees (1–5 per site) were sampled. Nematodes were extracted from wood chips using a standard procedure, and identified to species by internal transcribed spacer (ITS) sequencing. Bursaphelenchus species were present in 34% of the trees, but no B. xylophilus was identified, i.e. PWN is still not present in Switzerland. The nematodes found belonged to seven different species, with B. vallesianus the most frequent species, followed by B. sexdentati, B. mucronatus kolymensis and B. eggersi. Three other species (B. borealis, B. pinophilus, B. poligraphi) were each only present in one or two trees. Three groups of sequences could not be assigned to a species because of the lack of matching reference sequences. The species composition found in Switzerland suggests co‐existence of southern and central European Bursaphelenchus species. Intraspecific ITS variability differed considerably among the four most common species. Bursaphelenchus eggersi, B. mucronatus kolymensis and B. sexdentati had several variable sites in the ITS region, resulting in multiple ITS genotypes in each species. In contrast, all 99 B. vallesianus isolates had an identical ITS region. This could indicate a founder effect, and possibly that B. vallesianus is not native to the Alpine region. 相似文献
2.
An overview of the occurrence of species of the genus Bursaphelenchus in Romania is presented. The data is based on recent surveys conducted for the first time throughout the country, to monitor and evaluate the potential entry of the pine wood nematode, Bursaphelenchus xylophilus. Wood samples were collected from declining trees, wood‐processing companies and imported wood packaging material. Of the 895 wood samples examined, 11 contained Bursaphelenchus specimens. Morphological and molecular analyses were carried out to characterize the species detected. With respect to the possible presence of B. xylophilus, all samples were negative, confirming the absence of this quarantine pest in Romania. Nevertheless, five Bursaphelenchus species were found: B. hofmanni, B. poligraphi, B. vallesianus, B. willibaldi, and one putative new species belonging to the sexdentati group, classified here as Bursaphelenchus sp. NR512. These results constitute the first report of the genus Bursaphelenchus in Romania. 相似文献
3.
Lin Huang Xu-ling Xu Xiao-qin Wu Feng-mao Chen Chao Li Jian-ren Ye 《Phytoparasitica》2010,38(4):369-377
The pine wood nematode (PWN), Bursaphelenchus xylophilus, is a disastrous pathogen of the pine forests in East Asia and Europe. Plant quarantine is one of the most important ways
to prevent its infection in current situation. A nested polymerase chain reaction (PCR) assay targeting the topoisomerse I
gene has been developed to detect PWN in this study. To assess the specificity of the assay, 44 morphologically characterized
nematode isolates including B. xylophilus, B. mucronatus, B. hofmanni, Seinura wuae, S. lii and Aphelenchoides macronucleatus were tested. Positive reactions characterized by amplification product of 509 bp were shown from all isolates of PWN. The
nested PCR assay can detect 50 femtogram (fg) of template DNA or one individual nematode, as small as an egg. The validity
was evaluated by analyzing the nematode samples extracted from the nematode-infested wood in the field. These results show
that the assay is a specific, sensitive method for detection of PWN with the potential in relation to the pest risk assessment
and quarantine regulations. 相似文献
4.
Official analyses undertaken in the framework of an official survey, or import controls need reliable results. This can be achieved by using validated methods. For morphological tests, this validation process is rarely illustrated despite guidance provided in EPPO PM 7/98 (1) Specific requirements for laboratories preparing accreditation for a plant pest diagnostic activity. This paper presents validation for the morphological identification of Bursaphelenchus xylophilus, with the evaluation results of published identification keys and of internally designed identification keys at B. xylophilus group and species levels. For published identification keys some criteria were shown not to be reliable for routine use: excretory pore position and number of caudal papillae. The key designed in the laboratory for xylophilus group identification was shown to be sensitive and specific when one male and one female were observed. The key designed for B. xylophilus species identification is sensitive, specific and reproducible if only one female of B. xylophilus is observed. The tools designed were validated as simple and reliable for routine analysis. The advantages and limitations of the validation process for morphological tools are discussed for process improvement. 相似文献
5.
Improved morphological key to the species of the xylophilus group of the genus Bursaphelenchus Fuchs, 1937 下载免费PDF全文
下载免费PDF全文 Correct identification of the pine wood nematode Bursaphelenchus xylophilus becomes more difficult, as more related or similar species are described. Spread of the nematode is being monitored worldwide, and in view of the fact that several new Bursaphelenchus species have been described, identification keys should be adjusted accordingly. Based on the identification key of the xylophilus group published by Braasch (2008) and the revised intrageneric grouping of Bursaphelenchus by Braasch et al. (2009), an improved dichotome morphological key including newly described species (B. paraluxuriosae, B. populi, B. macromucronatus, B. firmae, B. koreanus and B. gillanii) is presented. The recognition of the quarantine pest B. xylophilus using morphological characteristics in laboratories of the National Plant Protection Services is facilitated by a simplified key. 相似文献
6.
M. B halov 《EPPO Bulletin》2006,36(3):467-469
The State Phytosanitary Administration of the Czech Republic (SPA) conducted a survey of Bursaphelenchus xylophilus in 2004 and 2005. In 2004 97 locations were surveyed and 60 samples were taken. B. xylophilus was not found in any of the samples. In 2005, the SPA performed 120 phytosanitary inspections on solitary conifers, and in forest stands and clumps of trees. Checks were performed on Pinus sylvestris and Pinus nigra trees and in coniferous woodland mainly in areas at risk of invasion from B. xylophilus. Of 70 phytosanitary inspections performed in at risk areas, 13 samples were taken for laboratory processing and analysis. Fifty phytosanitary inspections were carried out outside risk areas, mainly in forests and parks, and 28 samples were taken. The Czech University of Agriculture in Prague together with the diagnostic laboratory of the SPA carried out a second survey in 2005. Fifty‐eight samples were taken from 58 surveyed locations in forests where coniferous trees with symptoms had been found. Laboratories performed a survey of B. xylophilus vectors to find only one vector species, Monochamus galloprovincialis subsp. pistor. In total in 2005, 178 phytosanitary inspections were carried out in the Czech Republic. B. xylophilus was not detected in either samples or vectors. Thus the results of the surveying were negative in both 2004 and 2005. The status of Bursaphelenchus xylophilus in the Czech Republic is: absent, confirmed by survey. 相似文献
7.
The Bursaphelenchus genus (Nematoda: Parasitaphelenchidae) comprises mostly wood‐inhabiting nematodes that feed on various tree‐colonizing fungi. One species of the genus, B. xylophilus, has been proven as an agent causing pine wilt disease (PWD). However, involvement of other Bursaphelenchus species in the PWD remains enigmatic. In the current paper, comparative molecular analysis is performed based on nuclear ribosomal DNA (rDNA) of B. vallesianus, a species that was recently isolated from pine trees (Pinus sylvestris) exhibiting wilting and declining symptoms in the Czech Republic. Sequencing of the nuclear‐encoded ITS1–5·8S–ITS2 rDNA region confirmed previous taxonomic conclusions based on morphology. Evolutionary reconstructions resulted in a phylogenetic tree, where the Czech isolate of B. vallesianus occupied a common clade together with other species belonging to the so‐called B. sexdentati group. Unexpectedly, comprehensive analysis of the sequence data revealed a genetic variation distinguishing the Czech isolate of B. vallesianus from all other species of the B. sexdentati group. This dissimilarity consists of the presence of a four nucleotide exchange found in the 5·8S rRNA‐coding gene. The newly identified genetic variation appears to affect the 5·8S rRNA folding, as deduced from secondary structure models. Additionally, it is shown that for the first time, to the authors’ knowledge, both bursaphelenchid internal transcribed spacers (ITS1 and ITS2) fold into the multibranched closed loops. While the ITS2 closed loop is formed with help of canonical 5·8S‐28S rRNA pairing, the ITS1 forms the thermodynamically stable closed loop with no support of flanking rRNA sequences. The current information on bursaphelenchid ITS rDNA sequence diversity and structure is further discussed. 相似文献
8.
Lin Huang Jian-ren Ye Xiao-qin Wu Xu-ling Xu Jiang-mei Sheng Qing-xia Zhou 《European journal of plant pathology / European Foundation for Plant Pathology》2010,127(1):89-98
A species-specific real-time PCR assay targeting the DNA topoisomerase I gene has been developed to detect Bursaphelenchus xylophilus. The specificity of the assay was confirmed by the lack of amplification of genomic DNA from other Bursaphelenchus or Seinura species. The sensitivity test showed that the limit of the reaction was 0.01 ng of genomic DNA or one individual nematode,
as small as an egg. The validity of the real-time PCR assay was evaluated by analyzing mixed nematode samples extracted from
diseased pine trees in which B. xylophilus was associated with several closely related species, B. mucronatus, B. hofmanni, Aphelenchoides macronucleatus, S. lii and S. wuae. These results demonstrate the potential of the assay to provide rapid, specific and sensitive molecular identification of
B. xylophilus for use in pest risk assessment and quarantine regulations. 相似文献
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11.
J. RAUTAP 《EPPO Bulletin》1986,16(3):453-456
In 1984 a heavy Bursaphelenchus xylophilus infestation was observed in pinewood chips imported from USA and Canada into Finland. Because there are good reasons to suppose that B. xylophilus, if spread to pine forests in North Europe, can cause significant damage, the import of coniferous wood into Finland has been prohibited since autumn 1984. In surveys B. xylophilus has not been found in roundwood or in chips imported from European countries into Finland. Neither has B. xylophilus been detected in Finnish pine trees around harbours nor in paper mills handling North American wood. 相似文献
12.
Bursaphelenchus xylophilus is the causative agent of pine wilt disease. These nematodes disperse in the cortical and wood resin canals of host tree branches in the early stage of infection. To determine whether the dispersal can be regarded as diffusion, we inoculated B. xylophilus into living or boiled Pinus densiflora branch sections and investigated the spatial distribution 24 hr later. Assuming that some nematodes disperse randomly in the axial direction of branch sections and the rest remain at the inoculation site, the coefficient of diffusion (D) by dispersers was estimated to be 23.7–40.7 and 109.4 cm2/day for the living and boiled branches, respectively. The proportion of nondispersers was estimated to be 0.46–0.79 and 0.52 for the living and boiled branches, respectively. When the nematodes were inoculated at the centre of upright living branch sections, they dispersed up and down at similar D values of 7.4 and 7.9 cm2/day, respectively. 相似文献
13.
The pine wood nematode, Bursaphelenchus xylophilus, infects pine trees, leading to fatal pine wilt disease. Here, recombinant venom allergen-like protein (VAP) was obtained by expressing Bx-vap-1 in insect cells. Three-year-old Pinus massoniana were inoculated with recombinant VAP, simulating B. xylophilus esophageal gland secretions. Recombinant VAP up-regulated α-pinene synthase gene expression, the trees showed disease symptoms 15 d after inoculation and the xylem pith revealed brown tissue discoloration, indicating that recombinant VAP could damage P. massoniana cells. Recombinant VAP did not, however, lead to cavitation, indicating that the VAP secreted from B. xylophilus acts as a defense response elicitor. 相似文献
14.
Survival and development of Bursaphelenchus xylophilus (pine wood nematode) was studied for up to 40 weeks in Pinus pinaster sawn wood and branches: 30 boards (1200 × 100 × 25 mm), 30 long‐blocks (1200 × 95 × 95 mm), 10 pine branches with bark and nine branches without bark (1200 mm long). The nematode was found in all materials and through the entire sampling period, with higher abundance in the sawn wood (boards and long‐blocks). In the initial period B. xylophilus reproduced abundantly and a population peak was detected at 8–12 weeks. Subsequently, the populations declined and became dominated by third‐stage resistant larvae (JIII), and in the final sample nematode abundance was very low. Nematode decline in the wood was correlated with a decrease in the moisture content (MC) to below fibre saturation. Survival of the insect vector Monochamus galloprovincialis was also assessed in sawn boards (1200 × 100 × 25 mm, n = 31) and blocks (160 × 95 × 95 mm, n = 40). The majority of the larvae were killed when sawing the wood, although some adults successfully emerged from the boards (10% survival) and blocks (37%). These results represent a contribution to the quantification of the risks of dispersing pine wilt disease through wood packaging materials, confirming that untreated wood can support healthy and abundant B. xylophilus populations for sufficient time for vectors surviving the sawing process to complete their development, to emerge and disperse the nematode. 相似文献
15.
Differences in constitutive and inducible defences in pine species determining susceptibility to pinewood nematode 下载免费PDF全文
下载免费PDF全文 C. S. Pimentel E. V. Gonçalves P. N. Firmino T. Calvão L. Fonseca I. Abrantes O. Correia C. Máguas 《Plant pathology》2017,66(1):131-139
The pinewood nematode, Bursaphelenchus xylophilus, originating from North America (NA), is a major invasive pine pest in Eurasia. It was first detected in Portugal in 1999 associated with maritime pine, Pinus pinaster, and has been differently affecting the main local pine species, P. pinaster and P. pinea. Field studies and direct inoculation experiments in Pinus spp. seedlings, under controlled conditions, were performed to assess whether the differences in constitutive and inducible defences are determining the different susceptibility of pine host species to B. xylophilus. Host co‐evolution with the pathogen was also assessed, including the NA P. radiata, widely used in forestry in the northeast of the Iberian peninsula. Pine mortality in the field was positively related with the abundance of B. xylophilus, and concentration of phenolics and condensed tannins in pines. In the greenhouse assay, seedling tissues were analysed for constitutive investment in defences, as well as the potential inducibility of those defences as driven by B. xylophilus inoculation. Slower growing P. pinea presented higher levels of constitutive defences than faster growing P. pinaster, with only P. pinaster being affected by B. xylophilus. Furthermore, co‐evolution with the pathogen is important, with the fast‐growing NA P. radiata presenting an inducible and effective response to B. xylophilus. Results point to the importance of integrating data on pine life history traits, including growth rate, and production of constitutive and inducible defences, into predictive models for this invasive forest pest. 相似文献
16.
C. S. Pimentel P. N. Firmino T. Calvão M. P. Ayres I. Miranda H. Pereira 《Plant pathology》2017,66(5):856-864
The pinewood nematode, Bursaphelenchus xylophilus, native to North America (NA), is now a major invasive pine pest in Eurasia. Pine species differ greatly in their susceptibility to B. xylophilus, and differences in susceptibility might be related to differences in phytochemistry. This study addressed two major questions: (i) which plant chemical profiles (including compounds that act as plant defences and those important for nutrition) favour pinewood nematode development; and (ii) are NA and Portuguese pinewood nematodes different in their capacity to subsist on plant tissues? For this, B. xylophilus isolates from NA and Portugal were tested on pine phloem and xylem of different pine species. It was found that unaltered pine phloem can sustain the population growth of B. xylophilus, but only in a few cases, and this was clearly related to distinct chemical profiles in some pine species. No evidence was found of evolutionary divergence between B. xylophilus populations in Portugal and NA. The pine species with phloem on which pinewood nematodes could grow tended to have low levels of total phenols, condensed tannins, flavonoids and lignin. Evidence was also found that nutrients such as N may be important. 相似文献
17.
The risks of Bursaphelenchus xylophilus (pine wood nematode) transfer in relation to wood material were assessed. Combinations of infested and non‐infested adjacent boards, long‐blocks and blocks of Pinus pinaster, simulating assembled pallets, were assessed. For the recipient wood, pieces with natural moisture content (MC), heat‐treatment (56°C for 30 min in the core) and kiln‐drying to <20% MC were tested, along with in‐service boards from pallets. Donor and recipient wood materials were kept in direct contact at 25°C or 10°C, with nine replicates per treatment. Bursaphelenchus xylophilus was found to transfer rapidly at 25°C when the wood had an MC above fibre‐saturation point (>30%). Nematode reproduction was rapid and sustained, gradually declining to zero at 40 weeks. Bursaphelenchus xylophilus did not transfer to kiln‐dried or to in‐service wood with an MC below fibre‐saturation point, or to wood at 10°C. The key factors determining nematode transfer were the ambient temperature, the nematode load of the donor wood and the MC of the recipient wood, with a ‘barrier’ of 20% MC below which it becomes unsuitable for nematode transfer. This finding indicates that there is a limited risk of spread of B. xylophilus in treated and untreated solid wood packaging materials. 相似文献
18.
Joana M. S. Cardoso Luís Fonseca Isabel Abrantes 《European journal of plant pathology / European Foundation for Plant Pathology》2012,133(2):419-425
The pinewood nematode (PWN), Bursaphelenchus xylophilus, is the causal agent of pine wilt disease. The international economic impact of the introduction of the PWN into new areas
has highlighted the need for the development of accurate and reliable detection methods of B. xylophilus, which are essential to define aspects of its control and management. In the present study, a methodology was developed for
the direct detection of PWN by conventional PCR assay, with a species specific set of primers based on PWN satellite DNA,
using total DNA extracted directly from maritime pine, Pinus pinaster, wood and bark samples, and from the insect vector, Monochamus galloprovincialis. This methodology involves homogenisation of wood, bark and insects using liquid nitrogen, DNA extraction and one or two
PCR amplification steps, which permit the rapid and direct detection of one single nematode present in 100 mg of wood and
bark and in one entire insect without the preliminary steps of nematode extraction. 相似文献
19.
Pine wilt disease (PWD), recently introduced into Europe, is caused by the pine wood nematode (PWN) Bursaphelenchus xylophilus and is a devastating illness that affects mainly pine trees. It is known that the PWN is capable of infecting other conifers; however, there is currently no information on which other plant species may be susceptible to PWD. In this study, the potential susceptibility of two common species of European forests, Picea abies and Cupressus lusitanica, to PWN was assessed through the monitoring of visual external symptoms, dimension and localization of the nematode population in stems, quantification of total chlorophyll, total soluble phenolics and lignin, at 7, 14, 21 and 28 days after inoculation. The degree of susceptibility was established through the comparison of symptoms with Pinus pinaster, a well‐known PWN host. Furthermore, the stem ultrastructure of P. abies, C. lusitanica and Pn. pinaster was analysed by scanning electron microscopy. The results suggest that P. abies and C. lusitanica are resistant to PWN, and that lignin biosynthesis in these species is affected at an early stage of the infestation. Nevertheless, P. abies seems to be a compatible host that could act as a repository for PWN. 相似文献
20.
C. MAGNUSSON 《EPPO Bulletin》1986,16(3):465-471
Serious concern about the pinewood nematode, Bursaphelenchus xylophilus, and the pine wilt disease arose in 1984 when the nematode was discovered in wood chips imported for the Finnish pulp industry. It has been questioned whether this nematode could establish itself and actually damage pine trees under Nordic conditions. It seems likely that B. xylophilus could reproduce in Nordic forest lands. It is also likely that potential vector insects are present and could be functional in the establishment and epidemic development of the disease. Although temperatures in Nordic areas hardly seem conducive to pine wilt disease, other factors may contribute to disease expression. Research is needed on interactions of stress factors, particularly temperature and soil moisture, in the expression of pine wilt disease. A better understanding of these interactions would aid in evaluating the risk connected with an accidental introduction of the nematode into Nordic countries and provide a sound basis for trade regulations. At present, there are reasons to be most restrictive concerning any possible means of introduction of B. xylophilus. 相似文献