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1.
Since the beginning of the 20th century, pine trees in Japan have been seriously damaged by the pine wilt disease. This disease is caused by the pine wood nematode, Bursaphelenchus xylophilus, which is transmitted by the Japanese pine sawyer, Monochamus alternatus. The control of disease depends to a large extent on chemicals, but the public is now demanding environmentally friendly control methods. The virulence of B. xylophilus varies very widely. Pre-inoculation of young pine trees in a nursery with avirulent B. xylophilus has induced systemic resistance of trees against a subsequent inoculation with virulent B. xylophilus. This induced resistance was considered a hopeful means for developing a biological control for the disease. The induced resistance by the avirulent nematodes was also expressed in mature pine trees in a forest where the disease was naturally epidemic. However, the effects of induced resistance were not satisfactory for practical biological control. Since the inoculation with higher concentrations of the avirulent B. xylophilus induced the resistance more effectively, the pre-inoculation method will need to be improved to develop the biological control. The induced resistance of pine trees by avirulent B. xylophilus should be one of the candidate biological control methods against pine wilt disease. This induced resistance also provides an experimental system to clarify physiological interactions between the nematodes and pine trees.  相似文献   

2.
为筛选出新型的高效松材线虫Bursaphelenchus xylophilus防治药剂,采用浸渍法室内测定了氯氟氰虫酰胺、阿维菌素、甲维盐、灭多威、特丁硫磷、甲基异柳磷和吡虫啉7种药剂对松材线虫的毒杀作用和对其繁殖能力的影响,采用高效液相色谱仪测定氯氟氰虫酰胺在马尾松体内的输导分布,并在林间开展树干注射氯氟氰虫酰胺抗松材线虫病试验。结果表明,7种药剂中氯氟氰虫酰胺对松材线虫的毒力最强,LC_(50)为0.019 mg/L,阿维菌素和甲维盐次之,LC_(50)分别为0.0213 mg/L和0.0214 mg/L。氯氟氰虫酰胺树干注射3个月内能迅速从马尾松基部传导至树梢顶部,树干边材东、南面的药剂含量显著高于西、北面,且树干不同高度的氯氟氰虫酰胺含量均高于其LC50;氯氟氰虫酰胺制剂在林间注干后,对人工接种病原松材线虫松树的防治效果为73.33%,自然感染松材线虫病松树的死亡率降至2.00%,而对照试验中人工接种松材线虫松树和自然感染松材线虫病松树的死亡率分别为93.33%和19.67%。表明氯氟氰虫酰胺制剂能有效预防松树个体的松材线虫病发生与为害,可以推广使用。  相似文献   

3.
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   

4.
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   

5.
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   

6.
树栖真菌对扩散型松材线虫生活史的影响   总被引：1，自引：0，他引：1  
 松材线虫病是一种涉及多个生物因子的复杂病害系统,其病原松材线虫的生活史受媒介昆虫、寄主、树栖真菌和伴生细菌等多种因素影响。利用感染松材线虫病的马尾松林内的两种优势内栖真菌的3个菌株,松球壳孢(Sphaeropsis sapinea)E11、MHS7.3和小孢拟盘多毛孢(Pestalotiopsis microspora)M32,接种于松褐天牛模拟蛹室,研究松褐天牛蛹期和羽化过程松材线虫种群动态,以及松褐天牛携带扩散型松材线虫的动态变化。结果表明,天牛发育处于蛹期时,松材线虫主要以扩散型三龄幼虫(JⅢ)附着于天牛体表;发育至羽化期时,松材线虫主要以扩散型四龄幼虫(JⅣ)位于天牛体内。不同优势内栖真菌接种处理下,松褐天牛携带的扩散型幼虫数量的大小顺序为:小孢拟盘多毛孢M32>松球壳孢E11>松球壳孢MHS7.3,这与模拟蛹室中松材线虫种群的大小顺序是一致的,表明松褐天牛携带的线虫数目与蛹室中线虫种群的大小关系密切。在松褐天牛携带的JⅢ、JⅣ和扩散型幼虫(JⅢ＋JⅣ)占线虫种群比例方面,松球壳孢E11和MHS7.3均大于小孢拟盘多毛孢M32,显示不适宜繁殖的真菌能促进群体内更多个体转化为扩散型虫态。JⅣ在蛹期的第二天即从天牛体内分离得到,但数量极少,随着JⅢ在羽化前达到最大值,JⅣ也开始大量出现,预示着较大JⅢ种群数量可能是JⅣ大量发生的基础条件之一。研究结果表明在松材线虫的扩散过程中,松林优势树栖真菌对媒介天牛携带的松材线虫数量产生显著影响。  相似文献   

7.
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   

8.
BACKGROUND: Pine wilt disease (PWD) is very complex and has been reported to be caused by pine wood nematode, Bursaphelenchus xylophilus (Steiner & Buhrer) Nickle, and its accompanying bacteria. However, there is no report on the control of PWD by antibacterial agent. The present study was performed to investigate disease control efficacy of antibacterial agents against PWD. RESULTS: Among six antibacterial antibiotics tested, oxolinic acid (OA) showed the strongest antibacterial activity against five bacteria isolated from three strains of pine wood nematode. In in vivo assay, it effectively suppressed the development of PWD in three‐year‐old seedlings of Pinus densiflora Sieb. & Zucc.; it showed 71% control when injected at 3 mg per seedling. A mixture of OA and the nematicidal agent abamectin (Ab) showed higher disease control efficacy against PWD than either OA or Ab alone. In addition, OA alone and a mixture of OA and Ab also controlled PWD in approximately 20‐year‐old pine trees under field conditions. CONCLUSION: This is the first report on the suppression of PWD by OA. The result strongly indicates that PWD could be controlled by antibacterial antibiotic alone and a combination of antibacterial and nematicidal agents. Copyright © 2010 Society of Chemical Industry  相似文献   

9.
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10.
Bursaphelenchus xylophilus, the pinewood nematode and causal agent of pine wilt disease (PWD), is a globally destructive pine pest. PWD is favoured by high temperatures and drought, but B. xylophilus can also persist in symptomless hosts in areas not prone to PWD. We compared seedlings of different Pinus species that were inoculated with B. xylophilus when well-watered and under moderate temperatures, conditions that favour sublethal infections. We compared isolates of B. xylophilus in south-west Europe (Portugal) and south-east USA (Louisiana). P. pinea, P. pinaster, and P. radiata were challenged in both areas with inoculations of local B. xylophilus. P. sylvestris in Portugal and P. palustris in Louisiana were also tested. Seedling mortality was low, as expected, but nematode establishment was common. There were higher nematode densities and more damage in seedlings of P. sylvestris and P. radiata, followed by P. pinaster, with P. pinea and P. palustris proving to be more tolerant. Life history of hosts was a better predictor of resistance to B. xylophilus than geographical origin, even under sublethal conditions. Fast-growing species (P. radiata and P. pinaster) appeared most likely to be symptomless carriers in areas not prone to the development of PWD. Cold-adapted species (P. sylvestris) may still be suitable hosts for the pathogen, even at the sublethal conditions of their distribution areas. There were more nematodes per gram of host using nematode isolates from Portugal. Nonetheless, our results opposed the hypothesis that North American pinewood nematodes have such low virulence that they are obligatorily saprophytic.  相似文献   

11.
Pine wilt disease is currently among the most devastating pine tree plagues on earth. It is caused by the pinewood nematode Bursaphelenchus xylophilus in a perfect and well synchronized mutualistic relationship with beetles of the genus Monochamus. The disease has a fast and efficient mode of spread, which makes most of the efforts to control it practically insufficient. We investigate how early eradication of infected pine trees, i.e. eradication of trees which just ceased oleoresin exudation, may affect disease spread. In contrast to the sole eradication of killed trees, our results show that under an appropriate combination of eradication strategies: (1) there is a significant increase in the minimum pine density below which the disease fails to invade; (2) the region where reproductive Allee effects may take place are significantly enlarged. We implement optimal strategies for eradication through stochastic search optimization techniques, and conclude that disease extinction can be reached faster with an appropriate combination of eradication measures, minimizing the damage on healthy pine trees and operational costs.  相似文献   

12.
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   

13.
Pine wilt disease (PWD) is the most destructive disease threatening pine worldwide. The disease is mainly caused by the pine wood nematode, Bursaphelenchus xylophilus, which is vectored by pine sawyer longhorn-beetles, Monochamus spp. This study aimed to select resistance-inducing pine endophytic bacteria for management of PWD. To set up a defence-related genes expression pattern for screening, four chemical inducers (salicylic acid, γ-aminobutyric acid (GABA), β-aminobutyric acid and α-aminobutyric acid) were tested in vitro on pine calli and in vivo on pine seedlings. Treatment with GABA had the greatest reduction in PWD severity on pine seedlings. The pattern of defence-related gene expression in calli treated with GABA was used to select potential resistance-inducing bacterial strains. In addition, 92 bacterial strains were isolated from pine tree needles and stems and were tested for expression of defence-related genes in pine calli in vitro. Among the tested strains, 13 showed a similar pattern to GABA treatment in at least four tested defence-related genes and were selected for the seedling assay. From the seedling assay, three bacterial strains (16YSM-E48, 16YSM-P180 and 16YSM-P39) showed significant reduction in PWD severity compared to the untreated control. Moreover, among the selected strains, cell-free culture supernatant of strain 16YSM-P180 significantly reduced PWD severity in inoculated pine seedlings. The selected strains were identified based on the 16S rRNA sequence as Pseudomonas putida 16YSM-E48, Curtobacterium pusillum 16YSM-P180 and Stenotrophomonas rhizophila 16YSM-P39. These selected strains are suggested as potential alternatives for management of PWD by induction of systemic resistance.  相似文献   

14.
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   

15.
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   

16.
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   

17.
Á. Tóth  M. Elekes 《EPPO Bulletin》2013,43(1):152-154
An ongoing official survey to detect the pine wood nematode Bursaphelenchus xylophilus, a quarantine pest, started in 2003 in coniferous forests in Hungary. Based on the results of the study from 2003–11, B. xylophilus has not yet been detected in Hungary. Two other Bursaphelenchus species (B. mucronatus and B. vallesianus) were identified in samples in 2009. Details of the survey and the measurements of B. mucronatus and B. vallesianus are provided.  相似文献   

18.
Monochamus galloprovincialis Olivier (Coleoptera: Cerambycidae) is one of the vectors of the pinewood nematode Bursaphelenchus xylophilus (Steiner and Buhrer) Nickle (Nematoda: Parasitaphelenchidae), the causal agent of pine wilt in susceptible pine trees. It is therefore important to study both the biological and ecological characteristics of the beetle. In this study reproductive potentials of female beetles reared on two pine species were investigated under laboratory conditions and compared using fertility tables constructed for beetles reared on each tree species. Beetle population parameters were compared between tree species. The intrinsic rate of increase and associated population parameters of beetles reared on Pinus sylvestris L. were consistently higher than those reared on P. nigra Arnold. This result suggests that P. sylvestris has a significantly greater positive effect on the reproductive potential of M. galloprovincialis populations than does P. nigra.  相似文献   

19.
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   

20.
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   

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