松材线虫病流行与松毛虫危害关系的研究

研究表明 :在松材线虫病发生区 ,其病害的扩展、流行与马尾松毛虫危害有着密切的关系。当年受马尾松毛虫危害的松林 ,枯死松树数量平均为 77 879株 /hm2 ,比未受松毛虫危害的松林 (对照 ) 17 2 73株 /hm2 ,多 35 1% ;次年 ,前者平均枯死松树达 2 0 4 8株 /hm2 ,为对照 2 89株 /hm2 的 6 0 8%。松毛虫危害加速松材线虫病流行的原因是 ,松毛虫危害松树 ,削弱树势 ;弱势的松树招引松墨天牛侵入并增殖加快松树死亡 ;次年大量天牛从“松毛虫危害松木”上羽化 ,侵染健康松林 ,导致松材线虫病进一步扩散和流行。由此认为松毛虫危害是促使松材线虫病流行的重要因子 ,防治松毛虫是控制松材线虫病流行的重要环节之一     

Localized within‐ and between‐tree variation in nematode distribution during latent state of pine wilt disease makes the disease status cryptic

The pinewood nematode (PWN), Bursaphelenchus xylophilus, is the causal agent of pine wilt disease, an epidemic disease that has severely damaged pine forests in East Asia. The disease has spread to northern areas in Asia and parts of Europe. To prevent disease spread as the forefront of damage prevention, a better understanding of infection status is highly important. Not all infected trees show disease symptoms, and such asymptomatic PWN‐carrying trees are likely to be overlooked and can become a pathogen reservoir. To elucidate PWN infection status in asymptomatic trees, we performed PWN inspection of branches and trunks in 21 test trees in two different conditions: trees that had experienced PWN inoculation and those with suspected PWN infection that had experienced transient foliage discoloration. We detected PWNs in eight test trees (38%) and in 13 (1.5%) of a total of 843 samples. The difference in these percentages suggests that nematode inhabitation was highly localized within the trees, possibly owing to the restricted migration of PWNs. Our data demonstrated that trees that were once weakened but recovered their vigour can persist, as the asymptomatic carriers, in the forest. The implications for disease control are also discussed.     

Pine wilt disease: a short review of worldwide research

This article summarizes the results of the research papers presented at the International Symposium on pine wilt disease (IUFRO Working Party Meeting 4.04.03) held in July 2009, at Nanjing, China. The general topics covered were on pine wilt disease (PWD), its causal organism, the pinewood nematode (PWN) Bursaphelenchus xylophilus, plus other PWN-associated microorganisms that play a significant role in PWD such as bacteria (e.g. Pseudomonas fluorescens). Most of the papers that are reviewed are based on work on PWD-PWN in East Asia and Russia. Specific topics covered include: 1) the fundamental conceptions of PWD development, 2) pathogenicity, 3) host-parasite relationships including the histopathology of diseased conifers and the role of toxins from bacteria-nematode ecto-symbionts, 4) PWN life cycle and transmission, 5) B. xylophilus dissemination models, 6) associations (with other nematodes), 7) diagnostics, 8) quarantine and control of the PWN and 9) biocontrol of the PWN.     

A new on‐site detection method for Bursaphelenchus xylophilus in infected pine trees

The pinewood nematode (PWN), Bursaphelenchus xylophilus, is the causal agent of pine wilt disease (PWD), which is a major problem in East Asia and West Europe. Quick identification of PWN is needed to prevent the dispersal of PWD to healthy forests. Various detection methods of PWN have been developed using anatomical characters and molecular markers. These methods are not suitable for rapid diagnosis because it is difficult to distinguish B. xylophilus from the non‐pathogenic species Bursaphelenchus mucronatus based on morphological characters without expertise in nematode taxonomy and most PCR or isothermal amplification detection methods require time‐consuming processes. In this study, we developed an on‐site PWN detection method using a recombinase polymerase amplification (RPA) assay with a novel extraction buffer (DAP buffer). This new PWN detection method is able to extract genomic DNA from PWN in pinewood by simple buffer consisting of sodium hydrate, polyethylene glycol 200 and dimethyl sulfoxide in 10 min without using the experimental devices and able to distinguish between B. xylophilus and other Bursaphelenchus spp. by amplifying the species‐specific 5S rDNA fragment of B. xylophilus in 10 min. Taken together, our protocol can obtain the result for the detection of PWN in pine tree samples within 30 min. This result suggests that RPA/DAP assay is much faster, easier and cheaper than the conventional methods for detecting PWN.     

松林火灾对松材线虫病流行关系的研究

松林火灾与松材线虫的发生和传播有着密切的关系。从火烧松林内羽化的松墨天牛,平均每头可携带７３６４条松材线虫,侵入松林后就会导致松材线虫病的发生和蔓延。火烧松林诱发松材线虫病可分为当年发病型,次年发病型和隐匿发病型。治理火烧松林能有效地控制松材线虫病的发生与传播。根据研究结果,提出了４条新的防治措施和建议。     

Insect vectors of the pinewood nematode: a review of the biology and ecology of Monochamus species

Pine wilt disease (PWD), caused by the pinewood nematode (PWN), Bursaphelenchus xylophilus (Steiner and Buhrer) Nickle 1970 , is a serious threat to susceptible pine forests of the world. The PWN is primarily vectored by Monochamus species (Coleoptera: Cerambycidae). The first occurrence of PWD was reported from Japan in the early 1900s. Following this report, Japanese scientists documented the community of bark‐ and phloem‐inhabiting insects associated with the nematodes in dying trees to determine possible vectors of the nematode. Monochamus alternatus was reported to be the most effective vector in Japan. The primary vector in North America is Monochamus carolinensis, and in Europe, it is Monochamus galloprovincialis. Further studies have been expanded through the nematode‐invaded countries of Korea, Taiwan, China and Portugal. There is an interspecific association between the PWN and its insect vectors, and it is an obligatory component of the disease cycle. It is crucial to understand this relationship as well as the population ecology of the beetle to aid in monitoring and control of this worldwide threat to pine forests. Studies to date indicate a remarkable similarity among beetle species around the globe for a variety of life‐history traits, including lifespan, adult emergence numbers, flight capability, nematode transmission rates and attraction to pine volatiles. Wherever pines are found, there is a beetle species capable of transmitting the nematode. Although flight performance and range is generally poor for this group of beetle vectors, the cryptic nature of the species and the lack of interest in the beetles by countries in the absence of the nematode have led to the disease establishing a foothold in a variety of countries such as Portugal. In this paper, studies conducted in different countries on Monochamus vector species of the PWN are compared and discussed.     

Pine wilt disease: detection of the pinewood nematode (Bursaphelenchus xylophilus) as a tool for a pine breeding programme

The pinewood nematode (PWN), Bursaphelenchus xylophilus, is a serious quarantine pest first detected in Portugal and Europe in 1999. It is the causal agent of pine wilt disease (PWD). A resistance breeding programme has been initiated to contribute to control the evolution of the disease. Five hundred and four adult maritime pine, Pinus pinaster, trees were phenotypically selected as candidate trees for this programme from an area affected by PWD. To identify tolerance to the nematode, the selected trees were monitored monthly. Over the course of 1 year, 57 candidate trees died and were tested for the presence/absence of the PWN. As accuracy of detection is of major importance, an ITS‐PCR‐based method applied directly to wood from adult maritime pine trees was tested and compared with a standard morphological identification method. The results showed that the use of PCR to detect the pathogen provided more rapid and accurate results in comparison with the standard morphological identification. Thus, this method is suitable to be used in the survey of the breeding population for resistance/tolerance to PWD.     

Suggestions on management measures of pine forest ecosystems invaded by <Emphasis Type="Italic">Bursaphelenchus xylophilus</Emphasis>

Pine wood nematode （PWN）, Bursaphelenchus xylophilus （Steiner et Buhrer） Nickle is an important invasive alien species in forests of China and has become one of the most destructive forest diseases. In order to improve the resistance and resilience of pine forest ecosystems against B. xylophilus invasion and make the pine forest ecosystem more timely responsive to PWN invasion, we made some recommendations based on five years of intensive observations. We advocate a set of management measures with the theme “Prevention is priority, but integrated with curative techniques and ecological resilience” on the pine forest ecosystem invaded by B. xylophilus; details of accurate measures are proposed. The aim is to discover the underlying problems of present pine forest ecosys-tems and to take, correspondingly, administrative measures and strategies, which will encourage the pine forest ecosystem to develop in a benign way.     

松材线虫病发生34 a的综合防控——以江苏省镇江市为例

镇江市于1985年发现松材线虫致病松树以来,该病迅速扩散蔓延,给当地的经济和生态造成了重大损失,成为江苏省松材线虫病发生最为严重的地区之一,镇江市多个县、区为松材线虫病疫区。自从发生松材线虫病疫情以来,镇江市积极采取行动,采取了一系列综合防控松材线虫的方法。包括开展松材线虫病普查工作、加强松材线虫检疫、感病松林病死松木清理、传播媒介林间施药防治和引诱防治、生物防治以及开展营林措施防治。经过多年的防控实践,从2009年开始,镇江市各辖市、区松材线虫病的发生面积和病死树数量都呈现双下降趋势,镇江市松材线虫防治工作取得了明显的成效。     

Modelling of the spread of Dothistroma septosporum in Europe

Dothistroma needle blight (DNB), a disease affecting several pine species, is currently generating great concern in Europe. Caused by Dothistroma pini and Dothistroma septosporum, DNB affects pine needles and causes premature defoliation, which results in growth reduction and, in extreme cases, mortality. The disease has increased in importance in Europe over the last 20 years, with an increase in the number of observations of DNB in regions with large areas of Pinus sylvestris in northern Europe. This article presents a cell‐based spatiotemporal model for predicting the likelihood and intensity of the future spread of D. septosporum in Europe. Here, “spread” includes both invasion of new regions and infection of healthy stands within already‐colonized regions. Predicted spread depends on the availability of host species, climatic suitability of different regions to D. septosporum and dispersal of sexual and asexual spores from infected trees to surrounding forests via water splash, mist and wind. Long‐distance spread through transport of infected seedlings is also included in the model. Simulations of spread until 2007 and 2015 were used to validate the model. These simulations produced similar patterns of spread to those observed in Europe. Simulations for 2030 suggested that additional and new outbreaks are likely to occur in Scotland, southern Norway, southern and central Sweden, northern parts of Germany and Poland, Estonia, Latvia and south‐west Finland. Preventing the delivery of infected seedlings would be an effective method for reducing the spread of D. septosporum in the Nordic countries, Scotland and Ireland, the Baltic countries, and parts of Germany, Poland and Belarus. In these states, prevention of transport of infected seedlings can reduce the probability of additional spread by 15%–40%.     

松兰山景区松材线虫病综合治理研究

研究分析了浙江宁波市象山松兰山景区松材线虫病发生规律与防治效果。结果表明：只采取“封锁”措施，会使松材线虫病迅速爆发毁灭全林。从1992年到1998年，松兰山枯死松树从6株增到5893l株，增长了9820．8倍；采用皆伐松林防治松材线虫病并改造成板栗、杨梅的方法，会导致逆向演替，使松林最后成为以疏花野青茅为主的荒山。而采取综合防治措施就可有效地控制松材线虫病的危害。从1998年到2002年，松兰山枯死松树下降到137株，缩小了430倍，使松兰山仍然保持着葱郁的松林群落。     

开展松材线虫病抗性选育 保护我国松林资源

松材线虫病是我国松林毁灭性病害,已造成数千万松株死亡,发生区经济损失巨大,目前此病害正威胁着南方重点林区及重要风景名胜区松林的安全。我国松属资源丰富,科技人员队伍组织体系完备,抗性选育研究已有较好基础。因此尽快开展抗性选育有利于发生区森林恢复、非发生区林分改造和结构调整,有利于保护我国森林资源。本文对开展抗性选育提出了具体建议。     

松材线虫病流行与松树采伐剩余物关系研究

研究表明，在松材线虫发生区，疫病的扩散和流行与松树的采伐剩余物有着密切的关系。松树的新鲜采伐剩余物，如梢，枝，干，桩，每平方分米表面积松褐天牛幼虫蛀入孔数分别为1．46个，1．41个，0．18个，0．29个。次年，从这些剩余物上每平方分米分别能出现0．55个，0．84个，0．15个，0．27个松褐天牛羽化孔，经镜检，受松褐天牛寄生的采伐剩余物均带有松材线虫。由此可见，松树采伐剩余物是松材线虫病扩散流行的一个重要的中间环节，改善松林卫生状况，规范松林采伐技术，科学处理剩余物是抑制松材线虫病扩散和流行的重要环节和措施。     

Comparison of histological responses and tissue damage expansion between resistant and susceptible Pinus thunbergii infected with pine wood nematode Bursaphelenchus xylophilus

Pine wilt disease caused by the pine wood nematode (PWN), Bursaphelenchus xylophilus, has been epidemic and has had disastrous impacts on pine forests and forest ecosystems in eastern Asia. Many pine species in this area are susceptible to this disease. Pinus thunbergii is particularly susceptible. In Japan, tree breeders have selected surviving trees from severely damaged forests as resistant candidates, and have finally established several resistant varieties of P. thunbergii. However, this breeding procedure requires much time and effort due to the lack of physiological and phenotypical information about resistance. To investigate the resistance mechanisms of selected P. thunbergii, we compared histochemical responses, tissue damage expansion, and PWN distribution in resistant and susceptible clones of P. thunbergii after PWN inoculation. The results suggested that the mechanisms of resistance are as follows: damage expansion in the cortex, cambium, and xylem axial resin canals are retarded in resistant trees soon after inoculation, probably due to the induction of wall protein-based defenses. Suppression of PWN reproduction was particularly caused by inhibition of damage expansion in the cambium. The slow expansion of damage in each tissue provides time for the host to complete the biosynthesis of lignin in the walls of cells that surround the damaged regions. This lignification of cell walls is assumed to effectively inhibit the migration and reproduction of the PWNs. The mechanism of initial damage retardation is presumed to be a key for resistance.     

Direct PCR‐based method for detecting Bursaphelenchus xylophilus,the pine wood nematode in wood tissue of Pinus massoniana

Pine wilt disease (PWD), caused by the pine wood nematode (PWN) Bursaphelenchus xylophilus, leads to serious losses to pine forestry around the world. Pinus massoniana, which is vulnerable to be attacked by the PWN, is the dominant species used in pine forestry in China. The objective of this study is to develop a direct PCR‐based method for detecting B. xylophilus in the wood of P. massoniana without a separate nematode extraction step. A simple procedure was first developed for isolating B. xylophilus DNA in 5 mg pine wood tissue samples harbouring PWN for detection by PCR amplification. A B. xylophilus‐specific amplicon of 403 bp (DQ855275) was generated by PCR from the infested wood tissue. The entire procedure can be completed within 5 h with one pair of primers. This assay can serve as a rapid, cheap and environmentally friendly method to detect B. xylophilus in samples of P. massoniana.     

Management of pine wilt disease in Korea through preventative silvicultural control

Pine wilt disease (PWD) represents a major threat to forest ecosystems worldwide. Although PWD is now better understood, effective control measures for this disease have still not been devised. Here, we report several years of field studies on preventative silvicultural control of PWD. Silvicultural control through preventative clear-cutting and the manual removal of logs was implemented between 2005 and 2009 in 16 Korean districts that had newly PWD-infected stands. Preventative clear-cutting of neighboring asymptomatic pine trees (within a 10-50-m radius of wilt trees) and the removal of felled logs or branches suppressed spread of PWD. Occurrences of PWD wilt pines in districts (city or county) subjected to this silvicultural control method were significantly reduced compared with those in districts using conventional controls (physical or chemical treatment of wilt pine trees). Through silvicultural control, PWD was successfully suppressed in 11 of 16 districts investigated. In contrast, successful control was achieved in only 1 among 18 districts subjected to conventional control. Our results will be of considerable interest to those engaged in the very difficult battle against the global spread of PWD.     

Within-tree distribution and attractant sampling of propagative pinewood nematode, Bursaphelenchus xylophilus: An early diagnosis approach

Early detection is of primary importance to enable rapid actions to prevent the spread and introduction of invasive species. The pinewood nematode (PWN), Bursaphelenchus xylophilus, a serious invasive and destructive species, is listed as a quarantine pest in the legislation of more than 40 countries. However, Baermann funnel extractions of wood from discs cut from trees at breast-height often do not detect the presence of PWN in infested trees. A serious consequence of such false negatives is the loss of the best window for implementation of eradication or quarantine measures to prevent establishment of incipient PWN infestations. Here we document the within-tree horizontal and vertical distribution of PWN in infested stands in China, using a newly developed kairomonal trapping technique. Our results provide a simple, effective, rapid and non-destructive sampling method that takes into account the changes of PWN within-tree distribution in relation to pine wilt disease (PWD) symptom development. When 60–80% of the foliage has become pale green, PWN is recovered from larger diameter branches. As disease symptoms progress, PWN moves into and down the trunk. As the needles turn yellow, PWN was recovered from the trunk at 1–2 m above the ground. The correlation between the within-tree distribution of PWN and the expression of symptoms indicated a strong association between the distribution of PWN and physiological and pathological changes that develop in attacked pines through the interaction between PWN and tree. This systematic sampling technique takes into account the within-tree distribution of the nematode and should greatly enhance early detection of PWN in field surveys, monitoring and phytosanitary inspections.     

韩国松材线虫病防控成效及启示

由松材线虫引起的松材线虫病对松林构成了严重威胁,松材线虫病的防控受到社会极大关注。30多年来,韩国为了遏制松材线虫病的扩展蔓延,减少灾害损失,保障生态安全,对松材线虫病采取立法及检疫、化学防治、清理病死树及疫木无害处理、预防性造林、检测和监测等措施,取得了良好的防治效果。文中回顾韩国松材线虫病发生过程,分析其流行特点,介绍防控措施及成效和近期研究现状,提出韩国松材线虫病防控成效对我国松材线虫病研究与防控的主要启示,即制定和完善松材线虫病防治相关法规,分类施策、综合治理,建立适合我国国情的松材线虫病防控体系和加强科学研究,提高科技支撑水平。     

Pinus pinaster Ait. tree mortality following wildfire in Spain

Maritime pine (Pinus pinaster Ait.) is the tree species most affected by wildfire in the Iberian Peninsula. Prediction of the probability of fire-injured tree mortality is critical for management of burned areas, evaluation of the ecological and economic impact of wildfire and prescribed fire planning and application. Pine bark beetles (Scolytidae) frequently attack burned maritime pine stands and cause extensive post-fire mortality throughout the Iberian Peninsula. In the present study, maritime pine trees were monitored for three years following 14 wildfires in four ecotypes in Spain (11 fires in Galicia (Galician ecotype - NW Spain), one fire in Portillo (Meseta-Castellana ecotype - Central Spain), one fire in Rodenal (Rodenal ecotype - Central Spain), and one fire in Genalguacil (Sierra Bermeja ecotype - SW Spain)). Data on tree attributes, crown and bole injury, ground fire severity, Ips sp. presence and tree survival were obtained by examining 3085 trees. Logistic regression models for predicting the probability of delayed maritime pine mortality were developed by use of generalized estimated equations (GEE). An ample range of response to fire damage in mortality was evident among the four ecotypes and different models were fitted for each. The most important variables for predicting tree mortality were total crown volume damaged, presence of Ips sp. attack and cambium kill rating. The results highlight the extensive presence of Ips sp. in burned maritime pine forests and its importance in tree mortality process, the ample range of response of P. pinaster, in terms of post-fire mortality, as well as the need to develop site specific mortality models for the different ecotypes of this species following fire.