Response of pine forest to disturbance of pine wood nematode with interpretative structural model

Pine wood nematode (PWN, Bursaphelenchus xylophilus), originating from North America, causes destructive pine wilt disease. Different pine forest ecosystems have different resistances to B. xylophilus, and after its invasion, the resilience and restoration direction of different ecosystems also varies. In this study, an interpretative structural model was applied for analyzing the response of pine forest ecosystem to PWN disturbance. The result showed that a five-degree multi-stage hierarchical system affected the response of the pine forest ecosystem to PWN disturbance, in which direct affecting factors are resistance and resilience. Furthermore, the analysis to the 2nd, 3rd and 4th degree factors showed that not only does distribution pattern of plant species and pine’s ecological features affect the resistance of pine forests’ ecosystem, but removal of attacked trees and other measures also influence the resistance through indirectly affecting the damage degree of Monochamus alternatus and distribution pattern of plant species. As for resilience, it is influenced directly by soil factors, hydrology, surrounding species provenance and biological characteristics of the second and jointly dominant species, and the climate factors can also have a direct or indirect effect on it by affecting the above factors. Among the fifth elements, the elevation, gradient and slope direction, topographical factors, diversity of geographical location and improvement of prevention technology all influence the response of pine forest ecosystem to PWN disturbance. __________ Translated from Scientia Silvae Sinicae, 2007, 43(8): 85–90 [译自: 林业科学]     

松材线虫入侵在生态系统水平上的研究现状与展望

松材线虫是由外来入侵种松材线虫做病原、天牛做媒介,引起的一种林木病害,对我国的松林生态系统造成了严重的破坏,也给我国的经济造成了巨大的损失。对生物入侵种松材线虫国内外的研究现状进行综述,指出了松材线虫病研究集中的几个方面,进一步指出松材线虫入侵在生态系统水平上的研究,提出了在生态系统水平的研究方向。     

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.     

Differences in arthropod communities between island and inland Masson pine forests infested by pine wilt disease in Zhejiang Province

The invasion of Bursaphelenchus xylophilus （pine wood nematode, PWN） carried by Monochamus alternatus predominately attacks Masson pine （Pinus massoniana） forests and causes great economic losses in China. In this study, we assessed whether the effect of the invasion of PWN is different between island and inland forests. Arthropods were sampled in Fuyang （inland） and Zhoushan （island） counties in Zhejiang Province with sweep netting and light traps at four plots. During two field periods （May to June 2004 and September to October 2005） a total of 21,916 insects, representing 384 species belonging to 99 families and 15 orders, were collected in the sample plots from the island, whereas, from the inland forest a total of 29,262 insects, representing 308 species belonging to 96 families and 13 orders, were found. A hierarchical cluster analysis （HCA） and one-way ANOVA, based on the composition of different arthropod guilds, were performed. The results showed that there was no significant difference in the composition of arthropod communities at the family level between inland and island. But these two habitats had a significant effect on the composition of species, individuals, sub-communities and energy class levels. Statistically, the composition of the two orders, Lepidoptera and Diptera, in the two habitats were significantly different.     

Effect of pine wood nematode invasion on pine community functions in the Pinghu region, Zhejiang Province, eastern China

In order to investigate the effect of invastion by pine wood nematode （PWN）, this study analyzed severalfunctional indices, i.e., the increment in DBH and stand volume and biomass, in the damaqed stands with various mixedpercentages of Pinus massoniana and P. thunbergii and with different levels of damage. According to the results of rate of change in increment of DBH and stand volume, the forest ecosystem resistance against PWN increased with a reduct on n the m xed ratio of pine. The resistance was highest with a mixed percentage of 50%. The invasion of PWN hanged the corresponding relationship of increment between DBH and stand volume （pure stands 〉 7：3 conifer and roadleaf 〉 6：4 conifer and broadleaf 〉 5：5 conifer and broadleaf） among the P. thunbergii stands when there is no amage, but for P. massoniana stands this phenomenon did not occur. For the increment rate of DBH and stand volume, is significant change in P, thunbergii forest indicates that the resistance of pure P. thunbergii forest was higher than at of P. massoniana. The invasion of PWN accelerates the succession from pure stands to mixed stands and then tohe broadleaf evergreen stands.     

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。     

First isolation of pine wood nematode from Pinus tabuliformis forests in China

Previous risk prediction studies have mostly shown that suitable areas for pine wood nematode, Bursaphelenchus xylophilus (PWN), in China are mainly concentrated in eastern and southern China, with pest reaching no further northern than the Yangtze River basin. The Chinese pine (Pinus tabuliformis, Chinese pine or Chinese red pine) is the main, native, afforestation species in areas to the north of the Yangtze River basin. To our knowledge, there is no report on PWN infecting Chinese pines in the north of the Yangtze. In this study, we used Baermann funnel method and several PCR primers to detect the presence of PWN in Chinese pine forest fields in 2010 in Shaanxi, China, which is far to the north of the Yangtze River and a cause for concern. Pinus koraiensis (Korean pine) may also be at risk and should also be monitored to prevent invasion by PWN.     

保松灵防治松墨天牛的林间试验

松材线虫病 [Bursaphelenchus xylophilus (Steiner et Buhrer) Nickle]严重威胁我国的森林资源。浙江等地每年因该病造成大量松树枯死 ,防治松材线虫病刻不容缓。当前 ,我国防治该病主要措施是清理枯死松树 ,降低媒介昆虫松墨天牛 (Monochamus alternatus Hope)密度 ,减缓病害扩散蔓延。但在风景名胜区的松林和古树名木等 ,一旦发生该病 ,仅清理死树是不够的 ,必须采取保护性措施 ,防止因病害导致大量宝贵松树枯死。在日本 ,长期以来主要措施是在松树树冠进行空中或地面喷洒杀螟松、倍硫磷、西维因等农药 ,杀灭松墨天牛成虫 ,从而防治松…     

The population structure and sex ratios of Bursaphelenchus xylophilus under a-pinene stress

Pine wood nematode(PWN), Bursaphelenchus xylophilus, is a serious pathogen of pines throughout the world. Previous work indicated that different concentrations of a-pinene could affect nematode reproduction,however the mechanism of that influence is not clear. In order to examine the reproductive strategies of PWN in response to the stress of the volatile material a-pinene, we investigated different aspects of population changes of B.xylophilus under two concentrations of a-pinene. The results show that a high concentration(214.5 mg ml-1)promoted population growth while a low concentration(56.33 mg ml-1) decreased the population. Population structure was analyzed and it was found that a high concentration of a-pinene decreased the percentage of adults but increased the percentages of larvae and eggs.Furthermore, from the results of an evaluation of sex ratios(female/male), it was determined that a high concentration could elevate sex ratios but a low concentration decreased ratios sharply. These results suggest that the PWN could regulate its population by changing sex ratios under stress of a-pinene. This study has provided a theoretical basis for the prevention and control of pine wilt disease caused by the pine wood nematode.     

松材线虫病的潜伏侵染及松墨天牛传播新途径

在南京用松材线虫接种 7种松树 ,于第 2a在无任何症状的松树的接种点上方 10cm处取样 ,分离线虫。结果表明松材线虫病的潜伏侵染现象比较普遍。在不同的松树品种上潜伏侵染现象有所差异。感病黑松和赤松很快死亡 ,只有在接种量小的情况下才有潜伏侵染现象发生。抗病性强的火炬松和湿地松感病后潜伏侵染现象比较普遍 ,并且样品中的线虫量也较高。而抗病性中等的马尾松和刚松感病后潜伏侵染现象的普遍性及样品中的线虫量均处于中等。短针松接种松材线虫后既不枯死 ,也无潜伏侵染现象。松墨天牛传播松材线虫的试验结果表明 ,未携带松材线虫的松墨天牛成虫在松材线虫病松枝上取食后 ,再到健康松枝上取食 ,有传播松材线虫的可能。这些结果表明在松材线虫病新病区清理病死木时 ,应采取早期诊断技术 ,把未表现症状的松树一起清除 ,以达到防治目的。     

An inoculation experiment of Japanese <Emphasis Type="Italic">Bursaphelenchus</Emphasis> nematodes on Japanese black and red pine, <Emphasis Type="Italic">Pinus thunbergii</Emphasis> and <Emphasis Type="Italic">P. densiflora</Emphasis>

The pinewood nematode (PWN) Bursaphelenchus xylophilus is an invasive pathogen that was introduced from North America to Asian countries and Portugal and is devastating native pine forests. Some native European and Asian Bursaphelenchus nematodes also have weak to moderate pathogenicity to native pine species. To evaluate the potential risk of native Bursaphelenchus species, we inoculated ten Japanese Bursaphelenchus species into native pine species (the dominant forest species) in Japan, and evaluated their pathogenicity using mortality and tracheal tissue damage as indices. Inoculation was conducted on August 3, 2007, and the symptoms were observed every 2 weeks until February 1, 2008. None of the inoculated trees, excluding the pathogenic PWN inoculated control, showed external disease symptoms; however, four species [a less pathogenic PWN isolate, B. luxuriosae, Bursaphelenchus sp. NK215 (undescribed), and NK224 (undescribed)] caused tracheal tissue damage in inoculated seedlings and showed weak pathogenicity. Therefore, we conclude that there are some potentially pathogenic native species of nematodes distributed in Japan. Interestingly, two of these weakly pathogenic species, B. luxuriosae and NK215, are not associated with Pinaceae trees, suggesting that nematode pathogenicity may be a pre-adaptive character. More experimental studies under different conditions are necessary to accurately evaluate the potential risk of these pathogens.     

日本松材线虫病治理技术考察

日本是世界上松材线虫发生最早、受害损失最严重的国家，也是对松材线虫研究最深入、控制经验最丰富、治理技术最先进的国家。通过对该国多个林木育种中心、森林综合研究所及基层森林经营管理机构、生产者等的访问交流和实地考察，较系统地介绍了松材线虫病在日本的发生危害、研究现状和实用控制技术，提出了有效预防、治理松材线虫病的措施和方法。     

A review of the development of Mediterranean pine–oak ecosystems after land abandonment and afforestation: are they novel ecosystems?

Context

Mediterranean landscapes are composed of different interacting vegetation patches. Pine and oak ecosystems form contiguous patches within these landscapes, in pure stands, or as mixed pine?Coak ecosystems. During the nineteenth century, pine forest distribution in the Mediterranean Basin increased dramatically as a result of large-scale re-forestation and spontaneous forest regeneration. At the same time, secondary succession of abandoned agricultural land allowed development of pine and oak ecosystems. Consequently, a pine?Coak mosaic has developed, which created opportunities for cross-colonization, i.e. species colonization from one ecosystem in the reciprocal system. Pines shed their wind-dispersed seeds and colonize Mediterranean oak vegetation. Oaks regenerate in different ecosystems, including pine forest understories.

Research question

This paper reviews fire-free landscape-scale dynamics of pine?Coak Mediterranean mosaics and analyze how landscape-scale interactions are leading to pine?Coak ecosystems by different processes.

Results

Published information from the Mediterranean Basin illustrates pathways of pine?Coak ecosystems formation. Using Mediterranean literature, I try to elucidate the factors that (1) control colonization potential and (2) modulate the resistance to colonization, in different habitats, land uses, and landscape settings.

Conclusion

Management implications for these mixed pine?Coak ecosystems are suggested. The question of whether they are novel ecosystems is discussed.     

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。     

几种诱杀松墨天牛方法的效果比较及评价

利用３种方法诱杀松墨天牛成虫的试验结果表明,松饵木能引诱到补充营养的松墨天牛成虫,且雌成虫能在松饵木上产卵,引诱剂对引诱怀卵的松墨天牛成虫的效果较好。引诱剂和松饵木使用方便、安全,成本较低,大面积应用可降低松墨天牛种群密度,减少松材线虫传播机会。但单独使用达不到防治松材线虫病的目标。灯光诱杀远不如引诱剂和松饵木,且其成本高,安全性差,用于松墨天牛防治不宜提倡。利用引诱剂和松饵木在监测松墨天牛及松材     

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.     

Comparison of the potential spread of pinewood nematode (Bursaphelenchus xylophilus) in Finland and Iberia simulated with a cellular automaton model

Pinewood nematode (PWN) is one of the most threatening invasive pests in the pine forests of Europe, and it has recently spread to the Iberian Peninsula via import of timber and wooden packaging material from East Asia. A cellular automaton (CA) model was developed to simulate and compare the potential spread of PWN by transportation and its vectors, Monochamus beetles in the pine forests of Finland and Iberian Peninsula. The model assumes that all pines are equally sensitive to PWN. The CA is a spatio‐temporal grid‐based model, which can easily be applied on different geographical scales. The effects of climate warming and number of entries from ports on the spread of PWN were studied. A sensitivity analysis was conducted on the most uncertain model parameters. Twenty years after hypothetical entries, the predicted area of symptomatic PWN infection (pine wilt disease, PWD) was very low in Finland compared to Iberia. This was because of the low probability of warm July in Finland. The increase in the mean July temperature increased the area of PWD‐infected pine forest relatively more in Finland than in Iberia. An increase in the number of entries also increased the area of PWD‐infected pine forest relatively more in Finland than in Iberia. The probability of PWD infection was the highest in pine forests that were close to entry points and in areas with low elevation and high human population density.     

Comparison of the gene expression profiles of resistant and non‐resistant Japanese black pine inoculated with pine wood nematode using a modified LongSAGE technique

The Japanese black pine (Pinus thunbergii) is highly susceptible to pine wilt disease caused by the pine wood nematode (PWN; Bursaphelenchus xylophilus). To cope with this disease, researchers and tree breeders selected PWN‐resistant individuals in a previous breeding program. In an attempt to understand the mechanisms of resistance in the Japanese black pine, we created four LongSAGE (serial analysis of gene expression) libraries. A total of 20 818 tags were studied, including 5194 tags from a PWN inoculated resistant pine, 5218 a non‐inoculated resistant pine, 5194 an inoculated non‐resistant pine, and 5212 a non‐inoculated non‐resistant pine. The analysis of the libraries indicated that 14 tag species were significantly up‐regulated (e.g., pathogenesis‐related proteins 2 and 4, osmotin, lipoxygenase, and chalcone synthase), and nine were down‐regulated (eukaryotic translation initiation factor SUI1, translationally controlled tumor protein, and xyloglucan endotransglycosylase) by the PWN inoculation in both the resistant and non‐resistant pines. On the other hand, 38 tag species were significantly expressed at a higher level only in the resistant pine (catalase, dienelactone hydrolase family protein) and 25 were expressed at a higher level in the non‐resistant pine (pathogenesis‐related proteins 1, 2, and 3, and leucoanthocyanidin dioxygenase). These differentially expressed genes are presumed to reflect some of the differences between the resistant and non‐resistant pines. Our results provide valuable information on the complex responses induced in the resistant and non‐resistant pine trees in response to PWN invasion.     

<Emphasis Type="Italic">Quercus suber</Emphasis> forest and <Emphasis Type="Italic">Pinus</Emphasis> plantations show different post-fire resilience in Mediterranean north-western Africa

Key message

In the African rim of the Western Mediterranean Basin, cork oak forests and pine plantations coexist. Under similar fire regimes, cork oak forest is more resilient in terms of habitat structure (canopy, understory, and complexity of vegetation strata) than pine plantation. By contrast, both woodland types show similar resilience in plant species composition. Resilience in habitat structure varies between the two woodland types because of the resprouting and seeding strategies of cork oak and pine species, respectively. These differences can be relevant for the conservation of biodiversity of forested ecosystems in a future scenario of increased fire frequency and scale in the Mediterranean basin.

Context

Wildfires have major impacts on ecosystems globally. In fire-prone regions, plant species have developed adaptive traits (resprouting and seeding) to survive and persist due to long evolutionary coexistence with fire. In the African rim of the Western Mediterranean Basin, cork oak forest and pine plantation are the most frequently burnt woodlands. Both species have different strategies to respond fire: cork oak is a resprouter while pines are mostly seeders.

Aims

We have examined the hypothesis that pine plantations are less resilient in habitat structure (canopy, understory, diversity of vegetation strata) and plant composition than cork oak woodlands.

Methods

The habitat structure and plant species composition were measured in 30 burnt and 30 unburnt 700-m transects at 12 burnt sites from north-western Africa, where the two forest types can coexist. Habitat structure and plant species composition were compared between burnt and unburnt transects from cork oak and pine plantation woodlands with generalized linear mixed models and general linear models.

Results

The results showed significant interaction effect of fire and forest type, since cork oak forest was more resilient to fire than was pine plantation in habitat structure. By contrast, both forest types were resilient to fire in the composition of the plant communities, i.e., plant composition prior to fire did not change afterwards.

Conclusion

The higher structural resilience of cork oak forest compared to pine plantation is related to the resprouting and seeding strategies, respectively, of the dominant tree species. Differences in the responses to fire need to be considered in conservation planning for the maintenance of the Mediterranean biodiversity in a future scenario of changes in fire regime.

     

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.