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1.
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. 相似文献
2.
Joung A. Son Masabumi Komatsu Norihisa Matsushita Taizo Hogetsu 《Journal of Forest Research》2010,15(3):186-193
The distribution of pine wood nematodes (Bursaphelenchus xylophilus, PWNs) in Japanese black pine (Pinus thunbergii) tissues was investigated by staining with fluorescein isothiocyanate-conjugated wheat germ agglutinin. After PWNs were inoculated
to current-year stems of pine seedlings, their distribution at about 5 cm below the inoculation site was confined only to
cortical resin canals 1 day after inoculation, and then spread to other tissues, including resin canals of short branches.
When PWNs were inoculated onto cross or tangentially cut surfaces of stem segments, maximal PWN migration speed was estimated
to be faster through cortical resin canals and xylem axial resin canals vertically (>6.7 and <2.3 mm/h, respectively) than
through cortical tissues both vertically and horizontally (<1.2 and <0.2 mm/h). To examine whether PWNs in cortical resin
canals could invade surrounding tissues, segments in which PWNs resided only in cortical resin canals were prepared by removing
the top portion 6 h after inoculation. Additional incubation of such segments caused extended PWN distribution to xylem axial
resin canals and then to other tissues. A similar experiment with top portions of girdled segments removed 12 h after inoculation
also showed extended PWN distribution from xylem axial resin canals and pith to cortical resin canals and then to other tissues.
These results provided direct evidence that PWNs have the ability to migrate from cortical resin canals and xylem axial resin
canals to other tissues. 相似文献
3.
To elucidate the synergetic effects of acid rain on the development of pine wilt disease, we measured the sap flow rate in the stems and the chlorophyll content in the needles of 10‐year‐old Japanese black pine trees, Pinus thunbergii and 12‐year‐old Japanese red pine trees, Pinus densiflora, after exposure to simulated acid rain (SAR, pH 3) or tap water (TW, pH 6.3) as a control. The heat pulse method was used for the estimation of the sap flow rate. No apparent difference was found in the sap flow rate between the trees exposed to SAR and TW, but the chlorophyll content of needles at the end of the treatment was significantly higher in the trees exposed to SAR than in those exposed to TW. When the pinewood nematode, Bursaphelenchus xylophilus, the causal agent of pine wilt disease, was inoculated onto the Japanese black pines that had been exposed to SAR repeatedly for 1 year, the period to death was shortened. Japanese red pines that had been exposed to SAR for 2 years, however, did not show any development of symptoms after the nematode inoculation, suggesting that acid rain only affects pine wilt disease slightly, if at all. 相似文献
4.
The reliability of field, greenhouse and cut‐shoot screening procedures for the assessment of the susceptibility of Scots pine (Pinus sylvestris) genotypes to Melampsora pinitorqua, the causal agent of twisting rust, was evaluated. Fourteen genotypes evaluated on half‐sib progenies after natural infection of 2‐year‐old plants showed comparable rank for rust susceptibility with those evaluated after controlled inoculation of 1‐year‐old seedlings in a greenhouse. The dynamic of pycnia production was assessed at 2‐day intervals in cut‐shoot assays under controlled conditions, giving reliable rankings of Scots pine genotypes between years and being significantly correlated with rust susceptibility assessed after natural infection of 2‐year‐old progenies. These results underlined the importance of some critical factors such as inoculum load and host phenology in the reliability of susceptibility evaluations assessed under different experimental conditions. Artificial inoculation of 1‐year‐old seedlings in greenhouse experiments could provide a useful early test for the management of Scots pine breeding programmes and study of inheritance of twisting rust susceptibility. However, in specific investigations the cut‐shoot assay would constitute a reliable laboratory test for studying host–pathogen interactions and the variability in pathogenicity of Melampsora pinitorqua populations. 相似文献
5.
Rimi Yamaguchi Koji Matsunaga Tomonori Hirao Miho Tamura Atsushi Watanabe 《Forest Pathology》2019,49(4)
Pine wilt disease is of major concern as it has destroyed pine forests in East Asia and Europe. Several studies have suggested that invasion by the pinewood nematode (PWN) Bursaphelenchus xylophilus, which causes this disease, evokes an excessive defence response in pine trees, resulting in tree death. However, few studies have quantitatively evaluated the correlation between PWN distribution and tree defence responses. Therefore, the present study aimed to quantify the number of PWNs and expression levels of putative pathogenesis‐related (PR) genes in different positions of Japanese black pine (Pinus thunbergii) seedlings over time. To quantify the number of PWNs in the seedlings, we used TaqMan quantitative real‐time PCR (qPCR) assay. During the early phase of infection, most PWNs were distributed around the inoculated sites, with only a small number being detected at distant sites, but the expression levels of PR genes were highly upregulated throughout the seedlings. Both the number of PWNs and expression levels of PR genes then increased drastically throughout the seedlings, all of which exhibited external symptoms. Thus, it appears that the rapid migration of PWNs induces a defence response throughout the seedling; however, this may not be effective in controlling these parasites, thereby ultimately leading to plant death. 相似文献
6.
Simulated acid rain (SAR) at three pH levels (pH 4, 3, 2) was applied to only the top or both the top and roots of 4-month-old
Japanese black pine seedlings repeatedly for two months. Then the seedlings were inoculated with a virulent isolate (S10)
of pinewood nematode,Bursaphelenchus xylophilus. The exposure to SAR at any level did not kill the seedlings. When inoculated with nematodes, however, the nematode population
build-up was significantly higher in the seedlings pretreated with SAR at any level than in the control at the 7th and 17th
day after inoculation, and subsequent disease symptom development was also significantly accelerated by the exposure to SAR
at pH 2 and 3. This result indicates that even acid rain at pH 4 has the potential for promoting population growth of pinewood
nematodes in 4-month-old Japanese black pine seedlings. 相似文献
7.
R.‐L. Petäistö 《Forest Pathology》2008,38(1):1-15
First‐ and second‐year containerized Norway spruce seedlings were inoculated with conidia of type A (large tree type) and type B (small tree type) of Gremmeniella abietina var. abietina at different times during the summer. The appearance of symptoms after artificial inoculation and natural infection on spruce seedlings were recorded the following spring and compared with the disease symptoms on Scots pine seedlings. The proportion of diseased seedlings after inoculation reached as high as 80%. The susceptible period during the summer began later on the first‐year seedlings than on the second‐year seedlings, and was similar for the pine seedlings. Susceptibility of first‐year seedlings was highest in August and on second‐year seedlings in July. The accumulated temperature sum, relative humidity and height growth for first‐ and second‐year seedlings was assessed. Natural infection in 2002 caused more disease on pine than on spruce seedlings. Experimental thinning of seedlings had no effect on disease incidence. In a preliminary comparison between the ability of A and B types to cause disease in Norway spruce seedlings, type B caused more damage than type A after inoculation. However, type A caused a high disease frequency in other experiments in this study. Symptoms on Norway spruce seedlings often first occurred in the mid‐section of the shoot, and were similar to those observed on pine seedlings: needles turned brown, starting at the needle base, in the spring following inoculation. On first‐year spruce, diseased needles were shed rapidly, in contrast to a slower rate of shedding on first‐year pine seedlings. Pycnidia developed about 2 years after inoculation (on pine 1 year after inoculation). On Norway spruce seedlings the lower part of the shoot, including the lateral shoots, often remained alive. The experiments show that G. abietina can cause disease on containerized Norway spruce seedlings under nursery conditions in Finland. The coincidence of spore dispersal, seedling susceptibility and predisposing factors are important in disease development. 相似文献
8.
T. Regliński N. Rodenburg J. T. Taylor G. L. Northcott A.
Ah
Chee T. M. Spiers R. A. Hill 《Forest Pathology》2012,42(1):75-78
Root drench application of Trichoderma atroviride isolates R32, R33, R40 and R84 promoted the growth of potted radiata pine seedlings. After 6 weeks, seedlings treated with R33 and R84 had thicker stems and greater stem and root biomass (p < 0.05) than untreated controls. Treatment with R32 increased seedling root biomass whilst R40 increased stem diameter. None of the isolates affected seedling height. One isolate, R33, induced systemic resistance to stem inoculation with Diplodia pinea and reduced dieback incidence by 20% compared with untreated controls. To our knowledge, this is the first report of systemic induced resistance by Trichoderma in a pine species. Furthermore, seedlings that were treated with R33 (root drench) plus foliar application of methyl jasmonate (MeJA) expressed elevated peroxidase activity in their stems 2 weeks later, compared with seedlings treated only with MeJA. Because R33 itself did not affect peroxidase activity, this may be indicative of treatment synergy or defence potentiation by R33. Curiously, R33 + MeJA induced terpenoids but suppressed phenylalanine ammonia‐lyase activity suggesting possible trade‐offs between phenolic and terpenoid defence pathways in the treated seedlings. 相似文献
9.
Migration of Bursaphelenchus xylophilus in cortical and xylem axial resin canals of resistant pines
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Migration of the pine wood nematode (PWN), Bursaphelenchus xylophilus, in susceptible and resistant pines was investigated at the tissue level. PWN was inoculated onto the top cross‐cut surface of 20‐cm stem cuttings of susceptible Pinus thunbergii and resistant pines (P. strobus, P. rigida and P. thunbergii of a resistant family Namikata‐(t)‐73 (half‐sib)). PWNs were mainly distributed in cortical resin canals of susceptible P. thunbergii down to 15 cm from the inoculated surface by 6 h after inoculation (HAI) and all tissues (including cortical and xylem resin canals) down to the bottom at 192 HAI. In P. strobus, P. rigida and P. thunbergii family Namikata‐(t)‐73 (half‐sib), PWN was distributed in cortical resin canals down to 5 cm by 6 HAI and down to the base at 192 HAI. However, the distribution of PWN in xylem resin canals of the resistant pines was restricted near inoculated surfaces down to 5 cm, even at 192 HAI. These results demonstrated that migration of PWN in resistant pines was slowed in cortical resin canals and restricted in xylem axial resin canals, features which may be associated with the resistance. 相似文献
10.
In order to study the changes in ectomycorrhizal development during symptom expression of pine wilt disease, root window observations were conducted concurrent with measurements of leaf water potential as well as photosynthetic and transpiration rates of 5‐year‐old Pinus thunbergii trees that were inoculated with the pinewood nematode (PWN) Bursaphelenchus xylophilus. Infected trees were compared with girdled and uninfected control trees. Ectomycorrhizas developed constantly during the experimental period in control trees but did not develop in the girdled trees. Ectomycorrhizal development ceased within 2 weeks in those trees that finally died after PWN infection. In the trees that survived PWN infection, ectomycorrhizal development ceased within 1–4 weeks of inoculation but was resumed thereafter within 3–6 weeks. Ectomycorrhizal development ceased prior to a decrease in both photosynthetic rate and leaf water potential in the inoculated trees. 相似文献
11.
Masabumi Komatsu Jounga Son Norihisa Matsushita Taizo Hogetsu 《Journal of Forest Research》2008,13(2):132-136
Pine wilt disease, caused by the pine wood nematode (PWN, Bursaphelenchus xylophilus), is a major threat to pine forests throughout East Asia. Nonetheless, its mechanism of invasion has not yet been described
in detail. To better understand the pathology of this disease, it is important to examine the distribution of PWNs within
pine tissue during the course of disease development. We attempted to stain nematodes with fluorescein-conjugated wheat germ
agglutinin (F-WGA) as a means to locate and track the spread of PWNs. Although PWNs proliferated on Botrytis cinerea fungus were successfully stained only on their vulvas and spicule holes, PWNs extracted from inoculated Pinus thunbergii seedlings were stained on their surface. Stainability, or the percentage of PWNs stained with F-WGA over more than half of
their surface, was about 20% by 1 day after inoculation, but increased to 80% at 10 days. The stainability of PWNs extracted
from a 5-cm main stem segment that included the inoculation site was less than that of PWNs extracted from other parts of
the main stem farther away (i.e., those that had dispersed). These results suggest that stainability is related to dispersal
activity in time. Thus, to raise the stainability of PWNs at shorter timeframes after inoculation, PWNs with more than 80%
stainability were re-inoculated into pine seedlings. This resulted in more than 70% stainability from 1 to 6 days after inoculation.
In F-WGA stained thin paraffin sections of pine tissue of re-inoculated seedlings, PWNs brightly fluoresced under epifluorescence
and were easily detected against the dark background of pine tissue. This staining technique with F-WGA is an excellent tool
for detecting PWNs in pine tissue. 相似文献
12.
Enhancing resistance in Pinus radiata seedlings to terminal crook (Colletotrichum acutatum) using methyl jasmonate and ultraviolet‐C radiation
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Colletotrichum acutatum is a fungal pathogen that causes terminal crook disease in radiata pine (Pinus radiata) seedlings in New Zealand forest nurseries. Symptoms of infection include malformation or death of the growing tip and a stiffening and thickening of the stem. Although the disease can be managed effectively using fungicides, the New Zealand forest industry is interested in alternative control options such as induced resistance. The aim of this study was to evaluate spray application of chitosan (1.4 g/l) or 2.25 mm methyl jasmonate (MeJA) and irradiation with UV‐C (2.16 kJ/m2) for their potential to induce resistance to terminal crook. The treatments were applied to 4‐month‐old seedlings at 1 week before pathogen inoculation. By the end of the experimental period (42 days after inoculation), there was 80% disease incidence in the controls, with 48% of seedlings exhibiting severe terminal crook symptoms. The most effective treatment (p < 0.05) was MeJA with 16% disease incidence and none with severe symptoms. UV‐C also significantly (p < 0.05) reduced infection with 52% incidence and 20% of seedlings exhibiting severe symptoms. Chitosan did not reduce disease incidence (72%) compared with the control (80%) but did significantly reduce (p < 0.05) disease severity with 28% exhibiting severe symptoms. MeJA was the only treatment that significantly (p < 0.05) reduced the detrimental effects of infection on seedling apical growth and stem diameter. To our knowledge, this is the first report demonstrating the potential for MeJA and UV‐C to control terminal crook in radiata pine. 相似文献
13.
Mats Hannerz ke Thorsn Staffan Mattsson Jan Weslien 《Forest Ecology and Management》2002,160(1-3):11-17
Damage caused by pine weevil (Hylobius abietus L.) to planted seedlings and cuttings of Norway spruce (Picea abies (L.) Karst.) was studied at five clearcut sites in south-eastern Sweden. The main objective was to compare the two types of stock in terms of attack frequency and mortality due to pine weevil feeding. Cuttings and seedlings with the same initial stem-base diameter (4 mm) were compared. Two sites were harvested and scarified shortly before planting, two were harvested shortly before planting, but were not scarified, and one was harvested 2 years before and scarified the autumn before planting. The total mortality 5 years after planting was highest, greater than 90%, at the new, non-scarified sites, and lowest, 23%, at the old, scarified site. More than 90% of the mortality was caused by pine weevil feeding. Attack frequency and pine weevil induced mortality were significantly higher among seedlings than among cuttings. Mortality due to pine weevil damage was 4–43% higher in seedlings than in cuttings after the fifth year. Of the cuttings and seedlings that were attacked in the first year, a significantly higher frequency of the seedlings were girdled. The higher resistance of cuttings to pine weevil damage may partly explain the more rapid growth of cuttings reported in other studies. However, the causes of their higher resistance need to be further investigated. The thicker bark and needles on the stem base of the cuttings could be important in this respect. 相似文献
14.
The height growth of Korean pine plantation was investigated for different ages of saplings in Benxi city of the eastern Liaoning, China. The sapling and seedling age were determined by branch annuals. Results show that the mean height of third- to eight-year Korean pines ranges from 13.31 cm to 111.73 cm and only 19.4% of the saplings reached the height above 130 cm. The height (Y) of eight-year-old Korean pine increases linearly with the increment of ground stem (x) (Y=3.1x+35.15). The relation between height and age can be described by an exponential equation. The second year after planting of the Korean pine is considered as the key period of Korean pine growth really adapting to the local environment; plantation management should be strengthened in this year. The Korean pine seedlings need two years to adapt to the local environment, thus early care and management should be taken in the second year. 相似文献
15.
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。 相似文献
16.
Tolerance of bareroot and container-grown seedlings of black spruce (Picea mariana (Mill.) B.S.P.), jack pine (Pinus banksiana Lamb.), and eastern white pine (Pinus strobus L.) to competition from herbaceous vegetation was examined in the first five years after planting on a site in the Great Lakes/St. Lawrence forest of Ontario, Canada. Shoot and root morphological characteristics of various stocktypes were measured before planting and correlated with 5-year survival and growth following control and no control of herbaceous vegetation. For black spruce and jack pine, medium-sized bareroot stocktypes had greater relative 5-year stem volume growth in the presence of herbaceous vegetation than did container stock of either species or large bareroot stock of spruce. Relative volume growth was measured as the ratio of the cumulative stem volume increment in the presence of vegetation (Veg) to that in the absence of vegetation (NoVeg), i.e., the Veg:NoVeg ratio. In white pine, the Veg:NoVeg ratio of volume increment of medium container and large bareroot stocktypes exceeded that of small container and medium bareroot stocktypes. In jack pine, root collar diameter at planting and number of first-order lateral roots were positively correlated with 5-year Veg:NoVeg ratio of volume increment. In white pine, the Veg:NoVeg ratio was also positively correlated with root collar diameter at planting and with root volume. In black spruce, the ratio was not related to pre-plant morphology. Thus, for white pine and jack pine, certain pre-plant morphological features may be useful in forecasting the relative ability of different stocktypes to grow under herbaceous competition conditions in the field. 相似文献
17.
Takeshi Taniguchi Chihiro Tanaka Shigenobu Tamai Norikazu Yamanaka Kazuyoshi Futai 《Journal of Forest Research》2008,13(4):233-240
The regeneration of Japanese black pine (Pinus thunbergii) seedlings is inhibited in a black locust (Robinia pseudoacacia)-dominated area. We examined the presence of pathogenic fungi in Japanese black pine seedlings in the area in order to determine
the effect of pathogenic fungi on the inhibition of regeneration. When Japanese black pine seedlings were planted in the soil
obtained from a black locust-dominated area, all of the seedlings died under low-intensity light conditions, whereas 84% of
the seedlings survived in the soil obtained from a Japanese black pine-dominated area under the same light conditions. One
fungus was isolated from 48.7% of the dead pine seedlings and was identified as Cylindrocladium pacificum Kang, Crous & Schoch, based on the morphological characteristics, growth, and DNA analysis. This fungus was also isolated
from 50% of the dead pine seedlings in 2005 and 66.7% of the seedlings in 2006—both were planted in a black locust-dominated
area. The virulence of this fungus increased under high-nitrogen and/or low-intensity light conditions. These results reveal
the possibility that the soil eutrophication and shading by the black locust are conducive to a severe damping-off disease
and threaten the survival and regeneration of Japanese black pine seedlings. 相似文献
18.
The aim of this study was to test the pathogenicity of two Spanish isolates of Heterobasidion annosum sensu stricto in 2‐year‐old Pinus pinaster seedlings. Two types of inocula (woodchips and sawdust) were used to infect the seedlings by two different routes (stem inoculation and soil infestation). The mortality rates of the stem‐inoculated seedlings differed significantly from controls, but those of the seedlings infected via soil infestation did not differ. For both types of inoculation, the lesions were longer, and wilting symptoms were more severe in the seedlings inoculated with H. annosum than in control seedlings. For stem inoculation, biomass allocation did not differ significantly between the infected and control seedlings. However, the percentage of fine roots was lower in seedlings infected via soil infestation than in the control seedlings. To our knowledge, this is the first pathogenicity test with H. annosum isolates and P. pinaster. 相似文献
19.
In order to assess the influence of water stress on the development of Sphaeropsis sapinea cankers in Pinus halepensis, the stems of 4‐ to 5‐year‐old potted seedlings were artificially inoculated with the fungus before and after being kept at controlled water regimes from April 1997 to March 1998. In the pre‐water‐stress inoculation experiment, the canker length, measured 5 months after inoculation (September 1997), was greater in seedlings predisposed to extreme water deficit (midday needle water potential between ?4.5 and ?5.5 MPa). In the post‐water‐stress inoculation experiment, the fungus was inoculated in April 1998, after irrigation had enabled the seedlings to resume normal needle water potential. In this case also, at 5 months after inoculation, longer cankers were visible in seedlings that had been subjected to extreme water deficit. These findings suggest that the occurrence of marked water stress, although apparently tolerated by Aleppo pine, can enhance the development of S. sapinea cankers in this species, regardless of whether the stress occurs before or after infection by the fungus. 相似文献
20.
Six‐week‐old seedlings of slash pine (Pinus elliottii) were inoculated with sporidia of Cronartium strobilinum, which is the cause of southern cone rust disease, by wounding the apical tips of the seedlings. Pycnial sori developed abundantly within 1 month of inoculation on the epicotyl and primary needles. The progress of the pathogen was confirmed histologically within the cells of the stem and haustoria were observed near the nucleus of the cells. In the needles, pycnial pustules developed and the fungus was abundant in the mesophyll. 相似文献