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1.
The fungal pathogen Septoria musiva can be difficult to isolate from cankers that result from its colonization of poplar stems, and its persistence in these cankers has not been well studied. In order to compare cultural and polymerase chain reaction (PCR)‐based assays for detection of S. musiva in cankers, stems of susceptible hybrid poplar clone NC11505 were wounded and inoculated in August 2003. At 8, 16, 24 and 32 weeks after inoculation (October and December 2003, February and April 2004, respectively), 110 inoculated stems (plus controls) were harvested and a semiselective culture medium was used in attempts to detect the pathogen in bark and wood. Six chips of bark and six chips of underlying wood from one half of each canker were incubated on the semiselective medium for 2 weeks until pycnidia and conidia of S. musiva could be identified. The number of positive cankers and positive chips (out of six attempts per tissue per canker) was recorded. The remaining halves of cankers from subsets of 70 inoculated stems (plus controls) of those harvested in October 2003 and April 2004 were tested using a PCR‐based assay. Three chips of bark and three chips of underlying wood were ground, and DNA was extracted and then amplified using S. musiva‐specific primers designed from the internal transcribed spacer (ITS) region of nuclear rDNA repeats. The number of positive cankers and positive chips (out of three attempts per tissue per canker) was recorded. For both assays, the number of positive cankers and the number of positive chips per canker decreased with time. Using either assay, however, the pathogen was still detected from at least 49% of cankers at 32 weeks after inoculation.  相似文献   

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Hymenoscyphus fraxineus, the causal agent of ash dieback, was inoculated onto intact, unwounded current‐year shoots and leaf scars of 4‐year‐old, potted Fraxinus excelsior seedlings. Pieces of ash wood colonized by the fungus were used as inoculum. Three of 25 (12%) of the inoculated intact shoots and nine of 25 (36%) of the inoculated leaf scars were infected by H. fraxineus and developed typical symptoms of ash dieback, including necrotic lesions on the shoot surface and wood discoloration as well as shoot and leaf wilting distal to the inoculation site. No symptoms occurred on control seedlings, which had been inoculated in the same way but with sterile wood pieces. Visible necrotic lesions on shoots and wood discoloration were statistically significantly longer in proximal than in distal direction from the inoculation site, a pattern which resembles symptoms after natural infection. The ash dieback pathogen was re‐isolated from nine of 12 (75%) of the symptomatic seedlings. These results provide indirect supportive evidence that the fungus infects shoots via leaves and shows that it is able, under experimental conditions using a massive mycelial inoculum, to directly infect intact, unwounded current‐year shoots of its main host in Europe.  相似文献   

5.
Mass mortality of oak trees has been occurring in Japan since the late 1980s. The fungus Raffaelea quercivora has been frequently isolated from discoloured sapwood in dead or wilting trees and inoculation experiments have shown it to be capable of causing wilting and xylem discoloration in several oak species, notably Quercus crispula and Q. serrata. In this study, we inoculated seedlings of six Fagaceae species with R. quercivora and, after 56 days, measured the vertical length of the discoloration and the areas of discoloured and non‐conducting sapwood on stem cross‐sections. The sapwood discoloration and the water non‐conduction areas were larger in Q. crispula and Q. serrata than in the other species.  相似文献   

6.
Five conifer species grown in the Great Lakes region of North America were examined for their susceptibility to Fusarium circinatum, (syns. Fusarium subglutinans f. sp. pini and F. moniliforme var. subglutinans), the causal agent of pitch canker. Three‐year‐old (3‐0) seedlings of red (Pinus resinosa), jack (P. banksiana) eastern white (P. strobus), Scots (P. sylvestris) and Austrian (P. nigra) pine were planted in 4 l pots in a greenhouse at Auburn University in November 1998. In April and June 1999, seedlings were inoculated by removing a needle fascicle approximately 5 cm from the terminal bud and placing a drop containing F. circinatum conidia on the wound. Resin production, canker length and seedling mortality were recorded 12 weeks later. Jack, Scots and eastern white pine were the most susceptible with Austrian and red pine more resistant to the fungus. F. circinatum was re‐isolated from 37% to 96% of inoculated seedlings. The susceptibility of jack, Scots and eastern white pine indicates a potential risk to these important species of the region if F. circinatum were to be introduced into the area.  相似文献   

7.
In this study a Sphaeropsis sp. is recorded as causing cankers on cypress in South Africa. These cankers are either found alone or on trees which are infected with the well-known cypress canker pathogen, Seiridium unicorne. When the two fungi occurred together, lesions could not be distinguished from each other, although the greater number of lesions were caused by the Sphaeropsis sp. The fungus appears to be identical to that described as Sphaeropsis sapinea f.sp. cupressi, in Israel, which is a fungus very different from the pine pathogen, Sphaeropsis sapinea. Pathogenicity tests showed that the Sphaeropsis sp. is significantly more pathogenic to Cupressus lusitanica than to Pinus roxburgii and Pinus elliottii. S. sapinea was pathogenic only to the Pinus spp. tested, and not to C. lusitanica. In contrast, the cypress pathogen Seiridium unicorne, was pathogenic to C. lusitanica and also to the two Pinus spp. tested.  相似文献   

8.
Phenylalanine ammonia lyase (PAL) activity increased rapidly in Pinus radiata seedlings after treatment with 2 mm 5-chlorosalicylic acid (5CSA) and peaked after 48 h at four-times the level of the control. Elevation of PAF is often associated with the induction of host resistance mechanisms. Thus, effects of 5CSA on subsequent infection of P. radiata by Sphaeropsis sapinea was investigated. Only one out of 16 seedlings treated with 2 mm 5CSA, 4 days before inoculation, developed deadtop compared to 14 of the controls. The 5CSA was directly inhibitory at this concentration and reduced growth rate of S. sapinea on agar by 27%. However, antifungal activity does not fully account for the observed disease control afforded by 5CSA since 0.5 mm and 0.25 mm 5CSA, which did not inhibit growth of S. sapinea, significantly reduced the incidence of deadtop. Furthermore, 1 mm 5CSA protected seedlings against fresh inoculation challenge for up to 32 days after application. Such durable resistance is unlikely to be attributable to the direct effects of the applied compound and may suggest the involvement of inducible host resistance mechanisms.  相似文献   

9.
Open-pollinated progenies from two trembling aspen (Populus tremuloides) families with a 30-year field history of 91% (putatively susceptible, PS) and 45% (putatively resistant, PR) mortality, respectively, caused by Hypoxylon mammatum, were grown in a greenhouse for 3 and 9 months. Seedlings were inoculated with two isolates of H. mammatum, representing high and low aggressiveness, through a stem wound. Beginning 4 days after inoculation, host responses were examined on 312 seedlings per family for a period of 5 weeks on 3- and 9-month-old seedlings. Significant differences in host responses were observed between the two isolates, with the more aggressive isolate allowing less callus formation, causing more tissue necrosis and producing longer cankers than the less aggressive isolate on both aspen families. Between-family difference was much greater than within-family difference for tissue necrosis, callus formation and canker length for both seedling ages. When the 3-month-old seedlings were challenged with H. mammatum, the PR family showed greater tissue necrosis, longer cankers, and less callus formation than the PS family. However, when the more lignifìed 9-month-old seedlings of the same families were inoculated, the PR seedlings had much less tissue necrosis, shorter cankers, more callus formation and less seedling mortality than the PS family. The results indicate that artificial inoculation of seedlings in a greenhouse can differentiate two aspen families at the seedling stage, but their correlation to field performance is highly dependent upon the seedling age. Host responses of older seedlings may be a better indicator of field performance.  相似文献   

10.
Research on tree response to pest attacks under drought and heat stress is crucial for predicting the effect of climate change on forest pests and tree health. In this work, we studied the influence of severe water stress on the resistance of Pinus yunnanensis to inoculations of Grosmannia yunnanensis, a pathogenic fungus associated with the aggressive bark beetle, Tomicus yunnanensis. Experiments were performed in two plots, one located at the top of a hill and the other at the foot of a hill, in Xiaojiu, Yunnan, China, from May to September 2010 and March to April 2012. Measurements of soil and needle water content and predawn needle water potentials confirmed that severe droughts occurred during these periods. Maximum water stress intensity even occurred in 2010 during the typical rainy season. Following low‐density inoculations, the reaction zone length and fungal growth were measured in the phloem. Sapwood blue staining and total sapwood occlusion were quantified following fungal mass inoculations. Our results suggest that severe water stress has a negative impact on a tree's resistance to G. yunnanensis. By comparing our results to previous work performed from 1997 to 1999 on the same tree species, and in the same locality under mild water stress, we concluded that drought had opposite effects on tree resistance above and below a certain threshold of predawn needle water potential. The threshold, corresponded to maximum efficiency for tree defence mechanisms, was visible in our results, and was estimated to be approximately ‐1.1 MPa to ‐1.2 MPa for P. yunnanensis under local conditions. We suggest that the existence of such a threshold of water stress intensity may be viable for any conifer undergoing biotic aggressions and, specifically, bark beetle attacks.  相似文献   

11.
The pathogen Sphaeropsis sapinea can persist in stems and branches of asymptomatic pines and can later induce disease when triggered by host stress. Several experiments were conducted to test if:(i) medium amended with tannic acid (TA) can increase the frequency of cultural detection of this shoot blight and canker pathogen from asymptomatic red pine (Pinus resinosa) stems, and (ii) S. sapinea can persist in asymptomatic red pine in the field following artificial inoculation. TA (0.5% w/v) in 2% (w/v) water agar proved to be the best medium for isolation of S. sapinea among a larger number of tested media. The addition of TA had little or no effect on the growth of two group A and two group B isolates of S. sapinea. However, when TA was added, 11 other fast‐growing fungal isolates from stems/branches of red or jack pines (P. banksiana) were inhibited and grew more slowly (p < 0.05) than both S. sapinea groups. The TA‐amended medium improved cultural detection of S. sapinea from 2‐year‐old, asymptomatic red pine nursery seedlings compared with two other methods used for the cultural detection of S. sapinea (32%vs. 8.5% and 18% recovery; p < 0.001 and p = 0.031, respectively). A field test using the TA‐amended media established that S. sapinea can persist asymptomatically in red pine trees for at least 1 year. This medium significantly reduces the frequency of false‐negatives from asymptomatic field material.  相似文献   

12.
Roberts  J.J.  Zwiazek  J.J. 《New Forests》1999,18(3):301-314
Five week old white spruce [Picea glauca (Moench) Voss] seedlings were placed within controlled-temperature growth chambers, and subjected to one of four periodic, chilling treatments for approximately 16 weeks. The treatments differed in the frequency of 24 hour exposure to 5 °C the seedlings received, and ranged from no chilling (control), to once every two, three or four days. After 25 weeks of growth, the control seedlings were significantly taller than all chilling-exposed seedlings, but stem diameters were similar. Chilling did not affect either needle length or needle density compared to control seedlings. During a water deficit test, frequently chilling-exposed seedlings maintained significantly higher mid-day shoot water potentials under extreme water stress. Re-watering after the drought resulted in all chilling-treated seedlings recovering faster to pre-stress shoot water potential levels than the control seedlings. Measurements of new root growth after 21 days at 10 °C indicated that previous chilling exposure delayed or inhibited new root production.  相似文献   

13.
The effects of water stress and defoliation by Lymantria dispar on the successful infection of the fungal pathogen Biscogniauxia mediterranea in seedlings of the Turkey oak (Quercus cerris) were studied under controlled conditions. Seedlings were inoculated by spraying with a spore suspension of Biscogniauxia mediterranea and were subjected to different levels of water stress, insect defoliation, and combinations of the two factors. The leaf water potential was periodically assessed throughout the experiment. The fungus was isolated exclusively from seedlings deprived of water, and insect defoliation increased significantly the proportion of fungus isolation when the water stress was moderate. Thus insect defoliation seems to promote fungal colonization in case of moderate water stress.  相似文献   

14.
We investigated variation in virulence of Cryphonectria hypovirus 1 (CHV‐1) to the chestnut blight fungus, Cryphonectria parasitica, in Macedonia by inoculating chestnut stems in the field. We inoculated trees with two isolates of C. parasitica, each infected with one of five isolates of CHV‐1, four of which were the same for both fungal isolates. Two virus isolates, [Sk28] and [Sk47], were significantly more virulent than the others when compared in the same fungal host isolates, as measured by reduced canker growth and increased callus formation. Mycelial growth rate in vitro was weakly correlated to canker growth or callus formation and is therefore not a reliable predictor for virulence. We found significant fungus × virus interactions for canker growth and callus formation, which seems due mainly to one virus isolate. Significant interactions were not expected because the two fungal host isolates are members of the same clone that is dominant in Macedonia and most of southeastern Europe. Phenotypic variation for response to viruses, therefore, is greater than variation revealed by the genetic markers used to define clones. More than half of the trees inoculated with virus‐free controls were dead within 2 years, and the 30% still alive after 5 years had cankers with extensive callus formation, indicating that natural virus transmission had occurred after inoculation. In contrast, only 2% of the trees inoculated with virus‐infected isolates were dead after 5 years. Hypoviruses naturally occurring in Macedonia reduce canker development and tree mortality similarly to those in other parts of southern Europe, and therefore, may have good potential for biological control of chestnut blight.  相似文献   

15.
The aim was to determine the inoculation density above which Scots pine (Pinus sylvestris) is overcome by the blue‐stain fungus Ophiostoma ips that is associated with the bark beetle Ips sexdentatus. In north‐western Spain, stems of 16 Scots pines were inoculated at various densities (0, 400, 800 or 1600 inoculi/m2) along circumferential 100 or 150 cm wide inoculation belts. Each inoculum consisted of a 5 mm diameter cylinder of malt extract agar colonized by the fungus. Three months later, all trees were harvested and trunk resinosis and foliage colour were visually assessed. The percentage of healthy, desiccated, resin soaked, and blue‐stained sapwood, as well as growth productivity indices, were calculated from stem disks cut from within the inoculated zone of each tree. Sapwood‐specific hydraulic conductivity (Ks) of each tree was measured in the middle of the inoculated zone. All parameters of tree vigour changed dramatically to the worse when inoculation densities were above 400 inoculi/m2, and foliage changed from green to yellow‐green or yellow when an inoculation density of 800 instead of 400 was used. The percentage loss of sapwood‐specific conductivity (PLC) increased from 30 to 90% and the percentage of healthy, conductive sapwood dropped from 85 to 35% at 800 inoculi/m2. No effect of the width of the inoculation belt was observed, and there was no relationship between tree productivity indices and the level of resistance. A non‐linear negative relationship was found between PLC and the percentage of healthy sapwood. It is concluded that tree resistance was overcome and that trees were going to die when the inoculation density was ≥800 inoculi/m2.  相似文献   

16.
A new disease, causing mortality of Italian cypress (Cupressus sempervirens) trees cv. ‘Totem', was observed in an Israeli nursery. Fusarium compactum was isolated from cankers, and its pathogenicity was confirmed by Koch's test. The fungus readily formed conidia and chlamydospores in PDA culture. Growth of F. compactum was fastest at 26–30°C. Artificial inoculations with F. compactum caused severe cankering of several species of each of the following coniferous genera: Cupressus, Juniperus, Tetraclinis, Thuja, Cedrus and Pinus. Canker development was significantly enhanced when extreme water stress was imposed on cypress saplings prior to inoculation.  相似文献   

17.
Climate change poses severe pressures to European conifer forests. Using non-native tree species, such as Douglas fir (Pseudotsuga menziesii), is one proposed strategy to circumvent adverse effects for forest management. However, novel forest health risks can impair the cultivation of non-native trees. In 2022, we observed large Douglas fir trees (approximately 40–50 years old, diameter at breast height (dbh) 21–41 cm) that had recently died in spring or summer 2022 in three forest stands in Eastern Austria. Intensive resin flow, blue-staining of the sapwood and the absence of bark- and wood-boring insects indicated a fungal infection. Isolations from blue-stained sapwood of the dead trees consistently yielded cultures of the opportunistic pathogen Diplodia sapinea. In a greenhouse wound inoculation experiment, seven D. sapinea isolates obtained from Douglas fir caused phloem necrosis, blue-staining of sapwood and mortality and thus displayed pathogenicity towards seedlings of both Ps. menziesii and its common host, Scots pine (Pinus sylvestris). Diplodia sapinea produced significantly longer areas of blue-stain as well as higher and faster mortality in Ps. menziesii compared to P. sylvestris. We conclude that D. sapinea substantially contributed to the death of seven of the 13 examined large Douglas fir trees. While this fungus has been described as a pathogen of young Douglas fir trees before, this is the first report that it can potentially kill large individuals of this conifer species under drought conditions. Thus, our results indicate that D. sapinea could represent a severe threat to the cultivation of Ps. menziesii in European forestry.  相似文献   

18.
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   

19.
A series of experiments was conducted to determine the relative tolerance in vitro of an isolate of Septoria musiva (a fungus that causes a severely damaging stem canker disease of poplars) for selected chemicals. Inhibition of diameter growth of this fungus on a V‐8 vegetable juice‐based medium with captan, chlorothalonil, iprodione, mancozeb and streptomycin sulphate at concentrations, respectively, of 50, 1, 10, 10, and 100 mg l–1 was relatively low compared to inhibition of eight other fungi cultured from cankers on poplars. In addition, the presence of captan stimulated profuse sporulation of the fungus. These properties assisted in the identification of S. musiva from cankers resulting from artificial inoculation of poplar branches in the field.  相似文献   

20.
Ramets of the Cupressus sempervirens clone Bolgheri were raised in three CO2 concentrations in a naturally CO2‐enriched environment [360 (control), 680 and 1670 μmol mol–1 CO2]. Bolgheri is patented as resistant to Seiridium cardinale. The ramets were artificially inoculated using a virulent isolate of S. cardinale. Healing of the resulting cankers was visually assessed over 6 years. Six months after inoculation, cankers on trees grown at 680 μmol mol–1 were significantly larger than the controls. However, no significant trend was recorded at later dates. In conclusion, the Bolgheri clone maintained its resistance to S. cardinale and could be used to control cypress canker even in increasing atmospheric CO2 concentrations.  相似文献   

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