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1.
Oak decline has been a serious problem in Europe since the beginning of the twentieth century. In south‐west Spain, Quercus ilex and Q. suber are the main affected species, and their decline has been associated with Phytophthora cinnamomi. During the last 10 years, a severe decline of Q. ilex and Q. faginea accompanied by a significant decrease in the production of acorns affecting natural regeneration was observed in the eastern part of the Iberian Peninsula. Therefore, the aim of this study was to investigate the possible involvement of Phytophthora spp. in the decline. A forest in the Natural Park ‘Carrascar de la Font Roja’ in Comunidad Valenciana (eastern Spain), which is dominated by Q. ilex and Q. faginea, was surveyed during 2010–2011. Symptomatic trees showed thinning and dieback of the crown, withering of leaves and death. An extensive loss of both lateral small woody roots and fine roots and callusing or open cankers on suberized roots were observed. Soil samples containing fine roots were baited using both Q. robur leaves and apple fruits. Six Phytophthora species were isolated: P. cryptogea, P. gonapodyides, P. megasperma, P. quercina, P. psychrophila and P. syringae. These are the first records of P. quercina and P. psychrophila on Q. faginea, of P. quercina in Spain and of P. psychrophila in mainland Spain. A soil infestation trial was conducted for 6 months under controlled conditions with 1‐year‐old seedlings of Q. ilex and Q. faginea. Phytophthora cinnamomi was included in the pathogenicity test for comparison. The results showed that Q. ilex seedlings were generally more susceptible to infection than Q. faginea with P. cinnamomi being the most aggressive pathogen to both oak species. The two most commonly isolated Phytophthora species, P. quercina and P. psychrophila, also proved their pathogenicity towards both Q. ilex and Q. faginea.  相似文献   

2.
The formation of mycorrhiza on Pinus tabulaeformis (Pt) seedlings by ectomycorrhizal fungi (ECMF), the protective effect of the ECMF on damping‐off caused by Rhizoctonia solani and the expression of pathogenesis‐related proteins (PR‐proteins) induced by Amanita vaginata and/or R. solani in Pt seedlings were investigated. The results indicated that: (i) among the 10 ECMF tested, five species (Gomphidius viscidus, Suillus laricinus, Suillus tomentosus, A. vaginata and Basidiomycetes no. 058) formed mycorrhiza with Pt seedlings. There was a positive correlation between mycorrhizal colonization levels and protective effects (r = 0.9207). Among the five species, A. vaginata had the highest colonization level and the best protective effect against damping‐off; however, G. viscidus had no protective effect even though it had a high mycorrhization level. (ii) Chitinases, β‐1,3‐glucanases and PR‐proteins in the seedlings were not induced by A. vaginata colonization, but were induced when A. vaginata‐mycorrhizal seedlings were attacked by R. solani. It is possible that chitinases, β‐1,3‐glucanases and 108, 97, 78, 44, 37 and 14‐kDa PR‐proteins induced both by A. vaginata and R. solani protect Pt seedlings from R. solani infection.  相似文献   

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Between 2013 and 2015, several surveys were conducted throughout forest areas in the north of Iran with the aim of isolating and identifying fungi present on trees showing decline symptoms. Fungal species, isolated from symptomatic wood tissues of various tree genera, including Zelkova, Acer, Parrotia, Quercus, Diospyros, Fagus, Fraxinus, Populus, Pinus, Pterocarya, Gleditsia, Carpinus, Ulmus, Crataegus, Alnus, Mespilus, Punica, Cydonia, Cupressus and Prunus, were morphologically identified and characterized. A molecular identification based on comparisons of DNA sequences of the β‐tubulin gene allowed us to identify seven species belonging to Phaeoacremonium; namely, Phaeoacremonium parasiticum, Pm. alvesii, Pm. minimum, Pm. iranianum, Pm. scolyti, Pmfraxinopennsylvanicum and Pm. croatiense were isolated and identified. Phaeoacremonium minimum was the most common species and isolated from Gleditsia caspica, Parrotia persica, Cydonia oblonga and Alnus glutinosa. Pathogenicity on selected plants using a mycelium plug inoculation method revealed that most of these species are pathogenic to all the tested trees, with Pm. parasiticum and Pm. minimum being the most pathogenic species. According to host plant species and geographical distribution, the majority of Phaeoacremonium species found represent new records. This is also the first report of Pm. croatiense in Iran and outside Croatia.  相似文献   

6.
Real‐time PCR assays based on the TaqMan system and using ITS sequences were developed for the identification of Phytophthora species, including P. cactorum, P. megasperma, P. plurivora, P. pseudosyringae and P. quercina, all of which are currently causing significant damage to roots of forest trees in both managed stands and natural ecosystems. Total genomic DNA was extracted from mycelia of aforementioned Phytophthora isolates. Species‐specific primers for P. cactorum, P. megasperma, P. plurivora, P. pseudosyringae and P. quercina were designed based on ITS sequences of rDNA. The amplification efficiency of target DNA varied from 93.1% (P. pseudosyringae) to 106.8% (P. quercina). The limit of the detection was calculated as 100 – 1,000 fg DNA, depending on the Phytophthora species. In mixed soil samples, all Phytophthora species were detected for Ct values shifted by 0.7 – 2.1 cycles. Based on these real‐time PCR assays we were able to identify the five Phytophthora species. These techniques will be of value in the identification of these pathogens, which may cause up to 80 – 90% fine root loss in oak stands.  相似文献   

7.
Mass mortality of Fagacean tree species caused by Raffaelea quercivora has occurred widely in Japan. Because conidia or other propagules of the pathogen have not been found in infected trees, pathogen spread is assumed to occur primarily by hyphae. To clarify the relationship between hyphal growth of the pathogen within trees and their vessel arrangements, we examined two native Japanese oaks, Quercus crispula and Quercus glauca, and three exotic American oaks, Quercus coccinea, Quercus palustris and Quercus rubra. Quercus glauca is a radial‐porous species, whereas the other four species have a ring‐porous wood structure. Hyphal growth within inoculated potted living seedlings and in cut, sterilized stem segments of these species was examined microscopically after fungal inoculation. Water conductance in the seedlings was examined using transverse stem sections. The proportion of non‐conductive sapwood in Q. crispula, Q. coccinea and Q. palustris differed between inoculation and control treatment, being much higher in inoculated seedlings. The proportions were positively correlated with the extent of the hyphal growth. In sterilized stem segments, the extent of fungal colonization varied among the foreign ring‐porous species Q. coccinea, Q. palustris and Q. rubra. It is hypothesized that the extent of colonization by R. quercivora reflects the extent of non‐conductive sapwood irrespective of tree species, but is little affected by vessel arrangements.  相似文献   

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Two‐year‐old seedlings of Pinus brutia, P. brutia var. eldarica, Pinus pinea and 3‐year‐old seedlings of Pinus radiata, Pinus sylvestris, Pinus nigra and Cedrus libani were inoculated on the lower stem with isolates of Heterobasidion annosum s.s. collected from the Black Sea and Mediterranean regions of Turkey. In total, 315 seedlings were inoculated in April 2014 and incubated in a growth chamber for 7 weeks at 18–20°C. All isolates were pathogenic on the seven different hosts and had the ability to grow in living sapwood. The isolates had a greater growth on C. libani, P. sylvestris and P. radiata seedlings compared to plants of the other species tested. The least affected species were P. brutia and P. nigra. The isolates originating from the Black Sea region caused longer lesions on the hosts. Overall mortality during 7 weeks of incubation was 4%.  相似文献   

10.
Extracellular proteins were extracted from Phellinus weirii infected Douglas‐fir (Pseudotsuga menziesii var. menziesii) roots and needles to examine endochitinase activity. Chitinases have been associated with the plant's defence response against fungal attack because they hydrolyse chitin, a structural component of fungal cell walls. Protein separation using sodium dodecyl sulphate‐polyacrylamide gel electrophoresis (SDS‐PAGE) followed by Western immunoblot analysis using a polyclonal antibody specific to an endochitinase‐like protein (ECP) resulted in the detection of up to three polypeptides between 27 and 30 kDa in size. Two‐dimensional gel electrophoresis (2‐D PAGE) followed by Western immunoblot analysis revealed that the apoplastic fluid contained multiple ECP isoforms with isoelectric points (pIs) ranging from 5.3 to 5.8 and molecular masses of 27–30 kDa. Chitinase activity in needle and root tissues was measured spectrophotometrically using a colorimetric assay. A gel overlay technique using glycol chitin as a substrate for endochitinase was applied to confirm that the ECP antibody detected an enzymatically active protein. The apoplastic fluid collected from P. weirii‐infected winter Douglas‐fir needles showed anti‐freeze activity and seasonal analysis of needle tissue showed some evidence of ECP accumulation in winter months. ECP was distributed systemically throughout the tree. Increased levels of endochitinase activity in the region of P. weirii infection supports a physiological role for ECP in the plant defence response.  相似文献   

11.
In addition to Hymenoscyphus fraxineus, two fungi identified as Diaporthe eres aff. and Fusarium sambucinum aff. were also isolated from necrotic bark lesions on declining one‐year‐old Fraxinus excelsior in a forest stand in Montenegro. To examine their involvement in ash decline, a pathogenicity test was performed using under bark inoculations on one‐year‐old Fraxinus excelsior. Hymenoscyphus fraxineus was included as comparison. All three fungal species proved highly pathogenic towards one‐year‐old seedlings although lesion sizes differed significantly between the different species. Hymenoscyphus fraxineus was most aggressive, followed by F. sambucinum aff., while D. eres aff. caused the smallest lesions. This study demonstrates for the first time the ability of isolates in the D. eres and F. sambucinum species complexes to cause decline on one‐year‐old common ash seedlings.  相似文献   

12.
A methodological molecular procedure, which included extraction and cloning of the ITS1/2 rDNA of root‐associated organisms with subsequent transformation and sequencing of representative clones, was effective for detection, discrimination and determination of the frequency of the main damping‐off pathogens in roots of Pinus sylvestris seedlings growing in different forest‐tree nursery soils and exhibiting different rates of disease progress. Roots exhibiting slower damping‐off progression were colonized by Fusarium oxysporum, Neonectria radicicola (Ascomycota) and Pythium spp. (Oomycota), which comprised 50% of the microbial community. Roots exhibiting faster damping‐off progression were dominated by Thanatephorus cucumeris (Basidiomycota), which comprised 80% of the microbial community. The microbial community was more diverse in roots with slower damping‐off progression (14 species) than in roots with faster disease progression (seven species).  相似文献   

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The aim of this study was to assess genetic diversity and spatial distribution of 123 strains of the root‐rot pathogen Neonectria macrodidyma isolated from fine living roots of bare‐root nursery cultivated Pinus sylvestris. We found that ITS rDNA sequences were 100% identical for all strains. Arbitrary primed PCR fragment analysis showed the presence of two distinct N. macrodidyma types (Type 1 and Type 2), which included respectively 11 and 14 genotypes composed of 1–29 strains. Results of this study indicate that N. macrodidyma is commonly associated with fine living roots of pine seedlings, is largely disseminated by asexual means of local genotypes and has even distribution in the forest nursery soils. In living roots, N. macrodidyma is likely present as dormant propagules but under favourable conditions, it may develop rapidly and have a significant negative effect on plant health and productivity.  相似文献   

15.
Seed and root rot of pine and cypress seedlings cause heavy annual losses to forest nurseries in Fars Province. Root and crown samples of various species of conifers, such as Tehran pine (Pinus eldarica), brutia pine (Pinus brutia), Arizona cypress (Cupressus arizonica), Shirazian cypress (Cupressus sempervirens var. fastigiata), common cypress (C. sempervirens var. horizontalis) and oriental arborvitae (Thuja orientalis), showing chlorosis, necrosis, stunted growth, defoliation and root and crown rot symptoms were collected from pine and cypress nurseries across Fars province at various time intervals. Infected tissues were washed and cultured on acidified potato dextrose agar (PDA) and corn meal agar (CMA) amended with Delvocide and ampicillin, with and without surface sterilization, respectively, and incubated at 25°C for 3–5 days. Decaying seeds of all plant species were also collected from nursery seed stocks and cultured on PDA. Hyphal tip isolates were used for further studies. Species of Phytophthora, Pythium, Rhizoctonia and Fusarium were isolated from symptomatic seedlings of pine and cypress at different times during the growing season. Pathogenicity of isolates of Phytophthora, Pythium and Rhizoctonia was confirmed on seedlings of all plant species, whereas Fusarium sp. isolated from the seeds of Tehran pine was only pathogenic on seedlings of Tehran pine and Arizona cypress. Virulent isolates identified as Phytophthora nicotiana var. parasitica, Pythium ultimum, Pythium paroecandrum, Rhizoctonia solani and binucleate Rhizoctonia‐like fungus caused root and crown rot of pine and cypress seedlings. Pathogenic isolates of Fusarium identified as Fusarium proliferatum also caused pre‐emergence seed decay of pine and cypress. Isolates of Phytophthora and Pythium were the most virulent of these fungal isolates.  相似文献   

16.
Pinus ponderosa seedlots from eight seed orchards in Western Patagonia, Argentina, were evaluated for the presence of Fusarium species, and isolates obtained were subjected to pathogenicity tests. The following species were isolated: Fusariumoxysporum, Fusarium equiseti, Fusarium verticillioides, Fusarium incarnatum, Fusarium acuminatum, Fusarium graminearum and Fusarium proliferatum. With the exception of F. proliferatum and F. graminearum all Fusarium species were recovered from P. ponderosa seedlings with symptoms of damping‐off. Five Fusarium species were re‐isolated from necrotic roots of seedlings showing chlorosis (F. oxysporum, F. equiseti, F. incarnatum, F. acuminatum and F. verticillioides). High damping‐off losses occurred in Fusarium‐contaminated seed of P. ponderosa and low disease incidence occurred on less contaminated seed. The role of seed‐borne inoculum in disease spread within forest nurseries is discussed.  相似文献   

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The influence of temperature on germination of Quercus ilex acorns in Phytophthora infested soils was quantified for the first time. Radicle damage and mortality of Q. ilex seeds germinating at 17, 20, 23 and 26°C in Phytophthora cinnamomi, P. gonapodyides, P. quercina and P. psychrophila infested soils were assessed and related to in vitro mycelium growth of the same isolates of the pathogens. The optimum growth temperatures of isolates of P. cinnamomi, P. gonapodyides, P. quercina and P. psychrophila were 20–23, 23–26, 20–23 and 20°C, respectively. At 17 and 20°C, all four Phytophthora species caused 100% acorn mortality, whereas at 26°C, acorn mortality was 100, 10, 25 and 0% in P. cinnamomi, P. gonapodyides, P. quercina and P. psychrophila infested soils, respectively. At 23°C, P. cinnamomi and P. gonapodyides reduced acorn radicle length more than P. quercina and P. psychrophila, whereas at 26°C, only P. cinnamomi caused further reduction in radicle length. The higher susceptibility of germinating acorns in comparison to seedlings reported in the literature indicates age‐related susceptibility of Q. ilex to Phytophthora. The seedling/pathogen growth ratio was inversely related to the reduction in radicle length at different temperatures ( = 0.84, p < 0.0001), suggesting that rapid germination may allow seedlings to escape from infection. Increasing temperatures had different effects on damage to acorns depending on the pathogen present in the soil, indicating that Phytophthora species × temperature interactions determined Q. ilex germination. The effects of temperature on the impacts of Phytophthora species based on climate change predictions for Mediterranean countries are discussed.  相似文献   

19.
The ecological importance of riparian forests is well known. However, these forest habitats have been disturbed by human activities over the past century as a result of the introduction of flow regulations. Mortality of the riparian alder population caused by Phytophthora has become an important issue in Europe in the past two decades. The main objective of this study was to examine the pathogenicity of the Phytophthora alni complex (P. xalni, P. uniformis and P. xmultiformis) and P. plurivora in Alnus glutinosa seedlings. Phytophthora alni complex has traditionally been identified as the main causal agent of alder decline; however, in this study, P. plurivora was found to cause as much damage in inoculated seedlings as P. alni complex. In fact, Spanish isolates of P. xalni caused mortality rates of ca. 30%, whereas P. plurivora killed ca. 50% of seedlings. Likewise, there were no differences between P. xalni ‐ and P. plurivora‐inoculated seedlings in either the length of lesion or biomass allocation. The pathogenicity of the species within P. alni complex did not differ significantly. The findings confirm that P. plurivora, which is one of the most common species of Phytophthora found in European nurseries, is highly pathogenic to alder seedlings. Urgent implementation of measures to prevent the risk of spread of the pathogen from nurseries to natural ecosystems is required. Further studies are required to clarify the role that P. plurivora is playing in alder decline both individually and in combination with P. alni complex.  相似文献   

20.
Ophiostomatoid fungi are carried by various bark beetles. However, very little is known about the role of these fungi in conifer roots. We studied ophiostomatoid fungi in roots of dying and dead Pinus sylvestris trees and tested the potential phytotoxicity of some isolates using a sensitive bioassay with Lepidium sativum in Poland. Fungi were identified based on their morphology and DNA sequencing. Three ophiostomatoid fungi, Leptographium procerum, Sporothrix inflata and Ophiostoma pallidulum, were isolated from the roots. The most abundant soil‐borne fungus, S. inflata, and relatively rare O. pallidulum were isolated for the first time from roots of dying and dead pine trees. The frequency of S. inflata and O. pallidulum correlated with tree decline. The fungi were isolated more frequently from roots of dead than dying trees. Sporothrix inflata and O. pallidulum slightly reduced the stem and root growth of L. sativum. Leptographium procerum reduced more significantly root than stem growth. This species reduced root elongation 32–54% after 10–17 days of incubation.  相似文献   

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