Brenneria spp. and Rahnella victoriana associated with acute oak decline symptoms on oak and hornbeam in Iran

Decline diseases of forest trees are complex syndromes not attributable to single causal factors. In Iran, symptoms of decline disease have been observed in a number of native forest species including Quercus castaneifolia (chestnut‐leaved oak), Q. brantii (Persian oak) and Carpinus betulus (hornbeam). The symptoms are prevalent in the northern forests and the Zagros mountain forests. There are parallels between the disease in Iran and acute oak decline (AOD) reported in the UK, specifically the presence of weeping cankers, which have been attributed to a polybacterial complex wherein Brenneria goodwinii is considered a key necrogen. Based on the AOD symptomatology, and as a first step towards discovering potential causal agents of the stem weeping symptoms of affected trees in Iran, necrotic tissues were tested primarily for the presence of B. goodwinii. Symptomatic Q. castaneifolia and C. betulus from the Mazandaran Province and symptomatic Q. brantii from Kohgiluyeh and Boyerahmad Province were sampled. Isolation and culture on a selective medium yielded uniform bacterial colonies. Isolates were characterized using phenotypic and genotypic (DNA sequencing) tests. The isolates were phenotypically identical to members of Pectobacteriaceae and Yersiniaceae, specifically Brenneria and Rahnella spp. The nucleotide sequences of the 16S rRNA and housekeeping genes gyrB, infB and atpD (MLSA) amplified via PCR demonstrated that the isolates from the trees in Iran were indeed B. goodwinii, B. roseae subsp. roseae, Rahnella victoriana and an unknown species of Brenneria. Most bacteria isolated from non‐symptomatic trees were Gram‐positive, and Pseudomonas spp. were dominant, but bacterial species isolated from the diseased trees were not detected in healthy trees. Hypersensitivity response tests were positive, but inoculation on saplings was more variable with internal necrosis developing only once in the test period. Therefore, further testing is required. This is the first report of the incidence of B. goodwinii, B. roseae subsp. roseae, R. victoriana and Brenneria sp. associated with acute oak decline‐like symptoms on Q. castaneifolia, Q. brantii and C. betulus across the western forests of Iran and in the world.     

Bacillus pumilus and Stenotrophomonas maltophilia as two potentially causative agents involved in Persian oak decline in Zagros forests (Iran)

The oak decline is known as one of the most destructive complex diseases causing high economic losses around the world, especially in Iran. The main objective of the present study was to investigate the possible role of bacteria as causative agents of oak decline in the Zagros forests of Iran. To do this, stem, root and leaf samples were taken from symptomatic Persian oak trees (Quercus brantii) in different zones of Zagros forests (Ilam Province, Iran). From 150 bacterial isolates, 20 showed pathogenicity against Geranium seedlings. Among 20 hypersensitivity test positive strains, four strains showed pathogenicity against oak saplings. Based on morphological and 16S rRNA gene sequence analysis, three strains were identified as Bacillus pumilus and one strain as non‐sporulating Gram‐negative Stenotrophomonas maltophilia. Pathogenicity studies of different B. pumilus and S. maltophilia strains revealed that they have potential to cause the disease in oak saplings and symptoms of disorder in Persian oak trees. To our knowledge, there are no previous records of B. pumilus and S. maltophilia causing decline on Fagaceous trees like Q. brantii. More detailed field and molecular studies are required to confirm the absolute role of such bacteria in occurrence of oak decline in Zagros forests.     

Molecular and morphological characterization of Ektaphelenchoides maafiae n. sp. (Nematoda: Ektaphelenchinae) from northern forests of Iran

Ektaphelenchoides maafiae n. sp. was isolated during a survey of nematodes associated with bark samples of an oak tree (Quercus castaneifolia) in Gorgan, Golestan Province, northern Iran. The new species has a body length of 480–546 μm (in females) and 431–480 μm (in males). The cuticle is weakly annulated, with three lateral lines. Lip region offset. The stylet is 13–15 μm long without basal swellings. The excretory pore is located at the level of the metacorpus base to slightly posterior, and hemizonid is at 15–17 μm posterior to the excretory pore. The post‐uterine sac is short, 6–8 μm long. Spicules having rounded condylus, rostrum short, conical with bluntly pointed tip, a cucullus (apophysis) presented on the dorsal distal end. Male tail bearing four (2 + 2) caudal papillae, conical, with sharply pointed terminus. The new species is close to four known species of the genus, including E. hunti, E. ruehmi, E. caspiensis and E. poinari, but differs from them by body size, shape of tail terminus, stylet length, shape and size of spicules, length of post‐vulval uterine sac and number of caudal papillae. Phylogenetic analysis based on small subunit (SSU) and partial large subunit (LSU) sequences of rDNA confirmed its status as a new species.     

First detection of Neoscytalidium dimidiatum associated with canker disease in Egyptian Ficus trees

In September 2013, a canker disease of Ficus nitida and F. benjamina was reported for the first time in Assiut governorate, Egypt. Infected samples were collected from various locations. Pure cultures of a fungus were isolated on potato dextrose agar at 25°C from diseased plants. Morphological investigation and DNA sequencing showed that the causal agent was Neoscytalidium dimidiatum. A pathogenicity test conducted using 2‐year‐old plants of each host gave 70%–80% infection, and the pathogen was reisolated from the inoculated plants. A pure culture of N. dimidiatum was deposited in the culture collection of the Assiut University Mycological Centre (AUMC) under the code AUMC 9293 and the ITS sequence was placed in NCBI under accession number KX985929 .     

Characterization of endophytic bacteria with plant growth promotion and biological control potential isolated from walnut trees

In this study, a total of 68 endophytic bacteria were isolated from different tissues of walnut trees. About 55% and 22% of the strains had the ability to produce indole acetic acid and gibberellic acid, respectively. The capability of isolates to solubilize phosphate, growth on N‐free medium, siderophore, protease and lipase production was varied. Based on phenotypic grouping and plant growth promotion properties, twelve isolates were selected and 16S rRNA gene‐based phylogenetic analysis revealed that strains showed 99%–100% similarity to Pseudomonas, Bacillus, Arthrobacter, Roseomonas and Streptomyces genera. Amongst the selected strains, PS12, KS54, JS66 and KS74 showed root and shoot growth enhancement of poplar cutting. NS70, KS54 and PL36 strains showed antagonistic activity against Pseudomonas syringae pv. syringae;RR47, KS74 and NR69 strains had inhibition effects against Brenneria nigrifluens; and JS66 and RR26 strains had antagonistic activity against both phytopathogens under in vitro conditions. This is the first reported study to elucidate the endophytic bacterial diversity associated with walnut trees with beneficial attributes.     

Characterization,pathogenicity and host range studies of Paecilomyces formosus associated with dieback of Christ's thorn trees (Paliurus spina-christi Mill.) in Iran

Christ's thorn (Paliurus spina-christi Mill.) tree is an important and valuable forestry species and has significant potential in afforestation and reforestation systems. In recent years, dieback symptoms were observed on Paliurus spina-christi trees in Gilan-e Gharb region of Kermanshah province, west Iran. During 2020 to 2021, Christ's thorn trees that exhibit branch dieback symptoms were sampled for fungal isolations. The pathogenicity tests, biochemical assay of growth, acid production on creatine sucrose agar (CREA) medium and morphological and molecular investigations (ITS and β-tubulin regions) identified Paecilomyces formosus as a dieback causal agent. This is the first report of P. formosus pathogenicity on Paliurus spina-christi trees. Furthermore, disease severity was assayed on 11 tree species under three different temperatures 15, 25 and 35°C. Disease severity caused by all isolates on detached branches was greater at 35°C than at 15 and 25°C. This study presents the host range of this pathogen and showed that these potential hosts are prone to this pathogen under high temperatures, which forest trees experienced in recent decades.     

“Candidatus Phytoplasma solani” associated with Eucalyptus witches’ broom in Iran

During summer of 2015, Eucalyptus camaldulensis plants showing witches’ broom, little leaf and general yellowing of the foliage were observed in west of Fars and Khozestan province of Iran. DNA from samples of 22 symptomatic and two asymptomatic trees was extracted and subjected to molecular analyses. Nested‐PCR test using R16F2n/R16R2 primers confirmed phytoplasma presence in 63% of symptomatic Eucalyptus plants. Sequence analysis along with virtual RFLP of the 16S ribosomal DNA allowed to classify three Eucalyptus witches’ broom strains into the “stolbur” (“Candidatus phytoplasma solani”) 16SrXII‐A subgroup. Comparison of the secA and secY gene sequences with sequences deposited in GenBank confirmed the phytoplasma identity. Real and virtual RFLPs of the amplified secY gene using HaeIII, MseI and RsaI restriction enzymes showed profiles indistinguishable from each other. This is the first study reporting E. camaldulensis as a new host species for “Ca. P. solani.”     

Host specificity of co‐infecting Botryosphaeriaceae on ornamental and forest trees in the Western Balkans

The Botryosphaeriaceae is a diverse family of endophytes and fungal pathogens of mainly woody plants. We considered the host range and distribution of these fungi by sampling diseased ornamental and forest trees and shrubs in Serbia, Montenegro, Bosnia and Herzegovina, spanning a Mediterranean and a Continental climatic region. In total, ten Botryosphaeriaceae species were identified in the Western Balkans and with the exception of Sphaeropsis visci and Phaeobotryon cupressi, which occurred on one host, all the species had a broader host range. Phaeobotryon cupressi was found only in the Mediterranean region and S. visci, Dothiorella sp., Dothiorella sarmentorum and Diplodia seriata were present only in the Continental region. Pathogenicity tests were conducted on a variety of hosts from which the Botryosphaeriaceae species were isolated. These included leaves and/or stems of seedlings of 21 hosts, and cut leaves and/or branches of six hosts. Moreover, stems of seedlings of Chamaecyparis lawsoniana, Cedrus deodara, Picea omorika, Pinus patula and Eucalyptus grandis were inoculated as hosts from which some or all of the Botryosphaeriaceae species used for inoculation were not isolated. Inoculations showed that the majority of these fungi could also co‐infect hosts other than those from which they were isolated. The results suggest that most of the species have broad host ranges and can potentially cause disease on a broad range of tree species under certain conditions.     

The true nature of Ganoderma in Iran: Taxonomy based on ITS and mtSSU rDNA

The genus Ganoderma Karst. has broad‐spectrum usage in biotechnology, medicine and the food industry. The complexity of the morphology within species has led to uncertain identification in the past, but recent advancements in molecular identification methods have provided scientists with the opportunity to better understand the taxonomy of the species. The present study attempts for the first time to elucidate the distinctiveness of the Ganoderma species growing in Iran concerning those elsewhere in the world based on mitochondrial small subunit ribosomal DNA (mtSSU rDNA) and internal transcribed spacer (ITS) sequence data. The results disclosed that the G. lucidum Karst. samples collected in Iran are more similar to the European Ganoderma species than to the Asian (Chinese) one used in this study.     

An integrative study of Sakia sisanganensis n. sp. (Rhabditida: Tylenchidae) from Sisangan forest,Iran, and new morphological observations for the genus

Further data on the morphology (the lip region characters) and phylogeny of the genus Sakia are presented. The new observations were based on scanning electron microscopy (SEM) and light microscopy (LM). A new species, Sakia sisanganensis n. sp., was recovered from rotten wood of a dead beech tree (Fagus orientalis) in northern Iran, herein described and illustrated based on an integrative approach, that is morphological, morphometric and molecular characters. The new species is characterized by a combination of the following features: fine transverse striae and vestigial single band in the lateral field in SEM. Labial area dorso‐ventrally flattened. Oral region with two concentric hexagonal plates, the inner one apparently elevated. Amphidial openings short, slit‐like. Stylet delicate. Median bulb fusiform to spindle‐shaped with weak valvular apparatus. Spermatheca functional. Tail filiform with faintly pointed tip and males common. The new species was morphologically compared with four known species of the genus, viz., S. alii, S. arboris, S. castori and S. indica, all having indistinct lateral fields. Molecular phylogenetic analyses were performed based on the small subunit ribosomal RNA gene (SSU rDNA). In the Bayesian inference (BI), S. sisanganensis n. sp. with two isolates was strongly supported as a sister taxon of a clade harbouring S. arboris + Lelenchus species. However, in the maximum likelihood (ML) analysis, the new species formed a clade with S. arboris, thus supporting the reciprocal monophyly of the genera Sakia and Lelenchus. Accordingly, the test of monophyly was performed (using Bayes factor) and the results did not reject the monophyly of sakia (i.e., S. sisanganensis n. sp. and S. arboris as sister taxa) based on the currently available data.     

Wood‐rotting basidiomycetes are a minor component of fungal communities associated with Acacia hybrid trees grown for sawlogs in South Vietnam

Acacia hybrid (Acacia mangium × Acacia auriculiformis) clones are widely planted in Vietnam with a total of approximately 400,000 ha to meet the demand for pulpwood, sawn timber and wood chip exports. Silvicultural techniques such as pruning and thinning have been applied to improve productivity and sawlog quality of Acacia hybrid plantations. However, those techniques may also create opportunities for wood decay fungi to enter the Acacia hybrid stems through wounds and cause stem defects that reduce sawlog quality and the value of the plantation. The presence of fungal decay agents in Acacia hybrid trees was examined in two Vietnamese plantations. In July 2011, just prior to a second thinning, discoloured wood samples were taken from a three‐year‐old Acacia hybrid plantation at Phan Truong Hai for the isolation of fungi. In July 2012, approximately 18 months after pruning and thinning treatments, discoloured wood samples were taken from a three‐year‐old Acacia hybrid plantation at Nghia Trung for the isolation of fungi. DNA sequencing of the rDNA ITS identified the isolates. In May 2015, approximately 4 years after thinning and fertilizer treatments, discoloured and decayed wood samples were taken from the above (7‐year‐old) Acacia hybrid plantation at Phan Truong Hai for fungal identification. DNA was extracted directly from discoloured and decayed wood samples and fungal rDNA ITS amplicons sequenced on a Roche 454 sequencer. The results showed that silvicultural treatments did not affect the fungal communities associated with discoloured and decayed wood of Acacia hybrid plantation at Phan Truong Hai. A total of 135 fungal species or OTUs (operational taxonomic units) were identified, including 82 members of Ascomycota and 52 Basidiomycota.     

First record of Ektaphelenchoides pini associated with Ips sexdentatus on Pinus nigra laricio in Italy

In February 2015, an unexpected windstorm downed five hectares of a European black pine Pinus nigra subsp. laricio forest formation located close to Vallombrosa, Florence (Central Italy). In the following spring, an extensive survey was conducted in the area. Felled trees, stumps and all the suitable plant material were screened for the presence of the pinewood nematode (PWN), Bursaphelenchus xylophilus, by sampling wood and bark. Bark beetles were then collected from the gallery systems on the inner side of bark samples and observed in the laboratory. The following bark beetles were morphologically identified: Ips sexdentatus, Orthotomicus erosus, O. laricis and Pityogenes bidentatusa. The dissection of Ips sexdentatus allowed the extraction of numerous nematodes that were morphologically and molecularly identified as Ektaphelenchoides pini. Conversely, only few nematode specimens were isolated from either pine bark or wood. These individuals could be only molecularly identified and belonged to an undescribed nematode taxon. Even though no PWN was recorded in the investigated sites, our survey allowed the detection of a new association between E. pini and I. sexdentatus on P. nigra.     

Chestnut Red Stain: Identification of the fungi associated with the costly discolouration of Castanea sativa

In Europe, fungal pathogens have reduced the overall productivity of sweet chestnut (Castanea sativa) stands and continue to threaten the economic viability of forestry operations. Chestnut Red Stain (CRS) in north‐eastern Spain, locally referred to as Roig, is capable of decreasing the market value of chestnut timber to the point of rendering chestnut coppices uneconomical. Despite its economic importance, the specific cause of this red discolouration is unknown. With the objective of verifying the presence of fungi within the symptomatic wood, and identifying the fungus or suite of fungi associated with the red stain, wood samples were collected and cultured from 37 stumps found in eight recently harvested stands in the Montseny and Montnegre‐Corredor Natural Parks. To separate the fungi associated with CRS from other species inhabiting the chestnut wood, the origin of each fungal culture was mapped in every stump. The fungi were isolated from cultures and identified by sequencing the ITS region. The results provide insight into the fungal community inhabiting chestnut wood and the potential cause of CRS; nine species were identified including two species known to cause decay in chestnut. One of them, Fistulina hepatica, appears to be a likely candidate for the causal agent of CRS. This is the first study reporting the fungi associated with CRS and opens the door to new epidemiological studies focused on F. hepatica.     

Fungi associated with Cyclaneusma needle cast in Scots pine in the west of Poland

Needles of Pinus sylvestris with and without symptoms of Cyclaneusma needle cast, from the west of Poland, were examined for abundance and diversity of fungi using Illumina sequencing. Fungal communities were dominated by Ascomycota (93.6%–98.6% of OTUs). Basidiomycota and non‐culturable fungi were less frequent. Needles were colonized by 260 taxa. Ascomycota, Basidiomycota and Glomeromycota were represented by 149, 39 and 1 taxa. Abundance of fungi was least in 1.5‐year‐old needles attached to twigs and greatest in fallen 2‐year‐old needles. Fungal communities had least diversity in 1.5‐year‐old needles and most diversity in 0.5‐year‐old needles of current growth. It was found that (a) the most common fungi were the needle pathogens Cyclaneusma minus, Lophodermium spp. and Sydowia polyspora; (b) less common potential pathogens were Cenangium ferruginosum, Coniothyrium complex, Desmazierella acicola, Neocatenulostroma germanicum and species in the genera Neodidymelliopsis, Pestalotiopsis, Phoma, Pleurophoma and Pyrenochaeta; (c) common primary or secondary saprotrophs included species of Alternaria, Aureobasidium, Beauveria, Cladophialophora, Cladosporium, Epicoccum, Exophiala, Lecanicillium, Penicillium, Cryptococcus and Kwoniella; (d) Lophodermium was represented mostly by Lophodermium pinastri which occurred 4–72 times more frequently than Lophodermium seditiosum; (e) frequencies of C. minus and C. ferruginosum were lower in the 0.5‐year‐old symptomless needles, increased in the symptomatic and symptomless 1.5‐year‐old needles and decreased after needle fall; (f) frequency of L. seditiosum was highest in 0.5‐year‐old needles; (g) frequency of L. pinastri increased with needle age whereas S. polyspora increased after needle fall; (h) lower frequency of L. pinastrii was associated with higher frequency of S. polyspora. It was concluded that Cyclaneusa needle cast in Poland may be caused by C. minus accompanied by C. ferruginosum, L. seditiosum, L. pinastrii and S. polyspora. Participation of Coniothyrium spp., Epicoccum nigrum, Pestalotiopsis spp. and Phoma spp. in the disease progress cannot be excluded.     

Identification and characterization of fungi associated with internal wood lesions and decline disease of willow and poplar trees in Iran

During the study of fungal trunk pathogens associated with urban trees decline in Shiraz (Iran), a serious decline of willow and poplar trees was observed. Therefore, an investigation was conducted on these trees in some areas of this city during spring and summer 2012 and 2013, to determine the main fungal trunk pathogens associated with these ornamental plants. Plant materials were collected from trees exhibiting disease symptoms such as yellowing, shoot canker, shoot dieback, defoliation and internal wood necrosis and decayed wood. Fungal isolations were made from discoloured or decayed wood tissue onto 2% malt extract agar (MEA) amended with streptomycin sulphate. Nine species, Fomes fomentarius, Diplodia seriata, Lasiodiplodia theobromae, Dothiorella sarmentorum, Neoscytalidium hyalinum, Diatrype whitmanensis, Phaeoacremonium rubrigenum, P. aleophilum and P. parasiticum, were identified based on morphology and DNA sequence comparisons. Pathogenicity tests were performed on detached shoots of willow and poplar trees under greenhouse conditions. Lasiodiplodia theobromae caused the longest lesions on willow. On poplar shoots, the longest lesions were caused by P. parasiticum. Diplodia seriata produced the smallest lesions on both woody hosts. First reports from willow wood include P. parasiticum, P. rubrigenum, D. whitmanensis, L. theobromae, D. seriata and N. hyalinum, while new reports from poplar wood include P. parasiticum and Do. sarmentorum. Based on our knowledge, this is also the first report of D. whitmanensis in Iran.     

暖温带北缘3种栎树径向生长对气候要素的响应差异及预测

目的 比较同一气候区不同栎类树种生长对气候要素的响应差异,特别是在分布边缘不同树种的种内种间差别。 方法 利用树木年轮学方法,分析了我国暖温带北缘3种优势落叶栎树（蒙古栎、栓皮栎和麻栎）径向生长对气候要素的响应差异;同时利用混合线性效应模型构建了3个树种径向生长与气候的关系,并基于未来气候情景（RCP2.6和RCP8.5）预测了其未来生长趋势。 结果 同一地区栎类树种年表具有较高的相似性,而不同地区间差异较大;在径向生长对气候响应方面,同一地区的不同栎类树种和不同地区同一栎类树种也存在一定差别。其中北京地区的3种栎类树种的径向生长与当年6月气温呈负相关关系,但在辽宁仙人洞地区却与气温无显著相关关系;北京地区的麻栎、河北地区的蒙古栎、辽宁大黑山地区的栓皮栎的径向生长均受降水条件限制。模型预测显示,至本世纪末,北京地区和辽宁大黑山地区栎类树种的径向生长呈现下降趋势;而辽宁仙人洞地区栎类树种径向生长呈增加趋势。 结论 我国暖温带北缘不同地区的栎类树种对气候要素的响应存在显著差别,主要为取样地区的气候所支配,下一步尚需结合林分及立地因子,来厘清各类环境要素的相对贡献度。     

Foliar nematode,Litylenchus crenatae ssp. mccannii,population dynamics in leaves and buds of beech leaf disease‐affected trees in Canada and the US

A foliar nematode, Litylenchus crenatae ssp. mccannii, is associated with beech leaf disease (BLD) symptoms. Information about the types of tissues parasitized and how nematode populations fluctuate in these tissues over time is needed to improve surveys as well as understand the nematodes role in BLD. During this study, the nematode was detected throughout the known range of BLD by researchers at both Canadian and US institutions using a modified pan method to extract nematodes. Monthly collections of symptomatic and asymptomatic leaves during the growing season (May–October), and leaves and buds between growing seasons (November–March), revealed that nematodes were present in all tissue types. Progressively larger numbers of nematodes were detected in symptomatic leaves from Ohio and Ontario, with the greatest detections at the end of the growing season. Smaller numbers of nematodes were detected in asymptomatic leaves from BLD‐infected trees, typically at the end of the growing season. The nematode was detected overwintering in buds and detached leaves. The discovery of small numbers of nematodes in detached leaves at one location before BLD was detected indicates that nematodes may have been present before disease symptoms were expressed. Other nematodes, Plectus and Aphelenchoides spp., were infrequently detected in small numbers. Our findings support the involvement of the nematode in BLD and indicate that symptoms develop only when certain requirements, such as infection of buds, are met. We also found that the nematode can be reliably detected in buds and leaves using the modified pan extraction method.     

Phytophthora species associated with dieback of sweet chestnut in Western Turkey

Sweet chestnut (Castanea sativa) is an important tree species in the Marmara and Aegean regions of Turkey as these two regions produce the great majority of edible nuts, especially those used for marron glacé production. Chestnut forests and orchards in these regions showing severe dieback symptoms not associated with chestnut blight were investigated to determine the role of Phytophthora spp. in the decline syndrome. Soil samples were collected from around 108 symptomatic chestnut trees at 29 sites and Phytophthora spp. isolated using soil baiting technique and selective medium. Species isolated were identified by cultural characteristics and ITS sequencing. Phytophthora x cambivora was the dominant species detected in 13 sites, followed by P. cinnamomi (5 sites), P. plurivora (3 sites) and P. cryptogea (1 site). Phytophthora x cambivora was present in both regions, while P. cinnamomi was found only in the Marmara region in coastal areas around Istanbul. When inoculated at the stem bases of 3‐year‐old chestnut saplings, P. cinnamomi produced significantly longer necrotic lesions (7.8–12.0 cm) than P. x cambivora (2.6–6.3 cm) by 12 days after inoculation. Phytophthora plurivora was the least aggressive species causing only small lesions. Phytophthora cryptogea, which represents the first record on chestnut in Turkey, produced intermediate sized lesions in between P. x cambivora and P. plurivora. These results indicate that P. x cambivora and in some areas P. cinnamomi play major roles in the observed dieback of sweet chestnut in western Turkey.     

Occurrence and diversity of Phytophthora species in declining broadleaf forests in western Ukraine

In western Ukraine, forest decline and dieback of several broadleaved tree species have become increasingly evident during recent years, and surveys in some areas have shown symptoms indicative of Phytophthora infections. In this study, we aimed to determine the occurrence and diversity of Phytophthora species associated with several broadleaved tree species (Alnus glutinosa, Betula pendula, Castanea sativa, Fagus sylvatica and Quercus robur) from forest stands where dieback has been observed. Rhizosphere soil samples were collected from 14 forest stands during 2017 and 2018 and tested for the presence of Phytophthora species using morphological and molecular methods. Seven Phytophthora species (P. bilorbang, P. cactorum, P. gallica, P. gonapodyides, P. lacustris, P. plurivora and P. polonica), and two other clade six taxa were detected from the various forest types, several of which are probable agents responsible for decline. Four of the Phytophthora species (P. bilorbang, P. gallica, P. plurivora and P. polonica) have previously never been reported from broadleaf forests in Ukraine.