First report of Lecanosticta acicola on non‐native Pinus mugo in southern Sweden

In 2017, severe symptoms of brown spot needle blight, similar to those caused by Lecanosticta acicola, were observed on needles of non‐native Pinus mugo var. Hesse planted in an arboretum in southern Sweden. Microscopic characterization and molecular diagnostics of isolates obtained from diseased needles confirmed the identity of L. acicola. This is the first report of the quarantine pathogen L. acicola in Sweden. Subsequent surveys are urgently needed to assess the presence and extent of the L. acicola invasion in Scots pine (Pinus sylvestris) stands in Sweden and other Scandinavian countries.     

Dothistroma spp. in Western Ukraine and Georgia

Dothistroma needle blight (DNB) is among the most serious foliar diseases affecting Pinus spp. globally. Infected needles were collected from potential host species in four locations in western Ukraine and in four locations in eastern Georgia during spring–summer 2015 to update the knowledge on pathogen distribution in these countries. Dothistroma spp. were detected using isolation, sequencing and species‐specific priming (SSPP) PCR. Two new hosts for Dothistroma spp. were recorded in western Ukraine: D. septosporum on Pinus nigra var. australica and D. pini on P. nigra var. mollet. D. septosporum was found on 15‐year‐old P. strobus in western Ukraine. New hosts for D. septosporum were recorded in Georgia on 5‐ to 10‐year‐old naturally regenerated P. sylvestris var. hamata and on 40‐ to 50‐year‐old P. ponderosa trees. D. pini was found for the first time in Georgia on 30‐ to 40‐year‐old P. nigra trees. The work confirmed the presence of both D. septosporum and D. pini in western Ukraine and Georgia, and demonstrated new hosts for both Dothistroma species.     

Infection approach of Diplodia sapinea,the causal agent of tip blight of Pinus tabulaeformis in China

To clarify the infection approach of Diplodia sapinea, a pathogen that causes tip blight of Pinus tabulaeformis, the infection process of the pathogen in needles was observed using scanning electron microscopy (SEM). In addition, the disease incidence on branches damaged by Aphrophora flavipes (Hemiptera: Cercopidae) and Dioryctria splendidella (Lepidoptera: Pyralidae) in the forest was also investigated. Then, branches and needles of P. tabulaeformis were inoculated using the D. sapinea spore suspension under indoor and field conditions. The results showed that the damage caused by A. flavipes could aggravate the occurrence of tip blight of P. tabulaeformis to some extent. Moreover, the pathogen could also penetrate 1‐, 2‐ and 3‐year‐old pine needles through stomata in the field. The pathogen infected the 1‐year‐old branches first and then gradually spread to 2‐ and 3‐year‐old branches.     

Needle blight of pine caused by two species of Dothistroma in Hungary

Dothistroma septosporum and D. pini cause the serious pine needle disease, Dothistroma needle blight (DNB). Of these, D. septosporum has a global distribution, while D. pini is known only from the USA, Russia, Ukraine and France. During a study of D. septosporum isolates from Pinus nigra in Hungary, microsatellite markers revealed the presence of a second Dothistroma species. The aim of this study was to identify the DNB pathogens occurring in Hungary using four different molecular techniques. These included sequencing of the rDNA ITS region, a species‐specific ITS‐RFLP, mating type primers and a diagnostic microsatellite marker, Doth_A. Results showed that both D. septosporum and D. pini occur in Hungary and that both DNB pathogens were present on the same trees and in some cases, the same needles. Mating types MAT1 and MAT2 of D. septosporum were shown to be present in Hungary, but only the MAT2 of D. pini was found. In addition, examination of needles with DNB symptoms from Russia revealed the presence of D. pini on Pinus pallasiana and P. nigra as well as on a new host, Pinus mugo. The molecular markers applied in this study were sufficiently robust to identify and differentiate between the two DNB pathogens, both in culture and directly from needles. They will consequently be useful to document the geographical range and to monitor the spread of D. septosporum and D. pini in future studies.     

Identification and pathogenicity of Alternaria alternata causing leaf spot and blight disease of Ailanthus excelsa in India

Alternaria leaf spot of Ailanthus excelsa is generally considered as minor disease in India. Recently, severe disease outbreaks were recorded in the nursery of the Forest Research Institute, Dehradun, progeny trial at Jhumpa, Haryana, and in a nearby farm field. Leaf symptoms included small circular, brown, necrotic spots with a chlorotic halo. With severe infections, leaf spots coalesced and resulted in leaf blight. A small‐spored Alternaria with concatenated conidia was isolated consistently from the leaf samples with spot symptoms. Sequence analyses of the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) and the translation elongation factor 1‐alpha (tef‐1α) gene region of two fungal isolates confirmed the species as A. alternata. In detached leaf assays, both isolates produced symptoms similar to those observed on the nursery/field‐grown plants. To validate the detached leaf assay result, pathogenicity was also demonstrated on whole plants in a glasshouse. Koch's postulates were fulfilled by re‐isolating A. alternata from the inoculated leaves. This work is the first to confirm that A. alternata is associated with leaf spot and blight disease of A. excelsa in India.     

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.     

First report of Mycosphaerella dearnessii on Pinus nigra var. nigra in Austria

Mycosphaerella dearnessii, the causal agent of brown‐spot needle blight (BSNB), has been known to occur in Austria since 1996. In 2011, the disease was for the first time confirmed on Austrian pine (Pinus nigra var. nigra) in this country. As previous records are doubtful, this may also be the first definite report of M. dearnessii on P. nigra in entire Europe.     

Occurrence of leaf spot disease caused by Neopestalotiopsis clavispora on Taxus chinensis in China

Taxus chinensis is a valuable tree with uses for timber, medicinal, ornamental and landscape purposes. Most importantly, it is a source of the anti‐cancer drug paclitaxel. In 2016, needle spot disease was observed on T. chinensis in Fujian Province of China. Five isolates of similar morphology were isolated from diseased samples. The pathogen was identified as Neopestalotiopsis clavispora based on morphological and cultural characteristics and confirmed by sequence analysis of the internal transcribed spacer region (ITS) and the large subunit ribosomal RNA gene (LSU). The pathogenicity of the isolates was confirmed by fulfilling Koch's postulates. To the best of our knowledge, this is the first report to indicate that the causal agent of needle spot disease on Chinese yew in China is N. clavispora.     

Modelling of the spread of Dothistroma septosporum in Europe

Dothistroma needle blight (DNB), a disease affecting several pine species, is currently generating great concern in Europe. Caused by Dothistroma pini and Dothistroma septosporum, DNB affects pine needles and causes premature defoliation, which results in growth reduction and, in extreme cases, mortality. The disease has increased in importance in Europe over the last 20 years, with an increase in the number of observations of DNB in regions with large areas of Pinus sylvestris in northern Europe. This article presents a cell‐based spatiotemporal model for predicting the likelihood and intensity of the future spread of D. septosporum in Europe. Here, “spread” includes both invasion of new regions and infection of healthy stands within already‐colonized regions. Predicted spread depends on the availability of host species, climatic suitability of different regions to D. septosporum and dispersal of sexual and asexual spores from infected trees to surrounding forests via water splash, mist and wind. Long‐distance spread through transport of infected seedlings is also included in the model. Simulations of spread until 2007 and 2015 were used to validate the model. These simulations produced similar patterns of spread to those observed in Europe. Simulations for 2030 suggested that additional and new outbreaks are likely to occur in Scotland, southern Norway, southern and central Sweden, northern parts of Germany and Poland, Estonia, Latvia and south‐west Finland. Preventing the delivery of infected seedlings would be an effective method for reducing the spread of D. septosporum in the Nordic countries, Scotland and Ireland, the Baltic countries, and parts of Germany, Poland and Belarus. In these states, prevention of transport of infected seedlings can reduce the probability of additional spread by 15%–40%.     

New hosts for Lecanosticta acicola and Dothistroma septosporum in newly established arboreta in Spain

A historical outbreak of needle blight disease was recorded during 2018 to 2019 in plantations of Pinus radiata and Pinus nigra in the North of Spain. The main pathogens involved in this historical outbreak were identified as Lecanosticta acicola and Dothistroma septosporum. Recently, a variety of tree species in three arboreta planted between 2011 and 2013 in the Basque Country as part of the European project REINFFORCE were showing symptoms of needle blight and defoliation. The aim of this study was to determine which pine species were affected with these pathogens. Tree species sampled included several provenances of P. brutia, P. elliottii, P. nigra, P. pinaster, P. pinea, P. ponderosa, P. sylvestris and P. taeda. Using molecular identification methods, Lecanosticta acicola was confirmed infecting Pinus brutia (Provenance: Alexandropolis, Greece and var. eldarica, Crimea) and represents a new host species for this pathogen. Pinus elliottii (Provenance: Georgia, USA) and P. ponderosa (Provenance: Central California, USA) are new host reports of L. acicola for Spain. Dothistroma septosporum was found for the first time on P. brutia (Provenance: Marmaris, Turkey) and P. ponderosa (Provenance: Oregon, USA) in Spain and was also detected infecting P. nigra (Provenance: Sologne Vayrières, France).     

Mycobiota in needles of Abies alba with and without symptoms of Herpotrichia needle browning

The ascomycete Nematostoma parasiticum (syn.: Herpotrichia parasitica) is commonly perceived as the causal agent of the so‐called Herpotrichia needle browning in silver fir (Abies alba). However, its fruitbodies are rarely present on symptomatic needles, which are also colonized by many presumably saprotrophic fungi. We compared the internal colonization of healthy and symptomatic needles on two sites in Poland. In addition, the endophytic mycobiota in needles of various age was recorded on two other sites without disease symptoms. Fungi were isolated from 95.6% of the dead needles and from 62.9% of the living needles on symptomatic trees, whereas on healthy trees, only 45.0% of the needles were colonized internally. Colonization frequency increased with needle age. From a total of 2017 isolates, 116 fungal taxa were identified. Frequency of many species was influenced by needle type. Anthostomella formosa, Gloeosporidiella sp., Hypoxylon fragiforme, Xylaria hypoxylon and X. polymorpha were the most common fungi isolated from living needles. In symptomless living needles, fungi occurred significantly more often in the basal than in the apical parts. In dead needles, the most common fungi were Alternaria alternata, Paraconiothyrium sporulosum, Fusarium sp., Mollisia cinerea, Rhizoctonia sp., Rhizosphaera oudemansii, Thysanophora penicillioides, Xylaria hypoxylon and X. polymorpha. Rhizoctonia sp. was the most frequently isolated fungus in dead needles (23.4%) but occurred rarely also in living needles (0.3–1.1%). The supposed pathogen N. parasiticum was detected only sporadically (at most in 0.6% of the needles). Our findings demonstrate the need for understanding the role of Rhizoctonia sp. in Herpotrichia needle browning disease aetiology.     

Morphological and molecular characterization of Fusarium oxysporum and Fusarium avenaceum causing a novel needle blight of Picea asperata in China

Picea asperata is a unique tree species in China. It has important economic and ecological values. In this study, the disease symptoms resembling needle blight, a new disease of P. asperata (10- to 15-year-old), were detected in a forest area in Tianshui City, Gansu Province, China, in August 2020. Affected needles turned yellow, the colour became darker at the junction of infected and healthy areas, the needles wilted and even fell off the tree. Through pathogenicity tests, the two pure fungi strains isolated from diseased needles caused distinct needle blight symptoms on P. asperata, with symptoms similar to those observed in the forest. It was observed that they had sickle-shaped conidia. Subsequently, multilocus phylogenetic analysis of partial sequences of the internal transcribed spacer region (ITS), translation elongation factor 1-α (TEF-1α) and RNA polymerase II second major subunit (RPB2) genes was performed. They were found closely clustered with Fusarium oxysporum and F. avenaceum, respectively. Based on morphological and molecular biological determinations, the two pathogens were identified as F. oxysporum and F. avenaceum. To our knowledge, this is the first report of needle blight caused by F. oxysporum and F. avenaceum on P. asperata in China.     

The hosts and geographic range of Dothistroma needle blight in Slovakia

The occurrence and distribution of Dothistroma needle blight (DNB) were studied in 2014–2017 around Slovakia. A total of 84 localities, both native and planted, were investigated, and the presence of DNB was confirmed in 73 of them. In all positive locations, symptoms typical of DNB were observed and the Dothistroma species was confirmed using species‐specific primers either from fungal cultures or directly from needles. Both Dothistroma species—D. septosporum and D. pini—were identified. Both species occurred together in 29 locations, only D. septosporum in 42 and only D. pini in two locations. The host range of D. septosporum included 10 pine species and two spruce species. The host range of D. pini comprised the same number of pine hosts but only one spruce species. Five pine hosts, P. aristata, P. coulteri, P. densiflora, P. jeffreyi, P. × schwerinii, and one spruce host P. abies are new hosts species of D. pini. P. densiflora and Picea pungens have earlier been reported to be susceptible for DNB. In this study, D. septosporum was found from both tree species.     

Needle blight caused by Dothistroma pini in Slovakia: distribution,host range and mating types

Dothistroma needle blight (DNB) has been observed in Slovakia during the last two decades. Up until 2017, Dothistroma septosporum has only been detected and molecularly confirmed to cause DNB in Slovakia. Here, we report the detection of Dothistroma pini at six localities around Slovakia, representing different plantation types. Four pine species (Pinus sylvestris, P. nigra, P. mugo and P. jeffreyi) were confirmed as hosts of D. pini in Slovakia, of which only P. mugo has been previously reported as host in Slovakia. Three gene regions (ITS, EF1 –α, and ß-tubulin) of each of the 13 isolates were sequenced and assigned as D. pini. Based on ITS sequences, the studied isolates represent the haplotypes Dp_HAP.1, Dp_HAP.2. Both mating types were detected but at different localities. Our results suggest that in addition to D. septosporum, D. pini may contribute to DNB also in Slovakia.     

Bacterial shoot blight of sweet crab apple caused by Pseudomonas viridiflava

Sweet crab apple (Malus coronaria) is a popular ornamental tree planted in gardens and urban forests in South Korea. In 2018, severe symptoms were observed on street plantings of sweet crab apple in Jinju, South Korea including shoot blight: shoot tips, twigs and foliage of infected trees turned brown, and dried completely. The causal agent isolated from the disease lesions was identified as Pseudomonas viridiflava based on biochemical tests, pathogenicity, and 16S rRNA and rpoD gene sequencing. On artificial inoculation, this pathogen not only caused shoot blight on sweet crab apple, but also bulb rot and leaf blight on onion. Here, we report the occurrence of bacterial shoot blight caused by P. viridiflava in sweet crab apple for the first time in the world.     

Invasive brown spot needle blight caused by Lecanosticta acicola in Estonia

Lecanosticta acicola (Thüm.) Syd., a serious foliage pathogen of pines in many regions of the world, is an emerging invasive species in northern Europe. After the first record of L. acicola on Pinus ponderosa Dougl. ex Laws. in northern Estonia in 2008, monitoring was started to investigate the spread and host range of the fungus in the country. By the beginning of 2015, L. acicola was also recorded on P. uncinata Mill. ex Mirb., P. mugo Turra and on P. mugo var. pumilio (Haenke) Zenari, being the northernmost records of the fungus in Europe. So far, the single native pine species Pinus sylvestris L. has not been found to be infected. Molecular analysis proved infection of L. acicola on pines in five different localities of Estonia: in Tallinn (mainly in the Botanic Garden, northern Estonia), in Tori and Kärdla (western Estonia), in Vasula and Kärevere (east-central Estonia). The sexual state (teleomorph) of the fungus was not found, but the existence of the both mating types, MAT1-1 and MAT1-2, was confirmed.     

Providencia rettgeri as the causal agent of the brown slime flux of Populus tomentosa

Brown slime flux seeps slowly out of wounds and flows down the bark of roadside Populus tomentosa in Beijing, China. Two bacterial isolates, SL2‐2 and SL3‐3, obtained from the brown slime flux were identified as Providencia rettgeri based on 16S rRNA gene sequencing. After inoculation into the bark of roadside P. tomentosa with sterile deionized water as a control, both SL2‐2 and SL3‐3 triggered the seepage of brown slime flux from the wounds; no slime flux arose from the control. In conclusion, P. rettgeri was the pathogenic bacterium causing brown slime flux on P. tomentosa probably by inhibiting wound healing. To our knowledge, this is the first report of P. rettgeri as a pathogen of plants.     

Pathogenicity of Malaysian Phytophthora palmivora on cocoa,durian, rubber and oil palm determines the threat of bud rot disease

The epidemic of bud rot disease affecting oil palm in Colombia is primarily caused by Phytophthora palmivora. The pathogen has a cosmopolitan presence that includes Southeast Asia, but to date, bud rot has not been reported in this region. This study provides an overview of the potential risk of Malaysian P. palmivora isolates cross‐infecting other host species, including cocoa, durian, rubber and Malaysian oil palm planting materials (Dura × Pisifera, D × P). On cocoa pods, the durian isolate PP7 caused dark brown necrotic lesions. Detached leaf bioassays showed that P. palmivora isolates PP3 and PP7 infected different hosts, except rubber foliage without wounding. Inoculation tests on cocoa, durian and rubber seedlings caused brown necrotic lesions when stems were wounded, with 10% mortality in cocoa and durian at 17 days post‐inoculation (dpi). However, no further infection was observed, and lesions closed within 14–28 dpi on the non‐wounded seedlings. Pathogenicity tests of oil palm seedlings inoculated with isolates PP3 and PP7 indicated that Malaysian P. palmivora isolates were not pathogenic to oil palms based on localized infection observed only through wounding. Overall, the work demonstrated that Malaysian P. palmivora isolates were able to cross‐infect multiple hosts but did not show severe infections on oil palms.     

Analysis of fungal endophytes in Scottish Sitka spruce plantations shows extensive infections,novel host partners and gives insights into origins

This preliminary study identified the fungal endophytes of Sitka spruce (Picea sitchensis) in four Scottish plantations, analysed their recruitment possibilities and community structure across the sites, and identified the possible origins of the endophytes in this non‐native host. Although Sitka spruce is native to north‐western North America, it comprises a huge portion of Scottish forestry and is an economically vital timber tree. Needles from two age classes were collected and cultured for emerging endophyte isolates. Using a combination of morphological features and genetic sequencing of the nuclear ribosomal ITS regions of representative morphological types, 57 morphotypes were recorded which represented at least 15 different species, including a hypothesized sister species to the pathogen of Pseudotsuga menziesii in North America and Europe, Nothophaeocryptopus gaeumannii (previously P. gaeumannii) that was present at all sites tested. Diversity, recruitment, site, needle age and needle part effects are discussed. The phylogeny of Nothophaeocryptopus based on analysis of combined ITS and Tub2 sequence data is given in order to assess the relationship of the unknown endophyte to the Douglas fir pathogen N. gaeumannii.     

Occurrence of Diplodia corticola,including new oak host records,in Wisconsin,USA

In the United States, diseased oaks (Quercus species) exhibit tip blight, branch and stem cankers, and dieback often attributed to Diplodia species or related fungi. Emergence of Diplodia corticola as a pathogen of European oaks, and reports of this fungus in the eastern and western United States, prompted re‐examination of strains from Wisconsin. These had been obtained in the late 1990s and early 2000s and previously identified only as Diplodia species. Nuclear ribosomal ITS sequences were obtained from the Wisconsin strains and analysed with other sequences from GenBank. Wisconsin strains confirmed as D. corticola were from northern red oak (Q. rubra), black oak (Q. velutina), white oak (Q. alba) and bur oak (Q. macrocarpa). Other strains from oaks in Wisconsin were D. mutila and D. seriata. Wound inoculation of northern red, white and bur oak seedlings with D. corticola in a greenhouse resulted in shoot death and stem lesions, from which the pathogen was reisolated. We conclude that D. corticola has been present in the northcentral United States for at least two decades and report two previously unrecognized hosts of this pathogen: white oak and bur oak. The roles of D. corticola, related fungi and influences of other environmental factors in deterioration of oak health in North America merit additional investigation.