Search Results (27)

Dothistroma needle blight (DNB) is one of the most significant diseases of conifers, causing premature defoliation, growth reduction, and, in extreme cases, mortality. Histological analysis was undertaken on inoculated seedlings of three different seed sources of Pinus sylvestris L. to investigate the process of infection and degradation of needle tissue on this host species. Seedlings were inoculated using a single spore isolate of Dothistroma septosporum (Doroguine) M. Morelet (D636) from northern Scotland. Mesophyll degradation in the needles occurred by four weeks after inoculation; collapse of mesophyll, bundle sheath tissues, and tracheids by five weeks; and eruption of fruiting bodies in near proximity to stomatal openings by six weeks. Significantly greater collapse of mesophyll during the early stages of infection occurred in the Austrian provenance compared with the United Kingdom provenance, although in the later stages of infection, this difference disappeared. Furthermore, disease severity, assessed as the proportion of needles with D. septosporum conidiomata on each tree, was not significantly different between seed sources. Full article

Pine needle blight of Pinus massoniana caused by pathogens of the Pestalotiopsis genus is a destructive disease worldwide, especially in young forests. Chemical fungicides accelerate the formation of resistant strains among plant pathogenic fungi, which makes microbial biocontrol particularly important. In this study, we identified Neopestalotiopsis camelliae-oleiferae as a new pathogen of pine needle blight in P. massoniana via pathogen isolation, inoculation, pathogenicity assays, morphology observations, and multilocus phylogenetic analyses of the ITS, TEF1, and TUB2 regions. PSM-6, an endophytic bacterium, was subsequently isolated from pine needles and was shown to have excellent antagonistic activity against N. camelliae-oleiferae in vitro. Based on the morphology, physiology, and molecular analysis, we identified this strain as P. silvicola. The extracellular secondary metabolites of PSM-6 were further proven to cause the shrinkage and collapse of pathogen hyphae. The decreased disease index and mortality indicated that pretreatment with PSM-6 may effectively protect pine seedlings from pathogen infection. In addition, PSM-6 exhibited broad-spectrum antifungal activity in several phytopathogenic fungi, including Fusarium graminearum, Botrytis cinerea, and Verticillium dahliae. These findings establish PSM-6 as a promising biocontrol agent, offering an environmentally friendly alternative to chemical fungicides for managing pine needle blight and other fungal diseases. Full article

In June 2020, needle blight symptoms on Pinus thunbergii were discovered in Bazhong City, Sichuan Province, China. Fungal isolates were obtained from the pine needles of P. thunbergii. After examining morphological characteristics and conducting multi-locus (ITS, ACT, TUB2 and RPB2) phylogenetic analyses, the isolates SC1–SC5 were determined to be a new species, Biscogniauxia sinensis. Genealogical Concordance Phylogenetic Species Recognition with a pairwise homoplasy index test was used to further verify the results of the phylogenetic analyses. The morphology and phylogenetic relationships between this new species and other related Biscogniauxia species were discussed. To our knowledge, this is also the first report of Biscogniauxia sinensis associated with pine needle blight on P. thunbergii in China. The needle damage of P. thunbergii associated with Biscogniauxia sinensis will detrimentally affect the carbon absorption and photosynthetic efficiency of P. thunbergii, further reduce the absorption of nutrients by Japanese black pine and may lead to the imbalance of pine forest conditions, which will have a negative impact on the forest ecological system. Full article

Pinus massoniana Lamb. is an important, common afforestation and timber tree species in China. Species of Pestalotiopsis are well-known pathogens of needle blight. In this study, the five representative strains were isolated from needle blight from needles of Pi. massoniana in Nanjing, Jiangsu, China. Based on multi-locus phylogenetic analyses of the three genomic loci (ITS, TEF1, and TUB2), in conjunction with morphological characteristics, a new species, namely Pestalotiopsis jiangsuensis sp. nov., was described and reported. Pathogenicity tests revealed that the five representative strains of the species described above were pathogenic to Pi. massoniana. The study revealed the diversity of pathogenic species of needle blight on Pi. massoniana. This is the first report of needle blight caused by P. jiangsuensis on Pi. massoniana in China and worldwide. This provides useful information for future research on management strategies of this disease. Full article

Diplodia tip blight, caused by Diplodia sapinea (=Sphaeropsis sapinea), are widely distributed in Honghuaerji, Inner Mongolia, China, causing severe damage on natural Mongolian pine (Pinus sylvestris var. mongolica). D. sapinea is an endophyte that becomes pathogenic under conditions of drought, hail damage, or temperature-associated stress. The role of the endophytic community inhabiting different pine tissues in the expression of disease is still unknown. In this study, the diversity and community structure of endophytic fungi among asymptomatic and symptomatic Mongolian pine were detected using culture-based isolation and high-throughput sequencing (HTS), and the potential antagonistic endophytes against D. sapinea were also screened. The results indicated that 198 and 235 strains of endophytic fungi were isolated from different tissues of symptomatic and asymptomatic Mongolian pine, respectively. D. sapinea was the most common endophyte isolated from the current-year needles and shoots of symptomatic trees, and Diplodia was also the most common in the HTS data. There were no significant differences in the endophytic fungal species richness among asymptomatic and symptomatic trees, but there were differences observed within specific sampled tissues. The ANOSIM analysis confirmed that the endophytic fungi community structure significantly differed between sampling tissues among symptomatic and asymptomatic Mongolian pine. Furthermore, the antagonism study revealed Penicillium fructuariae-cellae with the ability to inhibit the growth of D. sapinea in vitro, and the potential performance of this fungus, acting as biological control agent, was evaluated under greenhouse. Our findings can pave the way to a better understanding of the interactions between D. sapinea, other endophytic fungi and their hosts, and provide helpful information for more efficient disease management strategies. Full article

Black spot needle blight is a minor disease in Mongolian Scots pine (Pinus sylvestris var. mongolica) caused by Pestalotiopsis neglecta, but it can cause economic losses in severe cases. Sodium pheophorbide a (SPA), an intermediate product of the chlorophyll metabolism pathway, is a compound with photoactivated antifungal activity, which has been previously shown to inhibit the growth of P. neglecta. In this study, SPA significantly reduced the incidence and disease index and enhanced the chlorophyll content and antioxidant enzyme activities of P. sylvestris var. mongolica. To further study the molecular mechanism of the inhibition, we conducted a comparative proteomic analysis of P. neglecta mycelia with and without SPA treatment. The cellular proteins were obtained from P. neglecta mycelial samples and subjected to a tandem mass tag (TMT)-labelling LC-MS/MS analysis. Based on the results of de novo transcriptome assembly, 613 differentially expressed proteins (DEPs) (p < 0.05) were identified, of which 360 were upregulated and 253 downregulated. The 527 annotated DEPs were classified into 50 functional groups according to Gene Ontology and linked to 256 different pathways using the Kyoto Encyclopedia of Genes and Genomes database as a reference. A joint analysis of the transcriptome and proteomics results showed that the top three pathways were Amino acid metabolism, Carbohydrate metabolism, and Lipid metabolism. These results provide new viewpoints into the molecular mechanism of the inhibition of P. neglecta by SPA at the protein level and a theoretical basis for evaluating SPA as an antifungal agent to protect forests. Full article

Fungal diseases such as root rot and leaf blight cause substantial losses in coastal pine (Pinus thunbergii Parl.) seedling production, which hinders afforestation/forest restoration programs. We isolated and identified Fusarium oxysporum and Alternaria alternata as the causal agents of root rot and needle blight diseases and investigated the biocontrol efficacy against the fungal pathogens and growth promotion of coastal pine seedlings using Bacillus velezensis CE 100. The bacterium produced the hydrolytic enzymes chitinase, β-1,3-glucanase, and protease enzymes, and the crude enzyme fraction of the biocontrol strain caused the deformation of the fungal cell wall and antagonized F. oxysporum and A. alternata, causing respective inhibition of spore germination by 91.0% and 85.9% and mycelial growth by 58.3% and 54.3%, at a concentration of 1000 µL/mL. Consequently, the bacterial treatment improved the survival rate of seedlings 1.9 times relative to the control group. The bacterium secreted indole-acetic acid (IAA) phytohormone and enhanced root growth and absorption of nutrients, which notably enhanced the biomass production of coastal pine seedlings. Therefore, these results provide evidence that B. velezensis CE 100 is an effective antifungal and growth-promoting bacterium that can facilitate the production of high-quality coastal pine seedlings for the restoration and establishment of coastal forests. Full article

The international plant trade results in the accidental movement of invasive pests and pathogens, and has contributed significantly to recent range expansion of pathogens including Dothistroma septosporum. Seeds are usually thought to present a lower biosecurity risk than plants, but the importation of Pinus contorta seeds from North America to Britain in the mid-1900s, and similarities between British and Canadian D. septosporum populations suggests seeds could be a pathway. Dothistroma septosporum has not been isolated from seeds, but inadequately cleaned seed material could contain infected needle fragments. This case study investigated whether cone kilning, and wet and dry heat treatments could reduce D. septosporum transmission without damaging seed viability. Pinus needles infected with D. septosporum were incubated alongside cones undergoing three commercial seed extraction processes. Additional needles were exposed to temperatures ranging from 10 to 67 °C dry heat for up to 48 h, or incubated in water heated to between 20 and 60 °C for up to one hour. Pinus sylvestris seeds were exposed to 60 and 65 dry heat °C for 48 h, and further seed samples incubated in water heated to between 20 and 60 °C for up to one hour. Dothistroma septosporum survived the three kilning processes and while seeds were not damaged by dry heat exceeding 63.5 °C, at this temperature no D. septosporum survived. Wet heat treatments resulted in less than 10% pathogen survival following incubation at 40 °C, while at this temperature the seeds suffered no significant impacts, even when submerged for one hour. Thus, commercial seed kilning could allow D. septosporum transmission, but elevated wet and dry heat treatments could be applied to seed stock to minimise pathogen risk without significantly damaging seed viability. Full article

Lecanosticta acicola is one of the most damaging species affecting Pinus radiata plantations in Spain. Favourable climatic conditions and unknown endogenous factors of the pathogen and host led to a situation of high incidence and severity of the disease in these ecosystems. With the main aim of understanding the factors intrinsic to this pathogenic species, a study of the population structure in new established plantations with respect to older plantations was implemented. The genetic diversity, population structure and the ability of the pathogen to spread was determined in Northern Spain (Basque Country), where two thirds of the total Pinus radiata plantations of Spain are located. From a total of 153 Lecanosticta acicola isolates analysed, two lineages were present; the southern lineage, which was prevalent, and the northern lineage, which was scarce. A total of 22 multilocus genotypes were detected with a balanced composition of both mating types and evidence for sexual reproduction. In addition to the changing environmental conditions enhancing disease expression, the complexity and diversity of the pathogen will make it difficult to control and to maintain the wood productive system fundamentally based on this forest species. Full article

Black spot needle blight is a serious conifer disease of Pinus sylvestris var. mongolica occurring in Northeast China, which is usually caused by the plant pathogenic fungus Pestalotiopsis neglecta. From the diseased pine needles collected in Honghuaerji, the P. neglecta strain YJ-3 was isolated and identified as the phytopathogen, and its culture characteristics were studied. Then, we generated a highly contiguous 48.36-Mbp genome assembly (N50 = 6.62 Mbp) of the P. neglecta strain YJ-3 by combining the PacBio RS II Single Molecule Real Time (SMRT) and Illumina HiSeq X Ten sequencing platforms. The results showed that a total of 13,667 protein-coding genes were predicted and annotated using multiple bioinformatics databases. The genome assembly and annotation resource reported here will be useful for the study of fungal infection mechanisms and pathogen–host interaction. Full article

Pinus bungeana, an endangered and native coniferous tree species in China, has considerable timber and horticulture value. However, little is known about needle diseases in P. bungeana. A needle blight of P. bungeana has been observed in Hebei Province, China. P. bungeana inoculated with mycelial plugs of fungal isolates presented symptoms similar to those observed under field conditions. Ten virulent fungal isolates were identified as a small-spored Alternaria species based on morphological observations. Maximum likelihood and Bayesian phylogenetic analyses carried out with multilocus sequence typing of eight regions (SSU, LSU, ITS, gapdh, tef1, Alt a 1, endoPG, OPA10-2) assigned the pathogen to Alternaria alternata. This is the first report of A. alternata causing needle blight on P. bungeana in China. Full article

Dothistroma needle blight, caused by Dothistroma septosporum, has increased in incidence and severity over the last few decades and is now one of the most important global diseases of pines. Disease resistance breeding could be accelerated by knowledge of pathogen virulence factors and their host targets. However, this is hindered due to inefficient targeted gene disruption in D. septosporum, which is required for virulence gene characterisation. Here we report the first successful application of CRISPR/Cas9 gene editing to a Dothideomycete forest pathogen, D. septosporum. Disruption of the dothistromin pathway regulator gene AflR, with a known phenotype, was performed using nonhomologous end-joining repair with an efficiency of >90%. Transformants with a range of disruption mutations in AflR were produced. Disruption of Ds74283, a D. septosporum gene encoding a secreted cell death elicitor, was also achieved using CRISPR/Cas9, by using a specific donor DNA repair template to aid selection where the phenotype was unknown. In this case, 100% of screened transformants were identified as disruptants. In establishing CRISPR/Cas9 as a tool for gene editing in D. septosporum, our research could fast track the functional characterisation of candidate virulence factors in D. septosporum and helps set the foundation for development of this technology in other forest pathogens. Full article

Foliage samples from Podocarpus totara with severe needle browning and needle loss in the lower part of the crown were observed in 2011 in the Gisborne region of New Zealand. A Phytophthora genus-specific test applied directly to the needles gave a strong positive result, and subsequent isolations yielded colonies of a slow-growing oomycete. Morphological examination in vitro revealed a Phytophthora species. Preliminary comparisons of the rDNA (ITS), and ras-related protein (Ypt) gene regions with international DNA sequence revealed low sequence similarity to species from the downy mildew genus Peronospora, as well as clade 3 Phytophthora species. Other studies have also demonstrated the close relationship with Peronospora. The species was given the interim designation Phytophthora taxon tōtara pending further examination. Here, we formally describe Phytophthora podocarpi sp. Nov. and its associated disease, tōtara needle blight. Full article

In the last decade, the impact of needle blight fungal pathogens on the health status of forests in northern Spain has marked a turning point in forest production systems based on Pinus radiata species. Dothistroma needle blight caused by Dothistroma septosporum and D. pini, and brown spot needle blight caused by Lecanosticta acicola, coexist in these ecosystems. There is a clear dominance of L. acicola with respect to the other two pathogens and evidence of sexual reproduction in the area. Understanding L. acicola spore dispersal dynamics within climatic determinants is necessary to establish more efficient management strategies to increase the sustainability of forest ecosystems. In this study, spore counts of 15 spore traps placed in Pinus ecosystems were recorded in 2019 and spore abundance dependency on weather data was analysed using generalised additive models. During the collection period, the model that best fit the number of trapped spores included the daily maximum temperature and daily cumulative precipitation, which was associated to higher spore counts. The presence of conidia was detected from January and maximum peaks of spore dispersal were generally observed from September to November. Full article

In recent years, the decline of pine stands in Europe, including Poland, has been caused by the emerging needle pathogen Dothistroma septosporum. Although this fungus appears to preferentially infect Pinus pini, P. pinaster or P. radiata in Southern Europe, it has been reported in stands of P. nigra, P. mugo and P. sylvestris from Southern Poland. Our preliminary tests of symptomatic needles of diseased pines, including black pine (P. nigra), showed the presence of both D. septosporum and D. pini—the latter as the first report in Poland. No other endophytic pathogen, i.e., Lecanosticta acicola or Cenangium ferruginosum, were found. More extensive molecular surveying based on β-tub2 amplification of DNA in needle samples from 72 seed trees of P. sylvestris in nine different Forest Districts of Southern Poland did not find the presence of D. septosporum. Our study revealed that the seed trees from which we collected propagation material were free from the pathogen, and its endophytic behavior was not confirmed in our testing. Consequently, these investigated trees of P. sylvestris should be suitable for seed collection and propagation, following the requirements of “good” phytosanitary quality as “pathogen-free” pine seeds used for reforestation. Full article