Sphaeropsis sapinea is the causal fungal agent of Diplodia tip blight disease of Scots pine (Pinus sylvestris) and other coniferous trees of relevance to forestry in Germany. In this study, the distribution and occurrence of S. sapinea and accompanying endophytic fungi in twigs of healthy and diseased Scots pine was investigated on a spatial and temporal scale. Sampling of 26,000 twig segments from trees in 105 temperate coniferous forest stands in Germany resulted in isolation of 33,000 endophytic fungi consisting of 103 species identified based on morphological and ITS-DNA sequence analyses. Approximately 98% of the sample was represented by fungi in the Ascomycota, with only two species (Peniophora pini and Coprinellus sp.) belonging to the Basidiomycota. Four species were detected in a frequency greater than 10% (Sphaeropsis sapinea, Sydowia polyspora, Microsphaeropsis olivacea, and Truncatella conorum-piceae) from the collective sample. A typical inhabitant of Scots pine twigs Desmazierella acicola was isolated and additionally typical hardwood colonizers like Biscogniauxia spp. were detected. S. sapinea, an endophytic plant pathogen with saprobic capabilities, was isolated from more than 80% of the studied pine trees, but the majority of trees sampled showed no symptoms of Diplodia tip blight. No invasive, pathogenic quarantine fungi for Germany were isolated from healthy or diseased Scots pines. Advantages and disadvantages of isolation-based endophyte studies over studies using direct DNA-isolation are discussed. Knowledge of the fungal endophyte communities in twigs of Scots pine allowed for identification S. sapinea and other potential pathogens of pines and other forest trees that may possibly contribute to increased disease under repeated periods of drought and heat stress in the future.
The ascomycete Sphaeropsis sapinea is the causal agent of the Diplodia Tip Blight disease on pines and other conifer species. This fungus has a symptomless endophytic life stage. Disease symptoms become visible when trees have been weakened by abiotic stress, usually related to warmer temperatures and drought. Currently, this disease is observed regularly in Scots pine (Pinus sylvestris) sites in parts of Europe, such as Germany, increasing dramatically in the last decade. Changes in climatic conditions will gradually increase the damage caused by this fungus, because it is favored by elevated temperature. Thus, host trees with reduced vitality due to climate change-related environmental stress are expected to be more susceptible to an outbreak of Diplodia Tip Blight disease. There is currently no established and effective method to control S. sapinea. This project aims to reveal the nature of the endophyte community of Scots pine. Utilizing the antagonistic core community of endophytes could serve as a novel tool for disease control. Results from this study provide a starting point for new solutions to improve forest health and counter S. sapinea disease outbreaks. We screened potential antagonistic endophytes against S. sapinea and infected Scots pine seedlings with the most common endophytes and S. sapinea alone and combination. The host was stressed by limiting access to water. The antagonism study revealed 13 possible fungi with the ability to inhibit the growth of S. sapinea in vitro, for example Sydowia polyspora. None of the tested co-infected fungi (Desmazierella acicola, Didymellaceae sp., Microsphaeropsis olivacea, Sydowia polyspora, and Truncatella conorum-piceae) showed strong necrosis development in vivo, even when host stress increased due to drought. However, the infection experiment demonstrated that drought conditions enhance the effect of the disease outbreak, triggering S. sapinea to cause more necrosis in the infected twigs.
Research Highlights: Dedicated experiments to investigate how disturbances will affect Heterobasidion sp.—Norway spruce pathosystems are important, in order to develop different strategies to limit the spread of Heterobasidion annosum s.l. under the predicted climate change. Here, we report on a greenhouse experiment to evaluate the effects of water availability on the infection severity of Heterobasidion parviporum or Heterobasidion annosum, respectively, on Picea abies saplings. Background and Objectives: Changes in climatic conditions and intense logging will continue to promote H. annosum s.l. in conifer forests, increasing annual economic losses. Thus, our aim was to test if disease severity in Norway spruce was greater after infection with H. parviporum or H. annosum in low water availability conditions, compared to seedlings with high water availability. Materials and Methods: We performed inoculation studies of three-year-old saplings in a greenhouse. Saplings were treated as high (+) or low (−) water groups: High water group received double the water amount than the low water group. The necrosis observed after pathogen inoculation was measured and analyzed. Results: The seedling growth was negatively influenced in the lower water group. In addition, the water availability enhanced the necrosis length of H. parviporum in phloem and sapwood (vertical length) in the low water group. H. annosum benefited only in horizontal length in the phloem. Conclusions: Disturbances related to water availability, especially low water conditions, can have negative effects on the tree host and benefit the infection ability of the pathogens in the host.
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