Colonization of Fungi and Bacteria in Stumps and Roots of Scots Pine after Thinning and Treatment with Rotstop

The research areas were located in the Pisz Forest District, northeast Poland, in 10‐year‐old Scots pine (Pinus sylvestris L.) plantations, established in 2004 on a clear‐cut area. Reforestation was performed without a biological treatment against root pathogens, despite the presence of Heterobasidion annosum and Armillaria ostoyae in roots and stumps of trees growing previously. The aim of this research was to evaluate how thinning and treatment with the biological control agent Rotstop influences bacterial and fungal communities within roots and stumps. Twelve months after thinning, samples were collected from five stumps in each of two seasons, autumn and spring, from stands on two types of site, one previously forested and one agricultural (20 stumps in total). Wood samples were cultured on agar media, and (i) fungi in the upper part of the stump and (ii) in roots and (iii) bacteria in roots were genetically identified. Sequences were genetically identified by comparing sequences with records held in the GenBank database. We found great differences in the frequency of both fungi and bacteria in roots: they were more frequent (i) in healthy stumps compared to stumps infected with pathogens (H. annosum and A. ostoyae), (ii) in postagricultural soil than in forest soil and (iii) after spring rather than autumn biological treatment. The introduced species Phlebiopsis gigantea was only identified in the parts of the stumps which were above ground level. The bacterium Paenibacillus pini was associated with the presence of H. annosum infecting the stumps from the roots side. In areas seriously threatened by root pathogens, biological treatment can play only a limited role. It can spread to the upper part and impede the production of fruitbodies; however, it has no impact on the development of pathogens in deeper root areas.     

Impact of biological (Rotstop) and chemical (urea) treatments on fungal community structure in freshly cut Picea abies stumps

To control the infections by root rot fungi Heterobasidion spp., surfaces of freshly cut Norway spruce stumps are covered either by a biological (Rotstop; spore suspension of competitive saprotrophic fungus Phlebiopsis gigantea), or by a chemical (35% aqueous solution of urea) compound. In Fennoscandia, Rotstop and urea are applied, respectively, on 47,000 ha and on 2000 ha of forestland each year. The aim of this work was to assess the impact of biological and chemical control on biodiversity in communities of non-target fungi in freshly cut (7-week-old) stumps. Isolation of fungi to pure culture was accomplished from 402 wood samples taken from 63 stumps, 21 treated with each of the compounds and 21 untreated. The isolations yielded 368 distinct fungal strains representing 47 species. Stump treatment led to decrease of species richness both in Rotstop-treated (by 15%) and in urea-treated (by 19%) stumps. Nevertheless, the stumps subjected to the biological compound were colonized mainly by the same fungi that occurred naturally in untreated stumps (Sorensen similarity indices; S_S=0.69; S_N=0.68). By contrast, chemical treatment strongly promoted stump colonization by Ascomycetes and Deuteromycetes, led to significant decrease of Zygomycetes, and almost completely eliminated Basidiomycetes (including Heterobasidion spp.). Thus, resemblance to a natural community was low (S_S=0.45; S_N=0.34). Rotstop treatment decreased significantly the extent of stump colonization by Heterobasidion spp., and increased that of P. gigantea. All strains of the latter were genetically identical among themselves and to the Rotstop strain. The mechanisms of biological and chemical control, and biodiversity aspects are discussed.     

The capacity in Heterobasidion annosum s.l. to resist overgrowth by the biocontrol agent Phlebiopsis gigantea is a heritable trait

The necrotrophic pathogen Heterobasidion annosum sensu lato (Fr.) Bref. causes severe root rot on coniferous trees in the boreal and temperate forests. The annual economic losses caused by this fungus in Europe are estimated to at least 790 million €. In managed forests, the major route of infection is via stump surfaces from which the H. annosum s.l. grows through the roots and attacks adjacent healthy trees. A biocontrol method to reduce H. annosum s.l. infection is to apply the wood degrading fungus Phlebiopsis gigantea in a spore solution (Rotstop) directly on the freshly cut stumps immediately after cutting. We investigated the potential risk for a build-up in the capacity of H. annosum s.l. to resist overgrowth by P. gigantea. Wood blocks of Picea abies, precolonized with the two fungal species, were juxtaposed on top of agar and the overgrowth of the P. gigantea strain (Rotstop) on the H. annosum s.l. was measured periodically. We found a natural variation in Heterobasidion parviporum to resist overgrowth by P. gigantea. There was no difference between homo- and heterokaryotic strains. In a mapping population of 91 progenies from a H. annosum hybrid strain we were able to identify one quantitative trait locus (QTL) which controls the examined resistance capacity. We estimated the broad sense heritability to 0.336 for the capacity to resist the P. gigantea overgrowth. We conclude that there exists a theoretical risk for resistance build-up in the H. annosum s.l. population towards its biological control agent P. gigantea.