Two new root endophyte and nematode cyst parasite species of the widely distributed genus Laburnicola

Fungal root endophytes, including the common form group of dark septate endophytes (DSEs), represent different taxonomic groups with potentially diverse life strategies. During surveys of DSE communities and of nematode cysts colonizing fungi, isolates representing Laburnicola ( Didymosphaeriaceae , Pleosporales ) lineages were discovered. Here we carried out a comprehensive study of the phylogenetic relationships and taxonomy of fungi collected from plant roots in Hungary, Mongolia, and Kazakhstan and from eggs of the cereal cyst nematode Heterodera filipjevi in Turkey. In addition to the study of the morphology and culture characteristics of the strains, four loci (internal transcribed spacer, partial large and small subunit regions of nuclear ribosomal DNA and partial translation elongation factor 1-alpha) were used to infer the molecular phylogenetic relationships of the strains within Laburnicola . The isolates were found to represent two distinct lineages, which are described here as novel species, Laburnicola nematophila and L. radiciphila . The interaction of the strains with plants and nematodes was examined using in vitro bioassays, which revealed endophytic interactions with the plant roots and parasitic interactions with the nematode eggs. Analyses of similar ITS sequences found in public databases revealed that members of the genus Laburnicola are widely distributed characteristic members of the plant microbiome, and they are reported as parasites of plant parasitic cyst nematodes here for the first time

Neofabraea kienholzii, a novel causal agent of grapevine trunk diseases in Hungary

Recently, more and more new fungal pathogens have been described as causal agents of grapevine trunk diseases (GTDs), which lead to increasingly significant economic losses in viticulture worldwide. The genus Neofabraea consists of species mainly known as important plant pathogens causing perennial canker and bull’s eye rot, a common postharvest disease of apple (Malus domestica) and pear (Pyrus communis) fruits. Neofabraea kienholzii also causes lesions on pome fruits and canker on woody tissues, but its pathogenicity has not been demonstrated on grapevine yet. In 2015, two strains, identified as N. kienholzii based on ITS sequence data, were isolated from vines showing symptoms of GTDs in Hungary. For an unambiguous taxonomic placement of the isolates, four loci (ITS, LSU, TUB2 and RPB2) were amplified and sequenced. The phylogenetic analysis confirmed that the two isolates represent N. kienholzii. Pathogenicity tests performed on potted grapevines, shoots, and canes confirmed the virulence of these fungi. Their growth and sporulation on different media were also investigated. To the best of our knowledge, this is the first proof of N. kienholzii might cause symptoms on Vitis and might have a role in GTDs

Micro-scale Experimental System Coupled with Fluorescence-based Estimation of Fungal Biomass to Study Utilisation of Plant Substrates.

The degradation capacity and utilisation of complex plant substrates are crucial for the functioning of saprobic fungi and different plant symbionts with fundamental functions in ecosystems. Measuring the growth capacity and biomass of fungi on such systems is a challenging task. We established a new micro-scale experimental setup using substrates made of different plant species and organs as media for fungal growth. We adopted and tested a reliable and simple titration-based method for the estimation of total fungal biomass within the substrates using fluorescence-labelled lectin. We found that the relationship between fluorescence intensity and fungal dry weight was strong and linear but differed among fungi. The effect of the plant organ (i.e. root vs. shoot) used as substrate on fungal growth differed among plant species and between root endophytic fungal species. The novel microscale experimental system is useful for screening the utilisation of different substrates, which can provide insight into the ecological roles and functions of fungi. Furthermore, our fungal biomass estimation method has applications in various fields. As the estimation is based on the fungal cell wall, it measures the total cumulative biomass produced in a certain environment

Root-Colonizing Endophytic Fungi of the Dominant Grass Stipa krylovii From a Mongolian Steppe Grassland

In several terrestrial ecosystems such as grasslands, plants live together with various root-colonizing dark septate endophytes (DSEs), fungi that are relatively frequent colonizers of healthy belowground tissues of plants in these environments. They are important members of the plant microbiota and may have various effects on plant survival under different stress conditions; however, their general functions in relation to plants and the greater ecosystem remain elusive. Although an increasing number of studies has been published focusing on DSEs in Asian grasslands, our knowledge is limited. Especially in Mongolia, where the steppe region represents a significant area, information is not available on these root colonizers. In this study, we aimed to characterize DSEs of a common dominant gramineous plant species, Stipa krylovii in a semiarid grassland of Mongolia. Root samples were collected in a natural steppe and were processed for isolation of fungal endophytes. For molecular identification of the isolates, the internal transcribed spacer (ITS) region of the nrDNA was obtained for all the isolates investigated; furthermore, the partial translation elongation factor 1-a (TEF) gene and large subunit (LSU) and small subunit (SSU) of rDNA were also amplified and sequenced in case of representative isolates. In vitro tests were used to examine the rough symbiotic nature of the fungi, and root colonization was visualized. A majority of the 135 isolates examined in detail was found to belong to several orders of Ascomycota (110 isolates) and some to Basidiomycota (25 isolates). A significant number of the isolates represented presumably novel taxa, and dominant similarities of the lineages have been found with relatively frequent and known grass root endophytes of semiarid areas in other geographic regions. These endophytes included Periconia macrospinosa, Microdochium bolley, and Darksidea, the genus of which comprised one fourth of the isolates. We found numerous lineages, which have been detected not only from Asian steppe ecosystems, but also from prairies in North America and sandy grasslands in Europe. Therefore, our results strengthen the hypothesized worldwide presence of a common and dominant core group of a DSE community in arid and semiarid grasslands