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This paper continues the studies on the role of bacteria in the environment of summer truffle (Tuber aestivum), focusing on ascomata at three sites in the Nida Basin (southern Poland). Bacterial taxa and the number of OTU sequences identified were determined based on analysis of peridium and gleba collected in 2017 and 2018. The aim of the presented work is to learn the structure of bacterial communities associated with the ascomata of the summer truffle T. aestivum forming in specific stands in south Poland and fine roots of surrounding trees. Sanger DNA sequencing, and next-generation sequencing (NGS) were used. NGS sequencing platforms short-read (300 bases) was selected, using a MiSeq Illumina. The Kruskall-Wallis test was used for alpha diversity analysis and the PERMANOVA test for beta diversity. Alpha and beta biodiversity analyzes were performed using various indices (Chao1, Simpson, Shannon, Faith's phylogenetic, observed OTUs, Heip's evenness measure - alpha diversity; UniFrac, Jaccard, Bray–Curtis - beta diversity). The Sanger method identified six bacterial taxa that were dominant in the community: Bacillus (86.7 %), Pseudomonas (51.5 %), Stenotrophomonas (15.5 %), Paenibacillus (7.9 %), Rhizobium (7.3 %), and Chryseobacterium (8.5 %). In contrast, NGS results showed the dominance (at OTUs >10,000) of taxa: Bradyrhizobiaceae, Sphingobacteriaceae, Micrococcaceae, Yersiniaceae, Sphingomonadaceae, Rhizobiaceae, Flavobacteriaceae, and Comamonadaceae. The PERMANOVA test for the Bray-Curtis and Jaccard indices showed dissimilarities between samples from two of the sites with respect to beta diversity. Significant differences were found in the bacterial communities colonizing the ascomata of T. aestivum and in the root communities with T. aestivum ectomycorrhizae the purpose of this study. The results obtained may be helpful in selecting and monitoring sites suitable for the formation of ascomata of T. aestivum. [ABSTRACT FROM AUTHOR]

In this study, we examined the effect of the presence of mycorrhiza and ascomata of summer truffle (Tuber aestivum) on the bacterial composition of roots from small trees growing in selected sites of the Nida Basin. Qualitative DNA sequencing methods such as Sanger and next-generation sequencing (NGS) were used. The Sanger method revealed different bacterial species compositions between the samples where summer truffle ascomata was recorded and control samples. Five genera of bacteria could be distinguished: Bacillus, Erwinia, Pseudomonas, Rahnella and Serratia, among which the most numerous were Pseudomonas (Gammmaproteobacteria class) at 32.9%. The results obtained by the NGS method also showed differences in species composition of the bacteria depending on the study sample. Seven genera of bacteria were distinguished: Rhizorhabdus, Methylotenera, Sphingomonas, Nitrosospira, Streptomyces, Methyloceanibacter and Niastella, which dominated in roots from the truffle sites. Telmatobacter, Roseiarcus, Granulicella, Paludibaculum, Acidipila, Acidisphaera and Aliidongia dominated in roots from the control sites. With the NGS method, it is possible to identify the microbiome of a whole root, while only a root fragment can be analysed by the Sanger method. These results extend the scope of knowledge on the preferences of certain groups of bacteria associated with truffles and their influence on the formation of ascomata in summer truffles. Our results may also be useful in selecting and monitoring sites that promote ascomata of Tuber aestivum. [ABSTRACT FROM AUTHOR]

This paper describes the quantitative and qualitative composition of bacteria isolated from soil in the selected sites in the Nida Basin, in places where mycorrhizae and ascocarps of summer truffle (Tuber aestivum) were found, and in a control soil (without truffle). A classic growth culture method was used with Sanger DNA sequencing to obtain quantitative and qualitative measures of bacterial cultures. The obtained results showed differences in bacteriome composition between the case samples, in which summer truffle fructification was observed, and the control samples. Seven classes of bacteria were identified: Actinobacteria, Bacilli, Deinococci, Flavobacteria, Alphaproteobacteria, Betaproteobacteria, and Gammaproteobacteria. The most numerous bacterial genera were Pseudomonas (class Gammaproteobacteria) – 33%, Streptomyces (class Actinobacteria) – 29% and Bacillus (class Bacilli) – 15%. This research broadens the understanding of individual groups of bacteria accompanying truffles and their potential impact on the formation of summer truffle ascocarps. [ABSTRACT FROM AUTHOR]