Materiały konferencyjne SEP 2024

2 environmental conditions such as high soil pH [11], low nutrient availability [12] and toxic metal concentrations that limit microbial growth. The limited availability of nutrients in mine dumps also affects plant growth and makes it difficult to establish a vegetation cover [13]. To date, most studies on plant-microbe interactions in mine dumps have focused on the effects of plant-microbe interactions on vegetation cover establishment [14], while less attention has been paid to microbial community structure and activity. However, recent studies have shown that microbial communities play a crucial role in ecosystem restoration by facilitating vegeta- tion cover establishment and nutrient cycling [15]. Therefore, understanding the structure and activity of microbial communities on mine tailings is crucial for the development of effective remediation strategies. The soil will be characterized in terms of physiochemical parameters and respiration. Substrate pH will be measured with a glass electrode and the electrical conductivity. The organic carbon content of the soil will be determined using the Turin method modified by Simakov [16]. Total nitrogen will be determined using the Kjeldahl method [17]. The content of available forms of phosphorus will be determined using the Egner-Riehm method. The concentration of magne- sium extracted using the Schachtschabel method [18] and the exchangeable potassium and so- dium cations will be determined by flame absorption spectrometry. Substrate moisture will be determined in situ using the portable soil moisture sensor ML3 ThetaKit and substrate respira- tion will be determined using a portable infrared gas analyzer connected to a soil respiration chamber. The microbial community will be characterised by determining the microbial biomass, micro- bial community structure and microbial community activity. The microbial biomass will be determined using qPCR and standard primers for the 16S rRNA gene for bacteria [19], the mcrA gene for archaea and the ITS region for fungi and oomycetes. Microbial structure will be investigated using a protocol for phospholipid fatty acid analysis [15] and next-generation sequencing. Amplicon sequencing on the MiniSeq® sequencer will be performed using pri- mers flanking the hypervariable regions of the 16S rRNA gene for bacteria, the mcrA gene for archaea, and the ITS region for fungi and oomycetes. The raw sequencing reads will be proces- sed with the DADA2 package [20] in R according to the standard operating procedure with the addition of reads concatenation [21]. Based on the representation of individual bacterial spe- cies, bacterial community activity will be predicted using PICRUSt 2.0 software [22]. The bio- chemical potential of the soil microorganisms will be determined using the Biolog EcoPlateTM system. Selected samples will be subjected to metatranscriptomic analysis to assess the activity of the microbial community at the transcriptional level. The experimental design of the entire experiment is shown in Figure 1. The expected impact of this research, with potential applications in the restoration of degraded ecosystems, is significant as it will contribute to the understanding of the relationships between plants and microbes in natural and novel ecosystems. The knowledge gained from this study can also inform the development of sustainable land management practices that can improve ecosystem functions and promote biodiversity conservation. In addition, this study can provide insights into the microbial processes involved in carbon and nutrient cycling in soils, which in turn has implications for climate change mitigation and adaptation.