Soil enzyme activities (arylsulfatase, beta-glucosidase, alkaline and acid phosphomonoesterase, and protease activities) in off-trail samples (blank bar), inside the ruts samples (black bar), and between the ruts samples (grey bar), at T1 (a) and at T2 (b). The error bars indicate the standard deviation. For each enzymatic activity, different letters indicate statistical differences among different levels of soil compaction at each sampling time based on the Tukey test (P < 0.05).

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... the ruts (Fig. 2). Also, the protease activity was significantly lower inside the ruts compared to both off-trail and between the ruts soils ( Fig. 2). At T2 no significant differences were observed in the alkaline phosphomonoesterase, arylsulfatase and protease activities, while the acid phosphomonoesterase activity was significantly higher inside the ...

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... the ruts (Fig. 2). Also, the protease activity was significantly lower inside the ruts compared to both off-trail and between the ruts soils ( Fig. 2). At T2 no significant differences were observed in the alkaline phosphomonoesterase, arylsulfatase and protease activities, while the acid phosphomonoesterase activity was significantly higher inside the ruts than off-trail and between the ruts, showing contrasting results compared with the results at T1 (Fig. 2). The β-glucosidase ...

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... and between the ruts soils ( Fig. 2). At T2 no significant differences were observed in the alkaline phosphomonoesterase, arylsulfatase and protease activities, while the acid phosphomonoesterase activity was significantly higher inside the ruts than off-trail and between the ruts, showing contrasting results compared with the results at T1 (Fig. 2). The β-glucosidase activity inside and between the ruts was significantly lower than the offtrail at both sampling times (Fig. ...

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... and protease activities, while the acid phosphomonoesterase activity was significantly higher inside the ruts than off-trail and between the ruts, showing contrasting results compared with the results at T1 (Fig. 2). The β-glucosidase activity inside and between the ruts was significantly lower than the offtrail at both sampling times (Fig. ...

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... of 143,448 ± 33,811 ITS sequences per sample. After quality filtering, denoising, and removal of chimeras, approximately 297,113 high-quality 16S rRNA sequences, and 70,185 ITS sequences remained for the following analysis. The sequencing depths were enough to accurately describe the biodiversity within the bacterial and fungal communities (Fig. S2). As a result, the 16S rRNA sequences obtained were classified into 4675 ASVs assigned to both Archaea and Bacteria Kingdoms, 37 phyla, 81 classes, 163 orders, 209 families, and 287 genera. Instead, ITS sequences resulted in a total of 1422 ASVs, 14 phyla, 32 classes, 63 orders, 88 families, and 135 genera. A complete list of the ...

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... Planctomycetota (4 % -6 %), Chloroflexi (1 % -3 %) and Firmicutes (2 % -3 %) (Fig. S2 A). Alphaproteobacteria was the most abundant class of Proteobacteria with an average relative abundance between ~16 % and ~21 %, followed by Gammaproteobacteria, which accounted for a range of mean relative abundance from 9 % to 14 ...

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... order and genus level at each sampling time and entirely reported in File S2. In particular, at sampling time T1 shifts of the order relative abundances among inside and between the ruts samples were observed. The orders Acidobacteriales, Subgroup 2, and WD260 were significantly more abundant inside the ruts compared to between the ruts samples (Fig. 3A, File S2). Differently, Sphingomonadales and Vicinamibacterales were significantly less abundant inside the ruts than between the ruts samples (Fig. 3A, File S2). Additionally, the order Saccharimonadales were significantly more present in compacted soils (between and inside the ruts samples) than in the undisturbed soils (off-trail samples) ...

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... among inside and between the ruts samples were observed. The orders Acidobacteriales, Subgroup 2, and WD260 were significantly more abundant inside the ruts compared to between the ruts samples (Fig. 3A, File S2). Differently, Sphingomonadales and Vicinamibacterales were significantly less abundant inside the ruts than between the ruts samples (Fig. 3A, File S2). Additionally, the order Saccharimonadales were significantly more present in compacted soils (between and inside the ruts samples) than in the undisturbed soils (off-trail samples) (File S2). Moreover, the post hoc comparisons performed at sampling time T1 showed that genera Gemmatimonas and Sphingomonas were significantly less ...

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... the bacterial genera, 15 genera showed an average relative abundance of 1 %, or higher, in at least one group of soil conditions tested (Fig. 3C, File S2): the most abundant genera were Candidatus Solibacter (average relative abundance ranged from 3 to 6 % approximately), Candidatus Udaeobacter (~3 % -~6 %), Bryobacter (~3 % -~6 %), Puia (~1 % -~3 %), Acidothermus (~1 % -~2 %) and Streptococcus (~1 % -~2 %) (Fig. 3C, File S2). The bacterial genera profile in inside the ruts was deeply ...

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... of 1 %, or higher, in at least one group of soil conditions tested (Fig. 3C, File S2): the most abundant genera were Candidatus Solibacter (average relative abundance ranged from 3 to 6 % approximately), Candidatus Udaeobacter (~3 % -~6 %), Bryobacter (~3 % -~6 %), Puia (~1 % -~3 %), Acidothermus (~1 % -~2 %) and Streptococcus (~1 % -~2 %) (Fig. 3C, File S2). The bacterial genera profile in inside the ruts was deeply different than in between the ruts and in off-trail samples (Fig. 3C). In Inside the ruts samples, the genera Candidatus Udaeobacter, Candidatus Solibacter, Granulicella, and Bryobacter were more abundant than off-trail and between the ruts samples (Fig. ...

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... (6 % -20 %), Rozellomycota (<1 % -5 %), Zoopagomycota (<1 % -2 %) and Olpidiomycota only detected in between the ruts samples at T1 with a maximum relative abundance of 2.24 % (Fig. S3 B). At the order level, the orders Helotiales and Chaetothyriales showed a relative abundance higher than 3 % (range of ~18 % -~30 %) in all soil conditions (Fig. 3B, File S2). At T1 and T2 the relative abundances of some taxa significantly varied between levels of soil compaction reflecting the diversity in structure of fungal communities. Indeed, at T1 the order Tremellales was significantly less abundant in not-compacted than compacted soil samples (Fig. 3B, File S2), while Capnodiales and the order GS37 ...

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... % (range of ~18 % -~30 %) in all soil conditions (Fig. 3B, File S2). At T1 and T2 the relative abundances of some taxa significantly varied between levels of soil compaction reflecting the diversity in structure of fungal communities. Indeed, at T1 the order Tremellales was significantly less abundant in not-compacted than compacted soil samples (Fig. 3B, File S2), while Capnodiales and the order GS37 showed the opposite trend, being significantly enriched in not-compacted soil samples ( Fig. 3B and File S2). At T2, the order Trichosporonales were significantly less abundant in the compacted soil conditions (File ...

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... the fungal communities, thirty-one genera displayed an average relative abundance of 1 %, or higher, in at least one group (Fig. 3D, File S2). Among these, the genera characterized by a high relative abundance were Cenococcum, with a range of mean relative abundance equal to ~3 % -~12 %, Ciliophora (~2 % -6 %), Cryptosporiopsis (~0.8 % -~7 %), Hygrophorus (<1 % -~10 %), Inocybe (<1 % -~14 %), Lambertella (<1 % -~7 %), Oidiodendron (~1.1 % -~3 %) and Saitozyma (~2 % -~5 %) ...

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... T1, the enzyme activities associated with the bacterial structure were arylsulfatase, protease, and alkaline phosphatase activity (Fig. 6A, File S3). Consistently with enzyme activity results (Fig. 2), the regression analysis showed that the arylsulfatase activity was positively associated with changes in bacterial structure in the undisturbed soil (off-trail samples), while the protease and alkaline phosphatase activities progressively increased along with changes in bacterial structure in medium compacted soil (between ruts ...

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... the increment of acid phosphatase activity was connected to changes in bacterial community composition of compacted soils (inside ruts samples), while other activities (β-glucosidase, alkaline phosphatase, and arylsulfatase activity) were more associated with undisturbed soils (offtrail samples) (Fig. 6B), although they showed lower activity (Fig. ...