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Article

Effects of Ridge-Furrow Film Mulching Patterns on Soil Bacterial Diversity in a Continuous Potato Cropping System

by
Shujuan Jiao
1,
Yichen Kang
1,
Weina Zhang
1,
Yuhui Liu
2,
Hong Li
3,
Wenlin Li
1 and
Shuhao Qin
1,*
1
College of Horticulture, Gansu Agricultural University, Lanzhou 730070, China
2
State Key Laboratory of Aridland Crop Science, Agronomy College, Gansu Agricultural University, Lanzhou 730070, China
3
College of Science, Gansu Agricultural University, Lanzhou 730070, China
*
Author to whom correspondence should be addressed.
Agronomy 2025, 15(8), 1784; https://doi.org/10.3390/agronomy15081784
Submission received: 19 June 2025 / Revised: 22 July 2025 / Accepted: 23 July 2025 / Published: 24 July 2025
(This article belongs to the Special Issue Sustainable Potato Production: Emerging Strategies for Disease Prevention and Cultivation)
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Abstract

Soil bacteria drive biogeochemical cycles and influence disease suppression, playing pivotal roles in sustainable agriculture. Using Illumina MiSeq sequencing, we assessed how six ridge-furrow film mulching patterns affect soil bacterial diversity in a continuous potato system. The Shannon index showed significantly higher diversity in fully mulched treatments (T2–T3) versus controls (CK), suggesting mulching enhances microbial community richness. This result suggests that complete mulching combined with ridge planting (T2) may significantly enhance bacterial proliferation in soil. The bacterial communities were predominantly composed of Acidobacteria, Pseudomonadota, Bacteroidota, Chloroflexota, and Planctomycetota. Among these, Acidobacteria showed the highest abundance, with ridge planting patterns favoring greater Acidobacteria richness compared to furrow planting. In contrast, Pseudomonadota exhibited higher abundance under half-mulching conditions than under complete mulching. At class level, Acidobacteria and Proteobacteria emerged as the most abundant groups, with Proteobacteria constituting 22.6–35.7% of total microbial populations. Notably, Proteobacteria demonstrated particular dominance under the complete mulching with ridge planting pattern (T2). At the genus level, Subgroup_6_norank represented the most dominant taxon among the 439 identified bacterial genera, accounting for 14.0–20.2% of communities across all treatments, with half-mulching ridge planting (T4) showing the highest relative abundance. Our findings demonstrate that different ridge-furrow film mulching patterns significantly influence soil microbial diversity. While traditional non-mulched (CK) and mulched flat plots (T1) exhibited similar impacts on bacterial community structure, other treatments displayed distinct taxonomic profiles. Complete mulching patterns, particularly ridge planting (T2), appear most conducive to microbial development, suggesting their potential to enhance soil biogeochemical cycling in continuous cropping systems. These results provide valuable insights for optimizing mulching practices to improve soil health in agricultural ecosystems.

1. Introduction

Potato (Solanum tuberosum L.) serves as both a staple food and cash crop in northwest China, playing a pivotal role in the region’s agricultural development [1,2]. However, extensive continuous cropping has led to declining cultivation areas due to growth inhibition phenomena, including increased disease incidence, pest infestation, yield reduction, and quality deterioration [3,4]. These challenges have become major constraints for sustainable potato production.
Soil microbial communities, particularly bacteria and fungi, serve as the fundamental drivers of agroecosystem functioning through three key mechanisms: nutrient cycling (mediated by nitrogen-fixing rhizobia and phosphate-solubilizing bacteria, which transform inaccessible nutrients into plant-available forms) [5]; pathogen suppression through antibiotic production and niche competition [6]; and soil structure maintenance through glomalin secretion and hyphal networks [7]. In potato cultivation systems, specific microbial taxa such as Pseudomonas and Bacillus play a critical role in biocontrol by suppressing Verticillium wilt pathogens [8], while arbuscular mycorrhizal fungi significantly improve phosphorus uptake under nutrient-limited conditions [9]. These microbial functions are not only intrinsically linked to soil health but also directly regulate essential processes, including organic matter decomposition (primarily driven by cellulolytic Bacteroidetes and Actinobacteria) and carbon sequestration (associated with Methanotrophs abundance) [10,11]. Notably, microbial community dysbiosis—characterized by the decline of beneficial taxa (e.g., Acidobacteria subgroup GP1) and proliferation of pathogenic microbes (e.g., Fusarium oxysporum)—has been widely recognized as a hallmark of continuous cropping systems [12,13], underscoring the necessity of soil microbiome research for sustainable agriculture. Traditional culture methods can only characterize a limited fraction of soil bacteria [14]. Molecular techniques developed over the past two decades now enable detection of previously unculturable microbial species through analysis of unique 16S rDNA sequences. These 16S rDNA-based methods have become standard for environmental microbial community characterization. The Illumina MiSeq platform, as a second-generation high-throughput sequencing technology, offers exceptional analytical precision, sensitivity, and automation [15,16], making it widely adopted in both medical and microbial research [17,18]. Plastic film mulching has emerged as a simple yet effective water conservation technique [19,20,21]. Current research has primarily focused on optimizing farmland moisture conditions [22,23], with studies demonstrating that ridge mulching reduces soil water evaporation while enhancing rainfall infiltration and root zone moisture availability [24]. Full mulching significantly improves soil water retention, moisture utilization efficiency, and physicochemical properties, while promoting microbial proliferation [25,26,27,28]. Evidence from tobacco studies confirms that plastic mulching enhances rhizosphere soil enzyme activities including catalase, urease, and invertase [29,30,31]. While existing research has mainly investigated continuous cropping obstacles under non-mulched conditions, the effects of different ridge-furrow mulching patterns on soil quality in potato systems remain poorly understood. Based on the evidence from the above studies, we hypothesized that full mulch treatments would enhance soil bacterial diversity relative to non-mulched controls. To test this hypothesis, we employed Illumina MiSeq sequencing to characterize how different ridge-furrow mulching patterns influence soil bacterial communities in a continuous potato system and identify optimal mulching configurations for maintaining soil health through microbial community modulation. Our objectives were to: elucidate the impact of furrow-ridge mulching on soil quality in continuous potato fields and establish a theoretical foundation for mitigating potato continuous cropping obstacles.

2. Materials and Methods

2.1. Description of the Experimental Site

The field experiment was conducted from April–November 2022 at the Experiment Station of Dingxi Dry Farming Research Institute, Gansu Agricultural University, in collaboration with the Gansu Crop Genetic Improvement & Germplasm Innovation Key Laboratory. The site is located at an elevation of 1950 m in a typical semiarid rain-fed agricultural region with loessal soil of medium fertility [32]. The regional climate characteristics include mean annual temperature: 6.4 °C, annual cumulative temperature: 2239.1 °C, mean annual precipitation: 415.2 mm, annual evaporation: 1531 mm, and aridity index: 2.53. Basic soil physicochemical properties of the experimental site are presented in Table A1.

2.2. Experimental Design

This study employed a continuous cropping system, with 2022 representing the third consecutive year of potato cultivation at the experimental site. The experimental design consisted of six treatments (Table 1). arranged in a completely randomized block design with three replications (total 18 plots). “Xindaping”, a local main cultivar, was used for this study as the test cultivar. Standard agricultural black polyethylene (PE) plastic film (0.008 mm) was used. The sowing time was 30 April, and harvesting time was 1 October; The experimental layout was configured with the following spatial parameters: 2.0 m inter-block spacing; inter-plot spacing, 1.5 m between adjacent plots; plot dimensions: 6.6 m × 10 m (66 m2 per plot); The intra-row plant spacing was 35 cm, and inter-row spacing was 55 cm (mean value). Planting density was approximately 45,460 plants ha−1. The planting pattern is shown in Figure 1.

2.3. Soil Samples Collection

Rhizosphere soil samples were collected on 14 September 2022 (two weeks before harvest) from each plot following a standardized protocol to ensure consistency. Using an S-shaped sampling pattern, we carefully uprooted potato plants at six sampling points per plot to preserve the root-adhering soil. The roots were gently shaken to remove loosely attached bulk soil, and the remaining soil tightly bound to root surfaces (≤2 mm) was collected using ethanol-sterilized stainless steel tools as the rhizosphere fraction. For each plot, soil from all six sampling points was thoroughly homogenized to form a single composite sample, with three independent biological replicates per treatment (totaling 18 samples). Immediately after collection, visible roots and stones were removed, and subsamples were flash-frozen in liquid nitrogen before being transferred to −80 °C storage for microbial DNA extraction. The remaining soil was air-dried under natural conditions and sieved through a 2 mm mesh for physicochemical analysis.

2.4. Soil DNA Extraction

Genomic DNA was extracted from triplicate soil samples representing each of the six ridge-furrow film mulching treatments using the Power Soil DNA Isolation Kit (MO BIO Laboratories, Inc., Carlsbad, CA, USA) following the manufacturer’s protocol. DNA quality was assessed through 1% agarose gel electrophoresis with Gold View nucleic acid staining. To eliminate potential PCR inhibitors, particularly humic acids, DNA samples were subsequently purified using the GELASETM purification system (Epicenter, Madison, WI, USA) prior to downstream molecular analyses.

2.5. PCR Amplification and MiSeq Sequencing Analysis

PCR amplification of 16S rDNA genes was amplified using barcoded primers 515F (5′-GTGCCAGCMGCCGCGG-3′) and 907R (5′-CCGTCAATTCMTTTRAGTTT-3′) on an ABI GeneAmp® 9700 PCR system. Triplicate 20 µL reactions were performed for each treatment, containing: 4 µL 5× FastPfu Buffer, 2 µL 2.5 mM dNTPs, 0.4 µL FastPfu Polymerase, 0.8 µL of each primer (5 µM), and 10 ng template DNA, with ddH2O added to final volume. Thermal cycling conditions comprised: initial denaturation at 95 °C for 3 min; 27 cycles of 95 °C (30 s), 55 °C (30 s), and 72 °C (45 s); followed by final extension at 72 °C for 10 min. PCR products were verified by 2% agarose gel electrophoresis, purified using the AxyPrep DNA Gel Extraction Kit (Axygen, Union, CA, USA), and quantified after Tris-HCl elution. Equimolar amounts of purified amplicons were pooled, preserved with dry ice, and shipped to Majorbio Bio-Pharm Technology Co., Ltd. (Shanghai, China) for Illumina MiSeq paired-end sequencing (2 × 300 bp).

2.6. Statistical Analysis

Bioinformatic analysis of 16S rDNA sequences was performed following quality control procedures. First, paired-end reads from all six treatments were merged into single sequences based on overlap relationships. Operational Taxonomic Units (OTUs) were clustered at a 97% similarity threshold using UPARSE (v.11.0.667), with representative sequences selected for each OTU. Taxonomic assignment was conducted by BLASTn (v.2.15.0) analysis against the GenBank database. Phylogenetic relationships were reconstructed using neighbor-joining algorithms in MEGA 5.0 with 1000 bootstrap replicates. Alpha diversity metrics, including species richness (observed OTUs) and Shannon–Wiener index, were calculated in Mothur (v.1.35.1) after rarefaction to the minimum sequencing depth. Beta diversity analysis was performed using principal component analysis (PCA) implemented in CANOCO 4.5, with log-transformed OTU abundance data to examine community structure variations among different planting patterns.

3. Results

3.1. Soil Physicochemical Property

The soil physicochemical properties across six planting patterns are presented in Table 2. Comparative analysis revealed distinct treatment effects. Soil moisture content was significantly higher in ridge-planting systems compared to both flat plots (mulched and non-mulched). Soil pH showed an inverse relationship, with fully mulched treatments exhibiting more neutral pH values (7.3–7.5). The analysis revealed significant treatment effects on soil organic carbon and organic matter accumulation (p < 0.05), with the fully mulched treatments (T2 and T3) demonstrating substantially higher OC and OM content compared to other treatments. Notably, the ridge-based full mulching system (T2) showed 5.7% and 5.8% greater OC and OM accumulation, respectively, than the furrow-based full mulching (T3), likely reflecting enhanced organic matter preservation under ridge configurations. The half-mulched treatments exhibited intermediate values that were consistently greater than the non-mulched control (CK) but lower than fully mulched systems, establishing a clear treatment gradient of T2 > T3 > T1 > T4 > T5 > CK in terms of soil organic matter enrichment. Nutrient availability patterns differed markedly among treatments. Available nitrogen exhibited non-linear variation without clear treatment-dependent patterns, suggesting complex mineralization dynamics. Available potassium followed CK > T5 > T3 > T1 > T2 > T4 (3.3–32.7% reduction in mulched treatments versus control), while available phosphorus demonstrated an opposite trend (T3 > T5 > T2 > T1 > T4 > CK, with 55.3–56.4% increases in optimal treatments). These results collectively indicate that ridge-furrow mulching systems significantly modify soil physicochemical parameters, with particularly strong effects on water retention and phosphorus availability.

3.2. 16S rDNA Optimizing Sequence Statistics

Raw sequencing data from Illumina MiSeq were subjected to rigorous quality filtering. Sequences with low quality scores (<Q20) or short read lengths (<50 bp) were removed during preprocessing. After quality control and noise reduction, we obtained a total of 183,389 high-quality 16S rDNA sequences, representing 72,683,859 bp with an average length of 396.34 bp. The final optimized sequence counts for each treatment were as follows: CK (16,847 reads), T1 (20,291 reads), T2 (18,694 reads), T3 (9876 reads), T4 (12,423 reads), and T5 (17,601 reads). The complete distribution of optimized sequences across all samples is detailed in Table 3.

3.3. Species Richness and Diversity of Bacterial Community Altered by Six Planting Patterns

Comparative analysis of bacterial α-diversity revealed distinct patterns among the six mulching treatments (Table 4). Operational taxonomic units (OTUs) clustered at 97% similarity demonstrated adequate sampling coverage (90%), though additional diversity likely remains undetected. Notably, the Chao1 estimator indicated substantial variation in species richness, following the hierarchy: T3 > T2 > T1 > T5 > T4 > CK. Fully mulched furrow planting (T3) supported 13.8% greater richness than conventional flat cultivation (CK), with statistically significant divergence (p < 0.05, ANOVA). This trend suggests ridge-furrow systems, particularly complete mulching configurations, enhance the proliferation of dominant bacterial taxa relative to traditional non-mulched practices.
Analysis of microbial α-diversity by Shannon’s index showed that the bacterial community changed significantly (p < 0.05) with different treatments. The Nestle’s index and Shannon’s index were higher in the full-film cover system compared to the control than in the half-film cover system. This gradient revealed that complete mulching increased diversity by 22–29% compared to partial mulching systems, which may support specialized microbiota through improved moisture retention and thermal buffering; these findings particularly highlight how the integrated ridge-and-furrow structure with all-plastic mulching creates optimal microhabitats for a functionally diverse soil microbiota, which outperforms the traditional practice of flat-field mulching.

3.4. Analysis of Soil Bacterial Community Structure in Different Furrow-Ridge Mulching Planting Patterns

Taxonomic analysis revealed that six core bacterial groups dominated the soil communities across all treatments (Figure 2), including: Subgroup-6-norank, WD2101_soil_group_norank, Xanthomonadales_uncultured, Cytophagaceae_uncultured, Anaerolineaceae_uncultured, RB41_norank. Notably, the relative abundance of Acidobacteria Subgroup_6 exhibited treatment-dependent variation, with its relative abundance following the order: T4 > T1 > T2 > T3 > CK > T5, suggesting preferential enrichment in ridge-associated and mulching conditions. Minor taxa (each <2% relative abundance) included functional groups such as cellulose degraders (Chryseolinea, Anaerolineaceae_unclassified), nitrifiers (Nitrospira, Nitrosospira), and drought-tolerant species (Blastocatella, Bryobacter). Multivariate analysis delineated two primary clusters: conventional flat cultivation systems (CK, T1) and engineered ridge-furrow configurations (T2–T5). These findings demonstrate that ridge-furrow mulching significantly restructures soil microbial communities, with complete mulching promoting greater microbial abundance and diversity compared to partial or non-mulched systems.

3.5. Analysis of Soil Bacterial Community at the Phylum Under Different Furrow-Ridge Mulching Planting Patterns

Phylogenetic analysis of bacterial communities across six treatments revealed distinct compositional patterns at multiple taxonomic levels (Figure A1). The communities comprised 33 phyla, 73 classes, 163 orders, and 439 genera. Heatmap visualization demonstrated that ridge-furrow systems (T2–T5) formed a distinct cluster separate from conventional flat cultivation (CK, T1), with Acidobacteria and Proteobacteria showing opposing responses to mulching intensity and microtopography.
Taxonomic classification at the class level revealed six predominant bacterial groups: Acidobacteria (subgroups 1–6), Proteobacteria (α, β, γ, δ subclasses), Actinobacteria, Anaerolineae, Phycisphaerae, and Gemmatimonadetes. Acidobacteria emerged as the dominant class, followed by Proteobacteria, while other classes collectively represented 1.0–6.0% of communities. Acidobacteria abundance exhibited treatment-specific variation: T4 > T3 > T2 > CK > T5 > T1. Notably, the α-, β-, γ-, and δ-Proteobacteria subclasses demonstrated contrasting distribution patterns, with γ-Proteobacteria particularly enriched in T4 (32.6%) compared to other treatments. The fully mulched ridge system (T2) supported significantly greater bacterial richness (p < 0.05) across all dominant classes. Relatively, compared to six different furrow-ridge mulching patterns, the bacteria in completely mulched planting on the ridge are more abundant.
Microbial community analysis at the order level identified seven predominant taxonomic groups in continuous cropping soils: Acidimicrobiales, Anaerolineales, Burkholderiales, Cytophagales, Myxococcales, Planctomycetales, and Rhizobiales. Among these, Cytophagales emerged as the most abundant order, demonstrating significant treatment-dependent variation (p < 0.05). Notably, the fully mulched ridge system (T2) supported the highest relative abundance of Cytophagales. This pattern suggests that the combined effects of complete soil coverage and ridge microtopography create favorable conditions for cellulose-degrading bacteria, as Cytophagales are known to play important roles in plant polysaccharide decomposition. Secondary dominant orders showed distinct distribution patterns, with Burkholderiales and Rhizobiales being particularly enriched in treatments with partial mulch coverage, potentially indicating differential responses to soil moisture and temperature regimes among functional groups.
Continuous cropping soil bacteria contain 439 categories, such as Subgroup_6_norank, Blastocatella, Bryobacter, Chryseolinea, Cytophagaceae, Gemmatimonadaceae, Steroidobacter, and Xanthomonadales. Subgroup_6_norank in six treatments of soil vary from 14.0~20.2%, and others range between 1.0~7.0%.
Phylogenetic analysis of bacterial communities across six treatments revealed distinct compositional patterns at the phylum level (Figure 3). The dominant phyla included Pseudomonadota, Bacillota, Actinomycetota, Verrucomicrobiota, Planctomycetota, Elusimicrobiota, Armatimonadota, Chloroflexota, and Gemmatimonadota. Among these, Acidobacteriota (30.6%) and Pseudomonadota (25.5%) collectively represented over 50% of the total sequences across all samples, establishing them as the core microbial phyla in this community. Notably, Chloroflexota exhibited planting mode-dependent variations, with its highest abundance (6.2%) observed in the T3 group and consistently lower levels (5.3%) in the CK group, suggesting potential sensitivity to planting practices. In contrast, the T2 treatment showed a significant reduction in Actinomycetota abundance (2.8% lower than CK). Meanwhile, the T4 group demonstrated a marked increase in Cyanobacteria. A pronounced shift in the Pseudomonadota/Bacteroidota ratio was observed, with higher values (1.6–2.8) in the T3, T4, and T5 groups compared to the T1 group (0.45). This disparity may reflect divergent carbon source utilization strategies among the treatments.

3.6. PCA of Soil Bacterial Community in Different Furrow-Ridge Mulching Planting Patterns

PCA of OTU clusters (97% similarity threshold) revealed distinct separation patterns among the six planting systems (Figure 4). The first three principal components collectively explained 66.88% of total variance (PC1: 34.85%, PC2: 19.14%, PC3: 12.89%), demonstrating significant treatment-induced differentiation in bacterial community structure. The coordinated results from PCA analysis provide robust evidence that different furrow-ridge mulching planting patterns can change the soil quality and result in variation in soil bacterial profile. Furthermore, the results of clustering analysis also strengthen this viewpoint.

4. Discussion

Continuous cropping obstacles have gained increasing global attention in agricultural research [33]. Substantial evidence has demonstrated that multiple factors contribute to plant growth decline under continuous cropping systems, with soil microbial community dysbiosis being identified as a predominant driver of these obstacles [34,35]. Continuous cropping systems exhibit a significant reduction in beneficial soil bacteria and actinomycetes populations, while fungal abundance shows a contrasting increase [36]. As soil bacteria play essential roles in all aspects of nutrient cycling, this microbial community imbalance ultimately results in decreased soil fertility and impaired nutrient retention [14,37,38]. Film mulching enhances microbial communities primarily through three mechanisms: microclimate regulation, resource allocation, and physical protection. First, by stabilizing soil moisture and moderating temperature fluctuations, mulching creates favorable conditions for moisture-sensitive taxa such as Acidobacteria. Second, the increased root exudation observed under mulching provides abundant labile carbon sources that preferentially support copiotrophic microorganisms. Third, the physical barrier formed by the mulch reduces soil erosion and minimizes UV radiation exposure, thereby preserving critical microbial habitats. These mechanisms are consistent with the findings of Yang et al. [39], who reported similar enrichment of functionally important microbial groups in mulched agricultural systems. Our results provide further evidence that the plastic film mulching treatment can enhance the reproduction of microbial communities, highlighting its key role in improving the soil environment [40,41,42]. Mao et al. [43] specifically documented these improvements in saline-alkali soils, showing that plastic film mulching not only enhanced soil quality but also substantially modified rhizosphere microbial communities. Furthermore, Li et al. [44] established a direct correlation between soil microbial abundance, activity levels, and overall soil fertility. This study showed that the soil fertility of six different furrow-ridge mulching patterns was superior to that of traditional non-mulching flat plots; meanwhile, the results were consistent with the richness and diversity of soil bacteria, and completely mulching patterns were dominant, especially for completely mulching plants on the ridge pattern.
The application of Illumina MiSeq high-throughput sequencing technology enables more comprehensive characterization of soil bacterial community structure in potato continuous cropping systems compared to conventional molecular approaches [45,46,47,48]. Our analysis revealed that the predominant bacterial phyla in continuous cropping soils were Acidobacteriota and Pseudomonadota, with secondary representation from Bacteroidetes, Chloroflexota, and Planctomycetota. Our results align with the observations of Arunrat et al. [49], where Pseudomonadota and Acidobacteriota were enriched in structured agricultural soils (e.g., terraces), while Nitrospira dominated the rhizosphere. This consistency across studies suggests that soil structuring—whether through terracing or ridge-furrow mulching—selectively promotes these bacterial phyla due to improved microhabitat stratification and resource partitioning. Notably, the higher abundance of Nitrospira in our mulched treatments (T2/T3) further supports this shared pattern, likely driven by enhanced nitrogen cycling under modified soil physical conditions. Pseudomonadota, Chloroflexota, and Planctomycetota were also detected in nutrient-rich areas, such as charcoal and charcoal-mixed soil layers [50]. These findings align with the established research of Morigasaki [51] and Liu [52], confirming the characteristic predominance of oligotrophic Acidobacteria and metabolically versatile Pseudomonadota in agricultural soils under intensive cultivation. Microbial community analysis revealed significant variations in phylum-level distributions across different mulching systems. Acidobacteriota were significantly more abundant in ridge-mulching systems than in furrow-mulching treatments (p < 0.05), while Pseudomonadota showed an inverse pattern, with higher relative abundance in half-mulching than complete mulching systems. Flat-mulching plots (T1) uniquely harbored Chlamydiae (0.8 ± 0.1%), indicating microenvironment-specific selection pressures. At the class level, Acidobacteriota and Pseudomonadota dominated across all treatments. Genus-level profiling identified 439 taxonomic groups, with the uncultured Subgroup_6_norank being most prevalent (14.0–20.2%), particularly enriched in half-mulched ridge systems (20.2%). These results demonstrate that ridge-furrow configurations and mulching completeness collectively shape soil bacterial communities through modification of microhabitat conditions.
At the genus level, the enrichment of beneficial Pseudomonas and Bacillus under full mulching conditions mirrors findings from other mulched agricultural systems [28], suggesting these taxa may serve as universal indicators of improved soil health under plastic mulching regimes. Conversely, the significant reduction in Fusarium oxysporum aligns with observations in tomato continuous cropping systems [53], highlighting shared pathogen suppression mechanisms across solanaceous crops. Certain Pseudomonas species have been associated with plant pathogenesis [54], though it should be noted that many Pseudomonas strains are beneficial plant growth promoters [55]. Our data show changes in Pseudomonas abundance (Figure 3), but further strain-specific characterization would be needed to determine their functional roles. This finding supports established research demonstrating that high microbial diversity creates suppressive soils where bacterial wilt pathogens struggle to establish [56,57,58,59]. The observed microbial community restructuring under ridge-furrow mulching systems, particularly the enrichment of beneficial taxa and suppression of pathogens, may simultaneously enhance soil biogeochemical cycling while providing natural disease control, representing a potential dual benefit of these modified cultivation practices. Microbial-driven processes (e.g., nutrient cycling, pathogen suppression) primarily involve the conversion of soil nutrients that are poorly accessible to plants (e.g., minerals, organic matter) into bioavailable forms. This transformation generates highly efficient nutrient supplies, which fundamentally underpin crop yield and quality—though these effects may not be immediately observable [60,61]. In our study, the enrichment of microbial taxa such as Subgroup_6_norank and Blastocatella suggests a potential enhancement of nitrogen mineralization, which could indirectly promote plant growth. However, direct validation of this relationship would require comprehensive yield measurements in future field trials. In future work, multi-season field experiments with paired microbial/agronomic monitoring.
Although the 97% OTU clustering approach facilitated direct comparison with previous studies [62], as with most studies, there are some limitations that should be considered. For example, OTU-based analysis may lack sufficient resolution to detect strain-level variations that could be functionally important, as evidenced by recent mock community analyses [63]. However, it has also been shown that OTU and ASV yielded results with greater than 90% agreement at the genus level [64], suggesting that the OTU clustering approach is still reliable. We will use the ASV method in future studies.

5. Conclusions

Our results demonstrate that complete mulching systems (T2, T3) significantly enhanced microbial proliferation and reshaped the dominant bacterial community composition compared to conventional flat cultivation. The most notable response was observed in Acidobacteria, with its subgroup Subgroup_6_norank demonstrating significant enrichment (14.0–20.2% relative abundance) under full mulching conditions. This selective enrichment likely results from the modified microclimate created by plastic mulching, which maintains optimal soil moisture and stabilizes temperatures, creating favorable conditions for these symbiotic microorganisms. The proliferation of these functional taxa correlates with observed improvements in soil nutrient cycling, suggesting that plastic mulching enhances soil health through both physical modification and biological mechanisms. While our findings provide important insights into the effects of specific cropping patterns on soil microorganisms, it should be noted that these results were obtained under specific environmental conditions of our study area. Direct application of these findings to other environments would require further validation studies accounting for different climatic and geographical conditions.

Author Contributions

Conceptualization, H.L.; methodology, Y.K., H.L., and S.J.; software, W.L.; validation, W.L. and Y.K.; formal analysis, W.L.; investigation, W.Z.; resources, S.Q., Y.K., and Y.L.; data curation, S.J., W.L., and Y.K.; writing—original draft preparation, S.J.; writing—review and editing, W.L.; visualization, Y.L.; supervision, Y.L., H.L., and S.Q.; project administration, S.Q.; funding acquisition, S.Q. All authors have read and agreed to the published version of the manuscript.

Funding

This research was supported by the outstanding Graduate Student Innovation Star Project in Gansu Province, China (2025CXZX-799), the National Natural Science Foundation of China (32060441, 32260455, 32201810), the earmarked fund for Agriculture Research System of China (CARS-09-P14), the Key Research and Development Program of Gansu Province, China (25YFNA042).

Data Availability Statement

The datasets used and/or analyzed during the current study are available from the corresponding authors upon reasonable request.

Acknowledgments

We thank the teachers and staff of the School of Horticulture of Gansu Agricultural University and the State Key Laboratory of Crops in Arid Habitat of Gansu Province for their on-site and technical support. The authors are grateful for the funding support. We also thank the two anonymous reviewers for their useful comments on an early version of this manuscript.

Conflicts of Interest

The authors have declared that no conflict of interest exists.

Abbreviations

The following abbreviations are used in this manuscript:
ECElectrical conductivity
OCOrganic carbon
OMOrganic matter
SOCSoil organic carbon
TNTotal nitrogen
TPTotal phosphorus
TKTotal potassium
ANAvailable nitrogen
AKAvailable potassium
APAvailable phosphorus
PCAPrincipal component analysis
PCRPolymerase Chain Reaction

Appendix A

Table A1. Basic soil properties.
Table A1. Basic soil properties.
SchemepHSOC
(g/kg)		EC (μS/cm)SOM (g/kg)TNTPTK
(g/kg)(g/kg)(g/kg)
0–208.310.2632912.30.871.9213.26
Soil organic carbon (SOC), electrical conductivity (EC), soil organic matter (SOM), total nitrogen (TN), total phosphorus (TP), total potassium (TK).
Agronomy 15 01784 g0a1
Figure A1. The heatmap of soil bacterial genera in the different soils. A basis of the vertical and horizontal clustering of an OTU (species) is shown in the Figure. The similarities and differences in community composition from different samples under the classification level were reflected by color gradient and similar degree.
Figure A1. The heatmap of soil bacterial genera in the different soils. A basis of the vertical and horizontal clustering of an OTU (species) is shown in the Figure. The similarities and differences in community composition from different samples under the classification level were reflected by color gradient and similar degree.
Agronomy 15 01784 g0a1

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Agronomy 15 01784 g001
Figure 1. Planting patterns.
Figure 1. Planting patterns.
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Figure 2. Bacterial genera in the different soils.
Figure 2. Bacterial genera in the different soils.
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Figure 3. The relative abundances at the phylum level.
Figure 3. The relative abundances at the phylum level.
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Figure 4. Principal component analysis (PCA) of Q20 soil bacterial community in the different soils. (a) PC1 and PC2, (b) PC1 and PC3.
Figure 4. Principal component analysis (PCA) of Q20 soil bacterial community in the different soils. (a) PC1 and PC2, (b) PC1 and PC3.
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Table 1. Experimental treatment.
Table 1. Experimental treatment.
TreatmentOperation
CKNon-film mulched flat plot
T1Half-mulched flat plot (70 cm) alternated with strips of bare land (40 cm) without ridges. Potato planted in two rows in the mulched plot and spaced at 40 cm
T2Fully mulched ridge cropping (70 cm) alternated with narrow ridges (40 cm) and ridge cropping. Potato planted in two rows in the mulched ridges and spaced at 40 cm
T3Fully mulched furrow cropping (70 cm) with narrow ridges (40 cm). This was similar to T2 with the exception of planting at the bottom of the furrows (two furrows)
T4Half-mulched ridge cropping (70 cm) alternated with bare land (40 cm) that had no ridges and mulching. Potatoes were planted in two rows in the mulched ridges plots and spaced at 40 cm
T5Half-mulched furrow cropping (70 cm) alternated with bare land (40 cm) that had no ridges and mulching. Two rows of potatoes were planted in the non-mulched plots and spaced at 40 cm.
Table 2. Physicochemical properties of soil profile.
Table 2. Physicochemical properties of soil profile.
TreatmentMoisture/%pHEC (µs/cm)OC
(g·kg−1)		OM
(g·kg−1)		AN
(mg·kg−1)		AK
(mg·kg−1)		AP
(mg·kg−1)
CK7.3 ± 0.08 b7.8 ± 0.06 a284 ± 24.13 a12.3 ± 0.4 c21.2 ± 0.7 c22.1 ± 1.16 a200.7 ± 19.32 a18.7 ± 3.84 b
T18.9 ± 0.01 ab7.7 ± 0.05 ab276 ± 4.16 a14.1 ± 0.5 b24.3 ± 0.9 ab20.2 ± 1.91 ab124.7 ± 17.42 b25.8 ± 1.81 ab
T211.0 ± 0.22 a7.5 ± 0.17 ab232 ± 8.74 a16.8 ± 0.6 a29.0 ± 1.0 a16.9 ± 0.29 c135.0 ± 15.28 ab29.0 ± 4.21 a
T310.5 ± 0.16 a7.3 ± 0.03 b241 ± 8.50 a15.9 ± 0.5 a27.4 ± 0.9 a18.9 ± 0.27 bc193.3 ± 34.92 a29.2 ± 3.99 a
T410.3 ± 0.18 a7.5 ± 0.07 ab244 ± 87.09 a13.7 ± 0.4 b23.6 ± 0.7 b19.3 ± 0.41 b146.3 ± 24.33 ab18.0 ± 6.92 b
T510.6 ± 0.08 a7.6 ± 0.02 ab283 ± 22.72 a13.2 ± 0.4 b22.8 ± 0.7 bc20.6 ± 0.92 ab150.0 ± 26.41 ab18.9 ± 2.46 b
Data are means of three replicates with SE. The error bars represent the mean ± SE. Statistical significance of differences was determined by the ANOVA (p ≤ 0.05). Electrical conductivity (EC), organic carbon (OC), organic matter (OM), available nitrogen (AN), available potassium (AK), available phosphorus (AP). Different lower case letters denote significant difference at p ≤ 0.05.
Table 3. 16S rDNA optimizing sequence statistics.
Table 3. 16S rDNA optimizing sequence statistics.
Planting PatternsSequence Number
CK16,847
T120,291
T218,694
T39876
T412,423
T517,601
Table 4. Diversity and richness of soil bacterial community in different furrow-ridge mulching planting patterns.
Table 4. Diversity and richness of soil bacterial community in different furrow-ridge mulching planting patterns.
Planting PatternsOTU NumberChaoShannonCoverage/%
CK13841473 b5.97 d97.41
T113501624 ab6.05 bc97.22
T213441670 a6.14 a97.31
T311021677 a6.12 a94.48
T412061537 ab6.02 cd95.55
T513341613 ab6.09 ab97.41
Statistical significance of differences was determined by the ANOVA (p < 0.05). Different lower case letters represent significant differences at p < 0.05.
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Jiao, S.; Kang, Y.; Zhang, W.; Liu, Y.; Li, H.; Li, W.; Qin, S. Effects of Ridge-Furrow Film Mulching Patterns on Soil Bacterial Diversity in a Continuous Potato Cropping System. Agronomy 2025, 15, 1784. https://doi.org/10.3390/agronomy15081784

AMA Style

Jiao S, Kang Y, Zhang W, Liu Y, Li H, Li W, Qin S. Effects of Ridge-Furrow Film Mulching Patterns on Soil Bacterial Diversity in a Continuous Potato Cropping System. Agronomy. 2025; 15(8):1784. https://doi.org/10.3390/agronomy15081784

Chicago/Turabian Style

Jiao, Shujuan, Yichen Kang, Weina Zhang, Yuhui Liu, Hong Li, Wenlin Li, and Shuhao Qin. 2025. "Effects of Ridge-Furrow Film Mulching Patterns on Soil Bacterial Diversity in a Continuous Potato Cropping System" Agronomy 15, no. 8: 1784. https://doi.org/10.3390/agronomy15081784

APA Style

Jiao, S., Kang, Y., Zhang, W., Liu, Y., Li, H., Li, W., & Qin, S. (2025). Effects of Ridge-Furrow Film Mulching Patterns on Soil Bacterial Diversity in a Continuous Potato Cropping System. Agronomy, 15(8), 1784. https://doi.org/10.3390/agronomy15081784

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