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Article

Spatiotemporal Profiling of the Pathogen Complex Causing Common Bean Root Rot in China

1
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China
2
Department of Plant Pathology, China Agricultural University, Beijing 100193, China
3
Beijing Qigao Biologics Technology Co., Ltd., Beijing 100193, China
*
Authors to whom correspondence should be addressed.
Agriculture 2025, 15(13), 1426; https://doi.org/10.3390/agriculture15131426
Submission received: 18 May 2025 / Revised: 24 June 2025 / Accepted: 30 June 2025 / Published: 2 July 2025

Abstract

Root rot, a globally devastating disease of common bean (Phaseolus vulgaris L.), remains a major constraint on bean production across China. Despite its agricultural impact, the pathogen complex associated with this disease has been poorly characterized in most provinces. To address this critical knowledge gap, we conducted nationwide surveys during 2016–2018, systematically sampling 1–10 symptomatic plants from each of 121 (2016) and 170 (2018) field sites across 17 provinces in China’s major vegetable production regions. Isolates obtained from symptomatic root tissues underwent morphological screening, followed by molecular identification using partial sequences of EF1-α for Fusarium species and ITS regions for other genera. Pathogenicity of representative isolates was subsequently confirmed through controlled greenhouse assays. This integrated approach revealed fourteen fungal and oomycete genera, with Fusarium (predominantly F. oxysporum and F. solani) and Rhizoctonia (R. solani) emerging as the most prevalent pathogens. Notably, pathogen composition exhibited significant regional variation and underwent temporal shifts across developmental stages. Additionally, F. oxysporum, F. solani, and R. solani demonstrated significant interspecies associations with frequent co-occurrence in bean root rot systems. Collectively, this first comprehensive characterization of China’s common bean root rot complex not only clarifies spatial–temporal pathogen dynamics but also provides actionable insights for developing region- and growth stage-specific management strategies, particularly through targeted control of dominant pathogens during key infection windows.

1. Introduction

Common bean (Phaseolus vulgaris L.), including both dry bean and green bean varieties, represents the world’s third most economically important edible legume [1,2,3,4]. However, its global production faces significant challenges from root rot, a destructive soil-borne disease [5]. This disease manifests through several characteristic symptoms including seedling mortality, growth stunting, chlorotic foliage, and necrosis of root and vascular systems [6,7]. Depending on disease severity, yield losses can range from 25% to complete crop failure during severe epidemics [5].
The pathogens causing common bean root rot were highly diverse and complex [8,9]. Based on current reports, the major pathogens include fungi and also oomycetes. For pathogenic fungi, the genus Fusarium has the highest number of reported species, including the five pathogens F. solani, F. oxysporum, F. equisetii, F. lateritium, and F. reticulatum [6,10,11]. In addition to Fusarium, other fungal pathogens like Sclerotium spp. [11,12], Alternaria spp. [13], Rhizoctonia solani [10,12], Plectosphaerella cucumerina [14], Macrophomina phaseolina [11], and Colletotrichum spaethianum [15] also have been reported to cause root rot in common bean. Among the documented oomycete pathogens, Aphanomyces euteiches [16] and Pythium spp. are the most frequently reported [10,17,18]. Critically, these pathogens exhibit divergent fungicide sensitivities and epidemiological behaviors. Therefore, understanding the pathogen complex is essential for effective disease management.
Previous studies on the geographical distribution of root rot pathogens in common bean revealed distinct regional patterns (Table A1). Fusarium solani f. sp. phaseoli is the dominant pathogen in Latin America and Africa [19,20], and F. lateritium is the predominant species in Nuevo León, Mexico [21], while more complex multi-pathogen assemblages were observed in several regions: in Iran (F. solani, R. solani, F. oxysporum, Tiarosporella phaseolina, F. equisetii, P. ultimum and P. aphanidermatum [10]), Mexico (F. oxysporum, M. phaseolina, F. solani, F. lateritium and F. reticulatum [11]), and Puerto Rico (F. solani, M. phaseolina, S. rolfsii, P. aphanidermathum, P. graminicola, and R. solani [11]). These findings demonstrate significant variation in pathogen composition across different geographical locations.
China ranks among the world’s leading producers of common beans [22]. However, bean production faces significant threats from root rot disease, with reported yield losses reaching 84% in some regions and complete crop failure in severe cases [23,24]. Despite its economic impact, the pathogen complex causing common bean root rot in China remains poorly characterized. Current understanding relies on fragmented studies, primarily reporting the dominance of F. solani f. sp. phaseoli in localized areas [24,25,26]. Notably, no comprehensive study has systematically investigated the composition and geographical distribution of root rot pathogens across China’s diverse bean-growing regions. This critical knowledge gap continues to impede the development of region-specific, effective control strategies.
To address these critical knowledge gaps, this study was designed with three integrated research objectives: (I) to systematically collect and identify root rot pathogens from common bean across China’s major production regions, (II) to investigate the geographic distribution patterns of predominant pathogen groups, and (III) to comparatively analyze temporal variations in pathogen complex composition throughout different growth stages of common bean.

2. Materials and Methods

2.1. Sample Collection

Sampling was conducted across China’s primary vegetable production zones using randomized site selection. During field surveys conducted between 2016 and 2018, 291 symptomatic root samples were collected: 121 samples from 16 provinces in 2016, supplemented by 170 additional samples from 9 provinces in 2018 (Figure 1, Table A2 and Table A3). For each sampling site, the common bean growth stage, classified as seeding (V3–V4: first to third trifoliate leaf fully opened), pod-filing (R7-R8: pod formation to filing), or maturity (R9: physiological maturity) [27], as well as the geographic coordinates (latitude and longitude) and elevation (meters above sea level) were documented. Sampling protocols involved carefully excavating 1–10 symptomatic plants per site, gently removing soil from roots, immediately storing specimens in sterile kraft envelopes, transporting them on dry ice, and maintaining samples in a 4 °C refrigerator until pathogen isolation.

2.2. Isolation and Purification

Root samples were rinsed under tap water for approximately two minutes. For each root system, five to six 1 cm segments were excised from the lesion margins. These fragments were surface sterilized in 1% NaClO for two minutes, followed by three sterile distilled water rinses. After air-drying on sterilized filter paper, fragments were placed on amended potato dextrose agar (PDA, composition per liter: broth of 200 g pilled potato boiled, 20 g dextrose, 18 g agar, and 100 mg streptomycin sulfate and penicillin). Following incubation at 25 °C for 3–5 days, hyphal tips from colony edges were transferred to fresh PDA plates. Isolates were subsequently purified either through single-spore isolation (for sporulating cultures) or single hyphal tip culture (for non-sporulating cultures). For long-term storage, a culture plug from each purified colony was inoculated into 1 mL potato dextrose broth (PDB, PDA without agar) in a 2 mL centrifuge tube. The cultures were grown at 25 °C with 200 rpm shaking for 4–5 days before storage at 4 °C [28].

2.3. Morphological and Molecular Identification

For morphological characterization, each preserved isolate was subcultured by transferring a small mycelial plug onto both water agar (WA, composition per liter: 18 g agar) and potato sucrose agar (PSA, composition per liter: broth of 200 g pilled potato boiled, 20 g sucrose, 18 g agar) plates, followed by incubation at 25 °C for 3–7 days. Initial microscopic examination was performed using a microscope (AxioCam ERc 5s, Carl Zeiss Microscopy GmbH, Jena, Germany) to assess hyphal structures and sporulation morphology. Suspected Fusarium isolates underwent further characterization through culturing on carnation leaf agar (CLA) plates [29], maintained at 25 °C in dark for 7 days. These cultures were then examined under a Carl Zeiss microscope (AxioCam ERc 5s, Carl Zeiss Microscopy GmbH, Jena, Germany) to observe key diagnostic features including conidial morphology, sporulation structures, and chlamydospore formation. Final morphological identification was conducted following established taxonomic criteria from three authoritative sources: The Fungal Identification Manual, The Fusarium Laboratory Manual, and Chinese Fungal Chronicles [29,30,31].
For further molecular identification, at least two representative isolates per morphologically identified species were randomly selected and cultured on PDA plates until colonies covered two-thirds of the surface. After, mycelia were harvested for DNA extraction using a plant genomic DNA Kit (Tsingke BioTechnology Co., Ltd., Beijing, China) with subsequent storage at −20 °C. The EF-1α gene of Fusarium isolates was amplified using primers EF1/EF2 [32], and the ITS region of other isolates was amplified using universal primers ITS1/ITS4 [33]. The resulting amplicons were sequenced by Sangon Biotech Co., Ltd. (Shanghai, China) and analyzed via NCBI BLAST (version 2.8.1, URL: http://www.ncbi.nlm.nih.gov, accessed on 28 December 2018).

2.4. Pathogenicity Test

To satisfy Koch’s postulate [34], pathogenicity tests were conducted on common bean seeds in greenhouse conditions using 80 representative isolates (minimum 2 per genus/species). To prepare inoculum, intact kernels of uniform size were soaked in double-distilled water for 12 h, sterilized at 121 °C for 30 min, and oven-dried at 80 °C for 2 h [35]. For each isolate, 20 prepared kernels were placed around a mycelial plug on PDA plates, then incubated at 25 °C for 7 days until complete mycelial colonization. The fully colonized wheat kernels were then used as inoculum for subsequent pathogenicity testing.
Common bean seeds (cv. English Red) were surface sterilized in 1% NaClO for two minutes, rinsed three times with sterile distilled water, and soaked in sterile water for 40 min. The sterilized seeds were then sowed in pots (8 cm × 15 cm) containing sterilized soil matrix. For inoculation, one mycelium-colonized wheat kernel was placed adjacent to each seed (one seed per pot, with five pots per replicate and three replicates per isolate), then covered with approximately 3 cm of sterilized soil. Control treatments received sterilized wheat kernels without mycelia. Plants were then maintained in a greenhouse at 25 °C with a 12 h photoperiod for 3–4 weeks before disease evaluation. Root rot severity was assessed using a 0–5 scale [36]: 0 (no symptoms), 1 (light discoloration or 1–10% root surface affected), 2 (heavy discoloration or 11–25% root surface affected), 3 (26–50% root surface affected), 4 (51–75% root surface affected), and 5 (76–100% root surface affected or plant death).

2.5. Data Analyses

Statistical analysis was performed using R software (version 3.6.2). Non-parametric analyses were conducted to evaluate differences in disease severity ratings across species. The Kruskal–Wallis test was first performed as an omnibus test to evaluate overall group differences using the kruskal.test function. Following a significant result (p < 0.05), Dunn’s post hoc test with Bonferroni adjustment was conducted for pairwise comparisons between species via the dunn.test function from the dunn.test package (v1.3.5).
To examine geographical and temporal variations in pathogen composition, we categorized isolates into four groups: Fusarium, Rhizoctonia, Alternaria, and other minor genera (including 11 additional fungal and oomycete genera). Sampling locations were classified into three geographical regions (North China, East China, and Southwest China; Figure 1 and Table A2). Detection frequency (F) of each pathogen group was calculated as: F = (number of group isolates in region/total region isolates) × 100 [37]. Frequency distribution data were transformed using centered log-ratio (CLR) normalization to address compositional constraints. This was implemented via the transform CLR. function from the RVAideMemoire package (v0.9-83). Overall group differences in CLR-transformed compositional profiles were assessed using permutational multivariate analysis of variance (PERMANOVA) with 999 Monte Carlo permutations. Analysis was conducted using the adonis2.function from the vegan package (v2.6-6) with Bray–Curtis dissimilarities. For significant PERMANOVA results (α = 0.05), univariate group differences in individual components were examined using non-parametric Kruskal–Wallis tests via kruskal.test (stats v4.3.1) and false discovery rate (FDR) adjustment of p-values using the Benjamini–Hochberg method via p.adjust(method = “fdr”).
Given that over 80% of samples yielded multiple major pathogens, potential associations among co-existing species were investigated. For key pathogen pairs (F. oxysporum-F. solani [Fo-Fs], F. oxysporum-R. solani [Fo-Rs], and F. solani-R. solani [Fs-Rs]), chi-square tests of independence (chisq.test) were conducted with the following interpretation framework: p ≥ 0.05: no significant association (accept null hypothesis of independence); p < 0.05 with observed co-occurrence > expected: positive association; p < 0.05 with observed co-occurrence < expected: negative association (antagonism). This analysis revealed whether pathogen pairs tended to co-colonize (positive association), compete (negative association), or occur independently in infected samples.

3. Results

3.1. Identification of Isolates from Common Bean Root Rot Samples

A total of 2299 isolates were obtained from 291 common bean root samples collected between 2016 and 2018. These isolates were identified through combined morphological and molecular characterization as belonging to fourteen genera, namely, Fusarium, Rhizoctonia, Alternaria, Penicillium, Pythium, Plectosphaerella, Chaetomium, Trichoderma, Ascodesmis, Aspergillus, Phytophthora, Colletotrichum, Botryotrichum, and Humicola. The molecular identification details are provided in Table A4. Fusarium was found to be the most predominant genus (59.0% of total isolates), followed by Rhizoctonia (16.1%) and Alternaria (9.1%). The remaining eleven genera were each represented at significantly lower frequencies (≤2.9%), as demonstrated in Figure 2.

3.2. Pathogenicity and Virulence Test

The results of pathogenicity test demonstrated that all 80 tested isolates could cause symptoms on inoculated common bean plants, including seedling death, plant stunting, leaf discoloration, and root rot, which were consistent with symptoms observed under natural field conditions. The original isolates inoculated were re-isolated from these infected plant tissues, while no symptoms were observed in mock-treated plants (Figure 3, Table A5). Significant variation in virulence was detected among pathogens (p < 0.05; Figure 4). R. solani isolates caused extremely higher disease severity than any other pathogens tested (p < 0.01). Phytophthora showed significantly higher virulence than Plectosphaerella spp. and Colletotrichum spp. (p < 0.05). There was no significant difference in virulence among the remaining pathogens tested.

3.3. Geographical Distribution of Major Pathogen Groups

PERMANOVA analysis revealed significant regional variations in pathogen composition across China’s three major production regions [Pr(>F) = 0.002; Table 1]. Fusarium was the dominant genus in all regions, though its isolation frequency showed substantial geographical variation, ranging from 48.6% in North China to 75.9% in Southwest China. In contrast, the isolation frequency distribution of Rhizoctonia across regions falls within a relatively narrow range from 14.8% in Southwest China to 19.6% in East China. The combined other-genera category was most frequent in North China (23.8%) and least common in Southwest China (5.9%). Notably, the frequency distribution of Alternaria, ranging from 3.3% in Southwest China to 12.2% in North China, exhibited significant variation across geographical regions (p < 0.05), whereas no other pathogens showed significant differences in their regional distribution patterns (Figure 5).

3.4. Regional Variation of Fusarium Species

Four Fusarium species (F. solani, F. oxysporum, F. virguliforme, and F. chlamydosporum) were isolated from all common bean root rot samples. Among these, F. oxysporum (50.9%) and F. solani (44.2%) collectively accounted for 95.1% of all Fusarium isolates, while F. virguliforme and F. chlamydosporum occurred at significantly lower frequencies. Correspondingly, isolation frequencies of the four Fusarium species showed significant variation (χ2 > 100, p < 0.0001; Table 2). The regional analysis revealed both dominant species F. oxysporum and F. solani showed even distribution across regions, however, F. chlamydosporum showed restricted distribution, only detected in North China (Table 2).

3.5. Growth Stage-Specific Variation of Major Pathogen Groups

PERMANOVA analysis revealed significant variations in pathogen group frequencies across three growth stages of common bean [Pr(>F) = 0.006; Table 3]. Fusarium maintained dominance at all stages, with its incidence progressively increasing from seedling (46.8%) to maturity (70.0%). Similarly, Rhizoctonia exhibited steady growth stage-dependent increases, initially ranked lowest among groups during seedling (11.7%) and pod-filling (15.0%) stages, but ultimately exceeding both Alternaria and other genera at maturity. On the contrary, the frequency of the combined “other genera” category gradually declined from 26.2% to 9.1%, as the common bean developed. However, only the frequency distribution of Alternaria exhibited significant variations across different growth stages (p < 0.05; Figure 6).

3.6. Growth Stage Variations in Fusarium Species Composition

F. solani and F. oxysporum collectively accounted for over 95% of Fusarium isolates across all growth stages, demonstrating clear dominance. Correspondingly, isolation frequencies of the four Fusarium species showed significant variation in each growth stage (p < 0.0001; Table 4). However, the frequencies of each Fusarium species showed no significant differences across different growth stages (p > 0.05; Table 4).

3.7. Pathogen Co-Occurrence in Bean Root Rot

Statistical analysis revealed significant species associations among F. oxysporum, R. solani, and F. solani (p < 0.001; Table 5). The observed co-occurrence patterns consistently exceeded theoretical expectations under random distribution assumptions. The observed co-occurrence of F. oxysporumF. solani, F. oxysporumR. solani, and F. solaniR. solani significantly exceeded theoretical random distribution values. In contrast, isolated occurrences of any single pathogen consistently fell below expected values. Simultaneous absence of pathogen pairs also occurred more frequently than predicted. These consistent patterns across all tested species combinations suggest strong ecological interactions among these dominant root rot pathogens.

4. Discussion

This study presented the first systematic characterization of the pathogen composition associated with common bean root rot across major production regions in China, addressing critical knowledge gaps in geographical distribution and growth-stage dynamics. Our investigation revealed a diverse 14-genus pathogen complex dominated by Fusarium (59.0%) and Rhizoctonia (16.1%), with nine novel pathogen identifications expanding global understanding of legume root rot etiology. Among them, F. virguliforme and F. chlamydosporum were newly confirmed as causal agents alongside seven previously unreported genera, namely, Chaetomium, Trichoderma, Ascodesmis, Aspergillus, Phytophthora, Botryotrichum, and Humicola [5,12,13,38]. Notably, R. solani exhibited significantly higher virulence than any other pathogens. These findings significantly broaden the known taxonomic diversity of bean root rot pathogens beyond existing worldwide reports, providing crucial insights for disease management strategies.
Significant regional variations in major pathogen group frequencies were observed across China’s three production regions, reflecting distinct environmental influences. Fusarium was dominant in all regions without significant variations, and the most virulent Rhizoctonia exhibited similar isolation frequency distribution in all regions too, while Alternaria showed significant variation across geographical regions. These patterns aligned with global reports of pathogen heterogeneity, such as F. solani f. sp. phaseoli dominance in Latin America and Africa [19,20,39,40] or F. lateritium in Mexico [21]. Regional climate, soil properties, and agronomic practices collectively shaped pathogen communities [41,42,43], underscoring the need for region-specific management strategies. At the species level, F. oxysporum and F. solani collectively accounted for > 95% of Fusarium isolates but displayed contrasting distributions. F. oxysporum dominated East and Southwest China, whereas F. solani prevailed in North China. Such divergence might reflect adaptations to local soil conditions or host genotypes, as reported previously [11,40,44].
Significant shifts in pathogen composition were observed across common bean growth stages, revealing distinct temporal dynamics. Fusarium prevalence progressively increased from seedling to maturity, consistent with its role as a primary root rot pathogen [10,11,45,46]. Species-level analysis revealed F. oxysporum dominated early stages (seedling to pod-filling), aligning with its association with seedling diseases [6,47,48], while F. solani increased at maturity, likely due to cumulative colonization [28]. Concurrently, Rhizoctonia frequencies exhibited growth stage-dependent escalation, surpassing other genera by maturity, potentially reflecting declining host resistance or pathogen accumulation [49]. Although these temporal patterns were derived from national-scale data, regional-scale investigations of developmental stage-associated pathogen community dynamics would enhance the development of targeted management strategies. These temporal dynamics underscore the necessity for stage-specific management strategies targeting dominant pathogens during critical developmental phases.
A key finding was that any two of the three species F. solani, F. oxysporum, and R. solani showed significant positive associations and frequently co-occurred in diseased roots, though all three rarely appeared simultaneously. This synergy might result from either shared environmental preferences (e.g., high nitrogen availability or specific soil microbiome conditions) or facilitated infection, whereby initial colonization by one pathogen modifies root physiology to benefit subsequent infections [38,43]. Further research is needed to elucidate the underlying mechanisms of these interactions and develop integrated control strategies targeting these co-occurring pathogens.
Our study establishes a scientific foundation for targeted root rot management in China. The documented regional variations and growth-stage dynamics of pathogens support the need for location-specific strategies, such as deploying resistant cultivars tailored to dominant local species or timing control measures to critical growth phases [9,50]. Additionally, the frequent co-occurrence of major pathogens (F. solani, F. oxysporum, and R. solani) underscored the necessity of integrated management measures, such as soil amendments to disrupt pathogen synergies, rather than single-species targeting [50,51].
To enhance root rot management, further work should characterize the virulence diversity within key pathogenic species, investigate soil microbiome-mediated modulation of pathogen interactions, and validate region-specific and growth stage-adapted control strategies through field trials. Such efforts will provide the scientific basis for developing sustainable, precision-based management approaches against this economically devastating disease.

5. Conclusions

Nationwide surveys identified Fusarium (F. oxysporum, F. solani) and Rhizoctonia (R. solani) as the dominant pathogens causing common bean root rot in China, with 14 fungal and oomycete genera detected. Pathogen composition varied regionally and across growth stages, with these key species showing high virulence and frequent co-infection. This first comprehensive study provides critical insights for developing targeted, region- and growth stage-specific control strategies to manage this devastating disease.

Author Contributions

L.Y.: conceptualization, investigation, methodology, formal analysis, data curation, and writing—original draft. X.-H.L.: conceptualization, methodology, supervision, and writing—review and editing. B.-M.W.: investigation, supervision, and writing—review and editing. Z.-M.Z.: investigation. S.-D.L.: conceptualization and resources. All authors have read and agreed to the published version of the manuscript.

Funding

This study was financially supported by the China Agricultural Research System (CARS-23-C04) and the Special Project of Ministry of Agriculture (201503112-2).

Institutional Review Board Statement

Not applicable.

Data Availability Statement

All data generated or analyzed during this study are included in this published article. The datasets used during the current study are available from the corresponding author on reasonable request.

Acknowledgments

The authors highly appreciate the assistance of Lu Xiao in the sampling activity.

Conflicts of Interest

Author Zeng-Ming Zhong was employed by the company Beijing Qigao Biologics Technology Co., Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Appendix A

Table A1. Worldwide pathogens causing root rot in common beans and their geographic distribution.
Table A1. Worldwide pathogens causing root rot in common beans and their geographic distribution.
Fungal Pathogen (Genus/Species)Main Legume Crop HostsPrimary SymptomsGeographic DistributionKey References
F. solaniP. vulgaris, Glycine max, Vigna unguiculataFoot and root rot, vascular wiltingLatin America, North America (Puerto Rico), Africa, Asia (Iran)[6,10,11]
Fusarium solani f. sp. phaseoliP. vulgarisRoot rot, vascular wiltingAmericas (United States, Puerto Rico, Spain), Africa, Asia[19,20]
F. oxysporumG. max, P. vulgaris, V. unguiculataRoot rot, vascular browningLatin America, Africa, Asia (Iran)[10]
F. equisetiiG. max, P. vulgarisFoot and root rotAsia (Iran)[10]
F. lateritiumP. vulgaris, Pisum sativumRoot rotNorth America (Mexico), Asia[19]
F. reticulatumG. max, P. vulgarisRoot rotNorth America (Puerto Rico), Asia[11]
Sclerotium spp.G. max, P. vulgaris, V. unguiculata Foot and root rotAfrica (Uganda)[11,12]
Alternaria spp.P. vulgarisRoot rotAfrica (Tunisia)[13]
Rhizoctonia solaniG. max, P. vulgaris, P. sativumRoot rot, damping-offNorth America (Puerto Rico), Asia (Iran), Africa (Uganda)[10,12]
Plectosphaerella cucumerinaP. vulgarisRoot rotAsia (China)[14]
Macrophomina phaseolinaP. vulgarisDry root rotNorth America (Puerto Rico), Asia [11]
Colletotrichum spaethianumP. vulgarisRoot rotAsia (China)[15]
Aphanomyces euteichesP. sativum, P. vulgarisRoot rotAmericas (United States), Europe[16]
Pythium spp.P. vulgaris, V. unguiculata, P. sativumFoot and root rotSouth America (Spain), Europe, Asia (Iran)[11,17,18]
Tiarosporella phaseolinaP. vulgaris, Medicago sativaDry wilt and foot rotAsia (Iran), Americas[10]
Table A2. Common bean root rot samples collected from each province in China during this study.
Table A2. Common bean root rot samples collected from each province in China during this study.
RegionProvinceCity/CountyVillageStageNumber of SamplesRoots per SiteTotal Roots
District
North ChinaShanxi51416R7–R9305150
Neimenggu112V3–V42510
Henan8924V3–V4; R7–R9302–5128
Hebei101629V3–V4; R7–R9451–8182
Beijing7744V3–V4; R7–R9583–6200
East ChinaZhejiang125V3–V4; R7–R874–1847
Shandong51313R9131081
Jiangxi111R9155
Anhui111R7–R811010
Southwest ChinaSichuan31021R93610360
Chongqing3514R92110210
Guangxi112R7–R92714
Guizhou115V3–V4; R7–R8122–635
Hunan3812V3–V4124–1077
-Liaoning345R99545
-Hainan115V3–V4; R7–R9113–753
-Jilin111R9155
-Total5595200-291-1612
Notes: V = vegetative; R = reproductive; V3–V4: from the first trifoliate leaf fully opened to the third trifoliate opened; R7–R8: from pod formation to pod-filing; R9: physiological maturity.
Table A3. The latitude and longitude information of the sampling sites.
Table A3. The latitude and longitude information of the sampling sites.
Sample No.Sampling ProvinceLatitudeLongitude
1Shanxi39.31020 113.18628
2Shanxi39.41713 113.17947
3Shanxi39.40783 113.15060
4Shanxi38.39267 111.94016
5Shanxi38.80105 111.66961
6Shanxi38.76749 111.61128
7Shanxi38.39197 111.86366
8Shanxi38.28847 111.97351
9Shanxi38.35871 111.93238
10Shanxi38.38697 111.87569
11Shanxi38.40367 111.87566
12Shanxi38.21078 111.93782
13Shanxi38.10853 111.99765
14Shanxi37.84183 113.52763
15Shanxi37.85327 113.61473
16Shanxi37.85450 113.63106
17Shanxi37.93801 113.60275
18Shanxi37.80781 113.62741
19Shanxi35.47936 112.63339
20Shanxi35.47436 112.61046
21Shanxi35.40627 112.47092
22Shanxi35.48027 112.49632
23Shanxi35.47671 112.52754
24Shanxi37.65262 112.70026
25Shanxi36.98486 111.89460
26Shanxi37.92152 113.62477
27Shanxi36.93915 111.90385
28Shanxi37.62325 112.52692
29Shanxi37.62237 112.54632
30Shanxi37.61422 112.46373
31Neimenggu40.93680 113.20556
32Neimenggu40.60025 115.63154
33Henan35.24107 113.52763
34Henan31.70716 115.13929
35Henan36.06631 114.32032
36Henan36.06601 114.32383
37Henan36.06714 114.32445
38Henan36.06712 114.32716
39Henan36.06746 114.32547
40Henan36.06569 114.32688
41Henan36.06366 114.32721
42Henan36.06117 114.33316
43Henan36.05825 114.33391
44Henan36.04255 114.34251
45Henan35.28853 114.02144
46Henan35.25991 114.03001
47Henan35.24929 114.04027
48Henan35.23720 114.03345
49Henan34.76818 113.24252
50Henan34.79415 113.20285
51Henan35.67050 114.29795
52Henan35.72525 114.34031
53Henan35.69275 114.36139
54Henan35.65642 114.35042
55Henan35.66290 114.40715
56Henan35.72135 114.42521
57Henan35.75791 114.41259
58Henan35.68262 114.41416
59Henan33.27565 111.56088
60Henan34.09981 113.85962
61Henan34.11798 113.70382
62Henan36.15724 114.41963
63Hebei40.51939 115.60522
64Hebei39.81131 116.81117
65Hebei39.59364 116.54935
66Hebei39.59364 116.54935
67Hebei39.82800 115.40550
68Hebei39.82800 115.40560
69Hebei39.82000 115.40190
70Hebei39.82000 115.40190
71Hebei39.82000 115.40190
72Hebei39.82800 115.40550
73Hebei39.82985 115.40976
74Hebei39.04724 115.65895
75Hebei39.04666 115.66029
76Hebei39.04436 115.65932
77Hebei38.43967 114.97497
78Hebei38.41491 114.96345
79Hebei38.34450 114.96623
80Hebei38.34595 114.96477
81Hebei36.33047 114.96307
82Hebei38.32317 114.96325
83Hebei38.32317 114.96325
84Hebei38.32317 114.96325
85Hebei37.46296 114.66158
86Hebei37.46664 114.67618
87Hebei37.46661 114.67642
88Hebei37.52309 115.57732
89Hebei37.51004 115.57800
90Hebei37.50952 115.58409
91Hebei37.51305 115.59563
92Hebei37.51468 115.59662
93Hebei37.51740 115.60588
94Hebei38.45041 115.79375
95Hebei38.40530 115.80514
96Hebei38.42277 115.75369
97Hebei38.42602 115.75132
98Hebei38.42508 115.75149
99Hebei38.45985 115.76659
100Hebei38.45985 115.76659
101Hebei39.46110 116.20728
102Hebei39.46115 116.20642
103Hebei39.46064 116.20638
104Hebei39.46064 116.20638
105Hebei39.45913 116.20924
106Hebei39.45918 116.21256
107Hebei39.45918 116.21256
108Beijing 40.13645 116.17446
109Beijing 40.10151 116.22118
110Beijing 40.02956 116.28713
111Beijing 40.02956 116.28713
112Beijing 40.03739 116.27167
113Beijing 40.04535 116.30024
114Beijing 40.02956 116.28713
115Beijing 40.02956 116.28713
116Beijing 40.02956 116.28713
117Beijing 40.02956 116.28713
118Beijing 40.02956 116.28713
119Beijing 40.02956 116.28713
120Beijing 40.10151 116.22118
121Beijing 40.10151 116.22118
122Beijing 40.02884 116.29159
123Beijing 40.02884 116.29159
124Beijing 40.02884 116.29159
125Beijing 40.02884 116.29159
126Beijing 40.02884 116.29159
127Beijing 40.15390 116.58520
128Beijing 40.20290 116.58460
129Beijing 40.22170 116.57330
130Beijing 40.21584 116.60732
131Beijing 40.24363 116.65337
132Beijing 40.24527 116.63536
133Beijing 40.27652 116.63219
134Beijing 40.28004 116.62683
135Beijing 40.36181 116.76081
136Beijing 40.22410 116.46140
137Beijing 40.22540 116.54210
138Beijing 40.22200 116.57140
139Beijing 39.66237 116.53007
140Beijing 39.65502 116.35959
141Beijing 39.63912 116.47253
142Beijing 39.64164 116.43046
143Beijing 39.61289 116.49752
144Beijing 39.66664 116.52510
145Beijing 39.65841 116.53228
146Beijing 39.62822 116.56745
147Beijing 39.61253 116.41731
148Beijing 40.11570 117.03000
149Beijing 40.10250 116.58490
150Beijing 40.91300 116.59100
151Beijing 40.43300 116.59210
152Beijing 40.45300 116.52350
153Beijing 40.35100 116.51400
154Beijing 40.34500 116.90450
155Beijing 40.06725 116.77033
156Beijing 40.07140 116.75619
157Beijing 39.61300 116.41226
158Beijing 39.57070 116.35904
159Beijing 39.55734 116.44816
160Beijing 39.53778 116.32705
161Beijing 39.59258 116.33069
162Beijing 39.66014 116.54935
163Beijing 39.57573 116.49532
164Beijing 39.59324 116.33205
165Beijing 39.56887 116.49686
166Zhejiang30.18900 120.26564
167Zhejiang30.24832 120.06353
168Zhejiang30.25751 119.97864
169Zhejiang30.28404 120.13571
170Zhejiang30.25176 120.07564
171Zhejiang30.25176 120.07564
172Zhejiang30.41065 119.83784
173Shandong36.60327 118.68391
174Shandong36.95311 118.82176
175Shandong35.48044 117.72460
176Shandong35.46022 117.68366
177Shandong35.58332 117.64191
178Shandong35.73337 117.91193
179Shandong35.61296 117.65528
180Shandong35.46193 117.74347
181Shandong35.97355 117.89570
182Shandong35.95890 117.90614
183Shandong35.15014 114.93485
184Shandong35.16870 114.75342
185Shandong35.14053 114.98570
186Jiangxi28.99013 116.81879
187Anhui30.63887 116.58448
188Sichuan31.12368 104.21607
189Sichuan31.11764 104.21550
190Sichuan31.11689 104.21462
191Sichuan31.11663 104.21406
192Sichuan31.11382 104.21097
193Sichuan31.11443 104.20951
194Sichuan31.01974 104.17745
195Sichuan31.09836 104.27236
196Sichuan31.10045 104.27108
197Sichuan31.10554 104.27395
198Sichuan31.10802 104.26935
199Sichuan31.11207 104.26560
200Sichuan31.11570 104.26127
201Sichuan31.11973 104.26169
202Sichuan31.12024 104.25909
203Sichuan31.12310 104.25439
204Sichuan31.12724 104.24940
205Sichuan31.12561 104.24385
206Sichuan31.02630 104.16052
207Sichuan31.01499 104.16545
208Sichuan31.01655 104.16694
209Sichuan31.01916 104.17009
210Sichuan31.01683 104.12060
211Sichuan31.01857 104.12406
212Sichuan31.01778 104.12712
213Sichuan31.01756 104.12735
214Sichuan31.01533 104.13420
215Sichuan31.01524 104.13410
216Sichuan31.01530 104.13946
217Sichuan31.01916 104.17010
218Sichuan31.01524 104.13410
219Sichuan31.12992 104.23796
220Sichuan31.00541 104.15568
221Sichuan31.02283 104.10629
222Sichuan31.01914 104.10843
223Sichuan31.10351 104.20873
224Chongqing30.77011 108.43954
225Chongqing30.00016 106.12687
226Chongqing29.99863 106.12575
227Chongqing29.99859 106.13578
228Chongqing29.99780 106.14043
229Chongqing29.99556 106.14550
230Chongqing29.99355 106.15266
231Chongqing29.99187 106.16047
232Chongqing29.98706 106.16505
233Chongqing29.98706 106.16506
234Chongqing29.78785 106.28156
235Chongqing29.79203 106.28292
236Chongqing29.79315 106.28454
237Chongqing29.79113 106.27917
238Chongqing29.79970 106.28218
239Chongqing29.80803 106.28344
240Chongqing29.80283 106.28382
241Chongqing29.80220 106.28448
242Chongqing29.80153 106.28524
243Chongqing29.80177 106.28537
244Chongqing29.98706 106.16505
245Guangxi23.32623 106.61212
246Guangxi23.32899 106.62158
247Guizhou 27.21687 107.57111
248Guizhou 27.21687 107.57111
249Guizhou 27.21687 107.57111
250Guizhou 27.21687 107.57111
251Guizhou 27.22345 107.55065
252Guizhou 27.21676 107.57098
253Guizhou 27.20031 107.54578
254Guizhou 27.20154 107.54572
255Guizhou 27.18461 107.58902
256Guizhou 27.20154 107.54572
257Guizhou 27.20154 107.54572
258Guizhou 27.22196 107.55738
259Hunan27.28936 110.60734
260Hunan25.54479 113.65578
261Hunan25.54967 113.60865
262Hunan25.55936 113.69013
263Hunan25.55982 113.71602
264Hunan27.31478 111.715872
265Hunan27.22257 111.83645
266Hunan27.31478 111.71587
267Hunan27.31478 111.71587
268Hunan27.23382 111.83068
269Hunan27.29494 111.76303
270Hunan27.27205 111.69587
271Liaoning39.68004 122.89789
272Liaoning39.71416 122.97196
273Liaoning39.88909 124.09901
274Liaoning39.69624 122.93206
275Liaoning39.96288 124.22381
276Liaoning39.97018 124.24395
277Liaoning39.94946 124.18820
278Liaoning39.92736 124.12458
279Liaoning39.93187 124.15184
280Hainan18.41188 109.18473
281Hainan18.21360 109.10200
282Hainan18.36091 109.19870
283Hainan18.35456 109.20054
284Hainan18.21310 109.10550
285Hainan18.36428 109.19153
286Hainan18.41762 109.71702
287Hainan18.21360 109.10500
288Hainan18.24500 109.12230
289Hainan18.22130 109.10590
290Hainan18.35728 109.17184
291Jilin43.85878 125.54167
Table A4. Genetic identification of representative pathogenic isolates.
Table A4. Genetic identification of representative pathogenic isolates.
Strain No. GenBank
Accession No.
GeneGenus or SpeciesTop Match
(Accession)
Identity (%)Query Cover (%)E-Value
Alt1OK067695ITSAlternaria sp.MT951215.199.6990.0
AltSX241OK330459ITSAlternaria sp.OR964412.2100980.0
FSSC129OK330460ITSFusarium solaniOR123272.1100980.0
FSSX2571OK330461ITSF. solaniPV383491.1100980.0
FSCQ215OK330462ITSF. solaniPV383491.1100980.0
FVCQ231OK330463ITSF. virguliformeMZ854210.199.41000.0
FVSD211OK330464ITSF. virguliformeMZ854209.198.31000.0
FCSX567OK330465ITSF. chlamydosporumON242166.199.4990.0
RSCQ236OK330466ITSRhizoctonia solaniOL873284.199.71000.0
RSSC2244OK330467ITSR. solaniOL873284.199.41000.0
PenLN2522OK330468ITSPenicillium sp.KP900325.199.11000.0
PytSC255OK330469ITSPythium sp.KJ162354.199.8990.0
Plec1OK330470ITSPlectosphaerella sp.OR654246.199.4990.0
Chae1OK330471ITSChaetomium sp.MZ363862.199.4960.0
TriSX5531OK330472ITSTrichoderma sp.OK175851.199.71000.0
ASCO1OK330473ITSAscodesmis sp.MZ221602.199.81000.0
Humi1OK330474ITSHumicola sp.MW199085.199.4990.0
Botry1OK330475ITSBotryotrichum sp.MG770259.199.0980.0
Col1OK330476ITSColletotrichum sp.ON427165.199.81000.0
PhyCQ3681OK330477ITSPhytophthora sp.PP783476.199.51000.0
Asp1OK330478ITSAspergillus sp.NR_131292.199.81000.0
FOSX2933OR066400EF1αF. oxysporumMG735692.1100.01000.0
FOSX2731OR083083EF1αF. oxysporumHM057313.1100.01000.0
FOCQ3871OR083084EF1αF. oxysporumOQ130020.1100.01000.0
FOSC1110OR083085EF1αF. oxysporumMH698997.1100.01000.0
FOCQ3823OR083086EF1αF. oxysporumOQ130020.1100.01000.0
FOSC289OR083087EF1αF. oxysporumMW438342.1100.01000.0
FOSC1461OR083088EF1αF. oxysporumMW438342.1100.01000.0
FOSC1331OR083089EF1αF. oxysporumPQ654043.199.71000.0
FSSX2163OR083090EF1αF. solaniMN977911.1100.01000.0
FSSX2451OR083091EF1αF. solaniOQ511067.199.91000.0
FSCQ2422OR083092EF1αF. solaniOP184956.199.71000.0
FSCQ31042OR083093EF1αF. solaniON366424.1100.01000.0
Table A5. Virulence evaluation of representative isolates for each species obtained.
Table A5. Virulence evaluation of representative isolates for each species obtained.
SpeciesNo. of Plants for Each Disease Severity CategoryTotal of Plants InoculatedTotal of Isolates Tested
012345
R. solani2152221487418213
F. solani15738039211224017
F. oxysporum191108548231229720
Alternaria1149600302
Trichoderma2159220302
Pythium spp.01112700302
Phytophthora spp.03151100292
Penicillium spp.5146220292
Plectosphaerella spp.3181500272
Colletotrichum spp.6140600262

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Figure 1. Geographical distribution of 121 and 170 sampling sites where common bean samples with root rot symptoms were collected in 2016 and 2018, respectively. Five regions, marked with red ellipses, were used for comparing pathogen composition across different locations.
Figure 1. Geographical distribution of 121 and 170 sampling sites where common bean samples with root rot symptoms were collected in 2016 and 2018, respectively. Five regions, marked with red ellipses, were used for comparing pathogen composition across different locations.
Agriculture 15 01426 g001
Figure 2. The number of isolates per genus obtained from common bean root rot samples collected in China during 2016 and 2018. N represents the total number of identified isolates.
Figure 2. The number of isolates per genus obtained from common bean root rot samples collected in China during 2016 and 2018. N represents the total number of identified isolates.
Agriculture 15 01426 g002
Figure 3. Root rot symptoms on common bean (cv. English Red) following inoculation with different pathogenic isolates. (A,B) Rhizoctonia solani; (C,D) Fusarium oxysporum; (E,F) F. solani; (G) Penicillium spp.; (H) Pythium spp.; (I) Alternaria spp.; (J) Phytophthora spp.; (K) Plectosphaerella spp.; (L) Trichoderma spp. Test: plants inoculated with fungal or oomycete isolates; Mock: plants inoculated with sterile water.
Figure 3. Root rot symptoms on common bean (cv. English Red) following inoculation with different pathogenic isolates. (A,B) Rhizoctonia solani; (C,D) Fusarium oxysporum; (E,F) F. solani; (G) Penicillium spp.; (H) Pythium spp.; (I) Alternaria spp.; (J) Phytophthora spp.; (K) Plectosphaerella spp.; (L) Trichoderma spp. Test: plants inoculated with fungal or oomycete isolates; Mock: plants inoculated with sterile water.
Agriculture 15 01426 g003
Figure 4. Virulence analysis of pathogens causing root rot in common bean. For disease severity category data, non-parametric Kruskal–Wallis test were performed first, followed by post hoc pairwise comparisons using Dunn’s test. * represents p < 0.05, ** represents p < 0.01, **** represents p < 0.0001.
Figure 4. Virulence analysis of pathogens causing root rot in common bean. For disease severity category data, non-parametric Kruskal–Wallis test were performed first, followed by post hoc pairwise comparisons using Dunn’s test. * represents p < 0.05, ** represents p < 0.01, **** represents p < 0.0001.
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Figure 5. Frequency distribution of pathogen groups isolated from the common bean root rot samples across different geographical regions. First, transform the frequency distribution data into CLR (centered log-ratio) values. Then, perform multivariate PERMANOVA analysis to test for overall differences, followed by non-parametric Kruskal–Wallis tests with false discovery rate correction.
Figure 5. Frequency distribution of pathogen groups isolated from the common bean root rot samples across different geographical regions. First, transform the frequency distribution data into CLR (centered log-ratio) values. Then, perform multivariate PERMANOVA analysis to test for overall differences, followed by non-parametric Kruskal–Wallis tests with false discovery rate correction.
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Figure 6. Frequency distribution of pathogen groups isolated from the common bean root rot samples across different growth stages. First, transform the frequency distribution data into CLR (centered log-ratio) values. Then, perform multivariate PERMANOVA analysis to test for overall differences, followed by non-parametric Kruskal–Wallis tests with false discovery rate correction.
Figure 6. Frequency distribution of pathogen groups isolated from the common bean root rot samples across different growth stages. First, transform the frequency distribution data into CLR (centered log-ratio) values. Then, perform multivariate PERMANOVA analysis to test for overall differences, followed by non-parametric Kruskal–Wallis tests with false discovery rate correction.
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Table 1. Multivariate analysis of variance of frequency distribution of pathogen groups isolated from the common bean root rot samples across different geographical regions.
Table 1. Multivariate analysis of variance of frequency distribution of pathogen groups isolated from the common bean root rot samples across different geographical regions.
EffectDfSum of SqsR2FPr(>F)
Region213.690.434.470.002
Residual1218.370.57NANA
Total1432.061NANA
First, transform the frequency distribution data into CLR (centered log-ratio) values. Then, perform multivariate PERMANOVA analysis to test for overall differences.
Table 2. Geographic distribution and isolation frequency of Fusarium species in different regions of China.
Table 2. Geographic distribution and isolation frequency of Fusarium species in different regions of China.
Fusarium SpeciesNumber of Isolates
(Frequency %)
χ2p-Value
North ChinaEast China Southwest China
F. solani271 (47.3)45 (40.5)257 (42.8)0.550.8
F. oxysporum242 (42.2)63 (56.8) 340 (56.7) 2.70.3
F. virguliforme6(1.0)3 (2.7)3 (0.5)1.90.4
F. chlamydosporum54 (9.4)0 (0.0)0 (0.0)18.8<0.0001
χ2369.1105.3611.0
p-value<0.0001<0.0001<0.0001
Notes: For each column, the chi-square test was performed directly on the raw observed values, whereas row-wise comparisons were conducted using percentage-based data. The null hypothesis assumed (1) an even distribution of each Fusarium species across regions and (2) equal isolation frequencies among all species within each region.
Table 3. Multivariate analysis of variance of frequency distribution of pathogen groups isolated from the common bean root rot samples across different growth stages.
Table 3. Multivariate analysis of variance of frequency distribution of pathogen groups isolated from the common bean root rot samples across different growth stages.
EffectDfSum of SqsR2FPr(>F)
Growth stage213.690.434.470.006
Residual1218.370.57NANA
Total1432.061NANA
Notes: First, transform the frequency distribution data into CLR (centered log-ratio) values. Then, perform multivariate PERMANOVA analysis to test for overall differences.
Table 4. Abundance and isolation frequency of Fusarium species at different growth stages.
Table 4. Abundance and isolation frequency of Fusarium species at different growth stages.
Fusarium SpeciesNumber of Isolates (Frequency %)χ2p-Value
SeedlingPod-FilingMaturity
F. solani81 (34.2)154 (41.3)365 (48.9)2.60.3
F. oxysporum133 (56.1)193 (51.7)364 (48.7)0.50.8
F. virguliforme6 (2.5)5 (1.3)2 (0.3)1.80.4
F. chlamydosporum17 (7.2)21 (5.6)16 (2.1)2.70.3
χ2177.8285.8677.3
p-value<0.0001<0.0001<0.0001
Notes: For each column, the chi-square test was performed directly on the raw observed values, whereas row-wise comparisons were conducted using percentage-based data. The null hypothesis assumed (1) an even distribution of each Fusarium species across regions and (2) equal isolation frequencies among all species within each growth stage.
Table 5. Statistical analysis of pairwise co-infections between F. oxysporum, F. solani, and R. solani based on root infection frequencies.
Table 5. Statistical analysis of pairwise co-infections between F. oxysporum, F. solani, and R. solani based on root infection frequencies.
Primary PathogenPaired PathogenInfection StatusObserved (Expected)T-ValueStatistical Significance
F. solaniF. oxysporumDouble negative856−616.08p < 0.001
F. oxysporum
positive only
98−337.92
F. solani
positive only
185−424.92
Double positive473−233.08
F. solaniR. solaniDouble negative1036900.2p < 0.001
R. solani
positive only
5140.78
F. solani
positive only
358493.78
Double positive21377.22
R. solaniF. oxysporumDouble negative944824.99p < 0.001
F. oxysporum
positive only
10129.01
R. solani
positive only
450569.01
Double positive20888.09
Notes: The null hypothesis states that pathogen occurrences are mutually independent. Significance were calculated after Bonferroni correction (α = 0.001). T-value interpretation: negative values indicate observed frequencies significantly lower than expected under independence.
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Yang, L.; Lu, X.-H.; Wu, B.-M.; Zhong, Z.-M.; Li, S.-D. Spatiotemporal Profiling of the Pathogen Complex Causing Common Bean Root Rot in China. Agriculture 2025, 15, 1426. https://doi.org/10.3390/agriculture15131426

AMA Style

Yang L, Lu X-H, Wu B-M, Zhong Z-M, Li S-D. Spatiotemporal Profiling of the Pathogen Complex Causing Common Bean Root Rot in China. Agriculture. 2025; 15(13):1426. https://doi.org/10.3390/agriculture15131426

Chicago/Turabian Style

Yang, Li, Xiao-Hong Lu, Bo-Ming Wu, Zeng-Ming Zhong, and Shi-Dong Li. 2025. "Spatiotemporal Profiling of the Pathogen Complex Causing Common Bean Root Rot in China" Agriculture 15, no. 13: 1426. https://doi.org/10.3390/agriculture15131426

APA Style

Yang, L., Lu, X.-H., Wu, B.-M., Zhong, Z.-M., & Li, S.-D. (2025). Spatiotemporal Profiling of the Pathogen Complex Causing Common Bean Root Rot in China. Agriculture, 15(13), 1426. https://doi.org/10.3390/agriculture15131426

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