Pathogenic Interactions Between Pectobacterium spp. and Pseudomonas syringae in Potato as Influenced by Co-Inoculation and Temperature

Pathogenic Interactions Between Pectobacterium spp. and Pseudomonas syringae in Potato as Influenced by Co-Inoculation and Temperature | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Pathogenic Interactions Between Pectobacterium spp. and Pseudomonas syringae in Potato as Influenced by Co-Inoculation and Temperature Riad Saidu Koroma, Dugeri Rita Dooshima, Francess Sia Saquee, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8833264/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Potato production is challenged by bacterial soft rot and blackleg diseases caused by the Soft Rot Pectobacteriaceae (SRP), particularly Pectobacterium species. While Pseudomonas syringae , a common potato-related bacterium and well-studied plant pathogen, is known, its role in mixed infections with SRP under various environmental conditions remains unclear. This research examined the pathogenic potential of several Pectobacterium strains and P. syringae on potato tubers, explored how different strains interact during co-infection either synergistically or antagonistically, and analyzed how temperature influences disease development. Potato tubers were inoculated with either a single strain or a combination of P. syringae strains and then incubated at 5, 25, 30, and 35°C in controlled-environment chambers. Disease severity was measured by lesion area (mm²), and interaction outcomes were expressed as percentage changes relative to single-strain infections. Significant differences in pathogenicity were observed among SRP strains; Pectobacterium parmentieri and P. brasiliense exhibited high virulence, whereas other strains were weak or nonpathogenic when tested alone. Co-inoculation with P. syringae yielded strain-specific effects: it heightened disease severity in weakly pathogenic strains but decreased lesion formation in highly virulent ones, suggesting antagonism. Temperature markedly affected disease expression, with maximum virulence usually at 30°C. Co-inoculation also extended the temperature range conducive to disease development. These results illustrate that potato soft rot severity depends on bacterial strain type, microbial interactions, and temperature, underscoring the limitations of single-pathogen models and highlighting the importance of considering microbial communities and environmental factors in disease prediction and control. Soft rot Pectobacteriacea Polymicrobial infection synergistic and antagonistic interactions lesion area tissue maceration Solanum tuberosum Figures Figure 1 Figure 2 1. INTRODUCTION Potato ( Solanum tuberosum L.) is among the world’s most widely grown food crops, but bacterial diseases severely limit its production. These diseases lead to seed tuber rejection, reduced quality, and significant yield losses. Notably, the pectinolytic bacteria Dickeya and Pectobacterium are highly pathogenic, causing soft rot and blackleg worldwide. The global economic impact of these pathogens surpasses US $ 420 million annually, whereas in Europe, about €46 million is lost annually. In Israel, favorable conditions have led to a 20–25% reduction in yields (Dupuis et al. 2021 ; Toth et al. 2021 ; Degefu, 2024 ; Weronika Babinska-Wensierska et al. 2024 ). Given the limited efficacy of chemical control, alternative management strategies are urgently required (Mansfield et al. 2012 ; Szulta & Kornicka, 2023 ). Pectobacterium is characterized by its ability to break down plant cell walls through the secretion of plant cell wall-degrading enzymes (PCWDEs) (Hugouvieux-Cotte-Pattat et al. 2014). This genus is responsible for tuber loss in potatoes, both in the field and during storage (Portier et al.2020; Oulghazi et al. 2021), and it causes various symptoms, including soft rot, wilting, and blackleg, in potatoes and ornamental plants. Pseudomonas syringae is a key model organism for studying plant-microbe interactions and is responsible for causing serious plant diseases and economic damage globally (Mansfield et al. 2012 ; Chen et al. 2022 ). Different pathovars of P. syringae infect various plant species, leading to significant economic losses. It is among the most extensively studied plant pathogens and has contributed to a better understanding of host–microbe relationships, bacterial virulence factors, host adaptation, and microbial evolution, ecology, and epidemiology (Wang et al. 2023 ). Research on the molecular interactions between P. syringae and diverse host plants has significantly advanced our understanding of plant immune defenses and microbial pathogenicity. Although Pectobacterium spp. and P. syringae are both important and extensively studied plant pathogens, significant gaps remain in our understanding of their interactions. Specifically, there is a limited understanding of their co-infection dynamics, whether they interact synergistically or antagonistically in shared environments, and how their distinct pathogenic mechanisms have evolved (Davidsson et al. 2013 ; Katagiri et al. 2002 ). Additionally, there is limited knowledge of the in vitro pathogenicity of mixed infections involving different Pectobacterium spp. and P. syringae across various temperatures (Su et al., 2022 ). Understanding how plant-pathogenic bacteria interact is crucial for grasping disease progression and developing control methods. While many research efforts focus on single pathogens, mixed infections are common in agriculture and can yield outcomes that cannot be predicted by studying pathogens in isolation (Barrett et al., 2021 ). In potato diseases, interactions between Pectobacterium and Pseudomonas species are particularly intricate, involving both antagonistic and pathogenic relationships (Terletskiy & Lazarev, 2019 ; Aghdam et al. 2023 ). Some Pseudomonas strains can inhibit Pectobacterium through biocontrol, whereas others promote disease, with environmental factors such as temperature playing a significant role (Yang et al., 2024 ; Alattas et al., 2024 ; Höfte et al., 2021). Although evidence indicates both synergistic and antagonistic effects, the mechanisms underlying cooperation, competition, and increased pathogenicity among these bacteria remain poorly understood (Mahmoudi et al. 2011 ). Previous research has documented cases of synergistic disease enhancement. For example, studies have demonstrated that co-infection of plants with multiple viruses or with combinations of fungi and bacteria can increase disease severity (Bellah et al. 2023 ). However, the interactions between key potato pathogens, such as Pectobacterium species, and other common plant-associated bacteria remain poorly understood, particularly under varying environmental conditions. Temperature is a crucial abiotic factor that significantly affects both pathogen virulence and plant defense mechanisms. The optimal temperature for pathogen growth and infection varies widely across species, and temperature stress can weaken plant immunity, thereby increasing susceptibility to infection (Jee et al. 2020 ). To resolve the knowledge gap, the present study explores how selected strains of Pectobacterium and Pseudomonas interact pathogenically with potatoes across various temperatures. By clarifying these interactions, the study provides a better understanding of pathogenic interactions between Pectobacterium spp. and Pseudomonas syringae in potato under co-inoculation and varying temperatures, thereby supporting the development of more effective and sustainable control strategies. The study involved two main hypotheses, including (i) co-inoculation of Pectobacterium strains with P. syringae results in a synergistic increase in lesion development compared with each strain applied separately; and (ii) different temperature regimes influence disease progression in both individual and combined inoculations. The objectives of this study were: (i) to determine the pathogenicity of several Pectobacterium and P. syringae strains on potato tubers; (ii) to determine whether co-inoculation results in synergistic or antagonistic effects on lesion size; and (iii) to assess the influence of different temperature regimes on disease progression in both individual and combined inoculations. 2. MATERIALS AND METHODS 2.1 Experimental Facility, Material, Design, and Treatments The study was conducted in the Department of Agrobiotechnology's controlled laboratory facility at RUDN University’s Institute of Agriculture in Russia to evaluate bacterial pathogenicity and interactions with potato tubers under varying temperature conditions. The experimental materials were tubers of the Gala potato variety. The experiment was conducted using a completely randomized design with four replicates. All inoculations were performed in complete darkness to prevent light from affecting bacterial growth or host responses. Two complementary experiments were designed. In the first, tubers were inoculated with individual bacterial strains, co-inoculated with each Pectobacterium strain and Pseudomonas syringae Ps3004, or treated with a sterile buffer control, then incubated at a constant 30°C to evaluate potential synergistic or antagonistic effects. The second experiment examined how temperature influences pathogenicity using selected strains (F148, F126, KC, Ps3004) inoculated individually and in combination (F148 + Ps3004; F152 + Ps3004; F126 + Ps3004; and KC + Ps3004). The potato tubers were incubated at 5°C, 25°C, 30°C, and 35°C. A control group was kept at each temperature to ensure accurate comparison of disease responses. 2.2 Bacterial Inoculation and Colonization Experiment with Potato Tubers A total of four single-bacterial inoculation experiments were conducted using eight Pectobacterium strains and one Pseudomonas syringae Ps3004. The inoculation was performed according to the procedure described by Kõiv et al. ( 2015 ) and El-Hendawy et al. (2016). Accordingly, the strains were separately inoculated in liquid LB medium and cultured on suitable nutrient agar to produce enough biomass for the experiments. Cell suspensions were prepared in sterile buffer and adjusted to a uniform inoculum density by spectrophotometric measurement. The concentrations of the bacterial cultures were determined at an OD 600 of 0.5, and 100 µL of bacterial suspension was collected for each strain. Bacterial suspensions for each strain were prepared by mixing 0.4 mL of sterile water with 0.4 mL of bacterial culture. Potato tubers were surface-sterilized with NaClO (1%) for 5 min and thoroughly rinsed with sterile water three times and air-dried (Lee et al. 2019 ). The sterilized tubers were cut in half, and uniform wounds were created with a 15 mm cork borer (1 cm deep, 0.5 cm wide). The samples were sown in sterile plastic chambers lined with 2–3 layers of moist filter paper towels to maintain humidity and incubated at 5°C, 25°C, 30°C, and 35°C. A control group was kept at each temperature to ensure accurate comparison of disease responses. Four healthy potato tubers were then placed in sterile chambers with 2–3 layers of filter paper and were separately treated with 50 µL of bacterial culture suspension. The treated samples were kept at the various temperatures, and sterile water was added promptly to keep the filter paper moist until three days after inoculation. The control tubers were treated with a sterile medium solution. The experiment for each bacterial strain was conducted in four replicates. This method, adapted from Kõiv et al. ( 2015 ) and El-Hendawy et al. (2016), provided controlled, reproducible conditions for studying bacterial pathogenicity and interactions. For the co-bacterial inoculation experiment, equal volumes (0.4 mL) of each standardized suspension were mixed just before inoculation to ensure reliable bacterial interactions. The same standardized surface-sterilization protocol and inoculation technique described above were utilized. A total of 8 bacterial strains were mixed with Pseudomonas syringae Ps3004 to form the eight mixed bacterial complexes. Based on these 9 strains and the 8 mixed bacterial complexes, we compared the effects of single- and mixed-bacterial inoculation under diverse temperatures on the pathogenicity in potato. The experiment with 9 single-bacterial and 8 mixed-bacterial inoculations was performed using the same protocol as that described above. For temperature-dependent pathogenicity assays, selected treatments were additionally incubated at 5°C, 25°C, 30°C, and 35°C. Synergism experiments were mainly performed at 25°C or 30°C, which are optimal growth temperatures for these pathogens (Van der Wolf et al. 2017 ; Du Raan et al. 2016 ; Toth et al. 2021 ). Inoculation involved creating a standardized wound with a 15 mm Cork borer, then pipetting 0.5 mL of either a single bacterial strain or a mixed bacterial suspension into each wound. Control treatments received an equal volume of sterile buffer. Each treatment was replicated four times, and the entire experiment was repeated four times. Tubers were kept in darkness to prevent light effects. After three days, tubers were sliced to evaluate rotting and disease incidence. Table 1 The Pectobacterium strains and Pseudomonas syringae Ps3004 used in this work # Species Strain designation Origin (Year of Isolation) Taxonomic marker NCBI accession number 1 Pectobacterium brasiliense F126 Samara (2012) complete genome NZ_CP065031.1 2 Pectobacterium brasiliense F152 Moscow (2014) complete genome NZ_PJDM00000000.1 3 Pectobacterium versatile F131 (= F137) Kaluga (1998) complete genome NZ_PDVW01000000 4 Pectobacterium carotovorum F164(PB31) Moscow (2016) complete genome PJJA00000000.1 5 Pectobacterium parmentieri Pb20 Samara region (2012) complete genome NZ_PDDJ00000000 6 Pectobacterium parmentieri F148 (PB20) Moscow region (2013) 16S rRNA, BOX-PCR NZ_PDDJ00000000.1 7 Pectobacterium parmentieri F034 Kaluga Recombinase A (recA) gene MG518489.1 8 Pectobacterium sp. KC Moscow region (2025) 16S rRNA, BOX-PCR This study 9 Pseudomonas syringae Ps3004 Rostov region (2001) 16S rRNA This study 2.3 Lesion Measurement Disease severity was assessed using a standard lesion-based scoring system used in potato soft rot studies (Lee et al. 2019 ; Chung et al. 2017). After 72 h of incubation, tubers were sliced lengthwise through the inoculation point, and disease progression was evaluated by measuring lesion length and width (mm) with a digital caliper (Fig. 1 ). The lesion area was calculated as length × width (mm²) and served as the primary indicator of disease severity. This method provides a continuous, objective measure of tissue maceration and is widely used to evaluate the pathogenicity and virulence of Pectobacterium spp. and Pseudomonas syringae in potato tubers (Ge et al. 2021 ; Czajkowski et al. 2017 ; Kõiv et al. 2015 ). For each treatment, lesion areas on individual tubers (n = 4 per replicate) were measured, and the results were reported as mean values ± standard deviation. In co-inoculation experiments, microbial interactions were assessed by calculating the percentage change in lesion area compared to single-strain inoculations. Positive values signified synergistic effects, while negative values indicated antagonistic interactions. This standardized scoring system enabled reliable comparisons of disease severity across strains, treatments, and temperature conditions. Data Collection and Analysis After incubation, tubers were assessed for disease severity by measuring lesion size at the inoculation site. Each tuber was sliced through the center of the lesion (Fig. 1 ), and both lesion length and width were recorded in millimeters. The lesion area, obtained by multiplying length and width, served as a standardized metric of disease severity in square millimeters. For each treatment group, the average lesion area was calculated from the individual replicates. The data were compiled, and mean values and standard deviations were determined to assess variability. Treatment differences were analyzed using mean lesion area and percentage change. Results were summarized using descriptive statistics to highlight patterns and significant differences. Statistical evaluation involved ANOVA and post hoc tests to compare strains and treatments at an α level of 0.05 (Ge et al. 2021 ). Percentage Change Calculation: To determine the percentage change in lesion size (synergism and antagonism). We calculate the percentage increase or decrease in lesion area for co-inoculated treatments relative to the lesion area caused by the Pectobacterium strain alone using the formula: Where Lesion Area ms = mean lesion area of mixed inoculation (co-inoculated) strain treatment and Lesion Area ss = mean lesion area of single inoculation (single strain). Positive values indicate a synergistic effect (increased lesion size), whereas negative values indicate an antagonistic effect (reduced lesion size); see Table 2 . Negative values indicate decreased lesion sizes due to Ps3004; positive values indicate increased lesion sizes, reflecting strain-specific responses. 3. RESULT 3.1 Pathogenicity of Single and Co-inoculation Bacterial Strains The average lesion area (mm²) varied considerably among the SRP strains when inoculated alone and in combination with Pseudomonas syringae Ps3004. When applied individually, the virulence of each bacterial strain varied significantly. The results, summarized in Table 2 , indicate that not all strains were pathogenic under the experimental conditions. Pathogenic strains included P. parmentieri (Pb20, F034, F148), P. brasiliense (F152, F126), and P. syringae (Ps3004), all of which caused visible lesions on potato tubers. Lesion sizes ranged from an average of 103 mm 2 for Ps3004 to 316 mm 2 for P. brasiliense (F126). Non-pathogenic strains involved P. carotovorum (F164), which did not produce any visible lesions; P. versatile strain F131, and Pectobacterium sp. (KC), which failed to produce lesions at + 50°C and + 25°C; and Pectobacterium sp. (KC), which did not produce lesions at + 50°C. The control (K) showed no reaction, confirming that the lesions were caused by bacterial activity. Co-inoculating weakly pathogenic Pectobacterium strains with P. syringae Ps3004 significantly increased lesion severity, especially with P. carotovorum (F164), P. parmentieri (F148), P. versatile strain F131, and Pectobacterium sp. (KC). A moderate synergistic effect was observed with the P. versatile strain F131, which increased lesion area by 94.4% when combined with Ps3004. The combination of F148 and Ps3004 caused a 9.6% increase, whereas Pectobacterium sp (KC) increased by 7.4%, and P. carotovorum F164 exhibited a substantial increase of 4300%. Conversely, co-inoculating highly virulent Pectobacterium strains with P. syringae Ps3004 reduced lesion severity, demonstrating an antagonistic effect. This was particularly evident in P. parmentieri (Pb20, F034) and P. brasiliense (F126). For example, combining Pb20 with Ps3004 resulted in an average lesion area of 432 mm 2 , a 32.2% reduction compared with Pb20 alone. The pairing of F034 and Ps3004 resulted in a lesion area of 448 mm 2 , a 43.8% decrease over F034 alone. Figure 2 displays soft rot symptoms in potato tubers inoculated with different bacterial strains: Pseudomonas syringae Ps3004 (a), Pectobacterium sp. KC (b), Pectobacterium brasiliense F148 (c), and Pectobacterium brasiliense F126 (d). The tubers were kept in darkness at 30°C for 48 h. The images highlight the strain-dependent variations in tissue maceration, discoloration, and exudate formation. Table 2 Effects of co-inoculation on lesion size caused by pectotytic bacteria. (A) Shows the average lesion area (mm 2 ) after co-inoculation with Ps3004. (B) Displays the average squared lesion area (mm 2 ) for single-strain inoculation. The percentage change indicates the change in lesion area when Ps3004 is combined with the strain relative to the strain alone. Strains Species Average squared lesion area (mm 2 ) + Ps3004 (A) Av. squared lesion area (mm 2 ) (B) Percentage change (%) Pb20 P. parmentieri 293 432 -139(-32.2%) F034 P. parmentieri 252 448 -196(-43.8%) F148 P. parmentieri 286 261 + 25(9.6%) F152 P. brasiliense 308 353 -45(-12.7%) F164 P. carotovorum 66 1.5 + 64.5(4300%) F126 P. brasiliense 316 357 -41(-11.5%) F131 P. versatile 241 124 + 117(94.4%) KC Pectobacterium sp 246 229 + 17(7.4%) 3.2 Influence of Temperature on Pathogenicity Temperature played a crucial role in lesion development and the interactions among the tested strains, emphasizing its importance in controlling soft rot severity. As indicated in Table 3 , no lesions formed at 5°C, showing that bacterial activity was suppressed at low temperatures. Conversely, disease severity increased significantly at 25°C and peaked at 30°C for the most aggressive strains, particularly Pectobacterium brasiliense F126 and P. parmentieri F148. The decrease in lesion size at 35°C points to thermal stress or a decline in virulence-related processes at temperatures above the optimum. Co-inoculation changed how temperature affects pathogen responses. Combining P. brasiliense F126 with Pseudomonas syringae Ps3004 extended the temperature range over which pathogenic effects are observed, maintaining visible lesions from 25°C to 35°C (Table 3 ). Similarly, Pectobacterium sp. KC showed increased lesion formation at intermediate temperatures when co-inoculated, despite having limited virulence on its own. These results imply that interspecies interactions can help buffer against environmental stress and alter virulence levels. The temperature-dependent patterns align with earlier research showing that enzyme activity, motility, and quorum sensing in Soft Rot Pectobacteriaceae are most active at moderate temperatures and decrease under extreme heat or cold (Czajkowski et al., 2017 ; Jee et al., 2020 ; Toth et al., 2021 ). Table 3 Temperature-dependent pathogenicity of selected bacterial strains and co-inoculations on potato tubers, highlighting the synergistic and antagonistic effects of co-inoculation with Pseudomonas syringae Ps3004, P. brasiliense (F126, F152), P. versatile strain F131, P . parmentieri (Pb20, F034, F148), P. carotovorum (F164), and Pectobacterium sp. (KC), with Pseudomonas syringae Ps3004 Pseudomonas syringae (Ps3004) P. Brasiliense (F152) P. parmentier (F148) Pectobacterium sp. KC Mean scores (mm²) 5°C 25°C 30°C 35°C - - - + 0 272 270 375 + - - + 0 198 308 480 - + - - 0 0 972 779 + + - - 0 210 462 306 - - + - 0 180 273 460 + - + - 0 295 268 338 + - - - 0 0 476 224 Symbols (+) and (–) indicate the presence and absence of a bacterial strain during inoculation, respectively Lesion development (mm²) of potato tubers inoculated with selected bacterial strains and their combinations under different temperature regimes (5, 25, 30, and 35°C) (Table 3 ). Values are presented as mean ± SD (n = 4). For each temperature, treatment means were compared using one-way ANOVA; means that differed by lowercase letter within the same temperature column were considered significantly different based on post hoc multiple comparisons (P < 0.05). 4. DISCUSSION This study presents experimental evidence that disease severity in potato tubers is influenced by bacterial strain identity, interspecific interactions, and temperature. Notable differences in virulence were found among Soft Rot Pectobacterium bacteria and Pseudomonas syringae , underscoring the complexity of mixed bacterial infections. Pectobacterium parmentieri (Pb20, F034, F148), P. brasiliense (F126, F152), and P. syringae (Ps3004) were pathogenic when tested individually, while P. carotovorum (F164) was non-pathogenic under the experimental conditions, suggesting strain-specific or environment-dependent virulence. The lack of lesions in control treatments confirms that lesion formation was caused solely by bacterial activity. 4.1 Pathogenic Variability among SRP Strains The present study revealed pronounced pathogenic variability among Soft Rot Pectobacteriaceae (SRP) strains, confirming that virulence within this group is highly strain dependent and cannot be inferred solely from species identity. When inoculated individually, Pectobacterium parmentieri (Pb20, F034, F148) and P. brasiliense (F126, F152) consistently induced large lesions on potato tubers, whereas P. carotovorum (F164), P. versatile (F131), and Pectobacterium sp. (KC) exhibited weak or negligible pathogenicity under the same experimental conditions. These findings clearly demonstrate substantial intra- and interspecific heterogeneity in pathogenic potential among SRP strains (Czajkowski et al. 2011; Toth et al. 2021 ). Such variability is well documented in SRP pathosystems and reflects differences in pectinolytic enzyme production, quorum-sensing regulation, stress tolerance, and environmental responsiveness rather than taxonomic classification alone (Czajkowski et al. 2011; Toth et al. 2021 ). Previous studies have shown that isolates of P. brasiliense and P. parmentieri differ markedly in aggressiveness, maceration kinetics, and temperature optima, even when recovered from the same host or geographic region (Golanowska et al. 2017 ; van der Wolf et al. 2017 ; Zhou et al. 2024 ). Our results are consistent with these reports, as strains within the same species exhibited contrasting pathogenic behaviors. The absence of lesion formation by P. carotovorum F164 and Pectobacterium sp. KC during individual inoculation indicates that some SRP strains can survive in potato tissues as low-virulence or opportunistic colonizers. Their ability to cause disease appears to depend on specific environmental conditions or the presence of other microbes. Similar findings have been reported for SRP isolates with reduced pectate lyase activity or compromised virulence gene expression, which induce disease only when additional stressors or interacting microorganisms are present (Davidsson et al. 2013 ; Czajkowski et al. 2017 ). Overall, the variability in pathogenic traits underscores the limitations of single-strain or species-level models for predicting the risk of potato soft rot. Disease outcomes are influenced by a mix of strain-specific characteristics, environmental factors, and microbial interactions. Understanding this diversity is essential for improving disease diagnostics, refining risk assessment models, and developing targeted management strategies that account for the ecological complexity of SRP populations in potato production systems. 4.2 Synergistic and Antagonistic Interactions with Pseudomonas syringae Co-inoculation experiments revealed that interactions between SRP strains and P. syringae Ps3004 vary and are not always mutually beneficial. Some combinations led to more severe lesions, while others reduced disease severity, indicating antagonistic effects (Fig. 3). The most pronounced increases were observed with strains that were weakly pathogenic or non-pathogenic alone, such as P. brasiliense F137, P . sp. KC, and P. carotovorum F164. These results align with previous studies indicating that mixed bacterial infections can amplify disease severity through mechanisms such as tissue maceration, changes in quorum sensing, or suppression of host defenses (Degefu et al. 2021). In contrast, the antagonistic effects observed with P. parmentieri (Pb20, F034) and P. brasiliense (F126) suggest competitive exclusion or interference among coexisting bacteria. Similar results have been reported in SRP pathosystems, where competition for nutrients, space, or oxygen can reduce virulence in mixed infections (Arif et al. 2022). These responses indicate that mixed infections do not necessarily lead to more severe disease, as outcomes depend on strain-specific traits and ecological factors. 4.3 Temperature as a Modulator of Pathogenicity and Interaction Outcomes Temperature significantly influenced disease expression, reaffirming its role as a key environmental factor affecting SRP virulence. The highest lesion development for P. syringae Ps3004 and P. brasiliense F126 occurred at 30°C, with a decline at 35°C. The combination of P. brasiliense F126 and P. syringae Ps3004 caused disease over a wider temperature range (25°C to 35°C), with lesion severity increasing with temperature. Similarly, P. parmentieri F148 caused disease across this range when inoculated alone, exhibiting high pathogenicity at 30°C and reduced pathogenicity at 35°C. The strain KC ( Pectobacterium sp) showed linear disease symptoms only between 25°C and 35°C when co-inoculated with P. syringae (Ps3004). These temperature-dependent behaviors illustrate the complex interaction between environmental conditions and pathogen activity in disease development. This aligns with earlier research indicating that SRP virulence factors, including pectolytic enzyme activity and motility, are most effective at moderate temperatures and decrease under thermal stress (Czajkowski et al. 2017 ; Jee et al. 2020 ; Toth et al. 2021 ). For P. brasiliense (F126), pectolytic activity of Pectobacterium diminished at 35°C when co-inoculated with P. syringae (Ps3004), in contrast to P. parmentieri (F148). Notably, co-inoculating P. brasiliense F126 with Ps3004 broadened the temperature range for disease development, demonstrating that interspecific interactions can mitigate environmental limits. Similar temperature-dependent effects on disease severity have been observed in mixed infections involving Dickeya and Pectobacterium species, where cooperative interactions intensify under favorable climatic conditions (van der Wolf et al. 2021 ; Degefu, 2024 ; Jee et al. 2020 ; Lebecka, et al. 2018 ). These results are especially significant given predictions of climate change, which suggest an increase in conditions conducive to SRP outbreaks. 5. CONCLUSION This study demonstrated that bacterial strain, interspecific interactions from co-inoculation involving Pectobacterium spp. and Pseudomonas syringae , and temperature influence disease development (pathogenicity and disease severity) in potato tubers, findings that could be exploited to develop more effective and sustainable control strategies and to improve potato breeding for disease resistance. The combined evidence from pathogenicity, interaction, and temperature experiments indicates that relying solely on single-pathogen models for disease risk assessments might underestimate the complexity of SRP epidemics. Mixed infections can either worsen or suppress disease outcomes depending on strain combinations and environmental factors. Enhancing detection of latent infections and understanding interaction-driven disease dynamics will be crucial for sustainable management. Pathogenicity varied significantly among SRP strains, reflecting substantial intra-species heterogeneity. Co-inoculation with Pseudomonas syringae Ps3004 yielded both synergistic and antagonistic effects, demonstrating that mixed infections do not always lead to increased disease severity. Temperature heavily influenced pathogenic outcomes, with peak virulence typically at 30°C and diminished disease expression at higher temperatures. Findings suggest that SRP epidemics are driven by strain-specific traits, microbial interactions, and environmental factors rather than a single cause. These findings support the importance of integrated disease management strategies that account for ecological complexity, especially amid changing climate conditions. Future research should explore the mechanisms underlying interaction-driven virulence, improve early detection of latent infections, and develop predictive models that incorporate microbial interactions and environmental influences. Declarations Acknowledgements The authors acknowledge the support given bytheDepartment of Agrobiotechnology, Agrarian-Technological Institute, RUDN University, 117198 Moscow, Russia. Ethics approval and consent to participate Not applicable. Consent for publication All authors agree with the submission of this manuscript to Plant Methods. Funding The research was supported by the Russian Ministry of Education and Science Project FSSF-2024-0063. Data Availability Statement The data that support the findings of this study are available from the corresponding author upon reasonable request. Competing interests: The authors have declaredno known conflicts of interest at the time of writing this article. The authors affirm that they have no recognized competing financial interests or personal connections that could have appeared to affect the work presented in this paper. Competing interests The authors declare that they have no competing interests. References Aghdam, N. M. N., Baghaee-Ravari, S., & Shiri, A. (2023). Antimicrobial capacity of Pseudomonas brassicacearum strain EnPb against potato soft rot agent. European Journal of Plant Pathology, 165(2), 215–231. https://doi.org/10.1007/s10658-022-02600-z Alattas, H., Glick, B. R., Murphy, D. V., & Scott, C. (2024). Harnessing Pseudomonas spp. for sustainable plant crop protection. Frontiers in microbiology, 15, 1485197. https://doi.org/10.3389/fmicb.2024.1485197 . Arif, M., Busot, G. Y., Mann, R., Rodoni, B., & Stack, J. P. (2021). Field-deployable recombinase polymerase amplification assay for specific, sensitive and rapid detection of the US select agent and toxigenic bacterium, Rathayibacter toxicus. Biology, 10(7), 620. https://doi.org/10.3390/biology10070620 Babinska-Wensierska, W., Motyka-Pomagruk, A., Fondi, M., Misztak, A. E., Mengoni, A., & Lojkowska, E. (2024). Differences in the constituents of bacterial microbiota of soils collected from two fields of diverse potato blackleg and soft rot diseases incidences, a case study. Scientific Reports, 14(1), 18802. https://doi.org/10.1038/s41598-024-69213-w Barrett, L. G., Zala, M., Mikaberidze, A., Alassimone, J., Ahmad, M., McDonald, B. A., & Sánchez-Vallet, A. (2021). Mixed infections alter transmission potential in a fungal plant pathogen. Environmental microbiology, 23(4), 2315–2330. https://doi.org/10.1111/1462-2920.15417 Bellah, H., Seiler, N. F., & Croll, D. (2023). Divergent outcomes of direct conspecific pathogen strain interaction and plant co-infection suggest consequences for disease dynamics. Microbiology Spectrum, 11(2), e04443-22. https://doi.org/10.1128/spectrum.04443-22 Chen, H., Chen, J., Zhao, Y., Liu, F., & Fu, Z. Q. (2022). Pseudomonas syringae pathovars. Trends in Microbiology, 30(9). https://doi.org/10.1016/j.tim.2022.03.002 Chung YongSuk, C. Y., Kim ChangSoo, K. C., & Jansky, S. (2017). New source of bacterial soft rot resistance in wild potato (Solanum chacoense) tubers. https://doi.org/10.1007/s10722-017-0487-3 Czajkowski, R. (2011). Ecology and control of Dickeya spp. in potato. Netherlands Institute of Ecology. https://hdl.handle.net/1887/17810 Czajkowski, R., Kaczyńska, N., Jafra, S., Narajczyk, M., & Lojkowska, E. (2017). Temperature-responsive genetic loci in pectinolytic plant pathogenic Dickeya solani. Plant Pathology, 66(4), 584–594. https://doi.org/10.1111/ppa.12618 Davidsson, P. R., Kariola, T., Niemi, O., & Palva, E. T. (2013). Pathogenicity of and plant immunity to soft rot pectobacteria. Frontiers in plant science, 4, 191. https://doi.org/10.3389/fpls.2013.00191 Degefu, Y., 2021. Co-occurrence of latent Dickeya and Pectobacterium species in potato seed tuber samples from northern Finland: Co-colonization of latent Dickeya and Pectobacterium species in potato seed lots. Agricultural and Food Science, 30(1), pp.1–7. https://doi.org/10.23986/afsci.101446 Degefu, Y., 2024. Lesson from the emergence, spread and decline of Dickeya solani, the virulent potato blackleg and soft rot bacterial pathogen in Finland. Journal of phytopathology, 172(2), p.e13282. https://doi.org/10.1111/jph.13282 Du Raan, S., Coutinho, T. A., & Van der Waals, J. E. (2016). Cardinal temperature differences, determined in vitro, between closely related species and subspecies of pectinolytic bacteria responsible for blackleg and soft rot on potatoes. European Journal of Plant Pathology, 144(2), 361–369. https://doi.org/10.1007/s10658-015-0773-x Dupuis, B., Nkuriyingoma, P., & Van Gijsegem, F. (2021). Economic impact of Pectobacterium and Dickeya species on potato crops: A review and case study. In Plant diseases caused by dickeya and pectobacterium species (pp. 263–282). Cham: Springer International Publishing. https://doi.org/10.1007/978-3-030-61459-1_8 El-Hendawy, H. H., & Abo-Elyousr, K. A. M. (2016). Combination of different antagonistic bacteria to control of potato blackleg disease caused by Pectobacterium atrosepticum under greenhouse and field conditions. Ge, T., Ekbataniamiri, F., Johnson, S. B., Larkin, R. P., & Hao, J. (2021). Interaction between Dickeya dianthicola and Pectobacterium parmentieri in potato infection under field conditions. Microorganisms, 9(2), 316. https://doi.org/10.3390/microorganisms9020316 Golanowska, M., Kielar, J., & Lojkowska, E. (2017). The effect of temperature on the phenotypic features and the maceration ability of Dickeya solani strains isolated in Finland, Israel and Poland. European Journal of Plant Pathology, 147, 803–817. https://doi.org/10.1007/s10658-016-1044-1 Höfte, M. (2021). The use of Pseudomonas spp. as bacterial biocontrol agents to control plant disease. In Microbial bioprotectants for plant disease management. Burleigh Dodds. http://dx.doi.org/10.19103/AS.2021.0093.11 Hugouvieux-Cotte‐Pattat, N., Condemine, G., & Shevchik, V. E. (2014). Bacterial pectate lyases, structural and functional diversity. Environmental microbiology reports, 6(5), 427–440. https://doi.org/10.1111/1758-2229.12166 Jee, S., Choi, J. G., Lee, Y. G., Kwon, M., Hwang, I., & Heu, S. (2020). Distribution of Pectobacterium species isolated in South Korea and comparison of temperature effects on pathogenicity. The plant pathology journal, 36(4), 346. https://orcid.org/0000-0001-7834-4303 Katagiri, F., Thilmony, R., & He, S. Y. (2002). The Arabidopsis thaliana-Pseudomonas syringae interaction. The Arabidopsis Book/American Society of Plant Biologists, 1, e0039. https://doi.org/10.1199/tab.0039 Kõiv, V., Roosaare, M., Vedler, E., Ann Kivistik, P., Toppi, K., Schryer, D. W., … Mäe, A. (2015). Microbial population dynamics in response to Pectobacterium atrosepticum infection in potato tubers. Scientific reports, 5(1), 11606. https://doi.org/10.1038/srep11606 Kõiv, V., Roosaare, M., Vedler, E., Ann Kivistik, P., Toppi, K., Schryer, D. W., … Mäe, A. (2015). Microbial population dynamics in response to Pectobacterium atrosepticum infection in potato tubers. Scientific reports, 5(1), 11606. https://doi.org/10.1038/srep11606 Lebecka, R., Flis, B., & Murawska, Z. (2018). Comparison of temperature effects on the in vitro growth and disease development in potato tubers inoculated with bacteria Pectobacterium atrosepticum, P carotovorum subsp. carotovorum and Dickeya solani. Journal of Phytopathology, 166(9), 654–662. https://doi.org/10.1111/jph.12728 Lee, U., Silva, R. R., Kim, C., Kim, H., Heo, S., Park, I. S., … Chung, Y. S. (2019). Image analysis for measuring disease symptom to bacterial soft rot in potato. American Journal of Potato Research, 96(3), 303–313. https://doi.org/10.1007/s12230-019-09717-8 Mahmoudi, E., Ahmadi, A., Sayed-Tabatabaei, B. E., Ghobadi, C., Akhavan, A., Hasanzadeh, N., & Venturi, V. (2011). A novel AHL-degrading rhizobacterium quenches the virulence of Pectobacterium atrosepticum on potato plant. Journal of Plant Pathology, 587–594. http://dx.doi.org/10.4454/jpp.v93i3.1226 Mansfield, J., Genin, S., Magori, S., Citovsky, V., Sriariyanum, M., Ronald, P., … Foster, G. D. (2012). Top 10 plant pathogenic bacteria in molecular plant pathology. Molecular plant pathology, 13(6), 614–629. https://doi.org/10.1111/j.1364-3703.2012.00804.x Oulghazi, S., Moumni, M., Khayi, S., Robic, K., Sarfraz, S., Lopez-Roques, C., Vandecasteele, C., & Faure, D. (2020). Diversity of Pectobacteriaceae Species in Potato Growing Regions in Northern Morocco. Microorganisms, 8(6), 895. https://doi.org/10.3390/microorganisms8060895 Portier, P., Pédron, J., Taghouti, G., Dutrieux, C., & Barny, M. A. (2020). Updated taxonomy of Pectobacterium genus in the CIRM-CFBP bacterial collection: when newly described species reveal “old” endemic population. Microorganisms, 8(9), 1441. https://doi.org/10.3390/microorganisms8091441 Su, Z., Liu, X., Guo, Q., Xuan, L., Lu, X., Dong, L., … & Ma, P. (2022). Insights into complex infection by two Pectobacterium species causing potato blackleg and soft rot. Microbiological Research, 261, 127072. https://doi.org/10.1016/j.micres.2022.127072 Szulta, S., & Kornicka, A. (2023). Pectobacterium and Dickeya genus—A review on structural variations of O-polysaccharides and their role in the pathogenic process of plants. Plant Pathology, 72(6), 998–1010. https://doi.org/10.1111/ppa.13728 Terletskiy, V. P., & Lazarev, A. M. (2019). On genotyping bacterial strains of the genera Pectobacterium and Pseudomonas: pathogens of bacterioses in potatoes. Cytology and Genetics, 53(3), 212–218. https://doi.org/10.3103/S0095452719030058 Toth, I. K., Barny, M. A., Brurberg, M. B., Condemine, G., Czajkowski, R., Elphinstone, J. G., … Yedidia, I. (2021). Pectobacterium and Dickeya: environment to disease development. In Plant diseases caused by Dickeya and Pectobacterium species (pp. 39–84). Cham: Springer International Publishing. https://doi.org/10.1007/978-3-030-61459-1_3 van der Wolf, J. M., Acuña, I., De Boer, S. H., Brurberg, M. B., Cahill, G., Charkowski, A. O., … Yedidia, I. (2021). Diseases caused by Pectobacterium and Dickeya species around the world. In Plant diseases caused by Dickeya and Pectobacterium species (pp. 215–261). Cham: Springer International Publishing. https://doi.org/10.1007/978-3-030-61459-1_7 Van der Wolf, J. M., De Haan, E. G., Kastelein, P., Krijger, M., De Haas, B. H., Velvis, H., … Van Der Zouwen, P. S. (2017). Virulence of Pectobacterium carotovorum subsp. brasiliense on potato compared with that of other Pectobacterium and Dickeya species under climatic conditions prevailing in the Netherlands. Plant Pathology, 66(4), 571–583. https://doi.org/10.1111/ppa.12600 Wang, T., Hua, C., & Deng, X. (2023). c-di-GMP signaling in Pseudomonas syringae complex. Microbiological Research, 275, 127445. https://doi.org/10.1016/j.micres.2023.127445 Yang, Z., Liu, T., Fan, J., Chen, Y., Wu, S., Li, J., … Tang, Q. (2024). Biocontrol agents modulate phyllosphere microbiota interactions against pathogen Pseudomonas syringae. Environmental Science and Ecotechnology, 21, 100431. https://doi.org/10.1016/j.ese.2024.100431 Zhou, J., Hu, M., & Zhang, L. (2024). Dickeya Diversity and Pathogenic Mechanisms. Annual Review of Microbiology, 78(1), 621–642. https://doi.org/10.1146/annurev-micro-041222-012242 Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8833264","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":593441788,"identity":"2d816146-8292-408f-bee0-a30ce21b1b47","order_by":0,"name":"Riad Saidu Koroma","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA2klEQVRIiWNgGAWjYBACNmaGxAcfeP7J8bM3HwByidDCx97w2HCGzAFjyZ5jCcRpkeM5+Eyax+ZA4oYZPgbEaWGTSE6Q4Mm5w7hBgufbZ56yOwzmMxIIaUlLMJA484zZXLp382yec88YZG4Q1JKTkGDYw8xmOefsZmbetsMMEhIEteR/OJD4j5nH4EbOYyK18BxIbDjAc1gCqIWZSC3sDcmMDTxpBsBANmacc+4wjwTPA/xa5JsZ0n//4bGp72dvfszwpuywnAQ7AVswAA+J6kfBKBgFo2AUYAMAqx9DL2OCTo4AAAAASUVORK5CYII=","orcid":"","institution":"Peoples' Friendship University of Russia","correspondingAuthor":true,"prefix":"","firstName":"Riad","middleName":"Saidu","lastName":"Koroma","suffix":""},{"id":593441789,"identity":"eb1c7697-4a2e-4325-8e0f-054cf7cbeb35","order_by":1,"name":"Dugeri Rita Dooshima","email":"","orcid":"","institution":"Peoples' Friendship University of Russia","correspondingAuthor":false,"prefix":"","firstName":"Dugeri","middleName":"Rita","lastName":"Dooshima","suffix":""},{"id":593441790,"identity":"91dd62e3-170d-4613-930e-bd80b0774335","order_by":2,"name":"Francess Sia Saquee","email":"","orcid":"","institution":"Eastern Technical University","correspondingAuthor":false,"prefix":"","firstName":"Francess","middleName":"Sia","lastName":"Saquee","suffix":""},{"id":593441791,"identity":"434540e7-4083-4930-8b90-18d0ac2f0de0","order_by":3,"name":"Elena Pakina","email":"","orcid":"","institution":"Peoples' Friendship University of Russia","correspondingAuthor":false,"prefix":"","firstName":"Elena","middleName":"","lastName":"Pakina","suffix":""},{"id":593441792,"identity":"612976fa-0a68-4d32-aaea-f0a5aa502b6f","order_by":4,"name":"Aleksandr Nikolaevich Ignatov","email":"","orcid":"","institution":"Peoples' Friendship University of Russia","correspondingAuthor":false,"prefix":"","firstName":"Aleksandr","middleName":"Nikolaevich","lastName":"Ignatov","suffix":""},{"id":593441793,"identity":"fd76c437-88a4-4bb5-9abf-d7b1155f18e1","order_by":5,"name":"Prince Emmanuel Norman","email":"","orcid":"","institution":"Sierra Leone Agricultural Research Institute (SLARI)","correspondingAuthor":false,"prefix":"","firstName":"Prince","middleName":"Emmanuel","lastName":"Norman","suffix":""}],"badges":[],"createdAt":"2026-02-09 17:53:14","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8833264/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8833264/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":103059674,"identity":"4bf55bf5-7bb0-4c7c-af0b-ce4234ded9b2","added_by":"auto","created_at":"2026-02-20 09:41:36","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":590335,"visible":true,"origin":"","legend":"\u003cp\u003eA longitudinal cross-section of a Gala potato tuber, illustrating lesion size after three days of inoculation with \u003cem\u003eP. parmentieri\u003c/em\u003e (Pb20) and \u003cem\u003eP. parmentieri\u003c/em\u003e (F034)\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-8833264/v1/c527a8cbb44cc3aaf2676684.png"},{"id":103059728,"identity":"9ebe2a83-c25a-4509-88d9-63dff40b1330","added_by":"auto","created_at":"2026-02-20 09:41:41","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":827700,"visible":true,"origin":"","legend":"\u003cp\u003eSoft rot development in potato tubers following inoculation with different bacterial strains: (a) \u003cem\u003ePseudomonas syringae\u003c/em\u003e Ps3004 @ 35\u003csup\u003e0\u003c/sup\u003eC, (b)\u003cem\u003e Pectobacterium\u003c/em\u003e sp. KC @ 35\u003csup\u003e0\u003c/sup\u003eC, (c)\u003cem\u003e P. parmentieri\u003c/em\u003e F148 @ 35\u003csup\u003e0\u003c/sup\u003eC,\u003cem\u003e \u003c/em\u003e(d) P. brasiliense F126 @ 30\u003csup\u003e0\u003c/sup\u003eC, (e) F148 + Ps3004 @ 35\u003csup\u003e0\u003c/sup\u003eC, (f) F152 @ 30\u003csup\u003e0\u003c/sup\u003eC, (g) F148 + Ps3004 @ 30\u003csup\u003e0\u003c/sup\u003eC, (h) Pectobacterium sp. KC @ 30\u003csup\u003e0\u003c/sup\u003eC\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-8833264/v1/129fdee2a7d9e0ddcd0b55a4.png"},{"id":103819787,"identity":"4e8cf27b-6028-4cbd-a2cf-0d5fba29ca3a","added_by":"auto","created_at":"2026-03-03 09:57:39","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3116233,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8833264/v1/5d1d4bc7-7df6-441a-8a7e-17551f5bd362.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Pathogenic Interactions Between Pectobacterium spp. and Pseudomonas syringae in Potato as Influenced by Co-Inoculation and Temperature","fulltext":[{"header":"1. INTRODUCTION","content":"\u003cp\u003ePotato (\u003cem\u003eSolanum tuberosum\u003c/em\u003e L.) is among the world\u0026rsquo;s most widely grown food crops, but bacterial diseases severely limit its production. These diseases lead to seed tuber rejection, reduced quality, and significant yield losses. Notably, the pectinolytic bacteria Dickeya and Pectobacterium are highly pathogenic, causing soft rot and blackleg worldwide. The global economic impact of these pathogens surpasses US\u003cspan\u003e$\u003c/span\u003e420\u0026nbsp;million annually, whereas in Europe, about \u0026euro;46\u0026nbsp;million is lost annually. In Israel, favorable conditions have led to a 20\u0026ndash;25% reduction in yields (Dupuis et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Toth et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Degefu, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Weronika Babinska-Wensierska et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Given the limited efficacy of chemical control, alternative management strategies are urgently required (Mansfield et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Szulta \u0026amp; Kornicka, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). \u003cem\u003ePectobacterium\u003c/em\u003e is characterized by its ability to break down plant cell walls through the secretion of plant cell wall-degrading enzymes (PCWDEs) (Hugouvieux-Cotte-Pattat et al. 2014). This genus is responsible for tuber loss in potatoes, both in the field and during storage (Portier et al.2020; Oulghazi et al. 2021), and it causes various symptoms, including soft rot, wilting, and blackleg, in potatoes and ornamental plants.\u003c/p\u003e \u003cp\u003e \u003cem\u003ePseudomonas syringae\u003c/em\u003e is a key model organism for studying plant-microbe interactions and is responsible for causing serious plant diseases and economic damage globally (Mansfield et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Chen et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Different pathovars of \u003cem\u003eP. syringae\u003c/em\u003e infect various plant species, leading to significant economic losses. It is among the most extensively studied plant pathogens and has contributed to a better understanding of host\u0026ndash;microbe relationships, bacterial virulence factors, host adaptation, and microbial evolution, ecology, and epidemiology (Wang et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Research on the molecular interactions between \u003cem\u003eP. syringae\u003c/em\u003e and diverse host plants has significantly advanced our understanding of plant immune defenses and microbial pathogenicity.\u003c/p\u003e \u003cp\u003eAlthough \u003cem\u003ePectobacterium\u003c/em\u003e spp. and \u003cem\u003eP. syringae\u003c/em\u003e are both important and extensively studied plant pathogens, significant gaps remain in our understanding of their interactions. Specifically, there is a limited understanding of their co-infection dynamics, whether they interact synergistically or antagonistically in shared environments, and how their distinct pathogenic mechanisms have evolved (Davidsson et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Katagiri et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). Additionally, there is limited knowledge of the in vitro pathogenicity of mixed infections involving different Pectobacterium spp. and P. syringae across various temperatures (Su et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eUnderstanding how plant-pathogenic bacteria interact is crucial for grasping disease progression and developing control methods. While many research efforts focus on single pathogens, mixed infections are common in agriculture and can yield outcomes that cannot be predicted by studying pathogens in isolation (Barrett et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). In potato diseases, interactions between \u003cem\u003ePectobacterium\u003c/em\u003e and \u003cem\u003ePseudomonas\u003c/em\u003e species are particularly intricate, involving both antagonistic and pathogenic relationships (Terletskiy \u0026amp; Lazarev, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Aghdam et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Some \u003cem\u003ePseudomonas\u003c/em\u003e strains can inhibit \u003cem\u003ePectobacterium\u003c/em\u003e through biocontrol, whereas others promote disease, with environmental factors such as temperature playing a significant role (Yang et al., \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Alattas et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; H\u0026ouml;fte et al., 2021). Although evidence indicates both synergistic and antagonistic effects, the mechanisms underlying cooperation, competition, and increased pathogenicity among these bacteria remain poorly understood (Mahmoudi et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2011\u003c/span\u003e).\u003c/p\u003e \u003cp\u003ePrevious research has documented cases of synergistic disease enhancement. For example, studies have demonstrated that co-infection of plants with multiple viruses or with combinations of fungi and bacteria can increase disease severity (Bellah et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). However, the interactions between key potato pathogens, such as Pectobacterium species, and other common plant-associated bacteria remain poorly understood, particularly under varying environmental conditions. Temperature is a crucial abiotic factor that significantly affects both pathogen virulence and plant defense mechanisms. The optimal temperature for pathogen growth and infection varies widely across species, and temperature stress can weaken plant immunity, thereby increasing susceptibility to infection (Jee et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eTo resolve the knowledge gap, the present study explores how selected strains of \u003cem\u003ePectobacterium\u003c/em\u003e and \u003cem\u003ePseudomonas\u003c/em\u003e interact pathogenically with potatoes across various temperatures. By clarifying these interactions, the study provides a better understanding of pathogenic interactions between Pectobacterium spp. and Pseudomonas syringae in potato under co-inoculation and varying temperatures, thereby supporting the development of more effective and sustainable control strategies. The study involved two main hypotheses, including (i) co-inoculation of Pectobacterium strains with \u003cem\u003eP. syringae\u003c/em\u003e results in a synergistic increase in lesion development compared with each strain applied separately; and (ii) different temperature regimes influence disease progression in both individual and combined inoculations. The objectives of this study were: (i) to determine the pathogenicity of several \u003cem\u003ePectobacterium\u003c/em\u003e and \u003cem\u003eP. syringae\u003c/em\u003e strains on potato tubers; (ii) to determine whether co-inoculation results in synergistic or antagonistic effects on lesion size; and (iii) to assess the influence of different temperature regimes on disease progression in both individual and combined inoculations.\u003c/p\u003e"},{"header":"2. MATERIALS AND METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Experimental Facility, Material, Design, and Treatments\u003c/h2\u003e \u003cp\u003eThe study was conducted in the Department of Agrobiotechnology's controlled laboratory facility at RUDN University\u0026rsquo;s Institute of Agriculture in Russia to evaluate bacterial pathogenicity and interactions with potato tubers under varying temperature conditions. The experimental materials were tubers of the Gala potato variety. The experiment was conducted using a completely randomized design with four replicates. All inoculations were performed in complete darkness to prevent light from affecting bacterial growth or host responses. Two complementary experiments were designed. In the first, tubers were inoculated with individual bacterial strains, co-inoculated with each \u003cem\u003ePectobacterium\u003c/em\u003e strain and \u003cem\u003ePseudomonas syringae\u003c/em\u003e Ps3004, or treated with a sterile buffer control, then incubated at a constant 30\u0026deg;C to evaluate potential synergistic or antagonistic effects. The second experiment examined how temperature influences pathogenicity using selected strains (F148, F126, KC, Ps3004) inoculated individually and in combination (F148\u0026thinsp;+\u0026thinsp;Ps3004; F152\u0026thinsp;+\u0026thinsp;Ps3004; F126\u0026thinsp;+\u0026thinsp;Ps3004; and KC\u0026thinsp;+\u0026thinsp;Ps3004). The potato tubers were incubated at 5\u0026deg;C, 25\u0026deg;C, 30\u0026deg;C, and 35\u0026deg;C. A control group was kept at each temperature to ensure accurate comparison of disease responses.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Bacterial Inoculation and Colonization Experiment with Potato Tubers\u003c/h2\u003e \u003cp\u003eA total of four single-bacterial inoculation experiments were conducted using eight \u003cem\u003ePectobacterium\u003c/em\u003e strains and one \u003cem\u003ePseudomonas syringae\u003c/em\u003e Ps3004. The inoculation was performed according to the procedure described by K\u0026otilde;iv et al. (\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) and El-Hendawy et al. (2016). Accordingly, the strains were separately inoculated in liquid LB medium and cultured on suitable nutrient agar to produce enough biomass for the experiments. Cell suspensions were prepared in sterile buffer and adjusted to a uniform inoculum density by spectrophotometric measurement. The concentrations of the bacterial cultures were determined at an OD\u003csub\u003e600\u003c/sub\u003e of 0.5, and 100 \u0026micro;L of bacterial suspension was collected for each strain. Bacterial suspensions for each strain were prepared by mixing 0.4 mL of sterile water with 0.4 mL of bacterial culture. Potato tubers were surface-sterilized with NaClO (1%) for 5 min and thoroughly rinsed with sterile water three times and air-dried (Lee et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). The sterilized tubers were cut in half, and uniform wounds were created with a 15 mm cork borer (1 cm deep, 0.5 cm wide). The samples were sown in sterile plastic chambers lined with 2\u0026ndash;3 layers of moist filter paper towels to maintain humidity and incubated at 5\u0026deg;C, 25\u0026deg;C, 30\u0026deg;C, and 35\u0026deg;C. A control group was kept at each temperature to ensure accurate comparison of disease responses. Four healthy potato tubers were then placed in sterile chambers with 2\u0026ndash;3 layers of filter paper and were separately treated with 50 \u0026micro;L of bacterial culture suspension. The treated samples were kept at the various temperatures, and sterile water was added promptly to keep the filter paper moist until three days after inoculation. The control tubers were treated with a sterile medium solution. The experiment for each bacterial strain was conducted in four replicates. This method, adapted from K\u0026otilde;iv et al. (\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) and El-Hendawy et al. (2016), provided controlled, reproducible conditions for studying bacterial pathogenicity and interactions.\u003c/p\u003e \u003cp\u003eFor the co-bacterial inoculation experiment, equal volumes (0.4 mL) of each standardized suspension were mixed just before inoculation to ensure reliable bacterial interactions. The same standardized surface-sterilization protocol and inoculation technique described above were utilized. A total of 8 bacterial strains were mixed with \u003cem\u003ePseudomonas syringae\u003c/em\u003e Ps3004 to form the eight mixed bacterial complexes. Based on these 9 strains and the 8 mixed bacterial complexes, we compared the effects of single- and mixed-bacterial inoculation under diverse temperatures on the pathogenicity in potato. The experiment with 9 single-bacterial and 8 mixed-bacterial inoculations was performed using the same protocol as that described above.\u003c/p\u003e \u003cp\u003eFor temperature-dependent pathogenicity assays, selected treatments were additionally incubated at 5\u0026deg;C, 25\u0026deg;C, 30\u0026deg;C, and 35\u0026deg;C. Synergism experiments were mainly performed at 25\u0026deg;C or 30\u0026deg;C, which are optimal growth temperatures for these pathogens (Van der Wolf et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Du Raan et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Toth et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Inoculation involved creating a standardized wound with a 15 mm Cork borer, then pipetting 0.5 mL of either a single bacterial strain or a mixed bacterial suspension into each wound. Control treatments received an equal volume of sterile buffer. Each treatment was replicated four times, and the entire experiment was repeated four times. Tubers were kept in darkness to prevent light effects. After three days, tubers were sliced to evaluate rotting and disease incidence.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe \u003cem\u003ePectobacterium\u003c/em\u003e strains and \u003cem\u003ePseudomonas syringae\u003c/em\u003e Ps3004 used in this work\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003e#\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSpecies\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStrain designation\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eOrigin (Year of Isolation)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTaxonomic marker\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNCBI accession number\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePectobacterium brasiliense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eF126\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eSamara (2012)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003ecomplete genome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNZ_CP065031.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePectobacterium brasiliense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eF152\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMoscow (2014)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003ecomplete genome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNZ_PJDM00000000.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003ePectobacterium versatile\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eF131 (=\u0026thinsp;F137)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eKaluga (1998)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003ecomplete genome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNZ_PDVW01000000\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePectobacterium \u003cem\u003ecarotovorum\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eF164(PB31)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMoscow (2016)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003ecomplete genome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003ePJJA00000000.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003ePectobacterium parmentieri\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePb20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eSamara region (2012)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003ecomplete genome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNZ_PDDJ00000000\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003ePectobacterium parmentieri\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eF148 (PB20)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMoscow region (2013)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e16S rRNA, BOX-PCR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNZ_PDDJ00000000.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003ePectobacterium parmentieri\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eF034\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eKaluga\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eRecombinase A (recA) gene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMG518489.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003ePectobacterium\u003c/em\u003e sp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eKC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMoscow region (2025)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e16S rRNA, BOX-PCR\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eThis study\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003ePseudomonas syringae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePs3004\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eRostov region (2001)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e16S rRNA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eThis study\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Lesion Measurement\u003c/h2\u003e \u003cp\u003eDisease severity was assessed using a standard lesion-based scoring system used in potato soft rot studies (Lee et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Chung et al. 2017). After 72 h of incubation, tubers were sliced lengthwise through the inoculation point, and disease progression was evaluated by measuring lesion length and width (mm) with a digital caliper (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The lesion area was calculated as length \u0026times; width (mm\u0026sup2;) and served as the primary indicator of disease severity. This method provides a continuous, objective measure of tissue maceration and is widely used to evaluate the pathogenicity and virulence of \u003cem\u003ePectobacterium\u003c/em\u003e spp. and \u003cem\u003ePseudomonas syringae\u003c/em\u003e in potato tubers (Ge et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Czajkowski et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; K\u0026otilde;iv et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). For each treatment, lesion areas on individual tubers (n\u0026thinsp;=\u0026thinsp;4 per replicate) were measured, and the results were reported as mean values\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation. In co-inoculation experiments, microbial interactions were assessed by calculating the percentage change in lesion area compared to single-strain inoculations. Positive values signified synergistic effects, while negative values indicated antagonistic interactions. This standardized scoring system enabled reliable comparisons of disease severity across strains, treatments, and temperature conditions.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eData Collection and Analysis\u003c/b\u003e \u003c/p\u003e \u003cp\u003eAfter incubation, tubers were assessed for disease severity by measuring lesion size at the inoculation site. Each tuber was sliced through the center of the lesion (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), and both lesion length and width were recorded in millimeters. The lesion area, obtained by multiplying length and width, served as a standardized metric of disease severity in square millimeters. For each treatment group, the average lesion area was calculated from the individual replicates. The data were compiled, and mean values and standard deviations were determined to assess variability. Treatment differences were analyzed using mean lesion area and percentage change. Results were summarized using descriptive statistics to highlight patterns and significant differences. Statistical evaluation involved ANOVA and post hoc tests to compare strains and treatments at an α level of 0.05 (Ge et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003ePercentage Change Calculation: To determine the percentage change in lesion size (synergism and antagonism). We calculate the percentage increase or decrease in lesion area for co-inoculated treatments relative to the lesion area caused by the Pectobacterium strain alone using the formula:\u003c/p\u003e \u003cp\u003e\u003cimg 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\" width=\"475\" height=\"82\"\u003e\u003c/p\u003e\u003cp\u003eWhere Lesion Area\u003csub\u003e\u003cb\u003ems\u003c/b\u003e\u003c/sub\u003e= mean lesion area of mixed inoculation (co-inoculated) strain treatment and Lesion Area\u003csub\u003ess\u003c/sub\u003e = mean lesion area of single inoculation (single strain). Positive values indicate a synergistic effect (increased lesion size), whereas negative values indicate an antagonistic effect (reduced lesion size); see Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. Negative values indicate decreased lesion sizes due to Ps3004; positive values indicate increased lesion sizes, reflecting strain-specific responses.\u003c/p\u003e \u003c/div\u003e"},{"header":"3. RESULT","content":"\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e3.1 Pathogenicity of Single and Co-inoculation Bacterial Strains\u003c/h2\u003e \u003cp\u003eThe average lesion area (mm\u0026sup2;) varied considerably among the SRP strains when inoculated alone and in combination with Pseudomonas syringae Ps3004. When applied individually, the virulence of each bacterial strain varied significantly. The results, summarized in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, indicate that not all strains were pathogenic under the experimental conditions. Pathogenic strains included \u003cem\u003eP. parmentieri\u003c/em\u003e (Pb20, F034, F148), \u003cem\u003eP. brasiliense\u003c/em\u003e (F152, F126), and \u003cem\u003eP. syringae\u003c/em\u003e (Ps3004), all of which caused visible lesions on potato tubers. Lesion sizes ranged from an average of 103 mm\u003csup\u003e2\u003c/sup\u003e for Ps3004 to 316 mm\u003csup\u003e2\u003c/sup\u003e for \u003cem\u003eP. brasiliense\u003c/em\u003e (F126). Non-pathogenic strains involved \u003cem\u003eP. carotovorum\u003c/em\u003e (F164), which did not produce any visible lesions; \u003cem\u003eP. versatile\u003c/em\u003e strain F131, and \u003cem\u003ePectobacterium\u003c/em\u003e sp. (KC), which failed to produce lesions at +\u0026thinsp;50\u0026deg;C and +\u0026thinsp;25\u0026deg;C; and \u003cem\u003ePectobacterium\u003c/em\u003e sp. (KC), which did not produce lesions at +\u0026thinsp;50\u0026deg;C. The control (K) showed no reaction, confirming that the lesions were caused by bacterial activity.\u003c/p\u003e \u003cp\u003eCo-inoculating weakly pathogenic \u003cem\u003ePectobacterium\u003c/em\u003e strains with \u003cem\u003eP. syringae\u003c/em\u003e Ps3004 significantly increased lesion severity, especially with \u003cem\u003eP. carotovorum\u003c/em\u003e (F164), \u003cem\u003eP. parmentieri\u003c/em\u003e (F148), \u003cem\u003eP. versatile\u003c/em\u003e strain F131, and \u003cem\u003ePectobacterium\u003c/em\u003e sp. (KC). A moderate synergistic effect was observed with \u003cem\u003ethe P. versatile\u003c/em\u003e strain F131, which increased lesion area by 94.4% when combined with Ps3004. The combination of F148 and Ps3004 caused a 9.6% increase, whereas \u003cem\u003ePectobacterium\u003c/em\u003e sp (KC) increased by 7.4%, and \u003cem\u003eP. carotovorum\u003c/em\u003e F164 exhibited a substantial increase of 4300%.\u003c/p\u003e \u003cp\u003eConversely, co-inoculating highly virulent \u003cem\u003ePectobacterium\u003c/em\u003e strains with \u003cem\u003eP. syringae Ps3004\u003c/em\u003e reduced lesion severity, demonstrating an antagonistic effect. This was particularly evident in \u003cem\u003eP. parmentieri\u003c/em\u003e (Pb20, F034) and \u003cem\u003eP. brasiliense\u003c/em\u003e (F126). For example, combining Pb20 with Ps3004 resulted in an average lesion area of 432 mm\u003csup\u003e2\u003c/sup\u003e, a 32.2% reduction compared with Pb20 alone. The pairing of F034 and Ps3004 resulted in a lesion area of 448 mm\u003csup\u003e2\u003c/sup\u003e, a 43.8% decrease over F034 alone. Figure\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e displays soft rot symptoms in potato tubers inoculated with different bacterial strains: \u003cem\u003ePseudomonas syringae\u003c/em\u003e Ps3004 (a), \u003cem\u003ePectobacterium\u003c/em\u003e sp. KC (b), \u003cem\u003ePectobacterium brasiliense\u003c/em\u003e F148 (c), and \u003cem\u003ePectobacterium brasiliense\u003c/em\u003e F126 (d). The tubers were kept in darkness at 30\u0026deg;C for 48 h. The images highlight the strain-dependent variations in tissue maceration, discoloration, and exudate formation.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eEffects of co-inoculation on lesion size caused by pectotytic bacteria. (A) Shows the average lesion area (mm\u003csup\u003e2\u003c/sup\u003e) after co-inoculation with Ps3004. (B) Displays the average squared lesion area (mm\u003csup\u003e2\u003c/sup\u003e) for single-strain inoculation. The percentage change indicates the change in lesion area when Ps3004 is combined with the strain relative to the strain alone.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStrains\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSpecies\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAverage squared lesion area (mm\u003csup\u003e2\u003c/sup\u003e) + Ps3004 (A)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eAv. squared lesion area (mm\u003csup\u003e2\u003c/sup\u003e) (B)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003ePercentage change (%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePb20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eP. parmentieri\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e293\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e432\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-139(-32.2%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF034\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eP. parmentieri\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e252\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e448\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-196(-43.8%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF148\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eP. parmentieri\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e286\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e261\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u0026thinsp;25(9.6%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF152\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eP. brasiliense\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e308\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e353\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-45(-12.7%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF164\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eP. carotovorum\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u0026thinsp;64.5(4300%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF126\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eP. brasiliense\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e316\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e357\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-41(-11.5%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF131\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eP. versatile\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e241\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e124\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u0026thinsp;117(94.4%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eKC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003ePectobacterium\u003c/em\u003e sp\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e246\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e229\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u0026thinsp;17(7.4%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e3.2 Influence of Temperature on Pathogenicity\u003c/h2\u003e \u003cp\u003eTemperature played a crucial role in lesion development and the interactions among the tested strains, emphasizing its importance in controlling soft rot severity. As indicated in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e, no lesions formed at 5\u0026deg;C, showing that bacterial activity was suppressed at low temperatures. Conversely, disease severity increased significantly at 25\u0026deg;C and peaked at 30\u0026deg;C for the most aggressive strains, particularly Pectobacterium brasiliense F126 and P. parmentieri F148. The decrease in lesion size at 35\u0026deg;C points to thermal stress or a decline in virulence-related processes at temperatures above the optimum.\u003c/p\u003e \u003cp\u003eCo-inoculation changed how temperature affects pathogen responses. Combining P. brasiliense F126 with Pseudomonas syringae Ps3004 extended the temperature range over which pathogenic effects are observed, maintaining visible lesions from 25\u0026deg;C to 35\u0026deg;C (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Similarly, Pectobacterium sp. KC showed increased lesion formation at intermediate temperatures when co-inoculated, despite having limited virulence on its own. These results imply that interspecies interactions can help buffer against environmental stress and alter virulence levels. The temperature-dependent patterns align with earlier research showing that enzyme activity, motility, and quorum sensing in Soft Rot Pectobacteriaceae are most active at moderate temperatures and decrease under extreme heat or cold (Czajkowski et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Jee et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Toth et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eTemperature-dependent pathogenicity of selected bacterial strains and co-inoculations on potato tubers, highlighting the synergistic and antagonistic effects of co-inoculation with \u003cem\u003ePseudomonas syringae\u003c/em\u003e Ps3004, \u003cem\u003eP. brasiliense\u003c/em\u003e (F126, F152), \u003cem\u003eP. versatile\u003c/em\u003e strain F131, \u003cem\u003eP\u003c/em\u003e. \u003cem\u003eparmentieri\u003c/em\u003e (Pb20, F034, F148), \u003cem\u003eP. carotovorum\u003c/em\u003e (F164), and \u003cem\u003ePectobacterium\u003c/em\u003e sp. (KC), with \u003cem\u003ePseudomonas syringae\u003c/em\u003e Ps3004\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cem\u003ePseudomonas syringae\u003c/em\u003e (Ps3004)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cem\u003eP. Brasiliense\u003c/em\u003e (F152)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cem\u003eP. parmentier\u003c/em\u003e (F148)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cem\u003ePectobacterium\u003c/em\u003e sp. KC\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c8\" namest=\"c5\"\u003e \u003cp\u003eMean scores (mm\u0026sup2;)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5\u0026deg;C\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e25\u0026deg;C\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e30\u0026deg;C\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e35\u0026deg;C\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e272\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e270\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e375\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e198\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e308\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e480\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e972\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e779\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e210\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e462\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e306\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e180\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e273\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e460\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e295\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e268\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e338\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e476\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e224\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"8\"\u003eSymbols (+) and (\u0026ndash;) indicate the presence and absence of a bacterial strain during inoculation, respectively\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eLesion development (mm\u0026sup2;) of potato tubers inoculated with selected bacterial strains and their combinations under different temperature regimes (5, 25, 30, and 35\u0026deg;C) (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Values are presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD (n\u0026thinsp;=\u0026thinsp;4). For each temperature, treatment means were compared using one-way ANOVA; means that differed by lowercase letter within the same temperature column were considered significantly different based on post hoc multiple comparisons (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e \u003c/div\u003e"},{"header":"4. DISCUSSION","content":"\u003cp\u003eThis study presents experimental evidence that disease severity in potato tubers is influenced by bacterial strain identity, interspecific interactions, and temperature. Notable differences in virulence were found among Soft Rot \u003cem\u003ePectobacterium\u003c/em\u003e bacteria and \u003cem\u003ePseudomonas syringae\u003c/em\u003e, underscoring the complexity of mixed bacterial infections. \u003cem\u003ePectobacterium parmentieri\u003c/em\u003e (Pb20, F034, F148), \u003cem\u003eP. brasiliense\u003c/em\u003e (F126, F152), and \u003cem\u003eP. syringae\u003c/em\u003e (Ps3004) were pathogenic when tested individually, while \u003cem\u003eP. carotovorum\u003c/em\u003e (F164) was non-pathogenic under the experimental conditions, suggesting strain-specific or environment-dependent virulence. The lack of lesions in control treatments confirms that lesion formation was caused solely by bacterial activity.\u003c/p\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e4.1 Pathogenic Variability among SRP Strains\u003c/h2\u003e \u003cp\u003eThe present study revealed pronounced pathogenic variability among Soft Rot Pectobacteriaceae (SRP) strains, confirming that virulence within this group is highly strain dependent and cannot be inferred solely from species identity. When inoculated individually, \u003cem\u003ePectobacterium parmentieri\u003c/em\u003e (Pb20, F034, F148) and \u003cem\u003eP. brasiliense\u003c/em\u003e (F126, F152) consistently induced large lesions on potato tubers, whereas \u003cem\u003eP. carotovorum\u003c/em\u003e (F164), \u003cem\u003eP. versatile\u003c/em\u003e (F131), and \u003cem\u003ePectobacterium\u003c/em\u003e sp. (KC) exhibited weak or negligible pathogenicity under the same experimental conditions. These findings clearly demonstrate substantial intra- and interspecific heterogeneity in pathogenic potential among SRP strains (Czajkowski et al. 2011; Toth et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Such variability is well documented in SRP pathosystems and reflects differences in pectinolytic enzyme production, quorum-sensing regulation, stress tolerance, and environmental responsiveness rather than taxonomic classification alone (Czajkowski et al. 2011; Toth et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Previous studies have shown that isolates of \u003cem\u003eP. brasiliense\u003c/em\u003e and \u003cem\u003eP. parmentieri\u003c/em\u003e differ markedly in aggressiveness, maceration kinetics, and temperature optima, even when recovered from the same host or geographic region (Golanowska et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; van der Wolf et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Our results are consistent with these reports, as strains within the same species exhibited contrasting pathogenic behaviors.\u003c/p\u003e \u003cp\u003eThe absence of lesion formation by \u003cem\u003eP. carotovorum\u003c/em\u003e F164 and \u003cem\u003ePectobacterium\u003c/em\u003e sp. KC during individual inoculation indicates that some SRP strains can survive in potato tissues as low-virulence or opportunistic colonizers. Their ability to cause disease appears to depend on specific environmental conditions or the presence of other microbes. Similar findings have been reported for SRP isolates with reduced pectate lyase activity or compromised virulence gene expression, which induce disease only when additional stressors or interacting microorganisms are present (Davidsson et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Czajkowski et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Overall, the variability in pathogenic traits underscores the limitations of single-strain or species-level models for predicting the risk of potato soft rot. Disease outcomes are influenced by a mix of strain-specific characteristics, environmental factors, and microbial interactions. Understanding this diversity is essential for improving disease diagnostics, refining risk assessment models, and developing targeted management strategies that account for the ecological complexity of SRP populations in potato production systems.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e4.2 Synergistic and Antagonistic Interactions with \u003cem\u003ePseudomonas syringae\u003c/em\u003e\u003c/h2\u003e \u003cp\u003eCo-inoculation experiments revealed that interactions between SRP strains and \u003cem\u003eP. syringae\u003c/em\u003e Ps3004 vary and are not always mutually beneficial. Some combinations led to more severe lesions, while others reduced disease severity, indicating antagonistic effects (Fig.\u0026nbsp;3). The most pronounced increases were observed with strains that were weakly pathogenic or non-pathogenic alone, such as \u003cem\u003eP. brasiliense\u003c/em\u003e F137, \u003cem\u003eP\u003c/em\u003e. sp. KC, and \u003cem\u003eP. carotovorum\u003c/em\u003e F164. These results align with previous studies indicating that mixed bacterial infections can amplify disease severity through mechanisms such as tissue maceration, changes in quorum sensing, or suppression of host defenses (Degefu et al. 2021).\u003c/p\u003e \u003cp\u003eIn contrast, the antagonistic effects observed with \u003cem\u003eP. parmentieri\u003c/em\u003e (Pb20, F034) and \u003cem\u003eP. brasiliense\u003c/em\u003e (F126) suggest competitive exclusion or interference among coexisting bacteria. Similar results have been reported in SRP pathosystems, where competition for nutrients, space, or oxygen can reduce virulence in mixed infections (Arif et al. 2022). These responses indicate that mixed infections do not necessarily lead to more severe disease, as outcomes depend on strain-specific traits and ecological factors.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e4.3 Temperature as a Modulator of Pathogenicity and Interaction Outcomes\u003c/h2\u003e \u003cp\u003eTemperature significantly influenced disease expression, reaffirming its role as a key environmental factor affecting SRP virulence. The highest lesion development for \u003cem\u003eP. syringae\u003c/em\u003e Ps3004 and \u003cem\u003eP. brasiliense\u003c/em\u003e F126 occurred at 30\u0026deg;C, with a decline at 35\u0026deg;C. The combination of \u003cem\u003eP. brasiliense\u003c/em\u003e F126 and \u003cem\u003eP. syringae\u003c/em\u003e Ps3004 caused disease over a wider temperature range (25\u0026deg;C to 35\u0026deg;C), with lesion severity increasing with temperature. Similarly, \u003cem\u003eP. parmentieri\u003c/em\u003e F148 caused disease across this range when inoculated alone, exhibiting high pathogenicity at 30\u0026deg;C and reduced pathogenicity at 35\u0026deg;C. The strain KC (\u003cem\u003ePectobacterium\u003c/em\u003e sp) showed linear disease symptoms only between 25\u0026deg;C and 35\u0026deg;C when co-inoculated with \u003cem\u003eP. syringae\u003c/em\u003e (Ps3004). These temperature-dependent behaviors illustrate the complex interaction between environmental conditions and pathogen activity in disease development.\u003c/p\u003e \u003cp\u003eThis aligns with earlier research indicating that SRP virulence factors, including pectolytic enzyme activity and motility, are most effective at moderate temperatures and decrease under thermal stress (Czajkowski et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Jee et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Toth et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). For \u003cem\u003eP. brasiliense\u003c/em\u003e (F126), pectolytic activity of \u003cem\u003ePectobacterium\u003c/em\u003e diminished at 35\u0026deg;C when co-inoculated with \u003cem\u003eP. syringae\u003c/em\u003e (Ps3004), in contrast to \u003cem\u003eP. parmentieri\u003c/em\u003e (F148).\u003c/p\u003e \u003cp\u003eNotably, co-inoculating \u003cem\u003eP. brasiliense\u003c/em\u003e F126 with Ps3004 broadened the temperature range for disease development, demonstrating that interspecific interactions can mitigate environmental limits. Similar temperature-dependent effects on disease severity have been observed in mixed infections involving \u003cem\u003eDickeya\u003c/em\u003e and \u003cem\u003ePectobacterium\u003c/em\u003e species, where cooperative interactions intensify under favorable climatic conditions (van der Wolf et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Degefu, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Jee et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Lebecka, et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). These results are especially significant given predictions of climate change, which suggest an increase in conditions conducive to SRP outbreaks.\u003c/p\u003e \u003c/div\u003e"},{"header":"5. CONCLUSION","content":"\u003cp\u003eThis study demonstrated that bacterial strain, interspecific interactions from co-inoculation involving \u003cem\u003ePectobacterium\u003c/em\u003e spp. and \u003cem\u003ePseudomonas syringae\u003c/em\u003e, and temperature influence disease development (pathogenicity and disease severity) in potato tubers, findings that could be exploited to develop more effective and sustainable control strategies and to improve potato breeding for disease resistance. The combined evidence from pathogenicity, interaction, and temperature experiments indicates that relying solely on single-pathogen models for disease risk assessments might underestimate the complexity of SRP epidemics. Mixed infections can either worsen or suppress disease outcomes depending on strain combinations and environmental factors. Enhancing detection of latent infections and understanding interaction-driven disease dynamics will be crucial for sustainable management.\u003c/p\u003e \u003cp\u003ePathogenicity varied significantly among SRP strains, reflecting substantial intra-species heterogeneity. Co-inoculation with \u003cem\u003ePseudomonas syringae\u003c/em\u003e Ps3004 yielded both synergistic and antagonistic effects, demonstrating that mixed infections do not always lead to increased disease severity. Temperature heavily influenced pathogenic outcomes, with peak virulence typically at 30\u0026deg;C and diminished disease expression at higher temperatures.\u003c/p\u003e \u003cp\u003eFindings suggest that SRP epidemics are driven by strain-specific traits, microbial interactions, and environmental factors rather than a single cause. These findings support the importance of integrated disease management strategies that account for ecological complexity, especially amid changing climate conditions. Future research should explore the mechanisms underlying interaction-driven virulence, improve early detection of latent infections, and develop predictive models that incorporate microbial interactions and environmental influences.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors acknowledge the support given bytheDepartment of Agrobiotechnology, Agrarian-Technological Institute, RUDN University, 117198 Moscow, Russia.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors agree with the submission of this manuscript to Plant Methods.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe research was supported by the Russian Ministry of Education and Science Project FSSF-2024-0063.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability Statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data that support the findings of this study are available from the corresponding author upon reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors have declaredno known conflicts of interest at the time of writing this article. The authors affirm that they have no recognized competing financial interests or personal connections that could have appeared to affect the work presented in this paper.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAghdam, N. M. N., Baghaee-Ravari, S., \u0026amp; Shiri, A. (2023). Antimicrobial capacity of Pseudomonas brassicacearum strain EnPb against potato soft rot agent. European Journal of Plant Pathology, 165(2), 215\u0026ndash;231. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10658-022-02600-z\u003c/span\u003e\u003cspan address=\"10.1007/s10658-022-02600-z\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAlattas, H., Glick, B. R., Murphy, D. V., \u0026amp; Scott, C. (2024). Harnessing Pseudomonas spp. for sustainable plant crop protection. Frontiers in microbiology, 15, 1485197. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fmicb.2024.1485197\u003c/span\u003e\u003cspan address=\"10.3389/fmicb.2024.1485197\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eArif, M., Busot, G. Y., Mann, R., Rodoni, B., \u0026amp; Stack, J. P. (2021). Field-deployable recombinase polymerase amplification assay for specific, sensitive and rapid detection of the US select agent and toxigenic bacterium, Rathayibacter toxicus. Biology, 10(7), 620. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/biology10070620\u003c/span\u003e\u003cspan address=\"10.3390/biology10070620\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBabinska-Wensierska, W., Motyka-Pomagruk, A., Fondi, M., Misztak, A. E., Mengoni, A., \u0026amp; Lojkowska, E. (2024). Differences in the constituents of bacterial microbiota of soils collected from two fields of diverse potato blackleg and soft rot diseases incidences, a case study. Scientific Reports, 14(1), 18802. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/s41598-024-69213-w\u003c/span\u003e\u003cspan address=\"10.1038/s41598-024-69213-w\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBarrett, L. G., Zala, M., Mikaberidze, A., Alassimone, J., Ahmad, M., McDonald, B. A., \u0026amp; S\u0026aacute;nchez-Vallet, A. (2021). Mixed infections alter transmission potential in a fungal plant pathogen. Environmental microbiology, 23(4), 2315\u0026ndash;2330. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/1462-2920.15417\u003c/span\u003e\u003cspan address=\"10.1111/1462-2920.15417\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBellah, H., Seiler, N. F., \u0026amp; Croll, D. (2023). Divergent outcomes of direct conspecific pathogen strain interaction and plant co-infection suggest consequences for disease dynamics. Microbiology Spectrum, 11(2), e04443-22. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1128/spectrum.04443-22\u003c/span\u003e\u003cspan address=\"10.1128/spectrum.04443-22\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChen, H., Chen, J., Zhao, Y., Liu, F., \u0026amp; Fu, Z. Q. (2022). Pseudomonas syringae pathovars. Trends in Microbiology, 30(9). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.tim.2022.03.002\u003c/span\u003e\u003cspan address=\"10.1016/j.tim.2022.03.002\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChung YongSuk, C. Y., Kim ChangSoo, K. C., \u0026amp; Jansky, S. (2017). New source of bacterial soft rot resistance in wild potato (Solanum chacoense) tubers. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10722-017-0487-3\u003c/span\u003e\u003cspan address=\"10.1007/s10722-017-0487-3\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCzajkowski, R. (2011). Ecology and control of Dickeya spp. in potato. Netherlands Institute of Ecology. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://hdl.handle.net/1887/17810\u003c/span\u003e\u003cspan address=\"https://hdl.handle.net/1887/17810\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCzajkowski, R., Kaczyńska, N., Jafra, S., Narajczyk, M., \u0026amp; Lojkowska, E. (2017). Temperature-responsive genetic loci in pectinolytic plant pathogenic Dickeya solani. Plant Pathology, 66(4), 584\u0026ndash;594. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/ppa.12618\u003c/span\u003e\u003cspan address=\"10.1111/ppa.12618\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDavidsson, P. R., Kariola, T., Niemi, O., \u0026amp; Palva, E. T. (2013). Pathogenicity of and plant immunity to soft rot pectobacteria. Frontiers in plant science, 4, 191. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fpls.2013.00191\u003c/span\u003e\u003cspan address=\"10.3389/fpls.2013.00191\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDegefu, Y., 2021. Co-occurrence of latent Dickeya and Pectobacterium species in potato seed tuber samples from northern Finland: Co-colonization of latent Dickeya and Pectobacterium species in potato seed lots. Agricultural and Food Science, 30(1), pp.1\u0026ndash;7. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.23986/afsci.101446\u003c/span\u003e\u003cspan address=\"10.23986/afsci.101446\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDegefu, Y., 2024. Lesson from the emergence, spread and decline of Dickeya solani, the virulent potato blackleg and soft rot bacterial pathogen in Finland. Journal of phytopathology, 172(2), p.e13282. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/jph.13282\u003c/span\u003e\u003cspan address=\"10.1111/jph.13282\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDu Raan, S., Coutinho, T. A., \u0026amp; Van der Waals, J. E. (2016). Cardinal temperature differences, determined in vitro, between closely related species and subspecies of pectinolytic bacteria responsible for blackleg and soft rot on potatoes. European Journal of Plant Pathology, 144(2), 361\u0026ndash;369. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10658-015-0773-x\u003c/span\u003e\u003cspan address=\"10.1007/s10658-015-0773-x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDupuis, B., Nkuriyingoma, P., \u0026amp; Van Gijsegem, F. (2021). Economic impact of Pectobacterium and Dickeya species on potato crops: A review and case study. In Plant diseases caused by dickeya and pectobacterium species (pp. 263\u0026ndash;282). Cham: Springer International Publishing. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/978-3-030-61459-1_8\u003c/span\u003e\u003cspan address=\"10.1007/978-3-030-61459-1_8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEl-Hendawy, H. H., \u0026amp; Abo-Elyousr, K. A. M. (2016). Combination of different antagonistic bacteria to control of potato blackleg disease caused by Pectobacterium atrosepticum under greenhouse and field conditions.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGe, T., Ekbataniamiri, F., Johnson, S. B., Larkin, R. P., \u0026amp; Hao, J. (2021). Interaction between Dickeya dianthicola and Pectobacterium parmentieri in potato infection under field conditions. Microorganisms, 9(2), 316. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/microorganisms9020316\u003c/span\u003e\u003cspan address=\"10.3390/microorganisms9020316\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGolanowska, M., Kielar, J., \u0026amp; Lojkowska, E. (2017). The effect of temperature on the phenotypic features and the maceration ability of Dickeya solani strains isolated in Finland, Israel and Poland. European Journal of Plant Pathology, 147, 803\u0026ndash;817. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10658-016-1044-1\u003c/span\u003e\u003cspan address=\"10.1007/s10658-016-1044-1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eH\u0026ouml;fte, M. (2021). The use of Pseudomonas spp. as bacterial biocontrol agents to control plant disease. In Microbial bioprotectants for plant disease management. Burleigh Dodds. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://dx.doi.org/10.19103/AS.2021.0093.11\u003c/span\u003e\u003cspan address=\"10.19103/AS.2021.0093.11\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHugouvieux-Cotte‐Pattat, N., Condemine, G., \u0026amp; Shevchik, V. E. (2014). Bacterial pectate lyases, structural and functional diversity. Environmental microbiology reports, 6(5), 427\u0026ndash;440. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/1758-2229.12166\u003c/span\u003e\u003cspan address=\"10.1111/1758-2229.12166\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJee, S., Choi, J. G., Lee, Y. G., Kwon, M., Hwang, I., \u0026amp; Heu, S. (2020). Distribution of Pectobacterium species isolated in South Korea and comparison of temperature effects on pathogenicity. The plant pathology journal, 36(4), 346. https://orcid.org/0000-0001-7834-4303\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKatagiri, F., Thilmony, R., \u0026amp; He, S. Y. (2002). The Arabidopsis thaliana-Pseudomonas syringae interaction. The Arabidopsis Book/American Society of Plant Biologists, 1, e0039. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1199/tab.0039\u003c/span\u003e\u003cspan address=\"10.1199/tab.0039\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eK\u0026otilde;iv, V., Roosaare, M., Vedler, E., Ann Kivistik, P., Toppi, K., Schryer, D. W., \u0026hellip; M\u0026auml;e, A. (2015). Microbial population dynamics in response to Pectobacterium atrosepticum infection in potato tubers. Scientific reports, 5(1), 11606. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/srep11606\u003c/span\u003e\u003cspan address=\"10.1038/srep11606\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eK\u0026otilde;iv, V., Roosaare, M., Vedler, E., Ann Kivistik, P., Toppi, K., Schryer, D. W., \u0026hellip; M\u0026auml;e, A. (2015). Microbial population dynamics in response to Pectobacterium atrosepticum infection in potato tubers. Scientific reports, 5(1), 11606. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/srep11606\u003c/span\u003e\u003cspan address=\"10.1038/srep11606\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLebecka, R., Flis, B., \u0026amp; Murawska, Z. (2018). Comparison of temperature effects on the in vitro growth and disease development in potato tubers inoculated with bacteria Pectobacterium atrosepticum, P carotovorum subsp. carotovorum and Dickeya solani. Journal of Phytopathology, 166(9), 654\u0026ndash;662. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/jph.12728\u003c/span\u003e\u003cspan address=\"10.1111/jph.12728\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLee, U., Silva, R. R., Kim, C., Kim, H., Heo, S., Park, I. S., \u0026hellip; Chung, Y. S. (2019). Image analysis for measuring disease symptom to bacterial soft rot in potato. American Journal of Potato Research, 96(3), 303\u0026ndash;313. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s12230-019-09717-8\u003c/span\u003e\u003cspan address=\"10.1007/s12230-019-09717-8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMahmoudi, E., Ahmadi, A., Sayed-Tabatabaei, B. E., Ghobadi, C., Akhavan, A., Hasanzadeh, N., \u0026amp; Venturi, V. (2011). A novel AHL-degrading rhizobacterium quenches the virulence of Pectobacterium atrosepticum on potato plant. Journal of Plant Pathology, 587\u0026ndash;594. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://dx.doi.org/10.4454/jpp.v93i3.1226\u003c/span\u003e\u003cspan address=\"10.4454/jpp.v93i3.1226\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMansfield, J., Genin, S., Magori, S., Citovsky, V., Sriariyanum, M., Ronald, P., \u0026hellip; Foster, G. D. (2012). Top 10 plant pathogenic bacteria in molecular plant pathology. Molecular plant pathology, 13(6), 614\u0026ndash;629. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/j.1364-3703.2012.00804.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1364-3703.2012.00804.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOulghazi, S., Moumni, M., Khayi, S., Robic, K., Sarfraz, S., Lopez-Roques, C., Vandecasteele, C., \u0026amp; Faure, D. (2020). Diversity of Pectobacteriaceae Species in Potato Growing Regions in Northern Morocco. Microorganisms, 8(6), 895. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/microorganisms8060895\u003c/span\u003e\u003cspan address=\"10.3390/microorganisms8060895\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePortier, P., P\u0026eacute;dron, J., Taghouti, G., Dutrieux, C., \u0026amp; Barny, M. A. (2020). Updated taxonomy of Pectobacterium genus in the CIRM-CFBP bacterial collection: when newly described species reveal \u0026ldquo;old\u0026rdquo; endemic population. Microorganisms, 8(9), 1441. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/microorganisms8091441\u003c/span\u003e\u003cspan address=\"10.3390/microorganisms8091441\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSu, Z., Liu, X., Guo, Q., Xuan, L., Lu, X., Dong, L., \u0026hellip; \u0026amp; Ma, P. (2022). Insights into complex infection by two Pectobacterium species causing potato blackleg and soft rot. Microbiological Research, 261, 127072. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.micres.2022.127072\u003c/span\u003e\u003cspan address=\"10.1016/j.micres.2022.127072\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSzulta, S., \u0026amp; Kornicka, A. (2023). Pectobacterium and Dickeya genus\u0026mdash;A review on structural variations of O-polysaccharides and their role in the pathogenic process of plants. Plant Pathology, 72(6), 998\u0026ndash;1010. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/ppa.13728\u003c/span\u003e\u003cspan address=\"10.1111/ppa.13728\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTerletskiy, V. P., \u0026amp; Lazarev, A. M. (2019). On genotyping bacterial strains of the genera Pectobacterium and Pseudomonas: pathogens of bacterioses in potatoes. Cytology and Genetics, 53(3), 212\u0026ndash;218. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3103/S0095452719030058\u003c/span\u003e\u003cspan address=\"10.3103/S0095452719030058\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eToth, I. K., Barny, M. A., Brurberg, M. B., Condemine, G., Czajkowski, R., Elphinstone, J. G., \u0026hellip; Yedidia, I. (2021). Pectobacterium and Dickeya: environment to disease development. In Plant diseases caused by Dickeya and Pectobacterium species (pp. 39\u0026ndash;84). Cham: Springer International Publishing. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/978-3-030-61459-1_3\u003c/span\u003e\u003cspan address=\"10.1007/978-3-030-61459-1_3\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003evan der Wolf, J. M., Acu\u0026ntilde;a, I., De Boer, S. H., Brurberg, M. B., Cahill, G., Charkowski, A. O., \u0026hellip; Yedidia, I. (2021). Diseases caused by Pectobacterium and Dickeya species around the world. In Plant diseases caused by Dickeya and Pectobacterium species (pp. 215\u0026ndash;261). Cham: Springer International Publishing. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/978-3-030-61459-1_7\u003c/span\u003e\u003cspan address=\"10.1007/978-3-030-61459-1_7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVan der Wolf, J. M., De Haan, E. G., Kastelein, P., Krijger, M., De Haas, B. H., Velvis, H., \u0026hellip; Van Der Zouwen, P. S. (2017). Virulence of Pectobacterium carotovorum subsp. brasiliense on potato compared with that of other Pectobacterium and Dickeya species under climatic conditions prevailing in the Netherlands. Plant Pathology, 66(4), 571\u0026ndash;583. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/ppa.12600\u003c/span\u003e\u003cspan address=\"10.1111/ppa.12600\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang, T., Hua, C., \u0026amp; Deng, X. (2023). c-di-GMP signaling in Pseudomonas syringae complex. Microbiological Research, 275, 127445. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.micres.2023.127445\u003c/span\u003e\u003cspan address=\"10.1016/j.micres.2023.127445\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYang, Z., Liu, T., Fan, J., Chen, Y., Wu, S., Li, J., \u0026hellip; Tang, Q. (2024). Biocontrol agents modulate phyllosphere microbiota interactions against pathogen Pseudomonas syringae. Environmental Science and Ecotechnology, 21, 100431. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.ese.2024.100431\u003c/span\u003e\u003cspan address=\"10.1016/j.ese.2024.100431\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhou, J., Hu, M., \u0026amp; Zhang, L. (2024). Dickeya Diversity and Pathogenic Mechanisms. Annual Review of Microbiology, 78(1), 621\u0026ndash;642. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1146/annurev-micro-041222-012242\u003c/span\u003e\u003cspan address=\"10.1146/annurev-micro-041222-012242\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Soft rot Pectobacteriacea, Polymicrobial infection, synergistic and antagonistic interactions, lesion area, tissue maceration, Solanum tuberosum","lastPublishedDoi":"10.21203/rs.3.rs-8833264/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8833264/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003ePotato production is challenged by bacterial soft rot and blackleg diseases caused by the Soft Rot Pectobacteriaceae (SRP), particularly Pectobacterium species. While \u003cem\u003ePseudomonas syringae\u003c/em\u003e, a common potato-related bacterium and well-studied plant pathogen, is known, its role in mixed infections with SRP under various environmental conditions remains unclear. This research examined the pathogenic potential of several Pectobacterium strains and \u003cem\u003eP. syringae\u003c/em\u003e on potato tubers, explored how different strains interact during co-infection either synergistically or antagonistically, and analyzed how temperature influences disease development. Potato tubers were inoculated with either a single strain or a combination of \u003cem\u003eP. syringae\u003c/em\u003e strains and then incubated at 5, 25, 30, and 35\u0026deg;C in controlled-environment chambers. Disease severity was measured by lesion area (mm\u0026sup2;), and interaction outcomes were expressed as percentage changes relative to single-strain infections. Significant differences in pathogenicity were observed among SRP strains; Pectobacterium parmentieri and \u003cem\u003eP. brasiliense\u003c/em\u003e exhibited high virulence, whereas other strains were weak or nonpathogenic when tested alone. Co-inoculation with \u003cem\u003eP. syringae\u003c/em\u003e yielded strain-specific effects: it heightened disease severity in weakly pathogenic strains but decreased lesion formation in highly virulent ones, suggesting antagonism. Temperature markedly affected disease expression, with maximum virulence usually at 30\u0026deg;C. Co-inoculation also extended the temperature range conducive to disease development. These results illustrate that potato soft rot severity depends on bacterial strain type, microbial interactions, and temperature, underscoring the limitations of single-pathogen models and highlighting the importance of considering microbial communities and environmental factors in disease prediction and control.\u003c/p\u003e","manuscriptTitle":"Pathogenic Interactions Between Pectobacterium spp. and Pseudomonas syringae in Potato as Influenced by Co-Inoculation and Temperature","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-02-20 09:41:25","doi":"10.21203/rs.3.rs-8833264/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"476e42c9-44b7-40ff-90ec-50b4b624c550","owner":[],"postedDate":"February 20th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-03-03T09:56:20+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-20 09:41:25","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8833264","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8833264","identity":"rs-8833264","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}