Biocontrol of Meloidogyne incognita and Vegetative Growth Stimulation in Tomato ‘Moneymaker’ Plants by Egyptian Soil Bacteria

Biocontrol of Meloidogyne incognita and Vegetative Growth Stimulation in Tomato ‘Moneymaker’ Plants by Egyptian Soil Bacteria | 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 Biocontrol of Meloidogyne incognita and Vegetative Growth Stimulation in Tomato ‘Moneymaker’ Plants by Egyptian Soil Bacteria Ahmed MA Mahmoud, Ahmed ASA El-Eslamboly, Mohamed Adam, Mahmoud AA Maraey This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6043078/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 13 Jun, 2025 Read the published version in Egyptian Journal of Biological Pest Control → Version 1 posted 6 You are reading this latest preprint version Abstract Background: Tomato yield is significantly reduced by root-knot nematodes (RKN; Meloidogyne spp.), particularly in tropical and subtropical regions. This study evaluated 20 bacterial isolates (B1-B20), belonging to the genera Bacillus , Lysobacter , Paenibacillus , and Streptomyces , from Sekem farms in Egypt for their potential to biocontrol RKN and stimulate plant growth in tomato ‘Moneymaker’. The bacteria were compared with well-known microbial biocontrol agents (MBA), including Rhizobium etli G12 (B21), Pseudomonas trivialis 3Re2-7 (B22), Sporosarcina psychrophile Sd4-11 (B23), and B. subtilis Sb1-20 (B24), and a negative control Escherichia coli JM109 (B25). The study involved seed-coated and uncoated plants with bacterial isolates, planted in plastic pots, and inoculated with 1500 M. incognita J 2 individuals per pot. Plants were grown in a saran-house during the 2019 and 2020 fall seasons, and their RKN-satisfying response (number of galls: NG and egg masses: NEM), vegetative growth, and metabolic activity were assessed 45 days after inoculation. Results: In both seasons, seed coating with bacterial isolates achieved a significant improvement in plant growth (coefficient of variation: CV ranging 26.8-120.2% in 2019 and 10.9-48.8% in 2020) and a reduction in RKN-satisfying response (CV for NG: 57.6 and 53.8%, respectively; and for NEM: 56.5 and 65.3%, respectively). Compared to uncoated-seed plants, the bacterial seed coating reduced NG by 0.66-74.09% in 2019 and 14.61-66.29% in 2020. Similarly, NEM decreased by 0.63-70.61% in 2019 and 41.91-77.46% in 2020. The coated-seed plants by Bacillus subtilis subsp. spizizenii (B5), Streptomyces subrutilus Wb2n-11 (B12), Streptomyces scabiei (B19), and Bacillus mojavensis (B20), along with the well-known MBAs B22 and B23, showed increased photosynthetic pigments, fresh weight of roots and shoots, stem size, and number of leaves. This growth has also led to higher dry weights in roots and shoots, and an increase in the root content of carbohydrates and proteins. Seed coating induced systemic RKN resistance by increasing polyphenol in root. In contrast, uncoated-seed plants showed reduced foliar photosynthesis pigment and metabolic activity due to high RKN damage. Principal component analysis revealed significant correlations between the evaluated traits. Hierarchical clustering categorized bacteria isolates into five clusters based on their impact on estimated plant traits. Conclusion: B5, B12, B19, B20, B22, and B23 demonstrated superior performance in both controlling RKN and stimulating vegetative growth in tomato ‘Moneymaker’ plants as known MBAs. Bacillus Meloidogyne Polyphenols Paenibacillus Streptomyces Solanum lycopersicum Figures Figure 1 Figure 2 Figure 3 Figure 4 1. Introduction Tomato ( Solanum lycopersicum L.) is an important vegetable crop worldwide because of its nutritional and processing value, which is essential for food security and nutrition. The production of tomato worldwide in 2023 was estimated 192.32 million tons with an average yield of 35.5 tons hectare − 1 across 5.4 million hectares( https://faostat.fao.org/ ). Egypt was the fifth-largest producer worldwide, yielding 6.28 million tons from 143,618 hectares in 2022, with an average yield of 43.7 tons hectare − 1 ( https://faostat.fao.org/ ). Tomato crop is greatly impacted by plant-parasitic nematodes, which can cause yield losses of 30–40% (Charchar et al. 2003 ), and can even reach 85% in highly susceptible cultivars, particularly in tropical and subtropical regions (Lopes-Caitar et al. 2019 ). Tomato production in Egypt is significantly hindered by the presence of root-knot nematodes (RKN; Meloidogyne spp.), particularly M. incognita (Abd-Algawad 2014 ). Nematode control is a complex and costly process with limited options. Integrated pest management (IPM) for RKN in sustainable agriculture systems involve four main strategies: host plant resistance, agricultural practices, biological control, and chemical control. Chemical nematicides, such as methyl bromide, chloropicrin, 1,3-dichloropropene, or metam-based products (metam sodium and metam potassium), effectively reduce RKN damage, but their cost and potential harmful impacts on human health and the environment often make them less appealing to farmers (Castellano-Hinojosa et al. 2022 ). IPM is now increasingly focused on biological control, which uses antagonistic organisms to reduce nematode populations and their abundance and damage, providing a sustainable alternative to chemical nematicides. Plant-parasitic nematodes can be biocontrolled by using a variety of antagonistic organisms, known as biological control agents (BCAs), including fungi, bacteria, viruses, protists, nematode antagonists, and other invertebrates (Sharma et al. 2025 ). BCAs can directly control RKN through resource competition, predation, and parasitism at various life stages, or indirectly by producing nematicidal metabolites and inducing plant resistance (Sharma et al. 2025 ). The use of BCAs in suppressing nematode populations has been controversial because other soil microorganisms and the host plant can be adversely affected (Sharma et al. 2025 ). Furthermore, beneficial plant–microorganism interactions are commonly exact, with only a few broad-spectrum antagonists identified (Sharma et al. 2025 ). Meloidogyne spp, can be suppressed by indigenous microbial communities in arable soils (Adam et al. 2014 , Amorim et al. 2024 , Sharma et al., 2025 ). Identifying antagonistic microbes and understanding the mechanisms that regulate nematode populations is particularly important in soils that naturally suppress Meloidogyne spp. Rhizosphere bacterial genera such as Bacillus , Paenibacillus , Pseudomonas , and Streptomyces , have been successfully used as BCAs for nematodes in vegetable crops, including tomato (Amorim et al. 2024 , Sharma et al. 2025 ). Rhizosphere bacteria, particularly Bacillus species, have shown the ability to control Meloidogyne spp. in tomato (Hu et al. 2022 , Jiménez-Aguirre et al. 2023 ). Species such as B. megaterium , B. circulans , B. subtilis , B. firmus , and B. pumilus produce enzymes (d’Errico et al. 2019 , Lee et al. 2016, Viaene et al. 2006 ) and bioactive compounds (Ayaz et al. 2021 , Jamal et al. 2017 ) that damage nematodes and inhibit their development, as well as promote plant growth by activating systemic resistance (Adam et al. 2014 , Xia et al. 2019 ) and regulating defense-related genes in tomato plants (Xia et al. 2019 ). The behavior and feeding patterns of nematodes can also be modified by these bacteria (Viaene et al. 2006 ). Strains of B. aryabhattai , B. cereus , B. firmus , B. halotolerans , B. pumilus , B. subtilis , and B. velezensis have been shown to reduce nematode populations, prevent gall formation, and inhibit egg hatching both in vitro and in vivo (Amorim et al. 2024 , Hu et al. 2022 , Khan et al. 2012 , Viljoen et al. 2019 , Xia et al. 2019 , Xiao et al. 2018 ). The reproductive cycle of M. incognita and paralysis in nematode juveniles can be disrupted by B. cereus (Colagiero et al. 2018 , Li et al. 2019 , Xiao et al., 2018 ). Systemic resistance is induced by other strains of B. halotolerans and B. atrophaeus by producing volatile organic compounds (VOCs) that have nematocidal properties (Ayaz et al. 2021 ). Recent research has validated that B. cereus IBCBb 116 significantly reduces nematode populations and enhances tomato plant growth, with the potential for IPM, especially when combined with susceptible tomato genotypes (Amorim et al. 2024 ). Pseudomonas species, particularly P. aeruginosa and P. fluorescens , have shown strong potential in controlling RKN ( M. javanica and M. incognita ) in humid soils (Colagiero et al. 2018 , Hashem and Abo-Elyousr 2011 , Khan et al. 2012 ). P. aeruginosa isolates can inhibit the growth of M. javanica and suppress their populations, as well as prevent root galling by producing hydrogen cyanide (Stirling and Mankau 1978 , Siddiqui et al. 2007 ). P. fluorescens enhanced the defense enzyme activity in RKN-resistant tomato plants and produced antibiotics and enzymes that inhibit nematode egg hatching and reduce egg mass (Khan et al. 2012 ). P. aeruginosa IBCBb 122 showed over 68% mortality in M. incognita juveniles in vitro and effectively reduced nematode populations in greenhouse tomato pots (Amorim et al. 2024 ). Nasima et al. (2002) found that Pseudomonas sp. filtrates can increase juvenile mortality, decrease nematode populations, and improve tomato plant growth and yield. Similarly, Paenibacillus species, like P. polymyxa , P. barcinonensis , and P.alvei , have demonstrated nematicidal effects. P. polymyxa KM2501-1 eliminated M. incognita through contact or fumigation in honey traps (Cheng et al. 2017 ), while P. barcinonensis A10 and P. alvei T30 caused paralysis in M. incognita J 2 juveniles (Viljoen et al. 2019 ). Actinobacteria, especially Streptomyces spp., areabundant in soil and have been demonstaretd to producednematicidal compounds such as actinomycins and chitinase, that directly target nematodes (Jin et al. 2017 , Yoon et al. 2012 , Sharma et al. 2019 ). Species such as S. albogriseolus , S. cacaoi , S. antibioticus , and S. rubrogriseus have been proven through parasitism and toxins production (Jin et al. 2017 , Yoon et al. 2012 ). Actinobacteria also contribute to naturally suppressive soils by anhancing plant growth, suppressing pathogens, and strengthening plant defenses. Antagonistic bacteria that can control nematodes and soil-borne pathogens have been spotted in Egyptian organic farming soils and on the roots of medicinal plants and have been isolated and evaluated for effectiveness. Various bacterial species/isolates showed antagonism against nematodes in vitro studies by Köberl et al. (2010, 2011 , 2013 ) and Adam et al. ( 2014 ). Based on this, this study aimed to evaluate 20 bacterial isolates from the genera Bacillus , Lysobacter , Paenibacillus , and Streptomyces for their potential in biocontrol RKN and promot growth in the susceptible tomato ‘Moneymaker’ under in vivo conditions. Their performance was compared to known effective microbial biological agents: Bacillus subtilis subsp. subtilis Sb1-20, Pseudomonas trivialis 3Re2-7, Rhizobium etli G12, and Sporosarcina psychrophile Sd4-11. 2. MATERIALS AND METHODS 2.1. Nematode culture A pure culture of RKN was established by using naturally infected tomato roots. The Meloidogyne species was determined using the perineal pattern system for mature females as described by Taylon et al. (1956) and Seinhorst (1966). An egg mass from a female of M. incognita was used to inoculate eggplant plants using a perennial pattern. Eggs were extracted from heavily galled roots using a 1.5% chlorine solution in order to prepare nematode inoculum according to Hussey and Barker (1973). Roots were cut, put in a bottle with solution, and shaken strongly for 3 min. The egg suspension was received on the 20 µm sieve, then placed on a modified Baermann dish and incubated at 25±2°C for 7-10 days (Hooper et al. 2005). Hatched J2s were collected daily and stored at 6°C to be used in the experiments. 2.2. Plant materials Susceptible tomato LA2706 ‘Moneymaker’ ( mi-1 / mi-1 ) (https://tgrc.ucdavis.edu/) was used in this study. Tomato 'Moneymaker' plants were cultivated in a greenhouse during the 2019 winter season for seed propagation. 2.3. Bacterial antagonists Twenty-four bacterial isolates of various genera, listed in Table 1, were evaluated for their potential to RKN-suppress and stimulate vegetative growth of tomato ‘Moneymaker’ plants, with Escherichia coli JM109 as the negative control. Bacterial isolates included 20 bacterial isolates (B1-B20) that were previously tested in-vitro for RKN suppression of M. incognita by Köberl et al. (2013), and four bacterial isolates (B21 - B24) have previously shown activity against RKN and soil-borne fungal pathogens (Köberl et al. 2010, 2011, Martinuz et al. 2012, Scherwinski et al. 2008) as positive controls. The bacterial antagonists were cultured on Luria-Bertani (LB) medium in Petri dishes 9 cm in diameter at 27±3 °C and 16/8h photoperiod for 3 days until complete growth on the medium surface. 2.4. Experimental procedure Seeds were thoroughly sterilized in 2% sodium hypochlorite (half strength of commercial bleach) for 10 min on the shaker, washed four times with distilled water, and placed on filter paper to dry under sterile conditions. Fifty seeds were mixed in a bacterial lawn until the bacteria completely coated the seed surface. The uncoated seeds were considered another negative control. The coated seeds were left for a few minutes under a laminar flow hood to dry. The coated and uncoated seeds were sown on September 1 st in the 2019 and 2020 seasons in 209-cell seedling trays filled with a mixture of coconut peat and vermiculite (volume 1:1) enriched with macro and microelements. Seeds were seeded into trays with two empty rows between each coating treatment to prevent cross-contamination of bacterial isolates. The seeds were planted under saran-house conditions at the Department of Agricultural Zoology and Nematology, Faculty of Agriculture, Cairo University (30°00'59.8"N 31°12'21.0"E). Three-week-old seedlings were transplanted into a plastic pot with a 15 cm diameter (seedling/pot), containing 600 g of sterilized a clay: sandy mixture (1:1 volume), and watered to field’s capacity. Pots were arranged in a randomized complete block design (RCBD) with three replicates under saran-house conditions. Each experimental unit contained ten plants. Plants were subjected to common agricultural practices without applying pesticides. After a week, every pot was inoculated with 1,500 freshly hatched J2 in four holes of 2 cm depth at 3 cm distance from the stem base. 2.5. Efficacy of the bacterial isolates on RKN-satisfying response and growth improvement of tomato The traits related to the RKN-satisfying response, vegetative growth, and metabolic activity of ‘Moneymaker’ plants from various seed-coating treatments were assessed 45 days after inoculation (DAI). 2.5.1. The RKN-satisfying response Banora and Almaghrabi (2019) states that the RKN-satisfying response involves the plant’s ability to suppress the development or reproduction of RKN compared to a susceptible plant of the same species (Banora and Almaghrabi 2019). The RKN-satisfying responses of tomato ‘Moneymaker’ plants grown with coated and uncoated seeds were assessed. Nematode populations of each plant’s root system were extracted using the method described by Hooper et al. (2005). The response was evaluated by counting the number of egg masses (NEM) and galls (NG) in a root system. 2.5.2. Plant vegetative growth and metabolic activity Plant measurements were estimated based on the five most representative plants in each EU. Vegetative growth traits included stem length (SL), stem diameter (SD), number of plant leaves (NPL), the area of the fifth fully expanded leaf from the top of the plant (LA), fresh and dry weights of shoots (FSW & DSW) and roots (RFW & RDW), and leaf content of photosynthesis pigments. SL was measured from the soil surface to the stem tip, while SD was measured about 1cm above the soil. The leaf area was measured using the leaf-weighting technique (Pandey and Singh 2011). The photosynthesis pigments were extracted by grinding 0.5g of fresh leaves with 5ml of dimethylformamide according to Moran (1982). Leaf photosynthesis pigments (chlorophyll a: chlor-a, chlorophyll b: chlor-b, total chlorophyll: t-chlor, and total carotenoids: t-carot) were measured using a Jenway 6305 UV/visible spectrophotometer (Jenway, Wales, UK) at 664, 647, and 480 nm (Moran 1982). The formulas for calculating pigment concentrations were as follows: chlor-a = 11.65A 664 – 2.69A 647 , chlor-b = 20.81A 647 – 4.53A 664 , and t-carot = (1000A 480 – 1.42Ca – 46.09Cb)/202. 100g of fresh weight of each shoot and root was oven-dried at 70ºC until a constant weight was reached to measure the dry matter content of shoots (SDM) and roots (RDM). The AOAC method was used to estimate the root content of carbohydrates (Carb), proteins (Pr), and polyphenols (PPh) in dry weight (Latimer 2012). 2.6. Statistical Analysis The Shapiro-Wilk test was performed to check the normality of the data, revealing significant differences in NG, NEM, SL, SFW, RDM, Chlor-a, Chlor-b, Carb, Pr, and PPh in both seasons; NPL, SDW, and RDW in the 2019 season only; and T-chlor and T-Carot in the 2020 season only. Consequently, an arcsine square root transformation was applied to these variables for statistical analysis (Wickens and Keppel 2004). For each season, the variability in the effectiveness of bacterial isolates in suppressing RKN and enhancing vegetative growth of ‘Moneymaker’ tomato plants was assessed using variance components derived from RCBD-ANOVA and the coefficient of variation ( CV = standard deviation / mean). The CV are classified as low (30%) as mentioned by Beah et al. (2021) . A high coefficient of variation indicates significant influence by bacterial isolates. MSTATc v.2.1 software (Michigan State University, Michigan, USA) was used to perform these analyses. Principal component analysis (PCA) and hierarchical cluster analysis (HCA) were used to evaluate variation and similarity/dissimilarity among coating and uncoating seed plants. The data from both seasons were analyzed was combined and analyzed. The data were standardized with Z-scores to account for scale differences before analysis. The latent root criterion (eigenvalue >1) and parallel analysis were used to determine significant components using varimax rotation in PCA (Johnson and Wichern 2007). To assess correlations between traits and the relationships between treatments and traits, the biplot of the top two PCs was constructed according to Yan and Kang (2003). The cosine of the angle between the vectors was used to estimate correlations: an angle of 90° indicates no correlation, 90° indicates a negative correlation. Furthermore, Pearson’s correlation coefficients were used to identify traits that are directly related to RKN-satisfying responses in both coated and uncoated-seed tomato ‘Moneymaker’ plants. HCA was performed using the Euclidean distance and unweighted pair-group average (UPGA) method. Multivariate analyses including Pearson correlation were performed using the IBM SPSS software version 26.0.0 (SPSS Inc., Chicago, Illinois, USA) and XLSTAT software version 2019 (Addinsoft, Paris, France). 3. RESULTS AND DISCUSSION 3.1. RKN-satisfying response In both seasons, seed ‘Moneymaker’ coating with several bacterial antagonists had a highly significant effect ( P <0.001) on the RKN-satisfying responses, i.e., NG (966.826 *** and 342.406 *** , respectively) and NEM (921.924 *** and 488.081 *** , respectively), of their plants in both seasons (Figure 1). This was further supported by the notably very high CV values for both NG (57.6 and 53.8%, respectively) and NEM (56.5 and 65.3%, respectively) in each season (Figure 1). NG on the roots of ‘Moneymaker’ plants ranged from 15.68 with B23-coated seed plants to 60.51 with uncoated seed plants in 2019, and from 15.00 with B23-coated seed plants to 44.50 with uncoated seed plants in 2020 (Figure 1). The highest NG in both seasons was found with uncoated seed plants (60.51 & 44.50, respectively) and B11-coated seed plants (60.11 & 38.00, respectively), with significant similarity ( P <0.05) among them in 2019 only. The lowest NG in both seasons were with coated seed plants in B23 (15.68 & 15.00, respectively) and B19 (21.27 & 16.00, respectively), with significant similarities ( P <0.05) among them in 2020 only. NEM per root of ‘Moneymaker’ plants ranged from 18.26 with B23-coated seed plants to 61.74 with B11-coated seed plants in 2019 and from 15.33 with B23-coated seed plants to 68.00 with uncoated plants in 2020 (Figure 1). The uncoated seed plants had the highest NEM during both seasons (62.13 & 68.00, respectively) with significant similarities ( P <0.05) with coated seed plants by B5, B11, and B25 in 2019 only (61.22, 61.74, and 61.03, respectively). B23-coated seed plants showed the lowest NEM in both seasons (18.26 & 15.33, respectively), significantly similar to B19-coated seed plants in 2020 (16.50). The B22-coated plants were ranked in the next significantly lowest NEM (23.51 & 18.00, respectively), followed by coated seed plants by B13 (26.76&19.75, respectively) and B24 (26.56 & 19.00, respectively). Seed coating with bacterial antagonists enhanced the resistance of ‘Moneymaker’ plants to RKN, showing a reduced RKN-satisfying response in coated seed plants compared to uncoated ones (Figure 1). The presence of bacterial strains of Bacillus , Paenibacillus , and Pseudomonas in tomato can result in systematic resistance to Meloidogyne spp. (Siddiqui et al. 2007, Adam et al. 2014, Xia et al. 2019, Ghahremani et al. 2020). B. subtilis Sb4-23, Mc5-Re2, and Mc2-Re2 (Adam et al. 2014), and B. velezensis YS-AT-DS1 (Hu et al. 2022) can reduced M. incognita J2 infection and related damage, including NG and NEMs. The inhibition of gall formation, egg hatching, and J2 activity of M. incognita was found to be mediated by B. cereus Jdm1, according to Xiao et al. (2018) findings. Colagiero et al. (2018) showed that B. cereus , B. licheniformis , Paenibacillus fluoresencs, and P. brassicacearum disrupted the reproductive cycle of M. incognita . The presence of B. cereus BCM2 prevented M. incognita juveniles from reaching tomato plants, as reported by Li et al. (2019). According to Viljoen et al. (2019), B. firmus T11, B. aryabhattai A08, Paenibacillus barcinonensis A10, P. alvei T30, and B. cereus have paralyzed M. incognita J2 . According to Ghahremani et al. (2020), B. firmus I-1582 was responsible for killing Meloidogyne spp. nematodes and prevented their egg from hatching. Ayaz et al. (2021) found that B. atrophaeus GBSC56 produced VOCs, which induced severe oxidative stress in M. incognita and enhanced the activity of antioxidant enzymes in infested tomato roots. 3.2. Vegetative growth performance RKN infection in tomato plants causes galls to form in the root system, disrupting nutrient and water uptake. These galls also create feeding sites in the vascular tissues, which harbor adult female nematodes. This results in symptoms of yellowing leaves, wilting plants and stunting their growth, floral abortions, reduced fruit quantity and quality, and, in severe cases, plant death (Banora 2023). Evaluating vegetative growth traits is an important indicator of plant resistance to RKN. The vegetative growth performance of tomato ‘Moneymaker’ coated and uncoated seed plants at 45 DAI under RKN infestation is presented in Table 2. Seed ‘Moneymaker’ coating with several bacterial antagonists had a highly significant effect ( P <0.001) on the plant’s vegetative growth performance under RKN infestation conditions in both seasons (Table 2). This was further confirmed by the relatively moderate to very high CV observed for vegetative traits in each season, ranging 16.45-60.37% in 2019 and 10.94-51.28% in 2020 (Table 2). SL (cm) of ‘Moneymaker’ plants ranged from 30.67 with uncoated seed to 109.03 with coated seed plants by B16 and B17 in 2019, and from 50.00 with uncoated seed plants to 74.72 with coated seed plants by B6 and B16 in 2020. The uncoated seed plants had the shortest stems in both seasons (30.67 & 50.00, respectively), with significant similarities with coated seed plants by B1, B8, B10, B11, B18, and B22 in 2019 (ranging from 30.70 - 53.68). B16-coated seed plants had the tallest plants in both seasons (109.03 & 74.72, respectively), with significant similarities ( P <0.05) with coated plants by B4, B14, B15, B17, B21, and B24 in 2019 (ranging from 82.92 – 109.03) and those by B6, B9, and B20 in 2020 (74.72, 73.13, and 73.17 cm, respectively). SD (mm) of ‘Moneymaker’ plants ranged from 2.40 with uncoated plants to 5.13 with B19-coated seed plants in 2019 and from 4.52 with B9-coated seed plants to 5.80 with B19-coated seed plants in 2020 (Table 2). The uncoated seed plants have the thinnest stems in both seasons (2.40 & 4.70, respectively), with significant similarities ( P <0.05) with coated seed plants by B2, B9, B15, and B16 in 2020 only (4.63, 4.52, 4.62, and 4.75, respectively). B19-coated seed plants had the thickest stems in both seasons (5.13 & 5.80, respectively), with significant similarity ( P <0.05) with those coated by B9 in 2019 only (5.07), and B11 and B14 in 2020 (5.57 & 5.65, respectively). NPL of ‘Moneymaker’ plants ranged from 5.0 with uncoated seed plants and B1-coated seed plants to 12.4 with B14-coated plants in 2019, and from 7.17 with B12-coated seed plants to 11 with B22-coated seed plants in 2020 (Table 2). The lowest NPL in both seasons was found with uncoated seed plants (5.0 and 8.0, respectively) and B1-coated seed plants (5.0 & 8.0, respectively), with significant similarities ( P <0.05) among them and coated seed plants by B2, B9, B11, B15, B16, B19, and B25 in 2020 (ranging from 8.17 – 8.67). B4-coated seed plants had the most leaves in both seasons (12.40 & 10.83, respectively), with significant similarities ( P <0.05) with coated seed plants by B4, B14, and B17 in 2019 (12.0, 12.40, and 12.0, respectively), and those by B22 in 2020 (11.0). SFW (g) of ‘Moneymaker’ plants ranged from 28.13 with uncoated seed plants to 55.98 with B23-coated seed plants in 2019, and from 34.08 with uncoated seed plants to 61.00 with B23-coated seed plants (Table 2). The uncoated seed plants had the lowest SFW in both seasons (28.13 & 34.08, respectively), followed by coated seed plants by B12 (29.92 & 35.93, respectively), B15 (29.57 & 35.67, respectively), and B22 (29.40 & 35.50, respectively), with significant similarities ( P <0.05) among them in only 2020, and coated seed plants by B1, B3, B7, B8, B13, B14, B21, and B25 in 2020 (ranging from 36.83 – 38.12). B23-coated seed plants had the highest SFW in both seasons (55.98 & 61.00, respectively), followed by B17-coated seed plants (49.20&55.55, respectively), and then B18-coated seed plants (45.55 & 51.82, respectively). SDW (g) of ‘Moneymaker’ plants ranged from 3.49 with uncoated seed plants to 7.06 with B23-coated seed plants in 2019, and from 4.01 with uncoated seed plants to 8.21 with B23-coated seed plants (Table 2). The uncoated seed plants had the lowest SDW in both seasons (3.49&4.01, respectively), followed by coated seed plants by B1 (3.65 & 4.02, respectively) and B22 (3.74 & 4.06, respectively), with significant similarities ( P <0.05) among them in 2020, and coated seed plants by B7 and B25 in 2020 (4.00 & 4.61, respectively). In both seasons, B23-coated seed plants had the highest SDW (7.06 & 8.21, respectively), followed by B17-coated seed plants (6.21 & 6.92, respectively). RFW (g) ranged from 1.80 with uncoated seed plants to 9.23 with B20-coated seed plants in 2019, and from 3.03 with uncoated seed plants to 10.46 with B20-coated seed plants in 2020 (Table 2). The uncoated seed plants had the lowest RFW in both seasons (1.80 & 3.03, respectively), with significant similarities ( P <0.05) with coated seed plants by B1, B3, B13, B21, and B25 in only 2020 (4.25, 4.27, 3.59, 4.59, and 4.43, respectively). In both seasons, B20-coated seed plants had the highest RFW (9.23&10.46, respectively), followed by coated seed plants by B12 (7.18 & 8.41, respectively), and then B19 (6.93 & 8.16, respectively). RDW (g) ranged from 0.40 with uncoated seed plants to 1.97 with B20-coated seed plants in 2019, and from 0.68 with uncoated seed plants to 2.31 with B20-coated seed plants in 2020 (Table 2). The lowest RDW in both seasons was found with uncoated seed plants (0.40 & 0.68, respectively) and B13-coated seed plants (0.42 & 0.70 g, respectively), with significant similarities ( P <0.05) among them and coated seed plants by B1, B3, B7, and B25 in 2020 (0.91, 0.92, 0.95, and 0.97, respectively). In both seasons, B20-coated seed plants had the highest RDW (1.97 & 2.31, respectively), followed by coated seed plants by B12 (1.52 & 1.56, respectively) and B15 (1.49 & 1.64, respectively), then coated seed plants by B10 (1.41 & 1.79, respectively), B11 (1.43 & 1.56, respectively), and B18 (1.44 & 1.79, respectively). 3.3. Foliar photosynthesis pigments The foliar photosynthesis pigment content of tomato ‘Moneymaker’ coated and uncoated seed plants is presented in Table 3. Seed ‘Moneymaker’ coating with several bacterial antagonists had a highly significant effect ( P <0.001) on the leaf content of photosynthesis pigments under RKN infestation conditions in both seasons (Table 3). Further support was provided by the consistently high to very high CV values recorded for the leaf content of photosynthesis pigments across seasons, which ranged from 27.07-120.19% in 2019 and from 23.49-32.57% in 2020 (Table 3). The leaf chlor-a content (mg g -1 ) ranged from 0.58 with B11-coated seed plants to 2.13 with B14-coated seed plants in 2019, and from 0.78 with B13-coated seed plants to 1.54 with B21-coated seed plants in 2020 (Table 3). Leaf chlor-a content was inconsistent during both seasons. The highest leaf chlor-a content was found in B14-coated seed plants in 2019 (2.13), while in coated seed plants by B21 and B14 in 2020 (1.54 & 1.53, respectively) with significant similarities ( P <0.05) among them. In 209, the lowest leaf chlor-a content was found in 2019 with coated seed plants by B1, B2, B10, B11, and B12 (ranging from 0.58 - 0.79), with significant similarities ( P <0.05) among them. In 2020, uncoated seed plants and coated seed plants by B4, B13, B17, B18, and B24 had the lowest leaf chlor-a content (ranging from 0.78 - 0.89). Leaf chlor-b content (mg g -1 ) ranged from 0.05 with uncoated seed plants to 0.81 with B23-coated seed plants in 2019, and from 0.26 with B6-coated seed plants to 0.46 with B23-coated seed plants in 2020 (Table 3). B23-coated seed plants had the highest leaf chlor-b content in both seasons (0.81 & 0.46, respectively), with significant similarity ( P <0.05) with B21-coated seed plants in 2020 (Table 3). Uncoated seed plants had the lowest leaf chlor-b content in both seasons (0.05 & 0.28, respectively), with significant similarities ( P <0.05) with seed-coated seed plants by B10 and B20 in 2019 (0.07 & 0.10, respectively), and coated seed plants by B4, B6, B9, B13, B17, B18, and B24 in 2020 (ranging from 0.26 – 0.28). Leaf total t-chlor content (mg g -1 ) was between 0.73 with B10-coated seed plants and 2.26 with B14-coated seed plants in 2019, and between 1.06 with B13-coated seed plants and 1.95 with B21-coated seed plants in 2020 (Table 3). Leaf t-chlor content was inconsistent in both seasons, particularly the highest content. The highest content was found in B14-coated seed plants in 2019 (2.26), and in coated seed plants by B21 and B23 (1.95 & 1.94, respectively) in 2020. The uncoated seed plants had the lowest t-chlor content in both seasons (0.76 & 1.10, respectively), with significant similarities ( P <0.05) with coated seed plants by B2, B10, and B11 in 2019 (0.83, 0.73, and 0.76, respectively), and coated seed plant by B4, B17, B18, B13, and B24 in 2020 (ranging from 1.06 - 1.16). Leaf t-car (mg g -1 ) was between 0.40 with uncoated seed plants to 0.75 with coated seed plants by B1 and B17 in 2019, and between 0.65 with B24-coated seed plants to 1.05 with B11-coated seed plants in 2020 (Table 3). B11-coated seed plants had the highest leaf t-car content in both seasons (0.71 & 1.05, respectively), with significant similarities ( P <0.05) with coated seed plants by B1 and B17 in 2019 (0.75 for both), and coated seed plants by B21 in 2020 (1.04) (Table 3). The uncoated seed plants had the lowest leaf t-car content in both seasons (0.40 & 0.67, respectively), with significant similarities ( P <0.05) with B21-coated seed plants in 2019 (0.43), and with coated seed plants by B23 and B24 in 2020 (0.67 & 0.65, respectively). Seed-coating tomato ‘Moneymaker’ plants with some bacterial antagonists improved vegetative growth under RKN infestation. The vegetative growth performance of uncoated seed plants was the lowest, while coated seed plants showed enhanced vegetative growth, with some bacterial isolates yielding better results. Coated seed plants by B5, B12, B19, B20, B22, and B23 exhibited favorable vegetative growth traits. The RKN-satisfying response of tomato ‘Moneymaker’ plants was diminished by seed coating with bacterial antagonists (Figure 1), resulting in improved vegetative growth. This was achieved by improving the root's ability to absorb water and nutrients under RKN stress (Farahat et al. 2012). As a result, the coated seed plants showed increased fresh weights of roots and shoots, along with greater stem length and diameter (Table 2). Additionally, the number of plant leaves (Table 2) and their photosynthetic pigment content (Table 3) increased, resulting in a larger green surface area for photosynthesis. Therefore, the fresh and dry weights of the plant shoots and roots increased (Table 2). Furthermore, some bacterial isolates may also have a stimulating effect on vegetative growth genes. Ayaz et al. (2021) found that the genesSlCKX1, SlIAA1, and Exp18 that promote plant growth were found to increase in expression in tomato plants treated with B. atrophaeus GBSC56. 3.4. Plant metabolism activity The plant metabolic activity, i.e., root and shoot content of dry matter and root content of carbohydrates, proteins, and polyphenols, of tomato ‘Moneymaker’ coated and uncoated seed plants at 45 DAI presented in Table 4. The seed coating with several bacterial antagonists had a highly significant effect ( P <0.001) on the metabolic activity of the ‘Moneymaker’ plant under RKN infestation conditions in both seasons (Table 4). However, the CV values for the plant metabolic activity estimates were generally low, except for the root content of Pr in both seasons (23.29 and 14.98%, respectively), as well as SDM and RDM in 2019, which showed very high values (45.73 and 44.06%, respectively; Table 4). SDM (mg g -1 ) ranged from 12.17 with coated seed plants by B1 and B2 to 13.04 with B14-coated seed plants in 2019, and from 10.49 with B7-coated seed plants to 15.98 with B18-coated seed plants in 2020 (Table 4). In both seasons, the lowest SDM was observed with coated seed plants by B1 (12.17 & 10.90, respectively) and B7 (12.52 & 10.49, respectively), with significant similarities ( P <0.05) among them and uncoated seed plants and coated seed plants by B2, B21, B9, and B10 in 2019 (12.38, 12.17, 12.45, 12.39, and 12.45, respectively), and those by B22 in 2020 (11.46). The highest SDM in both seasons was found with coated seed plants by B5 (12.72 & 15.44, respectively) and B19 (13.03 & 15.98, respectively), with significant similarities ( P <0.05) among them and coated seed plants by B1, B4, B6, B8, B12, B14, B16, B17, B22, B24, and B25 in 2020 only (ranging from 12.64 – 13.04). RDM (mg g -1 ) ranged from 21.03 with B14-coated seed plants to 22.60 with B7-coated seed plants in 2019, and from 18.57 with B4-coated seed plants to 30.94 with B18-coated seed plants in 2020. The highest RDM in both seasons was found with B8-coated seed plants (22.54 & 26.40, respectively), with significant similarities ( P <0.05) with uncoated seed plants and coated seed plants by B2, B7, and B21 in 2019 (22.20, 21.98, 22.60, and 22.43, respectively), and those by B9 and B19 in 2020 (29.03 & 30.94, respectively). In both seasons, the lowest RDM was found with coated seed plants B1, B3-B6, B10–B13, B14-B16, B18, B22, B23, B24, and B25 with significant similarities ( P <0.05) among them and coated plants by B2, B9, B17, and B19 in 2019 (21.98, 21.56, 21.54, and 21.54, respectively), and uncoated and coated seed plants by B21 and B7 in 2020 (22.43, 23.53, and 18.70, respectively) (Table 4). Results of the root content of carbohydrates, proteins, and polyphenols in tomato ‘Moneymaker’ coated and uncoated seed plants are presented in Table 4. Coating the seed ‘Moneymaker’ with several bacterial antagonists had a highly significant effect ( P <0.001) on the root content of Carb, Pr, and PPh under RKN infestation conditions in both seasons (Table 4). The root Carb content (mg g -1 ) was between 41.56 with uncoated seed plants and 53.97 with B21-coated seed plants in 2019, and between 38.81 with uncoated seed plants and 50.31 with B19-coated seed plants in 2020 (Table 4). The root Carb content was inconsistent in both seasons, particularly with the highest content. In 2019, the highest root Carb content was found in B21-coated seed plants (53.97), followed by coated seed plants by B6 (53.59) and B13 (53.58), which have significant similarities ( P <0.05) among them, and then coated seed plants by B5, B9, and B19 (53.28, 53.25, and 53.25, respectively), which have significant similarities ( P <0.05) among them (Table 4). B19-coated seed plants had the highest root Carb content in 2020 (50.31), followed by B4-coated seed plants (50.17), and then B20-coated seed plants (50.05). The uncoated seed plants had the lowest root Carb content in both seasons (41.56 & 38.81, respectively), with significant similarities with coated seed plants by B1, B2, B8, and B10 in 2019 only (51.57, 51.73, 51.76, and 51.08, respectively), and coated seed plants by B5, B15, B18, and B23 in 2020 (48.40, 48.37, 47.85, and 48.29, respectively). The root Pr content (mg g -1 ) ranged from 8.81 with B5-coated seed plants to 19.10 with B18-coated seed plants in 2019, and from 10.07 with uncoated seed plants to 17.82 with B18-coated seed plants (Table 4). The highest root Pr content was found in B18-coated seed plants in both seasons (19.10 & 17.82, respectively), with significant similarities with coated seed plants by B10 and B15 in 2019 (19.09 & 19.08, respectively), and with B11-coated seed plants in 2020 (17.81). The lowest root Pr content was inconsistent in both seasons. In 2019, B5-coated seed plants and uncoated seed plants had the lowest content (8.81 and 10.80, respectively), with significant differences ( P <0.05) among them. In 2020, the uncoated seed plants had the lowest content (10.07), followed by B19-coated plants (15.69). The root PPh content (mg g -1 ) ranged from 5.16 with uncoated seed plants to 6.74 with B21-coated seed plants in 2019, and from 4.82 with uncoated seed plants to 6.30 with B2-coated seed plants in 2020 (Table 4). The root PPh content was inconsistent in both seasons, particularly the highest content. The highest root PPh content was found in coated seed plants by B21 and B13 in 2019 (6.74 & 6.73, respectively), with significant similarities ( P <0.05) among them, and coated seed plants by B2, B17, and B19 in 2020 (6.30, 6.30, and 6.29, respectively) with significant similarities ( P <0.05) among them. The uncoated seed plants had the lowest root PPh content in both seasons (5.16 & 4.82, respectively). RKN infestation in uncoated seed tomato ‘Moneymaker’ plants resulted in a decrease in foliar photosynthetic pigments (Table 3), causing a decrease in photosynthesis and the production of carbohydrates and proteins (Khan et al. 2006, Campos et al. 2012). RKN also increased reactive oxygen species (ROS; Teresa Melillo et al. 2006), which elevated chlorophyllase activity, causing chlorophyll degradation and impairing cellular structures such as proteins, carbohydrates, lipids, and DNA (Fujimoto et al. 2021, Holbein et al. 2016). Therefore, leaves of uncoated seed ‘Moneymaker’ plants have a higher chlor-a/chlor-b ratio in the leaves, likely due to chlorophyll-b being degraded to chlorophyll-a (Fang et al. 1998). RKN infestation also reduced dry matter content in both roots and shoots and lowered the carbohydrate and protein content in the roots, consistent with previous studies (Khan et al. 2022, Banora and Almaghrabi 2019). RKN-induced secretion of hydrolyzing enzymes may contribute to the decreased carbohydrate content in the roots (Anwar 1995, Farahat et al. 2012), while increased root protein content may be related to resistance mechanisms in the plants (Mukherjee et al. 2020, Patel and Patel 1995). Polyphenols in resistant tomato plants are essential in defending against RKN by acting as signaling molecules to detect and neutralize ROS (Khan et al. 2022, Patel et al. 2017, Campos et al. 2012, Farahat et al. 2012) and by processing nematicidal properties that reduce RKN damage (Pratysha 2022, Aissani et al. 2018). In this study, coated-seed tomato ‘Moneymaker’ plants exhibited higher polyphenol contents in their roots compared to those with uncoated seeds. As a result, the coated-seed plants showed lower NG and NEM, as well as a higher content of foliar photosynthetic pigments, dry matter content in both shoots and roots, and root carbohydrates and proteins. Their ability to scavenge ROS may be the reason for this. 3.8. Multivariate analyses Multivariate analyses, particularly principal component analysis (PCA) and cluster analysis (CA), are often employed to evaluate variability among treatments, identify the correlations between traits and treatments, and categorize treatments based on estimated traits to select the best ones (Johnson and Wichern 2007). PCA is a valuable tool for dividing multiple traits into fewer components that explain the majority of the variance. In this study, PCA was performed on 18 traits related to RKN-satisfying response, vegetative growth, and metabolic activity of uncoated and coated seed ‘Moneymaker’ plants by several bacterial antagonists grown in a naturally RKN-infested greenhouse during the 2019 and 2020 fall seasons. Eighteen principal components (PCs) were identified with eigenvalues ranging from 5.65 to 0.00009, as shown in Figure 2. The first 6 PCs had eigenvalues >1 and contributed 94.08% of cumulative variability (Figure 2). All estimated traits had a significant impact on the first 6 PCs (Table 5), except PC5, suggesting that these traits of ‘Moneymaker’ plants can be used to classify bacterial isolates (Johnson and Wichern 2007). PC1 (eigenvalue= 5.472) explained 30.40% of the variance (Figure 2) and was influenced negatively by traits SL (-0.225), SD (-0.240), NPL (-0.258), SFW (-0.225), SDW (-0.268), Chlor-b (-0.207), Carb (-0.331), Pr (-0.276), and PPh (-0.280), and being positively influenced by NG (0.310) and NEM (0.343) (Table 5). PC2 (eigenvalue = 2.395) explained 13.31% of the variance (Figure 2) and was positively correlated only with T-Carot (0.495) (Table 5). PC3 (eigenvalue = 2.228) was responsible for 12.74% of the variance (Figure 2) and had positive correlations with Chlor-a (0.460) and T-Chlor (0.505), but negative correlations with only RFW (-0.374) (Table 5). PC4 (eigenvalue = 1.934) was responsible for 10.743% of the variance (Figure 2) and had positive correlations with RDW (0.516) and RDM (0.435) (Table 5). PC5 (eigenvalue = 1.496) accounted for 8.31% of the variance (Figure 2) but was not influenced by any traits (Table 5). PC6 (eigenvalue = 1.098) explained 6.098% of the variance (Figure 2) and had a positive correlation with only SDM (0.508) (Table 5). Figure 3 displays the biplot of the first two PCs, which depicts the distribution of coated and uncoated seed ‘Moneymaker’ plants with bacterial isolates and the estimated traits under RKN infestation during the 2019 and 2020 fall seasons. The vectors for NG and NEM were grouped in a separate quadrant with similar directions, and angles between them less than 90°. The vectors for the residual traits were located in two quadrants that were opposite to each other, with the vectors close together and angled similarly (Figure 3). Thus, Pearson’s correlation coefficients were estimated. Significant correlations were discovered in Pearson’s correlation coefficients for estimated traits, as shown in Table 6. This study focused on the correlations between RKN-satisfying response, i.e., NG & NEM, and traits that related to vegetative growth and plant metabolic activity in coated and uncoated seed plants. Estimates of NG & NEM showed highly positive correlations among them. Also, NG & NEM showed negative correlations with SL (-0.429 * &-0.511 * , respectively), SFW (0.430 * & 0.408 * , respectively), SDW (0.437 * & 0.421 * , respectively), chlor-b (0.484 * & 0.486 * , respectively), Carb (0.395 * & 0.501 * , respectively), and Pr (0.440 * & 0.529 * , respectively) (Table 6). The photosynthetic pigment content in tomato leaves is reduced by RKN infestation, decreasing the photosynthesis rate and the accumulation of carbohydrates and proteins (Campos et al. 2012, Khan et al. 2006, 2022). Tomato plants are weakened by this, resulting in smaller leaves, shorter stems, and decreased yield components (Banora and Almaghrabi 2019). Previous studies have found negative correlations between tomato plant growth parameters and initial RKN population density (Mekete et al. 2003, Schomaker et al. 2006) and RKN reproduction factor (Gharabadiyan et al. 2013). Padilla-Hurtado et al. (2022) found a significantly negative correlation between the RKN-damage scale and the number of plant fruits. Sharma et al. (1990) and Patel et al. (2017) indicated that polyphenol synthesis in tomato plants is correlated with nematode infection. The coated and uncoated seed plants with several bacterial isolates were distributed across all quarters of the biplot (Figure 3), reflecting high variability among the bacterial isolates impact on RKN-satisfying response, vegetative growth, and metabolic activity of tomato ‘Moneymaker’Top of Form . Treatments with higher trait values were positioned distant from the vector line, often at the vertices of the convenx hull (Johnson and Wichern 2007). Uncoated seed plants were plotted in a separate quadrant. The coated seed plants by B1, B2, B3, B5, B7, B8, B9, B10, B11, B22, and B25 were plotted in another quadrant. The direction of both groups was toward NG and NEM vectors. Coated seed plants by B6, B12, B21, B15, and B20 were plotted in the same quadrant close to the vector lines of SD, SL, T-Carot, SDM, RFW, RDW, RDM, Carb, PPh, and Pr. Coated seed plants by B4, B13, B14, B16, B17, B18, B19, and B24 were plotted in another quadrant close to the vector lines of NPL, Chlor-a, Chlor-b, T-Chlor, SFW, and SDW. Therefore, hierarchical cluster analysis (HCA) can be performed to categorize bacterial isolates based on their impacts on RKN-satisfying response, vegetative growth, and metabolic activity of tomato ‘Moneymaker’ plants (Johnson and Wichern 2007). HCA revealed five distinct clusters as shown in Figure 4. Cluster-1 included only coated seed plants by B25 (Figure 4), and was characterized by very low values of SL and NPL; low values of SFW, SDW, SDM, FRW, RDW, and RDM; moderate values of Chlor-a, Chlor-b, T-Chlor, and SDM; high NG and NEM; and very high values of SD, T-Carot, Carb, Pr, and PPh (Table 7). Cluster-2 consisted of coated seed plants by B1, B3, B5, B6, B7, B8, B9, B10, B11, B12, and B21 (Figure 4), which had low values of Chlor-b and SD, moderate values of NG, NEM, SL, SFW, SDW, RFW, RDW, Carb, Pr, and PPh; and high values of NPL, Chlor-a, T-Chlor, T-Carot, and SDM (Table 7). Cluster 3 consisted of coated seed plants by B2, B4, B13, B14, B15, B16, B17, B18, B19, B20, B22, and B24 (Figure 4), and was characterized by low values of NG and NEM; moderate values of SD, NPL, Chlor-a, Chlor-b, T-Chlor, T-Carot, and RDM; high values of SL, SFW, SDW, SDM, Carb, Pr, and PPh; and very high values of RFW and RDW (Table 7). Cluster 4 consisted of only coated seed plants by B23 (Figure 4), which had very low values of NG, NEM, and RDM; low values of T-Carot, Carb, and PPh; high values of SD, RFW, RDW, and Pr; and very high values of SL, NPL, Chlor-a, Chlor-b, T-chlor, SFW, SDW, and SDM (Table 7). Cluster 5 consisted of only uncoated plants (Figure 4), and was characterized by very high values of NG, NEM, and RDM, and vary low values of the rest traits (Table 7). Cluster centroids had different levels of separation, with the smallest between Clusters 2 and 3 (23.862) and the largest between Clusters 4 and 5 (82.553) (Table 7). The difference in trait profiles between clusters was emphasized by the variance, which ranged from 0 for Clusters 1, 4, and 5 to 198.116 for Cluster-3 (Table 7). Conclusion The bacterial isolates Bacillus subtilis subsp. spizizenii Sd1-14, Streptomyces subrutilus Wb2n-11, Streptomyces scabiei WB1n-4, and Bacillus mojavensis Sd2Re-10 demonstrated strong effectiveness in both suppressing RKN and enhancing vegetative growth in tomato ‘Moneymaker’ plants, as did well-known microbial biocontrol agents Pseudomonas trivialis 3Re2-7 and Sporosarcina psychrophila Sd4-11. Future research will evaluate the effects of these bacterial isolates, individually or in combination, on the growth of various tomato cultivars and other vegetable crops. The focus of the research will be on their potential for manage nematodes and soil-borne pathogens, while also promoting plant growth and maintaining soil microbal communities. Future studies will also aim to assess the extent and duration of soil colonization by these bacterial isolates to identify stable commercial formulations of them in line with national regulations. Declarations Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Funding: Not applicable. Acknowledgments: Not applicable. Author Contribution AMAM is responsible for conceptualization, formal analysis, software, supervision, validation, visualization, and writing of an original draft, review, and editing. AASAE is responsible for the conceptualization and supervision. MA is responsible for the conceptualization and supervision. MAAM is responsible for data curation, investigation, methodology, and writing an original draft. References Abd-Algawad MMM (2014) Plant-parasitic nematode threats to global food security. J Nemat 46:130. Adam M, Westphal A, Hallmann J, Heuer H (2014) Specific microbial attachment to root-knot nematodes in suppressive soil. Appl Environm Microbiol 80:2679-2686. Aissani N, Balti R, Sebai H (2018) Potent nematicidal activity of phenolic derivatives on Meloidogyne incognita . J Helmin 92:668-673. https://doi.org.10.1017/S0022149X17000918. Ali NI, Siddiqui IA, Shaukat SS, Zaki MJ (2002) Nematicidal activity of some strains of Pseudomonas spp. Soil Biol Biochem 34:1051-1058. https://doi.org/10.1016/S0038-0717(02)00029-9. Amorim DJ, Tsujimoto TF, Baldo FB, Leite LG, Harakava R, Wilcken SRS, Gabia AA, Amorim DJ (2024) Bacillus , Pseudomonas and Serratia control Meloidogyne incognita (Rhabditida: Meloidogynidae ) and promote the growth of tomato plants. Rhizoshpere 31:100935. https://doi.org/10.1016/j.rhisph.2024.100935 Anwar SA (1995) Influence of Meloidogyne incognita , Paratrichodours minor , and Pratylenchus scribneri on root-shoot growth and carbohydrates partitioning in tomato. Pakistan J Zool 27:105-113. Ayaz M, Zhao JT, Zhao W, Chi YK, Ali Q, Ali F, Khan AR, Yu Q, Yu JWW, Wu WC, Qi RD, Huang WK (2021) Biocontrol of plant parasitic nematodes by bacteria and fungi: A multi-omics approach for the exploration of novel nematicides in sustainable agriculture. Front Microbiol 15:1433716. https://doi.org/10.3389/fmicb.2024.1433716. Banora MY, Almaghrabi OAA (2019) Differential response of some nematode-resistant and susceptible tomato genotypes to Meloidogyne javanica infection. J Plant Protec Res 59:113-123. https://doi.org/10.24425/jppr.2019.126040. Banora MY (2023) I,pacting of root-knot nematodes on tomato: Current status and potential horizons for its managing. In: Lops F (ed), Tomato Cultivation and Consumption – Innovation and Sustainability. IntechOpen Limited, London, UK, pp.1-22. https://doi.org/10.5772/intechopen.112868. Beah A, Kamara AY, Jibrin JM, Akinseye FM, Tofa AI, Adam AM (2021) Simulating the response of drought-tolerant maize varieties to nitrogen application in contrasting environments in the Nigeria Savannas using the APSIM model. Agronomy 11(1):76. https://doi.org/10.3390/agronomy11010076. Campos VAC, Machado ART, Oliveira DF, Campos VP, Chages RCR, Nunes AS (2012) Change in metabolites in plant roots after inoculation with Meloidogyne incognita . Nematology 14:579-588. https://doi.org/10.1163/15684111x614494. Castellano-Hinojosa A, Noling JW, Bui HX, Desaeger JA, Strauss SL (2022) Effect of fumigants and non-fumigants on nematode and weed control, crop yield, and soil microbial diversity and predicted functionality in a strawberry production system. Science of the Total Environment 852:158285. https://doi.org/10.1016/j.scitotenv.2022.158285. Charchar JM, Gonzaga V, Giordano LB, Boiteux LS, dos Reis NV, de Aragão, FA (2003) Reaction of tomato cultivars to infection by a mixed population of M. incognita race 1 and M. javanica in the field. Nemat Bras 27:49-54. Cheng W, Yang J, Nie Q, Huang D, Yu C, Zheng L, Cai M, Thomashow L, Weller DM, Yu Z, Zhang J (2017) Volatile organic compounds from Paenibacillus polymyxa KM2501-1 control Meloidogyne incognita by multiple strategies. Sci Rep 24:16213. https://doi.org/10.1038/s41598-017-16631-8. Colagiero M, Rosso LC, Ciancio A (2018) Diversity and biocontrol potential of bacterial consortia associated to root-knot nematodes. Biol. Control 120:11–16. https://doi.org/10.1016/j.biocontrol.2017.07.010. d’Errico G, Marra R, Crescenzi A, Davino SW, Fanigliulo A, Woo SL, Lorito M 2019) Integrated management strategies of Meloidogyne incognita and Pseudopyrenochaeta lycopersici on tomato using a Bacillus firmus -based product and two synthetic nematicides in two consecutive crop cycles in greenhouse. Crop Protect 122:159–164. https://doi.org/10.1016/j.cropro.2019.05.004. Fang Z, Bouwkamp J, Solomos T (1998) Chlorophyllase activities and chlorophyll degradation during leaf senescence in non-yellowing mutant and wild type of Phaseolus vulgaris L. J Exp Bot 49:503–10. https://doi.org/10.1093/jxb/49.320.503. Farahat AA, Alsayed AA, El-Beltagi HS, Mahfoud NM (2012) Impact of organic and inorganic fertilizers on nematode reproduction and biochemical alternations on tomato. Not Sci Biol 4:48-55. Fujimoto T, Abe H, Mizukubo T, Seo S. 2021. Phytol, a constituent of chlorophyll, induces root-knot nematode resistance in Arabidopsis via the ethylene signaling pathway. Mol Plant Mic Inter 34:279-285. https://doi.org/10.1094/MPMI-07-20-0186-R. Ghahremani Z, Escudero N, Beltrán-Anadón D, Saus E, Cunquero M, Andilla J, Loza-Alvarez P, Gabaldón T, Sorribas FJ (2020) Bacillus firmus strain I-1582, a nematode antagonist by itself and through the plant.Front Plant Sci 11:796. https://doi.org/10.3389/fpls.2020.00796. Gharabadiyan F, Jamali S, Komeili HR (2013) Determination of rook-knot nematode ( Meloidogyne javanica ) damage function for tomato cultivars. J Agr Sci. 58:147-157. https://doi.org/10.2298/JAS1302147G. Hashem M, Abo-Elyousr K (2011) Management of the root-knot nematode Meloidogyne incognita on tomato with combinations of different biocontrol organisms. Crop Protect 30:285-292. https://doi.org/10.1016/j.cropro.2010.12.009. Hooper DU, Solan M, Symstad A, Diaz S, Gessner MO, Buchmann N, Degrange V, Grime P, Hulot F, Mermillod-Blondin F, Roy J, Sephn E, van Peer L (2005) Species diversity, functional diversity, and ecosystem functioning. In: Loreau M, Naeem S, Lnchausti P (Eds), Biodiversity and Ecosystem Functioning: Synthesis and Perspectives, Oxford University Press, pp. 195-281. Hu Y, You J, Wang Y, Long Y, Wang S, Pan F, Yu Z (2022) Biocontrol efficacy of Bacillus velezensis strain YS-AT-DS1 against the root-knot nematode Meloidogyne incognita in tomato plants. Front Microbiol 13:1035748. https://doi.org/10.3389/fmicb.2022.1035748. Hussey RS, Barker KR (1973) A comparison of methods of collecting inocula of Meliodogyne spp., including a new technique. Plant Dis Rep 57:1025-1028. Jamal Q, Cho JY, Moon JH, Munir S, Anees M, Kim KY (2017) Identification for the first time of cyclo (D-Pro-L-Leu) produced by Bacillus amyloliquefaciens y1 as a nematocide for control of Meloidogyne incognita . Molecules 22:1-16. https://doi.org/10.3390/molecules22111839. Jiménez-Aguirre MA, Padilla-Hurtado BE, Ceballos-Aguirre N, Cardona-Agudelo LD, Montoya-Estada CN (2023) Antagonist effect of native bacteria of the genus Bacillus on the root-knot nematode ( Meliodogyne spp.) in tomato germplasm. Chil J Agric Res 83:565-576. http://dx.doi.org/10.4067/s0718-58392023000500565. Jin N, Xue H, Li W, Wang X, Liu Q, Liu S, Liu P, Zhao J, Jian H (2017) Field evaluation of Streptomyces rubrogriseus HDZ-9-47 for biocontrol of Meloidogyne incognita on tomato. J Integ Agric 16:1347–1357. https://doi.org/10.1016/S2095-3119(16)61553-8. Johnson RA, Wichern DW (2007) Applied multivariate statistical analysis, 794. 6 nd ed. Englewood Cliffs, NJ: Prentice Hall. Khan MR, Khan SM, Mohiddin FA, Askary TH (2006) Effects of high nickel soil on root-knot nematode disease of tomato. Nematropica 36:79-87. Khan MR, Mohiddin FA, Ejaz NA, Khan MM (2012) Management of root-knot disease in eggplant through the application of biocontrol bacteria and neem leaves. Turk J Biol 36:161-169. Khan MR, Sharma DN, Ahmad I (2022) Temporal impact of root-knot nematode infection on some important biochemical and physiological characters of tomato. Ind Phytopathol 75:749-758. https://doi.org/10.1007/s42360-022-00500-0. Köberl M (2010) Analyse von Arzneipflanzen-assoziierten Mikroorganismen: Biodiversität und antagonistisches Potenzial gegen bodenbürtige Phytopathogene. MSc, an der Naturwissenschaftlichen Fakultät der Karl-Franzens-Universität Graz, 92p (In Deutsch). Köberl M, Müller M, Ramadan EM, Berg G (2011) Desert farming benefits from microbial potential in arid soils and promotes diversity and plant health. PloS One 6:e24452. https://doi.org/10.1371/journal.pone.0024452. Köberl M, Ramadan EM, Adam M, Cardinale M, Hallmann J, Heuer H, Smalla K, Berg G (2013) Bacillus and Streptomyces were selected as broad-spectrum antagonists against soilborne pathogens from arid areas in Egypt. FEMS Microbiol 342:168-178. https://doi.org/10.1111/1564-6968.12089. Latimer GW (2012) Official Methods of Analysis of AOAC International. 19 th ed, The Association of Official Analysis Chemists International, Maryland, USA. p1399. Lee YS, Kim KY (2016) Antagonistic potential of Bacillus pumilus L1 against root-knot nematode, Meloidogyne arenaria . J Phytopathol 164:29–39. https://doi.org/10.1111/jph.12421 Li X, Hu HJ, Li JY, Wang C, Chen SL, Yan SZ (2019) Effects of the endophytic bacteria Bacillus cereus BCM2 on tomato root exudates and Meloidogyne incognita infection. Plant Dis 103:1551–1558. https://doi.org/10.1094/PDIS-11-18-2016-RE. Lopes-Caitar VS, Pinheiro JB, Marcelino-Guimarães FC (2019) Nematodes in horticulture: An overview. J Hort Sci Crop Res. 1:106. Martinuz A, Schouten A, Sikora RA (2012) Systemically induced resistance and microbial competitive exclusion: Implications on biological control. Phytopathology 102:260- 266. https://doi.org/10.1094/PHYTO-04-11-0120. Mekete T, Mandefro W, Greco N (2003) Relationship between initial population densities of Meloidogyne javanica and damage to pepper and tomato in Ethiopia. Nematol Mediter 31:169‒171. Moran R (1982) Formulae for determination of chlorophyllous pigments extracted with N , N -dimethylformamide. Plant Phys 69:1376–1381. https://doi.org/10.1104/pp.69.6.1376. Mukherjee A, Mondal P, Sinha Babu SP (2020) Nematode extract-induced resistance in tomato against Meloidogyne incognita . Indian J Sci Tech. 13:1476-1479. https://doi.org/10.17485/IJST/v13i14.234. Padilla-Hurtado B, Morillo-Coronado Y, Tarapues S, Burbano S, Soto-Suárez M, Urrea R, Ceballos-Aguirre N (2022) Evaluation of root-knot nematodes ( Meliodogyne spp.) population density for disease resistance screening of tomato germplasm carrying the gene Mi-1 . Chil J Agric Res 82:157-166. https://dx.doi.org/10.4067/S0718-58392022000100157. Pandey SK, Singh H (2011) A simple, cost-effective method for leaf area estimation. J Bot 2011:1–6. https://doi.org/10.1155/2011/658240. Patel VS, Shukla YM, Dhruye JJ (2017) Influence of rrot-knot nematode ( Meliodogyne spp.) on phenolic acid profile in root of tomato ( Solanum lycopersicum L.). Inter J Curr Microb Appl Sci 6:840-848. https://doi.org/10.20546/ijcmas.2017.610.100. Scherwinski K, Grosch R, Berg G (2008) Effect of bacterial antagonists on lettuce: active biocontrol of Rhizoctonia solani and negligible, short-term effects on nontarget microorganisms. FEMS Microbiol Ecol 64:106-116. https://doi.org/10.1111/j.1574-6941.2007.00421.x Seinhorst W (1966) Killing nematodes for taxonomic study with hot F.A.A. Nematol 12:178. Sharma JL, Trevidi PC, Sharma MK, Jiagi B (1990) Alteration in prolin and phenol content of Meloidogyne incognita infected bringal cultivars. Pakist JNematol 8:33–38. Sharma M, Devi S, Chand S (2025) Biocontrol strategies for sustainable management of root-knot nematodes. Physiol Molec Plant Pathol 136:102548. https://doi.org/10.1016/j.pmpp.2024.102548. Sharma M, Jasrotia S, Ohri P, Manhas RK (2019) Nematicidal potential of Streptomyces antibioticus strain M7 against Meloidogyne incognita . AMB Express 9:168. https://doi.org/10.1186/s13568-019-0894-2. Siddiqui ZA, Sharma B, Siddiqui S (2007) Evaluation of Bacillus and Pseudomonas isolated for the biocontrol of Meloidogyne incognita on tomato. Acta Phytopahol Entomol Hungar 42:25-34. https://doi.org/10.1556/APhyto.42.2007.1.4. Stirling GR, Mankau R (1978) Parasitism of Meliodogyne eggs by a new fungal parasite. J Nematol 10:236-240. Taylon AA, Dropkin H, Marin GC (1956) Perineal patterns of the root-knot nematodes. Phytopathology 46:26-34. Teresa Melillo M, Leonetti P, Bongiovanni M, Catagnone-Sereno P, Bleve-Zacheo T (2006) Modulation of reactive oxygen species activities and H 2 O 2 accumulation during compatible and incompatible tomato root-knot nematode interactions. New Phytol 170:501-512. https://doi.org/10.1111/j.1469-8137-2006.01724.x. Viaene N, Coyne DL, Kerry BR (2006) Biological and cultural management. In: Perry RN, Moens M (eds), Plant Nematology, CABI, Wallingford, UK, pp. 346-369. Viljoen JJF, Labuschagne N, Fourie H, Sikora RA (2019) Biological control of the root-knot nematode Meloidogyne incognita on tomatoes and carrots by plant growth-promoting rhizobacteria. Trop Plant Pathol 44:284–291. https://doi.org/10.1007/s40858-019-00283-2. Wickens TD, Keppel G (2004) Design and Analysis: A Researcher’s Handbook. Pearson Prentice-Hall, New Jersey, USA. 624p. Xia Y, Li S, Zhang C, Xu J, Chen Y (2019) B acillus halotolerans strain LYSX1-induced systemic resistance against the root-knot nematode Meloidogyne javanica in tomato. Ann Microbiol 69:1227-1233. https://doi.org/10.1007/s13213-019-01504-4. Xiao L, Wan JW, Yao JH, Feng H, Wei LH (2018) Effects of Bacillus cereus strain Jdm1 on Meloidogyne incognita and the bacterial community in tomato rhizosphere soil. 3 Biotech 8:1–8. https://doi.org/10.1007/s13205-018-1348-2. Yan W, Kang MS (2003) GCE-Biplot Analysis: A Graphical Tool for Breeders, Geneticists, and Agronomists. CRC Press, New York, USA. Yoon GY, Lee YS, Lee SY, Park RD, Hyun HN, Nam Y, Kim KY (2012) Effects on Meloidogyne incognita of chitinase, glucanase and a secondary metabolite from Streptomyces cacaoi GY525. Nematology 14:175-184. https://doi.org/10.1163/138855411X584124. Tables Tables 1 to 7 are available in the Supplementary Files section. Additional Declarations No competing interests reported. Supplementary Files Tables.docx Cite Share Download PDF Status: Published Journal Publication published 13 Jun, 2025 Read the published version in Egyptian Journal of Biological Pest Control → Version 1 posted Editorial decision: Revision requested 15 Apr, 2025 Reviews received at journal 15 Apr, 2025 Reviewers agreed at journal 15 Apr, 2025 Reviewers invited by journal 15 Apr, 2025 Submission checks completed at journal 10 Apr, 2025 First submitted to journal 09 Apr, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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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-6043078","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":443346272,"identity":"648ae366-b101-403c-9ccc-43676393b8e8","order_by":0,"name":"Ahmed MA Mahmoud","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABB0lEQVRIiWNgGAWjYDACCSDmYWCQYWNvgIsZEKWFh43nAKlaGCQSiNTCP7s78cEbBjsePsnHj19XMNglNrA3b5NgqKjFbcmds5sN5zAk87BJp5lZnmFITmzgOVYmwXDmOG5rbuRuk+ZhYAZqSTAzbGA4kNggkWMmwdh2DKcO+Ru523/zMNTzsEke/wbRIv8GqOUfbi0GQFuYeRgO87BJ8Bg/hNjCA9TSUINTi+GN3M2ScwyOAwM5p4yxwSDZuI0nrdgi4dgBnFrkbuRu/PCmolpOvv345o8NFXay/eyHN974UFOH2/sQ54FJNgkQgw3ETGA4TEALBDB/QOIQsmUUjIJRMApGEAAALp5MMJtFZAMAAAAASUVORK5CYII=","orcid":"","institution":"Cairo University","correspondingAuthor":true,"prefix":"","firstName":"Ahmed","middleName":"MA","lastName":"Mahmoud","suffix":""},{"id":443346273,"identity":"419f60ba-5250-4c6f-ad57-647903be532e","order_by":1,"name":"Ahmed ASA El-Eslamboly","email":"","orcid":"","institution":"Agricultural Research Center","correspondingAuthor":false,"prefix":"","firstName":"Ahmed","middleName":"ASA","lastName":"El-Eslamboly","suffix":""},{"id":443346274,"identity":"b5138181-69dc-41cc-bb1e-ae8da12a5126","order_by":2,"name":"Mohamed Adam","email":"","orcid":"","institution":"Cairo University","correspondingAuthor":false,"prefix":"","firstName":"Mohamed","middleName":"","lastName":"Adam","suffix":""},{"id":443346275,"identity":"11706fee-9455-460f-ad15-062dc562cf90","order_by":3,"name":"Mahmoud AA Maraey","email":"","orcid":"","institution":"Cairo University","correspondingAuthor":false,"prefix":"","firstName":"Mahmoud","middleName":"AA","lastName":"Maraey","suffix":""}],"badges":[],"createdAt":"2025-02-16 21:08:15","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6043078/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6043078/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1186/s41938-025-00860-5","type":"published","date":"2025-06-13T15:57:51+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":80783702,"identity":"ae66d6bc-07a1-4b88-8921-35a54bb33f0b","added_by":"auto","created_at":"2025-04-17 05:28:36","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":49868,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eRKN-satisfying response of seed-coated and uncoated tomato ‘Moneymaker’ plants with 25 bacterial antagonists 45 days after RKN-inoculation under greenhouse conditions during the 2019 and 2020 fall seasons. \u003c/strong\u003eBacterial antagonists are presented in Table 1. Data for the number of galls and egg masses per root in both seasons were transformed by the arcsin square root equation. Mean value ± standard error (replicates = 3). According to Duncan’s multiple range test (\u003cem\u003eP\u003c/em\u003e\u0026lt;0.05), the mean values followed by a letter in common are not significant. Vertical bars represent ± standard error (SE) of the mean. The SE was calculated across three replicates for each year. \u003cem\u003eCV\u003c/em\u003e is the coefficient of variation, and the F-value was significant at the 1% probability.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6043078/v1/7929c383ed0176dfec3ce061.png"},{"id":80784534,"identity":"52329fc9-76cd-49a1-a4ce-cd3f04c39906","added_by":"auto","created_at":"2025-04-17 05:36:37","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":42549,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eScree plot of 18 principal components for seed-coated and uncoated tomato 'Moneymaker' plants with 25 bacterial antagonists.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6043078/v1/6870a869bcd505631ecbaba9.png"},{"id":80783709,"identity":"304be0bf-08cd-4051-80e4-d53d8d40a408","added_by":"auto","created_at":"2025-04-17 05:28:36","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":38940,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eBiplot between the first two principal components for 18 estimated traits of seed-coated and uncoated tomato 'Moneymaker' plants with 25 bacterial antagonists.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6043078/v1/94f7220fadaa5aaf6560af7d.png"},{"id":80783705,"identity":"5c33cc66-18e0-4edb-855a-4b6b60248f9f","added_by":"auto","created_at":"2025-04-17 05:28:36","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":23960,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eDendrogram of hierarchical cluster analysis using Euclidean distances on 18 estimated traits of seed-coated and uncoated tomato ‘Moneymaker’ plants with 25 bacterial antagonists.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-6043078/v1/85f551267e2c802a2a329071.png"},{"id":84726652,"identity":"33cf4f55-3cfb-4976-badb-a47054d7b036","added_by":"auto","created_at":"2025-06-16 16:07:46","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1218644,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6043078/v1/3ff19842-ad96-4e52-bc0d-07da4a0bff6e.pdf"},{"id":80783703,"identity":"5d8bc41a-3a2f-4c84-9b14-f24ef1b6ce02","added_by":"auto","created_at":"2025-04-17 05:28:36","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":111000,"visible":true,"origin":"","legend":"","description":"","filename":"Tables.docx","url":"https://assets-eu.researchsquare.com/files/rs-6043078/v1/0aed273ac3e0efd0808d2de3.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Biocontrol of Meloidogyne incognita and Vegetative Growth Stimulation in Tomato ‘Moneymaker’ Plants by Egyptian Soil Bacteria","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eTomato (\u003cem\u003eSolanum lycopersicum\u003c/em\u003e L.) is an important vegetable crop worldwide because of its nutritional and processing value, which is essential for food security and nutrition. The production of tomato worldwide in 2023 was estimated 192.32\u0026nbsp;million tons with an average yield of 35.5 tons hectare\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e across 5.4\u0026nbsp;million hectares(\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://faostat.fao.org/\u003c/span\u003e\u003cspan address=\"https://faostat.fao.org/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). Egypt was the fifth-largest producer worldwide, yielding 6.28\u0026nbsp;million tons from 143,618 hectares in 2022, with an average yield of 43.7 tons hectare\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://faostat.fao.org/\u003c/span\u003e\u003cspan address=\"https://faostat.fao.org/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). Tomato crop is greatly impacted by plant-parasitic nematodes, which can cause yield losses of 30\u0026ndash;40% (Charchar et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2003\u003c/span\u003e), and can even reach 85% in highly susceptible cultivars, particularly in tropical and subtropical regions (Lopes-Caitar et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Tomato production in Egypt is significantly hindered by the presence of root-knot nematodes (RKN; \u003cem\u003eMeloidogyne\u003c/em\u003e spp.), particularly \u003cem\u003eM. incognita\u003c/em\u003e (Abd-Algawad \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eNematode control is a complex and costly process with limited options. Integrated pest management (IPM) for RKN in sustainable agriculture systems involve four main strategies: host plant resistance, agricultural practices, biological control, and chemical control. Chemical nematicides, such as methyl bromide, chloropicrin, 1,3-dichloropropene, or metam-based products (metam sodium and metam potassium), effectively reduce RKN damage, but their cost and potential harmful impacts on human health and the environment often make them less appealing to farmers (Castellano-Hinojosa et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). IPM is now increasingly focused on biological control, which uses antagonistic organisms to reduce nematode populations and their abundance and damage, providing a sustainable alternative to chemical nematicides. Plant-parasitic nematodes can be biocontrolled by using a variety of antagonistic organisms, known as biological control agents (BCAs), including fungi, bacteria, viruses, protists, nematode antagonists, and other invertebrates (Sharma et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). BCAs can directly control RKN through resource competition, predation, and parasitism at various life stages, or indirectly by producing nematicidal metabolites and inducing plant resistance (Sharma et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). The use of BCAs in suppressing nematode populations has been controversial because other soil microorganisms and the host plant can be adversely affected (Sharma et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Furthermore, beneficial plant\u0026ndash;microorganism interactions are commonly exact, with only a few broad-spectrum antagonists identified (Sharma et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2025\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cem\u003eMeloidogyne\u003c/em\u003e spp, can be suppressed by indigenous microbial communities in arable soils (Adam et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2014\u003c/span\u003e, Amorim et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2024\u003c/span\u003e, Sharma et al., \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Identifying antagonistic microbes and understanding the mechanisms that regulate nematode populations is particularly important in soils that naturally suppress \u003cem\u003eMeloidogyne\u003c/em\u003e spp. Rhizosphere bacterial genera such as \u003cem\u003eBacillus\u003c/em\u003e, \u003cem\u003ePaenibacillus\u003c/em\u003e, \u003cem\u003ePseudomonas\u003c/em\u003e, and \u003cem\u003eStreptomyces\u003c/em\u003e, have been successfully used as BCAs for nematodes in vegetable crops, including tomato (Amorim et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2024\u003c/span\u003e, Sharma et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2025\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eRhizosphere bacteria, particularly \u003cem\u003eBacillus\u003c/em\u003e species, have shown the ability to control \u003cem\u003eMeloidogyne\u003c/em\u003e spp. in tomato (Hu et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2022\u003c/span\u003e, Jim\u0026eacute;nez-Aguirre et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Species such as \u003cem\u003eB. megaterium\u003c/em\u003e, \u003cem\u003eB. circulans\u003c/em\u003e, \u003cem\u003eB. subtilis\u003c/em\u003e, \u003cem\u003eB. firmus\u003c/em\u003e, and \u003cem\u003eB. pumilus\u003c/em\u003e produce enzymes (d\u0026rsquo;Errico et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2019\u003c/span\u003e, Lee et al. 2016, Viaene et al. \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2006\u003c/span\u003e) and bioactive compounds (Ayaz et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2021\u003c/span\u003e, Jamal et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) that damage nematodes and inhibit their development, as well as promote plant growth by activating systemic resistance (Adam et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2014\u003c/span\u003e, Xia et al. \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) and regulating defense-related genes in tomato plants (Xia et al. \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). The behavior and feeding patterns of nematodes can also be modified by these bacteria (Viaene et al. \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). Strains of \u003cem\u003eB. aryabhattai\u003c/em\u003e, \u003cem\u003eB. cereus\u003c/em\u003e, \u003cem\u003eB. firmus\u003c/em\u003e, \u003cem\u003eB. halotolerans\u003c/em\u003e, \u003cem\u003eB. pumilus\u003c/em\u003e, \u003cem\u003eB. subtilis\u003c/em\u003e, and \u003cem\u003eB. velezensis\u003c/em\u003e have been shown to reduce nematode populations, prevent gall formation, and inhibit egg hatching both in vitro and in vivo (Amorim et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2024\u003c/span\u003e, Hu et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2022\u003c/span\u003e, Khan et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2012\u003c/span\u003e, Viljoen et al. \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2019\u003c/span\u003e, Xia et al. \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2019\u003c/span\u003e, Xiao et al. \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). The reproductive cycle of \u003cem\u003eM. incognita\u003c/em\u003e and paralysis in nematode juveniles can be disrupted by \u003cem\u003eB. cereus\u003c/em\u003e (Colagiero et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2018\u003c/span\u003e, Li et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2019\u003c/span\u003e, Xiao et al., \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Systemic resistance is induced by other strains of \u003cem\u003eB. halotolerans\u003c/em\u003e and \u003cem\u003eB. atrophaeus\u003c/em\u003e by producing volatile organic compounds (VOCs) that have nematocidal properties (Ayaz et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Recent research has validated that \u003cem\u003eB. cereus\u003c/em\u003e IBCBb 116 significantly reduces nematode populations and enhances tomato plant growth, with the potential for IPM, especially when combined with susceptible tomato genotypes (Amorim et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cem\u003ePseudomonas\u003c/em\u003e species, particularly \u003cem\u003eP. aeruginosa\u003c/em\u003e and \u003cem\u003eP. fluorescens\u003c/em\u003e, have shown strong potential in controlling RKN (\u003cem\u003eM. javanica\u003c/em\u003e and \u003cem\u003eM. incognita\u003c/em\u003e) in humid soils (Colagiero et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2018\u003c/span\u003e, Hashem and Abo-Elyousr \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2011\u003c/span\u003e, Khan et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). \u003cem\u003eP. aeruginosa\u003c/em\u003e isolates can inhibit the growth of \u003cem\u003eM. javanica\u003c/em\u003e and suppress their populations, as well as prevent root galling by producing hydrogen cyanide (Stirling and Mankau \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e1978\u003c/span\u003e, Siddiqui et al. \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). \u003cem\u003eP. fluorescens\u003c/em\u003e enhanced the defense enzyme activity in RKN-resistant tomato plants and produced antibiotics and enzymes that inhibit nematode egg hatching and reduce egg mass (Khan et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). \u003cem\u003eP. aeruginosa\u003c/em\u003e IBCBb 122 showed over 68% mortality in \u003cem\u003eM. incognita\u003c/em\u003e juveniles in vitro and effectively reduced nematode populations in greenhouse tomato pots (Amorim et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Nasima et al. (2002) found that \u003cem\u003ePseudomonas\u003c/em\u003e sp. filtrates can increase juvenile mortality, decrease nematode populations, and improve tomato plant growth and yield. Similarly, \u003cem\u003ePaenibacillus\u003c/em\u003e species, like \u003cem\u003eP. polymyxa\u003c/em\u003e, \u003cem\u003eP. barcinonensis\u003c/em\u003e, and \u003cem\u003eP.alvei\u003c/em\u003e, have demonstrated nematicidal effects. \u003cem\u003eP. polymyxa\u003c/em\u003e KM2501-1 eliminated \u003cem\u003eM.\u003c/em\u003e incognita through contact or fumigation in honey traps (Cheng et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), while \u003cem\u003eP. barcinonensis\u003c/em\u003e A10 and \u003cem\u003eP. alvei\u003c/em\u003e T30 caused paralysis in \u003cem\u003eM. incognita\u003c/em\u003e J\u003csub\u003e2\u003c/sub\u003e juveniles (Viljoen et al. \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Actinobacteria, especially \u003cem\u003eStreptomyces\u003c/em\u003e spp., areabundant in soil and have been demonstaretd to producednematicidal compounds such as actinomycins and chitinase, that directly target nematodes (Jin et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2017\u003c/span\u003e, Yoon et al. \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2012\u003c/span\u003e, Sharma et al. \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Species such as \u003cem\u003eS. albogriseolus\u003c/em\u003e, \u003cem\u003eS. cacaoi\u003c/em\u003e, \u003cem\u003eS. antibioticus\u003c/em\u003e, and \u003cem\u003eS. rubrogriseus\u003c/em\u003e have been proven through parasitism and toxins production (Jin et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2017\u003c/span\u003e, Yoon et al. \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). Actinobacteria also contribute to naturally suppressive soils by anhancing plant growth, suppressing pathogens, and strengthening plant defenses.\u003c/p\u003e \u003cp\u003eAntagonistic bacteria that can control nematodes and soil-borne pathogens have been spotted in Egyptian organic farming soils and on the roots of medicinal plants and have been isolated and evaluated for effectiveness. Various bacterial species/isolates showed antagonism against nematodes \u003cem\u003ein vitro\u003c/em\u003e studies by K\u0026ouml;berl et al. (2010, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2011\u003c/span\u003e, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2013\u003c/span\u003e) and Adam et al. (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Based on this, this study aimed to evaluate 20 bacterial isolates from the genera \u003cem\u003eBacillus\u003c/em\u003e, \u003cem\u003eLysobacter\u003c/em\u003e, \u003cem\u003ePaenibacillus\u003c/em\u003e, and \u003cem\u003eStreptomyces\u003c/em\u003e for their potential in biocontrol RKN and promot growth in the susceptible tomato \u0026lsquo;Moneymaker\u0026rsquo; under \u003cem\u003ein vivo\u003c/em\u003e conditions. Their performance was compared to known effective microbial biological agents: \u003cem\u003eBacillus subtilis\u003c/em\u003e subsp. \u003cem\u003esubtilis\u003c/em\u003e Sb1-20, \u003cem\u003ePseudomonas trivialis\u003c/em\u003e 3Re2-7, \u003cem\u003eRhizobium etli\u003c/em\u003e G12, and \u003cem\u003eSporosarcina psychrophile\u003c/em\u003e Sd4-11.\u003c/p\u003e"},{"header":"2. MATERIALS AND METHODS","content":"\u003cp\u003e\u003cstrong\u003e2.1. Nematode culture\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA pure culture of RKN was established by using naturally infected tomato roots. The \u003cem\u003eMeloidogyne\u003c/em\u003e species was determined using the perineal pattern system for mature females as described by Taylon et al. (1956) and Seinhorst (1966). An egg mass from a female of \u003cem\u003eM. incognita\u003c/em\u003e was used to inoculate eggplant plants using a perennial pattern. Eggs were extracted from heavily galled roots using a 1.5% chlorine solution in order to prepare nematode inoculum according to Hussey and Barker (1973). Roots were cut, put in a bottle with solution, and shaken strongly for 3 min. The egg suspension was received on the 20 \u0026micro;m sieve, then placed on a modified Baermann dish and incubated at 25\u0026plusmn;2\u0026deg;C for 7-10 days (Hooper et al. 2005). Hatched J2s were collected daily and stored at 6\u0026deg;C to be used in the experiments.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.2. Plant materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSusceptible tomato LA2706 \u0026lsquo;Moneymaker\u0026rsquo; (\u003cem\u003emi-1\u003c/em\u003e/\u003cem\u003emi-1\u003c/em\u003e) (https://tgrc.ucdavis.edu/) was used in this study. Tomato \u0026apos;Moneymaker\u0026apos; plants were cultivated in a greenhouse during the 2019 winter season for seed propagation.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.3. Bacterial antagonists\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTwenty-four bacterial isolates of various genera, listed in Table 1, were evaluated for their potential to RKN-suppress and stimulate vegetative growth of tomato \u0026lsquo;Moneymaker\u0026rsquo; plants, with \u003cem\u003eEscherichia coli\u003c/em\u003e JM109 as the negative control. Bacterial isolates included 20 bacterial isolates (B1-B20) that were previously tested in-vitro for RKN suppression of \u003cem\u003eM. incognita\u003c/em\u003e by K\u0026ouml;berl et al. (2013), and four bacterial isolates (B21 - B24) have previously shown activity against RKN and soil-borne fungal pathogens (K\u0026ouml;berl et al. 2010, 2011, Martinuz et al. 2012, Scherwinski et al. 2008) as positive controls. The bacterial antagonists were cultured on Luria-Bertani (LB) medium in Petri dishes 9 cm in diameter at 27\u0026plusmn;3 \u0026deg;C and 16/8h photoperiod for 3 days until complete growth on the medium surface.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.4. Experimental procedure\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSeeds were thoroughly sterilized in 2% sodium hypochlorite (half strength of commercial bleach) for 10 min on the shaker, washed four times with distilled water, and placed on filter paper to dry under sterile conditions. Fifty seeds were mixed in a bacterial lawn until the bacteria completely coated the seed surface. The uncoated seeds were considered another negative control. The coated seeds were left for a few minutes under a laminar flow hood to dry. The coated and uncoated seeds were sown on September 1\u003csup\u003est\u0026nbsp;\u003c/sup\u003ein the 2019 and 2020 seasons in 209-cell seedling trays filled with a mixture of coconut peat and vermiculite (volume 1:1) enriched with macro and microelements. Seeds were seeded into trays with two empty rows between each coating treatment to prevent cross-contamination of bacterial isolates. The seeds were planted under saran-house conditions at the Department of Agricultural Zoology and Nematology, Faculty of Agriculture, Cairo University (30\u0026deg;00\u0026apos;59.8\u0026quot;N 31\u0026deg;12\u0026apos;21.0\u0026quot;E). Three-week-old seedlings were transplanted into a plastic pot with a 15 cm diameter (seedling/pot), containing 600 g of sterilized a clay: sandy mixture (1:1 volume), and watered to field\u0026rsquo;s capacity. Pots were arranged in a randomized complete block design (RCBD) with three replicates under saran-house conditions. Each experimental unit contained ten plants. Plants were subjected to common agricultural practices without applying pesticides. After a week, every pot was inoculated with 1,500 freshly hatched J2 in four holes of 2 cm depth at 3 cm distance from the stem base.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.5. Efficacy of the bacterial isolates on RKN-satisfying response and growth improvement of tomato\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe traits related to the RKN-satisfying response, vegetative growth, and metabolic activity of \u0026lsquo;Moneymaker\u0026rsquo; plants from various seed-coating treatments were assessed 45 days after inoculation (DAI).\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cstrong\u003e2.5.1. The RKN-satisfying response\u003c/strong\u003e\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eBanora and Almaghrabi (2019) states that the RKN-satisfying response involves the plant\u0026rsquo;s ability to suppress the development or reproduction of RKN compared to a susceptible plant of the same species (Banora and Almaghrabi 2019). The RKN-satisfying responses of tomato \u0026lsquo;Moneymaker\u0026rsquo; plants grown with coated and uncoated seeds were assessed. Nematode populations of each plant\u0026rsquo;s root system were extracted using the method described by Hooper et al. (2005). The response was evaluated by counting the number of egg masses (NEM) and galls (NG) in a root system.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cstrong\u003e2.5.2. Plant vegetative growth and metabolic activity\u003c/strong\u003e\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003ePlant measurements were estimated based on the five most representative plants in each EU. Vegetative growth traits included stem length (SL), stem diameter (SD), number of plant leaves (NPL), the area of the fifth fully expanded leaf from the top of the plant (LA), fresh and dry weights of shoots (FSW \u0026amp; DSW) and roots (RFW \u0026amp; RDW), and leaf content of photosynthesis pigments. SL was measured from the soil surface to the stem tip, while SD was measured about 1cm above the soil. The leaf area was measured using the leaf-weighting technique (Pandey and Singh 2011).\u0026nbsp;The photosynthesis pigments were extracted by grinding 0.5g of fresh leaves with 5ml of dimethylformamide according to Moran (1982). Leaf photosynthesis pigments (chlorophyll a: chlor-a, chlorophyll b: chlor-b, total chlorophyll: t-chlor, and total carotenoids: t-carot) were measured using a Jenway 6305 UV/visible spectrophotometer (Jenway, Wales, UK) at 664, 647, and 480 nm (Moran 1982). The formulas for calculating pigment concentrations were as follows: chlor-a = 11.65A\u003csub\u003e664\u003c/sub\u003e \u0026ndash; 2.69A\u003csub\u003e647\u003c/sub\u003e, chlor-b = 20.81A\u003csub\u003e647\u003c/sub\u003e \u0026ndash; 4.53A\u003csub\u003e664\u003c/sub\u003e, and t-carot = (1000A\u003csub\u003e480\u003c/sub\u003e \u0026ndash; 1.42Ca \u0026ndash; 46.09Cb)/202.\u003c/p\u003e\n\u003cp\u003e100g of fresh weight of each shoot and root was oven-dried at 70\u0026ordm;C until a constant weight was reached to measure the dry matter content of shoots (SDM) and roots (RDM). The AOAC method was used to estimate the root content of carbohydrates (Carb), proteins (Pr), and polyphenols (PPh)\u0026nbsp;in dry weight (Latimer 2012).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.6. Statistical Analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe Shapiro-Wilk test was performed to check the normality of the data, revealing significant differences in NG, NEM, SL, SFW, RDM, Chlor-a, Chlor-b, Carb, Pr, and PPh in both seasons; NPL, SDW, and RDW in the 2019 season only; and T-chlor and T-Carot in the 2020 season only. Consequently, an arcsine square root transformation was applied to these variables for statistical analysis (Wickens and Keppel 2004). For each season, the variability in the effectiveness of bacterial isolates in suppressing RKN and enhancing vegetative growth of \u0026lsquo;Moneymaker\u0026rsquo; tomato plants was assessed using variance components derived from RCBD-ANOVA and the coefficient of variation (\u003cem\u003eCV\u0026nbsp;\u003c/em\u003e= standard deviation / mean). The CV are classified as low (\u0026lt;10%), moderate (10-20%), high (20-30%), and very high (\u0026gt;30%) as mentioned by \u003cstrong\u003eBeah et al. (2021)\u003c/strong\u003e.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eA high coefficient of variation indicates significant influence by bacterial isolates. MSTATc v.2.1 software (Michigan State University, Michigan, USA) was used to perform these analyses.\u003c/p\u003e\n\u003cp\u003ePrincipal component analysis (PCA) and hierarchical cluster analysis (HCA) were used to evaluate variation and similarity/dissimilarity among coating and uncoating seed plants. The data from both seasons were analyzed was combined and analyzed. The data were standardized with Z-scores to account for scale differences before analysis. The latent root criterion (eigenvalue \u0026gt;1) and parallel analysis were used to determine significant components using varimax rotation in PCA (Johnson and Wichern 2007). To assess correlations between traits and the relationships between treatments and traits, the biplot of the top two PCs was constructed according to Yan and Kang (2003). The cosine of the angle between the vectors was used to estimate correlations: an angle of 90\u0026deg; indicates no correlation, \u0026lt;90\u0026deg; indicates a positive correlation, and \u0026gt;90\u0026deg; indicates a negative correlation. Furthermore, Pearson\u0026rsquo;s correlation coefficients were used to identify traits that are directly related to RKN-satisfying responses in both coated and uncoated-seed tomato \u0026lsquo;Moneymaker\u0026rsquo; plants. HCA was performed using the Euclidean distance and unweighted pair-group average (UPGA) method. Multivariate analyses including Pearson correlation were performed using the IBM SPSS software version 26.0.0 (SPSS Inc., Chicago, Illinois, USA) and XLSTAT software version 2019 (Addinsoft, Paris, France).\u003c/p\u003e"},{"header":"3. RESULTS AND DISCUSSION","content":"\u003cp\u003e\u003cstrong\u003e3.1. RKN-satisfying response\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn both seasons, seed \u0026lsquo;Moneymaker\u0026rsquo; coating with several bacterial antagonists had a highly significant effect (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.001) on the RKN-satisfying responses, i.e., NG (966.826\u003csup\u003e***\u003c/sup\u003e and 342.406\u003csup\u003e***\u003c/sup\u003e, respectively) and NEM (921.924\u003csup\u003e***\u003c/sup\u003e and 488.081\u003csup\u003e***\u003c/sup\u003e, respectively), of their plants in both seasons (Figure 1). This was further supported by the notably very high CV values for both NG (57.6 and 53.8%, respectively) and NEM (56.5 and 65.3%, respectively) in each season (Figure 1). NG on the roots of \u0026lsquo;Moneymaker\u0026rsquo; plants ranged from 15.68 with B23-coated seed plants to 60.51 with uncoated seed plants in 2019, and from 15.00 with B23-coated seed plants to 44.50 with uncoated seed plants in 2020 (Figure 1). The highest NG in both seasons was found with uncoated seed plants (60.51 \u0026amp; 44.50, respectively) and B11-coated seed plants (60.11 \u0026amp; 38.00, respectively), with significant similarity (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) among them in 2019 only. The lowest NG in both seasons were with coated seed plants in B23 (15.68 \u0026amp; 15.00, respectively) and B19 (21.27 \u0026amp; 16.00, respectively), with significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) among them in 2020 only.\u003c/p\u003e\n\u003cp\u003eNEM per root of \u0026lsquo;Moneymaker\u0026rsquo; plants ranged from 18.26 with B23-coated seed plants to 61.74 with B11-coated seed plants in 2019 and from 15.33 with B23-coated seed plants to 68.00 with uncoated plants in 2020 (Figure 1). The uncoated seed plants had the highest NEM during both seasons (62.13 \u0026amp; 68.00, respectively) with significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) with coated seed plants by B5, B11, and B25 in 2019 only (61.22, 61.74, and 61.03, respectively). B23-coated seed plants showed the lowest NEM in both seasons (18.26 \u0026amp; 15.33, respectively), significantly similar to B19-coated seed plants in 2020 (16.50). The B22-coated plants were ranked in the next significantly lowest NEM (23.51 \u0026amp; 18.00, respectively), followed by coated seed plants by B13 (26.76\u0026amp;19.75, respectively) and B24 (26.56 \u0026amp; 19.00, respectively).\u003c/p\u003e\n\u003cp\u003eSeed coating with bacterial antagonists enhanced the resistance of \u0026lsquo;Moneymaker\u0026rsquo; plants to RKN, showing a reduced RKN-satisfying response in coated seed plants compared to uncoated ones (Figure 1). The presence of bacterial strains of \u003cem\u003eBacillus\u003c/em\u003e, \u003cem\u003ePaenibacillus\u003c/em\u003e, and \u003cem\u003ePseudomonas\u003c/em\u003e in tomato can result in systematic resistance to \u003cem\u003eMeloidogyne\u003c/em\u003e spp. (Siddiqui et al. 2007, Adam et al. 2014, Xia et al. 2019, Ghahremani et al. 2020). \u003cem\u003eB. subtilis\u0026nbsp;\u003c/em\u003eSb4-23, Mc5-Re2, and Mc2-Re2 (Adam et al. 2014), and \u003cem\u003eB. velezensis\u003c/em\u003e YS-AT-DS1 (Hu et al. 2022) can reduced \u003cem\u003eM. incognita\u0026nbsp;\u003c/em\u003eJ2 infection and related damage, including NG and NEMs. \u0026nbsp;The inhibition of gall formation, egg hatching, and J2 activity of \u003cem\u003eM. incognita\u003c/em\u003e was found to be mediated by \u003cem\u003eB. cereus\u003c/em\u003e Jdm1, according to Xiao et al. (2018) findings. Colagiero et al. (2018) showed that \u003cem\u003eB. cereus\u003c/em\u003e, \u003cem\u003eB. licheniformis\u003c/em\u003e, \u003cem\u003ePaenibacillus fluoresencs,\u0026nbsp;\u003c/em\u003eand \u003cem\u003eP. brassicacearum\u0026nbsp;\u003c/em\u003edisrupted the reproductive cycle of \u003cem\u003eM. incognita\u003c/em\u003e. The presence of \u003cem\u003eB. cereus\u003c/em\u003e BCM2 prevented \u003cem\u003eM. incognita\u0026nbsp;\u003c/em\u003ejuveniles from reaching tomato plants, as reported by Li et al. (2019). According to Viljoen et al. (2019), \u003cem\u003eB. firmus\u003c/em\u003e T11, \u003cem\u003eB. aryabhattai\u003c/em\u003e A08, \u003cem\u003ePaenibacillus barcinonensis\u003c/em\u003e A10, \u003cem\u003eP. alvei\u003c/em\u003e T30, and \u003cem\u003eB. cereus\u003c/em\u003e have paralyzed \u003cem\u003eM. incognita\u0026nbsp;\u003c/em\u003eJ2\u003cem\u003e.\u003c/em\u003e According to Ghahremani et al. (2020), \u003cem\u003eB. firmus\u003c/em\u003e I-1582 was responsible for killing \u003cem\u003eMeloidogyne\u0026nbsp;\u003c/em\u003espp. nematodes and prevented their egg from hatching. Ayaz et al. (2021) found that \u003cem\u003eB. atrophaeus\u003c/em\u003e GBSC56 produced VOCs, which induced severe oxidative stress in \u003cem\u003eM. incognita\u003c/em\u003e and enhanced the activity of antioxidant enzymes in infested tomato roots.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3.2. Vegetative growth performance\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eRKN infection in tomato plants causes galls to form in the root system, disrupting nutrient and water uptake. These galls also create feeding sites in the vascular tissues, which harbor adult female nematodes. This results in symptoms of yellowing leaves, wilting plants and stunting their growth, floral abortions, reduced fruit quantity and quality, and, in severe cases, plant death (Banora 2023). Evaluating vegetative growth traits is an important indicator of plant resistance to RKN.\u003c/p\u003e\n\u003cp\u003eThe vegetative growth performance of tomato \u0026lsquo;Moneymaker\u0026rsquo; coated and uncoated seed plants at 45 DAI under RKN infestation is presented in Table 2. Seed \u0026lsquo;Moneymaker\u0026rsquo; coating with several bacterial antagonists had a highly significant effect (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.001) on the plant\u0026rsquo;s vegetative growth performance under RKN infestation conditions in both seasons (Table 2). This was further confirmed by the relatively moderate to very high CV observed for vegetative traits in each season, ranging 16.45-60.37% in 2019 and 10.94-51.28% in 2020 (Table 2).\u003c/p\u003e\n\u003cp\u003eSL (cm) of \u0026lsquo;Moneymaker\u0026rsquo; plants ranged from 30.67 with uncoated seed to 109.03 with coated seed plants by B16 and B17 in 2019, and from 50.00 with uncoated seed plants to 74.72 with coated seed plants by B6 and B16 in 2020. The uncoated seed plants had the shortest stems in both seasons (30.67 \u0026amp; 50.00, respectively), with significant similarities with coated seed plants by B1, B8, B10, B11, B18, and B22 in 2019 (ranging from 30.70 - 53.68). B16-coated seed plants had the tallest plants in both seasons (109.03 \u0026amp; 74.72, respectively), with significant similarities (\u003cem\u003eP\u0026nbsp;\u003c/em\u003e\u0026lt;0.05) with coated plants by B4, B14, B15, B17, B21, and B24 in 2019 (ranging from 82.92 \u0026ndash; 109.03) and those by B6, B9, and B20 in 2020 (74.72, 73.13, and 73.17 cm, respectively).\u003c/p\u003e\n\u003cp\u003eSD (mm) of \u0026lsquo;Moneymaker\u0026rsquo; plants ranged from 2.40 with uncoated plants to 5.13 with B19-coated seed plants in 2019 and from 4.52 with B9-coated seed plants to 5.80 with B19-coated seed plants in 2020 (Table 2). The uncoated seed plants have the thinnest stems in both seasons (2.40 \u0026amp; 4.70, respectively), with significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) with coated seed plants by B2, B9, B15, and B16 in 2020 only (4.63, 4.52, 4.62, and 4.75, respectively). B19-coated seed plants had the thickest stems in both seasons (5.13 \u0026amp; 5.80, respectively), with significant similarity (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) with those coated by B9 in 2019 only (5.07), and B11 and B14 in 2020 (5.57 \u0026amp; 5.65, respectively).\u003c/p\u003e\n\u003cp\u003eNPL of \u0026lsquo;Moneymaker\u0026rsquo; plants ranged from 5.0 with uncoated seed plants and B1-coated seed plants to 12.4 with B14-coated plants in 2019, and from 7.17 with B12-coated seed plants to 11 with B22-coated seed plants in 2020 (Table 2). The lowest NPL in both seasons was found with uncoated seed plants (5.0 and 8.0, respectively) and B1-coated seed plants (5.0 \u0026amp; 8.0, respectively), with significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) among them and coated seed plants by B2, B9, B11, B15, B16, B19, and B25 in 2020 (ranging from 8.17 \u0026ndash; 8.67). B4-coated seed plants had the most leaves in both seasons (12.40 \u0026amp; 10.83, respectively), with significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) with coated seed plants by B4, B14, and B17 in 2019 (12.0, 12.40, and 12.0, respectively), and those by B22 in 2020 (11.0).\u003c/p\u003e\n\u003cp\u003eSFW (g) of \u0026lsquo;Moneymaker\u0026rsquo; plants ranged from 28.13 with uncoated seed plants to 55.98 with B23-coated seed plants in 2019, and from 34.08 with uncoated seed plants to 61.00 with B23-coated seed plants (Table 2). The uncoated seed plants had the lowest SFW in both seasons (28.13 \u0026amp; 34.08, respectively), followed by coated seed plants by B12 (29.92 \u0026amp; 35.93, respectively), B15 (29.57 \u0026amp; 35.67, respectively), and B22 (29.40 \u0026amp; 35.50, respectively), with significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) among them in only 2020, and coated seed plants by B1, B3, B7, B8, B13, B14, B21, and B25 in 2020 (ranging from 36.83 \u0026ndash; 38.12). B23-coated seed plants had the highest SFW in both seasons (55.98 \u0026amp; 61.00, respectively), followed by B17-coated seed plants (49.20\u0026amp;55.55, respectively), and then B18-coated seed plants (45.55 \u0026amp; 51.82, respectively). SDW (g) of \u0026lsquo;Moneymaker\u0026rsquo; plants ranged from 3.49 with uncoated seed plants to 7.06 with B23-coated seed plants in 2019, and from 4.01 with uncoated seed plants to 8.21 with B23-coated seed plants (Table 2). The uncoated seed plants had the lowest SDW in both seasons (3.49\u0026amp;4.01, respectively), followed by coated seed plants by B1 (3.65 \u0026amp; 4.02, respectively) and B22 (3.74 \u0026amp; 4.06, respectively), with significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) among them in 2020, and coated seed plants by B7 and B25 in 2020 (4.00 \u0026amp; 4.61, respectively). In both seasons, B23-coated seed plants had the highest SDW (7.06 \u0026amp; 8.21, respectively), followed by B17-coated seed plants (6.21 \u0026amp; 6.92, respectively).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eRFW (g) ranged from 1.80 with uncoated seed plants to 9.23 with B20-coated seed plants in 2019, and from 3.03 with uncoated seed plants to 10.46 with B20-coated seed plants in 2020 (Table 2). The uncoated seed plants had the lowest RFW in both seasons (1.80 \u0026amp; 3.03, respectively), with significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) with coated seed plants by B1, B3, B13, B21, and B25 in only 2020 (4.25, 4.27, 3.59, 4.59, and 4.43, respectively). In both seasons, B20-coated seed plants had the highest RFW (9.23\u0026amp;10.46, respectively), followed by coated seed plants by B12 (7.18 \u0026amp; 8.41, respectively), and then B19 (6.93 \u0026amp; 8.16, respectively). RDW (g) ranged from 0.40 with uncoated seed plants to 1.97 with B20-coated seed plants in 2019, and from 0.68 with uncoated seed plants to 2.31 with B20-coated seed plants in 2020 (Table 2). The lowest RDW in both seasons was found with uncoated seed plants (0.40 \u0026amp; 0.68, respectively) and B13-coated seed plants (0.42 \u0026amp; 0.70 g, respectively), with significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) among them and coated seed plants by B1, B3, B7, and B25 in 2020 (0.91, 0.92, 0.95, and 0.97, respectively). In both seasons, B20-coated seed plants had the highest RDW (1.97 \u0026amp; 2.31, respectively), followed by coated seed plants by B12 (1.52 \u0026amp; 1.56, respectively) and B15 (1.49 \u0026amp; 1.64, respectively), then coated seed plants by B10 (1.41 \u0026amp; 1.79, respectively), B11 (1.43 \u0026amp; 1.56, respectively), and B18 (1.44 \u0026amp; 1.79, respectively).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3.3. Foliar photosynthesis pigments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe foliar photosynthesis pigment content of tomato \u0026lsquo;Moneymaker\u0026rsquo; coated and uncoated seed plants is presented in Table 3. Seed \u0026lsquo;Moneymaker\u0026rsquo; coating with several bacterial antagonists had a highly significant effect (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.001) on the leaf content of photosynthesis pigments under RKN infestation conditions in both seasons (Table 3). Further support was provided by the consistently high to very high CV values recorded for the leaf content of photosynthesis pigments across seasons, which ranged from 27.07-120.19% in 2019 and from 23.49-32.57% in 2020 (Table 3).\u003c/p\u003e\n\u003cp\u003eThe leaf chlor-a content (mg g\u003csup\u003e-1\u003c/sup\u003e) ranged from 0.58 with B11-coated seed plants to 2.13 with B14-coated seed plants in 2019, and from 0.78 with B13-coated seed plants to 1.54 with B21-coated seed plants in 2020 (Table 3). Leaf chlor-a content was inconsistent during both seasons. The highest leaf chlor-a content was found in B14-coated seed plants in 2019 (2.13), while in coated seed plants by B21 and B14 in 2020 (1.54 \u0026amp; 1.53, respectively) with significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) among them. In 209, the lowest leaf chlor-a content was found in 2019 with coated seed plants by B1, B2, B10, B11, and B12 (ranging from 0.58 - 0.79), with significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) among them. In 2020, uncoated seed plants and coated seed plants by B4, B13, B17, B18, and B24 had the lowest leaf chlor-a content (ranging from 0.78 - 0.89).\u003c/p\u003e\n\u003cp\u003eLeaf chlor-b content (mg g\u003csup\u003e-1\u003c/sup\u003e) ranged from 0.05 with uncoated seed plants to 0.81 with B23-coated seed plants in 2019, and from 0.26 with B6-coated seed plants to 0.46 with B23-coated seed plants in 2020 (Table 3). B23-coated seed plants had the highest leaf chlor-b content in both seasons (0.81 \u0026amp; 0.46, respectively), with significant similarity (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) with B21-coated seed plants in 2020 (Table 3). Uncoated seed plants had the lowest leaf chlor-b content in both seasons (0.05 \u0026amp; 0.28, respectively), with significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) with seed-coated seed plants by B10 and B20 in 2019 (0.07 \u0026amp; 0.10, respectively), and coated seed plants by B4, B6, B9, B13, B17, B18, and B24 in 2020 (ranging from 0.26 \u0026ndash; 0.28).\u003c/p\u003e\n\u003cp\u003eLeaf total t-chlor content (mg g\u003csup\u003e-1\u003c/sup\u003e) was between 0.73 with B10-coated seed plants and 2.26 with B14-coated seed plants in 2019, and between 1.06 with B13-coated seed plants and 1.95 with B21-coated seed plants in 2020 (Table 3). Leaf t-chlor content was inconsistent in both seasons, particularly the highest content. The highest content was found in B14-coated seed plants in 2019 (2.26), and in coated seed plants by B21 and B23 (1.95 \u0026amp; 1.94, respectively) in 2020. The uncoated seed plants had the lowest t-chlor content in both seasons (0.76 \u0026amp; 1.10, respectively), with significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) with coated seed plants by B2, B10, and B11 in 2019 (0.83, 0.73, and 0.76, respectively), and coated seed plant by B4, B17, B18, B13, and B24 in 2020 (ranging from 1.06 - 1.16).\u003c/p\u003e\n\u003cp\u003eLeaf t-car (mg g\u003csup\u003e-1\u003c/sup\u003e) was between 0.40 with uncoated seed plants to 0.75 with coated seed plants by B1 and B17 in 2019, and between 0.65 with B24-coated seed plants to 1.05 with B11-coated seed plants in 2020 (Table 3). B11-coated seed plants had the highest leaf t-car content in both seasons (0.71 \u0026amp; 1.05, respectively), with significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) with coated seed plants by B1 and B17 in 2019 (0.75 for both), and coated seed plants by B21 in 2020 (1.04) (Table 3). The uncoated seed plants had the lowest leaf t-car content in both seasons (0.40 \u0026amp; 0.67, respectively), with significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) with B21-coated seed plants in 2019 (0.43), and with coated seed plants by B23 and B24 in 2020 (0.67 \u0026amp; 0.65, respectively).\u003c/p\u003e\n\u003cp\u003eSeed-coating tomato \u0026lsquo;Moneymaker\u0026rsquo; plants with some bacterial antagonists improved vegetative growth under RKN infestation. The vegetative growth performance of uncoated seed plants was the lowest, while coated seed plants showed enhanced vegetative growth, with some bacterial isolates yielding better results. Coated seed plants by B5, B12, B19, B20, B22, and B23 exhibited favorable vegetative growth traits. The RKN-satisfying response of tomato \u0026lsquo;Moneymaker\u0026rsquo; plants was diminished by seed coating with bacterial antagonists (Figure 1), resulting in improved vegetative growth. This was achieved by improving the root\u0026apos;s ability to absorb water and nutrients under RKN stress (Farahat et al. 2012). As a result, the coated seed plants showed increased fresh weights of roots and shoots, along with greater stem length and diameter (Table 2). Additionally, the number of plant leaves (Table 2) and their photosynthetic pigment content (Table 3) increased, resulting in a larger green surface area for photosynthesis. Therefore, the fresh and dry weights of the plant shoots and roots increased (Table 2). Furthermore, some bacterial isolates may also have a stimulating effect on vegetative growth genes. Ayaz et al. (2021) found that the genesSlCKX1, SlIAA1, and Exp18 that promote plant growth were found to increase in expression in tomato plants treated with \u003cem\u003eB. atrophaeus\u003c/em\u003e GBSC56.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3.4. Plant metabolism activity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe plant metabolic activity, i.e., root and shoot content of dry matter and root content of carbohydrates, proteins, and polyphenols, of tomato \u0026lsquo;Moneymaker\u0026rsquo; coated and uncoated seed plants at 45 DAI presented in Table 4. The seed coating with several bacterial antagonists had a highly significant effect (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.001) on the metabolic activity of the \u0026lsquo;Moneymaker\u0026rsquo; plant under RKN infestation conditions in both seasons (Table 4). However, the CV values for the plant metabolic activity estimates were generally low, except for the root content of Pr in both seasons (23.29 and 14.98%, respectively), as well as SDM and RDM in 2019, which showed very high values (45.73 and 44.06%, respectively; Table 4).\u003c/p\u003e\n\u003cp\u003eSDM (mg g\u003csup\u003e-1\u003c/sup\u003e) ranged from 12.17 with coated seed plants by B1 and B2 to 13.04 with B14-coated seed plants in 2019, and from 10.49 with B7-coated seed plants to 15.98 with B18-coated seed plants in 2020 (Table 4). In both seasons, the lowest SDM was observed with coated seed plants by B1 (12.17 \u0026amp; 10.90, respectively) and B7 (12.52 \u0026amp; 10.49, respectively), with significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) among them and uncoated seed plants and coated seed plants by B2, B21, B9, and B10 in 2019 (12.38, 12.17, 12.45, 12.39, and 12.45, respectively), and those by B22 in 2020 (11.46). The highest SDM in both seasons was found with coated seed plants by B5 (12.72 \u0026amp; 15.44, respectively) and B19 (13.03 \u0026amp; 15.98, respectively), with significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) among them and coated seed plants by B1, B4, B6, B8, B12, B14, B16, B17, B22, B24, and B25 in 2020 only (ranging from 12.64 \u0026ndash; 13.04).\u003c/p\u003e\n\u003cp\u003eRDM (mg g\u003csup\u003e-1\u003c/sup\u003e) ranged from 21.03 with B14-coated seed plants to 22.60 with B7-coated seed plants in 2019, and from 18.57 with B4-coated seed plants to 30.94 with B18-coated seed plants in 2020. The highest RDM in both seasons was found with B8-coated seed plants (22.54 \u0026amp; 26.40, respectively), with significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) with uncoated seed plants and coated seed plants by B2, B7, and B21 in 2019 (22.20, 21.98, 22.60, and 22.43, respectively), and those by B9 and B19 in 2020 (29.03 \u0026amp; 30.94, respectively). In both seasons, the lowest RDM was found with coated seed plants B1, B3-B6, B10\u0026ndash;B13, B14-B16, B18, B22, B23, B24, and B25 with significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) among them and coated plants by B2, B9, B17, and B19 in 2019 (21.98, 21.56, 21.54, and 21.54, respectively), and uncoated and coated seed plants by B21 and B7 in 2020 (22.43, 23.53, and 18.70, respectively) (Table 4).\u003c/p\u003e\n\u003cp\u003eResults of the root content of carbohydrates, proteins, and polyphenols in tomato \u0026lsquo;Moneymaker\u0026rsquo; coated and uncoated seed plants are presented in Table 4. Coating the seed \u0026lsquo;Moneymaker\u0026rsquo; with several bacterial antagonists had a highly significant effect (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.001) on the root content of Carb, Pr, and PPh under RKN infestation conditions in both seasons (Table 4). The root Carb content (mg g\u003csup\u003e-1\u003c/sup\u003e) was between 41.56 with uncoated seed plants and 53.97 with B21-coated seed plants in 2019, and between 38.81 with uncoated seed plants and 50.31 with B19-coated seed plants in 2020 (Table 4). The root Carb content was inconsistent in both seasons, particularly with the highest content. In 2019, the highest root Carb content was found in B21-coated seed plants (53.97), followed by coated seed plants by B6 (53.59) and B13 (53.58), which have significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) among them, and then coated seed plants by B5, B9, and B19 (53.28, 53.25, and 53.25, respectively), which have significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) among them (Table 4). B19-coated seed plants had the highest root Carb content in 2020 (50.31), followed by B4-coated seed plants (50.17), and then B20-coated seed plants (50.05). The uncoated seed plants had the lowest root Carb content in both seasons (41.56 \u0026amp; 38.81, respectively), with significant similarities with coated seed plants by B1, B2, B8, and B10 in 2019 only (51.57, 51.73, 51.76, and 51.08, respectively), and coated seed plants by B5, B15, B18, and B23 in 2020 (48.40, 48.37, 47.85, and 48.29, respectively).\u003c/p\u003e\n\u003cp\u003eThe root Pr content (mg g\u003csup\u003e-1\u003c/sup\u003e) ranged from 8.81 with B5-coated seed plants to 19.10 with B18-coated seed plants in 2019, and from 10.07 with uncoated seed plants to 17.82 with B18-coated seed plants (Table 4). The highest root Pr content was found in B18-coated seed plants in both seasons (19.10 \u0026amp; 17.82, respectively), with significant similarities with coated seed plants by B10 and B15 in 2019 (19.09 \u0026amp; 19.08, respectively), and with B11-coated seed plants in 2020 (17.81). The lowest root Pr content was inconsistent in both seasons. In 2019, B5-coated seed plants and uncoated seed plants had the lowest content (8.81 and 10.80, respectively), with significant differences (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) among them. In 2020, the uncoated seed plants had the lowest content (10.07), followed by B19-coated plants (15.69).\u003c/p\u003e\n\u003cp\u003eThe root PPh content (mg g\u003csup\u003e-1\u003c/sup\u003e) ranged from 5.16 with uncoated seed plants to 6.74 with B21-coated seed plants in 2019, and from 4.82 with uncoated seed plants to 6.30 with B2-coated seed plants in 2020 (Table 4). The root PPh content was inconsistent in both seasons, particularly the highest content. The highest root PPh content was found in coated seed plants by B21 and B13 in 2019 (6.74 \u0026amp; 6.73, respectively), with significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) among them, and coated seed plants by B2, B17, and B19 in 2020 (6.30, 6.30, and 6.29, respectively) with significant similarities (\u003cem\u003eP\u003c/em\u003e \u0026lt;0.05) among them. The uncoated seed plants had the lowest root PPh content in both seasons (5.16 \u0026amp; 4.82, respectively).\u003c/p\u003e\n\u003cp\u003eRKN infestation in uncoated seed tomato \u0026lsquo;Moneymaker\u0026rsquo; plants resulted in a decrease in foliar photosynthetic pigments (Table 3), causing a decrease in photosynthesis and the production of carbohydrates and proteins (Khan et al. 2006, Campos et al. 2012). RKN also increased reactive oxygen species (ROS; Teresa Melillo et al. 2006), which elevated chlorophyllase activity, causing chlorophyll degradation and impairing cellular structures such as proteins, carbohydrates, lipids, and DNA (Fujimoto et al. 2021, Holbein et al. 2016). Therefore, leaves of uncoated seed \u0026lsquo;Moneymaker\u0026rsquo; plants have a higher chlor-a/chlor-b ratio in the leaves, likely due to chlorophyll-b being degraded to chlorophyll-a (Fang et al. 1998). RKN infestation also reduced dry matter content in both roots and shoots and lowered the carbohydrate and protein content in the roots, consistent with previous studies (Khan et al. 2022, Banora and Almaghrabi 2019). RKN-induced secretion of hydrolyzing enzymes may contribute to the decreased carbohydrate content in the roots (Anwar 1995, Farahat et al. 2012), while increased root protein content may be related to resistance mechanisms in the plants (Mukherjee et al. 2020, Patel and Patel 1995).\u003c/p\u003e\n\u003cp\u003ePolyphenols in resistant tomato plants are essential in defending against RKN by acting as signaling molecules to detect and neutralize ROS (Khan et al. 2022, Patel et al. 2017, Campos et al. 2012, Farahat et al. 2012) and by processing\u0026nbsp;nematicidal properties that reduce RKN damage (Pratysha 2022, Aissani et al. 2018). In this study, coated-seed tomato \u0026lsquo;Moneymaker\u0026rsquo; plants exhibited higher polyphenol contents in their roots compared to those with uncoated seeds. As a result, the coated-seed plants showed lower NG and NEM, as well as a higher content of foliar photosynthetic pigments, dry matter content in both shoots and roots, and root carbohydrates and proteins. Their ability to scavenge ROS may be the reason for this.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3.8. Multivariate analyses\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMultivariate analyses, particularly principal component analysis (PCA) and cluster analysis (CA), are often employed to evaluate variability among treatments, identify the correlations between traits and treatments, and categorize treatments based on estimated traits to select the best ones (Johnson and Wichern 2007). PCA is a valuable tool for dividing multiple traits into fewer components that explain the majority of the variance. In this study,\u0026nbsp;PCA was performed on 18 traits related to RKN-satisfying response, vegetative growth, and metabolic activity of uncoated and coated seed \u0026lsquo;Moneymaker\u0026rsquo; plants by several bacterial antagonists grown in a naturally RKN-infested greenhouse during the 2019 and 2020 fall seasons. Eighteen principal components (PCs) were identified with eigenvalues ranging from 5.65 to 0.00009, as shown in Figure 2. The first 6 PCs had eigenvalues \u0026gt;1 and contributed 94.08% of cumulative variability (Figure 2). All estimated traits had a significant impact on the first 6 PCs (Table 5), except PC5, suggesting that these traits of \u0026lsquo;Moneymaker\u0026rsquo; plants can be used to classify bacterial isolates (Johnson and Wichern 2007). PC1 (eigenvalue= 5.472) explained 30.40% of the variance (Figure 2) and was influenced negatively by traits SL (-0.225), SD (-0.240), NPL (-0.258), SFW (-0.225), SDW (-0.268), Chlor-b (-0.207), Carb (-0.331), Pr (-0.276), and PPh (-0.280), and being positively influenced by NG (0.310) and NEM (0.343) (Table 5). PC2 (eigenvalue = 2.395) explained 13.31% of the variance (Figure 2) and was positively correlated only with T-Carot (0.495) (Table 5). PC3 (eigenvalue = 2.228) was responsible for 12.74% of the variance (Figure 2) and had positive correlations with Chlor-a (0.460) and T-Chlor (0.505), but negative correlations with only RFW (-0.374) (Table 5). PC4 (eigenvalue = 1.934) was responsible for 10.743% of the variance (Figure 2) and had positive correlations with RDW (0.516) and RDM (0.435) (Table 5). PC5 (eigenvalue = 1.496) accounted for 8.31% of the variance (Figure 2) but was not influenced by any traits (Table 5). PC6 (eigenvalue = 1.098) explained 6.098% of the variance (Figure 2) and had a positive correlation with only SDM (0.508) (Table 5).\u003c/p\u003e\n\u003cp\u003eFigure 3 displays the biplot of the first two PCs, which depicts the distribution of coated and uncoated seed \u0026lsquo;Moneymaker\u0026rsquo; plants with bacterial isolates and the estimated traits under RKN infestation during the 2019 and 2020 fall seasons. The vectors for NG and NEM were grouped in a separate quadrant with similar directions, and angles between them less than 90\u0026deg;. The vectors for the residual traits were located in two quadrants that were opposite to each other, with the vectors close together and angled similarly (Figure 3). Thus, Pearson\u0026rsquo;s correlation coefficients were estimated. Significant correlations were discovered in Pearson\u0026rsquo;s correlation coefficients for estimated traits, as shown in Table 6. This study focused on the correlations between RKN-satisfying response, i.e., NG \u0026amp; NEM, and traits that related to vegetative growth and plant metabolic activity in coated and uncoated seed plants. Estimates of NG \u0026amp; NEM showed highly positive correlations among them. Also, NG \u0026amp; NEM showed negative correlations with SL (-0.429\u003csup\u003e*\u003c/sup\u003e\u0026amp;-0.511\u003csup\u003e*\u003c/sup\u003e, respectively), SFW (0.430\u003csup\u003e*\u003c/sup\u003e \u0026amp; 0.408\u003csup\u003e*\u003c/sup\u003e, respectively), SDW (0.437\u003csup\u003e*\u003c/sup\u003e \u0026amp; 0.421\u003csup\u003e*\u003c/sup\u003e, respectively), chlor-b (0.484\u003csup\u003e*\u003c/sup\u003e \u0026amp; 0.486\u003csup\u003e*\u003c/sup\u003e, respectively), Carb (0.395\u003csup\u003e*\u003c/sup\u003e \u0026amp; 0.501\u003csup\u003e*\u003c/sup\u003e, respectively), and Pr (0.440\u003csup\u003e*\u003c/sup\u003e \u0026amp; 0.529\u003csup\u003e*\u003c/sup\u003e, respectively) (Table 6). The photosynthetic pigment content in tomato leaves is reduced by RKN infestation, decreasing the photosynthesis rate and the accumulation of carbohydrates and proteins (Campos et al. 2012, Khan et al. 2006, 2022). Tomato plants are weakened by this, resulting in smaller leaves, shorter stems, and decreased yield components (Banora and Almaghrabi 2019). Previous studies have found negative correlations between tomato plant growth parameters and initial RKN population density (Mekete et al. 2003, Schomaker et al. 2006) and RKN reproduction factor (Gharabadiyan et al. 2013). Padilla-Hurtado et al. (2022) found a significantly negative correlation between the RKN-damage scale and the number of plant fruits. Sharma et al. (1990) and Patel et al. (2017) indicated that polyphenol synthesis in tomato plants is correlated with nematode infection.\u003c/p\u003e\n\u003cp\u003eThe coated and uncoated seed plants with several bacterial isolates were distributed across all quarters of the biplot (Figure 3), reflecting high variability among the bacterial isolates impact on RKN-satisfying response, vegetative growth, and metabolic activity of tomato \u0026lsquo;Moneymaker\u0026rsquo;Top of Form\u003c/p\u003e\n\u003cp\u003e. Treatments with higher trait values were positioned distant from the vector line, often at the vertices of the convenx hull (Johnson and Wichern 2007). Uncoated seed plants were plotted in a separate quadrant. The coated seed plants by B1, B2, B3, B5, B7, B8, B9, B10, B11, B22, and B25 were plotted in another quadrant. The direction of both groups was toward NG and NEM vectors. Coated seed plants by B6, B12, B21, B15, and B20 were plotted in the same quadrant close to the vector lines of SD, SL, T-Carot, SDM, RFW, RDW, RDM, Carb, PPh, and Pr. Coated seed plants by B4, B13, B14, B16, B17, B18, B19, and B24 were plotted in another quadrant close to the vector lines of NPL, Chlor-a, Chlor-b, T-Chlor, SFW, and SDW. Therefore, hierarchical cluster analysis (HCA) can be performed to categorize bacterial isolates based on their impacts on RKN-satisfying response, vegetative growth, and metabolic activity of tomato \u0026lsquo;Moneymaker\u0026rsquo; plants (Johnson and Wichern 2007).\u003c/p\u003e\n\u003cp\u003eHCA revealed five distinct clusters as shown in Figure 4. Cluster-1 included only coated seed plants by B25 (Figure 4), and was characterized by very low values of SL and NPL; low values of SFW, SDW, SDM, FRW, RDW, and RDM; moderate values of Chlor-a, Chlor-b, T-Chlor, and SDM; high NG and NEM; and very high values of SD, T-Carot, Carb, Pr, and PPh (Table 7). Cluster-2 consisted of coated seed plants by B1, B3, B5, B6, B7, B8, B9, B10, B11, B12, and B21 (Figure 4), which had low values of Chlor-b and SD, moderate values of NG, NEM, SL, SFW, SDW, RFW, RDW, Carb, Pr, and PPh; and high values of NPL, Chlor-a, T-Chlor, T-Carot, and SDM (Table 7). Cluster 3 consisted of coated seed plants by B2, B4, B13, B14, B15, B16, B17, B18, B19, B20, B22, and B24 (Figure 4), and was characterized by low values of NG and NEM; moderate values of SD, NPL, Chlor-a, Chlor-b, T-Chlor, T-Carot, and RDM; high values of SL, SFW, SDW, SDM, Carb, Pr, and PPh; and very high values of RFW and RDW (Table 7). Cluster 4 consisted of only coated seed plants by B23 (Figure 4), which had very low values of NG, NEM, and RDM; low values of T-Carot, Carb, and PPh; high values of SD, RFW, RDW, and Pr; and very high values of SL, NPL, Chlor-a, Chlor-b, T-chlor, SFW, SDW, and SDM (Table 7). Cluster 5 consisted of only uncoated plants (Figure 4), and was characterized by very high values of NG, NEM, and RDM, and vary low values of the rest traits (Table 7). Cluster centroids had different levels of separation, with the smallest between Clusters 2 and 3 (23.862) and the largest between Clusters 4 and 5 (82.553) (Table 7). The difference in trait profiles between clusters was emphasized by the variance, which ranged from 0 for Clusters 1, 4, and 5 to 198.116 for Cluster-3 (Table 7).\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe bacterial isolates\u003cem\u003eBacillus subtilis\u003c/em\u003e subsp. \u003cem\u003espizizenii\u003c/em\u003e Sd1-14, \u003cem\u003eStreptomyces subrutilus\u003c/em\u003e Wb2n-11, \u003cem\u003eStreptomyces scabiei\u003c/em\u003e WB1n-4, and \u003cem\u003eBacillus mojavensis\u003c/em\u003e Sd2Re-10 demonstrated strong effectiveness in both suppressing RKN and enhancing vegetative growth in tomato \u0026lsquo;Moneymaker\u0026rsquo; plants, as did well-known microbial biocontrol agents \u003cem\u003ePseudomonas trivialis\u003c/em\u003e 3Re2-7 and \u003cem\u003eSporosarcina psychrophila\u003c/em\u003e Sd4-11. Future research will evaluate the effects of these bacterial isolates, individually or in combination, on the growth of various tomato cultivars and other vegetable crops. The focus of the research will be on their potential for manage nematodes and soil-borne pathogens, while also promoting plant growth and maintaining soil microbal communities. Future studies will also aim to assess the extent and duration of soil colonization by these bacterial isolates to identify stable commercial formulations of them in line with national regulations.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contribution\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAMAM is responsible for conceptualization, formal analysis, software, supervision, validation, visualization, and writing of an original draft, review, and editing. AASAE is responsible for the conceptualization and supervision. MA is responsible for the conceptualization and supervision. MAAM is responsible for data curation, investigation, methodology, and writing an original draft.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAbd-Algawad MMM (2014) Plant-parasitic nematode threats to global food security. J Nemat 46:130.\u003c/li\u003e\n\u003cli\u003eAdam M, Westphal A, Hallmann J, Heuer H (2014) Specific microbial attachment to root-knot nematodes in suppressive soil. Appl Environm Microbiol 80:2679-2686.\u003c/li\u003e\n\u003cli\u003eAissani N, Balti R, Sebai H (2018) Potent nematicidal activity of phenolic derivatives on \u003cem\u003eMeloidogyne incognita\u003c/em\u003e. J Helmin 92:668-673. https://doi.org.10.1017/S0022149X17000918.\u003c/li\u003e\n\u003cli\u003eAli NI, Siddiqui IA, Shaukat SS, Zaki MJ (2002) Nematicidal activity of some strains of \u003cem\u003ePseudomonas\u003c/em\u003e spp. Soil Biol Biochem 34:1051-1058. https://doi.org/10.1016/S0038-0717(02)00029-9.\u003c/li\u003e\n\u003cli\u003eAmorim DJ, Tsujimoto TF, Baldo FB, Leite LG, Harakava R, Wilcken SRS, Gabia AA, Amorim DJ (2024) \u003cem\u003eBacillus\u003c/em\u003e, \u003cem\u003ePseudomonas\u003c/em\u003e and \u003cem\u003eSerratia\u003c/em\u003e control \u003cem\u003eMeloidogyne incognita\u003c/em\u003e (Rhabditida: \u003cem\u003eMeloidogynidae\u003c/em\u003e) and promote the growth of tomato plants. Rhizoshpere 31:100935. https://doi.org/10.1016/j.rhisph.2024.100935\u003c/li\u003e\n\u003cli\u003eAnwar SA (1995) Influence of \u003cem\u003eMeloidogyne incognita\u003c/em\u003e, \u003cem\u003eParatrichodours minor\u003c/em\u003e, and \u003cem\u003ePratylenchus scribneri\u003c/em\u003e on root-shoot growth and carbohydrates partitioning in tomato. Pakistan J Zool 27:105-113.\u003c/li\u003e\n\u003cli\u003eAyaz M, Zhao JT, Zhao W, Chi YK, Ali Q, Ali F, Khan AR, Yu Q, Yu JWW, Wu WC, Qi RD, Huang WK (2021) Biocontrol of plant parasitic nematodes by bacteria and fungi: A multi-omics approach for the exploration of novel nematicides in sustainable agriculture. Front Microbiol 15:1433716. https://doi.org/10.3389/fmicb.2024.1433716.\u003c/li\u003e\n\u003cli\u003eBanora MY, Almaghrabi OAA (2019) Differential response of some nematode-resistant and susceptible tomato genotypes to \u003cem\u003eMeloidogyne javanica\u003c/em\u003e infection. J Plant Protec Res 59:113-123. https://doi.org/10.24425/jppr.2019.126040.\u003c/li\u003e\n\u003cli\u003eBanora MY (2023) I,pacting of root-knot nematodes on tomato: Current status and potential horizons for its managing. In: Lops F (ed), Tomato Cultivation and Consumption \u0026ndash; Innovation and Sustainability. IntechOpen Limited, London, UK, pp.1-22. https://doi.org/10.5772/intechopen.112868.\u003c/li\u003e\n\u003cli\u003eBeah A, Kamara AY, Jibrin JM, Akinseye FM, Tofa AI, Adam AM (2021) Simulating the response of drought-tolerant maize varieties to nitrogen application in contrasting environments in the Nigeria Savannas using the APSIM model. Agronomy 11(1):76. https://doi.org/10.3390/agronomy11010076.\u003c/li\u003e\n\u003cli\u003eCampos VAC, Machado ART, Oliveira DF, Campos VP, Chages RCR, Nunes AS (2012) Change in metabolites in plant roots after inoculation with \u003cem\u003eMeloidogyne incognita\u003c/em\u003e. Nematology 14:579-588. https://doi.org/10.1163/15684111x614494.\u003c/li\u003e\n\u003cli\u003eCastellano-Hinojosa A, Noling JW, Bui HX, Desaeger JA, Strauss SL (2022) Effect of fumigants and non-fumigants on nematode and weed control, crop yield, and soil microbial diversity and predicted functionality in a strawberry production system. Science of the Total Environment 852:158285. https://doi.org/10.1016/j.scitotenv.2022.158285.\u003c/li\u003e\n\u003cli\u003eCharchar JM, Gonzaga V, Giordano LB, Boiteux LS, dos Reis NV, de Arag\u0026atilde;o, FA (2003) Reaction of tomato cultivars to infection by a mixed population of \u003cem\u003eM. incognita\u003c/em\u003e race 1 and \u003cem\u003eM. javanica\u003c/em\u003e in the field. Nemat Bras 27:49-54.\u003c/li\u003e\n\u003cli\u003eCheng W, Yang J, Nie Q, Huang D, Yu C, Zheng L, Cai M, Thomashow L, Weller DM, Yu Z, Zhang J (2017) Volatile organic compounds from \u003cem\u003ePaenibacillus polymyxa\u003c/em\u003e KM2501-1 control \u003cem\u003eMeloidogyne incognita\u003c/em\u003e by multiple strategies. Sci Rep 24:16213. https://doi.org/10.1038/s41598-017-16631-8.\u003c/li\u003e\n\u003cli\u003eColagiero M, Rosso LC, Ciancio A (2018) Diversity and biocontrol potential of bacterial consortia associated to root-knot nematodes. Biol. Control 120:11\u0026ndash;16. https://doi.org/10.1016/j.biocontrol.2017.07.010.\u003c/li\u003e\n\u003cli\u003ed\u0026rsquo;Errico G, Marra R, Crescenzi A, Davino SW, Fanigliulo A, Woo SL, Lorito M 2019) Integrated management strategies of \u003cem\u003eMeloidogyne incognita\u003c/em\u003e and \u003cem\u003ePseudopyrenochaeta lycopersici\u003c/em\u003e on tomato using a \u003cem\u003eBacillus firmus\u003c/em\u003e-based product and two synthetic nematicides in two consecutive crop cycles in greenhouse. Crop Protect 122:159\u0026ndash;164. https://doi.org/10.1016/j.cropro.2019.05.004.\u003c/li\u003e\n\u003cli\u003eFang Z, Bouwkamp J, Solomos T (1998) Chlorophyllase activities and chlorophyll degradation during leaf senescence in non-yellowing mutant and wild type of \u003cem\u003ePhaseolus vulgaris \u003c/em\u003eL. J Exp Bot 49:503\u0026ndash;10. https://doi.org/10.1093/jxb/49.320.503.\u003c/li\u003e\n\u003cli\u003eFarahat AA, Alsayed AA, El-Beltagi HS, Mahfoud NM (2012) Impact of organic and inorganic fertilizers on nematode reproduction and biochemical alternations on tomato. Not Sci Biol 4:48-55.\u003c/li\u003e\n\u003cli\u003eFujimoto T, Abe H, Mizukubo T, Seo S. 2021. Phytol, a constituent of chlorophyll, induces root-knot nematode resistance in Arabidopsis via the ethylene signaling pathway. Mol Plant Mic Inter 34:279-285. https://doi.org/10.1094/MPMI-07-20-0186-R.\u003c/li\u003e\n\u003cli\u003eGhahremani Z, Escudero N, Beltr\u0026aacute;n-Anad\u0026oacute;n D, Saus E, Cunquero M, Andilla J, Loza-Alvarez P, Gabald\u0026oacute;n T, Sorribas FJ (2020) \u003cem\u003eBacillus firmus\u003c/em\u003e strain I-1582, a nematode antagonist by itself and through the plant.Front Plant Sci 11:796. https://doi.org/10.3389/fpls.2020.00796.\u003c/li\u003e\n\u003cli\u003eGharabadiyan F, Jamali S, Komeili HR (2013) Determination of rook-knot nematode (\u003cem\u003eMeloidogyne javanica\u003c/em\u003e) damage function for tomato cultivars. J Agr Sci. 58:147-157. https://doi.org/10.2298/JAS1302147G.\u003c/li\u003e\n\u003cli\u003eHashem M, Abo-Elyousr K (2011) Management of the root-knot nematode \u003cem\u003eMeloidogyne incognita\u003c/em\u003e on tomato with combinations of different biocontrol organisms. Crop Protect 30:285-292. https://doi.org/10.1016/j.cropro.2010.12.009.\u003c/li\u003e\n\u003cli\u003eHooper DU, Solan M, Symstad A, Diaz S, Gessner MO, Buchmann N, Degrange V, Grime P, Hulot F, Mermillod-Blondin F, Roy J, Sephn E, van Peer L (2005) Species diversity, functional diversity, and ecosystem functioning. In: Loreau M, Naeem S, Lnchausti P (Eds), Biodiversity and Ecosystem Functioning: Synthesis and Perspectives, Oxford University Press, pp. 195-281.\u003c/li\u003e\n\u003cli\u003eHu Y, You J, Wang Y, Long Y, Wang S, Pan F, Yu Z (2022) Biocontrol efficacy of \u003cem\u003eBacillus velezensis\u003c/em\u003e strain YS-AT-DS1 against the root-knot nematode \u003cem\u003eMeloidogyne incognita\u003c/em\u003e in tomato plants. Front Microbiol 13:1035748. https://doi.org/10.3389/fmicb.2022.1035748.\u003c/li\u003e\n\u003cli\u003eHussey RS, Barker KR (1973) A comparison of methods of collecting inocula of \u003cem\u003eMeliodogyne \u003c/em\u003espp., including a new technique. Plant Dis Rep 57:1025-1028.\u003c/li\u003e\n\u003cli\u003eJamal Q, Cho JY, Moon JH, Munir S, Anees M, Kim KY (2017) Identification for the first time of cyclo (D-Pro-L-Leu) produced by \u003cem\u003eBacillus amyloliquefaciens\u003c/em\u003e y1 as a nematocide for control of \u003cem\u003eMeloidogyne incognita\u003c/em\u003e. Molecules 22:1-16. https://doi.org/10.3390/molecules22111839.\u003c/li\u003e\n\u003cli\u003eJim\u0026eacute;nez-Aguirre MA, Padilla-Hurtado BE, Ceballos-Aguirre N, Cardona-Agudelo LD, Montoya-Estada CN (2023) Antagonist effect of native bacteria of the genus \u003cem\u003eBacillus\u003c/em\u003e on the root-knot nematode (\u003cem\u003eMeliodogyne \u003c/em\u003espp.) in tomato germplasm. Chil J Agric Res 83:565-576. http://dx.doi.org/10.4067/s0718-58392023000500565.\u003c/li\u003e\n\u003cli\u003eJin N, Xue H, Li W, Wang X, Liu Q, Liu S, Liu P, Zhao J, Jian H (2017) Field evaluation of \u003cem\u003eStreptomyces rubrogriseus\u003c/em\u003e HDZ-9-47 for biocontrol of \u003cem\u003eMeloidogyne incognita\u003c/em\u003e on tomato. J Integ Agric 16:1347\u0026ndash;1357. https://doi.org/10.1016/S2095-3119(16)61553-8.\u003c/li\u003e\n\u003cli\u003eJohnson RA, Wichern DW (2007) Applied multivariate statistical analysis, 794. 6\u003csup\u003end\u003c/sup\u003e ed. Englewood Cliffs, NJ: Prentice Hall.\u003c/li\u003e\n\u003cli\u003eKhan MR, Khan SM, Mohiddin FA, Askary TH (2006) Effects of high nickel soil on root-knot nematode disease of tomato. Nematropica 36:79-87.\u003c/li\u003e\n\u003cli\u003eKhan MR, Mohiddin FA, Ejaz NA, Khan MM (2012) Management of root-knot disease in eggplant through the application of biocontrol bacteria and neem leaves. Turk J Biol 36:161-169.\u003c/li\u003e\n\u003cli\u003eKhan MR, Sharma DN, Ahmad I (2022) Temporal impact of root-knot nematode infection on some important biochemical and physiological characters of tomato. Ind Phytopathol 75:749-758. https://doi.org/10.1007/s42360-022-00500-0.\u003c/li\u003e\n\u003cli\u003eK\u0026ouml;berl M (2010) Analyse von Arzneipflanzen-assoziierten Mikroorganismen: Biodiversit\u0026auml;t und antagonistisches Potenzial gegen bodenb\u0026uuml;rtige Phytopathogene. MSc, an der Naturwissenschaftlichen Fakult\u0026auml;t der Karl-Franzens-Universit\u0026auml;t Graz, 92p (In Deutsch).\u003c/li\u003e\n\u003cli\u003eK\u0026ouml;berl M, M\u0026uuml;ller M, Ramadan EM, Berg G (2011) Desert farming benefits from microbial potential in arid soils and promotes diversity and plant health. PloS One 6:e24452. https://doi.org/10.1371/journal.pone.0024452.\u003c/li\u003e\n\u003cli\u003eK\u0026ouml;berl M, Ramadan EM, Adam M, Cardinale M, Hallmann J, Heuer H, Smalla K, Berg G (2013) \u003cem\u003eBacillus\u003c/em\u003e and \u003cem\u003eStreptomyces\u003c/em\u003e were selected as broad-spectrum antagonists against soilborne pathogens from arid areas in Egypt. FEMS Microbiol 342:168-178. https://doi.org/10.1111/1564-6968.12089.\u003c/li\u003e\n\u003cli\u003eLatimer GW (2012) Official Methods of Analysis of AOAC International. 19\u003csup\u003eth\u003c/sup\u003e ed, The Association of Official Analysis Chemists International, Maryland, USA. p1399.\u003c/li\u003e\n\u003cli\u003eLee YS, Kim KY (2016) Antagonistic potential of \u003cem\u003eBacillus pumilus\u003c/em\u003e L1 against root-knot nematode, \u003cem\u003eMeloidogyne arenaria\u003c/em\u003e. J Phytopathol 164:29\u0026ndash;39. https://doi.org/10.1111/jph.12421\u003c/li\u003e\n\u003cli\u003eLi X, Hu HJ, Li JY, Wang C, Chen SL, Yan SZ (2019) Effects of the endophytic bacteria \u003cem\u003eBacillus cereus\u003c/em\u003e BCM2 on tomato root exudates and \u003cem\u003eMeloidogyne incognita\u003c/em\u003e infection. Plant Dis 103:1551\u0026ndash;1558. https://doi.org/10.1094/PDIS-11-18-2016-RE.\u003c/li\u003e\n\u003cli\u003eLopes-Caitar VS, Pinheiro JB, Marcelino-Guimar\u0026atilde;es FC (2019) Nematodes in horticulture: An overview. J Hort Sci Crop Res. 1:106.\u003c/li\u003e\n\u003cli\u003eMartinuz A, Schouten A, Sikora RA (2012) Systemically induced resistance and microbial competitive exclusion: Implications on biological control. Phytopathology 102:260- 266. https://doi.org/10.1094/PHYTO-04-11-0120.\u003c/li\u003e\n\u003cli\u003eMekete T, Mandefro W, Greco N (2003) Relationship between initial population densities of \u003cem\u003eMeloidogyne javanica\u003c/em\u003e and damage to pepper and tomato in Ethiopia. Nematol Mediter 31:169‒171.\u003c/li\u003e\n\u003cli\u003eMoran R (1982) Formulae for determination of chlorophyllous pigments extracted with \u003cem\u003eN\u003c/em\u003e,\u003cem\u003eN\u003c/em\u003e-dimethylformamide. Plant Phys 69:1376\u0026ndash;1381. https://doi.org/10.1104/pp.69.6.1376.\u003c/li\u003e\n\u003cli\u003eMukherjee A, Mondal P, Sinha Babu SP (2020) Nematode extract-induced resistance in tomato against \u003cem\u003eMeloidogyne incognita\u003c/em\u003e. Indian J Sci Tech. 13:1476-1479. https://doi.org/10.17485/IJST/v13i14.234.\u003c/li\u003e\n\u003cli\u003ePadilla-Hurtado B, Morillo-Coronado Y, Tarapues S, Burbano S, Soto-Su\u0026aacute;rez M, Urrea R, Ceballos-Aguirre N (2022) Evaluation of root-knot nematodes (\u003cem\u003eMeliodogyne \u003c/em\u003espp.) population density for disease resistance screening of tomato germplasm carrying the gene \u003cem\u003eMi-1\u003c/em\u003e. Chil J Agric Res 82:157-166. https://dx.doi.org/10.4067/S0718-58392022000100157.\u003c/li\u003e\n\u003cli\u003ePandey SK, Singh H (2011) A simple, cost-effective method for leaf area estimation. J Bot 2011:1\u0026ndash;6. https://doi.org/10.1155/2011/658240.\u003c/li\u003e\n\u003cli\u003ePatel VS, Shukla YM, Dhruye JJ (2017) Influence of rrot-knot nematode (\u003cem\u003eMeliodogyne \u003c/em\u003espp.) on phenolic acid profile in root of tomato (\u003cem\u003eSolanum lycopersicum\u003c/em\u003e L.). Inter J Curr Microb Appl Sci 6:840-848. https://doi.org/10.20546/ijcmas.2017.610.100.\u003c/li\u003e\n\u003cli\u003eScherwinski K, Grosch R, Berg G (2008) Effect of bacterial antagonists on lettuce: active biocontrol of \u003cem\u003eRhizoctonia solani\u003c/em\u003e and negligible, short-term effects on nontarget microorganisms. FEMS Microbiol Ecol 64:106-116. https://doi.org/10.1111/j.1574-6941.2007.00421.x\u003c/li\u003e\n\u003cli\u003eSeinhorst W (1966) Killing nematodes for taxonomic study with hot F.A.A. Nematol 12:178.\u003c/li\u003e\n\u003cli\u003eSharma JL, Trevidi PC, Sharma MK, Jiagi B (1990) Alteration in prolin and phenol content of \u003cem\u003eMeloidogyne incognita\u003c/em\u003e infected bringal cultivars. Pakist JNematol 8:33\u0026ndash;38.\u003c/li\u003e\n\u003cli\u003eSharma M, Devi S, Chand S (2025) Biocontrol strategies for sustainable management of root-knot nematodes. Physiol Molec Plant Pathol 136:102548. https://doi.org/10.1016/j.pmpp.2024.102548.\u003c/li\u003e\n\u003cli\u003eSharma M, Jasrotia S, Ohri P, Manhas RK (2019) Nematicidal potential of \u003cem\u003eStreptomyces antibioticus\u003c/em\u003e strain M7 against \u003cem\u003eMeloidogyne incognita\u003c/em\u003e. AMB Express 9:168. https://doi.org/10.1186/s13568-019-0894-2.\u003c/li\u003e\n\u003cli\u003eSiddiqui ZA, Sharma B, Siddiqui S (2007) Evaluation of \u003cem\u003eBacillus\u003c/em\u003e and \u003cem\u003ePseudomonas\u003c/em\u003e isolated for the biocontrol of \u003cem\u003eMeloidogyne incognita\u003c/em\u003e on tomato. Acta Phytopahol Entomol Hungar 42:25-34. https://doi.org/10.1556/APhyto.42.2007.1.4.\u003c/li\u003e\n\u003cli\u003eStirling GR, Mankau R (1978) Parasitism of \u003cem\u003eMeliodogyne \u003c/em\u003eeggs by a new fungal parasite. J Nematol 10:236-240.\u003c/li\u003e\n\u003cli\u003eTaylon AA, Dropkin H, Marin GC (1956) Perineal patterns of the root-knot nematodes. Phytopathology 46:26-34.\u003c/li\u003e\n\u003cli\u003eTeresa Melillo M, Leonetti P, Bongiovanni M, Catagnone-Sereno P, Bleve-Zacheo T (2006) Modulation of reactive oxygen species activities and H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e accumulation during compatible and incompatible tomato root-knot nematode interactions. New Phytol 170:501-512. https://doi.org/10.1111/j.1469-8137-2006.01724.x.\u003c/li\u003e\n\u003cli\u003eViaene N, Coyne DL, Kerry BR (2006) Biological and cultural management. In: Perry RN, Moens M (eds), Plant Nematology, CABI, Wallingford, UK, pp. 346-369.\u003c/li\u003e\n\u003cli\u003eViljoen JJF, Labuschagne N, Fourie H, Sikora RA (2019) Biological\u003ccite\u003e control of the root-knot nematode \u003c/cite\u003e\u003cem\u003eMeloidogyne incognita\u003c/em\u003e\u003ccite\u003e on tomatoes and carrots by plant growth-promoting rhizobacteria. \u003c/cite\u003e\u003cem\u003eTrop Plant Pathol\u003c/em\u003e\u003ccite\u003e 44:284\u0026ndash;291. \u003c/cite\u003ehttps://doi.org/10.1007/s40858-019-00283-2.\u003c/li\u003e\n\u003cli\u003eWickens TD, Keppel G (2004) Design and Analysis: A Researcher\u0026rsquo;s Handbook. Pearson Prentice-Hall, New Jersey, USA. 624p.\u003c/li\u003e\n\u003cli\u003eXia Y, Li S, Zhang C, Xu J, Chen Y (2019) B\u003cem\u003eacillus halotolerans \u003c/em\u003estrain LYSX1-induced systemic resistance against the root-knot nematode \u003cem\u003eMeloidogyne javanica\u003c/em\u003e in tomato. Ann Microbiol 69:1227-1233. https://doi.org/10.1007/s13213-019-01504-4.\u003c/li\u003e\n\u003cli\u003eXiao L, Wan JW, Yao JH, Feng H, Wei LH (2018) Effects of \u003cem\u003eBacillus cereus\u003c/em\u003e strain Jdm1 on \u003cem\u003eMeloidogyne incognita\u003c/em\u003e and the bacterial community in tomato rhizosphere soil. 3 Biotech 8:1\u0026ndash;8. https://doi.org/10.1007/s13205-018-1348-2.\u003c/li\u003e\n\u003cli\u003eYan W, Kang MS (2003) GCE-Biplot Analysis: A Graphical Tool for Breeders, Geneticists, and Agronomists. CRC Press, New York, USA.\u003c/li\u003e\n\u003cli\u003eYoon GY, Lee YS, Lee SY, Park RD, Hyun HN, Nam Y, Kim KY (2012) Effects on \u003cem\u003eMeloidogyne incognita\u003c/em\u003e of chitinase, glucanase and a secondary metabolite from \u003cem\u003eStreptomyces cacaoi\u003c/em\u003e GY525. Nematology 14:175-184. https://doi.org/10.1163/138855411X584124.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003eTables 1 to 7 are available in the Supplementary Files section.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"egyptian-journal-of-biological-pest-control","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"ebpc","sideBox":"Learn more about [Egyptian Journal of Biological Pest Control](http://ejbpc.springeropen.com)","snPcode":"41938","submissionUrl":"https://submission.springernature.com/new-submission/41938/3","title":"Egyptian Journal of Biological Pest Control","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Open","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Bacillus, Meloidogyne, Polyphenols, Paenibacillus, Streptomyces, Solanum lycopersicum","lastPublishedDoi":"10.21203/rs.3.rs-6043078/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6043078/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground:\u003c/strong\u003e Tomato yield is significantly reduced by root-knot nematodes (RKN; \u003cem\u003eMeloidogyne\u003c/em\u003e spp.), particularly in tropical and subtropical regions. This study evaluated 20 bacterial isolates (B1-B20), belonging to the genera \u003cem\u003eBacillus\u003c/em\u003e, \u003cem\u003eLysobacter\u003c/em\u003e, \u003cem\u003ePaenibacillus\u003c/em\u003e, and \u003cem\u003eStreptomyces\u003c/em\u003e, from Sekem farms in Egypt for their potential to biocontrol RKN and stimulate plant growth in tomato ‘Moneymaker’. The bacteria were compared with well-known microbial biocontrol agents (MBA), including \u003cem\u003eRhizobium etli\u003c/em\u003e G12 (B21), \u003cem\u003ePseudomonas trivialis\u003c/em\u003e 3Re2-7 (B22), \u003cem\u003eSporosarcina psychrophile\u003c/em\u003e Sd4-11 (B23), and \u003cem\u003eB. subtilis\u003c/em\u003e Sb1-20 (B24), and a negative control \u003cem\u003eEscherichia coli\u003c/em\u003e JM109 (B25). The study involved seed-coated and uncoated plants with bacterial isolates, planted in plastic pots, and inoculated with 1500 \u003cem\u003eM. incognita\u003c/em\u003e J\u003csub\u003e2\u003c/sub\u003e individuals per pot. Plants were grown in a saran-house during the 2019 and 2020 fall seasons, and their RKN-satisfying response (number of galls: NG and egg masses: NEM), vegetative growth, and metabolic activity were assessed 45 days after inoculation.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e In both seasons, seed coating with bacterial isolates achieved a significant improvement in plant growth (coefficient of variation: CV ranging 26.8-120.2% in 2019 and 10.9-48.8% in 2020) and a reduction in RKN-satisfying response (CV for NG: 57.6 and 53.8%, respectively; and for NEM: 56.5 and 65.3%, respectively). Compared to uncoated-seed plants, the bacterial seed coating reduced NG by 0.66-74.09% in 2019 and 14.61-66.29% in 2020. Similarly, NEM decreased by 0.63-70.61% in 2019 and 41.91-77.46% in 2020. The coated-seed plants by \u003cem\u003eBacillus subtilis \u003c/em\u003esubsp. \u003cem\u003espizizenii\u003c/em\u003e (B5), \u003cem\u003eStreptomyces subrutilus \u003c/em\u003eWb2n-11 (B12), \u003cem\u003eStreptomyces scabiei\u003c/em\u003e (B19), and \u003cem\u003eBacillus mojavensis\u003c/em\u003e (B20), along with the well-known MBAs B22 and B23, showed increased photosynthetic pigments, fresh weight of roots and shoots, stem size, and number of leaves. This growth has also led to higher dry weights in roots and shoots, and an increase in the root content of carbohydrates and proteins. Seed coating induced systemic RKN resistance by increasing polyphenol in root. In contrast, uncoated-seed plants showed reduced foliar photosynthesis pigment and metabolic activity due to high RKN damage. Principal component analysis revealed significant correlations between the evaluated traits. Hierarchical clustering categorized bacteria isolates into five clusters based on their impact on estimated plant traits.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion: B5, B12, B19, B20, B22, and B23\u003c/strong\u003e demonstrated superior performance in both controlling RKN and stimulating vegetative growth in tomato ‘Moneymaker’ plants as known MBAs.\u003c/p\u003e","manuscriptTitle":"Biocontrol of Meloidogyne incognita and Vegetative Growth Stimulation in Tomato ‘Moneymaker’ Plants by Egyptian Soil Bacteria","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-04-17 05:27:53","doi":"10.21203/rs.3.rs-6043078/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-04-15T14:33:07+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-04-15T08:54:06+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"245319709117545098286401616323664428754","date":"2025-04-15T08:26:32+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-04-15T08:23:41+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-04-10T12:34:38+00:00","index":"","fulltext":""},{"type":"submitted","content":"Egyptian Journal of Biological Pest Control","date":"2025-04-09T21:36:42+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"egyptian-journal-of-biological-pest-control","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"ebpc","sideBox":"Learn more about [Egyptian Journal of Biological Pest Control](http://ejbpc.springeropen.com)","snPcode":"41938","submissionUrl":"https://submission.springernature.com/new-submission/41938/3","title":"Egyptian Journal of Biological Pest Control","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Open","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"f8148965-1f9c-43bd-be62-c30ff7c6eb4f","owner":[],"postedDate":"April 17th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-06-16T16:04:41+00:00","versionOfRecord":{"articleIdentity":"rs-6043078","link":"https://doi.org/10.1186/s41938-025-00860-5","journal":{"identity":"egyptian-journal-of-biological-pest-control","isVorOnly":false,"title":"Egyptian Journal of Biological Pest Control"},"publishedOn":"2025-06-13 15:57:51","publishedOnDateReadable":"June 13th, 2025"},"versionCreatedAt":"2025-04-17 05:27:53","video":"","vorDoi":"10.1186/s41938-025-00860-5","vorDoiUrl":"https://doi.org/10.1186/s41938-025-00860-5","workflowStages":[]},"version":"v1","identity":"rs-6043078","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6043078","identity":"rs-6043078","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}