Use of Microbial Fertilizers in Grapevine Seedling Production

Use of Microbial Fertilizers in Grapevine Seedling Production | 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 Use of Microbial Fertilizers in Grapevine Seedling Production İsa Hazar, adem yağcı, rüstem cangi, Selda Daler This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8475362/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract In Türkiye, grafted grapevine seedling production remains limited, and maintaining high seedling quality continues to be a major challenge. This study evaluated the effects of different microbial fertilizer (MF) formulations on seedling yield and quality in grafted grapevine production over two consecutive years (2024–2025). Scions of Vitis vinifera L. cv. Narince were grafted onto the 110 R rootstock and used as plant material. Seven microbial fertilizer treatments and two control groups (positive and negative) were tested. The microbial fertilizers contained either single microorganisms ( Methylobacterium symbioense, Glomus fasciculatum, Glomus iranicum, Bacillus megaterium, and Trichoderma harzianum ) or microbial consortia ( Pseudomonas fluorescens, Paenibacillus polymyxa, Pantoea agglomerans, and Glomus spp.). Treatments were applied to the basal ends of cuttings before planting. Seedling yield, shoot and root growht parameters were assessed two months after planting. Microbial fertilizers did not significantly affect shoot growth in the first year, whereas all treatments enhanced shoot growth in the second year compared with the controls. Root length, fresh root weight, and dry root weight increased significantly under all microbial fertilizer treatments. The highest total and first-grade seedling yields were obtained from the positive control (74.1% and 57.8%) and MF-B (71.2% and 57.0%) treatments. Strong positive correlations were observed between fresh and dry root weight (r = 0.96) and between first grade and total seedling yield (r = 0.96). Principal component analysis indicated that root morphology and seedling yield traits explained most of the total variation. These results demonstrate that microbial fertilizers can substantially improve seedling quality and production efficiency in grafted grapevine propagation by enhancing root system development. Vitis vinifera L. Grafting Seedling yield Microbial inoculation Root Shoot Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Introduction Grapevine ( Vitis vinifera L.) is one of the most economically and culturally important crops in Türkiye, where nursery production constitutes the structural foundation of vineyard establishment (Korkutal et al. 2009 ). Nevertheless, grapevine nursery production continues to face major challenges, including limited availability of certified, disease-free and true-to-type planting material, which directly affects production efficiency and cost (Bahar et al. 2006 ; Durmaz et al. 2019 ; Uysal and Ateş 2016 ). In addition, structural limitations within the Turkish nursery sector, such as production capacity and marketing conditions, further constrain producer competitiveness (Cangi et al. 2018 ). Consequently, the demand for innovative and sustainable biological inputs has increased. Among the main factors influencing seedling yield and quality, fertilization practices play a central role. Environmental concerns associated with conventional fertilization and its long-term negative impacts on soil health have accelerated the adoption of eco-friendly alternatives. In this context, microbial fertilizers and plant biostimulants have attracted considerable attention due to their potential to enhance plant growth and productivity. Biostimulants are generally classified as microbial and non-microbial, with microbial biostimulants primarily consisting of arbuscular mycorrhizal fungi (AMF), plant growth–promoting rhizobacteria (PGPR) and Trichoderma spp. (Van Loon 2007 ; Rouphael and Colla 2020 ; Tsvetkov et al. 2014 ). Plant growth is largely governed by root–soil interactions, in which roots and microorganisms engage in both nutrient competition and mutualistic symbiosis (Belal et al. 2023 ; Martin and Van der Heijden 2024 ). Most terrestrial plant species form associations with mycorrhizal fungi that enhance nutrient and water uptake in exchange for plant-derived carbon (Sandal Erzurumlu and Kara 2014; Agnolucci et al. 2019 ; Moukarzel et al. 2024 ). Microbial biostimulants have been extensively investigated in grapevine propagation. Mycorrhizal preparations have been evaluated in grapevine cuttings, rootstocks and grafted seedlings (Gendiah 1991; Kara and Özdemir 2009 ; Kavak 2006 ; Kara et al. 2011 ; Eftekhari et al. 2010 ; Eroğlu and Çelik 2013 lıç and Cangi 2019), as well as under vineyard conditions, with variable effects on growth and nutrient acquisition (Camprubí et al. 2008 ). Beneficial microorganisms commonly used in biofertilizers include Bacillus , Azotobacter , Trichoderma , Rhizobium , Azospirillum and Saccharomyces spp., among which Trichoderma spp. are particularly well studied due to their combined roles in biocontrol and plant growth promotion (Calvo et al. 2014 ; Di Marco and Osti 2008 ; Cangi et al. 2020 ). Numerous studies have demonstrated that Trichoderma spp. stimulate plant growth, improve root and shoot development, enhance yield, increase tolerance to abiotic stresses, promote nutrient use efficiency and enhance photosynthetic activity (Yedidia et al. 2001 ; Harman 2006 ; Woo et al. 2006 ; Di Marco and Osti 2007 ; Tsvetkov et al. 2014 ; Mahmood 2015 ; Korkutal et al. 2017 ). Similarly, PGPR applications on grapevine rootstocks have been shown to improve vegetative growth, mineral nutrition and chlorophyll content while reducing chemical fertilizer requirements (Sabir et al. 2012 ), thereby supporting sustainable production systems (Rouphael and Colla 2020 ). Plant growth–promoting microorganisms enhance plant performance through phytohormone production, improved mineral acquisition, biological nitrogen fixation and phosphorus solubilization, although their effectiveness depends strongly on microbial composition, application rate, plant genotype and environmental conditions (Lucy et al. 2004 ; Etesami and Maheshwari 2018 ; Lopes et al. 2021 ; Etesami and Glick 2024 ; Khan et al. 2024 ; Nagrale et al. 2023 ). Therefore, systematic evaluation of microbial fertilizer formulations in economically important crops such as grapevine, particularly at the nursery stage, remains essential. The objective of this study was to determine the effects of different microbial fertilizers on shoot and root development and overall seedling quality in container-grown grafted grapevine production. We hypothesized that the application of different microbial fertilizer formulations would significantly enhance root system development and sapling quality of grafted grapevine ( Vitis vinifera L. cv. Narince/110 R) compared with control groups (positive and negative controls), and that the magnitude of these effects would vary among microbial consortia and across years due to environmental modulation of root–microbe interactions. Materials and Methods Experimental Site and Plant Material The experiment was conducted over the 2024 and 2025 propagation seasons in the grapevine seedling facilities and greenhouse units of the Agricultural Practice and Research Center at Tokat Gaziosmanpaşa University (TOGU), Türkiye. The plant material consisted of the wine grape cultivar Narince grafted onto the 110 R rootstock (V. berlandieri Rességuier No.2 × V. rupestris Martin 110 Richter), a genotype widely recognized for its tendency to root poorly (Howell 1987 ). Rootstock cuttings were supplied by the Manisa Viticulture Research Institute, whereas scion cuttings were collected from the TOGU research vineyard. Rootstock cuttings were prepared from the well-matured mid-sections of one-year-old dormant shoots in accordance with the approach described by Roux (1988). Following TS-4027 criteria, 110 R cuttings were selected at 30–40 cm in length and 8–12 mm in diameter. The Narince scion shoots were roughly 100 cm long and averaged 15 buds. All cuttings were immersed in a fungicide solution (1 kg Switch 62.5 WG per m³ water), sealed in polyethylene bags, and maintained at + 4°C with 80–95% relative humidity for approximately 60 days until grafting (Ağaoğlu and Çelik 1978 ). Grafting Procedures and Callusing Conditions Five days before grafting, cuttings were removed from cold storage and allowed to acclimate gradually to room temperature. To compensate for moisture depletion during storage, rootstock and scion cuttings were soaked in fungicide-amended water for 24 and 48 hours, respectively (Sucu et al. 2018 ). Thermotherapy was then applied to reduce potential pathogen load: scions were treated for 15 minutes at 50°C, and rootstocks for 30 minutes (Sağlam 2017 ). Grafting was performed using a pedal-operated Ω-type grafting machine. Immediately afterward, the upper 5–7 cm of each grafted cutting, including the graft union, was coated with paraffin at 74–76°C for 1–2 seconds. This step helped prevent dehydration, reduced contamination risk, and supported the initial callus formation process (Richards 1976 ). To avoid any heat-related bud injury, each cutting was briefly cooled in water for 1–2 seconds (Küçükyumuk 2009). Fungicide-moistened pine sawdust (30 g Switch 62.5 WG in 15 L water) was used as the bedding medium during callusing. Grafted cuttings were placed in plastic trays layered with sawdust and transferred to a callusing room for a 21-day period. During this time, temperature and humidity gradually decreased: 27 ± 1°C and 80–85% RH for the first 3 days, 25 ± 1°C and 80–82% RH for the following 15 days, 23 ± 1°C and 75–80% RH for the final 3 days (Aslan et al., 2015 ). Following callusing, the cuttings were slowly acclimatized for about one week. Sawdust was removed using pressurized air, and a second paraffin dip (80–82°C) was applied to maintain callus moisture. Prior to planting, the basal 10 cm of each cutting was soaked in water for 3 days. Microbial Fertilizer Treatments and Planting The experimental design included seven microbial fertilizer treatments along with a positive control (Indole-3-butyric acid) and an untreated negative control. The microbial compositions of the fertilizers used in the study are presented in Table 1 . After hydration and preparation steps were completed, the grafted cuttings were transferred to the greenhouse for planting under controlled environmental conditions. Table 1 Contents of microbial fertilizers used in the experiment Microbial fertilizer Microbial flora declared by the company on the label Concentration MF-A Total colony count ( Methylobacterium symbioense Sb23) 1.5 x 10 3 cfu/g 2000 ppm MF-B Total colony count ( Glomus intraradices , Glomus mosseae , Glomus aggregatum , Glomus clarum , Glomus monosporus , Glomus deserticola , Glomus brasilianum , Glomus etunicatum , Gigaspora margarita ) 23.5% 2000 ppm MF-C Total number of living organisms ( Glomus fasciculatum ) 60 spores/g 2000 ppm MF-D Total colony count ( Glomus iranicum var. Tenuihypharum ) 5 x 10 6 cfu/ml 2000 ppm MF-E Total colony count ( Pseudomonas fluorescens , Paenibacillus polymyxa , Bacillus megaterium , Pantoea agglomerans ) 1 x 10 7 cfu/ml 2000 ppm MF-F Total colony count ( Bacillus megaterium ) 1 x 10 5 cfu/g 2000 ppm MF-G Total colony count ( Trichoderma harzianum T78) 5 x 10 6 cfu/ml 2000 ppm Microbial fertilizer treatments, as well as positive control, were prepared at a concentration of 2000 ppm. Before planting, the basal 3 cm of each grafted cutting was immersed in the respective solutions for 5 seconds using a rapid dipping technique. Following treatment, all cuttings were planted in polyethylene pots (11 × 11 × 22 cm) filled with a 1:1 mixture of peat and perlite. Throughout the growth period, plants were irrigated once per week with a nutrient solution formulated according to Ollat ( 1998 ) and adjusted to pH 6.5. Each sample received 100 mL of water at 09:30. The greenhouse was maintained under natural photoperiod conditions, and the experiment was conducted in the same greenhouse section in both study years. During the active growth period of 2024 (May–June), the average greenhouse temperature was 25.2 ± 5°C with 50–65% relative humidity. In 2025, average temperature and humidity were 24.8 ± 5°C and 50–60%, respectively. Approximately two months after planting (early July), once sufficient root and shoot development was achieved, measurements were taken for all treatments. Data Collection Each treatment was replicated four times with twelve plants per replicate, resulting in 48 plants per treatment per year. Measurements were performed after the 60-day growth period (1 May–30 June), when grafted vines reached the E-L Stage 15—characterized by approximately eight fully expanded leaves per shoot (Eichhorn and Lorenz 1977 ). Shoot and root lengths were measured in centimeters using a ruler. Root development was evaluated on a 0–4 scale following Damborska ( 1981 ), where 0 indicates the absence of roots and 4 represents well-developed roots completely covering the basal end of the cutting. Plant samples (shoots and cleaned roots) were dried at 60°C for 72 hours, after which dry biomass was recorded (Bourles et al. 2020 ). Fresh and dry weights of shoots and roots were measured with a balance accurate to 0.01 g. Total grafted vine yield (%) was calculated as the proportion of successfully developed, plantable grafted vines relative to the total number of cuttings planted. The resulting plants were classified into first- and second-grade categories. First-grade vines met the standard criteria—including a well-formed graft union, vigorous shoot growth, and a healthy root system—while second-grade vines were viable but did not fully meet these specifications. Statistical Analysis The experiment was arranged in a split-plot design, and analysis of variance (ANOVA) was performed to determine the effects of the studied factors. Differences among means were evaluated using the Least Significant Difference (LSD) test based on Student’s t-test at a significance level of p < 0.05. Furthermore, correlation analysis was performed to identify relationships among the examined parameters. Principal Component Analysis (PCA) was also conducted to exhibit the main components and facilitate the classification of samples. Results The interaction between microbial fertilizer and year was found to be significant for dry root weight, root length, root development, and second-grade seedling yield ( p < 0.05 or p < 0.01). However, this interaction did not significantly affect shoot length, fresh shoot weight, dry shoot weight, fresh root weight, root number, first-grade seedling yield, or total seedling yield ( p > 0.05). The year factor had a highly significant effect on dry root weight (F = 28.5685; p < 0.001) and a significant effect on shoot length (F = 11.6243), fresh root weight (F = 17.4086), and total seedling yield (F = 12.0299) ( p < 0.01). The microbial fertilizer factor showed a strongly significant influence on root number (F = 8.4300), first-grade seedling yield (F = 30.5849), and total seedling yield (F = 33.5485) ( p < 0.001). It also had a significant effect on root number (F = 4.6394) and second-grade seedling yield (F = 4.2075) ( p < 0.01). Detailed statistical results are presented in Table 2 . Table 2 The effects of year and microbial fertilizer, and their interactions on growth and yield parameters of grapevine seedlings Parameters Y (Df) Y (F) Y ( p ) MF (Df) MF (F) MF ( p ) Y × MF (Df) Y × MF (F) Y × MF ( p ) Shoot length 1 11.6243 0.0016* 8 1.1894 0.3325 ns 8 0.5695 0.7956 ns Fresh shoot weight 1 2.3242 0.1361 ns 8 0.5474 0.8128 ns 8 1.2836 0.2826 ns Dry shoot weight 1 3.0025 0.0917 ns 8 0.5851 0.7833 ns 8 0.9898 0.4602 ns Fresh root weight 1 17.4086 0.0002* 8 1.9035 0.0898 ns 8 2.0162 0.0724 ns Dry root weight 1 28.5685 < 0.0001 ** 8 1.7701 0.1158 ns 8 2.4776 0.0298 Root number 1 0.3342 0.5668 ns 8 4.6394 0.0006 * 8 1.2653 0.2918 ns Root length 1 2.1720 0.1492 ns 8 3.6946 0.0031* 8 2.2877 0.0429 Root development 1 2.2069 0.1461 ns 8 1.3966 0.2313 ns 8 3.9138 0.0021* First grade sapling yield 1 3.2768 0.0786 ns 8 30.5849 < 0.0001 ** 8 1.1656 0.3461 ns Second grade sapling yield 1 4.0474 0.0518 ns 8 4.2075 0.0012* 8 3.0087 0.0108 Total sapling yield 1 12.0299 0.0014* 8 33.5485 < 0.0001 ** 8 1.2917 0.2786 ns The analysis was performed using Two-Way ANOVA. Y: Year; MF: Microbial Fertilizer; ns: non-significant; p < 0.05; * p < 0.01; ** p < 0.001 Shoot Length, Fresh Shoot Weight, and Dry Shoot Weight Analysis of shoot length indicated that the year effect was significant (p < 0.01), whereas the effects of microbial fertilizer treatments and the year × treatment interaction were not statistically significant. Fresh and dry shoot weights were not significantly affected by the treatments, year, or their interaction. Shoot length was significantly greater in 2025 (16.33 ± 5.21 cm) than in 2024 (12.11 ± 3.57 cm) (p < 0.01). Although the highest shoot length values were recorded in the MF-A treatment in 2024 (15.33 ± 5.51 cm) and in the MF-E treatment in 2025 (20.67 ± 5.13 cm), these differences were not statistically significant among treatments. Both fresh and dry shoot weights were generally higher in 2025 than in 2024. The highest fresh shoot weight in 2024 was observed in MF-E (6.79 ± 2.23 g), while MF-B produced the highest value in 2025 (8.13 ± 3.33 g). Similarly, the greatest dry shoot weight was recorded in MF-C (3.26 ± 0.97 g) in 2024 and in MF-B (4.13 ± 1.76 g) in 2025 (Table 3 ; Fig. 1 ). Table 3 Effect of microbial fertilizer applications on shoot development Treatments Shoot length (cm) Shoot fresh weight (g) Shoot dry weight (g) 2024 2025 2024 2025 2024 2025 MF-A 15.33 ± 5.51 18.67 ± 7.77 5.96 ± 1.17 7.23 ± 2.39 3.09 ± 0.60 3.63 ± 1.16 MF-B 9.67 ± 1.53 17.33 ± 8.74 4.85 ± 0.93 8.13 ± 3.33 3.21 ± 0.88 4.13 ± 1.76 MF-C 12.67 ± 0.58 16.67 ± 4.51 6.53 ± 1.59 7.37 ± 2.83 3.26 ± 0.97 4.00 ± 1.47 MF-D 13.67 ± 1.53 13.67 ± 5.13 6.32 ± 2.71 5.93 ± 2.90 3.15 ± 1.53 3.06 ± 1.34 MF-E 14.33 ± 4.51 20.67 ± 5.13 6.79 ± 2.23 6.40 ± 0.96 3.25 ± 0.86 3.29 ± 0.46 MF-F 10.67 ± 5.51 18.33 ± 3.51 4.01 ± 1.09 7.83 ± 3.18 2.05 ± 0.64 4.09 ± 1.70 MF-G 11.33 ± 3.79 16.33 ± 5.51 6.17 ± 0.61 6.57 ± 0.42 2.77 ± 0.23 3.67 ± 0.32 Positive control 10.00 ± 2.65 13.67 ± 1.53 5.32 ± 1.12 5.10 ± 0.90 2.95 ± 0.52 2.72 ± 0.39 Negative control 11.33 ± 3.21 11.67 ± 0.58 6.17 ± 0.51 4.67 ± 0.29 3.11 ± 0.12 2.46 ± 0.13 Overall mean 12.11 ± 3.57 b 16.33 ± 5.21 a 5.79 ± 1.51 6.58 ± 2.19 2.98 ± 0.77 3.45 ± 1.12 Values are expressed mean ± SD. Values ​​shown with different letters indicate a significant difference between years according to the LSD test ( p < 0.05). Root Length, Root Number, Root Development, Fresh Root Weight, and Dry Root Weight The microbial fertilizer × year interaction had a statistically significant effect on dry root weight, root length, and root development ( p < 0.05), whereas its effect on fresh root weight and root number was not significant. While the treatments did not significantly influence fresh root weight, the year factor had a significant effect on this parameter ( p < 0.01). The treatments significantly affected root number ( p < 0.01), whereas the year factor did not show a significant influence on root number (Table 4 ). Table 4 Effect of applications on root development Treatments Fresh root weight (g) ** Dry root weight (g) * Root length (cm) * Root development (0–4 scale) ** Root number (piece)** 2024 2025 2024 2025 2024 2025 2024 2025 Combined Mean (2024–2025) MF-A 7.96 ± 3.16 3.73 ± 2.34 3.97 ± 1.03 a 1.66 ± 1.01 d-g 14.0 ± 2.00 b-d 11.3 ± 2.89 c-e 3.7 ± 0.58 ab 3.3 ± 0.58 a-c 15.67 ± 2.88 bc MF-B 3.59 ± 1.80 6.00 ± 0.95 1.36 ± 0.65 e-g 2.64 ± 0.49 a-e 11.3 ± 5.51 c-e 18.7 ± 2.31 ab 3.0 ± 0.58 a-d 4.0 ± 0.00 a 22.83 ± 0.65 a MF-C 6.94 ± 2.43 2.57 ± 1.16 3.59 ± 1.36 ab 1.09 ± 0.51 fg 14.3 ± 5.13 b-d 10.3 ± 2.08 c-e 3.7 ± 0.58 ab 3.0 ± 0.00 a-d 17.67 ± 3.27 b MF-D 7.52 ± 3.19 5.93 ± 0.99 3.70 ± 1.73 ab 2.59 ± 0.43 a-e 15.7 ± 4.04 a-c 21.0 ± 1.00 a 3.0 ± 1.00 a-d 3.0 ± 0,00 a-d 16.33 ± 3.39 bc MF-E 6.86 ± 3.06 2.27 ± 0.64 3.07 ± 1.15 a-d 0.97 ± 0.23 fg 13.7 ± 3.79 b-d 12.0 ± 1.00 c-e 2.7 ± 1.15 b-d 3.3 ± 0.58 a-c 13.33 ± 3.61 bc MF-F 3.87 ± 1.84 1.80 ± 0.79 2.26 ± 1.08 b-f 0.81 ± 0.37 g 12.7 ± 2.08 c-e 13.3 ± 0.58 b-d 3.7 ± 0.58 ab 3.3 ± 0.58 a-c 12.67 ± 2.73 c MF-G 5.59 ± 1.73 3.70 ± 0.52 2.81 ± 0.57 a-d 1.69 ± 0.28 c-g 13.0 ± 5.20 cd 14.3 ± 1.15 b-d 2.3 ± 0.58 c-e 3.3 ± 0.58 a-c 13.33 ± 2.50 bc Positive control 6.02 ± 1.36 4.47 ± 1.25 3.13 ± 0.84 a-c 1.93 ± 0.63 c-g 7.3 ± 1.15 e 13.0 ± 1.00 cd 4.0 ± 0,00 a 1.3 ± 0.58 e 15.67 ± 3.27 bc Negative control 5.33 ± 1.86 4.10 ± 0.95 2.70 ± 1.06 a-e 1.82 ± 0.51 c-g 10.0 ± 5.57 de 9.7 ± 3.21 de 3.3 ± 0.58 a-c 2.0 ± 1.00 de 12.17 ± 3.54 c Overall mean 5.96 ± 2.46 a 3.84 ± 1.74 b 2.95 ± 1.20 1.69 ± 0.77 12.44 ± 4.23 13.74 ± 3.98 3.26 ± 0.90 2.96 ± 0.90 15.52 ± 4.73 Values are expressed mean ± SD. There is a significant difference between the values ​​indicated by different letters. ns: non-significant; * p < 0.05; ** p < 0.01 The overall mean fresh root weight was found to be significantly higher in 2024 (5.96 ± 2.46 g) compared to the overall mean in 2025 (3.84 ± 1.74 g). The highest dry root weight was recorded in the MF-A treatment in 2024 (3.97 ± 1.03 g), while the lowest value was obtained from the MF-F treatment in 2025 (0.81 ± 0.37 g). The highest root length was recorded from the MF-D treatment in 2025 (21.0 ± 1.00 cm), whereas the lowest root length was obtained from the positive control in 2024 (7.3 ± 1.15 cm). In terms of root development score, the positive control in 2024 (4.0) and the MF-B treatment in 2025 (4.0) exhibited the most favorable results, while the positive control in 2025 (1.3) showed the lowest root development level (Fig. 2 ). Regarding the effects of treatments on root number, the highest values were observed under MF-B (22.83 ± 0.65 piece) and MF-C (17.67 ± 3.27 piece), whereas the lowest root numbers were recorded in the MF-F treatment (12.67 ± 2.73 piece) and the negative control (12.17 ± 3.54 piece) (Fig. 3 ). Sapling Yield The interaction between microbial fertilizer and year was found to be statistically significant for second-grade sapling yield ( p < 0.05), whereas its effects on first size sapling yield and total sapling yield were not significant. In contrast, the effects of the treatments on both first size and total sapling yield were highly significant ( p < 0.0001) (Table 5 ; Fig. 4 ). Table 5 Effect of microbial fertilizer applications on first size sapling yield, second size sapling yield and total sapling yield Treatments First grade sapling yield (%) *** Second grade sapling yield (%) * Total sapling yield (%) *** Combined Mean (2024–2025) 2024 2025 Combined Mean (2024–2025) MF-A 46.0 ± 3.18 c 11.7 ± 3.34 d-f 12.3 ± 2.15 c-f 57.9 ± 3.61 d MF-B 57.0 ± 7.05 a 13.3 ± 6.11 c-e 15.0 ± 2.50 c-e 71.2 ± 5.16 ab MF-C 56.3 ± 4.04 a 13.3 ± 3.34 c-e 12.5 ± 2.50 c-f 69.2 ± 4.77 b MF-D 49.0 ± 4.55 bc 10.0 ± 3.33 ef 15.8 ± 1.44 b-d 61.9 ± 3.28 cd MF-E 51.3 ± 1.73 b 11.7 ± 2.89 d-f 7.5 ± 2.50 f 60.8 ± 2.11 cd MF-F 47.0 ± 3.40 bc 13.9 ± 2.55 c-e 21.0 ± 3.61 ab 63.9 ± 5.96 c MF-G 40.7 ± 2.91 d 13.3 ± 3.34 c-e 22.5 ± 4.33 a 58.6 ± 6.51 d Positive control 57.8 ± 6.23 a 17.2 ± 0,96 a-c 15.3 ± 2.52 c-e 74.1 ± 4.67 a Negative control 27.3 ± 4.29 e 15.6 ± 5.09 cd 14.0 ± 1.00 c-e 42.1 ± 2.83 e Values are expressed mean ± SD. There is a significant difference between the values ​​indicated by different letters. ns: non-significant; * p < 0.05; ** p < 0.01; *** p < 0.0001 When the effects of the treatments on first size sapling yield were evaluated, the highest first-grade yields were obtained from the positive control (57.8 ± 6.23%), MF-B (57.0 ± 7.05%), and MF-C (56.3 ± 4.04%) treatments, whereas the lowest first-grade sapling yield was recorded in the negative control (27.3 ± 4.29%). The highest total sapling yield was recorded in the positive control (74.1 ± 4.67%), whereas the lowest total sapling yield was observed in the negative control (42.1 ± 2.83%) (Fig. 4 ). A correlation analysis was performed to evaluate the relationships among the parameters investigated. The strongest positive correlations were detected between fresh root weight and dry root weight (r = 0.96), first-grade sapling yield and second-grade sapling yield (r = 0.96), and fresh shoot weight and dry shoot weight (r = 0.90). The highest negative correlation was found between shoot length and second-grade sapling yield (r = − 0.64) (Fig. 5 ). Among the notable positive correlations, strong relationships were observed between fresh shoot weight and shoot length (r = 0.74), dry shoot weight and root development score (r = 0.72), and dry shoot weight and root number (r = 0.71). Moderate positive correlations were detected between first-grade sapling yield and root number (r = 0.65), fresh shoot weight and root development score (r = 0.63), and total sapling yield and root number (r = 0.62). In contrast, moderate negative correlations were observed between fresh shoot weight and second-grade sapling yield (r = − 0.51), as well as between fresh root weight and second-grade sapling yield (r = − 0.43). To better elucidate the relationships between the treatments and the evaluated parameters, a Principal Component Analysis (PCA) was performed (Fig. 6 ). The first two principal components (Dim1 and Dim2) together accounted for 62.00% of the total variance. Total sapling yield, first-grade sapling yield, dry shoot weight, fresh shoot weight, shoot length, root development score, root number, and root length were positioned on the positive side of the first principal component axis (Dim1), which explained 40.4% of the total variance, whereas second-grade sapling yield was located on the negative side of this axis. Fresh shoot weight, root number, dry shoot weight, first-grade sapling yield, and root development score were found to be closely associated with the MF-B treatment. While total sapling yield was best represented by the MF-C treatment, shoot length was most strongly associated with the MF-A and MF-E treatments. Root length was related to the MF-B and MF-D treatments, whereas second-grade sapling yield was associated with the negative control and MF-G treatments. Fresh root weight and dry root weight were positioned on the positive side of the second principal component axis (Dim2), and both parameters were found to be closely associated with the MF-D treatment (Fig. 6 ). According to the hierarchical clustering heat map, the evaluated parameters were grouped into two main clusters (Fig. 7 ). The first main cluster consisted solely of second-grade sapling yield, while all other parameters were grouped within the second main cluster. This second main cluster was further divided into two sub-clusters. The first sub-cluster included root length, fresh root weight, and dry root weight. The second sub-cluster comprised root number, total sapling yield, first-grade sapling yield, shoot length, root development score, fresh shoot weight, and dry shoot weight. The heat map also classified the treatments into two principal groups. The first main treatment cluster was divided into two sub-clusters, with the first sub-cluster including MF-F, MF-G, and the positive control, and the second sub-cluster consisting of the negative control. The second main treatment cluster was also divided into two sub-clusters, with MF-D forming the first sub-cluster and MF-A, MF-B, MF-C, and MF-E constituting the second sub-cluster. Discussion In the present study, the effects of microbial fertilizer applications on root and shoot development as well as sapling quality parameters were evaluated, and the results clearly demonstrated that microbial formulations exerted particularly pronounced effects on root morphology. The observed increases in root length, dry root weight, and root development score are consistent with previous reports indicating that arbuscular mycorrhizal fungi (AMF) and plant growth-promoting rhizobacteria (PGPR) enhance the root system through mechanisms such as phosphorus mobilization in the rhizosphere, stimulation of lateral root formation, and the production of hormone-like compounds (Azcón-Aguilar and Barea 1997 ; Ortiz-Castro et al. 2009 ; Cely et al. 2016 ). In particular, the fact that the MF-B, MF-C, and MF-D treatments exhibited performances comparable to the positive control strongly supports the notion that microbial inoculants play a powerful physiological and morphological stimulatory role in root tissues. The statistically significant “year × treatment” interaction observed in this study further indicates that microbial efficacy is highly sensitive to environmental conditions. Previous studies have shown that soil moisture, temperature, pH, and nutrient availability are decisive factors regulating AMF activity and root colonization. It has been widely reported that the colonization rate and functional efficiency of AMF are strongly influenced by interannual environmental variability (Ortas 2010 ; Powell and Rillig 2018 ; Wahab et al. 2023 ; Chen et al. 2023 ). Conversely, some studies have suggested that AMF can provide relatively stable growth promotion regardless of year or environmental fluctuations (Bhardwaj et al. 2024 ). Therefore, the year-dependent differences observed in root weight and root development level in the present study confirm the context-dependent and variable performance of microbial fertilizers. The limited and mostly non-significant effects of microbial treatments on shoot length and shoot biomass parameters are also in agreement with the existing literature. Numerous studies have reported that AMF influence aerial plant parts with lower sensitivity compared to the root zone (Geisen et al. 2022 ). However, in contrast, several investigations have shown that mycorrhizal applications can significantly enhance shoot growth under certain conditions (Wu et al. 2022 ; Chandrasekaran 2022 ; Xu et al. 2024 ). These discrepancies suggest that microbial effectiveness is strongly dependent on factors such as plant species, inoculum density, root infection rate, and growing conditions. In this study, the significant effects of the “microbial fertilizer × year” interaction on dry root weight, root length, and root development score ( p < 0.05) indicate that the response of grapevine roots to microbial applications is strongly modulated by environmental conditions and interannual variability. This finding is consistent with previous studies reporting that variations in soil moisture, temperature, and microbial activity across years can substantially influence root–microorganism interactions (Torres et al. 2021 ; Cardinale et al. 2022 ). In particular, the higher fresh root weight observed in 2024 (5.96 ± 2.46 g) supports the findings of Lailheugue et al. ( 2024 ), who reported that environmental conditions play a decisive role in determining AMF-mediated root metabolism and colonization success. The highest dry root weight (3.97 ± 1.03 g) was obtained from the MF-A treatment containing the Methylobacterium symbioticum SB23 strain in 2024. This result agrees with previous findings demonstrating that M. symbioticum produces indole-3-acetic acid (IAA), which stimulates lateral root formation and enhances root–soil contact (Patten and Glick 2002 ), thereby positively affecting root biomass (Bolla et al. 2025 ). In contrast, the low dry root weight observed under the MF-F treatment in 2025 suggests that the effects of microbial inoculants on root development may vary depending on plant genotype, inoculum characteristics, and seasonal conditions. Moreover, the highest root length recorded in 2025 under the MF-D treatment (21.0 ± 1.00 cm) may be attributed to differential microbial responsiveness of the rootstock used (Sportes et al. 2023 ). With respect to root development score, the fact that both the positive control in 2024 and the MF-B treatment in 2025 achieved the highest score (4.0) indicates that microbial fertilizers can, under certain conditions, improve root morphology to a level comparable with that of natural microbial communities. However, the marked decline observed in the positive control in 2025 (1.3) highlights the pronounced role of the year factor in determining root development performance and is consistent with the literature emphasizing the significance of year-based variability in viticulture (Romero et al. 2025 ). In this study, root number was significantly affected by the treatments ( p < 0.01), whereas the effect of the year factor was not significant, indicating that microbial fertilizer applications can directly stimulate root branching independently of interannual variation. The highest root numbers obtained under the MF-B (22.83 ± 0.65) and MF-C (17.67 ± 3.27) treatments are in accordance with previous findings demonstrating that AMF and PGPR consortia enhance root branching (Trouvelot et al. 2015 ; Torres et al. 2021 ). Conversely, the low root number observed in the MF-F treatment suggests that not all microbial inoculant variants exert positive effects on root architecture and that root–microorganism compatibility is a critical determinant of success. Overall, these findings demonstrate that microbial fertilizer applications have a strong potential to enhance root architecture components such as root biomass, root length, root development score, and root number in Narince grapevine saplings; however, the magnitude of these effects varies depending on year conditions and application type. The literature reports that the sensitivity of grapevine root development to microbial inoculation is strongly influenced by environmental factors and the composition of the applied microbial populations (Goddard et al. 2021 ; Lailheugue et al. 2024 ; Moukarzel et al. 2024 ), and the results of the present study are in close agreement with these observations. In general, microbial fertilizer applications exerted positive effects on root architecture and sapling quality class in grapevine nursery production, whereas the responses of shoot-related parameters remained limited and the overall effectiveness of the treatments varied between years. These results suggest that microbial fertilizers may be considered as an alternative or complementary biological input to chemical fertilizers under nursery conditions; however, achieving optimal efficacy requires careful selection of environmental conditions, microbial diversity, and application strategies. Conclusion This study comprehensively evaluated the effects of different microbial fertilizer formulations on container-grown grapevine sapling production in the Narince/110 R combination over a two-year experimental period. The findings revealed that microbial fertilizers exerted pronounced effects particularly on root morphology, sapling quality, and total sapling yield. The significant “microbial fertilizer × year” interaction for root length, dry root weight, and root development score demonstrated that the performance of these applications is highly sensitive to environmental conditions. In contrast, the effects of the treatments on shoot length, fresh shoot weight, and dry shoot weight were generally limited. The MF-B, MF-C, and MF-D treatments emerged as the most effective in terms of root system performance and sapling quality; notably, the MF-B treatment exhibited superior performance with respect to root number, root development score, and first-grade sapling yield. Although the positive control (IBA) produced the highest total sapling yield in both years, the fact that several microbial fertilizer treatments (MF-B and MF-C) achieved results comparable to this application indicate that biological inputs may serve as an alternative or complementary option to chemical hormone-based treatments in nursery production. Correlation and PCA analyses confirmed that sapling yield is largely associated with root biomass and root functionality and clearly demonstrated that the primary mode of action of microbial fertilizers is mediated through the root system. This finding supports the potential of microbial fertilizers to improve sapling quality by enhancing root efficiency, particularly under limited volume growing media conditions. Overall, this study demonstrates that microbial fertilizers represent a viable biological input for container-grown grapevine sapling production and can make substantial contributions to improving sapling yield and root system development. However, the variation in application performance between years highlights the need for further large-scale studies under different ecological regions, rootstock–cultivar combinations and growing conditions. Such studies are essential for the standardization and widespread adoption of microbial fertilizers as a biological production input in viticulture. Tablo 6 Effects of Microbial Fertilizer Treatments and Control Groups Treatment Component Mode of Action Consistence with the Findings of the Present Study MF-A Methylobacterium symbioense Endophytic nitrogen fixation and phytohormone production Enhanced shoot growth and increased fresh root weight MF-B Glomus spp . + Humic acid, vitamins, and amino acids Root colonization, enhanced nutrient uptake, and hormone-like effects Highest root number and sapling yield, with improved shoot growth MF-C Glomus fasciculatum Phosphorus uptake and root support Balanced shoot and root development with good sapling yield MF-D Glomus iranicum Root elongation and enhanced stress tolerance Longest root length, high root biomass, and good sapling yield MF-E PGPR consortium ( Pseudomonas , Bacillus , Paenibacillus ) Phytohormone production and phosphate solubilization High shoot development with low dry root weight MF-F Bacillus megaterium Phosphate solubilization and root stimulation High proportion of second size saplings, with limited shoot and root development MF-G Trichoderma harzianum Biocontrol and indirect growth promotion Moderate root development with a high proportion of second size saplings Positive control Synthetic auxin (2000 ppm) Rapid root stimulation High root development score, but lower biomass and sapling yield compared to microbial fertilizers Negative control No application - Low values across all parameters Declarations Acknowledgements The authors would like to thank Tokat Gaziosmanpaşa University, Faculty of Agriculture. Funding The research work was self-funded. Authors and Affiliations Department of Horticulture, Faculty of Agricultural, University of Tokat Gaziosmanpaşa University, Turkey Adem Yağcı Tokat Food Control Laboratory Directorate, Tokat, Turkey İsa Hazar Department of Horticulture, Faculty of Agriculture, Tokat Gaziosmanpaşa University, Turkey Rüstem Cangi Department of Horticulture, Faculty of Agriculture, Kocaeli University, Turkey Selda Daler Authors’ Contributions All authors contributed to the study conception and design. Material preparation, experimental conduction, data collection by Adem Yağcı, Rüstem Cangi, Selda Daler, İsa Hazar. The manuscript was written by İsa Hazar and all authors approved the final manuscript. Corresponding author Correspondence to İsa Hazar Ethics approval and consent to participate Not applicable. This study did not involve human participants, human data, or animals. Consent for publication Not applicable. Conflicts of Interest İ. Hazar, A. Yağcı, S. 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Supplementary Files GA.png Graphic Abstract MicrobialFertilizerSupplementarydata.docx Cite Share Download PDF Status: Posted Version 1 posted 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. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-8475362","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":569726891,"identity":"ea082516-f81d-446a-b054-f89881ee058a","order_by":0,"name":"İsa 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1","display":"","copyAsset":false,"role":"figure","size":53747,"visible":true,"origin":"","legend":"\u003cp\u003eImpact of each treatment on the shoot length, fresh shoot weight, and dry shoot weight of Narince grapevine saplings\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-8475362/v1/afb804268a1ed73512b2acc3.png"},{"id":99793564,"identity":"aa69b4f5-1ccb-483d-8f49-b8ae1eb7d93c","added_by":"auto","created_at":"2026-01-08 13:31:52","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":69221,"visible":true,"origin":"","legend":"\u003cp\u003eEffects of microbial fertilizer applications on root length, root development level, fresh root weight and dry root weight in grapevine saplings\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-8475362/v1/025f9cca28d132563b7080ad.png"},{"id":99793505,"identity":"4489f44c-efd7-4800-a7ec-19884bd6580e","added_by":"auto","created_at":"2026-01-08 13:31:42","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":30203,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of microbial fertilizer applications on root number in grapevine seedlings\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-8475362/v1/055ef940ccdb9c0a21e77016.png"},{"id":99618498,"identity":"1d8cdedf-60b5-4d43-a314-65f42fcfe2de","added_by":"auto","created_at":"2026-01-06 13:53:04","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":36319,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of microbial fertilizer applications on first size sapling yield and total sapling yield\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-8475362/v1/80e7777c25350c72b5076988.png"},{"id":99618508,"identity":"42520371-3a24-497e-b7a7-a7a27c1f28b3","added_by":"auto","created_at":"2026-01-06 13:53:05","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":92894,"visible":true,"origin":"","legend":"\u003cp\u003ePearson correlation analysis showing the relationships between shoot and root development and yield parameters of Narince grapevine saplings. SL: Shoot length; FSW: Fresh shoot weight; DSW: Dry shoot weight; RN: Root number; RL: Root length; RD: Root development; FRW: Fresh root weight; DRW: Dry root weight; FSSY: First grade sapling yield; SSSY: Second grade sapling yield; TSY: Total sapling yield.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-8475362/v1/81653058b79646501cc78ca2.png"},{"id":99793879,"identity":"348654bb-c14c-4436-afae-b492e6f49bce","added_by":"auto","created_at":"2026-01-08 13:33:25","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":39588,"visible":true,"origin":"","legend":"\u003cp\u003ePrincipal Component Analysis (PCA) visualizing the relationships between shoot and root development and yield parameters of Narince grapevine saplings. SL: Shoot length; FSW: Fresh shoot weight; DSW: Dry shoot weight; RN: Root number; RL: Root length; RD: Root development; FRW: Fresh root weight; DRW: Dry root weight; FSSY: First grade sapling yield; SSSY: Second grade sapling yield; TSY: Total sapling yield. PC: Positive control; NC: Negative control.\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-8475362/v1/f92d3caaea359a1787b38737.png"},{"id":99793872,"identity":"4a7585d1-3045-4f5c-b6cd-3572fa7c4eb7","added_by":"auto","created_at":"2026-01-08 13:33:24","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":43201,"visible":true,"origin":"","legend":"\u003cp\u003eHierarchical clustering heat map represents the effects of applications on shoot and root development and yield of Narince grapevine saplings. SL: Shoot length; FSW: Fresh shoot weight; DSW: Dry shoot weight; RN: Root number; RL: Root length; RD: Root development; FRW: Fresh root weight; DRW: Dry root weight; FSSY: First grade sapling yield; SSSY: Second grade sapling yield; TSY: Total sapling yield; PC: Positive control; NC: Negative control.\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-8475362/v1/2f8efe65e1909791decca7c3.png"},{"id":102910231,"identity":"26788d42-d9e8-4143-af58-8a4122a66d41","added_by":"auto","created_at":"2026-02-18 09:57:51","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1648192,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8475362/v1/f891fae4-409d-4e43-8da7-a1766325cf3d.pdf"},{"id":99793046,"identity":"9fe6d293-32aa-40a2-8999-0db397844be2","added_by":"auto","created_at":"2026-01-08 13:30:55","extension":"png","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":371726,"visible":true,"origin":"","legend":"\u003cp\u003eGraphic Abstract\u003c/p\u003e","description":"","filename":"GA.png","url":"https://assets-eu.researchsquare.com/files/rs-8475362/v1/a572408fa527d90afa0345ef.png"},{"id":99794746,"identity":"f515e48b-18f8-4ab3-9bb9-88cf971a1d67","added_by":"auto","created_at":"2026-01-08 13:36:10","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":565789,"visible":true,"origin":"","legend":"","description":"","filename":"MicrobialFertilizerSupplementarydata.docx","url":"https://assets-eu.researchsquare.com/files/rs-8475362/v1/d384351acfec24d6c1bc1ff5.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Use of Microbial Fertilizers in Grapevine Seedling Production","fulltext":[{"header":"Introduction","content":"\u003cp\u003eGrapevine (\u003cem\u003eVitis vinifera\u003c/em\u003e L.) is one of the most economically and culturally important crops in T\u0026uuml;rkiye, where nursery production constitutes the structural foundation of vineyard establishment (Korkutal et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). Nevertheless, grapevine nursery production continues to face major challenges, including limited availability of certified, disease-free and true-to-type planting material, which directly affects production efficiency and cost (Bahar et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Durmaz et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Uysal and Ateş \u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). In addition, structural limitations within the Turkish nursery sector, such as production capacity and marketing conditions, further constrain producer competitiveness (Cangi et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Consequently, the demand for innovative and sustainable biological inputs has increased.\u003c/p\u003e \u003cp\u003eAmong the main factors influencing seedling yield and quality, fertilization practices play a central role. Environmental concerns associated with conventional fertilization and its long-term negative impacts on soil health have accelerated the adoption of eco-friendly alternatives. In this context, microbial fertilizers and plant biostimulants have attracted considerable attention due to their potential to enhance plant growth and productivity. Biostimulants are generally classified as microbial and non-microbial, with microbial biostimulants primarily consisting of arbuscular mycorrhizal fungi (AMF), plant growth\u0026ndash;promoting rhizobacteria (PGPR) and \u003cem\u003eTrichoderma\u003c/em\u003e spp. (Van Loon \u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Rouphael and Colla \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Tsvetkov et al. \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003cp\u003ePlant growth is largely governed by root\u0026ndash;soil interactions, in which roots and microorganisms engage in both nutrient competition and mutualistic symbiosis (Belal et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Martin and Van der Heijden \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Most terrestrial plant species form associations with mycorrhizal fungi that enhance nutrient and water uptake in exchange for plant-derived carbon (Sandal Erzurumlu and Kara 2014; Agnolucci et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Moukarzel et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eMicrobial biostimulants have been extensively investigated in grapevine propagation. Mycorrhizal preparations have been evaluated in grapevine cuttings, rootstocks and grafted seedlings (Gendiah 1991; Kara and \u0026Ouml;zdemir \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Kavak \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Kara et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Eftekhari et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Eroğlu and \u0026Ccedil;elik \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2013\u003c/span\u003elı\u0026ccedil; and Cangi 2019), as well as under vineyard conditions, with variable effects on growth and nutrient acquisition (Camprub\u0026iacute; et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). Beneficial microorganisms commonly used in biofertilizers include \u003cem\u003eBacillus\u003c/em\u003e, \u003cem\u003eAzotobacter\u003c/em\u003e, \u003cem\u003eTrichoderma\u003c/em\u003e, \u003cem\u003eRhizobium\u003c/em\u003e, \u003cem\u003eAzospirillum\u003c/em\u003e and \u003cem\u003eSaccharomyces\u003c/em\u003e spp., among which \u003cem\u003eTrichoderma\u003c/em\u003e spp. are particularly well studied due to their combined roles in biocontrol and plant growth promotion (Calvo et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Di Marco and Osti \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Cangi et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Numerous studies have demonstrated that \u003cem\u003eTrichoderma\u003c/em\u003e spp. stimulate plant growth, improve root and shoot development, enhance yield, increase tolerance to abiotic stresses, promote nutrient use efficiency and enhance photosynthetic activity (Yedidia et al. \u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; Harman \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Woo et al. \u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Di Marco and Osti \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Tsvetkov et al. \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Mahmood \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Korkutal et al. \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eSimilarly, PGPR applications on grapevine rootstocks have been shown to improve vegetative growth, mineral nutrition and chlorophyll content while reducing chemical fertilizer requirements (Sabir et al. \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2012\u003c/span\u003e), thereby supporting sustainable production systems (Rouphael and Colla \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Plant growth\u0026ndash;promoting microorganisms enhance plant performance through phytohormone production, improved mineral acquisition, biological nitrogen fixation and phosphorus solubilization, although their effectiveness depends strongly on microbial composition, application rate, plant genotype and environmental conditions (Lucy et al. \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Etesami and Maheshwari \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Lopes et al. \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Etesami and Glick \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Khan et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Nagrale et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eTherefore, systematic evaluation of microbial fertilizer formulations in economically important crops such as grapevine, particularly at the nursery stage, remains essential. The objective of this study was to determine the effects of different microbial fertilizers on shoot and root development and overall seedling quality in container-grown grafted grapevine production. We hypothesized that the application of different microbial fertilizer formulations would significantly enhance root system development and sapling quality of grafted grapevine (\u003cem\u003eVitis vinifera\u003c/em\u003e L. cv. Narince/110 R) compared with control groups (positive and negative controls), and that the magnitude of these effects would vary among microbial consortia and across years due to environmental modulation of root\u0026ndash;microbe interactions.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eExperimental Site and Plant Material\u003c/h2\u003e \u003cp\u003eThe experiment was conducted over the 2024 and 2025 propagation seasons in the grapevine seedling facilities and greenhouse units of the Agricultural Practice and Research Center at Tokat Gaziosmanpaşa University (TOGU), T\u0026uuml;rkiye. The plant material consisted of the wine grape cultivar Narince grafted onto the 110 R rootstock (V. \u003cem\u003eberlandieri\u003c/em\u003e Ress\u0026eacute;guier No.2 \u0026times; \u003cem\u003eV. rupestris\u003c/em\u003e Martin 110 Richter), a genotype widely recognized for its tendency to root poorly (Howell \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e1987\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eRootstock cuttings were supplied by the Manisa Viticulture Research Institute, whereas scion cuttings were collected from the TOGU research vineyard. Rootstock cuttings were prepared from the well-matured mid-sections of one-year-old dormant shoots in accordance with the approach described by Roux (1988). Following TS-4027 criteria, 110 R cuttings were selected at 30\u0026ndash;40 cm in length and 8\u0026ndash;12 mm in diameter. The \u003cem\u003eNarince\u003c/em\u003e scion shoots were roughly 100 cm long and averaged 15 buds. All cuttings were immersed in a fungicide solution (1 kg Switch 62.5 WG per m\u0026sup3; water), sealed in polyethylene bags, and maintained at +\u0026thinsp;4\u0026deg;C with 80\u0026ndash;95% relative humidity for approximately 60 days until grafting (Ağaoğlu and \u0026Ccedil;elik \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e1978\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eGrafting Procedures and Callusing Conditions\u003c/h3\u003e\n\u003cp\u003eFive days before grafting, cuttings were removed from cold storage and allowed to acclimate gradually to room temperature. To compensate for moisture depletion during storage, rootstock and scion cuttings were soaked in fungicide-amended water for 24 and 48 hours, respectively (Sucu et al. \u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Thermotherapy was then applied to reduce potential pathogen load: scions were treated for 15 minutes at 50\u0026deg;C, and rootstocks for 30 minutes (Sağlam \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eGrafting was performed using a pedal-operated Ω-type grafting machine. Immediately afterward, the upper 5\u0026ndash;7 cm of each grafted cutting, including the graft union, was coated with paraffin at 74\u0026ndash;76\u0026deg;C for 1\u0026ndash;2 seconds. This step helped prevent dehydration, reduced contamination risk, and supported the initial callus formation process (Richards \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e1976\u003c/span\u003e). To avoid any heat-related bud injury, each cutting was briefly cooled in water for 1\u0026ndash;2 seconds (K\u0026uuml;\u0026ccedil;\u0026uuml;kyumuk 2009). Fungicide-moistened pine sawdust (30 g Switch 62.5 WG in 15 L water) was used as the bedding medium during callusing.\u003c/p\u003e \u003cp\u003eGrafted cuttings were placed in plastic trays layered with sawdust and transferred to a callusing room for a 21-day period. During this time, temperature and humidity gradually decreased:\u003c/p\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003e27\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C and 80\u0026ndash;85% RH for the first 3 days,\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003e25\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C and 80\u0026ndash;82% RH for the following 15 days,\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003e23\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C and 75\u0026ndash;80% RH for the final 3 days (Aslan et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2015\u003c/span\u003e).\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003cp\u003eFollowing callusing, the cuttings were slowly acclimatized for about one week. Sawdust was removed using pressurized air, and a second paraffin dip (80\u0026ndash;82\u0026deg;C) was applied to maintain callus moisture. Prior to planting, the basal 10 cm of each cutting was soaked in water for 3 days.\u003c/p\u003e\n\u003ch3\u003eMicrobial Fertilizer Treatments and Planting\u003c/h3\u003e\n\u003cp\u003eThe experimental design included seven microbial fertilizer treatments along with a positive control (Indole-3-butyric acid) and an untreated negative control. The microbial compositions of the fertilizers used in the study are presented in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. After hydration and preparation steps were completed, the grafted cuttings were transferred to the greenhouse for planting under controlled environmental conditions.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eContents of microbial fertilizers used in the experiment\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMicrobial fertilizer\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMicrobial flora declared by the company on the label\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eConcentration\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTotal colony count (\u003cem\u003eMethylobacterium symbioense\u003c/em\u003e Sb23) 1.5 x 10\u003csup\u003e3\u003c/sup\u003e cfu/g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2000 ppm\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-B\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTotal colony count (\u003cem\u003eGlomus intraradices\u003c/em\u003e, \u003cem\u003eGlomus mosseae\u003c/em\u003e, \u003cem\u003eGlomus aggregatum\u003c/em\u003e, \u003cem\u003eGlomus clarum\u003c/em\u003e, \u003cem\u003eGlomus monosporus\u003c/em\u003e, \u003cem\u003eGlomus deserticola\u003c/em\u003e, \u003cem\u003eGlomus brasilianum\u003c/em\u003e, \u003cem\u003eGlomus etunicatum\u003c/em\u003e, \u003cem\u003eGigaspora margarita\u003c/em\u003e) 23.5%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2000 ppm\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTotal number of living organisms (\u003cem\u003eGlomus fasciculatum\u003c/em\u003e) 60 spores/g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2000 ppm\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-D\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTotal colony count (\u003cem\u003eGlomus iranicum\u003c/em\u003e var. \u003cem\u003eTenuihypharum\u003c/em\u003e) 5 x 10\u003csup\u003e6\u003c/sup\u003e cfu/ml\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2000 ppm\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-E\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTotal colony count (\u003cem\u003ePseudomonas fluorescens\u003c/em\u003e, \u003cem\u003ePaenibacillus polymyxa\u003c/em\u003e, \u003cem\u003eBacillus megaterium\u003c/em\u003e, \u003cem\u003ePantoea agglomerans\u003c/em\u003e) 1 x 10\u003csup\u003e7\u003c/sup\u003e cfu/ml\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2000 ppm\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-F\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTotal colony count (\u003cem\u003eBacillus megaterium\u003c/em\u003e) 1 x 10\u003csup\u003e5\u003c/sup\u003e cfu/g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2000 ppm\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-G\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTotal colony count (\u003cem\u003eTrichoderma harzianum\u003c/em\u003e T78) 5 x 10\u003csup\u003e6\u003c/sup\u003e cfu/ml\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2000 ppm\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eMicrobial fertilizer treatments, as well as positive control, were prepared at a concentration of 2000 ppm. Before planting, the basal 3 cm of each grafted cutting was immersed in the respective solutions for 5 seconds using a rapid dipping technique. Following treatment, all cuttings were planted in polyethylene pots (11 \u0026times; 11 \u0026times; 22 cm) filled with a 1:1 mixture of peat and perlite. Throughout the growth period, plants were irrigated once per week with a nutrient solution formulated according to Ollat (\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e1998\u003c/span\u003e) and adjusted to pH 6.5. Each sample received 100 mL of water at 09:30. The greenhouse was maintained under natural photoperiod conditions, and the experiment was conducted in the same greenhouse section in both study years. During the active growth period of 2024 (May\u0026ndash;June), the average greenhouse temperature was 25.2\u0026thinsp;\u0026plusmn;\u0026thinsp;5\u0026deg;C with 50\u0026ndash;65% relative humidity. In 2025, average temperature and humidity were 24.8\u0026thinsp;\u0026plusmn;\u0026thinsp;5\u0026deg;C and 50\u0026ndash;60%, respectively. Approximately two months after planting (early July), once sufficient root and shoot development was achieved, measurements were taken for all treatments.\u003c/p\u003e\n\u003ch3\u003eData Collection\u003c/h3\u003e\n\u003cp\u003eEach treatment was replicated four times with twelve plants per replicate, resulting in 48 plants per treatment per year. Measurements were performed after the 60-day growth period (1 May\u0026ndash;30 June), when grafted vines reached the E-L Stage 15\u0026mdash;characterized by approximately eight fully expanded leaves per shoot (Eichhorn and Lorenz \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e1977\u003c/span\u003e). Shoot and root lengths were measured in centimeters using a ruler. Root development was evaluated on a 0\u0026ndash;4 scale following Damborska (\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e1981\u003c/span\u003e), where 0 indicates the absence of roots and 4 represents well-developed roots completely covering the basal end of the cutting. Plant samples (shoots and cleaned roots) were dried at 60\u0026deg;C for 72 hours, after which dry biomass was recorded (Bourles et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Fresh and dry weights of shoots and roots were measured with a balance accurate to 0.01 g. Total grafted vine yield (%) was calculated as the proportion of successfully developed, plantable grafted vines relative to the total number of cuttings planted. The resulting plants were classified into first- and second-grade categories. First-grade vines met the standard criteria\u0026mdash;including a well-formed graft union, vigorous shoot growth, and a healthy root system\u0026mdash;while second-grade vines were viable but did not fully meet these specifications.\u003c/p\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eStatistical Analysis\u003c/h2\u003e \u003cp\u003eThe experiment was arranged in a split-plot design, and analysis of variance (ANOVA) was performed to determine the effects of the studied factors. Differences among means were evaluated using the Least Significant Difference (LSD) test based on Student\u0026rsquo;s t-test at a significance level of \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05. Furthermore, correlation analysis was performed to identify relationships among the examined parameters. Principal Component Analysis (PCA) was also conducted to exhibit the main components and facilitate the classification of samples.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003eThe interaction between microbial fertilizer and year was found to be significant for dry root weight, root length, root development, and second-grade seedling yield (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05 or \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.01). However, this interaction did not significantly affect shoot length, fresh shoot weight, dry shoot weight, fresh root weight, root number, first-grade seedling yield, or total seedling yield (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05). The year factor had a highly significant effect on dry root weight (F\u0026thinsp;=\u0026thinsp;28.5685; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001) and a significant effect on shoot length (F\u0026thinsp;=\u0026thinsp;11.6243), fresh root weight (F\u0026thinsp;=\u0026thinsp;17.4086), and total seedling yield (F\u0026thinsp;=\u0026thinsp;12.0299) (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.01). The microbial fertilizer factor showed a strongly significant influence on root number (F\u0026thinsp;=\u0026thinsp;8.4300), first-grade seedling yield (F\u0026thinsp;=\u0026thinsp;30.5849), and total seedling yield (F\u0026thinsp;=\u0026thinsp;33.5485) (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001). It also had a significant effect on root number (F\u0026thinsp;=\u0026thinsp;4.6394) and second-grade seedling yield (F\u0026thinsp;=\u0026thinsp;4.2075) (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.01). Detailed statistical results are presented in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe effects of year and microbial fertilizer, and their interactions on growth and yield parameters of grapevine seedlings\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"10\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eParameters\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eY\u003c/p\u003e \u003cp\u003e(Df)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eY\u003c/p\u003e \u003cp\u003e(F)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eY\u003c/p\u003e \u003cp\u003e(\u003cem\u003ep\u003c/em\u003e)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMF\u003c/p\u003e \u003cp\u003e(Df)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMF\u003c/p\u003e \u003cp\u003e(F)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMF\u003c/p\u003e \u003cp\u003e(\u003cem\u003ep\u003c/em\u003e)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eY \u0026times; MF\u003c/p\u003e \u003cp\u003e(Df)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eY \u0026times; MF\u003c/p\u003e \u003cp\u003e(F)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003eY \u0026times; MF\u003c/p\u003e \u003cp\u003e(\u003cem\u003ep\u003c/em\u003e)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eShoot length\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e11.6243\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.0016*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.1894\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.3325 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e0.5695\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0.7956 ns\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFresh shoot weight\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.3242\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.1361 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.5474\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.8128 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e1.2836\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0.2826 ns\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDry shoot weight\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3.0025\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.0917 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.5851\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.7833 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e0.9898\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0.4602 ns\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFresh root weight\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e17.4086\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.0002*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.9035\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.0898 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e2.0162\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0.0724 ns\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDry root weight\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e28.5685\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.0001 **\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.7701\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.1158 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e2.4776\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0.0298\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRoot number\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.3342\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.5668 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e4.6394\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.0006 *\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e1.2653\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0.2918 ns\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRoot length\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.1720\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.1492 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e3.6946\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.0031*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e2.2877\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0.0429\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRoot development\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.2069\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.1461 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.3966\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.2313 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e3.9138\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0.0021*\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFirst grade sapling yield\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3.2768\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.0786 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e30.5849\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.0001 **\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e1.1656\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0.3461 ns\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSecond grade sapling yield\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e4.0474\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.0518 ns\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e4.2075\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.0012*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e3.0087\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0.0108\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTotal sapling yield\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e12.0299\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.0014*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e33.5485\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.0001 **\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e1.2917\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0.2786 ns\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"10\"\u003eThe analysis was performed using Two-Way ANOVA. Y: Year; MF: Microbial Fertilizer; ns: non-significant; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05; *\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.01; **\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e\n\u003ch3\u003eShoot Length, Fresh Shoot Weight, and Dry Shoot Weight\u003c/h3\u003e\n\u003cp\u003eAnalysis of shoot length indicated that the year effect was significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.01), whereas the effects of microbial fertilizer treatments and the year \u0026times; treatment interaction were not statistically significant. Fresh and dry shoot weights were not significantly affected by the treatments, year, or their interaction.\u003c/p\u003e \u003cp\u003eShoot length was significantly greater in 2025 (16.33\u0026thinsp;\u0026plusmn;\u0026thinsp;5.21 cm) than in 2024 (12.11\u0026thinsp;\u0026plusmn;\u0026thinsp;3.57 cm) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.01). Although the highest shoot length values were recorded in the MF-A treatment in 2024 (15.33\u0026thinsp;\u0026plusmn;\u0026thinsp;5.51 cm) and in the MF-E treatment in 2025 (20.67\u0026thinsp;\u0026plusmn;\u0026thinsp;5.13 cm), these differences were not statistically significant among treatments.\u003c/p\u003e \u003cp\u003eBoth fresh and dry shoot weights were generally higher in 2025 than in 2024. The highest fresh shoot weight in 2024 was observed in MF-E (6.79\u0026thinsp;\u0026plusmn;\u0026thinsp;2.23 g), while MF-B produced the highest value in 2025 (8.13\u0026thinsp;\u0026plusmn;\u0026thinsp;3.33 g). Similarly, the greatest dry shoot weight was recorded in MF-C (3.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.97 g) in 2024 and in MF-B (4.13\u0026thinsp;\u0026plusmn;\u0026thinsp;1.76 g) in 2025 (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e; Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eEffect of microbial fertilizer applications on shoot development\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatments\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eShoot length (cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003eShoot fresh weight (g)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003eShoot dry weight (g)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2024\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2025\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2024\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2025\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2024\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2025\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e15.33\u0026thinsp;\u0026plusmn;\u0026thinsp;5.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e18.67\u0026thinsp;\u0026plusmn;\u0026thinsp;7.77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.96\u0026thinsp;\u0026plusmn;\u0026thinsp;1.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e7.23\u0026thinsp;\u0026plusmn;\u0026thinsp;2.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3.63\u0026thinsp;\u0026plusmn;\u0026thinsp;1.16\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-B\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e9.67\u0026thinsp;\u0026plusmn;\u0026thinsp;1.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e17.33\u0026thinsp;\u0026plusmn;\u0026thinsp;8.74\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.85\u0026thinsp;\u0026plusmn;\u0026thinsp;0.93\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e8.13\u0026thinsp;\u0026plusmn;\u0026thinsp;3.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e4.13\u0026thinsp;\u0026plusmn;\u0026thinsp;1.76\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e12.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e16.67\u0026thinsp;\u0026plusmn;\u0026thinsp;4.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6.53\u0026thinsp;\u0026plusmn;\u0026thinsp;1.59\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e7.37\u0026thinsp;\u0026plusmn;\u0026thinsp;2.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e4.00\u0026thinsp;\u0026plusmn;\u0026thinsp;1.47\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-D\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e13.67\u0026thinsp;\u0026plusmn;\u0026thinsp;1.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e13.67\u0026thinsp;\u0026plusmn;\u0026thinsp;5.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6.32\u0026thinsp;\u0026plusmn;\u0026thinsp;2.71\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5.93\u0026thinsp;\u0026plusmn;\u0026thinsp;2.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.15\u0026thinsp;\u0026plusmn;\u0026thinsp;1.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3.06\u0026thinsp;\u0026plusmn;\u0026thinsp;1.34\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-E\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14.33\u0026thinsp;\u0026plusmn;\u0026thinsp;4.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e20.67\u0026thinsp;\u0026plusmn;\u0026thinsp;5.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6.79\u0026thinsp;\u0026plusmn;\u0026thinsp;2.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.46\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-F\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10.67\u0026thinsp;\u0026plusmn;\u0026thinsp;5.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e18.33\u0026thinsp;\u0026plusmn;\u0026thinsp;3.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.01\u0026thinsp;\u0026plusmn;\u0026thinsp;1.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e7.83\u0026thinsp;\u0026plusmn;\u0026thinsp;3.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.64\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e4.09\u0026thinsp;\u0026plusmn;\u0026thinsp;1.70\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-G\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e11.33\u0026thinsp;\u0026plusmn;\u0026thinsp;3.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e16.33\u0026thinsp;\u0026plusmn;\u0026thinsp;5.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6.57\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.77\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePositive control\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10.00\u0026thinsp;\u0026plusmn;\u0026thinsp;2.65\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e13.67\u0026thinsp;\u0026plusmn;\u0026thinsp;1.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.32\u0026thinsp;\u0026plusmn;\u0026thinsp;1.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.95\u0026thinsp;\u0026plusmn;\u0026thinsp;0.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.39\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNegative control\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e11.33\u0026thinsp;\u0026plusmn;\u0026thinsp;3.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e11.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOverall mean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e12.11\u0026thinsp;\u0026plusmn;\u0026thinsp;3.57 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e16.33\u0026thinsp;\u0026plusmn;\u0026thinsp;5.21 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.79\u0026thinsp;\u0026plusmn;\u0026thinsp;1.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6.58\u0026thinsp;\u0026plusmn;\u0026thinsp;2.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.98\u0026thinsp;\u0026plusmn;\u0026thinsp;0.77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3.45\u0026thinsp;\u0026plusmn;\u0026thinsp;1.12\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eValues are expressed mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD. Values ​​shown with different letters indicate a significant difference between years according to the LSD test (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e\n\u003ch3\u003eRoot Length, Root Number, Root Development, Fresh Root Weight, and Dry Root Weight\u003c/h3\u003e\n\u003cp\u003eThe microbial fertilizer \u0026times; year interaction had a statistically significant effect on dry root weight, root length, and root development (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05), whereas its effect on fresh root weight and root number was not significant. While the treatments did not significantly influence fresh root weight, the year factor had a significant effect on this parameter (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.01). The treatments significantly affected root number (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.01), whereas the year factor did not show a significant influence on root number (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eEffect of applications on root development\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"10\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatments\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eFresh root weight (g) **\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003eDry root weight (g) *\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003eRoot length (cm) *\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003eRoot development\u003c/p\u003e \u003cp\u003e(0\u0026ndash;4 scale) **\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003eRoot number (piece)**\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e2024\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e2025\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e2024\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e2025\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e2024\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e2025\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e2024\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e2025\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cb\u003eCombined Mean\u003c/b\u003e\u003c/p\u003e \u003cp\u003e\u003cb\u003e(2024\u0026ndash;2025)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7.96\u0026thinsp;\u0026plusmn;\u0026thinsp;3.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.73\u0026thinsp;\u0026plusmn;\u0026thinsp;2.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.97\u0026thinsp;\u0026plusmn;\u0026thinsp;1.03 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.66\u0026thinsp;\u0026plusmn;\u0026thinsp;1.01 d-g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e14.0\u0026thinsp;\u0026plusmn;\u0026thinsp;2.00 b-d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e11.3\u0026thinsp;\u0026plusmn;\u0026thinsp;2.89 c-e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58 a-c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e15.67\u0026thinsp;\u0026plusmn;\u0026thinsp;2.88 bc\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-B\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.59\u0026thinsp;\u0026plusmn;\u0026thinsp;1.80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.36\u0026thinsp;\u0026plusmn;\u0026thinsp;0.65 e-g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.64\u0026thinsp;\u0026plusmn;\u0026thinsp;0.49 a-e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e11.3\u0026thinsp;\u0026plusmn;\u0026thinsp;5.51 c-e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e18.7\u0026thinsp;\u0026plusmn;\u0026thinsp;2.31 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58 a-d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e4.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e22.83\u0026thinsp;\u0026plusmn;\u0026thinsp;0.65 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e6.94\u0026thinsp;\u0026plusmn;\u0026thinsp;2.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.57\u0026thinsp;\u0026plusmn;\u0026thinsp;1.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.59\u0026thinsp;\u0026plusmn;\u0026thinsp;1.36 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.51 fg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e14.3\u0026thinsp;\u0026plusmn;\u0026thinsp;5.13 b-d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e10.3\u0026thinsp;\u0026plusmn;\u0026thinsp;2.08 c-e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00 a-d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e17.67\u0026thinsp;\u0026plusmn;\u0026thinsp;3.27 b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-D\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7.52\u0026thinsp;\u0026plusmn;\u0026thinsp;3.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.70\u0026thinsp;\u0026plusmn;\u0026thinsp;1.73 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.59\u0026thinsp;\u0026plusmn;\u0026thinsp;0.43 a-e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e15.7\u0026thinsp;\u0026plusmn;\u0026thinsp;4.04 a-c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e21.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00 a-d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 a-d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e16.33\u0026thinsp;\u0026plusmn;\u0026thinsp;3.39 bc\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-E\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e6.86\u0026thinsp;\u0026plusmn;\u0026thinsp;3.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.27\u0026thinsp;\u0026plusmn;\u0026thinsp;0.64\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.07\u0026thinsp;\u0026plusmn;\u0026thinsp;1.15 a-d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.97\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23 fg\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e13.7\u0026thinsp;\u0026plusmn;\u0026thinsp;3.79 b-d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e12.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00 c-e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2.7\u0026thinsp;\u0026plusmn;\u0026thinsp;1.15 b-d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58 a-c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e13.33\u0026thinsp;\u0026plusmn;\u0026thinsp;3.61 bc\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-F\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.87\u0026thinsp;\u0026plusmn;\u0026thinsp;1.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.80\u0026thinsp;\u0026plusmn;\u0026thinsp;0.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.26\u0026thinsp;\u0026plusmn;\u0026thinsp;1.08 b-f\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.37 g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e12.7\u0026thinsp;\u0026plusmn;\u0026thinsp;2.08 c-e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e13.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58 b-d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58 a-c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e12.67\u0026thinsp;\u0026plusmn;\u0026thinsp;2.73 c\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-G\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.59\u0026thinsp;\u0026plusmn;\u0026thinsp;1.73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57 a-d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.69\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28 c-g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e13.0\u0026thinsp;\u0026plusmn;\u0026thinsp;5.20 cd\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e14.3\u0026thinsp;\u0026plusmn;\u0026thinsp;1.15 b-d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58 c-e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58 a-c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e13.33\u0026thinsp;\u0026plusmn;\u0026thinsp;2.50 bc\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePositive control\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e6.02\u0026thinsp;\u0026plusmn;\u0026thinsp;1.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.47\u0026thinsp;\u0026plusmn;\u0026thinsp;1.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.84 a-c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.63 c-g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e7.3\u0026thinsp;\u0026plusmn;\u0026thinsp;1.15 e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e13.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00 cd\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e4.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e1.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58 e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e15.67\u0026thinsp;\u0026plusmn;\u0026thinsp;3.27 bc\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNegative control\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.33\u0026thinsp;\u0026plusmn;\u0026thinsp;1.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.70\u0026thinsp;\u0026plusmn;\u0026thinsp;1.06 a-e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.82\u0026thinsp;\u0026plusmn;\u0026thinsp;0.51 c-g\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e10.0\u0026thinsp;\u0026plusmn;\u0026thinsp;5.57 de\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e9.7\u0026thinsp;\u0026plusmn;\u0026thinsp;3.21 de\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58 a-c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e2.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00 de\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e12.17\u0026thinsp;\u0026plusmn;\u0026thinsp;3.54 c\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOverall mean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.96\u0026thinsp;\u0026plusmn;\u0026thinsp;2.46 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.84\u0026thinsp;\u0026plusmn;\u0026thinsp;1.74 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.95\u0026thinsp;\u0026plusmn;\u0026thinsp;1.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.69\u0026thinsp;\u0026plusmn;\u0026thinsp;0.77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e12.44\u0026thinsp;\u0026plusmn;\u0026thinsp;4.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e13.74\u0026thinsp;\u0026plusmn;\u0026thinsp;3.98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e2.96\u0026thinsp;\u0026plusmn;\u0026thinsp;0.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e15.52\u0026thinsp;\u0026plusmn;\u0026thinsp;4.73\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"10\"\u003eValues are expressed mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD. There is a significant difference between the values ​​indicated by different letters. ns: non-significant; *\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05; **\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.01\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe overall mean fresh root weight was found to be significantly higher in 2024 (5.96\u0026thinsp;\u0026plusmn;\u0026thinsp;2.46 g) compared to the overall mean in 2025 (3.84\u0026thinsp;\u0026plusmn;\u0026thinsp;1.74 g). The highest dry root weight was recorded in the MF-A treatment in 2024 (3.97\u0026thinsp;\u0026plusmn;\u0026thinsp;1.03 g), while the lowest value was obtained from the MF-F treatment in 2025 (0.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.37 g). The highest root length was recorded from the MF-D treatment in 2025 (21.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00 cm), whereas the lowest root length was obtained from the positive control in 2024 (7.3\u0026thinsp;\u0026plusmn;\u0026thinsp;1.15 cm). In terms of root development score, the positive control in 2024 (4.0) and the MF-B treatment in 2025 (4.0) exhibited the most favorable results, while the positive control in 2025 (1.3) showed the lowest root development level (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Regarding the effects of treatments on root number, the highest values were observed under MF-B (22.83\u0026thinsp;\u0026plusmn;\u0026thinsp;0.65 piece) and MF-C (17.67\u0026thinsp;\u0026plusmn;\u0026thinsp;3.27 piece), whereas the lowest root numbers were recorded in the MF-F treatment (12.67\u0026thinsp;\u0026plusmn;\u0026thinsp;2.73 piece) and the negative control (12.17\u0026thinsp;\u0026plusmn;\u0026thinsp;3.54 piece) (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eSapling Yield\u003c/h2\u003e \u003cp\u003eThe interaction between microbial fertilizer and year was found to be statistically significant for second-grade sapling yield (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05), whereas its effects on first size sapling yield and total sapling yield were not significant. In contrast, the effects of the treatments on both first size and total sapling yield were highly significant (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e; Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eEffect of microbial fertilizer applications on first size sapling yield, second size sapling yield and total sapling yield\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatments\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirst grade sapling yield (%) ***\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003eSecond grade sapling yield (%) *\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTotal sapling yield (%) ***\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eCombined Mean\u003c/b\u003e\u003c/p\u003e \u003cp\u003e\u003cb\u003e(2024\u0026ndash;2025)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e2024\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e2025\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003eCombined Mean\u003c/b\u003e\u003c/p\u003e \u003cp\u003e\u003cb\u003e(2024\u0026ndash;2025)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e46.0\u0026thinsp;\u0026plusmn;\u0026thinsp;3.18 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e11.7\u0026thinsp;\u0026plusmn;\u0026thinsp;3.34 d-f\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e12.3\u0026thinsp;\u0026plusmn;\u0026thinsp;2.15 c-f\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e57.9\u0026thinsp;\u0026plusmn;\u0026thinsp;3.61 d\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-B\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e57.0\u0026thinsp;\u0026plusmn;\u0026thinsp;7.05 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e13.3\u0026thinsp;\u0026plusmn;\u0026thinsp;6.11 c-e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e15.0\u0026thinsp;\u0026plusmn;\u0026thinsp;2.50 c-e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e71.2\u0026thinsp;\u0026plusmn;\u0026thinsp;5.16 ab\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e56.3\u0026thinsp;\u0026plusmn;\u0026thinsp;4.04 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e13.3\u0026thinsp;\u0026plusmn;\u0026thinsp;3.34 c-e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e12.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.50 c-f\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e69.2\u0026thinsp;\u0026plusmn;\u0026thinsp;4.77 b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-D\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e49.0\u0026thinsp;\u0026plusmn;\u0026thinsp;4.55 bc\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e10.0\u0026thinsp;\u0026plusmn;\u0026thinsp;3.33 ef\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e15.8\u0026thinsp;\u0026plusmn;\u0026thinsp;1.44 b-d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e61.9\u0026thinsp;\u0026plusmn;\u0026thinsp;3.28 cd\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-E\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e51.3\u0026thinsp;\u0026plusmn;\u0026thinsp;1.73 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e11.7\u0026thinsp;\u0026plusmn;\u0026thinsp;2.89 d-f\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e7.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.50 f\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e60.8\u0026thinsp;\u0026plusmn;\u0026thinsp;2.11 cd\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-F\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e47.0\u0026thinsp;\u0026plusmn;\u0026thinsp;3.40 bc\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e13.9\u0026thinsp;\u0026plusmn;\u0026thinsp;2.55 c-e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e21.0\u0026thinsp;\u0026plusmn;\u0026thinsp;3.61 ab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e63.9\u0026thinsp;\u0026plusmn;\u0026thinsp;5.96 c\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMF-G\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e40.7\u0026thinsp;\u0026plusmn;\u0026thinsp;2.91 d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e13.3\u0026thinsp;\u0026plusmn;\u0026thinsp;3.34 c-e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e22.5\u0026thinsp;\u0026plusmn;\u0026thinsp;4.33 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e58.6\u0026thinsp;\u0026plusmn;\u0026thinsp;6.51 d\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePositive control\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e57.8\u0026thinsp;\u0026plusmn;\u0026thinsp;6.23 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e17.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0,96 a-c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e15.3\u0026thinsp;\u0026plusmn;\u0026thinsp;2.52 c-e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e74.1\u0026thinsp;\u0026plusmn;\u0026thinsp;4.67 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNegative control\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e27.3\u0026thinsp;\u0026plusmn;\u0026thinsp;4.29 e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e15.6\u0026thinsp;\u0026plusmn;\u0026thinsp;5.09 cd\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00 c-e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e42.1\u0026thinsp;\u0026plusmn;\u0026thinsp;2.83 e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003eValues are expressed mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD. There is a significant difference between the values ​​indicated by different letters. ns: non-significant; *\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05; **\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.01; ***\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eWhen the effects of the treatments on first size sapling yield were evaluated, the highest first-grade yields were obtained from the positive control (57.8\u0026thinsp;\u0026plusmn;\u0026thinsp;6.23%), MF-B (57.0\u0026thinsp;\u0026plusmn;\u0026thinsp;7.05%), and MF-C (56.3\u0026thinsp;\u0026plusmn;\u0026thinsp;4.04%) treatments, whereas the lowest first-grade sapling yield was recorded in the negative control (27.3\u0026thinsp;\u0026plusmn;\u0026thinsp;4.29%). The highest total sapling yield was recorded in the positive control (74.1\u0026thinsp;\u0026plusmn;\u0026thinsp;4.67%), whereas the lowest total sapling yield was observed in the negative control (42.1\u0026thinsp;\u0026plusmn;\u0026thinsp;2.83%) (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eA correlation analysis was performed to evaluate the relationships among the parameters investigated. The strongest positive correlations were detected between fresh root weight and dry root weight (r\u0026thinsp;=\u0026thinsp;0.96), first-grade sapling yield and second-grade sapling yield (r\u0026thinsp;=\u0026thinsp;0.96), and fresh shoot weight and dry shoot weight (r\u0026thinsp;=\u0026thinsp;0.90). The highest negative correlation was found between shoot length and second-grade sapling yield (r\u0026thinsp;=\u0026thinsp;\u0026minus;\u0026thinsp;0.64) (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eAmong the notable positive correlations, strong relationships were observed between fresh shoot weight and shoot length (r\u0026thinsp;=\u0026thinsp;0.74), dry shoot weight and root development score (r\u0026thinsp;=\u0026thinsp;0.72), and dry shoot weight and root number (r\u0026thinsp;=\u0026thinsp;0.71). Moderate positive correlations were detected between first-grade sapling yield and root number (r\u0026thinsp;=\u0026thinsp;0.65), fresh shoot weight and root development score (r\u0026thinsp;=\u0026thinsp;0.63), and total sapling yield and root number (r\u0026thinsp;=\u0026thinsp;0.62). In contrast, moderate negative correlations were observed between fresh shoot weight and second-grade sapling yield (r\u0026thinsp;=\u0026thinsp;\u0026minus;\u0026thinsp;0.51), as well as between fresh root weight and second-grade sapling yield (r\u0026thinsp;=\u0026thinsp;\u0026minus;\u0026thinsp;0.43).\u003c/p\u003e \u003cp\u003eTo better elucidate the relationships between the treatments and the evaluated parameters, a Principal Component Analysis (PCA) was performed (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe first two principal components (Dim1 and Dim2) together accounted for 62.00% of the total variance. Total sapling yield, first-grade sapling yield, dry shoot weight, fresh shoot weight, shoot length, root development score, root number, and root length were positioned on the positive side of the first principal component axis (Dim1), which explained 40.4% of the total variance, whereas second-grade sapling yield was located on the negative side of this axis. Fresh shoot weight, root number, dry shoot weight, first-grade sapling yield, and root development score were found to be closely associated with the MF-B treatment. While total sapling yield was best represented by the MF-C treatment, shoot length was most strongly associated with the MF-A and MF-E treatments. Root length was related to the MF-B and MF-D treatments, whereas second-grade sapling yield was associated with the negative control and MF-G treatments. Fresh root weight and dry root weight were positioned on the positive side of the second principal component axis (Dim2), and both parameters were found to be closely associated with the MF-D treatment (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAccording to the hierarchical clustering heat map, the evaluated parameters were grouped into two main clusters (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e). The first main cluster consisted solely of second-grade sapling yield, while all other parameters were grouped within the second main cluster. This second main cluster was further divided into two sub-clusters. The first sub-cluster included root length, fresh root weight, and dry root weight. The second sub-cluster comprised root number, total sapling yield, first-grade sapling yield, shoot length, root development score, fresh shoot weight, and dry shoot weight. The heat map also classified the treatments into two principal groups. The first main treatment cluster was divided into two sub-clusters, with the first sub-cluster including MF-F, MF-G, and the positive control, and the second sub-cluster consisting of the negative control. The second main treatment cluster was also divided into two sub-clusters, with MF-D forming the first sub-cluster and MF-A, MF-B, MF-C, and MF-E constituting the second sub-cluster.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eIn the present study, the effects of microbial fertilizer applications on root and shoot development as well as sapling quality parameters were evaluated, and the results clearly demonstrated that microbial formulations exerted particularly pronounced effects on root morphology. The observed increases in root length, dry root weight, and root development score are consistent with previous reports indicating that arbuscular mycorrhizal fungi (AMF) and plant growth-promoting rhizobacteria (PGPR) enhance the root system through mechanisms such as phosphorus mobilization in the rhizosphere, stimulation of lateral root formation, and the production of hormone-like compounds (Azc\u0026oacute;n-Aguilar and Barea \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e1997\u003c/span\u003e; Ortiz-Castro et al. \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Cely et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). In particular, the fact that the MF-B, MF-C, and MF-D treatments exhibited performances comparable to the positive control strongly supports the notion that microbial inoculants play a powerful physiological and morphological stimulatory role in root tissues.\u003c/p\u003e \u003cp\u003eThe statistically significant \u0026ldquo;year \u0026times; treatment\u0026rdquo; interaction observed in this study further indicates that microbial efficacy is highly sensitive to environmental conditions. Previous studies have shown that soil moisture, temperature, pH, and nutrient availability are decisive factors regulating AMF activity and root colonization. It has been widely reported that the colonization rate and functional efficiency of AMF are strongly influenced by interannual environmental variability (Ortas \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Powell and Rillig \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Wahab et al. \u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Chen et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Conversely, some studies have suggested that AMF can provide relatively stable growth promotion regardless of year or environmental fluctuations (Bhardwaj et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Therefore, the year-dependent differences observed in root weight and root development level in the present study confirm the context-dependent and variable performance of microbial fertilizers.\u003c/p\u003e \u003cp\u003eThe limited and mostly non-significant effects of microbial treatments on shoot length and shoot biomass parameters are also in agreement with the existing literature. Numerous studies have reported that AMF influence aerial plant parts with lower sensitivity compared to the root zone (Geisen et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). However, in contrast, several investigations have shown that mycorrhizal applications can significantly enhance shoot growth under certain conditions (Wu et al. \u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Chandrasekaran \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Xu et al. \u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). These discrepancies suggest that microbial effectiveness is strongly dependent on factors such as plant species, inoculum density, root infection rate, and growing conditions.\u003c/p\u003e \u003cp\u003eIn this study, the significant effects of the \u0026ldquo;microbial fertilizer \u0026times; year\u0026rdquo; interaction on dry root weight, root length, and root development score (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) indicate that the response of grapevine roots to microbial applications is strongly modulated by environmental conditions and interannual variability. This finding is consistent with previous studies reporting that variations in soil moisture, temperature, and microbial activity across years can substantially influence root\u0026ndash;microorganism interactions (Torres et al. \u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Cardinale et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). In particular, the higher fresh root weight observed in 2024 (5.96\u0026thinsp;\u0026plusmn;\u0026thinsp;2.46 g) supports the findings of Lailheugue et al. (\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), who reported that environmental conditions play a decisive role in determining AMF-mediated root metabolism and colonization success.\u003c/p\u003e \u003cp\u003eThe highest dry root weight (3.97\u0026thinsp;\u0026plusmn;\u0026thinsp;1.03 g) was obtained from the MF-A treatment containing the \u003cem\u003eMethylobacterium symbioticum\u003c/em\u003e SB23 strain in 2024. This result agrees with previous findings demonstrating that \u003cem\u003eM. symbioticum\u003c/em\u003e produces indole-3-acetic acid (IAA), which stimulates lateral root formation and enhances root\u0026ndash;soil contact (Patten and Glick \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2002\u003c/span\u003e), thereby positively affecting root biomass (Bolla et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). In contrast, the low dry root weight observed under the MF-F treatment in 2025 suggests that the effects of microbial inoculants on root development may vary depending on plant genotype, inoculum characteristics, and seasonal conditions. Moreover, the highest root length recorded in 2025 under the MF-D treatment (21.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00 cm) may be attributed to differential microbial responsiveness of the rootstock used (Sportes et al. \u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eWith respect to root development score, the fact that both the positive control in 2024 and the MF-B treatment in 2025 achieved the highest score (4.0) indicates that microbial fertilizers can, under certain conditions, improve root morphology to a level comparable with that of natural microbial communities. However, the marked decline observed in the positive control in 2025 (1.3) highlights the pronounced role of the year factor in determining root development performance and is consistent with the literature emphasizing the significance of year-based variability in viticulture (Romero et al. \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2025\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn this study, root number was significantly affected by the treatments (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.01), whereas the effect of the year factor was not significant, indicating that microbial fertilizer applications can directly stimulate root branching independently of interannual variation. The highest root numbers obtained under the MF-B (22.83\u0026thinsp;\u0026plusmn;\u0026thinsp;0.65) and MF-C (17.67\u0026thinsp;\u0026plusmn;\u0026thinsp;3.27) treatments are in accordance with previous findings demonstrating that AMF and PGPR consortia enhance root branching (Trouvelot et al. \u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Torres et al. \u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Conversely, the low root number observed in the MF-F treatment suggests that not all microbial inoculant variants exert positive effects on root architecture and that root\u0026ndash;microorganism compatibility is a critical determinant of success.\u003c/p\u003e \u003cp\u003eOverall, these findings demonstrate that microbial fertilizer applications have a strong potential to enhance root architecture components such as root biomass, root length, root development score, and root number in Narince grapevine saplings; however, the magnitude of these effects varies depending on year conditions and application type. The literature reports that the sensitivity of grapevine root development to microbial inoculation is strongly influenced by environmental factors and the composition of the applied microbial populations (Goddard et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Lailheugue et al. \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Moukarzel et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), and the results of the present study are in close agreement with these observations.\u003c/p\u003e \u003cp\u003eIn general, microbial fertilizer applications exerted positive effects on root architecture and sapling quality class in grapevine nursery production, whereas the responses of shoot-related parameters remained limited and the overall effectiveness of the treatments varied between years. These results suggest that microbial fertilizers may be considered as an alternative or complementary biological input to chemical fertilizers under nursery conditions; however, achieving optimal efficacy requires careful selection of environmental conditions, microbial diversity, and application strategies.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis study comprehensively evaluated the effects of different microbial fertilizer formulations on container-grown grapevine sapling production in the Narince/110 R combination over a two-year experimental period. The findings revealed that microbial fertilizers exerted pronounced effects particularly on root morphology, sapling quality, and total sapling yield. The significant \u0026ldquo;microbial fertilizer \u0026times; year\u0026rdquo; interaction for root length, dry root weight, and root development score demonstrated that the performance of these applications is highly sensitive to environmental conditions. In contrast, the effects of the treatments on shoot length, fresh shoot weight, and dry shoot weight were generally limited.\u003c/p\u003e \u003cp\u003eThe MF-B, MF-C, and MF-D treatments emerged as the most effective in terms of root system performance and sapling quality; notably, the MF-B treatment exhibited superior performance with respect to root number, root development score, and first-grade sapling yield. Although the positive control (IBA) produced the highest total sapling yield in both years, the fact that several microbial fertilizer treatments (MF-B and MF-C) achieved results comparable to this application indicate that biological inputs may serve as an alternative or complementary option to chemical hormone-based treatments in nursery production.\u003c/p\u003e \u003cp\u003eCorrelation and PCA analyses confirmed that sapling yield is largely associated with root biomass and root functionality and clearly demonstrated that the primary mode of action of microbial fertilizers is mediated through the root system. This finding supports the potential of microbial fertilizers to improve sapling quality by enhancing root efficiency, particularly under limited volume growing media conditions.\u003c/p\u003e \u003cp\u003eOverall, this study demonstrates that microbial fertilizers represent a viable biological input for container-grown grapevine sapling production and can make substantial contributions to improving sapling yield and root system development. However, the variation in application performance between years highlights the need for further large-scale studies under different ecological regions, rootstock\u0026ndash;cultivar combinations and growing conditions. Such studies are essential for the standardization and widespread adoption of microbial fertilizers as a biological production input in viticulture.\u003c/p\u003e \u003cp\u003e \u003cb\u003eTablo 6\u003c/b\u003e Effects of Microbial Fertilizer Treatments and Control Groups\u003c/p\u003e\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"571\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eTreatment\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eComponent\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eMode of Action\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eConsistence with the Findings of the Present Study\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eMF-A\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cem\u003eMethylobacterium symbioense\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eEndophytic nitrogen fixation and phytohormone production\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eEnhanced shoot growth and increased fresh root weight\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eMF-B\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cem\u003eGlomus spp\u003c/em\u003e. + Humic acid, vitamins, and amino acids\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eRoot colonization, enhanced nutrient uptake, and hormone-like effects\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eHighest root number and sapling yield, with improved shoot growth\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eMF-C\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cem\u003eGlomus fasciculatum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003ePhosphorus uptake and root support\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eBalanced shoot and root development with good sapling yield\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eMF-D\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cem\u003eGlomus iranicum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eRoot elongation and enhanced stress tolerance\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eLongest root length, high root biomass, and good sapling yield\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eMF-E\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003ePGPR consortium (\u003cem\u003ePseudomonas\u003c/em\u003e, \u003cem\u003eBacillus\u003c/em\u003e, \u003cem\u003ePaenibacillus\u003c/em\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003ePhytohormone production and phosphate solubilization\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eHigh shoot development with low dry root weight\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eMF-F\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cem\u003eBacillus megaterium\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003ePhosphate solubilization and root stimulation\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eHigh proportion of second size saplings, with limited shoot and root development\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eMF-G\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cem\u003eTrichoderma harzianum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eBiocontrol and indirect growth promotion\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eModerate root development with a high proportion of second size saplings\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePositive control\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eSynthetic auxin (2000 ppm)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eRapid root stimulation\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eHigh root development score, but lower biomass and sapling yield compared to microbial fertilizers\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eNegative control\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eNo application\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003eLow values across all parameters\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors would like to thank Tokat Gaziosmanpaşa University, Faculty of Agriculture.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe research work was self-funded.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors and Affiliations\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDepartment of Horticulture, Faculty of Agricultural, University of Tokat Gaziosmanpaşa University, Turkey\u003c/p\u003e\n\u003cp\u003eAdem Yağcı\u003c/p\u003e\n\u003cp\u003eTokat Food Control Laboratory Directorate, Tokat, Turkey\u003c/p\u003e\n\u003cp\u003eİsa Hazar\u003c/p\u003e\n\u003cp\u003eDepartment of Horticulture, Faculty of Agriculture, Tokat Gaziosmanpaşa University, Turkey\u003c/p\u003e\n\u003cp\u003eRüstem Cangi\u003c/p\u003e\n\u003cp\u003eDepartment of Horticulture, Faculty of Agriculture, Kocaeli University, Turkey\u003c/p\u003e\n\u003cp\u003eSelda Daler\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors’ Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors contributed to the study conception and design. Material preparation, experimental conduction, data collection by Adem Yağcı, Rüstem Cangi, Selda Daler, İsa Hazar. The manuscript was written by İsa Hazar and all authors approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCorresponding author\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCorrespondence to\u0026nbsp;İsa Hazar\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate \u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable. This study did not involve human participants, human data, or animals.\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\u003eConflicts of Interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eİ. Hazar, A. Yağcı, S. Daler and R. Cangi declare that they have no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of Data and Material\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data generated during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAgnolucci M, Avio L, Pepe A, Turrini A, Cristani C, Bonini P, Cirino V, Colosimo F, Ruzzi M, Giovannetti M. Bacteria associated with a commercial mycorrhizal inoculum: Community composition and multifunctional activity as assessed by Illumina sequencing and culture-dependent tools. 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Plant Soil. 2001;235:235\u0026ndash;42. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1023/A:1011990013955\u003c/span\u003e\u003cspan address=\"10.1023/A:1011990013955\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Vitis vinifera L., Grafting, Seedling yield, Microbial inoculation, Root, Shoot","lastPublishedDoi":"10.21203/rs.3.rs-8475362/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8475362/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eIn Türkiye, grafted grapevine seedling production remains limited, and maintaining high seedling quality continues to be a major challenge. This study evaluated the effects of different microbial fertilizer (MF) formulations on seedling yield and quality in grafted grapevine production over two consecutive years (2024–2025). Scions of \u003cem\u003eVitis vinifera\u003c/em\u003e L. cv. Narince were grafted onto the 110 R rootstock and used as plant material. Seven microbial fertilizer treatments and two control groups (positive and negative) were tested. The microbial fertilizers contained either single microorganisms (\u003cem\u003eMethylobacterium symbioense, Glomus fasciculatum, Glomus iranicum, Bacillus megaterium,\u003c/em\u003e and \u003cem\u003eTrichoderma harzianum\u003c/em\u003e) or microbial consortia (\u003cem\u003ePseudomonas fluorescens, Paenibacillus polymyxa, Pantoea agglomerans,\u003c/em\u003e and \u003cem\u003eGlomus\u003c/em\u003e spp.). Treatments were applied to the basal ends of cuttings before planting. Seedling yield, shoot and root growht parameters were assessed two months after planting. Microbial fertilizers did not significantly affect shoot growth in the first year, whereas all treatments enhanced shoot growth in the second year compared with the controls. Root length, fresh root weight, and dry root weight increased significantly under all microbial fertilizer treatments. The highest total and first-grade seedling yields were obtained from the positive control (74.1% and 57.8%) and MF-B (71.2% and 57.0%) treatments. Strong positive correlations were observed between fresh and dry root weight (r = 0.96) and between first grade and total seedling yield (r = 0.96). Principal component analysis indicated that root morphology and seedling yield traits explained most of the total variation. These results demonstrate that microbial fertilizers can substantially improve seedling quality and production efficiency in grafted grapevine propagation by enhancing root system development.\u003c/p\u003e","manuscriptTitle":"Use of Microbial Fertilizers in Grapevine Seedling Production","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-01-06 13:52:59","doi":"10.21203/rs.3.rs-8475362/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"1ef55db2-bef7-4cb2-80cd-7753c6bf27f6","owner":[],"postedDate":"January 6th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-02-18T09:57:04+00:00","versionOfRecord":[],"versionCreatedAt":"2026-01-06 13:52:59","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8475362","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8475362","identity":"rs-8475362","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}