Efficacy Testing of Bio-fertilizer Based on Azotobacter on Sweet Corn (Zea mays convar)

Efficacy Testing of Bio-fertilizer Based on Azotobacter on Sweet Corn (Zea mays convar) | 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 Efficacy Testing of Bio-fertilizer Based on Azotobacter on Sweet Corn (Zea mays convar) Prateek Mishra, Surbhi Sharma, Shravan Kumar, Anjali Singh This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3854929/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 Excessive utilization of agrochemical fertilizers significantly leads to the decline of soil quality and its flora, it also affect human health. A material called bio-fertilizer contains living microorganisms that are advantageous to the growth and development of plants.Due to the impact of biofertilizers on enhancing crop yield, there is a growing focus on exploring beneficial microbes as biofertilizers within the agricultural domain. Consequently, this study was carried out to assess the efficacy of Azotobacterbiofertilizer on Sweet Corn ( Zea mays convar ).An experiment in a controlled artificial greenhouse setting involved 21 different treatments utilizing varying quantities of inoculants and carrier materials to create biofertilizer. The carrier material, derived from locally accessible raw materials such as cow dung, leaf litter, peat soil, lignite, charcoal, and vermiculite, serves as a substrate capable of hosting microbial inoculants and maintaining their viability for a specific duration, while also being easily accessible to farmers. Following the sowing of sweetcorn seeds for a month, the resulting germinated plants underwent analysis for parameters including plant height, stem girth, leaf count, length and width of leaves, as well as root length.The quantity of seed germination and the duration required for seeds to germinate were also recorded. Out of the 21 treatments, ST11, comprising 400 ml of inoculants combined with 300 g of carrier material, displayed higher values for plant parameters compared to the other treatments, although the statistical significance among all treatments was minimal. Each plant parameter responded differently to the various treatments. Contrary to the findings of others, the seeds immersed in microbial inoculant (ST2) did not notably enhance plant growth except for the number of leaves per plant. To validate the effectiveness of the formulated biofertilizer, a control was established: one set of seeds were sown solely in pot soil, while another contained only carrier material (ST21) without any microbial inoculant, which did not facilitate plant growth. Overall, plants treated with biofertilizer exhibited a positive response in terms of plant growth compared to the control group. Bio-fertilizer Soil fertility Azotobacter Zea mays convar Figures Figure 1 Figure 2 Introduction With the yearly rise in human population and the resulting increase in demand for food production and supply, a significant threat to food security is growing(Santhosh Kumar et al., 2018 ). Farmers have been using synthetic chemical fertilisers excessively to achieve high crop production in order to meet the rising food demand. However, excessive use of chemical fertilisers degrades soil quality by turning it excessively salty and biologically inert, which has detrimental effects on both the environment and human health. Additionally, it causes eutrophication, soil acidification, weakens plant roots, and an increase in disease occurrences. Due of all of these detrimental effects, employing chemical fertilisers has become into(Mma & Mfm, 2014 ). Given the adverse effects associated with chemical fertilizers, it has become crucial to adopt eco-friendly alternatives such as compost, farmyard manure, organic waste, and biofertilizers to boost crop productivity through environmentally sustainable methods(Singh, 2019 ).Bio-fertilizer, also recognized as "microbial inoculant," represents an innovative advancement in the field of agriculture.This organic formulation comprises live microorganisms extracted from soil or roots, which are then introduced into a compatible carrier substrate. This formulation aids in the conversion of essential plant nutrients in the soil, transforming them from an inaccessible form to a usable one for crop plants through biological processes(Boubekri et al., 2021 ).A carrier material is a base made from raw materials found locally, capable of holding microbial inoculants viable for a specific duration and easily accessible to farmers. It needs to be cost-effective, simple to handle, mixable, non-harmful to both the bacterial inoculants and plants, suitable for packaging, possessing high water retention capacity, and able to sustain the extended shelf life of microorganisms while being readily obtainable(Ben Rebah et al., 2002 ).Maize, often hailed as the superior cereal crop, is a demanding plant, needing approximately 120 to 180 kilograms of nitrogen per 10,000 square meters of land to thrive and produce a yield. Similarly, Zea mays convar , belonging to the maize species, has a substantial need for nitrogen fertilizer to support its growth and productivity. These plants thrive best in well-drained, deep loam, and silt loamy soils rich in organic matter, with an ideal pH level ranging from 7.5 to 8.5. They exhibit optimal growth at temperatures around 34°C.Sweetcorn typically requires approximately 90 days to complete its growth and maturation process. Reports indicate that biofertilizers have the capacity to fix approximately 20–40 kilograms of nitrogen per 4046 square meters of land(Mahato & Kafle, 2018 ). Within the realm of biofertilizers, nitrogen-fixing bacteria like Azotobacter are recognized for their ability to fix nitrogen as ammonium ions in the soil(Agarwal et al., 2018 ). Additionally, they generate plant growth hormones such as indole acetic acid (IAA), gibberellins, cytokinin, and auxin, contributing to processes such as plant cell division, flowering, seed germination, seed dormancy, abscission, and offering protection against pathogens(Potdar et al., 2019 ). Numerous research studies have been conducted regarding the implementation of biofertilizers on crop plants, resulting in a noteworthy enhancement in vegetative growth, yield, and soil fertility. The application of Azotobacter has shown significant positive effects on the growth and yield of maize, leading to increased plant height, root length, stem diameter, fresh and dry weight of maize, leaf area, number of kernels per row, grain weight, and maize stover yield(Baral& Adhikari, 2014 ). Moreover, the utilization of biofertilizers has demonstrated improvements in grain yield, stimulation of seed germination, bolstering seedling resistance against both biotic and abiotic stress, nitrogen fixation, and the production of phytohormones, thereby promoting better growth in maize plants(Mirjana Jarak, 2012 ).Rudolph et al., ( 2015 ) findings revealed that the application of plant growth promoting rhizobacteria on maize seeds notably elevated seed germination, biomass, and overall yield (LG, 2015 ). Similarly,(Lhamo et al., 2022 ) observed that treating maize seeds with Azotobacter resulted in higher chlorophyll content within the leaves. This biofertilizer also notably improved the growth of Zea mays, thereby advocating its use, especially for enhancing maize production(Mirjana Jarak, 2012 ).Potdar et al. ( 2019 ) discovered that applying Azotobacter derived from lignite to sweet corn seeds resulted in heightened growth and increased yield of sweet corn. In a comparative study conducted by Mahato & Neupane ( 2018 ) between Azotobacter and Trichoderma, along with other fertilizers, on maize growth, seeds treated with Azotobacter displayed significant improvements in parameters such as plant height, stem girth, dry shoot weight, root length and width, as well as root weight. Meanwhile, the application of Trichoderma exhibited either negative effects or negligible impact. Similar findings were also noted in two cultivars of Zea mays L., namely HUDAIBA (HD) and MUGTAMA45 (MG).Hence, the current research endeavors to isolate Azotobacter, create biofertilizer utilizing Azotobacter, and conduct trial experiments to assess its effectiveness as a biofertilizer for sweetcorn ( Zea mays convar ) cultivation. Material and methods 2.1 Isolation of microorganism The soil sample was collected from pee field within the CSA Campus. Soil samples were obtained from the rhizosphere area around Pea plants. These soil samples, enclosed in sterile plastic bags, were transported to the laboratory to isolate Azotobacter. A stock solution was prepared by combining 1 gram of soil sample with 9 milliliters of distilled water in a test tube. Subsequently, a series of dilutions were conducted: initially, a 10-fold dilution (10 − 1) was achieved by adding 1 milliliter of the stock solution to 9 milliliters of distilled water using a sterilized pipette. This process was repeated for subsequent dilutions, such as 10 − 2, 10 − 3, and so on, until reaching a 10 − 9 dilution factor. The goal of these serial dilutions in this experiment was to decrease the density of cell cultures to a more manageable concentration, ensuring that the colonies formed would be distinct and less dense.Following the serial dilution process, a specialized nitrogen-free Ashby mannitol media was prepared to isolate Azotobacter. Compostion of Ashby mannitol media are discussed in Table 4 . Subsequently, 100 microliters (0.1 ml) of diluted samples from 10 − 3, 10 − 5, 10 − 7, and 10 − 9 were transferred from the test tubes to the Ashby culture media using a sterilized pipette. These specific dilution factors were selected to decrease bacterial concentration, ensuring that the cultures grown in the plate would not be dense, facilitating the growth of distinct, pure colonies of the microbes. The culture was evenly spread on Ashby media using a spreader and then incubated at 30˚C for 7 days. After incubation, pure cultures were isolated by streaking the colonies grown on the culture plate onto a new culture plate containing Ashby media, followed by another incubation for 3 days at 30˚C. All procedures were conducted under Laminar Air Flow (LAF) to maintain aseptic conditions. The streak plate technique was utilized to separate colonies of pure Azotobacter culture from the mixed microbial population, effectively thinning out bacterial cells to ensure they were well-separated and adequately spaced. 2.2 Characterization Once bacterial strains were cultured in a pure environment, a small amount of the strain was utilized for gram staining. Following this, a motility test was carried out using the hanging drop method to assess the bacteria's motile properties. Additionally, biochemical tests including the methyl red test, VP test, starch hydrolysis, and catalase test were conducted to validate the identity of the bacterial strain. 2.3 Carrier Material The components used as a carrier material for creating biofertilizer were derived from readily accessible local resources, including peat soil (P), forest organic matter obtained from leaf litter (F), and cow dung (C). These materials were gathered, dried under the sun, ground, sieved, and combined in a ratio of 1 part peat soil to 2 parts leaf litter to 2 parts cow dung, resulting in a final mixture of 4 kilograms. Subsequently, the carrier material underwent sterilization in an autoclave at 121°C for 15 minutes. 2.4 Inoculation To prepare the microbial inoculants for mixing with the carrier material, a bacterial culture was cultivated in broth by transferring bacterial colonies into 500 milliliters of Azotobacter broth. This broth was then incubated in a shaking incubator at 30˚C, with an agitation speed of 200 revolutions per minute (rpm) for 6 days to foster mass culture. A total volume of 5000 milliliters of Azotobacter broth was generated. The biofertilizer formulation utilized the broth culture once the cell count of the inoculum reached 10 8 − 10 9 cells/ml. However, in this particular investigation, the growth rate of Azotobacter in the liquid medium (Azotobacter broth) was assessed by measuring the optical density using a spectrophotometer at a wavelength of 550 nm.When the optical density reached 1.334, the inoculum was combined with the carrier material. Different ratios were considered for mixing the carrier material with the broth culture: for every 1 kilogram of carrier material, sterile containers were used to mix 100 milliliters, 250 milliliters, and 400 milliliters of broth culture, respectively. Following thorough mixing, the carrier material containing the inoculant was allowed to cure for 6 days in a cool, dry place, kept away from direct sunlight and heat. Following the curing period of six days, one gram from each combination was extracted to isolate the microbe, aiming to assess the bacterial density and confirm the presence of the microbial organism in the carrier material. The bacterial density was gauged using a spectrophotometer, revealing optical densities (OD) of 1.3, 1.4, and 1.5 at 550 nm for carrier materials treated with 100 ml, 250 ml, and 400 ml of the broth culture, respectively. The culture obtained underwent a biochemical test, confirming the presence of Azotobacter as the test yielded positive results for this specific microorganism. 2.5 Soil Preparation An initial trial was carried out using the prepared biofertilizer in a plastic container within a greenhouse setting. Soil samples were gathered from the garden area at HBTU Campus, Kanpur. These soil samples were subjected to air-drying, grinding, sieving (2 mm), and thorough mixing. The botany laboratory at Sherubtse College conducted qualitative assessments of soil parameters, including soil organic carbon, organic matter, porosity, water holding capacity, soil texture, soil conductivity, and pH. These analyses were conducted both before and after the application of the biofertilizer to assess its impact on soil properties. 2.6 Experimental Design All the procedures were conducted within a greenhouse environment using plastic pots situated near the botany laboratory at Chandrashekar Azad University, spanning from February to April 2023. The experimental setup encompassed 21 treatments arranged in a randomized block design, with three replicates for each treatment, and each pot represented a single replicate, totaling 63 pots. Each plastic pot, measuring 30x28 cm, was filled with approximately 5 kilograms of thoroughly mixed soil. Following soil filling, a recommended quantity of biofertilizer was added and mixed into the soil within the pots. Subsequently, seeds were sown. Additionally, a control experiment was established wherein seeds were sown in potted soil without the incorporation of any formulated biofertilizer, aimed at evaluating the efficacy of the biofertilizer. When required, the pots were watered with 250 milliliters of tap water. The details outlining the 21 different treatments are presented in Table 1 . Table 1 Different types of treatment provided to the soil Soil Treatments Replica Amount of Biofertilizer Untreated Soil ST1 Control Seed Treatment ST2 Seed treated ST3 100 ml, 100 g Soil was subjected ST4 100 ml, 200 g to Biofertilizer ST5 100 ml, 300 g treatment before ST6 250 ml, 100 g planting the seeds ST7 250 ml, 200 g ST8 250 ml, 300 g ST9 400 ml, 100 g ST10 400 ml, 200 g ST11 400 ml, 300 g Soil treatment was done two times: First, before top Dressing after two ST12 ST13 ST14 ST15 100ml, 100g 100ml, 200g 100ml, 300g 250ml, 100g Weeks of seed ST16 250 ml, 200 g planting ST17 250 ml, 300 g ST18 400 ml, 100 g ST19 400 ml, 200 g ST20 400 ml, 300 g After addition of carrier matreial ST21 No inoculants 2.7 Data collection and analysis After a month of sowing, data was collected on the height of the plants, the number of leaves on each plant, the length and width of the leaves, the stem girth, the number and length of the roots. The highest point of the uppermost leaf's arch, where the tip points downward, was used to measure the height of the plant. Leaf width was measured at the point where the leaf reached its maximum width, and leaf length was measured from the tip of the leaf to the point where it joins the leaf sheath. There was also a count of the leaves on each plant. Both the primary root's length and the total number of roots were measured. Using a ruler, the plant's height, width, length of leaves, and length of roots were measured in centimetres. A millimeter-based vernier calliper was used to measure the stem's girth.Graphpad Prism was utilised to analyse the replicated data. In a randomised complete block design, this was utilised to compute analysis of variance (ANOVA), standard errors, variance, and coefficient of variation. To determine if there were any differences amongst the 21 soil treatments, ANOVA was performed. To compare all potential pairwise treatments, Tukey's Multiple Range Test was used. A 5% level of significance (P < 0.05) was used to estimate the significance of the differences between the treatment combinations. Result and discussion 3.1 Isolation of microorganism Using Ashby mannitol medium, a selective medium for Azotobacter, the nitrogen-fixing bacteria was successfully isolated, and their growth in the culture media confirms Azotobacter. Both morphological and biochemical tests were used to characterise the isolated bacteria. The colonies were mucoid, white, milky, round, uneven, and soft. The goal of identifying bacteria is to verify that the strain that was grown or obtained is the desired microbe. Isolating bacteria is necessary to obtain desired microbes for the study. The colonies were counted using a standard plate count method after they had formed on the culture media. 3.2 Characterization Gramme staining was used to verify that Azotobacter was present. These bacteria are gram-negative and have a safranin red colour. Azotobacter has an oval, rod-shaped pleomorphism. Since Azotobacter is a motile bacterium, the bacterial motility was tested using the hanging drop method. It was discovered that the material under the microscope was motile because rod- and oval-shaped bacteria could be seen moving in a particular kind of wet mount. Through biochemical characterization of Azotobacter, the isolated strain was verified; starch hydrolysis, methyl red, Voges-Proskauer (VP), and the catalase test all yielded positive results. The biochemical test result for Azotobacter is displayed in Fig. 1 . 3.3 Analysis of soil Prior to and following the treatment, the texture, organic carbon, organic matter, porosity, pH, and soil conductivity of the soil used in the pot experiment were examined. Sweetcorn is recognised to thrive in loam soil that has a pH ranging from 5.8 to 7 when organic matter is properly supplied. The data collected for soil properties both before and after treatments are displayed in Fig. 2 . The results are consistent with the research of Mahato&Neupane (2017), who found that applying biofertilizer enhances the physical characteristics of the soil and increases soil fertility and nutrient availability to plants. Additionally, organic matter increases the ability of microbes to hold onto water and gives them nutrients, which promotes microbial activity. According to Umesha et al. (2014), organic matter also increases the soil's ability to hold water and gives microbes nutrients, which promotes microbial activity and supports plant growth. Additionally, it was stated that biofertilizers contributed to the fertility of the soil. Usingbiofertilizer increases the amount of organic matter in the soil, which is a crucial sign of soil fertility. 3.4 Measurement of development plant Following a month of sowing, the data was recorded regarding various growth parameters. Table 2 illustrates how the mean values are arranged in ascending order of the values.Contrary to some previous work done, this study showed that the plant height, stem girth, leaf length, and root length were the lowest. This could be because the seeds were treated differently with Azotobacter inoculants. In this study, seeds were kept in the shade for 30 minutes prior to sowing after being soaked in Azotobacter broth for 20 minutes without the use of any sticking agents. Table 2 Imapct of biofertilizer on development of sweetcorn. Soil Treatment Plant height (cm) Stem girth(mm) No. of leaves Leaf length (cm) Leaf width (cm) Root length(cm) No. of seed germinated Days taken to germinate ST1 21.7 2.0 3.7 13.8 1.8 11.7 3.6 13.0 ST2 19.7 2.8 5.5 12.3 1.9 11.0 3.6 11.0 ST3 21.5 3.2 3.7 14.6 1.8 11.3 4.6 11.0 ST4 24.7 3.2 3.9 18.2 1.9 13.5 5.3 11.0 5T5 25.3 3.9 3.6 17.8 1.9 11.0 5.0 9.6 ST6 24.8 2.9 3.6 18.7 2.2 13.8 5.0 11.0 ST7 24.5 2.9 3.8 18.3 1.9 12.0 5.0 9.4 ST8 26.7 2.8 3.8 18.2 1.8 12.9 5.3 9.0 ST9 26.9 3.0 4.0 17.9 1.8 14.3 5.0 10. ST10 26.6 2.9 3.9 19.1 1.9 13.5 5.6 9.0 ST11 28.5 3.0 3.8 19.5 1.9 12.7 5.6 9.0 ST12 21.6 2.6 3.7 14.6 1.8 12. 5.3 11.0 ST13 23.1 2.9 3.6 16.6 1.9 12.4 6.3 11.0 ST14 23.2 2.6 3.6 15.7 1.8 12.4 4.6 9.0 ST15 23.7 2.7 3.7 16.1 1.8 13.3 6.0 9.6 T16 23.4 3.0 3.7 16.9 1.9 11.4 6.0 9.6 ST17 23.9 2.8 3.8 16.7 1.8 11.8 5.6 9.0 ST18 24.8 2.8 3.8 17.1 1.9 12.9 5.6 9.0 ST19 25.3 2.6 3.6 17.5 1.9 12.4 6.0 9.0 ST20 25.5 2.6 3.6 18.9 1.8 13.0 5.3 9.0 ST21 22.6 2.5 3.6 16.5 1.7 12.0 3.6 11.6 The results of this study indicate that the application of biofertilizers did not significantly (P < 0.05) affect the vegetative growth parameters. Out of all the treatments, soil treatment 11 (ST11) had the tallest plant (28.5 cm) and leaf length (19.5 cm). Soil treatment 2 (ST2) had the lowest plant height, indicating a detrimental effect over the control setup. ST2 yielded the highest number of leaves ever recorded. ST6 had the largest leaf width (2.2 cm), while ST21 had the smallest leaf width (1.7 cm). This led to an adverse effect on the control setup. According to similar research, leaf length was marginally greater in soil-inoculated Azotobacterbiofertilizerand lowest in seed-inoculated Azotobacter. In addition, Khushali et al. (2015) found that, in comparison to other treatments, leaves treated with Azotobacter seeds were smaller. Soil treatment 5 (ST5) recorded the maximum stem girth (3.2 mm), soil treatment 6 (ST6) the leaf width (2.2 cm), soil treatment 9 (ST9) the root length (14.3 cm), and soil treatment 15 (ST15), 16 (ST16), and 19 (ST19) the maximum number of seeds that germinated. As shown in Table 1 , biofertilizer is applied to each treatment method in varying quantities. Some previous works by various authors has stated the increased rate of organic fertiliser is what caused the increase in plant parameters, which may be explained by the fact that it contains a good amount of nutrients both (micro and macro nutrients) that is essential for the growth and development of plants.In addition, Mirza et al. (2000) reported that biofertilizers facilitate the synthesis of substances that promote plant growth, such as auxin, which raises lethal fibres and enhances plant growth in addition to fixing nitrogen. The lack of significant variation in plant parameters between those treatments may be due to the plant parameters' high genetic influence, which has no discernible impact on the application of biofertilizers. The application of biofertilizer had a significant impact on parameters such as the number of days taken for germination. The number of seeds that germinated varied amongst treatments, but there was no statistically significant difference; however, ST15, ST16, and ST19 showed the highest levels of seed germination. The maximum number of days required for germination in ST1 was 13, and the lowest number of days required was 8 in soil treated with biofertilizer (ST8, ST10, ST11, ST14, and ST17–T20). However, the number of days required for germination following the application of biofertilizer was statistically significant with the control setup (ST1). The outcome was consistent with the previous researches, that shown using biofertilizer increased seed germination. 3.5 A comparative analysis of soil treated once (ST3–ST11) twice and (ST12–ST20): The means in the same column that are followed by the same letters do not differ significantly from one another. The means from ST3– ST11 are represented by ST-1, and the means from T12–T20 are represented by ST-2.Nine of the twenty-one treatments (ST3–ST11) were single treatments, wshile the other nine treatments (ST12–ST20) are double treated, meaning that the soil is treated with biofertilizers twice: once during seed sowing and again two weeks later when the plants receive a topdressing of biofertilizers. Since maize is a heavy feeder crop that needs a lot more nutrients than other crops, it received two treatments. Table 3 presents the mean values for soil treated with single treatment (ST3-ST11) and double treatment (ST12- ST20). There was a statistically significant difference in plant height, stem girth, and leaf length between the treatments. Other plant characteristics, however, including the quantity of leaves and roots, the width and length of the leaves, the amount of chlorophyll, the number of seeds that germinated, and the time it took for those seeds to germinate, did not differ significantly between the two treatments. Table 3 comparative analysis of soil treated once (ST3- ST11) and twice (ST12-ST20) Soil Treatment Plant height (cm) Stem girth(mm) No. of leaves Leaf length (cm) Leaf width (cm) Root length(cm) No. of seed germinated Days taken to germinate ST-1 25.5 3.06 3.789 18.03 1.8 12.77 5.1 9.88 ST-2 23.8 273 3,677 16.64 1.84 12.44 5.6 9.57 However, compared to double treatment, single treated soil produced higher plant growth values. This could be because the plants were uprooted for analysis after two weeks of receiving biofertilizer topdressing. It's possible that the microbial activity and nutrients needed to support plant growth weren't completed in the allotted two weeks. Additionally, the amount, timing, and technique of topdressing may have varied from other methods because Mahato & Neupane (2017) used the dibbling method to topdress fertiliser 26 days after sowing. Others suggested that top dressing be applied when the maize plant reaches a height of approximately 18 inches, or six to eight weeks following the plant's emergence which gives effect of topdressing. Nevertheless, topdressing was carried out in the current study following two weeks of seeding. Table 4 List of Chemical Used In Ashby Mannitol Media Ingredients (gm per 100 ml) composition Mannitol 2gm Di potassium phosphate 0.02gm Magnesium sulphate 0.02gm Sodium chloride 0.02gm Potassium sulphate 0.01gm Calcium carbonate 0.5gm Agar- agar 1.5gm Conclusion This study contributes to the growing body of knowledge on importance of biofertilizers as an environmentally friendly substitute for conventional fertilisers. It was clear from a thorough examination of numerous plant growth characteristics that the sweetcorn plants benefited from the application of biofertilizer. According to the results, biofertilizer may be able to raise sweetcorn crops' overall productivity. Additionally, the statistical analyses offered a strong basis for determining the importance of variations among different treatments. These analyses strengthened the validity of the observed results by giving the study a quantitative component. As we move forward, it is crucial to acknowledge the potential implications of incorporating biofertilizer into sweetcorn cultivation practices. Nonetheless, the samples treated with the specially blended biofertilizer had fewer days for the seeds to germinate. So, more research focusing on measuring the yield components is required to comprehend the efficacy of various treatments on Zea mays convar . The favourable results reported in this study provide opportunities for additional investigation and use of biofertilizer in various agricultural contexts. Declarations 8.1 Ethical Approval Not applicable 8.2 Consent to Participate All participants involved in this study provided informed consent before participating, understanding the purpose and procedures involved. 8.3 Consent to Publish I confirm that all co-authors have reviewed and given their consent for the publication of this research paper. 8.4 Authors Contributions We would like to express sincere gratitude to Shravan kumar for their invaluable guidance and support in shaping manuscript to be published. Prateek Mishrahas contributed significantly to the conception, design and writing of the research. Anjali Singh has contributed in data collection and lab work and Surbhi Sharma has contributed in the data analysis, and interpretation of the research. 8.5 Funding Not applicable 8.6 Competing Interests: The authors declare no competing interests that could influence the objectivity, integrity, or validity of the research. References Agarwal P, Gupta R, Gill IK (2018) Importance of biofertilizers in agriculture biotechnology. Biologigal Res 9(3):1–13 www.scholarsresearchlibrary.com Aloo BN, Makumba BA, Mbega ER, Shilev S (2022) Plant growth-promoting rhizobacterial biofertilizers for crop production: The past, present, and future . September , 1–15. https://doi.org/10.3389/fpls.2022.1002448 Baral BR, Adhikari P (2014) Effect of Azotobacter on Growth and Yield of Maize. 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Adv Plants Agric Res 2(6):250–253. https://doi.org/10.15406/apar.2015.02.00069 Lhamo T, Bajgai RC, Gurung B (2022) Efficacy of Azotobacter Biofertilizer for Popcorn (Zea mays var. everta (Sturtev.) Zhuk.) . 15 , 24–36 Mahato S, Kafle A (2018) Comparative study of Azotobacter with or without other fertilizers on growth and yield of wheat in Western hills of Nepal. Annals Agrarian Sci 16(3):250–256. https://doi.org/10.1016/j.aasci.2018.04.004 Mahato S, Neupane S (2018) Comparative study of impact of Azotobacter and Trichoderma with other fertilizers on maize growth. J Maize Res Dev 3(1):1–16. https://doi.org/10.3126/jmrd.v3i1.18915 Mirjana Jarak (2012) Effects of plant growth promoting rhizobacteria on maize in greenhouse and field trial. Afr J Microbiol Res 6(27). https://doi.org/10.5897/ajmr12.759 Mma Y, Mfm E (2014) Biofertilizers and their role in management of plant parasitic nematodes. A review. 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Seed Sci Technol 43(3):507–518. https://doi.org/10.15258/sst.2015.43.3.04 Santhosh Kumar M, Chandramohan Reddy G, Phogat M, Korav S, Santhosh Kumar CM (2018) Role of bio-fertilizers towards sustainable agricultural development: A review. ~ 1915 ~ J Pharmacognosy Phytochemistry 7(6):1915–1921 Shahwar D, Mushtaq Z, Mushtaq H, Alqarawi AA (2023) Heliyon Role of microbial inoculants as bio fertilizers for improving crop productivity: A review. Heliyon 9(6):e16134. https://doi.org/10.1016/j.heliyon.2023.e16134 Singh I (2019) Microbial Biofertilizers: Types and Applications Metadata of the chapter that will be visualized online. Nat Biofertil Sustanainability Agricultural. August. https://doi.org/10.1007/978-3-030-18933-4 Suhag M (2016) Potential of Biofertilizers to Replace Chemical Fertilizers . April , 1–6. https://doi.org/10.17148/IARJSET.2016.3534 Sumbul A, Ansari RA, Rizvi R, Mahmood I (2020) Saudi Journal of Biological Sciences Azotobacter: A potential bio-fertilizer for soil and plant health management. Saudi J Biol Sci 27(12):3634–3640. https://doi.org/10.1016/j.sjbs.2020.08.004 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-3854929","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":276104970,"identity":"b08ef939-d11d-46fc-badd-26c1917a23e5","order_by":0,"name":"Prateek Mishra","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABJ0lEQVRIiWNgGAWjYDACZgY2OJuxoYKtnrG9Acg0sCBCCxtIyxm+BOaeAyAtEvjsQdLS2CaXwD4jAcTFrUW3nf3Z44qaO/n885sPPpzZZpbHO/P51Q0/CiQY+Nu7E7BpMTvMY2545tgzyxnH2JINN5xLK5acnVN2swfoMIkzZzfg0MIm2cB22IDhGI+Z5IOyY4wbZ+ek3eABajGQyMWhhf2ZZMO/wwbyx/i//3zA9p9x/80zaTf/4NXCYCbZ2HbYwOAYDxvjhja2xMYZ7Mdu47cF6J7GvmcGhsfSjCVnnGEzZuzJYbstYyDBg9Mv548DHfbtjoHc4cMPP/ZUsMkxth9/dvPNHxs5/vZerFqg4AAyh8cATOJRjqGF/QEB1aNgFIyCUTDCAAABvWoYPtHajwAAAABJRU5ErkJggg==","orcid":"","institution":"Harcourt Butler Technical University","correspondingAuthor":true,"prefix":"","firstName":"Prateek","middleName":"","lastName":"Mishra","suffix":""},{"id":276104971,"identity":"8a56eb6c-7b97-4b17-9026-b72a6d53fa99","order_by":1,"name":"Surbhi Sharma","email":"","orcid":"","institution":"Harcourt Butler Technical University","correspondingAuthor":false,"prefix":"","firstName":"Surbhi","middleName":"","lastName":"Sharma","suffix":""},{"id":276104972,"identity":"6b6032fc-a4e7-4ed1-9750-e81351a24977","order_by":2,"name":"Shravan Kumar","email":"","orcid":"","institution":"Harcourt Butler Technical University","correspondingAuthor":false,"prefix":"","firstName":"Shravan","middleName":"","lastName":"Kumar","suffix":""},{"id":276104973,"identity":"e708e980-2606-407c-b822-fed28f07e101","order_by":3,"name":"Anjali Singh","email":"","orcid":"","institution":"CSIR-Indian Institute of Toxicology Research","correspondingAuthor":false,"prefix":"","firstName":"Anjali","middleName":"","lastName":"Singh","suffix":""}],"badges":[],"createdAt":"2024-01-11 22:14:09","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3854929/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3854929/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":52074478,"identity":"b5effe5e-fd4c-4fd7-9ec5-92d45ac5ecb9","added_by":"auto","created_at":"2024-03-06 09:11:24","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":68935,"visible":true,"origin":"","legend":"\u003cp\u003eA. Microscopic view of Azotobacter, B. Bacterial culture, C. Positive test for starch hydrolysis test, D. Bubble formation on catalase test, E-F. Positive test for MR-VP test.\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3854929/v1/023958efe82d05dcb712b62b.jpg"},{"id":52074779,"identity":"5926f667-b9db-49ef-b1dd-cb0d994cd7fe","added_by":"auto","created_at":"2024-03-06 09:19:24","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":103851,"visible":true,"origin":"","legend":"\u003cp\u003eBefore and after treatment comparison of soil properties\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-3854929/v1/80c61d2a211ed251cf532026.png"},{"id":52930544,"identity":"003eadd8-4e00-4c10-87e9-5a085830991f","added_by":"auto","created_at":"2024-03-18 20:11:06","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":523213,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3854929/v1/3da94d61-ab70-436e-a111-f8aa31e1364f.pdf"}],"financialInterests":"","formattedTitle":"Efficacy Testing of Bio-fertilizer Based on Azotobacter on Sweet Corn (Zea mays convar)","fulltext":[{"header":"Introduction","content":"\u003cp\u003eWith the yearly rise in human population and the resulting increase in demand for food production and supply, a significant threat to food security is growing(Santhosh Kumar et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Farmers have been using synthetic chemical fertilisers excessively to achieve high crop production in order to meet the rising food demand. However, excessive use of chemical fertilisers degrades soil quality by turning it excessively salty and biologically inert, which has detrimental effects on both the environment and human health. Additionally, it causes eutrophication, soil acidification, weakens plant roots, and an increase in disease occurrences. Due of all of these detrimental effects, employing chemical fertilisers has become into(Mma \u0026amp; Mfm, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Given the adverse effects associated with chemical fertilizers, it has become crucial to adopt eco-friendly alternatives such as compost, farmyard manure, organic waste, and biofertilizers to boost crop productivity through environmentally sustainable methods(Singh, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2019\u003c/span\u003e).Bio-fertilizer, also recognized as \"microbial inoculant,\" represents an innovative advancement in the field of agriculture.This organic formulation comprises live microorganisms extracted from soil or roots, which are then introduced into a compatible carrier substrate. This formulation aids in the conversion of essential plant nutrients in the soil, transforming them from an inaccessible form to a usable one for crop plants through biological processes(Boubekri et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).A carrier material is a base made from raw materials found locally, capable of holding microbial inoculants viable for a specific duration and easily accessible to farmers. It needs to be cost-effective, simple to handle, mixable, non-harmful to both the bacterial inoculants and plants, suitable for packaging, possessing high water retention capacity, and able to sustain the extended shelf life of microorganisms while being readily obtainable(Ben Rebah et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2002\u003c/span\u003e).Maize, often hailed as the superior cereal crop, is a demanding plant, needing approximately 120 to 180 kilograms of nitrogen per 10,000 square meters of land to thrive and produce a yield. Similarly, \u003cem\u003eZea mays convar\u003c/em\u003e, belonging to the maize species, has a substantial need for nitrogen fertilizer to support its growth and productivity. These plants thrive best in well-drained, deep loam, and silt loamy soils rich in organic matter, with an ideal pH level ranging from 7.5 to 8.5. They exhibit optimal growth at temperatures around 34\u0026deg;C.Sweetcorn typically requires approximately 90 days to complete its growth and maturation process.\u003c/p\u003e \u003cp\u003eReports indicate that biofertilizers have the capacity to fix approximately 20\u0026ndash;40 kilograms of nitrogen per 4046 square meters of land(Mahato \u0026amp; Kafle, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Within the realm of biofertilizers, nitrogen-fixing bacteria like Azotobacter are recognized for their ability to fix nitrogen as ammonium ions in the soil(Agarwal et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Additionally, they generate plant growth hormones such as indole acetic acid (IAA), gibberellins, cytokinin, and auxin, contributing to processes such as plant cell division, flowering, seed germination, seed dormancy, abscission, and offering protection against pathogens(Potdar et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eNumerous research studies have been conducted regarding the implementation of biofertilizers on crop plants, resulting in a noteworthy enhancement in vegetative growth, yield, and soil fertility. The application of Azotobacter has shown significant positive effects on the growth and yield of maize, leading to increased plant height, root length, stem diameter, fresh and dry weight of maize, leaf area, number of kernels per row, grain weight, and maize stover yield(Baral\u0026amp; Adhikari, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Moreover, the utilization of biofertilizers has demonstrated improvements in grain yield, stimulation of seed germination, bolstering seedling resistance against both biotic and abiotic stress, nitrogen fixation, and the production of phytohormones, thereby promoting better growth in maize plants(Mirjana Jarak, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2012\u003c/span\u003e).Rudolph et al., (\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) findings revealed that the application of plant growth promoting rhizobacteria on maize seeds notably elevated seed germination, biomass, and overall yield (LG, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Similarly,(Lhamo et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) observed that treating maize seeds with Azotobacter resulted in higher chlorophyll content within the leaves. This biofertilizer also notably improved the growth of Zea mays, thereby advocating its use, especially for enhancing maize production(Mirjana Jarak, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2012\u003c/span\u003e).Potdar et al. (\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) discovered that applying Azotobacter derived from lignite to sweet corn seeds resulted in heightened growth and increased yield of sweet corn. In a comparative study conducted by Mahato \u0026amp; Neupane (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) between Azotobacter and Trichoderma, along with other fertilizers, on maize growth, seeds treated with Azotobacter displayed significant improvements in parameters such as plant height, stem girth, dry shoot weight, root length and width, as well as root weight. Meanwhile, the application of Trichoderma exhibited either negative effects or negligible impact. Similar findings were also noted in two cultivars of Zea mays L., namely HUDAIBA (HD) and MUGTAMA45 (MG).Hence, the current research endeavors to isolate Azotobacter, create biofertilizer utilizing Azotobacter, and conduct trial experiments to assess its effectiveness as a biofertilizer for sweetcorn (\u003cem\u003eZea mays convar\u003c/em\u003e) cultivation.\u003c/p\u003e"},{"header":"Material and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Isolation of microorganism\u003c/h2\u003e \u003cp\u003eThe soil sample was collected from pee field within the CSA Campus. Soil samples were obtained from the rhizosphere area around Pea plants. These soil samples, enclosed in sterile plastic bags, were transported to the laboratory to isolate Azotobacter. A stock solution was prepared by combining 1 gram of soil sample with 9 milliliters of distilled water in a test tube. Subsequently, a series of dilutions were conducted: initially, a 10-fold dilution (10\u0026thinsp;\u0026minus;\u0026thinsp;1) was achieved by adding 1 milliliter of the stock solution to 9 milliliters of distilled water using a sterilized pipette. This process was repeated for subsequent dilutions, such as 10\u0026thinsp;\u0026minus;\u0026thinsp;2, 10\u0026thinsp;\u0026minus;\u0026thinsp;3, and so on, until reaching a 10\u0026thinsp;\u0026minus;\u0026thinsp;9 dilution factor. The goal of these serial dilutions in this experiment was to decrease the density of cell cultures to a more manageable concentration, ensuring that the colonies formed would be distinct and less dense.Following the serial dilution process, a specialized nitrogen-free Ashby mannitol media was prepared to isolate Azotobacter. Compostion of Ashby mannitol media are discussed in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e4\u003c/span\u003e. Subsequently, 100 microliters (0.1 ml) of diluted samples from 10\u0026thinsp;\u0026minus;\u0026thinsp;3, 10\u0026thinsp;\u0026minus;\u0026thinsp;5, 10\u0026thinsp;\u0026minus;\u0026thinsp;7, and 10\u0026thinsp;\u0026minus;\u0026thinsp;9 were transferred from the test tubes to the Ashby culture media using a sterilized pipette. These specific dilution factors were selected to decrease bacterial concentration, ensuring that the cultures grown in the plate would not be dense, facilitating the growth of distinct, pure colonies of the microbes. The culture was evenly spread on Ashby media using a spreader and then incubated at 30˚C for 7 days. After incubation, pure cultures were isolated by streaking the colonies grown on the culture plate onto a new culture plate containing Ashby media, followed by another incubation for 3 days at 30˚C. All procedures were conducted under Laminar Air Flow (LAF) to maintain aseptic conditions. The streak plate technique was utilized to separate colonies of pure Azotobacter culture from the mixed microbial population, effectively thinning out bacterial cells to ensure they were well-separated and adequately spaced.\u003c/p\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Characterization\u003c/h2\u003e \u003cp\u003eOnce bacterial strains were cultured in a pure environment, a small amount of the strain was utilized for gram staining. Following this, a motility test was carried out using the hanging drop method to assess the bacteria's motile properties. Additionally, biochemical tests including the methyl red test, VP test, starch hydrolysis, and catalase test were conducted to validate the identity of the bacterial strain.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Carrier Material\u003c/h2\u003e \u003cp\u003eThe components used as a carrier material for creating biofertilizer were derived from readily accessible local resources, including peat soil (P), forest organic matter obtained from leaf litter (F), and cow dung (C). These materials were gathered, dried under the sun, ground, sieved, and combined in a ratio of 1 part peat soil to 2 parts leaf litter to 2 parts cow dung, resulting in a final mixture of 4 kilograms. Subsequently, the carrier material underwent sterilization in an autoclave at 121\u0026deg;C for 15 minutes.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Inoculation\u003c/h2\u003e \u003cp\u003eTo prepare the microbial inoculants for mixing with the carrier material, a bacterial culture was cultivated in broth by transferring bacterial colonies into 500 milliliters of Azotobacter broth. This broth was then incubated in a shaking incubator at 30˚C, with an agitation speed of 200 revolutions per minute (rpm) for 6 days to foster mass culture. A total volume of 5000 milliliters of Azotobacter broth was generated. The biofertilizer formulation utilized the broth culture once the cell count of the inoculum reached 10\u003csup\u003e8\u003c/sup\u003e \u0026minus;\u0026thinsp;10\u003csup\u003e9\u003c/sup\u003e cells/ml. However, in this particular investigation, the growth rate of Azotobacter in the liquid medium (Azotobacter broth) was assessed by measuring the optical density using a spectrophotometer at a wavelength of 550 nm.When the optical density reached 1.334, the inoculum was combined with the carrier material. Different ratios were considered for mixing the carrier material with the broth culture: for every 1 kilogram of carrier material, sterile containers were used to mix 100 milliliters, 250 milliliters, and 400 milliliters of broth culture, respectively. Following thorough mixing, the carrier material containing the inoculant was allowed to cure for 6 days in a cool, dry place, kept away from direct sunlight and heat.\u003c/p\u003e \u003cp\u003eFollowing the curing period of six days, one gram from each combination was extracted to isolate the microbe, aiming to assess the bacterial density and confirm the presence of the microbial organism in the carrier material. The bacterial density was gauged using a spectrophotometer, revealing optical densities (OD) of 1.3, 1.4, and 1.5 at 550 nm for carrier materials treated with 100 ml, 250 ml, and 400 ml of the broth culture, respectively. The culture obtained underwent a biochemical test, confirming the presence of Azotobacter as the test yielded positive results for this specific microorganism.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.5 Soil Preparation\u003c/h2\u003e \u003cp\u003eAn initial trial was carried out using the prepared biofertilizer in a plastic container within a greenhouse setting. Soil samples were gathered from the garden area at HBTU Campus, Kanpur. These soil samples were subjected to air-drying, grinding, sieving (2 mm), and thorough mixing. The botany laboratory at Sherubtse College conducted qualitative assessments of soil parameters, including soil organic carbon, organic matter, porosity, water holding capacity, soil texture, soil conductivity, and pH. These analyses were conducted both before and after the application of the biofertilizer to assess its impact on soil properties.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e2.6 Experimental Design\u003c/h2\u003e \u003cp\u003eAll the procedures were conducted within a greenhouse environment using plastic pots situated near the botany laboratory at Chandrashekar Azad University, spanning from February to April 2023. The experimental setup encompassed 21 treatments arranged in a randomized block design, with three replicates for each treatment, and each pot represented a single replicate, totaling 63 pots. Each plastic pot, measuring 30x28 cm, was filled with approximately 5 kilograms of thoroughly mixed soil. Following soil filling, a recommended quantity of biofertilizer was added and mixed into the soil within the pots. Subsequently, seeds were sown. Additionally, a control experiment was established wherein seeds were sown in potted soil without the incorporation of any formulated biofertilizer, aimed at evaluating the efficacy of the biofertilizer. When required, the pots were watered with 250 milliliters of tap water. The details outlining the 21 different treatments are presented in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e1\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 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eDifferent types of treatment provided to the soil\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\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 \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSoil Treatments\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eReplica\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eAmount of Biofertilizer\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eUntreated Soil\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eST1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSeed Treatment\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eST2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eSeed treated\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eST3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100 ml, 100 g\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSoil was subjected\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eST4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100 ml, 200 g\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eto Biofertilizer\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eST5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100 ml, 300 g\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003etreatment before\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eST6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e250 ml, 100 g\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eplanting the seeds\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eST7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e250 ml, 200 g\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eST8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e250 ml, 300 g\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eST9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e400 ml, 100 g\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eST10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e400 ml, 200 g\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eST11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e400 ml, 300 g\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSoil treatment was\u003c/p\u003e \u003cp\u003edone two times:\u003c/p\u003e \u003cp\u003eFirst, before top\u003c/p\u003e \u003cp\u003eDressing after two\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eST12\u003c/p\u003e \u003cp\u003eST13\u003c/p\u003e \u003cp\u003eST14\u003c/p\u003e \u003cp\u003eST15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100ml, 100g\u003c/p\u003e \u003cp\u003e100ml, 200g\u003c/p\u003e \u003cp\u003e100ml, 300g\u003c/p\u003e \u003cp\u003e250ml, 100g\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWeeks of seed\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eST16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e250 ml, 200 g\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eplanting\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eST17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e250 ml, 300 g\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eST18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e400 ml, 100 g\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eST19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e400 ml, 200 g\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eST20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e400 ml, 300 g\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAfter addition of carrier matreial\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eST21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNo inoculants\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e2.7 Data collection and analysis\u003c/h2\u003e \u003cp\u003eAfter a month of sowing, data was collected on the height of the plants, the number of leaves on each plant, the length and width of the leaves, the stem girth, the number and length of the roots. The highest point of the uppermost leaf's arch, where the tip points downward, was used to measure the height of the plant. Leaf width was measured at the point where the leaf reached its maximum width, and leaf length was measured from the tip of the leaf to the point where it joins the leaf sheath. There was also a count of the leaves on each plant. Both the primary root's length and the total number of roots were measured. Using a ruler, the plant's height, width, length of leaves, and length of roots were measured in centimetres. A millimeter-based vernier calliper was used to measure the stem's girth.Graphpad Prism was utilised to analyse the replicated data. In a randomised complete block design, this was utilised to compute analysis of variance (ANOVA), standard errors, variance, and coefficient of variation. To determine if there were any differences amongst the 21 soil treatments, ANOVA was performed. To compare all potential pairwise treatments, Tukey's Multiple Range Test was used. A 5% level of significance (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) was used to estimate the significance of the differences between the treatment combinations.\u003c/p\u003e \u003c/div\u003e"},{"header":"Result and discussion","content":"\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e3.1 Isolation of microorganism\u003c/h2\u003e \u003cp\u003eUsing Ashby mannitol medium, a selective medium for Azotobacter, the nitrogen-fixing bacteria was successfully isolated, and their growth in the culture media confirms Azotobacter. Both morphological and biochemical tests were used to characterise the isolated bacteria. The colonies were mucoid, white, milky, round, uneven, and soft. The goal of identifying bacteria is to verify that the strain that was grown or obtained is the desired microbe. Isolating bacteria is necessary to obtain desired microbes for the study. The colonies were counted using a standard plate count method after they had formed on the culture media.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e3.2 Characterization\u003c/h2\u003e \u003cp\u003eGramme staining was used to verify that Azotobacter was present. These bacteria are gram-negative and have a safranin red colour. Azotobacter has an oval, rod-shaped pleomorphism. Since Azotobacter is a motile bacterium, the bacterial motility was tested using the hanging drop method. It was discovered that the material under the microscope was motile because rod- and oval-shaped bacteria could be seen moving in a particular kind of wet mount. Through biochemical characterization of Azotobacter, the isolated strain was verified; starch hydrolysis, methyl red, Voges-Proskauer (VP), and the catalase test all yielded positive results. The biochemical test result for Azotobacter is displayed in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003e3.3 Analysis of soil\u003c/h2\u003e \u003cp\u003ePrior to and following the treatment, the texture, organic carbon, organic matter, porosity, pH, and soil conductivity of the soil used in the pot experiment were examined. Sweetcorn is recognised to thrive in loam soil that has a pH ranging from 5.8 to 7 when organic matter is properly supplied. The data collected for soil properties both before and after treatments are displayed in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. The results are consistent with the research of Mahato\u0026amp;Neupane (2017), who found that applying biofertilizer enhances the physical characteristics of the soil and increases soil fertility and nutrient availability to plants. Additionally, organic matter increases the ability of microbes to hold onto water and gives them nutrients, which promotes microbial activity. According to Umesha et al. (2014), organic matter also increases the soil's ability to hold water and gives microbes nutrients, which promotes microbial activity and supports plant growth. Additionally, it was stated that biofertilizers contributed to the fertility of the soil. Usingbiofertilizer increases the amount of organic matter in the soil, which is a crucial sign of soil fertility.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003e3.4 Measurement of development plant\u003c/h2\u003e \u003cp\u003eFollowing a month of sowing, the data was recorded regarding various growth parameters. Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e2\u003c/span\u003e illustrates how the mean values are arranged in ascending order of the values.Contrary to some previous work done, this study showed that the plant height, stem girth, leaf length, and root length were the lowest. This could be because the seeds were treated differently with Azotobacter inoculants. In this study, seeds were kept in the shade for 30 minutes prior to sowing after being soaked in Azotobacter broth for 20 minutes without the use of any sticking agents.\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 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eImapct of biofertilizer on development of sweetcorn.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\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=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSoil Treatment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePlant height (cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStem girth(mm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNo. of leaves\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eLeaf length (cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eLeaf width (cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eRoot length(cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNo. of seed germinated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eDays taken to germinate\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eST1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e21.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e13.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e11.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e3.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e13.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eST2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e19.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e12.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e11.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e3.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e11.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eST3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e21.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e14.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e11.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e4.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e11.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eST4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e24.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e18.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e13.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e5.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e11.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5T5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e25.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e17.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e11.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e5.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e9.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eST6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e24.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e18.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e13.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e5.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e11.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eST7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e24.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e18.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e12.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e5.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e9.4\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eST8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e26.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e18.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e12.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e5.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e9.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eST9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e26.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e17.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e14.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e5.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e10.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eST10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e26.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e19.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e13.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e5.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e9.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eST11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e28.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e19.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e12.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e5.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e9.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eST12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e21.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e14.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e12.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e5.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e11.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eST13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e23.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e16.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e12.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e6.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e11.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eST14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e23.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e15.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e12.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e4.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e9.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eST15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e23.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e16.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e13.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e6.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e9.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eT16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e23.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e16.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e11.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e6.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e9.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eST17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e23.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e16.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e11.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e5.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e9.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eST18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e24.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e17.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e12.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e5.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e9.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eST19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e25.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e17.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e12.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e6.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e9.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eST20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e25.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e18.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e13.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e5.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e9.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eST21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e22.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e16.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e12.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e3.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e11.6\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\u003eThe results of this study indicate that the application of biofertilizers did not significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) affect the vegetative growth parameters. Out of all the treatments, soil treatment 11 (ST11) had the tallest plant (28.5 cm) and leaf length (19.5 cm). Soil treatment 2 (ST2) had the lowest plant height, indicating a detrimental effect over the control setup. ST2 yielded the highest number of leaves ever recorded. ST6 had the largest leaf width (2.2 cm), while ST21 had the smallest leaf width (1.7 cm). This led to an adverse effect on the control setup. According to similar research, leaf length was marginally greater in soil-inoculated Azotobacterbiofertilizerand lowest in seed-inoculated Azotobacter. In addition, Khushali et al. (2015) found that, in comparison to other treatments, leaves treated with Azotobacter seeds were smaller.\u003c/p\u003e \u003cp\u003eSoil treatment 5 (ST5) recorded the maximum stem girth (3.2 mm), soil treatment 6 (ST6) the leaf width (2.2 cm), soil treatment 9 (ST9) the root length (14.3 cm), and soil treatment 15 (ST15), 16 (ST16), and 19 (ST19) the maximum number of seeds that germinated. As shown in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e1\u003c/span\u003e, biofertilizer is applied to each treatment method in varying quantities. Some previous works by various authors has stated the increased rate of organic fertiliser is what caused the increase in plant parameters, which may be explained by the fact that it contains a good amount of nutrients both (micro and macro nutrients) that is essential for the growth and development of plants.In addition, Mirza et al. (2000) reported that biofertilizers facilitate the synthesis of substances that promote plant growth, such as auxin, which raises lethal fibres and enhances plant growth in addition to fixing nitrogen. The lack of significant variation in plant parameters between those treatments may be due to the plant parameters' high genetic influence, which has no discernible impact on the application of biofertilizers.\u003c/p\u003e \u003cp\u003eThe application of biofertilizer had a significant impact on parameters such as the number of days taken for germination. The number of seeds that germinated varied amongst treatments, but there was no statistically significant difference; however, ST15, ST16, and ST19 showed the highest levels of seed germination. The maximum number of days required for germination in ST1 was 13, and the lowest number of days required was 8 in soil treated with biofertilizer (ST8, ST10, ST11, ST14, and ST17\u0026ndash;T20). However, the number of days required for germination following the application of biofertilizer was statistically significant with the control setup (ST1). The outcome was consistent with the previous researches, that shown using biofertilizer increased seed germination.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003e3.5 A comparative analysis of soil treated once (ST3\u0026ndash;ST11) twice and (ST12\u0026ndash;ST20):\u003c/h2\u003e \u003cp\u003eThe means in the same column that are followed by the same letters do not differ significantly from one another. The means from ST3\u0026ndash; ST11 are represented by ST-1, and the means from T12\u0026ndash;T20 are represented by ST-2.Nine of the twenty-one treatments (ST3\u0026ndash;ST11) were single treatments, wshile the other nine treatments (ST12\u0026ndash;ST20) are double treated, meaning that the soil is treated with biofertilizers twice: once during seed sowing and again two weeks later when the plants receive a topdressing of biofertilizers. Since maize is a heavy feeder crop that needs a lot more nutrients than other crops, it received two treatments. Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e3\u003c/span\u003e presents the mean values for soil treated with single treatment (ST3-ST11) and double treatment (ST12- ST20). There was a statistically significant difference in plant height, stem girth, and leaf length between the treatments. Other plant characteristics, however, including the quantity of leaves and roots, the width and length of the leaves, the amount of chlorophyll, the number of seeds that germinated, and the time it took for those seeds to germinate, did not differ significantly between the two treatments.\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 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ecomparative analysis of soil treated once (ST3- ST11) and twice (ST12-ST20)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\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=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSoil Treatment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePlant height (cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStem girth(mm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNo. of leaves\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eLeaf length (cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eLeaf width (cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eRoot length(cm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNo. of seed germinated\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eDays taken to germinate\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eST-1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e25.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.789\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e18.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e12.77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e5.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e9.88\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eST-2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e23.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e273\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3,677\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e16.64\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e12.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e5.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e9.57\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\u003eHowever, compared to double treatment, single treated soil produced higher plant growth values. This could be because the plants were uprooted for analysis after two weeks of receiving biofertilizer topdressing. It's possible that the microbial activity and nutrients needed to support plant growth weren't completed in the allotted two weeks. Additionally, the amount, timing, and technique of topdressing may have varied from other methods because Mahato \u0026amp; Neupane (2017) used the dibbling method to topdress fertiliser 26 days after sowing. Others suggested that top dressing be applied when the maize plant reaches a height of approximately 18 inches, or six to eight weeks following the plant's emergence which gives effect of topdressing. Nevertheless, topdressing was carried out in the current study following two weeks of seeding.\u003c/p\u003e \u003c/div\u003e\u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eList of Chemical Used In Ashby Mannitol Media\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"2\"\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 \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIngredients (gm per 100 ml)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003ecomposition\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMannitol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2gm\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDi potassium phosphate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.02gm\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMagnesium sulphate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.02gm\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSodium chloride\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.02gm\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePotassium sulphate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.01gm\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCalcium carbonate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.5gm\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAgar- agar\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.5gm\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis study contributes to the growing body of knowledge on importance of biofertilizers as an environmentally friendly substitute for conventional fertilisers. It was clear from a thorough examination of numerous plant growth characteristics that the sweetcorn plants benefited from the application of biofertilizer. According to the results, biofertilizer may be able to raise sweetcorn crops' overall productivity. Additionally, the statistical analyses offered a strong basis for determining the importance of variations among different treatments. These analyses strengthened the validity of the observed results by giving the study a quantitative component. As we move forward, it is crucial to acknowledge the potential implications of incorporating biofertilizer into sweetcorn cultivation practices. Nonetheless, the samples treated with the specially blended biofertilizer had fewer days for the seeds to germinate. So, more research focusing on measuring the yield components is required to comprehend the efficacy of various treatments on \u003cem\u003eZea mays convar\u003c/em\u003e. The favourable results reported in this study provide opportunities for additional investigation and use of biofertilizer in various agricultural contexts.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003e8.1 Ethical Approval\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e8.2 Consent to Participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll participants involved in this study provided informed consent before participating, understanding the purpose and procedures involved.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e8.3 Consent to Publish\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eI confirm that all co-authors have reviewed and given their consent for the publication of this research paper.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e8.4 Authors Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe would like to express sincere gratitude to Shravan kumar for their invaluable guidance and support in shaping manuscript to be published. Prateek Mishrahas contributed significantly to the conception, design and writing of the research. Anjali Singh has contributed in data collection and lab work and Surbhi Sharma has contributed in the data analysis, and interpretation of the research.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e8.5 Funding\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e8.6 Competing Interests:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing interests that could influence the objectivity, integrity, or validity of the research.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAgarwal P, Gupta R, Gill IK (2018) Importance of biofertilizers in agriculture biotechnology. 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Nat Biofertil Sustanainability Agricultural. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003eAugust. https://doi.org/10.1007/978-3-030-18933-4\u003c/span\u003e\u003cspan address=\"August. 10.1007/978-3-030-18933-4\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSuhag M (2016) \u003cem\u003ePotential of Biofertilizers to Replace Chemical Fertilizers\u003c/em\u003e. \u003cem\u003eApril\u003c/em\u003e, 1\u0026ndash;6. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.17148/IARJSET.2016.3534\u003c/span\u003e\u003cspan address=\"10.17148/IARJSET.2016.3534\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSumbul A, Ansari RA, Rizvi R, Mahmood I (2020) Saudi Journal of Biological Sciences Azotobacter: A potential bio-fertilizer for soil and plant health management. Saudi J Biol Sci 27(12):3634\u0026ndash;3640. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.sjbs.2020.08.004\u003c/span\u003e\u003cspan address=\"10.1016/j.sjbs.2020.08.004\" 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":"Bio-fertilizer, Soil fertility, Azotobacter, Zea mays convar","lastPublishedDoi":"10.21203/rs.3.rs-3854929/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3854929/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eExcessive utilization of agrochemical fertilizers significantly leads to the decline of soil quality and its flora, it also affect human health. A material called bio-fertilizer contains living microorganisms that are advantageous to the growth and development of plants.Due to the impact of biofertilizers on enhancing crop yield, there is a growing focus on exploring beneficial microbes as biofertilizers within the agricultural domain. Consequently, this study was carried out to assess the efficacy of Azotobacterbiofertilizer on Sweet Corn (\u003cem\u003eZea mays convar\u003c/em\u003e).An experiment in a controlled artificial greenhouse setting involved 21 different treatments utilizing varying quantities of inoculants and carrier materials to create biofertilizer. The carrier material, derived from locally accessible raw materials such as cow dung, leaf litter, peat soil, lignite, charcoal, and vermiculite, serves as a substrate capable of hosting microbial inoculants and maintaining their viability for a specific duration, while also being easily accessible to farmers. Following the sowing of sweetcorn seeds for a month, the resulting germinated plants underwent analysis for parameters including plant height, stem girth, leaf count, length and width of leaves, as well as root length.The quantity of seed germination and the duration required for seeds to germinate were also recorded. Out of the 21 treatments, ST11, comprising 400 ml of inoculants combined with 300 g of carrier material, displayed higher values for plant parameters compared to the other treatments, although the statistical significance among all treatments was minimal. Each plant parameter responded differently to the various treatments. Contrary to the findings of others, the seeds immersed in microbial inoculant (ST2) did not notably enhance plant growth except for the number of leaves per plant. To validate the effectiveness of the formulated biofertilizer, a control was established: one set of seeds were sown solely in pot soil, while another contained only carrier material (ST21) without any microbial inoculant, which did not facilitate plant growth. Overall, plants treated with biofertilizer exhibited a positive response in terms of plant growth compared to the control group.\u003c/p\u003e","manuscriptTitle":"Efficacy Testing of Bio-fertilizer Based on Azotobacter on Sweet Corn (Zea mays convar)","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-03-06 09:11:20","doi":"10.21203/rs.3.rs-3854929/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":"d8ff7c8b-072a-4a06-938e-112c6dda8241","owner":[],"postedDate":"March 6th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-03-18T20:02:59+00:00","versionOfRecord":[],"versionCreatedAt":"2024-03-06 09:11:20","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-3854929","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3854929","identity":"rs-3854929","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}