Bio-composition for enhancing nutrient use efficiency in plants

Abstract

The present invention discloses composition and formulations to enhance nutrient use efficiency in plants. The composition is designed to enhance nutrient uptake, optimize fertilizer utilization. It comprises brown seaweed extract combined with selected botanical extracts, processed using a water-ethanol cold percolation extraction method that preserves key bioactive compounds such as phenolics, flavonoids, tannins etc. This formulation improves nutrient use efficiency by increasing nutrient uptake. Field trials across different plant species confirm a 20% reduction in fertilizer usage without yield loss, demonstrating its efficacy in improving nutrient uptake and plant metabolism. By reducing fertilizer dependency, the invention promotes sustainable agriculture while minimizing environmental impact. Its versatile application as a foliar spray, root drench, or fertilizer additive makes it adaptable to diverse crop types and farming systems, providing a scalable, eco-friendly alternative to conventional fertilizer-based growth enhancement methods.

Description

DescriptionTitle of Invention: BIO-COMPOSITION FOR ENHANCING NUTRIENT USE EFFICIENCY IN PLANTSTechnical Field

[0001] The invention relates to the field of agricultural biotechnology, plant nutrition, and sustainable agriculture. In particular, the invention relates to compositions for enhancing nutrient use efficiency in plants comprising brown seaweed (Durvillaea potatorum) and botanical extracts obtained by a water-ethanol cold percolation process, designed to improve nutrient use efficiency in plants, thereby reducing dependence on synthetic fertilizers while maintaining crop yields.Background Art

[0002] Considering the significant challenges in modern agriculture, including declining nutrient use efficiency, excessive fertilizer dependency, and environmental concerns such as nitrogen leaching and greenhouse gas emissions, there is an urgent need for sustainable and effective plant nutrition solutions. Globally nutrient use efficiency is about 30% and in some regions like India, Africa it is even lower (20%). Additionally, factors such as poor soil health, suboptimal root development, and abiotic stresses like drought and salinity further impact crop productivity, thereby necessitating innovative approaches to enhance nutrient uptake and plant resilience.

[0003] Seaweed-based plant compositions have gained attention for their potential in improving plant growth, stress tolerance, and nutrient absorption. However, most commercially available formulations rely on Ascophyllum nodosum and conventional extraction techniques such as fermentation or enzymatic hydrolysis. These methods, while beneficial, may degrade sensitive bioactive compounds, limiting the full potential of the extract.

[0004] Global agriculture faces an acute challenge in nutrient management. Fertilizer inputs, particularly nitrogen, phosphorus, and potassium (NPK), remain highly inefficient. Global average NUE is -30%, while in regions such as India and Sub-Saharan Africa, NUE is as low as 20%. Excessive fertilizer use contributes to soil degradation, nitrogen leaching, eutrophication, and greenhouse gas emissions. Declining soil organic carbon and poor microbial diversity exacerbate inefficiencies. Over years Nitrogen use efficiency has stagnated, phosphorous use efficiency is low and potassium use efficiency has in fact decreased. Chemical nitrification inhibitors and stabilizers offer limited solutions and often disrupt soil microbiota. Conventional fermented seaweed formulations (Ascophyllum nodosum) provide some biostimulant benefits but degrade thermolabile bioactive during extraction.

[0005] The agricultural industry is increasingly prioritizing sustainable alternatives that optimize nutrient use efficiency (NUE) while reducing the reliance on chemical fertilizers. The excessive use of synthetic fertilizers not only leads to economic inefficiencies but also contributes to long-term soil degradation, high carbon footprint and environmental pollution. Therefore, there is a growing demand for formulations that enhance nutrient absorption, nutrient uptake, improve plant metabolism, and promote sustainable agricultural practices.

[0006] The majority of existing patents focus on Ascophyllum nodosum as the primary seaweed species for biostimulant or nutrient uptake enhancement. These products are widely documented for improving plant health and mitigating abiotic stress, with some secondary claims about improving NUE. Extraction methods commonly used are fermentation, enzymatic hydrolysis, or heat-based solvent extraction. Examples: US Patent No. US8439896B2: "Seaweed-based biostimulants for nutrient efficiency improvement in crops.", W02020054032A1: "Seaweed extract-based fertilizer additive for reducing nitrogen leaching and improving nutrient uptake."

[0007] Some patents combine seaweed extracts with botanical or microbial components to improve NUE, but these are relatively rare and do not specifically emphasize the use of cold percolation or non-Ascophyllum seaweed species. Example: W02021103920A1: "Biostimulant compositions containing plant and seaweed extracts for enhancing fertilizer use efficiency." US8439896B2 describes a seaweed-based plant bio stimulant aimed at improving nutrient efficiency in crops, primarily utilizing Ascophyllum nodosum extracts. However, it does not explore the use of Durvillaea potatorum, and potential for enhanced nutrient uptake.

[0008] There is therefore a critical need for a sustainable bio-composition that enhances nutrient uptake, assimilation, and use inside plants, supports abiotic stress resilience (drought, salinity, heat), reduces fertilizer dependency while maintaining or increasing yield and preserves delicate secondary metabolites (phenolics, flavonoids, tannins) critical to nutrient use efficiency.Summary of Invention

[0009] Before the present invention is described, it is to be understood that the present invention is not limited to specific methodologies and materials described, as these may vary as per the person skilled in the art. It is also to be understood that the terminology used in the description is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention.

[0010] The present invention relates to bio-compositions to enhance nutrient use efficiency in plants. The composition mainly comprises an extract of brown seaweed (Durvillaeapotatorum) and one or more botanical extracts for enhancing nutrient uptake and reducing fertilizer dependency in crops.

[0011] The invention further includes a unique water-ethanol cold percolation extraction process used for obtaining bioactive compounds from the seaweed and botanical extracts. This method ensures the preservation of delicate bio actives, such as flavonoids, Phenolics (Gallic acid and Syringic acid) and other secondary metabolites, which contribute to enhanced nutrient use efficiency and nutrient uptake.

[0012] By combining a novel seaweed species with carefully selected botanical extracts and utilizing an innovative extraction method, the present invention offers a breakthrough in improving nutrient use efficiency by targeting not just nutrient availability but also uptake and use efficiency, reducing chemical fertilizer dependency, and promoting sustainable agricultural practices with more nutrient dense produce.Advantageous Effects of Invention

[0013] The advantages of the present invention are as follows:• The formulation improves nutrient absorption and utilization, leading to better crop performance with reduced fertilizer dependency.• By reducing fertilizer dependency by up to 20%, the invention supports cost-effective and environmentally responsible farming practices.• Improves photosynthetic efficiency and optimizes root architecture, leading to stronger and more resilient plants.• The formulation is safe, stable, non-toxic, and eco-friendly, making it suitable for diverse Agro-ecosystems.• It is fully compatible with standard agronomic practices and seamlessly integrates into existing farming operations.• The unique blend of brown seaweed (Durvillaea potatorum) and select botanical extracts enhances root architecture, plant metabolism, and overall growth efficiency.• The water-ethanol cold percolation extraction method preserves key bioactive compounds, including flavonoids, phenolics, tannins and other secondary metabolites, ensuring maximum efficacy in plant stress tolerance and nutrient uptake.• In contrast to conventional fermentation or heat-based extraction processes that may degrade essential bio actives, the cold percolation method retains their potency, leading to improved plant resilience and long-term effectiveness.• The formulation enhances nutrient uptake, use, promoting photosynthetic efficiency, particularly under low nutrient conditions.• Multiple field trials across various crops, including rice, tomato, capsicum, etc. have shown that fertilizer usage can be reduced by 20% without any yield loss, making the formulation a cost-effective and sustainable solution.• By improving nutrient use efficiency, the formulation reduces nitrogen leaching, soil degradation, and environmental contamination, supporting eco- friendly and sustainable agricultural practices.• The formulation can be applied as a foliar spray, root drench, or as a fertilizer additive, making it adaptable to different crop types and farming conditions.• By providing a stable and scalable alternative to synthetic fertilizers, the invention contributes to improved soil health, reduced dependency on chemical inputs, and enhanced productivity in modern agriculture.• Collectively, these advantages align with the principles of sustainable agriculture and climate-smart farming, offering a practical solution for enhancing crop performance while minimizing environmental impact.Brief Description of Drawings

[0014] A complete understanding of the present invention may be made by reference to the following detailed description, which is to be taken in conjunction with the accompanying drawings. The accompanying drawings, which are incorporated into and constitute a part of the specification, illustrate one or more embodiments of the present invention and, together with the detailed description, serve to explain the principles and implementations of the invention.Fig.l

[0015] [Fig.l] depicts the graphs illustrating the tank- mix compatibility of the composition with a wide range of foliar fertilizers and agrochemicals.

[0016] [Fig.2]

[0017] [Fig.2] depicts the graph that show the consistency in the composition batches produced over 3 years.

[0018] [Fig.3]

[0019] [Fig.3] depicts the graph that exhibits high formulation stability.

[0020] [Fig.4]

[0021] [Fig.4] depicts the comparative graphs of different parameters for nutrient uptake efficiency improvement; and

[0022] [Fig.5]

[0023] [Fig.5] is a representation that highlights the mechanism of working of the bio composition.Description of Embodiments

[0024] Before the present invention is described, it is to be understood that this invention is not limited to methodologies described, as these may vary as per the person skilled in the art. It is also to be understood that the terminology used in the description is for the purpose of describing the particular embodiments only and is not intended to limit the scope of the present invention. Throughout this specification, the word “comprise,” or variations such as “comprises” or “comprising,” will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps. The use of the expression “at least” or “at least one” suggests the use of one or more elements or ingredients or quantities, as the use may be in the embodiment of the invention to achieve one or more of the desired objects or results. Various embodiments of the present invention are described below. It is, however, noted that the present invention is not limited to these embodiments, but rather the intention is that modifications that are apparent are also included.

[0025] The present invention discloses bio-compositions to enhance nutrient use efficiency in plants. The compositions comprise of extracts of brown seaweed (Durvillaea potatorum) and one or more botanical extracts, in varying concentrations and proportions, prepared using a water-ethanol cold percolation extraction process.The compositions enhance nutrient use efficiency in plants by improving nutrient uptake, optimizing metabolic activity thereby allowing for a 20% reduction in fertilizer usage without compromising crop yield. The compositions of the present invention synergistically combine bioactive plant metabolites to optimize nutrient assimilation. In contrast to the conventional chemical nitrification inhibitors that may disrupt soil microbiota, this formulation ensures sustainable nutrient management without affecting soil microflora and promoting plant- microbe interactions. The present invention provides bio-compositions that are designed to improve nutrient use efficiency by facilitating the conversion, uptake, and assimilation of nutrients in plants and resulting into a nutrient dense produce.

[0026] The raw materials used in the composition comprises of Brown seaweed that is Durvillaea species that is selected from Durvillaea antarctica, Durvillaea potatorum Durvillaea poha, Durvillaea willana, Durvillaea fenestrate, Durvillaea amatheiae Durvillaea incurvate Durvillaea chathamensis etc. Selected botanicals containing flavonoids, polyphenols, flavonoids, phenolic acids (including gallic acid and syringic acid), tannins are selected from Salvia indica, Salvia officinalis, Triticum aestivum, Sorghum propinquum, Hordeum vulgare, Trigonella foenum-graecum, Trigonella corniculata, Trigonella caerulea, Medicago sativa, Medicago truncatula, Medicago polymorpha, , Emblica officinalis (Phyllanthus emblica), Phyllanthus amarus,Phyllanthus niruri, Phaseolus vulgaris, Phaseolus lunatus, Phaseolus acutifolius, Ocimum tenuiflorum (O. sanctum), Ocimum gratissimum.

[0027] The source and geographical origin of the biological materials used in the invention are as follows:The source and geographical origin of other botanical extracts is the same as Indian sage.

[0028] According to the embodiment of the present invention, the drying and pulverization of the seaweed and botanical biomass comprises of the following steps. Freshly harvested seaweed biomass is first rinsed with clean water to remove surface salts, sand, and epiphytes. Freshly harvested botanical biomass is first rinsed with clean water to remove sand, and other impurities. The cleaned biomass of both is then subjected to solar shade drying under ambient conditions. The drying process is conducted in a well- ventilated covered structure that allowed diffused sunlight and airflow while preventing direct UV exposure, ensuring preservation of bioactive compounds. The temperature during drying is maintained below 45 °C, with continuous monitoring to avoid degradation of thermolabile secondary metabolites. Drying is continued until the moisture content of the biomass is reduced to <10%, or until the biomass became crisp and brittle to touch. Once fully dried, the seaweed and botanical biomass is milled using a hammer mill and passed through a 40-mesh sieve to obtain a fine, uniform powder suitable for solvent extraction. The resulting powder is stored in airtight, light-resistant containers at room temperature until use.

[0029] According to the embodiment of the present invention, the extraction is conducted using cold percolation method with water-ethanol solvent (60:40 v / v) to maximize recovery of active metabolites. The extraction process employs a mixed water-ethanol solvent system at a volumetric ratio of approximately 60:40 (v / v), selected to enablebalanced extraction of both hydrophilic and lipophilic bioactive constituents from the biomass. The solvent is applied at a ratio of about 10:1 relative to the dried plant material to ensure optimal metabolite recovery. Extraction is conducted at an ambient temperature of approximately 25 °C in order to minimize thermal degradation of heat- sensitive metabolites. The extraction period may range from about 12 hours to 14 days, permitting exhaustive recovery of soluble constituents depending on the material and process scale. Following extraction, solvent removal is carried out by rotary evaporation under reduced pressure at temperatures between 40 °C and 45 °C so as to preserve the bioactivity of the recovered extract. The resulting concentrate is subsequently standardized to contain not less than 1000 ppm polyphenols and not less than 1000 ppm tannins, providing assay-based consistency in the final composition.

[0030] The extraction approach described herein provides several advantages over fermentation-based processes. Notably, it enables significantly higher recovery of phenolic and flavonoid constituents, delivering a more concentrated and bioactive extract. The method also results in minimal co-extraction of alginates, mannitol, and other polysaccharide impurities that are typically elevated in fermentation outputs. In addition, the controlled solvent-based process affords superior batch-to-batch reproducibility, ensuring consistent chemical profiles and standardized bioactive levels across production runs.

[0031] According to the embodiment of the present invention, the bio composition comprises the extract of Durvillea at approximately 70% w / v that serves primarily as a source of phenolic and tannin-rich constituents, which contribute antioxidant activity and facilitate modulation of indole- 3 -acetic acid (lAA)-related signaling pathways, melatonin signaling, Spermine signaling and Arginine metabolism for uptake and use efficiency and the selected botanical extract (such as Salvia indica, Salvia officinalis, Triticum aestivum, or others) at approximately 20% w / v that functions chiefly as a nitrogen-buffering component, supporting improved nitrogen-use efficiency (NUE), enhanced nitrogen storage within plant tissues, and subsequent nitrogen remobilization during key developmental stages. Together, these extracts provide synergistic benefits that support plant growth, stress tolerance, and nutrient optimization.

[0032] According to the embodiment of the present invention, the bio composition comprises a defined spectrum of bioactive compound classes that contribute to improved nitrogen-use efficiency in plants. The phenolic fraction, which may range from approximately 10 to 50,000 ppm (w / w), includes compounds such as gallic acid and syringic acid and provides antioxidant capacity as well as nitrogen-buffering effects. Flavonoids, present at about 10 to 20,000 ppm, including quercetin and luteolin, support auxin transport modulation and enhance early root development. Tannins may be included at concentrations between roughly 500 and 20,000 ppmto facilitate reactive oxygen species (ROS) scavenging and improve plant stress tolerance. Polyamines, such as putrescine derivatives, may occur at levels of 50 to 5,000 ppm and function in enzyme stabilization and NUE enhancement. Additionally, amino acid derivatives — including arginine and branched-chain amino acids — may be present at concentrations of about 100 to 5,000 ppm, where they act to support nitrogen storage and subsequent remobilization during critical growth stages.

[0033] According to the embodiment of the present invention, the formulated bio composition exhibits high stability and consistency under standard storage and application conditions. In particular, the composition demonstrates a shelf life of at least four years when maintained within a temperature range of approximately 5-45 °C, without evidence of degradation or loss of functional activity. Batch-to- batch reproducibility is ensured through spectral fingerprinting techniques, which confirm the uniformity of the chemical profile across production lots. Heavy-metal contaminants, including but not limited to lead, cadmium, arsenic, and mercury, are maintained below detectable limits (typically <0.1 ppm), ensuring regulatory compliance and product safety. Toxicological assessments further indicate that the composition is non-toxic to humans, animals, pollinators such as bees, earthworms, and a wide range of aquatic species, rendering it suitable for safe agricultural and environmental use.

[0034] Compatibility: According to the embodiment of the present invention, the bio composition is designed for broad-spectrum compatibility with commonly used fertilizers, micronutrient blends, and crop protection products. Laboratory and fieldscale evaluations have confirmed no adverse interactions, ensuring consistent efficacy and performance when applied in standard agronomic programs.

[0035] The bio composition of the present invention is readily miscible in water and demonstrates excellent tank-mix compatibility with a wide range of foliar fertilizers and fertilizers, including agrochemicals. Concentrated, ready-to-use doses formulation were mixed with fertilizers, micronutrients, insecticides, and pesticides to assess tankmix compatibility. The mixtures were evaluated for any changes in their spectral signatures at 0, 2, 4, and 6 hours after mixing as illustrated in [Fig.l][Table 1]

[0036] Stability and consistency: The scale-up and manufacturing process enables production of a highly reproducible bio formulation with a consistent and well-defined composition. For this spectral signature of separate batches from three separate years was done. The graph as illustrated in [Fig.2] reveals very high adherence to the spectral signature with very minute variation indicating consistency in batches produced over 3 years, this represents a significant advancement compared to fermented Ascophyllum- based biostimulants, which often exhibit considerable batch-to-batch variability.Achieving consistent formulation quality is critical, as it forms the foundation for delivering reliable and reproducible field performance.

[0037] Shelf life: The formulation exhibits high formulation stability with no significant phase separation, precipitation, or viscosity changes under storage temperatures between 5 °C and 45 °C. For this spectral signature of a single batches was done over a period four separate years was done. The graph as illustrated in [Fig.3], reveals very high adherence to the spectral signature with almost no variation. Its active components retain biological efficacy for a minimum of 4 years under recommended storage conditions.

[0038] Heavy Metal reduction: The extraction method of the present invention also significantly reduces the heavy metal content in the final bio combination formulation compared to traditional fermentation-based Ascophyllum biostimulants, enhancing both product safety and environmental sustainability as shown in Table 2.[Table 2] << < < <<<

[0039] Toxicity: Toxicological assessments confirm that the present bio composition biostimulant is non-toxic to humans, animals, and beneficial non-target organisms when used according to label directions. The formulation is classified as low-risk, with no reported phytotoxicity on tested crops as illustrated in TABLE 3. It meets safety standards for biostimulant use under prevailing agricultural regulations.[Table 3]

[0040] Application: Materials and Methods: The primary mode of delivery of the composition of the present invention can include foliar spray, fertigation, and as fertilizer additive, ensuring efficient uptake and maximum efficacy across diverse crops. For field application, comparative trials conducted across different crop species, including rice, tomato, capsicum etc. have shown substantial improvements in nutrient uptake efficiency, reduced nutrient leaching, and enhanced overall crop yield. The formulation promotes activation of plant mechanisms like ABC transporters which play a crucial role in nutrient uptake by actively transporting a wide variety of essential nutrients, Hormone response pathways, Plant-microbe interaction. Better availability and use of nutrients boost photosynthesis in plants resulting in better yield even under reduced fertilizer application.

[0041] The bio composition may be applied using multiple agronomically compatible delivery modes. In certain embodiments, it is administered as a foliar spray at aconcentration of approximately 3-5 mL per liter of water, enabling rapid uptake through leaf tissues. In other embodiments, the composition is delivered via fertigation systems at application rates of about 800-1000 mL per acre, providing uniform distribution through irrigation channels and facilitating root- zone assimilation. The formulation may also be incorporated as an additive to conventional granular or liquid NPK fertilizers at inclusion levels ranging from approximately 2% to 5% w / w, thereby enhancing nutrient efficiency and complementing standard fertilization programs.

[0042] 1. Rice -Rice, Variety DRR Dhan 65 was grown in the experimental plots of Indian Institute of Rice Research, Hyderabad, Location: Rajendranagar Farm, ICAR- IIRR, Hyderabad during Kharif’23 & Rabi’23-24. Each Plot Size was of 6.6 X 3.2 m and plants were sown at a spacing of 20 X 15 cm. According to ICAR the recommended fertilizer Dose (RDF) is of N:P:K:ZnSO4- 100:60:40:20 kg / ha. For this experiment fertilizer Dose applied was 50,75 &100% of RDF solely and with the present seaweed formulation to assess the improvement in nutrient uptake efficiency. The formulation was applied as foliar spray at 20 & 50 DAT at 500 & 1000 ml per acre doses. In total there were 12 treatments with 3 replicates. Each replicate had 704 plants and 25344 plants for entire experiment.

[0043] Parameters assessed: Plant height, SPAD, ND VI, tiller number, panicle number, grains per panicle, grain yield, straw yield, NPK content in leaf, stem and soil, available Fe & Zn in leaf and soil and NUE indices

[0044] Statistical Analysis: At least 704 plants replicates per treatment group. Data analyzed using ANOVA, followed by Tukey’s HSD post-hoc test (p < 0.05

[0045] The nutrient uptake efficiency improvement is illustrated in FIG. TABLE 4 and 5 below:[Table 4][Table 5]

[0046] In field evaluations, the nutrient-efficiency formulation produced substantial improvements in crop performance and soil nutrient dynamics as illustrated in [Fig.4] Grain yield increased by approximately 22.3% even when fertilizer inputs were reduced by 20% relative to the recommended dose of fertilizers (RDF 100%), demonstrating enhanced productivity under lower nutrient supply. Plants treated with the formulation exhibited elevated concentrations of nitrogen, phosphorus, and potassium, together with notable micronutrient enrichment, including approximately 16% higher iron and 22% higher zinc accumulation in harvested grain. Soil analyses at harvest further indicated improved nutrient retention, with available nitrogen increased by roughly 50%, available phosphorus by about 60%, and available potassium by approximately 25% compared to untreated controls. Correspondingly, nutrient- use-efficiency metrics showed marked enhancement: agronomic efficiency (AE) increased from 10.1 to 18.5 kg of grain per kilogram of nutrient applied, representing an improvement of approximately 83%, while recovery efficiency (RE) increasedfrom 49.7% to 107.4%, indicating substantially greater nutrient uptake and markedly reduced nutrient losses.

[0047] ROI Increase[Table 6]

[0048] Tomato: Tomato, Variety Big Beef was used to study impact of formulation in improving NUE in Jul-Nov’23 at San Luis Ranch farm California, USA. Total area of study 14X 7 Ft, with a spacing: 7 X 1.5 Ft, 20 plants per replicate, 1100 plants for entire experiment. For NUE study 100 & 80% Grower standard practice of Fertilizer dose was used. Application method was Foliar Spray at 33 & 68 Days after Planting at a Low rate of 400 ml / acre (2 ml / L), Mid-rate - 800 ml / acre (4 ml / L), & high rate- 1600 ml / acre (8 ml / L)[Table 7]

[0049] In the agronomic evaluations, the nutrient-efficiency formulation delivered significant yield and economic benefits even under reduced fertilizer inputs. When fertilizer application was decreased by 20% relative to the Grower Standard Practice (GSP 100%), crop yield increased by approximately 6.6%, while yields were elevated by about 33.39% compared to the GSP 80% regimen. These improvements translated directly into higher farm returns: growers realized an estimated additional income of $1,119 per acre compared to GSP 100%, and an increase of approximately $4,873 per acre relative to GSP 80%. The formulation also enhanced crop quality parameters. The number of marketable fruits increased by approximately 15.7% compared to GSP 80%, and total marketable fruit weight rose correspondingly — by about 6.6% relative to GSP 100% and by approximately 33.4% relative to GSP 80%. Collectively, these results demonstrate that the formulation enables substantial productivity and profitability improvements even under reduced fertilizer input conditions.

[0050] Bell peppers- Bell Pepper, Variety California was used to study impact of formulation in improving NUE in Jul-Nov’23 at San Luis Ranch farm California, USA. Total area of study 14 X 7 Ft, with a spacing: 6.6 X 1 Ft, 28 plants per replicate, 1512 plants for entire experiment. For NUE study 100 & 80% Grower standard practice of Fertilizer dose was used. Application method was Foliar Spray at 40 & 70 Days after Planting at a Low rate of 400 ml / acre (2 ml / L), Mid-rate - 800 ml / acre (4 ml / L), & high rate- 1600 ml / acre (8 ml / L).[Table 8]

[0051] With a 20% reduction in fertilizer relative to GSP 100%, crop yield increased by approximately 2.8%, and yields were enhanced by about 25.8% when compared to GSP 80%. These improvements translated into measurable financial gains, providing an estimated additional income of $508.7 per acre under the 20% fertilizer-reduction regime and approximately $3,880.6 per acre relative to GSP 80%. Quality parameters also improved substantially: the number of marketable fruits increased by roughly 12.6% with 20% less fertilizer and by approximately 42.4% compared to GSP 80%. Similarly, total marketable output rose by about 2.8% relative to GSP 100% and by nearly 25.8% compared to GSP 80%. Collectively, these results confirm that the formulation delivers both productivity and profitability gains while enabling meaningful reductions in fertilizer use.

[0052] Fertilizer Coating: The composition is suitable for application as a spray-coating onto finished fertilizers or water-soluble fertilizer blends. The process is streamlined, scalable, and requires minimal modifications to existing manufacturing or blending operations. The finished water-soluble fertilizer blend is placed into a standard industrial blender, mixer, or coating drum to ensure uniform tumbling prior to application of the bioactive formulation. An amount of 25-50 ml of the composition per acre equivalent is sprayed onto the fertilizer while the material is continuously tumbled. This ensures uniform and consistent coating of the fertilizer granules or pills with the bioactive formulation.

[0053] After coating the monoammonium phosphate (MAP) fertilizer with the nutrient use efficiency (NUE) formulation of the present invention at the recommended coating rate, the finished product was evaluated for its physical and chemical integrity. The coated MAP was assayed against the corresponding Fertilizer Control Order (FCO) specifications for MAP.

[0054] Analytical assessment confirmed that all physical and chemical parameters of the coated material remained within the permissible limits prescribed under FCO, with no deviation from the baseline MAP values. The coating process did not alter nutrientconcentration, granule structure, flowability, moisture content, or any regulated specification. The coated MAP maintained full compliance with FCO, demonstrating compatibility of the NUE formulation with standard fertilizer materials. As illustrated in TABLE 9, no deviation from FCO specifications was observed, confirming the formulation’s stability, non-reactivity, and compatibility with MAP fertilizer. The coating process does not interfere with nutrient concentration, solubility, granule structure, or product performance.[Table 9]>><><

[0055] Transcriptomics study- mode of action- Method for Sample Collection and Transcriptomic Study (RNA-Seq) in Tomato Treated with Biostimulants: To compare transcriptomic changes in tomato plants treated with NUE formulations vs control was performed under field conditions with 100% fertilizer dose and 80% fertilizer dose and identify differentially expressed genes (DEGs) related to NUE, Nutrient uptake etc. Same treatments as outlined in the field design:• T1 - Untreated control- 100% Fertilizer• T2 - Untreated control-80% FertilizerT3 - NUE formulation- 100% FertilizerT4- - NUE formulation- 80% FertilizerSample Collection for Transcriptomics:

[0056] RNA Extraction: Extract total RNA using Qiagen RNeasy Plant Mini Kit or equivalent. Include on-column DNase treatment to remove genomic DNA. Check RNA quality using: Nanodrop (260 / 280 and 260 / 230 ratios), Agilent Bioanalyzer or TapeStation (RIN > 7.5 required) Quantify with Qubit RNA HS Assay.

[0057] Library Preparation and Sequencing: Use poly(A) selection for mRNA enrichment Library prep: RNA Library Prep Kit or equivalent. Index all samples and pool.Perform paired-end sequencing (150 bp) on Illumina NovaSeq or HiSeq platform. Aim for \~30 million reads per sample.

[0058] Bioinformatic Analysis&

[0059] Analysis: Transcriptomic analysis of treated plants revealed a multi-pathway enhancement of NUE. The pathways regulated by the bio formulation of present invention are:&& >&

[0060] According to the embodiment of the present invention, the mode of action of the formulation operates through a multi-tiered mechanistic pathway that enhances nitrogen assimilation, uptake, and utilization within plants as illustrated in [Fig.5] At the primary level, the composition upregulates key enzymatic and metabolic routes associated with nitrogen assimilation, including the glutamine synthetase-glutamate synthase (GS-GOGAT) cycle, nitrate reductase (NR), and nitric oxide-associated pathways, thereby increasing the conversion of available nitrogen into bioactive amino compounds. At the secondary level, the formulation facilitates improved nitrogen uptake by modulating transporter systems such as nitrate transporters (NRTs) and ammonium transporters (AMTs), while concurrently influencing regulatory molecules including melatonin and spermine that support root function and nutrient acquisition.At the tertiary level, the composition enhances nitrogen-use optimization by promoting arginine-based nitrogen storage, regulating branched-chain amino acid (BCAA) partitioning, and strengthening nitrogen salvage and remobilization mechanisms under abiotic or biotic stress. Collectively, these mechanistic features contribute to significantly improved nitrogen-use efficiency and overall plant performance.

[0061] In conclusion, the compositions for enhancing nutrient use efficiency in plants, as disclosed in the present invention present a scientifically validated, scalable, and sustainable advancement in modern agriculture. By harnessing the distinct bioactive properties of Durvillaea potatorum and selected botanical extracts, combined with a water-ethanol cold percolation extraction process, this invention establishes a new standard for plant nutrient efficiency. Unlike conventional fermentation-based formulations, the cold percolation method retains a wider spectrum of bioactive compounds, leading to superior nutrient absorption, enhanced stress tolerance, and long-term agricultural viability. The formulation’s proven ability to reduce fertilizer usage by 20% without affecting crop yield highlights its practical significance and environmental advantages. This innovation serves as a sustainable alternative to synthetic fertilizers, aligning with global agricultural priorities by promoting soil health, minimizing environmental impact, and improving overall crop productivity.

[0062] Novelty and Differentiation of the present invention:• Unique Seaweed Species: Durvillea potatorum Unlike most prior art, which focuses on Ascophyllum nodosum, the present formulation uses Durvillea potatorum, a species known for high secondary metabolite content and distinct bioactive compounds like flavonoids that may offer improved nutrient uptake properties.• Water-Ethanol Cold Percolation Extraction Method: Most prior art emphasizes fermentation, enzymatic hydrolysis, or thermal extraction methods. The present cold percolation method is novel and preserves delicate bioactive compounds that may degrade under heat-based processes.• Synergistic Combination with Botanical Extracts: The combination of Durvillea potatorum with specific botanicals (e.g., Triticum aestivum, Sorghum propinquum, Hordeum vulgare extract or other bioactive plant extracts) is novel and underexplored in the prior art. This combination may provide a broader range of bioactive, leading to enhanced nutrient absorption and fertilizer reduction without compromising yield.• Demonstrated Reduction in Fertilizer Requirement (20%): Few patents provide specific data on reducing fertilizer input without yield loss. The present invention provides reducing fertilizer requirement by 20% with noyield impact which is a significant advantage in demonstrating practical utility.• Sustainability and Environmental Impact: Given the global emphasis on reducing fertilizer use due to environmental concerns (e.g., nitrogen leaching and greenhouse gas emissions), this invention addresses a critical market need for sustainable agriculture practices.

[0063] While considerable emphasis has been placed herein on the components and component parts of the preferred embodiments, it will be appreciated that many embodiments can be made and that many changes can be made in the preferred embodiments without departing from the principles of the disclosure. These and other changes in the preferred embodiment as well as other embodiments of the disclosure will be apparent to those skilled in the art from the disclosure herein, whereby it is to be distinctly understood that the foregoing descriptive matter is to be interpreted merely as illustrative of the disclosure and not as a limitation.

Claims

Claims

1. A bio-composition for enhancing nutrient use efficiency in plants, characterized in thatthe bio composition comprises the extract of seaweed Durvillea at 70% w / v that serves as a source of phenolic and tannin-rich constituents and botanical extracts at 20% w / v that functions as a nitrogen-buffering component; and the extraction is conducted using cold percolation method with water-ethanol solvent at the ratio of 60:40 v / v to maximize recovery of active metabolites; andthe method of preparation of the bio composition comprises the steps of• cleaning seaweed biomass by rinsing freshly harvested seaweed with clean water to remove surface salts, sand, and epiphytes;• cleaning botanical biomass by rinsing freshly harvested botanical material with clean water to remove sand and other impurities;• drying the cleaned biomass under shade in a well- ventilated covered structure that allows diffused sunlight while preventing direct UV exposure, maintaining the temperature below 45 °c until the moisture content is reduced to less than 10% or the biomass becomes crisp and brittle;• pulverizing the dried biomass using a hammer mill and passing the resulting material through a 40-mesh sieve to obtain a fine, uniform powder;• storing the powdered biomass in airtight, light-resistant containers at room temperature;• extracting bioactive compounds from the powdered biomass by cold percolation using a water-ethanol solvent system at a volumetric ratio of approximately 60:40 (v / v), with a solvent-to-biomass ratio of about 10: 1, at ambient temperature for a period ranging from 12 hours to 14 days;• removing the solvent from the extract by rotary evaporation under reduced pressure at a temperature between 40°c and 45 °c;• standardizing the extract to contain not less than 1000 ppm polyphenols and not less than 1000 ppm tannins.

2. The bio composition as claimed in claim 1, wherein the botanical extracts are selected from Salvia indica, Salvia officinalis, Triticum aestivum, Sorghum propinquum, Hordeum vulgare, Trigonella frenum-graecum, Trigonella comiculata, Trigonella caerulea, Medicago sativa, Medicago truncatula, Medicago polymorpha, Emblica officinalis, Phyllanthus amarus, Phyllanthus niruri, Phaseolus vulgaris, Phaseolus lunatus, Phaseolusacutifolius, Ocimum tenuiflorum, Ocimum gratissimum and they contain flavonoids, polyphenols, flavonoids, phenolic acids including gallic acid and syringic acid, tannins.

3. The bio composition as claimed in claim 1, wherein the composition comprises phenolics at 10-50,000 ppm, flavonoids at 10-20,000 ppm, tannins at 500-20,000 ppm, polyamines, and amino acid derivatives.

4. The bio composition as claimed in claim 1, wherein the extraction process preserves thermolabile metabolites, enables significantly higher recovery of phenolic and flavonoid constituents, and minimal co-extraction of alginates, mannitol, and other polysaccharide impurities.

5. The bio composition as claimed in claim 1, wherein the extract of Durvillea contributes to antioxidant activity and facilitate modulation of indole-3-acetic acid (lAA)-related signaling pathways, Melatonin signaling, Spermine signaling and Arginine metabolism for uptake and use efficiency and the botanical extract supports improved nitrogen-use efficiency, enhanced nitrogen storage within plant tissues, and subsequent nitrogen remobilization during key developmental stages.

6. The bio composition as claimed in claim 1, wherein the composition has a shelf life of at least four years when maintained within a temperature range of approximately 5-45 °C.

7. The bio composition as claimed in claim 1, wherein Heavy-metal contaminants, including but not limited to lead, cadmium, arsenic, and mercury, are maintained below detectable limits at typically <0.1 ppm, ensuring regulatory compliance and product safety and the composition is non-toxic to humans, animals, pollinators and aquatic species, rendering it suitable for safe agricultural and environmental use.

8. The bio composition as claimed in claim 1, wherein the combined application with NPK fertilizer increases nitrogen uptake by at least 20%, phosphorus uptake by at least 25%, and potassium uptake by at least 25% compared to 100% recommended dose of NPK fertilizer alone.

9. The bio composition as claimed in claim 1, wherein the composition enhances nitrogen assimilation by upregulating GS- GOGAT cycle and nitrate / ammonium transporters.

10. The bio composition as claimed in claim 1, wherein the formulation enables at least 20% reduction in synthetic fertilizer use without yield penalty.

11. The bio composition as claimed in claim 1, wherein the composition is administered as a foliar spray at a concentration of 3-5 mL per liter of water, enabling rapid uptake through leaf tissues, and alternatively the composition is delivered via fertigation systems at application rates of about 800-1000 mL per acre, providing uniform distribution through irrigation channels and facilitating root- zone assimilation and alternatively the composition is delivered as an additive to conventional granular or liquid NPK fertilizers at inclusion levels ranging from approximately 2% to 5% w / w, thereby enhancing nutrient efficiency and complementing standard fertilization programs.

12. The bio composition as claimed in claim 1, wherein at the primary level, the composition upregulates key enzymatic and metabolic routes associated with nitrogen assimilation, including the glutamine synthetase-glutamate synthase (GS-GOGAT) cycle, nitrate reductase (NR), and nitric oxide-associated pathways, thereby increasing the conversion of available nitrogen into bioactive amino compounds; at the secondary level, the formulation facilitates improved nitrogen uptake by modulating transporter systems such as nitrate transporters (NRTs) and ammonium transporters (AMTs), while concurrently influencing regulatory molecules including melatonin and spermine that support root function and nutrient acquisition and at the tertiary level, the composition enhances nitrogen-use optimization by promoting arginine-based nitrogen storage, regulating branched- chain amino acid (BCAA) partitioning, and strengthening nitrogen salvage and remobilization mechanisms under abiotic or biotic stress.