Patent Application
20250214909
This patent application claims priority benefit of UK Patent Application No: GB2404386.1, entitled “A CARRIER-BASED AGRICULTURAL BIOFERTILIZER COMPOSITION”, filed on 27 Mar. 2024. The entire contents of the patent application are hereby incorporated by reference herein in its entirety.
The present disclosure generally relates to the agricultural fertilizer compositions, more particularly the invention relates to a biofertilizer composition and a method for enhancing soil fertility using a carrier-based agricultural biofertilizer composition.
Agricultural biofertilizers can benefit from ongoing developmental research to further enhance their effectiveness and contribute to sustainable agriculture. Here are some key areas of research that can support the development and optimization of agricultural biofertilizers:
Plant growth-promoting Bacillus (PGPB) are beneficial bacteria that enhance plant growth and crop health. These Bacillus species colonize plant roots, stimulating root development and offering various benefits. They produce enzymes like phosphate solubilizing enzymes, breaking down soil-bound nutrients for plants. Bacillus species also fix atmospheric nitrogen, increasing nutrient availability for crops and improving growth. Certain strains inhibit plant pathogens through antibiotics and antimicrobial compounds, reducing disease reliance on chemical pesticides. Bacillus triggers systemic plant resistance, producing defense-related proteins and compounds against diseases, bolstering plant defenses during stresses like pathogen attacks, drought, and salinity. PGPB help plants tolerate abiotic stresses by producing stress-related enzymes and osmoprotectants. They enhance water and nutrient uptake efficiency, enhancing resilience in harsh conditions. Plant growth-promoting Bacillus enhances soil health by improving structure, nutrient cycling, and organic matter decomposition, boosting fertility and productivity. Such soils, with diverse microbial communities including PGPB, create favorable environments for plant growth, fostering sustainable agriculture. Strategies incorporating PGPB reduce chemical inputs, promote ecological balance, and enhance agricultural sustainability.
Conventional agriculture, reliant on chemical fertilizers, presents numerous drawbacks and limitations that affect both the environment and long-term sustainability. Poorly managed chemical fertilizers contribute to environmental pollution, with excess application leading to nutrient runoff into water bodies, causing water pollution and eutrophication. This depletes oxygen in aquatic ecosystems, harming aquatic life. Unchecked use of chemical fertilizers leads to soil degradation, lacking in practices for soil health improvement. This results in nutrient imbalances, reduced fertility, microorganism loss, and soil structure degradation, lowering crop productivity. Synthetic pesticides, often used in conventional agriculture, harm beneficial organisms, causing biodiversity loss. Reduced biodiversity disrupts ecological balance, affecting pest control and potentially promoting pest resistance. Chemical fertilizer production demands substantial energy from fossil fuels, worsening carbon emissions and climate change. Phosphate and potassium mining for fertilizers deplete finite resources. Prolonged exposure to these chemicals endangers farm workers' health, and residue contamination on crops poses risks to consumers. Conventional agriculture's heavy water use intensifies water scarcity, particularly in water-scarce regions, exacerbating resource depletion due to crop growth demands. To counter these issues and promote sustainability, alternative practices like organic farming, integrated pest management, and organic/bio-based fertilizers are adopted. These approaches minimize environmental harm, support soil health, biodiversity, and reduce reliance on non-renewable resources. They prioritize the long-term viability of agricultural systems.
In the light of the aforementioned discussion, there exists a need for a certain system with novel methodologies that would overcome the above-mentioned disadvantages.
The following presents a simplified summary of the disclosure in order to provide a basic understanding of the reader. This summary is not an extensive overview of the disclosure and it does not identify key/critical elements of the invention or delineate the scope of the invention. Its sole purpose is to present some concepts disclosed herein in a simplified form as a prelude to the more detailed description that is presented later.
An embodiment of the present disclosure is directed towards carrier-based agricultural biofertilizer composition.
An embodiment of the present disclosure is directed towards the carrier-based agricultural biofertilizer composition, includes an isolated Bacillus Cereus strain CGAPGPBBS-048 having the deposit accession number KY495213; an agriculturally acceptable liquid formulation including a stabilizer, that may provide stability and support for at least one of the Bacillus species, thereby enhancing the growth and overall health of the plants.
An objective of the present disclosure is directed towards enabling augmented plant growth and bolstering overall plant health.
An objective of the present disclosure is directed towards endowing plant growth-promoting abilities by proficiently generating phytohormones, notably indoleacetic acid, alongside the secretion of siderophores and other secondary metabolites that confer protection against pathogenic organisms.
An objective of the present disclosure is directed towards the effective production of phytohormones, particularly indoleacetic acid, to heighten plant growth and elevate germination rates.
An objective of the present disclosure is directed towards the capacity to generate phytohormone, specifically indole acetic acid, resulting in enhanced plant germination and growth.
An objective of the present disclosure is directed towards promoting robust plant growth and enhancing overall plant health across diverse agricultural settings.
An objective of the present disclosure is directed towards harnessing the inherent potential of the Bacillus Cereus strain CGAPGPBBS-048 to produce essential phytohormones, particularly emphasizing the significance of indoleacetic acid in stimulating growth processes.
An objective of the present disclosure is directed towards maximizing plant health by leveraging the biofertilizer composition's capacity to secrete siderophores and secondary metabolites, offering effective protection against harmful pathogens that could impede plant growth.
An objective of the present disclosure is directed towards strategically generating indoleacetic acid through controlled application, thereby facilitating robust plant growth, especially during critical germination phases.
An objective of the present disclosure is directed towards offering an environmentally friendly approach to enhancing agricultural practices, empowering farmers to foster healthier plant growth and elevated yields without resorting to synthetic chemicals.
An objective of the present disclosure is directed towards enhancing crop resilience and successful cultivation even in challenging environments, achieved by optimizing indoleacetic acid synthesis and its positive impact on plant adaptation.
An objective of the present disclosure is directed towards providing a sustainable solution for agricultural advancement, reducing reliance on excessive chemical interventions and contributing to soil health preservation and ecological equilibrium.
An objective of the present disclosure is directed towards facilitating resource-efficient farming practices aligned with ecological conservation and long-term agricultural sustainability.
An objective of the present disclosure is directed towards improving the quality of harvested produce, translating into tangible benefits for growers and consumers alike.
An objective of the present disclosure is directed towards addressing the evolving needs of agriculture through innovative solutions, balancing productivity, environmental considerations, and economic viability by leveraging the growth-promoting attributes of the Bacillus strain.
An embodiment of the present disclosure directed towards a method for enhancing soil fertility using of a biofertilizer composition
Furthermore, the objects and advantages of this invention will become apparent from the following description and the accompanying annexed drawings.
In the following, numerous specific details are set forth to provide a thorough description of various embodiments. Certain embodiments may be practiced without these specific details or with some variations in detail. In some instances, certain features are described in less detail so as not to obscure other aspects. The level of detail associated with each of the elements or features should not be construed to qualify the novelty or importance of one feature over the others.
It is to be understood that the present disclosure is not limited in its application to the details of construction and the arrangement of components set forth in the following description or illustrated in the drawings. The present disclosure is capable of other embodiments and of being practiced or of being carried out in various ways. Also, it is to be understood that the phraseology and terminology used herein is for the purpose of description and should not be regarded as limiting.
The use of “including”, “comprising” or “having” and variations thereof herein is meant to encompass the items listed thereafter and equivalents thereof as well as additional items. The terms “a” and “an” herein do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced item. Further, the use of terms “first”, “second”, and “third”, and the like, herein do not denote any order, quantity, or importance, but rather are used to distinguish one element from another.
In another exemplary embodiment, the nutrient medium may include carbon sources selected from the group consisting of sucrose and glucose, whereby the carbon sources may provide optimal conditions for bacterial growth and nutrient assimilation.
In another exemplary embodiment, the nutrient medium may include nitrogen sources selected from the group consisting of ammonium sulfate, urea, and peptone, whereby the nitrogen sources may provide optimal conditions for bacterial growth and nutrient assimilation.
In another exemplary embodiment, the step of fermenting conditions may carried out under controlled conditions including temperature, pH, and oxygen levels, to thereby promote optimal bacterial growth and metabolic activity.
In another exemplary embodiment, the step of systematically applying the meticulously prepared bacterial formulation to soil is performed using sprinkler irrigation, spray application, drone-based application, soil mixing techniques, seed deepening methods, plant root deepening methods, or any combination thereof.
In an exemplary embodiment, the carrier-based agricultural biofertilizer composition may include an isolated Bacillus Cereus strain CGAPGPBBS-048 having the deposit accession number KY495213; an agriculturally acceptable liquid formulation including a stabilizer, that may provide stability and support for at least one of the Bacillus species, thereby enhancing the growth and overall health of the plants.
Indole acetic acid, is a plant hormone that plays a crucial role in various aspects of plant growth and development, including cell elongation, root formation, fruit development, and responses to environmental stimuli. Some Bacillus spp. have the ability to produce IAA, and their interactions with plants can have significant positive effects on plant growth and yield. IAA-producing Bacillus strains are capable of synthesizing IAA through various pathways. They convert tryptophan, an amino acid present in the rhizosphere, into IAA using enzymes such as tryptophan-2-monooxygenase or indole-3-acetamide hydrolase. The synthesized IAA is released into the surrounding environment and can be taken up by plant roots. IAA-producing Bacillus strains enhance nutrient availability in the soil by solubilizing inorganic phosphates and fixing atmospheric nitrogen. It promotes root growth and development. The hormone stimulates cell division and elongation in the root system, resulting in a denser and more extensive root network. Increased root surface area improves nutrient and water uptake, leading to healthier and more vigorous plants. It can induce systemic resistance in plants. They activate plant defense mechanisms, such as the production of pathogenesis-related proteins and phytoalexins, to protect plants from pathogenic microorganisms. This enhanced resistance helps plants combat diseases and reduces the need for chemical pesticides.
The use of IAA-producing Bacillus strains for plant growth promotion offers an environmentally friendly alternative to synthetic fertilizers and pesticides. These bacteria enhance nutrient availability without causing environmental pollution and reduce the reliance on chemical inputs. They also contribute to soil health by improving its structure, nutrient cycling, and overall microbial diversity. IAA-producing Bacillus strains help plants cope with various abiotic stresses, such as drought, salinity, and heavy metal toxicity. The hormone regulates stomatal closure, reducing water loss during drought conditions. It also enhances the synthesis of osmoprotectants and antioxidant enzymes, which protect plants against salt stress and oxidative damage caused by heavy metals. Enhanced nutrient availability, root development, induced resistance, and stress tolerance increase plant yield and productivity. Plants inoculated with indole acetic acid-producing bacteria often exhibit higher biomass accumulation, increased fruit/seed production, and improved quality characteristics.
Alkaline soils and high pH cause many severe impacts on crop yield and quality. While indole acetic acid-producing bacteria have numerous benefits for plant growth and yield, they may face certain challenges when applied in alkaline soil conditions. Alkaline soil conditions can negatively affect the activity and survival of IAA-producing Bacillus strain. These bacteria generally thrive in neutral to slightly acidic soil environments. The high pH of alkaline soil can inhibit their growth, nutrient uptake, and metabolic activities, leading to a decrease in indole acetic acid production.
Alkaline soil conditions can disrupt the metabolic pathways of indole acetic acid synthesis in bacteria. The enzymes involved in tryptophan conversion to indole acetic acid may have reduced activity or be inhibited by the high pH, resulting in lower levels of hormone production. This can limit the beneficial effects of indole acetic acid on plant growth and development. Alkaline soil often has limited availability of certain essential nutrients, such as iron and manganese, due to their reduced solubility. Indole acetic acid-producing bacteria require these nutrients as cofactors for enzyme activity and hormone synthesis. In alkaline soil, these nutrients may be less accessible to both the bacteria and plants, affecting their growth-promoting effects.
Decreased root colonization of IAA-producing Bacillus strains. Indole acetic acid-producing bacteria establish a symbiotic relationship with plant roots, colonizing the rhizosphere and rhizoplane. However, alkaline soil conditions can impede the ability of these bacteria to colonize the roots effectively. The high pH can alter root exudation patterns, reducing the attraction and attachment of the bacteria to the root surface. This can limit the direct interaction between the bacteria and plant roots. Alkaline soil conditions can influence the sensitivity and response of plants to indole acetic acid. Higher soil pH can affect the perception and signaling of the hormone within plant cells, potentially reducing the plant's ability to utilize and respond to indole acetic acid produced by the bacteria. This can weaken the growth-promoting effects of the hormone on plant physiology. Alkaline soil environments often favor certain microbial populations that are adapted to these conditions. These indigenous alkaline-tolerant microorganisms may outcompete indole acetic acid-producing bacteria for resources and niche occupancy. As a result, the establishment and survival of these bacteria in alkaline soil can be compromised.
In an exemplary embodiment, an isolated Bacillus Cereus strain CGAPGPBBS-048 may be derived from the alkaline calcareous soil found in a cowpea farming field located in Yola, Nigeria. This strain may have been incorporated into a biofertilizer formulation designed to significantly enhance seed germination in soils with higher pH levels. The chosen strain demonstrates its efficacy in enhancing plant growth and promoting overall plant health by effectively producing IAA, specifically benefiting crops such as Cowpea (Vigna unguiculata), Groundnut (Arachis hypogaea), and Soybean (Glycine max) during both rainy and dry seasons.
Morphological Characteristics of the Bacillus Cereus strain CGAPGPBBS-048:
Physiological Properties of the Bacillus Cereus strain CGAPGPBBS-048:
In another exemplary embodiment, despite possessing valuable inherent traits, certain beneficial bacterial strains may not naturally inhabit a given field soil or, if present, could be limited in numbers or exhibit reduced activity. This may result in an inability to confer advantageous effects on plants within an unaltered or unenhanced rhizosphere.
In order for a bacterial strain with inherent beneficial traits, such as a plant growth-promoting rhizobacteria (PGPR), for a bacterium to have a positive impact on plant growth, it needs to exhibit competitive advantages and establish effective colonization within the rhizosphere during periods of active plant growth. It's crucial to acknowledge that, in the absence of interventions or enhancements, the probability of any indigenous or naturally occurring PGPR, like the bacterial strain CGAPGPBBS-048, conferring benefits to plant growth is rather limited. This emphasizes the importance of creating a conducive environment to facilitate the successful colonization and effectiveness of PGPR strains.
In another exemplary embodiment, The PGPR strain CGAPGPBBS-048, potentially isolated from the rhizosphere, exhibits a noteworthy trait of producing indole acetic acid under higher pH conditions, thereby contributing to enhanced plant growth. Moreover, the utilization of specialized laboratory cultivation techniques might have been employed to optimize the growth and population density of CGAPGPBBS-048, effectively harnessing its full PGPR potential. These carefully tailored growth conditions are likely identified to bolster the competitive edge of CGAPGPBBS-048 upon application to the rhizosphere, ultimately resulting in a favorable impact on plant growth. Essentially, the meticulous determination of growth parameters conducive to the prosperous colonization of CGAPGPBBS-048 within the rhizosphere has paved the way for its remarkable positive influence on plant growth, a feat unattainable under ordinary circumstances.
In another exemplary embodiment, a biologically pure culture of CGAPGPBBS-048 is cultivated to establish a stock culture, with portions of this culture meticulously preserved in cryogenic vials at a temperature of −80° C. When undertaking production runs, the frozen stock culture serves as the inoculum for a flask containing nutrient-rich broth media, all under carefully controlled and optimized conditions.
In another exemplary embodiment, the bacterial culture may undergo cultivation within a temperature spectrum of 30-32° C. during the rainy season or alternatively, within the range of 48-50° C. during the dry season, in various iterations. This flask culture may then magnify through scaling in a fermenter, maintaining akin growth conditions. This amplification may culminate in augmented population proliferation and an accentuated manifestation of plant growth-promoting attributes inherent to CGAPGPBBS-048, as the culture approaches the early stationary phase. Small quantities drawn from this culture at the early stationary phase are meticulously placed in sterilized plastic bags under aseptic conditions. The final product achieves a minimum concentration of the active ingredient, ensuring a viability of at least 3×108 colony-forming units per mL. Strain CGAPGPBBS-048's fermented culture thrives under optimized conditions to optimize bacterial vitality and preserve plant growth-promoting characteristics. The resultant liquid formulation, securely encased in sterile bags, remains amenable to viable count analyses. Particularly noteworthy, CGAPGPBBS-048 responds distinctively to fluctuations in temperature when subjected to diverse thermal conditions. Examinations into its shelf life underscore its robustness. Even subsequent to an 18-month period of storage at both specified temperatures, the minimal count of bacterial cells steadfastly retains at 2×108, thereby underlining the strain's enduring viability over time.
In another exemplary embodiment, the potential phytotoxic effects of Bacillus Cereus strain CGAPGPBBS-048 were evaluated through a field experiment, where symptoms including tip injury, wilting, vein clearing, necrosis, and epinasty/hyponasty were closely monitored. Intriguingly, no such symptoms were observed either during or following the application of Bacillus Cereus CGAPGPBBS-048 across all three crops during both seasons. These compelling observations suggest the absence of any deleterious impact on the plants, underscoring CGAPGPBBS-048 efficacy as a plant growth-promoting bacterium suitable for commercial utilization under both regular and elevated pH conditions. Moreover, its versatility extends to various temperature conditions within the Nigerian context.
In another exemplary embodiment, the Bacillus Cereus strain CGAPGPBBS-048 may capable of producing indole acetic acid in higher pH conditions by expressing a gene encoding an indole acetic acid-producing enzyme.
In another exemplary embodiment, wherein the Bacillus Cereus strain CGAPGPBBS-048 may be isolated from rhizosphere soil of Yola, Nigeria.
In another exemplary embodiment, wherein the Bacillus Cereus strain CGAPGPBBS-048 may be isolated by a serial dilution technique.
In another exemplary embodiment, the Bacillus Cereus strain CGAPGPBBS-048 may has the ability to stimulate plant growth through the production of phytohormones, including indoleacetic acid, thereby regulating plant growth, improving root development, and enhancing overall plant vigor.
Amplification and 16S rRNA Gene Sequence Analysis: After amplifying the partial 16S rRNA gene, cycle sequencing was performed through MACROGEN in Korea. The resulting amplified product underwent sequencing using the forward sequencing reaction mix. To determine homology, the DNA sequence was searched using the BLAST search engine at the NCBI site (ncbi.nlm.nih.gov) and FASTA (ebi.ac.uk). The FASTA homology search revealed similarity to Bacillus megaterium, and the corresponding strain obtained from NCBI was designated as KY495213.
Process for isolation and cultivation of Bacteria from Soil Sample:
Bacillus Cereus CGAPGPBBS-048
Escherichia coli
170.42 ± 9.39d
Example: 7—Commercial Exploitation (viability and phytotoxicity): The Bacillus strain CGAPGPBBS-048 may demonstrate the ability to produce indole acetic acid in higher pH (alkaline soils) and positively influence the germination and yield of diverse crop varieties. Although some rhizospheric microbes may possess advantageous traits for other crops, the presence of specific bacterial populations in the rhizosphere, without modification or supplementation, is primarily may be determined by the availability of substrates, prevailing environmental conditions (such as soil moisture, pH, and organic matter content), and the competition among different microbial communities. These beneficial microbes may establish colonization on plant roots, enhancing nutrient absorption, synthesizing growth hormones, and providing protection against diseases, thereby promoting overall plant growth and health.
Reference throughout this specification to “one embodiment”, “an embodiment”, or similar language means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present disclosure. Thus, appearances of the phrases “in one embodiment”, “in an embodiment” and similar language throughout this specification may, but do not necessarily, all refer to the same embodiment.
Although the present disclosure has been described in terms of certain preferred embodiments and illustrations thereof, other embodiments and modifications to preferred embodiments may be possible that are within the principles and spirit of the invention. The above descriptions and figures are therefore to be regarded as illustrative and not restrictive.
Thus the scope of the present disclosure is defined by the appended claims and includes both combinations and sub-combinations of the various features described hereinabove as well as variations and modifications thereof, which would occur to persons skilled in the art upon reading the foregoing description.
| Number | Date | Country | Kind |
|---|---|---|---|
| GB2404386.1 | Mar 2024 | GB | national |
