Biofertilizer

Abstract

This disclosure relates to biofertilizers. In particular, this disclosure relates to combinations and compositions of biofertilizers containing endophytic bacteria and mycorrhizae, as well as methods of using them.

Description

Technical Field

[0001] The present disclosure relates to the field of biofertilizers. In particular, the present disclosure relates to combinations and compositions of biofertilizers comprising endophytic bacteria and mycorrhizae, and methods of using the same.

Background Art

[0002] In the practice of agriculture, the use of fertilizers is a common practice to condition the soil and enhance plant health and yield. However, the overuse of chemical fertilizers has caused toxicity in plants, flora and fauna, and has had negative side effects on soil health, the soil microbiome, and the environment. As a result, the use of biofertilizers is being promoted to overcome the negative side effects of chemical supplements.

[0003] Biofertilizers are living microorganisms that enhance plant nutrition either by mobilizing or increasing the availability of nutrients in the soil. Various microbial taxa, including beneficial bacteria and fungi, are currently used as biofertilizers because they successfully colonize the rhizosphere, rhizoplane or inside the roots and promote growth by increasing the supply or availability of primary nutrients to the host plant. Arbuscular mycorrhizae (or "mycorrhizae") are beneficial fungi that create a transport environment for nutrient uptake in the plant root zone. Mycorrhizae are a symbiotic relationship between plant roots and fungi. The main role of mycorrhizae is to enhance the uptake of nutrients and water by the host plant by accessing a greater volume of soil than the roots alone can utilize. Mycorrhizae occur in many forms and depend on both the host plant and the fungal taxonomic system. Mycorrhizae play an important role in plant nutrition, soil biology, and the chemical properties of the soil. Currently, several mycorrhiza-based biofertilizers have attracted attention for improving soil health, plant health and yield.

[0004] Despite the availability of numerous biofertilizers, research into the optimal combination of mycorrhizal elements possessing diverse properties that can have the greatest impact on soil and plant health remains insufficient. Biofertilizer compositions containing mycorrhizae, with enhanced beneficial functions, are still needed. Compositions that enhance and optimize mycorrhizal function, and thus maximize benefits for soil and crops, are required. Biofertilizer compositions that promote and rejuvenate soil health and the soil microbiome are also needed.

[0005] In accordance with the needs again targeted herein, the object of this disclosure is to provide combinations or compositions containing mycorrhizae with enhanced mycorrhizal function. The object of this disclosure is to add agents that enhance mycorrhizal properties to compositions containing mycorrhizae. The object of this disclosure is to provide combinations or compositions containing mycorrhizae and endophytic bacteria. The object of this disclosure is to improve plant health, yield and soil properties using combinations or compositions containing mycorrhizae and endophytic bacteria. [Overview of the project]

[0006] One aspect of the present disclosure provides a combination of biofertilizers comprising endophytic bacteria selected from Pseudomonas fluorescens, Bacillus, Bacillus amyloliquefaciens, Pseudomonas putida, Paenibacillus polymyxa, or a mixture thereof, and one or more mycorrhizae.

[0007] One aspect of the present disclosure provides a combination of biofertilizers comprising an endophytic bacterium selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, and one or more mycorrhizae.

[0008] In one embodiment, the mycorrhizae may be selected from endomycorrhizae, ectomycorrhizae, or a combination thereof. In another embodiment, the mycorrhizae may be in the form of vegetative growths selected from endomycorrhizal vegetative growths, ectomycorrhizal vegetative growths, or a combination thereof.

[0009] Another aspect of the present disclosure provides a combination of biofertilizers comprising endophytic bacteria selected from Pseudomonas fluorescens, Bacillus, Bacillus amyloliquefaciens, Pseudomonas putida, Paenibacillus polymyxa, or a mixture thereof, and one or more mycorrhizae.

[0010] Another aspect of the present disclosure provides a biofertilizer composition comprising an endophytic bacterium selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, and one or more mycorrhizae.

[0011] One aspect of the present disclosure provides a method for preparing a biofertilizer composition comprising endophytic bacteria selected from Pseudomonas fluorescens, Bacillus subtilis, Bacillus amyloliquefaciens, Pseudomonas putida, Paenibacillus polymyxa, or a mixture thereof, and one or more mycorrhizae.

[0012] One aspect of the present disclosure provides a method for preparing a biofertilizer composition comprising an endophytic bacterium selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, and one or more mycorrhizae.

[0013] One aspect of the present disclosure provides a method for improving the health, yield and vitality of a plant, the method comprising contacting a plant or a part thereof, or a growing area (locus), with a combination of biofertilizers comprising endophytic bacteria selected from Pseudomonas fluorescens, Bacillus subtilis, Bacillus amyloliquefaciens, Pseudomonas putida, Paenibacillus polymyxa, or a mixture thereof, and one or more mycorrhizae.

[0014] One aspect of the present disclosure provides a method for improving the health, yield and vitality of a plant, the method comprising contacting a plant or a part thereof, or a growing area, with a combination of biofertilizers comprising endophytic bacteria selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, and one or more mycorrhizae.

[0015] Another aspect of the present disclosure provides the use of a biofertilizer composition comprising endophytic bacteria selected from Pseudomonas fluorescens, Bacillus subtilis, Bacillus amyloliquefaciens, Pseudomonas putida, Paenibacillus polymyxa, or a mixture thereof, and one or more mycorrhizae, for improving plant health, yield, and vitality.

[0016] Another aspect of the present disclosure provides the use of a combination of biofertilizers for improving plant health, yield, and vitality, comprising endophytic bacteria selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, and one or more mycorrhizae.

[0017] One aspect of the present disclosure also provides a kit comprising a component comprising endophytic bacteria selected from Pseudomonas fluorescens, Bacillus subtilis, Bacillus amyloliquefaciens, Pseudomonas putida, Paenibacillus polymyxa, or a mixture thereof, and another component comprising one or more mycorrhizae for improving plant health, yield, and vitality.

[0018] One aspect of the present disclosure also provides a kit comprising a component comprising an endophytic bacterium selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, and another component comprising one or more mycorrhizae for improving plant health, yield, and vitality.

[0019] In one embodiment, the kit may be divided into multiple components of endophytic bacteria and one or more mycorrhizae, which are mixed together, applied together, together, sequentially, or simultaneously. [Modes for carrying out the invention]

[0020] For the purposes of the following detailed description, it should be understood that the present invention may envision various alternative modifications and step sequences, unless otherwise explicitly specified. Furthermore, unless otherwise stated, except in any example of operation, all numbers representing quantities of materials / components used herein should be understood in all cases to be modified by the term "about".

[0021] Therefore, before describing the present invention in detail, it should be understood that the present invention is not limited to particularly exemplified process parameters or compositions, which are naturally subject to change. It should also be understood that the terms used herein are for the purpose of illustrating embodiments of the present invention and are not intended to limit the scope of the present invention in any way. The use of examples anywhere in this specification, including examples of any terms discussed herein, is illustrative only and does not limit the scope and meaning of the present invention or any exemplified terms. Similarly, the present invention is not limited to the various embodiments given herein. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present invention pertains. In case of any conflict, the definitions in this document shall prevail.

[0022] It should also be noted that, as used herein, the singular forms "a," "an," and "the" refer to multiple subjects unless the content explicitly indicates otherwise. The terms "preferred" and "preferably" refer to embodiments of the invention that may provide particular benefits under certain circumstances.

[0023] As used herein, “comprising,” “including,” “having,” “containing,” “involving,” and similar terms should be understood to mean unrestricted, i.e., including without limitation.

[0024] Unless otherwise specified, all exact values provided in this specification represent corresponding approximate values (e.g., all exact exemplary values provided for a particular factor or measured value can be considered to also provide the corresponding approximate measured value modified by "about" where appropriate). As used herein, the term "about" refers to measurable values such as parameters, amounts, time, etc., and includes variations of up to ±15% or less, specifically ±10% or less, more specifically ±5% or less, even more specifically ±1% or less, and still more specifically ±0.1% or less from a particular recited value, as long as such variations are appropriate in the practice of the disclosure described herein.

[0025] The recitation of a range of values is intended to be merely a shorthand way of referring individually to each separate value falling within that range, unless otherwise indicated herein, and each separate value is incorporated herein as if it were individually recited herein. All ranges of values provided are intended to include the endpoints and the values between the endpoints and are combinable independently. As used herein, all numerical values or ranges of numerical values include the integers within such range and the fractional part of such value, or the integers within such range, unless the context clearly indicates otherwise. Thus, for example, reference to a range of 90% to 100% includes 91%, 92%, 93%, 94%, 95%, 96%, 97%, etc., as well as 91.1%, 91.2%, 91.3%, 91.4%, 91.5%, etc., 92.1%, 92.2%, 92.3%, 92.4%, 92.5%, etc. All methods described herein can be performed in the appropriate order, unless otherwise indicated herein or clearly inconsistent with the context.

[0026] With the aim in mind, the inventors of the present disclosure surprisingly found that by combining endophytic bacteria with mycorrhizae, the functions of mycorrhizae are enhanced and have a greater impact on soil and plant health. More specifically, the inventors of the present application found that bacteria such as Pseudomonas, Bacillus, and Paenibacillus significantly enhance the functions of mycorrhizae and act symbiotically to bring about significant effects in the plants to which they are applied. Furthermore, the inventors of the present application surprisingly observed that the presence of endophytic bacteria combined with mycorrhizae increases the colonization by mycorrhizal fungi.

[0027] Thus, one embodiment of the present disclosure provides a combination of biofertilizers comprising endophytic bacteria selected from Pseudomonas fluorescens, Bacillus subtilis, Bacillus amyloliquefaciens, Pseudomonas putida, Paenibacillus polymyxa, or a mixture thereof, and one or more mycorrhizae.

[0028] In one embodiment, Pseudomonas fluorescens is the one deposited as MTCC 25519.

[0029] In one embodiment, Bacillus subtilis is the one deposited as MTCC 25520.

[0030] In one embodiment, Bacillus amyloliquefaciens is the one deposited as MTCC 25518.

[0031] In one embodiment, Pseudomonas putida is the one deposited as MTCC 25607.

[0032] In one embodiment, Paenibacillus polymyxa is the one deposited as MTCC 25609.

[0033] Accordingly, one embodiment of the present disclosure provides a combination of biofertilizers comprising an endophytic bacterium selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, and one or more mycorrhizae.

[0034] One embodiment of the present disclosure provides a biofertilizer composition comprising a combination of the biofertilizers of the present disclosure.

[0035] Accordingly, one embodiment of the present disclosure provides a biofertilizer composition comprising an endophytic bacterium selected from Pseudomonas fluorescens, Bacillus, Bacillus amyloliquefaciens, Pseudomonas putida, Paenibacillus polymyxa, or a mixture thereof, and one or more mycorrhizae.

[0036] Accordingly, one embodiment of the present disclosure provides a biofertilizer composition comprising an endophytic bacterium selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, and one or more mycorrhizae.

[0037] In one embodiment, the above combination includes at least one type of endophytic bacteria and one or more types of mycorrhizae. In one embodiment, the above combination includes at least two types of endophytic bacteria and one or more types of mycorrhizae. In one embodiment, the above combination includes at least three types of endophytic bacteria and one or more types of mycorrhizae. In one embodiment, the above combination includes at least four types of endophytic bacteria and one or more types of mycorrhizae. In one embodiment, the above combination includes five types of endophytic bacteria and one or more types of mycorrhizae.

[0038] In a preferred embodiment, the endophytic bacteria are a mixture of Pseudomonas fluorescens deposited as MTCC 25519 and Bacillus subtilis deposited as MTCC 25520.

[0039] Another embodiment of the present disclosure provides a combination of biofertilizers comprising Pseudomonas fluorescens deposited as MTCC 25519 and one or more mycorrhizae.

[0040] Another embodiment of the present disclosure provides a combination of biofertilizers comprising Bacillus subtilis deposited as MTCC 25520 and one or more mycorrhizae.

[0041] Another embodiment of the present disclosure provides a combination of biofertilizers comprising Bacillus amyloliquefaciens deposited as MTCC 25518 and one or more mycorrhizae.

[0042] Another embodiment of the present disclosure provides a combination of biofertilizers comprising Pseudomonas putida deposited as MTCC 25607 and one or more mycorrhizae.

[0043] Another embodiment of the present disclosure provides a combination of biofertilizers comprising Paenibacillus polymyxa deposited as MTCC 25609 and one or more mycorrhizae.

[0044] In one embodiment, the mycorrhizae are endomycorrhizae, ectomycorrhizae, or a combination thereof. Entomycorrhizae and ectomycorrhizae are biologically symbiotic soil fungi that form colonies on plant root systems, acting as living extensions of the roots, increasing the absorption surface area, and improving plant nutrient and water efficiency. Through this symbiosis, endomycorrhizae and / or ectomycorrhizae provide the host plant with direct and indirect benefits, including, but not limited to, increased stress resilience, drought tolerance, and reduced fertilizer and irrigation requirements.

[0045] In one embodiment, the mycorrhizae include mycorrhizal vegetative growths. In one embodiment, the mycorrhizal vegetative growths are selected from endomycorrhizal vegetative growths, ectomycorrhizal vegetative growths, or a combination thereof.

[0046] In one embodiment, the mycorrhizal vegetative growth includes vegetative growth of endomycorrhizae selected from arbuscular mycorrhizae, ericoid mycorrhizae, arbutoid mycorrhizae, monotropoid mycorrhizae, orchid mycorrhizae, or combinations thereof.

[0047] In one embodiment, the mycorrhizal vegetative propagule includes an endomycorrhizal vegetative propagule selected from the class Glomeromycota (Glomusmycota). In one embodiment, the mycorrhizal vegetative propagule of the class Glomeromycota is selected from the orders Archaeosporales, Diversisporales, Glomerales, Paraglomerales, or a combination thereof.

[0048] In one embodiment, the mycorrhizal vegetative body is composed of an endomycorrhizal vegetative body, which includes vegetative bodies of Glomaceae (Glomeraceae), Acaulosporaceae, Archaeosporaceae, Diversisporaceae, Pacisporaceae, Paragalomaceae, Gigasporaceae, Ambisporaceae, Geosiphonaceae, Claroideoglomeraceae, or combinations thereof. In one embodiment, the mycorrhizal vegetative body is composed of an endomycorrhizal vegetative body, which includes vegetative bodies of the genus Glomus, Entrophospora, Funneliformis, Rhizophagus, Septoglomus, Acaulospora, Archaeosporaceae, Paragalomaceae, Gigasporaceae, Pacispora, Sclerocystis, Dentiscutata, Scutellospora, and combinations thereof.

[0049] In one embodiment, the mycorrhizal vegetative growth includes vegetative growth of an endomycorrhizae selected from the group consisting of Rhizophagus irregularis, Rhizophagus clarus, Gigaspora margarita, Scutellospora heterogama, Glomus mossae, Septoglomus viscosum, Funneliformis mossae, and combinations thereof.

[0050] As used herein, the term "Scutellospora heterogama" also refers to Dentiscutata heterogama and its subspecies and strains.

[0051] In one embodiment, the mycorrhizal vegetative growth includes vegetative growth of an endomycorrhizal selected from the group consisting of Rhizophagus irregularis, Rhizophagus clarus, Gigaspora margarita, Scutellospora heterogama, and combinations thereof.

[0052] In one embodiment, the mycorrhizal vegetative growth includes a combination of endomycorrhizal vegetative growths. In some embodiments, the combination of the present disclosure is (i) vegetative propagates of Rhizophagus irregularis and Rhizophagus clarus, (ii) vegetative propagates of Rhizophagus irregularis and Gigaspora margarita, (iii) vegetative propagates of Rhizophagus irregularis and Scutellospora heterogama, (iv) vegetative propagates of Rhizophagus clarus and Gigaspora margarita, (v) vegetative propagates of Rhizophagus clarus and Scutellospora heterogama, (vi) vegetative propagates of Gigaspora margarita and Scutellospora heterogama, (vii) vegetative propagates of Rhizophagus irregularis, Rhizophagus clarus, and Gigaspora margarita, (viii) vegetative propagates of Rhizophagus irregularis, Rhizophagus clarus, and Scutellospora heterogama, (ix) Rhizophagus irregularis, Gigaspora This includes vegetative propagates of (x)Rhizophagus irregularis, Rhizophagus clarus, Gigaspora margarita, and Scutellospora heterogama, as well as combinations of endomycorrhizal vegetative propagates selected from vegetative propagates of (x)Rhizophagus irregularis, Rhizophagus clarus, Gigaspora margarita, and Scutellospora heterogama.

[0053] In preferred embodiments, the combination of mycorrhizal vegetative organisms includes a combination of Rhizophagus irregularis, Rhizophagus clarus, Gigaspora margarita, and Scutellospora heterogama.

[0054] In one embodiment, the mycorrhizal vegetative growth includes ectomycorrhizal vegetative growths selected from Laccaria, Pisolithus, Piriformospora, Scleroderma, Rhizopogon, and combinations thereof.

[0055] In one embodiment, the mycorrhizal vegetative propagates include, but are not limited to, ectomycorrhizal vegetative propagates selected from Piriformospora indica, Laccaria bicolor, Laccaria lacata, Rhizopogon roseolus, Laccaria proxima, Pisolithus arhizus, Pisolithus tinctorius, and combinations thereof.

[0056] In one embodiment, the mycorrhizal vegetative propagule includes a combination of ectomycorrhizal vegetative propagules. In one embodiment, the combination of mycorrhizal vegetative propagules includes a combination of Piriformospora indica, Laccaria bicolor, and Pisolithus tinctorius. In one embodiment, the combination of mycorrhizal vegetative propagules includes a combination of Piriformospora indica, Laccaria bicolor, Laccaria lacata, and Pisolithus tinctorius.

[0057] In some embodiments, the compositions of the present disclosure include: (i) vegetative propagates of Piriformospora indica and Laccaria bicolor; (ii) vegetative propagates of Piriformospora indica and Pisolithus tinctorius; (iii) vegetative propagates of Laccaria bicolor and Pisolithus tinctorius; (iv) vegetative propagates of Piriformospora indica, Laccaria bicolor, and Pisolithus tinctorius; (v) vegetative propagates of Piriformospora indica and Laccaria lacata; (vi) vegetative propagates of Laccaria lacata and Pisolithus tinctorius; (vii) vegetative propagates of Laccaria lacata and Laccaria bicolor; (viii) vegetative propagates of Piriformospora indica, Laccaria lacata, and Pisolithus This includes vegetative propagates of (ix) Piriformospora indica, Laccaria bicolor, and Laccaria lacata, (x) vegetative propagates of Laccaria bicolor, Laccaria lacata, and Pisolithus tinctorius, and combinations of ectomycorrhizal vegetative propagates selected from (xi) Piriformospora indica, Laccaria bicolor, Laccaria lacata, and Pisolithus tinctorius.

[0058] In one embodiment, the mycorrhizal vegetative growth includes a combination of endomycorrhizae and ectomycorrhizae. The endomycorrhizae and ectomycorrhizae may be selected from the species described above.

[0059] In one embodiment, endophytic bacteria and mycorrhizae are mixed in a ratio of 1:100 to 100:1. In one embodiment, endophytic bacteria and mycorrhizae are mixed in a ratio of 1:75 to 75:1. In one embodiment, endophytic bacteria and mycorrhizae are mixed in a ratio of 1:50 to 50:1. In one embodiment, endophytic bacteria and mycorrhizae are mixed in a ratio of 1:30 to 30:1. In one embodiment, endophytic bacteria and mycorrhizae are mixed in a ratio of 1:20 to 20:1. In a preferred embodiment, endophytic bacteria and mycorrhizae are mixed in a ratio of 1:17 to 2:1.

[0060] The term “mixed” also refers to synonyms such as “combined” and “mixed” in the context of the embodiments provided herein.

[0061] In one embodiment, the endophytic bacteria are a mixture of Pseudomonas fluorescens deposited as MTCC 25519 and Bacillus subtilis deposited as MTCC 25520, where Pseudomonas fluorescens deposited as MTCC 25519 and Bacillus subtilis deposited as MTCC 25520 are mixed in a ratio ranging from 1:100 to 100:1, preferably 1:75 to 75:1, more preferably 1:50 to 50:1, and even more preferably 1:30 to 30:1. In one embodiment, a preferred ratio is 1:20 to 20:1.

[0062] In one embodiment, endophytic bacteria are present in an amount ranging from about 0.1% to about 50% by weight of the total composition. In one embodiment, endophytic bacteria are present in an amount ranging from about 0.1% to about 30% by weight of the total composition. In one embodiment, endophytic bacteria are present in an amount ranging from about 0.1% to about 20% by weight of the total composition. In one embodiment, endophytic bacteria are present in an amount ranging from about 0.5% to about 10% by weight of the total composition. In one embodiment, endophytic bacteria are present in an amount ranging from about 2% to about 10% by weight of the total composition.

[0063] The biofertilizer compositions containing the above-mentioned combinations include endomycorrhizae, ectomycorrhizae, or combinations thereof in an amount ranging from approximately 0.1% to approximately 90% by weight of the composition.

[0064] The composition contains endomycorrhizae in an amount ranging from about 0.1% to about 90% by weight of the composition. In one embodiment, the mycorrhizal vegetative propagule comprises endomycorrhizal vegetative propagules selected from one or more of Rhizophagus irregularis, Rhizophagus clarus, Gigaspora margarita, and Scutellospora heterogama. In one embodiment, the mycorrhizal vegetative propagule comprises a combination of endomycorrhizal vegetative propagules. As mentioned above, the combination of mycorrhizal vegetative propagules includes various combinations of Rhizophagus irregularis, Rhizophagus clarus, Gigaspora margarita, and Scutellospora heterogama.

[0065] In one embodiment, the combination of biofertilizers and the composition containing the above-mentioned combination contain a combination of endomycorrhizae and ectomycorrhizae in a weight ratio in the range of 1:10 to 10:1. In one embodiment, the weight ratio of endomycorrhizae to ectomycorrhizae in the combination of biofertilizers and the composition containing the above-mentioned combination is in the range of 1:9 to 9:1. In one embodiment, the weight ratio of endomycorrhizae to ectomycorrhizae in the combination of biofertilizers and the composition containing the above-mentioned combination is in the range of 3:7 to 7:3. In one embodiment, the weight ratio of endomycorrhizae to ectomycorrhizae in the combination of biofertilizers and the composition containing the above-mentioned combination is in the range of 2:3 to 3:2. In one embodiment, the weight ratio of endomycorrhizae to ectomycorrhizae in the combination of biofertilizers and the composition containing the above-mentioned combination is in the range of 1:5 to 5:1, preferably 1:4 to 4:1.

[0066] In one embodiment, the mycorrhizal vegetative growth includes a combination of endomycorrhizae and ectomycorrhizae, with the endomycorrhizal vegetative growth present in an amount ranging from approximately 10% to approximately 80% by weight of the mycorrhizal vegetative growth.

[0067] In one embodiment, the mycorrhizal vegetative growth includes a combination of endomycorrhizae and ectomycorrhizae, and the ectomycorrhizal vegetative growth is present in an amount ranging from approximately 20% to approximately 90% by weight of the mycorrhizal vegetative growth.

[0068] In one embodiment, the mycorrhizal vegetative growth comprises (a) an amount of endomycorrhizal vegetative growth selected from Rhizophagus irregularis, Rhizophagus clarus, Gigaspora margarita, Scutellospora heterogama and combinations thereof, in the range of about 10% to about 80% by weight of the mycorrhizal vegetative growth; and (b) an amount of ectomycorrhizal vegetative growth selected from Piriformospora indica, Laccaria bicolor, Laccaria lacata, Pisolithus tinctorius and combinations thereof, in the range of about 20% to about 90% by weight of the mycorrhizal vegetative growth.

[0069] In one embodiment, the mycorrhizal vegetative growth comprises (a) an endomycorrhizal vegetative growth selected from Rhizophagus irregularis, Rhizophagus clarus, Gigaspora margarita, Scutellospora heterogama and combinations thereof, in an amount of about 70% by weight of the mycorrhizal vegetative growth; and (b) an ectomycorrhizal vegetative growth selected from Piriformospora indica, Laccaria bicolor, Laccaria lacata, Pisolithus tinctorius and combinations thereof, in an amount of about 30% by weight of the mycorrhizal vegetative growth.

[0070] In one embodiment, the mycorrhizal vegetative growth comprises (a) an amount in the range of about 30% by weight of the mycorrhizal vegetative growth of endomycorrhizal growth selected from Rhizophagus irregularis, Rhizophagus clarus, Gigaspora margarita, Scutellospora heterogama and combinations thereof, and (b) an amount in the range of about 70% by weight of the mycorrhizal vegetative growth of ectomycorrhizal growth selected from Piriformospora indica, Laccaria bicolor, Laccaria, lacata, Pisolithus tinctorius and combinations thereof.

[0071] In one embodiment, the number of vegetative propagators in the combination or composition of the Disclosure is about 1 × 10 2 Individual vegetatively propagated organisms / g ~ approximately 9 x 10 6 The range is one vegetative propagator / g. In one embodiment, the combination or composition of the present disclosure is about 2 × 10 2 ~2×10 5 Contains vegetative propagators / g. In one embodiment, the combination or composition of the present disclosure is about 1 × 10 5 Contains vegetative propagators / g. In one embodiment, the combination or composition of the present disclosure is about 2 × 10 2 Contains individual vegetative reproductive organisms / g.

[0072] In one embodiment, the composition of the present disclosure further comprises one or more agriculturally acceptable excipients. In one embodiment, the agriculturally acceptable excipients are selected from any or a combination of wetting agents, dispersants, binders, surfactants / emulsifiers, antifreezes, minerals, fillers, stabilizers, and preservatives. However, it should be understood that any other agriculturally acceptable excipients, as known to those skilled in the art, may be used to achieve the intended purpose without departing from the scope of the present disclosure. In one embodiment, the agriculturally acceptable excipients are present in an amount ranging from 0.3% to 99% by weight of the composition.

[0073] Accordingly, one embodiment of the present disclosure provides a biofertilizer composition comprising a combination of the biofertilizers of the present disclosure and an agriculturally acceptable excipient.

[0074] Accordingly, one embodiment of the present disclosure provides a biofertilizer composition comprising an endophytic bacterium selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, one or more mycorrhizae, and an agriculturally acceptable excipient.

[0075] In one embodiment, the agriculturally acceptable excipient may be a stabilizer selected from any or a combination of humic acid, cellulose, dextrose, lactose, methylcellulose, xanthan gum, polyvinylpyrrolidone (PVP), Tween, and gum arabic. In one embodiment, the stabilizer is present in the composition in an amount ranging from about 0.2% to about 80% by weight of the composition.

[0076] One embodiment provides a biofertilizer composition comprising an endophytic bacterium selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, one or more mycorrhizae, a stabilizer, and an agriculturally acceptable excipient. In one embodiment, the agriculturally acceptable excipient is selected from the group including, but is not limited to, wetting agents, dispersants, binders, surfactants / emulsifiers, antifreezes, minerals, fillers, preservatives, and combinations thereof.

[0077] The wetting agent is selected from the group including, but is not limited to, sulfosuccinates, naphthalene sulfonates, sulfated esters, phosphate esters, sulfated alcohols, alkylbenzene sulfonates, and combinations thereof.

[0078] The dispersant is selected from the group including, but is not limited to, polycarboxylates, naphthalene sulfonate condensates, phenol sulfonic acid condensates, lignosulfonates, methyl oleyl taurates, polyvinyl alcohols, and combinations thereof.

[0079] The binder is selected from the group including, but is not limited to, polyvinyl alcohol, phenylnaphthalene sulfonate, lignin derivatives, polyvinylpyrrolidone, polyalkylpyrrolidone, carboxymethylcellulose, xanthan gum, polyethoxylated fatty acids, polyethoxylated fatty alcohols, ethylene oxide copolymer, propylene oxide copolymer, polyethylene glycol, polyethylene oxide, and combinations thereof.

[0080] Surfactants are selected from the group including ionic surfactants, nonionic surfactants, and combinations thereof. Non-limiting examples of ionic surfactants include sulfonic acids, sulfate esters, carboxylic acids, and salts thereof. Non-limiting examples of water-soluble anionic surfactants include alkyl sulfates, alkyl ether sulfates, alkylamide ether sulfates, alkylaryl polyether sulfates, alkylaryl sulfates, alkylaryl sulfonates, monoglyceride sulfates, alkyl sulfonates, alkylamide sulfonates, alkylaryl sulfonates, benzene sulfonates, toluene sulfonates, xylene sulfonates, cumene sulfonates, alkylbenzene sulfonates, alkyldiphenyl oxide sulfonates, alpha-olefin sulfonates, alkylnaphthalene sulfonates, paraffin sulfonates, lignin sulfonates, alkyl sulfosuccinates, ethoxylated sulfosuccinates, alkyl ether sulfosuccinates, alkylamide sulfosuccinates, alkyl sulfosuccinates, alkyl sulfoacetates, alkyl phosphates, phosphate esters, alkyl ether phosphates, acyl sarcosinates, acyl isethionates, N-acyl taurates, N-acyl-N-alkyl taurates, and alkyl carboxylates.

[0081] Non-limiting examples of nonionic surfactants include glycerol ethers, glycol ethers, ethanolamides, sulfoanylamides, alcohols, amides, alcohol ethoxylates, glycerol esters, glycol esters, ethoxylates of glycerol esters and glycol esters, sugar-based alkyl polyglycosides, polyoxyethylene-modified fatty acids, alkanolamine condensates, alkanolamides, tertiary acetylene glycols, polyoxyethylene-modified mercaptans, carboxylic acid esters, polyoxyethylene-modified polyoxypropylene glycols, sorbitan fatty acid esters, or combinations thereof. Also included are EO / PO block copolymers (where EO is ethylene oxide and PO is propylene oxide), EO polymers and copolymers, polyamines, and polyvinylpyrrolidone, sorbitan fatty acid alcohol ethoxylates and sorbitan fatty acid ester ethoxylates.

[0082] The antifreeze is selected from the group including, but is not limited to, ethylene glycol, propylene glycol, urea, glycerin, antifreeze proteins, and combinations thereof.

[0083] The minerals are selected from the group including, but are not limited to, kaolin, silica, titanium(IV) oxide, rutile, anatase, aluminum oxide, aluminum hydroxide, iron oxide, iron sulfide, magnetite, pyrite, hematite, ferrite, gleigate, calcium carbonate, calcite, aragonite, quartz, zircon, olivine, orthopyroxene, tourmaline, kyanite, albite, anorthite, clinopyroxene, orthoclase, gypsum, andalusite, talc, fluorite, apatite, orthoclase, topaz, corundum, diamond, tin, tin oxide, antimony, antimony oxide, beryllium, cobalt, copper, feldspar, gallium, indium, lead, lithium, manganese, mica, molybdenum, nickel, pearlite, platinum group metals, phosphorus and phosphate rock, potash, rare earth elements, tantalum, tungsten, vanadium, zeolite, zinc and zinc oxide, indium tin oxide, and combinations thereof.

[0084] The filler is selected from the group including, but is not limited to, diatomaceous earth, kaolin, bentonite, precipitated silica, attapulgite, perlite, and combinations thereof.

[0085] One embodiment of the present disclosure provides a biofertilizer composition comprising a combination of biofertilizers of the present disclosure and a plant extract.

[0086] One embodiment provides a biofertilizer composition comprising an endophytic bacterium selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, one or more mycorrhizae, and a plant extract.

[0087] In one embodiment, the plant extract acts as a gelling agent or binder. In one embodiment, the plant extract is derived from a plant belonging to the genus Dodonaea. In one embodiment, the plant extract is derived from a plant belonging to the genus Dodonaea viscosa.

[0088] In one embodiment, the extract of a plant belonging to the genus Dodonaea is an aqueous extract.

[0089] In one embodiment, an extract from a plant belonging to the genus Dodonaea is used as the solvent.

[0090] In one embodiment, the composition contains an extract of a plant belonging to the genus Dodonaea in an amount ranging from about 0.1% to about 40% by weight of the composition. In one embodiment, the composition contains an extract of a plant belonging to the genus Dodonaea in an amount ranging from about 0.3% to about 30% by weight of the composition. In one embodiment, the composition contains an extract of a plant belonging to the genus Dodonaea in an amount ranging from about 0.5% to about 20% by weight of the composition. In one embodiment, the composition contains an extract of a plant belonging to the genus Dodonaea in amounts of approximately 0.1% to approximately 30% by weight, approximately 0.1% to approximately 25% by weight, approximately 0.1% to approximately 20% by weight, approximately 0.1% to approximately 15% by weight, approximately 0.1% to approximately 10% by weight, approximately 0.3% to approximately 25% by weight, approximately 0.3% to approximately 20% by weight, approximately 0.3% to approximately 15% by weight, approximately 0.3% to approximately 10% by weight, approximately 0.5% to approximately 40% by weight, approximately 0.5% to approximately 30% by weight, approximately 0.5% to approximately 25% by weight, approximately 0.5% to approximately 15% by weight, approximately 0.5% to approximately 10% by weight, approximately 1% to approximately 40% by weight, approximately 1% to approximately 30% by weight, approximately 1% to approximately 1% by weight. Contains in amounts ranging from approximately 25% by weight, approximately 1% to approximately 20% by weight, approximately 1% to approximately 15% by weight, approximately 1% to approximately 10% by weight, approximately 2% to approximately 40% by weight, approximately 2% to approximately 35% by weight, approximately 2% to approximately 30% by weight, approximately 2% to approximately 25% by weight, approximately 2% to approximately 20% by weight, approximately 2% to approximately 15% by weight, approximately 2% to approximately 10% by weight, approximately 5% to approximately 40% by weight, approximately 5% to approximately 35% by weight, approximately 5% to approximately 30% by weight, approximately 5% to approximately 20% by weight, approximately 5% to approximately 15% by weight, approximately 10% to approximately 40% by weight, approximately 10% to approximately 35% by weight, approximately 10% to approximately 30% by weight, approximately 15% to approximately 30% by weight, or approximately 20% to approximately 40% by weight.

[0091] One embodiment provides a biofertilizer composition comprising an endophytic bacterium selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, one or more mycorrhizae, a plant extract, and an agriculturally acceptable excipient.

[0092] One embodiment provides a biofertilizer composition comprising an endophytic bacterium selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, one or more mycorrhizae, a plant extract, a stabilizer, and an agriculturally acceptable excipient. In one embodiment, the agriculturally acceptable excipient may be a stabilizer selected from humic acid, cellulose, dextrose, lactose, methylcellulose, xanthan gum, polyvinylpyrrolidone (PVP), Tween, and gum arabic, or a combination thereof. In one embodiment, agriculturally acceptable excipients are selected from the group including, but are not limited, wetting agents, dispersants, binders, surfactants / emulsifiers, antifreezes, minerals, fillers, preservatives, and combinations thereof.

[0093] The advantageous agricultural compositions of this disclosure can be formulated into any of the following formulations: aerosols, emulsifying concentrates, wettable powders, soluble concentrates, soluble powders, suspension concentrates, spray concentrates, capsule suspensions, water-dispersible granules, granules, dust, or fine granular seed treatment formulations.

[0094] In one embodiment, the composition is formulated into granules, a water-soluble powder, or a wettable powder.

[0095] Another embodiment of the present disclosure provides a granular biofertilizer composition comprising an endophytic bacterium selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, and one or more mycorrhizae. In one embodiment, the mycorrhizal vegetative propagule comprises an endomycorrhizal vegetative propagule. In another embodiment, the mycorrhizal vegetative propagule comprises an ectomycorrhizal vegetative propagule. In one embodiment, the mycorrhizal vegetative propagule comprises a combination of an endomycorrhizal vegetative propagule and an ectomycorrhizal vegetative propagule. In one embodiment, the composition further comprises a plant extract, a stabilizer, and an agriculturally acceptable excipient. In one embodiment, the plant extract is an extract from a plant belonging to the genus Dodonaea. In one embodiment, the stabilizer serves as a carbon source that helps improve the viability of endophytic bacteria and mycorrhizal vegetative growths and improve the shelf life of the composition. In some embodiments, the plant extract of Dodonaea viscosa also exerts a stabilizing effect on endophytic bacteria and mycorrhizal vegetative growths, thereby helping to improve the shelf life of the composition.

[0096] In one embodiment, the composition comprises (a) an endophytic bacterium selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, in an amount ranging from about 0.1% to about 30% by weight; (b) a mycorrhizal vegetative growth in an amount ranging from about 5% to about 40% by weight of the composition; (c) an extract of Dodonaea viscosa in an amount ranging from about 0.2% to about 20% by weight of the composition; (d) a stabilizer in an amount ranging from about 0.2% to about 80% by weight of the composition; and (e) an agriculturally acceptable excipient. In one embodiment, the stabilizer is selected from the group including humic acid, cellulose, dextrose, lactose, methylcellulose, xanthan gum, polyvinylpyrrolidone (PVP), Tween (polysorbate), gum arabic, or combinations thereof. In one embodiment, the stabilizer comprises humic acid and methylcellulose. In another embodiment, the stabilizer comprises humic acid and cellulose.

[0097] In one embodiment, the composition comprises (a) an amount ranging from about 0.1% to about 30% by weight of endophytic bacteria selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof; (b) an amount ranging from about 5% to about 40% by weight of the composition of mycorrhizal vegetative growths; and (c) an amount ranging from about 0.2% to about 20% by weight of Dodonaea The composition comprises (d) an extract of viscosa, (e) humic acid in an amount ranging from about 1% to about 50% by weight of the composition, (f) cellulose or methylcellulose in an amount ranging from about 0.1% to about 5% by weight of the composition, and (g) an agriculturally acceptable excipient in an amount sufficient to prepare the composition described above.

[0098] In one embodiment, the disclosure provides a wettable biofertilizer composition comprising an endophytic bacterium selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, and one or more mycorrhizae. In one embodiment, the mycorrhizal vegetative propagule comprises an endomycorrhizal vegetative propagule. In another embodiment, the mycorrhizal vegetative propagule comprises an ectomycorrhizal vegetative propagule. In one embodiment, the mycorrhizal vegetative propagule comprises a combination of an endomycorrhizal vegetative propagule and an ectomycorrhizal vegetative propagule. In one embodiment, the composition further comprises a stabilizer and an agriculturally acceptable excipient, the stabilizer being selected from humic acid, cellulose, methylcellulose, xanthan gum, polyvinylpyrrolidone (PVP), Tween, and gum arabic, or a combination thereof.

[0099] In one embodiment, the composition comprises (a) an endophytic bacterium selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, in an amount ranging from about 0.1% to about 30% by weight; (b) a mycorrhizal vegetative growth in an amount ranging from about 5% to about 40% by weight of the composition; (c) a stabilizer in an amount ranging from about 0.2% to about 80% by weight of the composition; and (d) an agriculturally acceptable excipient. In one embodiment, the stabilizer is selected from the group including humic acid, cellulose, methylcellulose, xanthan gum, polyvinylpyrrolidone (PVP), Tween (polysorbate), gum arabic, or a combination thereof. In one embodiment, the stabilizer comprises humic acid and methylcellulose. In another embodiment, the stabilizer comprises humic acid and cellulose.

[0100] In one embodiment, the composition comprises (a) an endophytic bacterium selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, in an amount ranging from about 0.1% to about 30% by weight; (b) mycorrhizal vegetative growths in an amount ranging from about 5% to about 40% by weight of the composition; (c) humic acid in an amount ranging from about 1% to about 50% by weight of the composition; (d) cellulose or methylcellulose in an amount ranging from about 0.1% to about 5% by weight of the composition; and (e) an agriculturally acceptable excipient in an amount sufficient to prepare the above composition.

[0101] In some embodiments, the amount of humic acid in the composition is in the range of about 5% to about 50% by weight, about 5% to about 45% by weight, about 5% to about 40% by weight, about 5% to about 35% by weight, about 5% to about 30% by weight, about 5% to about 25% by weight, about 5% to about 20% by weight, about 10% to about 50% by weight, about 10% to about 40% by weight, about 10% to about 30% by weight, about 20% to about 50% by weight, about 20% to about 40% by weight, or about 25% to about 50% by weight.

[0102] In some embodiments, the amount of cellulose or methylcellulose in the composition is in the range of about 0.1% to about 5% by weight, about 0.1% to about 4.5% by weight, about 0.1% to about 4% by weight, about 0.1% to about 3.5% by weight, about 0.1% to about 3% by weight, about 0.1% to about 2.5% by weight, about 0.1% to about 2% by weight, about 0.1% to about 1.5% by weight, and about 0.1% to about 1% by weight.

[0103] One embodiment of the present disclosure provides a method for preparing a biofertilizer composition comprising endophytic bacteria selected from Pseudomonas fluorescens, Bacillus subtilis, Bacillus amyloliquefaciens, Pseudomonas putida, Paenibacillus polymyxa, or a mixture thereof, and one or more mycorrhizae.

[0104] One embodiment of the present disclosure provides a method for preparing a biofertilizer composition comprising an endophytic bacterium selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, and one or more mycorrhizae.

[0105] One embodiment provides a method for improving the health, yield and vitality of a plant, the method comprising contacting a plant or a part thereof, plant propagation material, a place or medium in which a plant grows, or a growing area with a combination of biofertilizers or a composition containing the above combination, wherein the combination comprises endophytic bacteria selected from Pseudomonas fluorescens, Bacillus subtilis, Bacillus amyloliquefaciens, Pseudomonas putida, Paenibacillus polymyxa, or a mixture thereof, and one or more mycorrhizae.

[0106] One embodiment provides a method for improving the health, yield, and vitality of a plant, the method comprising contacting a plant or a part thereof, plant propagation material, a place or medium in which the plant grows, or a growing area with a combination of biofertilizers, or a composition containing the above combination, wherein the combination comprises endophytic bacteria selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, and one or more mycorrhizae.

[0107] Agricultural benefits may be imparted by treating seeds, seedlings, plants, plant parts, soil, or combinations thereof with the compositions or formulations of the Disclosure under conditions that result in association between the compositions and seeds, plants, plant parts, soil, or combinations thereof. Application of the formulations to a target is achieved using any delivery method known in the art, including, but not limited to, spraying, fumigation, granular application, injection, misting, seed treatment, spraying, dipping, or coating.

[0108] In one embodiment, the combination or composition of the present disclosure may be applied to seeds or any plant propagation material for seed treatment.

[0109] In one embodiment, the present disclosure provides a method for treating plant seeds, comprising applying a combination of biofertilizers, or a composition comprising the above-mentioned combination, wherein the combination comprises an endophytic bacterium selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, and one or more mycorrhizae. In one embodiment, the method comprises applying a combination of biofertilizers, or a composition containing the above-mentioned combination, the combination comprising an endophytic bacterium selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, and one or more mycorrhizae.

[0110] The term "plant propagation material" is understood to refer to all reproductive parts of a plant, such as seeds, as well as growing plant material such as cuttings and tubers (e.g., potatoes), which can be used for plant propagation. This includes seeds, roots, fruits, tubers, bulbs, rhizomes, shoots, sprouts, and other parts of the plant. It also includes seedlings and young plants that should be transplanted after germination or emergence from the soil.

[0111] In one embodiment, the combination or composition of the Disclosure may be applied to crops or seeds. Examples of crops to which the combinations and compositions of the Disclosure may be used include, but are not limited to, corn, rice, wheat, barley, rye, oats, sorghum, cotton, soybeans, peanuts, buckwheat, beets, rapeseed, sunflowers, sugarcane, tobacco, etc.; nightshade vegetables such as eggplant, tomato, pimento, pepper, and potato; cucurbit vegetables such as cucumber, pumpkin, zucchini, watermelon, melon, and squash; radish, turnip, horseradish, kohlrabi Brassicaceae vegetables such as cabbage, Chinese cabbage, mustard greens, broccoli, and cauliflower; Asteraceae vegetables such as burdock, garland chrysanthemum, artichoke, and lettuce; Liliaceae vegetables such as leeks, onions, garlic, and asparagus; Apiaceae vegetables such as carrots, parsley, celery, and parsnips; Chenopodiaceae vegetables such as spinach and Swiss chard; Lamiaceae vegetables such as perilla, mint, and basil; Strawberries, sweet potatoes, and yams (Dioscorea) Taro (Japonica), etc.; Flowers, ornamental plants, grass; Pomelo such as apples, pears, and quince; Stone fruits such as peaches, plums, nectarines, Japanese apricots, cherries, apricots, and prunes; Citrus fruits such as oranges, lemons, limes, and grapefruits; Nuts such as chestnuts, walnuts, hazelnuts, almonds, pistachios, cashews, and macadamia nuts; Berries such as blueberries, cranberries, blackberries, and raspberries; Grapes, persimmons, olives, plums, bananas, coffee, dates, coconuts, etc.; Trees other than fruit trees; Tea, mulberry, flowering plants, ash, birch, dogwood, eucalyptus, ginkgo, lilac, maple, oak, poplar, judas tree, sweetgum (Liquidambar formosana), plane tree, zelkova, Japanese cedar (Japanese Examples include trees such as fir, poison hemlock, juniper, pine, spruce, and Japanese yew (Taxus cuspidata), belonging to the Arborvitae family.

[0112] In one embodiment, crops to which the combinations and compositions of the present disclosure may be used include rice, wheat, barley, maize, millet, fiber crops, vegetable crops, fruit crops, spices, flowers, fodder, and plants.

[0113] The combinations and compositions disclosed herein may be used at the time of sowing or transplanting.

[0114] The combinations and compositions disclosed herein can be applied to growing areas by conventional methods such as ground sprayers, granule spreaders, sprinkling (drenching), drip irrigation, spraying, atomization, broadcasting, powdering, foaming, coating, aerial spraying, aerial application, and, without limitation, by methods utilizing modern technologies such as drones and robots, and by other conventional means known to those skilled in the art.

[0115] In one embodiment, the combination or composition of the present disclosure may be mixed with or applied together with other pesticide active ingredients. The other pesticide active ingredients may be selected from fertilizers, micronutrients, mite control agents, algaecides, feeding inhibitors, bird killers, fungicides, bird repellents, sterilizers, fungicides, herbicides, insect attractants, insect repellents, insecticides, mammal repellents, mating disruptors, mollusk control agents, nematicides, plant activators, plant growth regulators, rodenticides, synergists, antivirals, derivatives thereof, biological regulators, or mixtures thereof. The above mixtures may be pre-mixed compositions or may be tank-mixed at the time of application. The combination or composition of the present disclosure may be applied together with other pesticide active ingredients or separately and sequentially.

[0116] In preferred embodiments, other pesticide active ingredients are selected from the group consisting of acephate + cypermethrin, copper sulfate + mancozeb, propineb, thiamethoxam, forcysteine, Foltron®, Wuxal®, imidacloprid, carbendazim + mancozeb, spinetoram, and orthosilicic acid (OSA).

[0117] Another embodiment of the present disclosure provides the use of a biofertilizer composition comprising endophytic bacteria selected from Pseudomonas fluorescens, Bacillus subtilis, Bacillus amyloliquefaciens, Pseudomonas putida, Paenibacillus polymyxa, or a mixture thereof, and one or more mycorrhizae, for improving plant health, yield, and vitality.

[0118] Another embodiment of the present disclosure provides the use of a combination of biofertilizers for improving plant health, yield, and vitality, comprising endophytic bacteria selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, and one or more mycorrhizae.

[0119] One embodiment of the present disclosure also provides a kit comprising a component comprising endophytic bacteria selected from Pseudomonas fluorescens, Bacillus subtilis, Bacillus amyloliquefaciens, Pseudomonas putida, Paenibacillus polymyxa, or a mixture thereof, and another component comprising one or more mycorrhizae for improving plant health, yield, and vitality.

[0120] One embodiment of the present disclosure also provides a kit comprising a component comprising an endophytic bacterium selected from Pseudomonas fluorescens deposited as MTCC 25519, Bacillus subtilis deposited as MTCC 25520, Bacillus amyloliquefaciens deposited as MTCC 25518, Pseudomonas putida deposited as MTCC 25607, Paenibacillus polymyxa deposited as MTCC 25609, or a mixture thereof, and another component comprising one or more mycorrhizae for improving plant health, yield, and vitality.

[0121] In one embodiment, the kit may be divided into multiple components of endophytic bacteria and one or more mycorrhizae, which are mixed together, sequentially, or simultaneously. [Examples]

[0122] Example 1: Endophytic bacteria + mycorrhizal WP composition, and their effectiveness in relation to root length, shoot length, and colony formation by mycorrhizal fungi.

[0123] Nine WP compositions (Compositions 1-9) containing endophytic bacteria and endomycorrhizal species Rhizophagus irregularis, Rhizophagus clarus, Gigaspora margarita, and Dentiscutata heterogama, and ectomycorrhizal species Pisolithus tinctorius and Piriformospora indica were prepared for application to tomatoes and pigeon peas. A control WP composition (Composition 10) containing only the endomycorrhizal species Rhizophagus irregularis, Rhizophagus clarus, Gigaspora margarita, and Dentiscutata heterogama and the ectomycorrhizal species Pisolithus tinctorius and Piriformospora indica was prepared for comparison. Compositions 1-10 are provided in Table 1. Application rate of composition: 100g / acre Application time: At the time of transplanting Number of applications: Once at the time of transplantation

[0124] The effectiveness of the composition on root length, shoot length, and mycorrhizal fungal colony formation was evaluated 45 days after application.

[0125] [Table 1]

[0126] [Table 2]

[0127] The above tests show that when endophytic bacteria are combined with mycorrhizae in the composition, there is a significant improvement in colony formation by mycorrhizal fungi, as well as in shoot length and root length, in pigeon peas and tomatoes.

[0128] Example 2: Endophytic bacteria + mycorrhizal granule composition, and their effectiveness in relation to root length, shoot length, and colony formation by mycorrhizal fungi.

[0129] Nine granular compositions (compositions 11-19) containing endomycorrhizal bacteria and the endomycorrhizal species Rhizophagus irregularis, Rhizophagus clarus, Gigaspora margarita, and Dentiscutata heterogama, as well as the ectomycorrhizal species Pisolithus tinctorius and Piriformospora indica, were prepared for application to tomatoes, peppers, and eggplants. A control granular composition (composition 20) containing only the endomycorrhizal species Rhizophagus irregularis, Rhizophagus clarus, Gigaspora margarita, and Dentiscutata heterogama, and the ectomycorrhizal species Pisolithus tinctorius and Piriformospora indica, was prepared for comparison. Compositions 11-20 are provided in Table 3. Application rate of composition: 100g / acre Application time: At the time of transplanting Number of applications: Once at the time of transplantation

[0130] The effectiveness of the composition on root length, shoot length, and mycorrhizal fungal colony formation was evaluated 45 days after application.

[0131] [Table 3]

[0132] [Table 4]

[0133] The above tests show that when endophytic bacteria are combined with mycorrhizae in the composition, there is a significant improvement in mycorrhizal colony formation, shoot length, and root length in chili peppers, eggplants, and tomatoes.

Claims

1. A combination of biofertilizers comprising endophytic bacteria selected from Pseudomonas fluorescens, Bacillus subtilis, Bacillus amyloliquefaciens, Pseudomonas putida, Paenibacillus polymyxa, or mixtures thereof, and one or more mycorrhizae.

2. The biofertilizer combination according to claim 1, wherein Pseudomonas fluorescens is deposited as MTCC 25519, Bacillus subtilis is deposited as MTCC 25520, Bacillus amyloliquefaciens is deposited as MTCC 25518, Pseudomonas putida is deposited as MTCC 25607, and Paenibacillus polymyxa is deposited as MTCC 25609.

3. The biofertilizer combination according to claim 1, wherein the combination includes at least two types of endophytic bacteria.

4. The biofertilizer combination according to claim 1, wherein the mycorrhizae are endomycorrhizae, ectomycorrhizae, or a combination thereof.

5. The biofertilizer combination according to claim 4, wherein the mycorrhizae include a mycorrhizal vegetative propagator selected from an endomycorrhizal vegetative propagator, an ectomycorrhizal vegetative propagator, or a combination thereof.

6. The biofertilizer combination according to claim 5, wherein the mycorrhizal vegetative growth includes an endomycorrhizal vegetative growth selected from the group consisting of the genus Glomus, Entrophospora, Funneliformis, Rhizophagus, Septoglobus, Acaulospora, Archaeospora, Paraglobus, Gigaspora, Pacispora, Sclerocystis, Dentiscutata, Scutellospora, and combinations thereof.

7. The biofertilizer combination according to claim 6, wherein the mycorrhizal vegetative growth includes an endomycorrhizal vegetative growth selected from the group consisting of Rhizophagus irregularis, Rhizophagus clarus, Gigaspora margarita, Scutellospora heterogama, Glomus mossae, Septoglomus viscosum, Funneliformis mossae, and combinations thereof.

8. The biofertilizer combination according to claim 5, wherein the mycorrhizal vegetative growth comprises ectomycorrhizal vegetative growth selected from Laccaria, Pisolithus, Piriformospora, Scleroderma, Rhizopogon, and combinations thereof.

9. The biofertilizer combination according to claim 8, wherein the mycorrhizal vegetative growth comprises ectomycorrhizal vegetative growth selected from Piriformospora indica, Laccaria bicolor, Laccaria lacata, Rhizopogon roseolus, Laccaria proxima, Pisolithus arhizus, Pisolithus tinctorius, and combinations thereof.

10. The biofertilizer combination according to claim 1, wherein the endophytic bacteria and mycorrhizae are mixed in a ratio of 1:100 to 100:

1.

11. A biofertilizer composition comprising the combination described in claim 1.

12. The biofertilizer composition according to claim 11, wherein the endophytic bacteria are present in an amount ranging from about 0.1% to about 50% by weight of the total composition.

13. The biofertilizer composition according to claim 11, wherein the composition comprises endomycorrhizae, ectomycorrhizae, or a combination thereof in an amount ranging from about 0.1% by weight to about 90% by weight of the composition.

14. The biofertilizer composition according to claim 11, wherein the composition further comprises one or more agriculturally acceptable excipients.

15. The biofertilizer composition according to claim 11, wherein the composition further comprises a plant extract.

16. The biofertilizer composition according to claim 11, wherein the composition is formulated into granules, a water-soluble powder, or a wettable powder.

17. A method for improving the health, yield and vitality of a plant, wherein the method comprises bringing a plant or a part thereof, plant propagation material, a place or medium in which a plant grows, or a growing area, into contact with a combination of biofertilizers described in claim 1, or a biofertilizer composition containing such combination.

18. The biofertilizer combination according to claim 1, wherein the combination, or a composition comprising the combination, is mixed with or applied together with other pesticide active ingredients selected from fertilizers, micronutrients, mite control agents, algaecides, feeding inhibitors, bird killers, fungicides, bird repellents, sterilizers, fungicides, herbicide safeners, herbicides, insect attractants, insect repellents, insecticides, mammal repellents, mating disruptors, mollusk control agents, nematicides, plant activators, plant growth regulators, rodenticides, synergists, antiviral agents, derivatives thereof, biological regulators, or mixtures thereof.

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