# US 2021/0276927 A1 · Novel paenibacillus polymyxa and uses thereof

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- Number: US 2021/0276927 A1
- Document ID: US20210276927A1
- Status: Published application
- Kind: A1
- Application number: 16/967,990
- Filed: 2018-02-07
- Published: 2021-09-09
- Earliest claimed: 2018-02-07
- Assignees: Valagro S.P.A.
- Applicants: Valagro S.P.A.
- Inventors: Matteo DI MUZIO; Ilaria LEBANO; Juan Fernando MEJIA DE LOS RIOS; Donata DI TOMMASO; Alberto PIAGGESI; Prem WARRIOR
- Web page: https://patentgrep.com/patent/US20210276927A1

## CPC

- C05F 11/08: Organic fertilisers containing added bacterial cultures, mycelia or the like
- C05B 19/00: Granulation or pelletisation of phosphatic fertilisers, other than slag (granulating apparatus B01J2/00; granulating slag C04B)
- C05F 1/007: {from derived products of animal origin or their wastes, e.g. leather, dairy products}
- C05F 5/002: {Solid waste from mechanical processing of material, e.g. seed coats, olive pits, almond shells, fruit residue, rice hulls}
- C05F 11/02: from peat, brown coal, and similar vegetable deposits
- C12N 1/205: Bacterial isolates
- C12R 2001/12: Bacillus polymyxa {; Paenibacillus polymyxa}
- C05F: ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05B: PHOSPHATIC FERTILISERS
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS

## Abstract

The present invention refers to new bacterial strains, a consortium comprising said strains, a composition comprising at least one of these novel bacterial strains, preferably in combination with a plant biostimulant and their use in agriculture, preferably, to improve nutrients up-take in plants and therefore to ameliorate plant growth and/or development.

## Claims

### Claim at position 1

1-35. (canceled)

### Claim 36

A microbial consortium comprising
an isolated bacterial strain belonging to the genus Bacillus species simplex characterized by at least one species-specific sequence selected from: SEQ ID NO: 1-4 or sequences having at least 80-99% identity, wherein SEQ ID NO: 1 refers to 16S rRNA gene, SEQ ID NO: 2 refers to gapA gene, SEQ ID NO: 3 refers to uvrA gene and SEQ ID NO: 4 refers to pgk gene, and
an isolated bacterial strain belonging to the genus Paenibacillus, species polymyxa by at least one species-specific sequence selected from: SEQ ID NO: 5-7 or sequences having at least 80-99% identity, wherein SEQ ID NO: 5 refers to 16S rRNA gene, SEQ ID NO: 6 refers to rpoB gene, and SEQ ID NO: 7 refers to nifH gene.

### Claim 37 (depends on 36)

The microbial consortium of claim 36, wherein
the isolated bacterial strain belonging to the genus Bacillus species simplex has been deposited at the DSMZ with the accession number DSM32459 and the following name: Bacillus simplex strain VMC10/70, and
the isolated bacterial strain belonging to the genus Paenibacillus, species polymyxa has been deposited at the DSMZ with the accession number DSM32460 and the following name: Paenibacillus polymyxa strain VMC10/96.

### Claim 38 (depends on 36)

The microbial consortium of claim 36, wherein the isolated bacterial strains are used in a form selected from the group consisting of: fresh bacteria, frozen bacteria, dry bacteria, lyophilized bacteria, liquid suspension of bacteria, encapsulated bacteria in the form of spores, living bacteria, culture medium, extract of bacteria, supernatant, lysate of bacteria, fraction of bacteria, metabolites derived from said bacteria, and any combination thereof.

### Claim 39 (depends on 36)

The microbial consortium according to claim 36, wherein the isolated bacterial strains are mutated and/or edited.

### Claim 40 (depends on 36)

The microbial consortium according to claim 36, further comprising microorganisms.

### Claim 41 (depends on 40)

The microbial consortium according to claim 40, wherein the microorganisms are bacteria selected the from the group consisting of PGPR or rhizobacteria, yeasts, mycorrhizae, fungi, and derivatives and combinations thereof, wherein:
the PGPR is selected from the group consisting of Aeromonas rivuli, Agromyces fucosus, Bacillus spp. Bacillus mycoides, Bacillus licheniformis, Bacillus subtilis, Bacillus megaterium, Bacillus pumilus, Bacillus safensis, Microbacterium sp., Nocardia globerula, Stenotrophomonas spp., Pseudomonas spp, Pseudomonas fluorescens, Pseudomonas fulva, Pseudoxanthomonas dajeonensis, Rhodococcus coprophilus, Sphingopyxis macrogoltabida, Streptomyces spp., Enterobacter spp., Azotobacter spp., Azospiriullum spp., Rhizobium spp., Herbaspirillum spp., Lactobaccillus spp., Lactobacillus acidophilus, Lactobacillus buchneri, Lactobacillus delbrueckii, Lactobacillus johnsonii, Lactobacillus murinus, Lactobacillus paraplantarum, Lactobacillus pentosus, Lactobacillus plantarum, Lactococcus tactis, and combinations thereof;
the yeast is selected from the group consisting of Candida spp., Candida tropicalis, Saccharomyces spp., Saccharomyces bayanus, Saccharomyces boulardii, Saccharomyces cerevisiae, Saccharomyces exiguous, Saccharomyces pastorianus, Saccharomyces pombe, and combinations thereof;
the mycorrhiza is selected from the group consisting of Glomus spp., Rhizophagus spp., Septoglomus spp., Funneliformis spp., and combinations thereof; and
the fungus is selected from the group consisting of Trichoderma spp., Trichoderma atroviride, Trichoderma viride, Trichoderma afroharzianum, Paecilomyces spp., Beauveria bassiana, Metarhizium spp., Lecanicillium lecanii, Penicillium spp., Aspergillus spp., Conythyrium minitans, Pythium spp, and combinations thereof.

### Claim 42 (depends on 36)

The microbial consortium according to claim 36, wherein the strain is used as a fresh or frozen sample or dry, lyophilized or in liquid suspension, or encapsulated in the form of living and/or dead and/or killed cells and/or as spores.

### Claim 43 (depends on 40)

The microbial consortium according to any one of claim 40, wherein the ratio between the strain and the further microorganisms ranges from 1:10 to 1:100.

### Claim 44 (depends on 36)

The microbial consortium according to claim 36, further comprising a biostimulant.

### Claim 45 (depends on 44)

The microbial consortium according to claim 44, wherein the biostimulant is selected from the group consisting of an extract of algae, an extract of microalgae, an extract of plant, a humic acid, a fulvic acid, animal byproducts, or combinations thereof.

### Claim 46 (depends on 45)

The microbial consortium according to claim 45, wherein:
the algae are a brown algae selected from the group consisting of Ascophyllum nodosum, Ecklonia maxima, Laminaria saccharina, Laminaria digitata, Fucus spiralis, Fucus serratus, F. vesiculosus, Macrocystis spp., Pelvetia canaliculata, Himantalia elongata, Undaria pinnatifida, Sargassum spp, and combinations thereof;
the microalgae are selected from the group consisting of Spirulina, Scenedesmus, Nannochloropsis, Haematococcus, Chlorella, Phaeodactylum, Arthrospyra, Tetraselmis, Isochrysis, Synechocystis, Clamydomonas, Parietochloris, Desmodesmus, Neochloris, Dunaliella, Thalassiosira, Pavlova, Navicula, Chaetocerous, and combinations thereof; and
the plant is selected from the group consisting of beet, sugar cane, alfalfa, maize, brassica, halophytes, soya, wheat, yucca, quillaja, hop, coffee, citrus, olive, and combinations thereof.

### Claim 47 (depends on 36)

A medium, an extract, a supernatant, a lysate, a fraction, or a metabolite obtained/obtainable by culturing the microbial consortium of claim 36, wherein said medium, extract, supernatant, lysate, fraction, or metabolite comprises the cultured bacteria or is a bacteria-free medium.

### Claim 48 (depends on 36)

An agricultural composition, comprising the microbial consortium of claim 36, and an agriculturally compatible carrier.

### Claim 49 (depends on 47)

An agricultural composition, comprising the medium, extract, supernatant, lysate, fraction or metabolite of claim 47, and an agriculturally compatible carrier.

### Claim 50 (depends on 36)

The microbial consortium according to claim 36, formulated as water-soluble concentrates, dispersable concentrates, emulsifiable concentrates, emulsions, suspensions, microemulsion, gel, microcapsules, granules, ultralow volume liquid, wetting powder, dustable powder, or seed coating formulations.

### Claim 51 (depends on 47)

The medium, extract, supernatant, lysate, fraction or metabolite according to claim 47, formulated as water-soluble concentrates, dispersable concentrates, emulsifiable concentrates, emulsions, suspensions, microemulsion, gel, microcapsules, granules, ultralow volume liquid, wetting powder, dustable powder, or seed coating formulations.

### Claim 52 (depends on 49)

The agricultural composition according to claim 49, formulated as water-soluble concentrates, dispersable concentrates, emulsifiable concentrates, emulsions, suspensions, microemulsion, gel, microcapsules, granules, ultralow volume liquid, wetting powder, dustable powder, or seed coating formulations.

### Claim 53 (depends on 42)

The microbial consortium according to claim 42, wherein the strain is the Bacillus simplex strain VMC10/70 and Paenibacillus polymyxa strain VMC10/96.

### Claim 54 (depends on 42)

The microbial consortium according to claim 42, wherein the ratio between the strain and further microorganisms is approximately 1:1.

### Claim 55 (depends on 47)

The medium, an extract, a supernatant, a lysate, a fraction, or a metabolite of claim 47, wherein the extract is a bacteria-free extract.