EP4168532A1 - Biostimulans auf bakterienbasis zur besseren anpassung von pflanzen an hydrische und osmotische belastungen - Google Patents

Biostimulans auf bakterienbasis zur besseren anpassung von pflanzen an hydrische und osmotische belastungen

Info

Publication number
EP4168532A1
EP4168532A1 EP21740260.1A EP21740260A EP4168532A1 EP 4168532 A1 EP4168532 A1 EP 4168532A1 EP 21740260 A EP21740260 A EP 21740260A EP 4168532 A1 EP4168532 A1 EP 4168532A1
Authority
EP
European Patent Office
Prior art keywords
water
bacteria
plant
strains
soils
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
EP21740260.1A
Other languages
English (en)
French (fr)
Inventor
Mohamed Hafidi
Lamfeddal Kouisni
Ahmed NAFIS
Yedir Ouhdouch
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
OCP SA
Original Assignee
OCP SA
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by OCP SA filed Critical OCP SA
Publication of EP4168532A1 publication Critical patent/EP4168532A1/de
Pending legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P21/00—Plant growth regulators
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/465—Streptomyces
    • C12R2001/47—Streptomyces albus

Definitions

  • Bacteria-based biostimulant for better adaptation of plants to water and osmotic stress Bacteria-based biostimulant for better adaptation of plants to water and osmotic stress.
  • Figure 1 Diagram of the technique for evaluating the degree of hydrophobicity of a sandy soil (a) and determination of the volume of water retained (b)
  • FIG. 2 Elemental analysis of sand at low altitude a: BNT: untreated sand, b: BT: sand treated with butanone. (% CK in BNT is 29 against 14 in BT)
  • Figure 3a Macroscopic and microscopic aspects of isolates (NI, N3 & N4) degrading commercial waxes.
  • Figure 3b Phylogenetic tree of bacterial strains capable of degrading waxes (NI, N4 and N3) retained for this invention.
  • Figure 4 Examples of use of insoluble forms of K and P expressed by the extent of growth of strains (NI, N4 and N3) on screening media (Aleksandrov & NBRIP)
  • FIG. 5 Protocol followed for the evaluation of root colonization of alfalfa and Aradopsis seedlings by strains (NI, N4 and N 3).
  • Graph 1 PGP traits of NI, N4 and N4 strains: Germination index, TCP solubilization of Mica and auxin production
  • Potassium (K) is an essential element involved in water management at the plant level. Indeed, For plants, potassium (K) plays an important role in water and osmotic status: (i) by regulating the opening and closing of stomata, (ii) by the osmotic adjustment of the plant (turgor) , (iii) and by root growth. In addition to these 3 functions, the University of Halle has demonstrated a significant impact of potassium on the useful water reserve of the soil (Damm 2012).
  • This invention describes obtaining a plant biostimulant based on bacteria isolated from the root system of desert plants (rhizospheric soils).
  • the choice of this habitat is not trivial.
  • Our goal was to understand how the plants in this stressed habitat manage to resist and cope with the lack of water.
  • These bacteria which help desert plants are able to reduce water and osmotic stress thanks to their following essential properties:
  • BDC waxes
  • BDC + BSK bacterial inoculum based on the optimization of certain biological properties in these bacteria
  • BDC + BSK- Plant the role of the partnership (BDC + BSK- Plant) on the development of the host plant will be optimized by intimately combining the impact of these (BCD + BSK) on the plant directly (water, mineral, hormonal effect, etc. ) and indirectly (alteration of potassium and inorganic phosphate).
  • BCD + BSK Bacillus subtilis .
  • These bacteria are cultivated on liquid nutrient medium obtained from soybean flour juice after filtration (3g / L of water). After a period of culture depending on the physiological characteristics of (BCD + BSK), the propagules are recovered by filtration, washed with sterile water to remove all chemical traces from the nutrient medium used then ground in a Waring Blender and stored at room temperature and at 4 ° C.
  • Granules of potassium (Mica) (50 ⁇ m diameter) or commercial perlite are immersed in a sterile solution composed (i) of soybean flour juice (3 g / L), previously sterilized (ii) of propagules (1 g dry weight per liter of mixture).
  • mice and / or perlite granules impregnated with the bacteria are incubated at 28 ° C. for 2 days.
  • the perlite and Mica granules thus colonized by (BCD + BSK) forming propagules on the surface are then dried in air and placed in flasks to store them at room temperature and / or at 4 ° C.
  • Another advantage of this invention is the fact that the seeds or seeds can be inoculated directly with the propagules of (BCD + BSK) by the method of seed bacterization.
  • a new method of isolating bacteria, a new biotechnology and a new biostimulant isolating bacteria, a new biotechnology and a new biostimulant.
  • BDC Effect Optimization
  • Rhizospheric soils collected near the roots of desert plants have very low water repellency (with a time of less than 10 seconds).
  • bare sands far from the root system of plants show pronounced hydrophobicity with a time that is between 250-260 seconds.
  • Method 1 The rhizospheric or non-rhizospheric sand grains are washed with sterile distilled water and dried were seeded (deposited) directly in a minimum culture medium (MM) without any source of organic carbon.
  • the composition of MM consists of the following ingredients (for 1 liter): (KNO 3 2g, K 2 HPO 4 lg, MgS0 4 0.5 g, NaCl 0.5g, CaC0 3g, FeS0 4 0.01g and noble agar or agarose 14g
  • the only source of organic carbon is that which surrounds the grains of sand.
  • Method 2 From 200g of rhizospheric or non-rhizospheric rice, dried, treated with butanone to extract waxes and propagules from microorganisms. After evaporation of the butanone, the suspension of propagules was seeded on the MM + 5 g of commercial wax (for 1 liter MM + agar-agar not usable by bacteria). Three waxes were used.
  • the colonies developed were purified and tested for their ability to degrade a wide range of commercial waxes, namely, Carnauba wax, Candelilla wax and Mimosa wax.
  • the controls were used: MM + Agar-agar and wax + agar-agar
  • N3 isolate is identified as Voyl anyium bmchyspomm
  • N 4 isolate identified as Streptomyces mutabilis
  • NI is identified as Streptomyces acumycini.
  • PGP plant growth promotion
  • Aleksandrov and NBRIP media without any soluble source of K and P represent negative controls.
  • Aleksandrov and NBRIP media with soluble sources of K and P represent positive controls.
  • TCP source of P
  • Mica used for the solubilization of K and P in the same culture medium.
  • An example of the growth obtained is given in figure 4.
  • the solubilization performance of the insoluble forms of K and P of these isolates was measured via the analysis of the culture juices by ISP (Inductively Coupled Plasma).
  • ISP Inductively Coupled Plasma
  • the AIA assay was performed in the juice of cultures as well. Germination tests were carried out on Arabidopsis taliana seeds as a model plant. Before germinating the seeds, the latter were disinfected and bacterized by stem cells (NI, N4 and N 3) cultured for three days in Bennett's medium in order to produce sufficient biomass. Subsequently, the seeds of the model plant were cultivated under optimal growth conditions and the results are noted after 5 days.
  • the sandy soil used is characterized by the physicochemical parameters recorded in graph n ° 2.
  • the strains (NI, N4 and N3) are cultivated on liquid nutrient medium obtained from soybean flour juice after filtration (3 g / L of water). After 3 days, the propagules are recovered by filtration, washed with sterile water to remove all chemical traces from the nutrient medium used, then ground in a Waring Blender then dried. 100 g of the water-repellent sand of composition indicated in Table 2 are inoculated with 10 6 CFU (colony-forming unit) of each strain. The dried propagules were mixed well with the sand.
  • the N3 strain identified as Poyl angium bmchyspomm showed a biostimulating capacity of alfalfa seedlings with a yield of more than 50% compared to the commercial expansive vermiculite control product (Graph 5).
  • the results of alfalfa plants were found to be more interesting in terms of the biomass yield of the obtained plant material.
  • the biostimulatory effect of the N3 strain is the most important. In fact, this strain has made it possible to increase the yield in terms of plant biomass by more than 50% and it is the same strain that has made it possible to obtain the increase in water retention of about 10% of the soil. used.
  • the potassium contents of the different parts of the alfalfa seedlings were improved for the seedlings inoculated with the N3 strain.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Plant Pathology (AREA)
  • Virology (AREA)
  • Environmental Sciences (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Pest Control & Pesticides (AREA)
  • Microbiology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Biochemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Botany (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biomedical Technology (AREA)
  • Agronomy & Crop Science (AREA)
  • Dentistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Chemical & Material Sciences (AREA)
  • Soil Conditioners And Soil-Stabilizing Materials (AREA)
  • Cultivation Of Plants (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
EP21740260.1A 2020-06-22 2021-06-18 Biostimulans auf bakterienbasis zur besseren anpassung von pflanzen an hydrische und osmotische belastungen Pending EP4168532A1 (de)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
MA50114A MA50114B1 (fr) 2020-06-22 2020-06-22 Biostimulant à base de bactéries pour une meilleure adaptation des ?Plantes aux stress hydrique et osmotique?
PCT/MA2021/000012 WO2021261984A1 (fr) 2020-06-22 2021-06-18 Biostimulant à base de bactéries pour une meilleure adaptation des plantes aux stress hydrique et osmotique

Publications (1)

Publication Number Publication Date
EP4168532A1 true EP4168532A1 (de) 2023-04-26

Family

ID=76859690

Family Applications (1)

Application Number Title Priority Date Filing Date
EP21740260.1A Pending EP4168532A1 (de) 2020-06-22 2021-06-18 Biostimulans auf bakterienbasis zur besseren anpassung von pflanzen an hydrische und osmotische belastungen

Country Status (8)

Country Link
EP (1) EP4168532A1 (de)
KR (1) KR20240010707A (de)
CN (1) CN116133525A (de)
AU (1) AU2021296688A1 (de)
CA (1) CA3189825A1 (de)
MA (1) MA50114B1 (de)
WO (1) WO2021261984A1 (de)
ZA (1) ZA202300601B (de)

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2988764A1 (en) * 2015-06-08 2016-12-15 Indigo Agriculture, Inc. Streptomyces endophyte compositions and methods for improved agronomic traits in plants

Also Published As

Publication number Publication date
AU2021296688A1 (en) 2023-02-23
WO2021261984A1 (fr) 2021-12-30
CN116133525A (zh) 2023-05-16
CA3189825A1 (en) 2021-12-30
MA50114B1 (fr) 2022-04-29
MA50114A1 (fr) 2021-12-31
KR20240010707A (ko) 2024-01-24
ZA202300601B (en) 2024-06-26

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