NL2030811B1 - PLANT GROWTH-PROMOTING RHIZOBACTERIA BACILLUS SP. Lzh-5 AND USE THEREOF - Google Patents

PLANT GROWTH-PROMOTING RHIZOBACTERIA BACILLUS SP. Lzh-5 AND USE THEREOF Download PDF

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NL2030811B1
NL2030811B1 NL2030811A NL2030811A NL2030811B1 NL 2030811 B1 NL2030811 B1 NL 2030811B1 NL 2030811 A NL2030811 A NL 2030811A NL 2030811 A NL2030811 A NL 2030811A NL 2030811 B1 NL2030811 B1 NL 2030811B1
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lzh
mulberry wine
preparation
mulberry
bacillus
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Zhu Feng
Wang Zheng
Zhao Li
Wang Jintao
Li Fangting
Du Shaoguo
Li Zhenghua
Zhao Zhe
ran Kun
Xin Yuxiu
Zhan Bin
Sun Xiaoli
Song Feng
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Univ Dezhou
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    • 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
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F11/00—Other organic fertilisers
    • C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F5/00—Fertilisers from distillery wastes, molasses, vinasses, sugar plant or similar wastes or residues, e.g. from waste originating from industrial processing of raw material of agricultural origin or derived products thereof
    • 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
    • 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
    • C12R2001/07—Bacillus

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  • General Health & Medical Sciences (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Botany (AREA)
  • Environmental & Geological Engineering (AREA)
  • Fertilizers (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The present disclosure relates to a plant growth-promoting rhizobacteria (PGPR) Bacillus sp. Lzh-S and use thereof. The PGPR Bacillus sp. Lzh-S was deposited at the China General Microbiological Culture Collection Center (CGMCC), NO. 1 West Beichen Road, Chaoyang District, Beijing 100101, China on December 23, 2016, with an accession number of CGMCC No. 13487. The present disclosure further includes use of the PGPR Bacillus sp. Lzh-S in a microbial fertilizer. Not only can the present disclosure promote plant root growth effectively and prevent the occurrence of root rot, but substantially improve a utilization rate of mulberry wine stillages and reduce environmental pollution.

Description

PLANT GROWTH-PROMOTING RHIZOBACTERIA BACILLUS SP. Lzh-5 AND
USE THEREOF
TECHNICAL FIELD
[OI] The present disclosure relates to a plant growth-promoting rhizobacteria (PGPR)
Bacillus sp. Lzh-5 and use thereof, and belongs to the field of agricultural product processing and microbial fertilizer preparation.
BACKGROUND ART
[02] Plant rhizosphere soil contains a large number of plant growth-promoting rhizobacteria (PGPR), and many species of PGPR have been found, such as Bacillus sp.,
Brevibacillus sp, Paenibacillus sp, Acinetobacter sp, Agrobacterium sp., and
Streptomyces sp. Among them, Bacillius sp. is one of the most deeply and widely studied
PGPR due to its spore production and strong resistance.
[03] PGPR is inoculated into an organic adsorption carrier to prepare a microbial organic fertilizer. Due to low production cost, microbial organic fertilizers can significantly improve the quality of agricultural products and reduce the application of chemical fertilizers, playing an increasingly important role in agricultural production and having an excellent application prospect. At present, turf is the most common microbial organic fertilizer carrier on the market, but the turf is expensive, so it is necessary to look for a new microbial fertilizer carrier.
[04] Mulberry wine stillage is the main by-product produced during the brewing process of mulberry wine. Although the pigment and flavonoids thereof have been extracted, the separation effect is low and a large amount of waste residues are still discarded after separation, resulting in environmental pollution and huge waste of resources. Therefore, there is an urgent need to look for a way to make full use of the mulberry wine stillage. The present disclosure provides a method for extracting anthocyanins from the mulberry wine stillage by using ultrasonic assisted technology, thereby substantially improving the extraction rate of anthocyanins. The mulberry wine stillage is also rich in trace elements and minerals, which is of great development value and can be used as a carrier for microbial organic fertilizer.
SUMMARY
[05] An objective of the present disclosure is to provide a PGPR Bacillus sp. Lzh-5 and use thereof, and the strain of the present disclosure is a strain having significant antagonistic activity isolated from a field from Dezhou City, Shandong Province. The present disclosure can be used in a microbial fertilizer. Through a simple preparation method, not only can the microbial fertilizer promote plant root growth effectively and prevent the occurrence of root rot, but substantially improve a utilization rate of mulberry wine stillages and reduce environmental pollution. The technical solutions of the present disclosure are as follows:
[06] A PGPR Bacillus sp. Lzh-5 is provided, deposited at the China General
Microbiological Culture Collection Center (CGMCC), NO. 1 West Beichen Road,
Chaoyang District, Beijing 100101, China on December 23, 2016, with an accession number of CGMCC No. 13487. The taxonomic denomination is Bacillus sp.
[07] The present disclosure further includes use of the PGPR Bacillus sp. Lzh-5 in the preparation of a microbial fertilizer.
[08] A microbial fertilizer is provided, including a mulberry wine stillage after anthocyanin extraction, an organic fertilizer, and a fermentation broth of a PGPR Bacillus sp. Lzh-5.
[09] The present disclosure provides a preparation method of the microbial fertilizer, including the following steps:
[19] step 1, preparation of the mulberry wine stillage after anthocyanin extraction:
[11] drying the mulberry wine stillage in an air dry oven at 50°C for 24 h to remove volatile acids, pulverizing the mulberry wine stillage to obtain a mulberry wine stillage powder, sealing, and storing at 4°C in the dark; mixing 100-200 g of the mulberry wine stillage powder with 1 L of distilled water for sonication, where sonication conditions are: temperature 55-65°C, ultrasonic power 380-420 W, and time 80-100 min; and conducting vacuum filtration, where a resulting filter residue is the mulberry wine stillage after anthocyanin extraction; preferably, a pulverized mulberry wine stillage powder has a particle size of 45-60 mesh; the sonication is conducted at a temperature of 60°C and an ultrasonic power of 400 W for 90 min;
[12] step 2, preparation of the fermentation broth of the PGPR Bacillus sp. Lzh-5
[13] inoculating an inoculum of the PGPR Bacillus sp. Lzh-5 with inoculum size of 2% (v/v) into a Luria-Bertani (LB) broth having a pH of 7.0, and fermenting at 25-30°C and 180-250 rpm for 45-50 h to obtain the fermentation broth, where preferably, the inoculation is followed by fermentation at 28°C and 200 rpm for 48 h;
[14] step 3, preparation of the microbial fertilizer
[15] mixing the mulberry wine stillage after anthocyanin extraction obtained in step 1 with the organic fertilizer at a mass ratio of (1-3): (3-9); after mixing, adding 0.1 mL/g resulting mixture into the fermentation broth of the strain Lzh-5 obtained in step 2, stirring evenly, and conducting a second fermentation at 30°C for 72 h to obtain a microbial fertilizer containing the mulberry wine stillage, where preferably, the mulberry wine stillage after anthocyanin extraction and the organic fertilizer are mixed at a mass ratio of 3:7.
[16] Further, the organic fertilizer may be a swine manure compost.
[17] Compared with the prior art, the present disclosure has the following advantages:
[18] (1) The PGPR Bacillus sp. Lzh-5 has a strong antagonistic effect on the pathogenic fungus Fusarium moniliforme;
[19] (2) The mulberry wine stillage is rich in trace elements such as potassium, calcium and molybdenum, and minerals, which can promote the growth of seedlings;
[20] (3) The microbial fertilizer of the present disclosure can promote plant root growth effectively and prevent the occurrence of root rot.
BRIEF DESCRIPTION OF THE DRAWINGS
[21] FIG. 1 illustrates an antagonistic effect of strain Lzh-5 on root rot pathogen
Fusarium moniliforme Sheld, where FIG. 1A illustrates a confrontation culture experiment of the strain Lzh-5 against the root rot pathogen + moniliforme Sheld, and FIG. 1B illustrates the growth of control Fusarium solani.
[22] FIG. 2 is a pot experiment for a promoting effect of the microbial fertilizer of the present disclosure on plant growth, where control CK represents no application of organic fertilizer in the soil infected by root rot pathogen, control CK2 represents application of only organic fertilizer in the soil infected by the root rot pathogen, treatment groups (1) to (5) represent application of microbial fertilizers obtained in Examples 2 to 6 in the soil infected by root rot pathogen.
DETAILED DESCRIPTION OF THE EMBODIMENTS
[23] The present disclosure will be further described below in conjunction with specific examples, and the advantages and features of the present disclosure will become clearer from the description. However, the examples are merely exemplary and do not constitute any limitation on the scope of the present disclosure. Those skilled in the art will appreciate that modifications and substitutions of the technical solutions of the present disclosure can be made in form and detail without departing from the spirit and scope of the present disclosure, but all of these modifications and substitutions fall within the protection scope of the present disclosure.
[24] Example 1 Identification of the strain Lzh-5
[25] The strain Lzh-5 was a strain having significant antagonistic activity isolated from a field from Dezhou City, Shandong Province. The strain Lzh-5 was identified by morphological observation, staining, and 16S rRNA gene sequence analysis. The results showed that: the strain Lzh-5 was a Gram-positive bacterium, and it was a milky white round colony on the LB agar plate, which was opaque and flat, irregular, moist on the surface, and easy to pick. After 16S rRNA gene sequence analysis and homology comparison, it had 99% homology with a Bacillus strain in GenBank, so it was identified as Bacillus sp, named Bacillus sp. Lzh-5.
[26] The strain Lzh-5 was deposited at the China General Microbiological Culture
Collection Center (CGMCC). 1 West Beichen Road, Chaoyang District, Beijing 100101,
China on December 23, 2016, with an accession number of CGMCC No. 13487. The accession number of the 16S rRNA gene of this strain in GenBank is KX865134.
[27] Antagonistic activity: The pathogenic fungus F. moniliforme Sheld was inoculated onto Potato Dextrose Agar (PDA, 200 g of peeled potato, 20 g of sucrose, 20 g of agar, 1,000 mL of distilled water, at natural pH), and cultured at 30°C for 48 h for later use. The strain L.zh-5 was inoculated into LB agar (10 g of peptone, 5 g of yeast extract, 10 g of
NaCl, 20 g of agar, 1,000 mL of distilled water, at pH = 7.4-7.6), cultured at 30°C for 48 h, inoculated on both sides of the pathogenic fungus, and continued to culture at 30°C for 48-72 h, followed by observing whether an inhibition zone was produced. The results are shown in FIG. 1. The results showed that the strain Lzh-5 of the present disclosure had a strong antagonistic effect.
[28] Example 2 A microbial fertilizer and a preparation method thereof
[29] The preparation steps of the microbial fertilizer were as follows:
[30] Step 1, preparation of a mulberry wine stillage after anthocyanin extraction:
[31] The mulberry wine stillage was dried in an air dry oven at S0°C for 24 h to remove volatile acids, pulverized to obtain a mulberry wine stillage powder, sealed, and stored at
4°C in the dark; 100-200 g of the mulberry wine stillage powder was mixed with 1 L of distilled water for sonication, where sonication conditions were: temperature 60°C, ultrasonic power 400 W, and time 90 min; and vacuum filtration was conducted, where a resulting filter residue was the mulberry wine stillage after anthocyanin extraction; the 5 pulverized mulberry wine stillage powder was 50 mesh in particle size.
[32] Step 2, preparation of a fermentation broth of a PGPR Bacillus sp. Lzh-5
[33] An inoculum of the PGPR Bacillus sp. Lzh-5 with inoculum size of 2% (v/v) was inoculated into an LB broth having a pH of 7.0, and fermented at 28°C and 200 rpm for 48 h to obtain the fermentation broth.
[34] Step 3, preparation of a microbial fertilizer
[35] The mulberry wine stillage after anthocyanin extraction obtained in step 1 was mixed with an organic fertilizer, swine manure compost, at a mass ratio of 3:7, after mixing, 0.1 mL/g resulting mixture was added into the fermentation broth of the strain
Lzh-5 obtained in step 2, stirred evenly, and subjected to a second fermentation at 30°C for 72 h to obtain a microbial fertilizer containing the mulberry wine stillage.
[36] Example 3 A microbial fertilizer and a preparation method thereof
[37] The preparation steps of the microbial fertilizer were as follows:
[38] Step 1, preparation of a mulberry wine stillage after anthocyanin extraction:
[39] The mulberry wine stillage was dried in an air dry oven at 50°C for 24 h to remove volatile acids, pulverized to obtain a mulberry wine stillage powder, sealed, and stored at 4°C in the dark; 100-200 g of the mulberry wine stillage powder was mixed with 1 L of distilled water for sonication, where sonication conditions were: temperature 55°C, ultrasonic power 380 W, and time 100 min; and vacuum filtration was conducted, where a resulting filter residue was the mulberry wine stillage after anthocyanin extraction; the pulverized mulberry wine stillage powder was 60 mesh in particle size.
[40] Step 2, preparation of a fermentation broth of a PGPR Bacillus sp. Lzh-5
[41] An inoculum of the PGPR Bacillus sp. Lzh-5 with inoculum size of 2% (v/v) was inoculated into an LB broth having a pH of 7.0, and fermented at 25°C and 230 rpm for 50 h to obtain the fermentation broth.
[42] Step 3, preparation of a microbial fertilizer
[43] The mulberry wine stillage after anthocyanin extraction obtained in step 1 was mixed with an organic fertilizer, swine manure compost, at a mass ratio of 2:3; after mixing, 0.1 mL/g resulting mixture was added into the fermentation broth of the strain
Lzh-5 obtained in step 2, stirred evenly, and subjected to a second fermentation at 30°C for 72 h to obtain a microbial fertilizer containing the mulberry wine stillage.
[44] Example 4 A microbial fertilizer and a preparation method thereof
[45] The preparation steps of the microbial fertilizer were as follows:
[46] Step 1, preparation of a mulberry wine stillage after anthocyanin extraction:
[47] The mulberry wine stillage was dried in an air dry oven at 50°C for 24 h to remove volatile acids, pulverized to obtain a mulberry wine stillage powder, sealed, and stored at 4°C in the dark; 100-200 g of the mulberry wine stillage powder was mixed with 1 L of distilled water for sonication, where sonication conditions were: temperature 65°C, ultrasonic power 410 W, and time 80 min; and vacuum filtration was conducted, where a resulting filter residue was the mulberry wine stillage after anthocyanin extraction; the pulverized mulberry wine stillage powder was 45 mesh in particle size.
[48] Step 2, preparation of a fermentation broth of a PGPR Bacillus sp. Lzh-5
[49] An inoculum of the PGPR Bacillus sp. Lzh-5 with inoculum size of 2% (v/v) was inoculated into an LB broth having a pH of 7.0, and fermented at 29°C and 190 rpm for 45 h to obtain the fermentation broth.
[50] Step 3, preparation of a microbial fertilizer
[51] The mulberry wine stillage after anthocyanin extraction obtained in step 1 was mixed with an organic fertilizer, swine manure compost, at a mass ratio of 1:4; after mixing, 0.1 mL/g resulting mixture was added into the fermentation broth of the strain
Lzh-5 obtained in step 2, stirred evenly, and subjected to a second fermentation at 30°C for 72 h to obtain a microbial fertilizer containing the mulberry wine stillage. [S2] Example 5 A microbial fertilizer and a preparation method thereof
[53] The preparation steps of the microbial fertilizer were as follows:
[54] Step L, preparation of a mulberry wine stillage after anthocyanin extraction:
[55] The mulberry wine stillage was dried in an air dry oven at 50°C for 24 h to remove volatile acids, pulverized to obtain a mulberry wine stillage powder, sealed, and stored at 4°C in the dark; 100-200 g of the mulberry wine stillage powder was mixed with 1 L of distilled water for sonication, where sonication conditions were: temperature 58°C, ultrasonic power 420 W, and time 80 min; and vacuum filtration was conducted, where a resulting filter residue was the mulberry wine stillage after anthocyanin extraction; the pulverized mulberry wine stillage powder was 50 mesh in particle size.
[56] Step 2, preparation of a fermentation broth of a PGPR Bacillus sp. Lzh-5
[S7] An inoculum of the PGPR Bacillus sp. Lzh-5 with inoculum size of 2% (v/v) was inoculated into an LB broth having a pH of 7.0, and fermented at 26°C and 210 rpm for 47 h to obtain the fermentation broth. [S8] Step 3, preparation of a microbial fertilizer
[59] The mulberry wine stillage after anthocyanin extraction obtained in step 1 was mixed with an organic fertilizer, swine manure compost, at a mass ratio of 1:1; after mixing, 0.1 mL/g resulting mixture was added into the fermentation broth of the strain
Lzh-5 obtained in step 2, stirred evenly, and subjected to a second fermentation at 30°C for 72 h to obtain a microbial fertilizer containing the mulberry wine stillage.
[60] Example 6 A microbial fertilizer and a preparation method thereof
[61] The preparation steps of the microbial fertilizer were as follows:
[62] Step 1, preparation of a mulberry wine stillage after anthocyanin extraction:
[63] The mulberry wine stillage was dried in an air dry oven at 50°C for 24 h to remove volatile acids, pulverized to obtain a mulberry wine stillage powder, sealed, and stored at 154°C in the dark; 100-200 g of the mulberry wine stillage powder was mixed with 1 L of distilled water for sonication, where sonication conditions were: temperature 63°C, ultrasonic power 380 W, and time 95 min; and vacuum filtration was conducted, where a resulting filter residue was the mulberry wine stillage after anthocyanin extraction; the pulverized mulberry wine stillage powder was 50 mesh in particle size.
[64] Step 2, preparation of a fermentation broth of a PGPR Bacillus sp. Lzh-5
[65] An inoculum of the PGPR Bacillus sp. Lzh-5 with inoculum size of 2% (v/v) was inoculated into an LB broth having a pH of 7.0, and fermented at 25°C and 230 rpm for 48 h to obtain the fermentation broth.
[66] Step 3, preparation of a microbial fertilizer
[67] The mulberry wine stillage after anthocyanin extraction obtained in step 1 was mixed with an organic fertilizer, swine manure compost, at a mass ratio of 1:9; after mixing, 0.1 mL/g resulting mixture was added into the fermentation broth of the strain
Lzh-5 obtained in step 2, stirred evenly, and subjected to a second fermentation at 30°C for 72 h to obtain a microbial fertilizer containing the mulberry wine stillage.
[68] Test Example Pot experiment
[69] Test species: pepper seedlings
[70] Experimental treatments:
[71] Control 1: no organic fertilizer was applied to the soil infected by the root rot pathogen
[72] Control 2: only organic fertilizer was applied to the soil infected by the root rot pathogen
[73] Treatment (1): the microbial fertilizer obtained in Example 2 was applied to the soil infected by the root rot pathogen
[74] Treatment (2): the microbial fertilizer obtained in Example 3 was applied to the soil infected by the root rot pathogen
[75] Treatment (3): the microbial fertilizer obtained in Example 4 was applied to the soil infected by the root rot pathogen
[76] Treatment (4): the microbial fertilizer obtained in Example 5 was applied to the soil infected by the root rot pathogen
[77] Treatment (5): the microbial fertilizer obtained in Example 6 was applied to the soil infected by the root rot pathogen
[78] In the pot experiment, for each treatment, the fertilizers of each test group were applied to 10 pepper seedlings with uniform growth. After the soil and fertilizer were mixed, the seedlings were covered with soil and watered, and other measures were managed as usual. After the plants grew, the growth length of the root system was measured and calculated, and the growth status and disease indexes of the plants were investigated and recorded, and the control effect was calculated.
[79] The experimental results are shown in Table 1 and FIG. 2. Pepper seedlings applied with the microbial fertilizer prepared by mixing the mulberry wine stillage and organic fertilizer at a ratio of 3:7 grew fastest with developed root systems. Moreover, the addition of microbial fertilizer could significantly increase the growth rate of the plant and reduce the incidence.
[80] The experimental results show that the microbial fertilizer of the present disclosure can effectively promote the plant root growth and control the incidence of root rot.
[81] Table 1 Effect of the microbial fertilizer of the present disclosure on the root length of pepper seedlings and its control effect on root rot

Claims (8)

Conclusies l. Gebruik van een plantengroeibevorderende rhizobacterie (PGPR) Bacillus sp. Lzh-5 in een microbiéle meststof.Conclusions l. Use of a plant growth-promoting rhizobacterium (PGPR) Bacillus sp. Lzh-5 in a microbial fertilizer. 2. Microbiéle meststof, waarbij de microbiéle meststof een moerbeiwijnvinasse na anthocyanine-extractie, een organische meststof en een fermentatiebouillon van een PGPR Bacillus sp. Lzh-5 omvat.2. A microbial fertilizer, wherein the microbial fertilizer comprises a mulberry wine vinasse after anthocyanin extraction, an organic fertilizer and a fermentation broth of a PGPR Bacillus sp. Lzh-5 includes. 3. Bereidingswerkwijze van de microbiële meststof volgens conclusie 2, waarbij de bereidingswerkwijze de volgende stappen omvat: stap 1, het bereiden van de moerbeiwijnvinasse na anthocyanine-extractie: het drogen van de moerbeiwijnvinasse in een luchtdroogoven bij 50 °C gedurende 24 uur om vluchtige zuren te verwijderen, het verpulveren van de moerbeiwijnvinasse om een moerbeiwijnvinassepoeder te verkrijgen, het afdichten en het opslaan bij 4 °C in het donker; het mengen van 100 — 200 g van het moerbeiwijnvinassepoeder met 1 L gedestilleerd water voor sonificatie, waarbij sonificatieomstandigheden de volgende zijn: temperatuur 55 — 65 °C, ultrasoon vermogen 380 — 420 W en tijd 80 — 100 min; en het uitvoeren van vacuümfiltratie, waarbij een resulterend filterresidu de moerbeiwijnvinasse na anthocyanine-extractie is; stap 2, het bereiden van de fermentatiebouillon van de PGPR Bacillus sp. Lzh-5: het inoculeren van een inoculum van de PGPR Bacillus sp. Lzh-5 met inoculumgrootte van 2 volume-% in een Luria-Bertani- (LB-) bouillon met een pH van 7,0 en het fermenteren bij 25 — 30 °C en 180 — 250 rpm gedurende 45 — 50 uur om de fermentatiebouillon te verkrijgen; stap 3, het bereiden van de microbiële meststof: het met de organische meststof mengen van de moerbeiwijnvinasse na anthocyanine-extractie die verkregen is in stap 1 in een massaverhouding van (1 -3):G3 — 9); het toevoegen van 0,1 mL/g resulterend mengsel in de fermentatiebouillon van de stam Lzh-5 die verkregen is in stap 2, het gelijkmatig roeren en het uitvoeren van een tweede fermentatie bij 30 °C gedurende 72 uur om een microbiële meststof te verkrijgen die de moerbeiwijnvinasse omvat.The preparation method of the microbial fertilizer according to claim 2, wherein the preparation method comprises the following steps: step 1, preparing the mulberry vinasse after anthocyanin extraction: drying the mulberry vinasse in an air-drying oven at 50°C for 24 hours to remove volatile acids to remove, crushing the mulberry wine vinasse to obtain a mulberry wine vinasse powder, sealing and storing at 4°C in the dark; mixing 100 - 200 g of the mulberry vinasse powder with 1 L of distilled water for sonication, where sonication conditions are as follows: temperature 55 - 65 °C, ultrasonic power 380 - 420 W and time 80 - 100 min; and performing vacuum filtration, a resulting filter residue being the mulberry wine vinasse after anthocyanin extraction; step 2, preparing the fermentation broth of the PGPR Bacillus sp. Lzh-5: inoculating an inoculum of the PGPR Bacillus sp. Lzh-5 with inoculum size of 2% by volume in a Luria-Bertani (LB) broth with a pH of 7.0 and fermenting at 25-30°C and 180-250 rpm for 45-50 hours to obtain the fermentation broth to obtain; step 3, preparing the microbial fertilizer: mixing with the organic fertilizer the mulberry wine vinasse after anthocyanin extraction obtained in step 1 in a mass ratio of (1 -3):G3 - 9); adding 0.1 mL/g of the resulting mixture into the fermentation broth of the Lzh-5 strain obtained in step 2, stirring uniformly, and conducting a second fermentation at 30 °C for 72 hours to obtain a microbial fertilizer which includes the mulberry wine vinasse. S11 -S11 - 4. Bereidingswerkwijze volgens conclusie 3, waarbij een verpulverd moerbeiwijnvinassepoeder in stap 1 een deeltjesgrootte van 45 — 60 maas heeft.The preparation method according to claim 3, wherein a pulverized mulberry wine vinasse powder in step 1 has a particle size of 45-60 mesh. 5. Bereidingswerkwijze volgens conclusie 3, waarbij de sonificatie in stap 1 uitgevoerd wordt bij een temperatuur van 60 °C en een ultrasoon vermogen van 400 W gedurende 90 min.The preparation method according to claim 3, wherein the sonication in step 1 is carried out at a temperature of 60°C and an ultrasonic power of 400 W for 90 min. 6. Bereidingswerkwijze volgens conclusie 3, waarbij de inoculatie in stap 2 gevolgd wordt door fermentatie bij 28 °C en 200 rpm gedurende 48 uur.The preparation method according to claim 3, wherein the inoculation in step 2 is followed by fermentation at 28°C and 200 rpm for 48 hours. 7. Bereidingswerkwijze volgens conclusie 3, waarbij in stap 3 de moerbeiwijnvinasse na anthocyanine-extractie en de organische meststof gemengd worden in een massaverhouding van 3:7.The preparation method according to claim 3, wherein in step 3 the mulberry wine vinasse after anthocyanin extraction and the organic fertilizer are mixed in a mass ratio of 3:7. 8. Bereidingswerkwijze volgens conclusie 3, waarbij de organische meststof in stap 3 een zwijnenmestcompost is.The preparation method according to claim 3, wherein the organic fertilizer in step 3 is a boar manure compost.
NL2030811A 2022-02-03 2022-02-03 PLANT GROWTH-PROMOTING RHIZOBACTERIA BACILLUS SP. Lzh-5 AND USE THEREOF NL2030811B1 (en)

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