NL2031759B1 - Bacillus and its Application and Multi-strain Fermentation Compound Fermentation Plant Enzyme - Google Patents
Bacillus and its Application and Multi-strain Fermentation Compound Fermentation Plant Enzyme Download PDFInfo
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- 102000004190 Enzymes Human genes 0.000 title claims abstract description 58
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 58
- 238000000855 fermentation Methods 0.000 title claims abstract description 39
- 230000004151 fermentation Effects 0.000 title claims abstract description 39
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- 238000002360 preparation method Methods 0.000 claims description 13
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- NCYCYZXNIZJOKI-IOUUIBBYSA-N 11-cis-retinal Chemical compound O=C/C=C(\C)/C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C NCYCYZXNIZJOKI-IOUUIBBYSA-N 0.000 description 1
- VZSRBBMJRBPUNF-UHFFFAOYSA-N 2-(2,3-dihydro-1H-inden-2-ylamino)-N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]pyrimidine-5-carboxamide Chemical compound C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)C(=O)NCCC(N1CC2=C(CC1)NN=N2)=O VZSRBBMJRBPUNF-UHFFFAOYSA-N 0.000 description 1
- 102100040190 ADP-ribosylation factor-binding protein GGA2 Human genes 0.000 description 1
- 235000001674 Agaricus brunnescens Nutrition 0.000 description 1
- 239000004382 Amylase Substances 0.000 description 1
- 102000013142 Amylases Human genes 0.000 description 1
- 108010065511 Amylases Proteins 0.000 description 1
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- 108010059892 Cellulase Proteins 0.000 description 1
- 241000723363 Clerodendrum Species 0.000 description 1
- 235000019750 Crude protein Nutrition 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 101001037082 Homo sapiens ADP-ribosylation factor-binding protein GGA2 Proteins 0.000 description 1
- UPYKUZBSLRQECL-UKMVMLAPSA-N Lycopene Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1C(=C)CCCC1(C)C)C=CC=C(/C)C=CC2C(=C)CCCC2(C)C UPYKUZBSLRQECL-UKMVMLAPSA-N 0.000 description 1
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- 241000219050 Polygonaceae Species 0.000 description 1
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- 235000019418 amylase Nutrition 0.000 description 1
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- CJWQYWQDLBZGPD-UHFFFAOYSA-N isoflavone Natural products C1=C(OC)C(OC)=CC(OC)=C1C1=COC2=C(C=CC(C)(C)O3)C3=C(OC)C=C2C1=O CJWQYWQDLBZGPD-UHFFFAOYSA-N 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
-
- 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
- A01N63/22—Bacillus
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- 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
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- 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
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
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Abstract
The invention relates to the technical field of biotechnology, in particularly to bacillus and its application and multi-strain fermentation compound fermentation plant enzyme; the present invention provides two specific bacilli and their applications and multi-strain fermentation compound fermentation plant enzyme; the plant enzyme provided by the present invention can effectively utilize the above-ground part of RumeX madaio Makino, and the plant enzyme made from it can effectively improve the commercial traits of cultivated plants. At the same time, the plant enzyme provided by the present invention is simple to prepare and has a high prospect of marketability.
Description
Bacillus and its Application and Multi-strain Fermentation Compound
Fermentation Plant Enzyme
The invention relates to the technical field of biotechnology, in particularly to bacillus and its application and multi-strain fermentation compound fermentation plant enzyme.
Rumex madaio Making IR. darwoo Makino], also known as fresh manytiower glorybower root, is a genus of Polygonum in the family Polygonaceae, The Rumex plants
R. panentig, R. japomcs, R. acetosa, R. obtusifohus LL, BR. vepalensis, R. gioeling, ete. are collectively known as Rumex madaio Makino. Rumex madain Makino grows on the slopes of wilderness, and is widely distributed nationwide in China,
Rumex madaio Makino has an important medicinal value and is resistant to cold, drought, salinity, barrenness and sandy sail, which makes it suitable for planting in many areas. Therefore, it has been planted on a large scale in many areas. In particular, the yield of Kumex madain Makino leaves is much larger than that of its roots, the annual yield of
Rurnex madaro Makino nthe north is about 20 to 40 tons per 10u per vear, and its yield in the south is about 35 to 60 tons per mu per year. Meanwhile, Rumex madaio Makino is a perepmal herb, However, Rumex madato Makino is used only for the undergronnd part and can only be harvested once. If the main purpose of planting is to harvest the leaves, us planting yield 1s considerable, and it can be planted once for several years or even decades without re-planting but continuous substantial harvests are available
However the roots of Rumex madaio Makino are mainly used for medical research and nse at present. Although the leaves are non-toxic and edible, they are vot popular with the public because otf their sour taste. Therefore, the above-ground part of Rumex mada
Makino is difficult to be utilized, and the value of the leaves of Rumex madaio Makino needs ic be developed.
The purpose of the invention is to overcome the above problems. The present invention provides two specific bacilli and their applications and multi-strain fermentation compound fermentation plant enzyme. The plant enzyme provided by the present invention can effectively utilize the above-ground part of Rumex madaio Makino, and the plant enzyme made from it can effectively improve the commercial traits of cultivated plants. At the same time, the plant enzyme provided by the present invention is simple to prepare and has a high prospect of marketability.
To achieve the above purpose, the present invention provides the following solutions:
The present invention provides a bacillus drentensis, and the bacillus drentensis is deposited in the China General Microbiological Culture Collection Center under the deposit number CGMCC No. 22667; a bacillus aryabhattai and the bacillus aryabhattai deposited in the China General
Microbiological Culture Collection Center under the deposit number CGMCC No.22669.
The present invention also provides the application of the bacillus drentensis and/or bacillus aryabhattai in the preparation of plant enzymes.
The present invention provides a plant enzyme, and the raw materials of the plant enzyme include leavening, fermentation agent and bacteria solution;
The leavening includes Rumex madaio Makino, carbon source, fungal leftovers and water;
The bacteria solution includes bacillus drentensis and bacillus aryabhattai.
The further improvement is that the leavening in the plant enzyme includes the following components by mass: 20-25 parts of Rumex madaio Makino, 5-7 parts of carbon source, 7-10 parts of fungal leftovers and 96-126 parts of water;
The mass of the fermentation agent used is 0.03% of the total mass of the fermentation system;
The mass of the bacteria solution used is 0.02% of the total mass of the fermentation system;
The concentration of the bacteria solution is 1x 103 cfu/mL, and the ratio of bacillus drentensis and bacillus aryabhattai in the bacteria solution is 1: 1.
The further improvement is that the fermentation agent includes lactic acid bacteria and / yeast.
The further improvement is that the Rumex madaio Makino includes R.patientia,
R.japonics, R.acetosa, R.obtusifolius L, R.nepalensis and R.gmelinii.
The present invention also provides the preparation method of the plant enzyme, and the preparation method includes the following steps: mixing the leavening, fermentation agent and bacteria solution and fermenting them for 28~31 days.
The further improvement is that the temperature of the fermentation is 20°C to 25°C.
The present invention provides the application of plant enzymes obtained from the plant enzymes or the preparation methods in the promotion of plant growth.
The further improvement is that the plants that promote plant growth include capsicum.
The present invention provides a bacillus drentensis and a bacillus aryabhattai and their applications in the preparation of plant enzymes. The bacillus drentensis provided by the present invention has the function of cellulase, protease and amylase production, and it can promote the fermentation of raw materials to produce enzymes; bacillus aryabhattai has a good degradation effect on organic materials such as crop straw and herbicides, and especially has excellent degradation effects on atrazine in saline soil with high salt content; the two categories of bacilli have the biocontrol potential to inhibit the growth of hazardous microorganisms in the soil. Therefore, the addition of these two bacilli for enzyme preparation can not only improve the fermentation effect of raw materials, but also enhance the beneficial functions of the enzyme.
By adding yeast, lactic acid bacteria and the above mentioned beneficial agricultural microorganisms, the plant enzyme provided by the present invention ferments the leaves of Rumex madaio Makino and the fungus leftovers to prepare high-quality agricultural enzymes, so that it can effectively avoid the waste of plant resources, reduce agricultural pollution, and realize the resource utilization of agricultural waste materials. The leaves of Rumex madaio Makino used in the present invention are fermented by microorganisms to transform the rich crude protein, total phosphorus, total sugar, vitamins, as well as 3- carotene, chlorophyll, rhodopsin, isoflavones, SOD, selenium, potassium, calcium, iron, zinc, phosphorus, magnesium and other trace elements and beneficial minerals in the leaves into an enzyme liquid with rich sugar, organic acids and mineral components.
Enzyme liquid used in crop production can increase soil nutrients, improve soil structure, and promote nutrient absorption. In turn, it can improve crop physiological indicators, enhance plant resistance, reduce crop diseases and pests, and achieve "double reduction of fertilizers and pesticides" and "double increase of yield and quality" in crop production.
In the present invention, the fungal leftovers are preferably pleurotus eryngii leftovers. The yield of pleurotus eryngii is very large, and the source of material is abundant, and pleurotus eryngii is generally produced in factories. After daily harvesting and processing, a large amount of leftovers are produced, and leftovers contain a large amount of polysaccharides and other nutrients, which are similar to the nutritional composition of the fruiting bodies of pleurotus eryngii. Therefore, the leftovers of
Pleurotus eryngii are also a good raw material for enzyme preparation. In the embodiments of the present invention, the leftovers of pleurotus eryngii can be chopped (particle size <lcm) or pulpized.
It can be seen from the embodiments that the use of bacillus drentensis and bacillus aryabhattai provided by the present invention can effectively improve the effect of plant enzymes. The plant enzyme provided by the present invention can effectively improve the plant height, the number of fruits and the commercial properties of fruits of cultivated plants. After the use of the plant enzyme provided by the present invention, the average plant height of capsicum annuum var. longunt can increase by 15.96%, the average number of fruits can increase by 28.57%, the diameter of fruits can increase by 6.25%, the fruit length can increase by 8.29%, and the weight of 100 fruits can increase by 36.51%. Moreover, there 1s a significant increase in fruit length and gloss.
DEPOSITORY CERTIFICATE
The bacillus drentensis was deposited at No.1 Beichen West Road, Chaoyang
District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, on June 8, 2021, under the deposit number CGMCC No.22667,
The bacillus aryabhattai was deposited at No.1 Beichen West Road, Chaoyang 5 District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, on June 8, 2021, under the deposit number CGMCC No.22669.
Fig. 1 is the comparison of the growth of capsicum annuum var. longunt plants treated with the plant enzyme provided by the present invention and the capsicum annuum var. longunt plants of the control group;
Fig. 2 is the comparison of the fruits of capsicum annuum var. longunt plants treated with the plant enzyme provided by the present invention and the fruits of the capsicum annuum var. longunt plants of the control group.
If the reagents or instruments used in the following paragraphs do not specify specific techniques or conditions, they will be performed under conventional experimental conditions; if the instructions of the reagent company are not specified, they will be performed under the conditions suggested by the instructions. The reagents and instruments used without specifying the manufacturer are conventional products that can be purchased from the market.
Example 1 1. Materials 7 parts of carbon source (brown sugar and/sucrose, brown sugar chosen in the embodiment of the present invention), 10 parts of pleurotus eryngii leftovers, 20-25 parts of fresh Rumex madaio Makino leaves (R. patientia), 100 parts of water, fermentation agent and bacteria solution. 2. Material processing
Carbon source: Mixing conventional commercially available brown sugar with an appropriate amount of water (distilled water 1s used in the embodiment of the present invention) and then heating and melting it;
Pleurotus eryngii leftovers: Removinh impurities from mushroom trunks and slices and other pleurotus eryngii leftovers sourced from pleurotus eryngii factory production sites, chopping them (less than 1 cm in diameter) or just smashing them into a paste;
Fresh Rumex madaio Makino leaves: Collecting R patientia from the experimental field of the Institute of Microbiology, Guangxi Academy of Agricultural Sciences, chopping them (less than 1 cm in diameter) for future use;
Fermentation agent: The fermentation agent is the lactic acid bacteria and yeast are purchased from Angel Yeast flagship store; the mass ratio of lactic acid bacteria to yeast is 3: 1; the mass of the fermentation agent used is 0.03% of the total mass of the fermentation system;
Bacteria solution: Activating bacillus drentensis (CGMCC No. 22667) and bacillus aryabhattai (CGMCC No. 22669) independently in LB medium ( fermentation at room temperature, pH 3.8 at the beginning and 3.8 at the end of the fermentation for one month) and collecting the bacteria by centrifugation; resuspending the bacteria in sterile water to make a suspension with a concentration of 1X 108 cfu/mL (the ratio of bacillus drentensis to bacillus aryabhattai in the suspension is 1:1); the mass of the bacteria solution added is 0.02% of the total mass of the fermentation system during fermentation. 3. Fermentation process
Mixing the processed raw materials in proportion, transferring the well-mixed raw materials to a common fermentation tank (barrel-shaped container with a sealed lid) and fermenting them in a sealed environment at 20°C~25°C for | month to complete the preparation of the plant enzyme provided by the present invention. 4. Application
Effect of application of plant enzymes provided by the present invention on the growth of capsicum (capsicum annuum var. longunt planted in the experiment)
The seedlings of capsicum (capsicum annuum var. longunt, hereafter the same) are transplanted only when they have 4-5 main leaves and are fertilized at 1/3 of the recommended fertilizer dosage for production and the rest of the conditions are managed according to the conventional planting requirements.
In the treatment group, a 100-fold dilution of plant enzymes provided by the present invention is applied at the seedling stage and early flowering stage of capsicum at the rate of 100 mL per plant at the seedling stage and 150 mL per plant at the early flowering stage, applying by root irrigation; the control group is applied with the same amount of water.
In the early stage of capsicum fruiting, the leaf surfaces of the plants are sprayed with 150 times dilution of plant enzymes provided by the present invention at a dosage of 45 liters of enzyme dilution per mu. The control group is sprayed with the same amount of water.
The following indicators are measured 70 days after capsicum transplanting, when the first capsicums are ripe: plant height, number of fruits per plant, fruit length, fruit thickness, average fruit weight and the weight of 100 fruits; plant growth and fruit gloss are observed and photographed. The results are shown in Fig. 1., Fig. 2 and Tab. 1. Fig. 1 is the comparison of the growth of capsicum annuum var. longunt plants treated with the plant enzyme provided by the present invention and the capsicum annuum var. longunt plants of the control group; Fig. 2 is the comparison of the fruits of capsicum annuum var. longunt plants treated with the plant enzyme provided by the present invention and the fruits of the capsicum annuum var. longunt plants of the control group.
Tab. 1 Effect of plant enzyme on the growth of capsicum e || cme
Treatment Control group enzymes
Ear [eee EE»
Plant height 38 | 49 33 plant (cm) 3 4 6 5 8 5 2 3 $
Mean of the treatment 56.35 48.77 group (cm)
Increase compared to 15.96 the control group (%)
Mean per 15116 {1110} 12 9 9 | 13 | 9 | 10 plant (fruit)
Mean of the treatment 12
Number of group (fruit) fruits
Increase compared to the control group (%)
Mean per 53 52150 50149151150 50146 plant (cm) 6 7 8 1 1 3 4
Mean of the treatment 5.16 4.86
Fruit group (cm) thickness
Increase compared to the control group (%)
Mean per 22.121. 20. 15. 23. | 20. | 21. | 19. 20. 22 22 plant (cm) 4 3 5 2 3 8 9 4 6
Fruit length
Mean of the 21.9 treatment
Increase compared to 8.29 the control group (%)
Mean per 21 17 | 21 17 119 15 | 23 | 20 | 21 | 19 | 20 #4 plant(g) | 350 | 32 | 14 | 70 | 64 20 | 46 | 80 | 90 | 40 | 55
Mean of the treatment 1965 1439.5
Weight of group (g) 100 fruits
Increase compared to 36.51 the control group (%)
According to Fig.1, Fig. 2 and Tab. 1, the plant height, number of fruits per plant, fruit length, fruit thickness and weight of 100 fruits of capsicum treated with plant enzymes provided by the present invention increase by 15.96%, 28.57%, 6.25%, 8.29% and 36.51% respectively compared with the control group. It shows that the plant enzyme provided by the present invention has an excellent effect of promoting plant growth.
The above embodiments only describe the preferred mode of the invention, but do not limit the scope of the invention. On the premise of not departing from the design spirit of the invention, various modifications and improvements made by the inventor of the present invention in the field to the technical scheme of the invention shall fall within the protection scope determined by the claims of the invention.
Claims (10)
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