CN114703097A - Bacillus and application thereof, and multi-strain fermentation composite fermentation plant enzyme - Google Patents
Bacillus and application thereof, and multi-strain fermentation composite fermentation plant enzyme Download PDFInfo
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- CN114703097A CN114703097A CN202210349257.2A CN202210349257A CN114703097A CN 114703097 A CN114703097 A CN 114703097A CN 202210349257 A CN202210349257 A CN 202210349257A CN 114703097 A CN114703097 A CN 114703097A
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- 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
Abstract
The invention belongs to the technical field of biology, and particularly relates to bacillus and application thereof, and a multi-strain fermentation composite fermentation plant enzyme. The invention provides two specific bacillus strains, application thereof and a multi-strain fermentation composite fermentation plant enzyme. The plant enzyme provided by the invention can effectively utilize the overground part of the rhubarb, the prepared plant enzyme can effectively improve the commodity characters of the cultivated plants, and meanwhile, the preparation method of the plant enzyme provided by the invention is simple and has extremely high marketization prospect.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to bacillus and application thereof, and a multi-strain fermentation composite fermentation plant enzyme.
Background
Rhubarb, also known as rheum officinale, saussurea involucrate, fresh isatis root, Ladingwen name Rumex madaio Makino. Rumex species (Rumex) plants Rumex patoria (r. patientia), Rumex japonicus (r. japonica), Rumex acetosa (r. acetosa), Rumex palustris (r. obtusifolia L), Rumex nepalensis (r. nepalensis) and Rumex crispatus (r.gmelinii) are collectively called Rumex madaio. Rheum officinale originates from the slope of the original wild mountain and is widely distributed in regions of China.
Due to the important medicinal value of the rhubarb, the rhubarb has the characteristics of cold resistance, drought resistance, saline-alkali resistance, barren resistance and sand-barren resistance, and is suitable for planting in various soils, and large-area planting is developed in many areas. Particularly, the growth amount of Rumex madaio leaves is much larger than the yield of roots of Rumex madaio leaves, and the yield per mu of Rumex madaio leaves in northern years is 20-40 tons; the southern annual acre yield is 35-60 tons. Meanwhile, the rhubarb is a perennial herb, but the medicinal rhubarb is only harvested once by using the underground part, if the leaf harvesting is mainly used for planting, the planting yield is very considerable, repeated planting is not needed for years or even decades once, and the rhubarb can be continuously harvested. However, although the root of rhubarb is mainly used in the research and use of medicine at present, the leaves of rhubarb are non-toxic and edible, but the leaves are not popular with the public because of sour taste, so the overground part of rhubarb is difficult to utilize, and the utilization value of the leaves of rhubarb is in need of development.
Disclosure of Invention
In order to solve the problems, the invention provides two specific bacillus strains, application thereof and a multi-strain fermentation composite fermentation plant enzyme. The plant enzyme provided by the invention can effectively utilize the overground part of the rhubarb, the prepared plant enzyme can effectively improve the commodity characters of the cultivated plants, and meanwhile, the preparation method of the plant enzyme provided by the invention is simple and has extremely high marketization prospect.
In order to achieve the purpose, the invention provides the following technical scheme:
the invention provides a Zuante province bacillus, which is preserved in China general microbiological culture Collection center with the preservation number of CGMCC No. 22667;
the bacillus aryabhattai is preserved in China general microbiological culture collection center with the preservation number of CGMCC No. 22669.
The invention also provides application of the bacillus zungterii and/or bacillus aryabhattai in preparation of plant enzymes.
The invention provides a plant ferment, which comprises the following raw materials: a fermentation product, a leavening agent and a bacterial liquid;
the fermentation product comprises rhubarb root, a carbon source, fungus leftovers and water;
the bacterial liquid comprises the bacillus driller province and the bacillus aryabhattai.
Preferably, the fermented product of the plant ferment comprises the following components in parts by mass: 20-25 parts of rheum officinale, 5-7 parts of a carbon source, 7-10 parts of fungus leftovers and 96-126 parts of water;
the using mass of the leavening agent is 0.03 percent of the total mass of the fermentation system;
the using mass of the bacterial liquid is 0.02% of the total mass of the fermentation system;
the concentration of the bacterial liquid is 1 multiplied by 108cfu/mL, wherein the ratio of Bacillus thuringiensis to Bacillus aryabhattai in the bacterial liquid is 1: 1.
preferably, the leavening agent comprises lactic acid bacteria and/or yeast.
Preferably, the Rumex madaio includes Rumex patientia, Rumex japonicus, Rumex acetosa, Rumex obtusifolius, Rumex nepalensis and Rumex madaio.
The invention also provides a preparation method of the plant enzyme, which comprises the following steps: and mixing the fermentation product, the leavening agent and the bacterial liquid, and fermenting for 28-31 days.
Preferably, the temperature of the fermentation is 20-25 ℃.
The invention provides application of the plant enzyme or the plant enzyme prepared by the preparation method in promoting plant growth.
Preferably, the plant for promoting plant growth comprises capsicum.
The invention provides a bacillus zungteri and bacillus aryabhattai and application thereof in preparation of plant enzymes. The bacillus thuringiensis provided by the invention has the functions of producing cellulase, protease and amylase, and can promote the fermentation of raw materials to produce enzymes; the Bacillus aryabhattai has a good degradation effect on organic matters such as crop straws and herbicides, and particularly has a good degradation effect on atrazine in saline-alkali soil with high salt content; both of the above-mentioned bacilli have biocontrol potential for inhibiting the growth of harmful microorganisms in soil. Consequently, add these two kinds of bacilli and carry out the ferment preparation, not only can promote the fermentation effect of raw materials, can strengthen the beneficial function of ferment simultaneously.
According to the plant enzyme provided by the invention, the yeast, the lactic acid bacteria and the beneficial agricultural microorganisms are added to ferment the leaves of the rhubarb and the fungus leftovers to prepare the high-quality agricultural enzyme, so that the waste of plant resources can be effectively avoided, the agricultural pollution of the Jianshan can be realized, and the resource utilization of agricultural waste materials can be realized. The Rumex madaio leaf used in the invention can convert rich crude protein, total phosphorus, total sugar, vitamins, beta-carotene, chlorophyll, emodin, isoflavone, SOD, selenium, potassium, calcium, iron, zinc, phosphorus, magnesium and other microelements and beneficial minerals into enzyme solution with rich sugar, organic acid and mineral components through the fermentation effect of microorganisms. The ferment liquid is used in crop production, can increase soil nutrient substances, improve soil structure, promote nutrient absorption, further improve crop physiological indexes, enhance plant resistance, reduce crop pest and disease damage, and realize 'fertilizer and pesticide dual reduction' and 'dual increase' of yield and quality in crop production. In the invention, the fungus leftovers are preferably pleurotus eryngii leftovers, because the yield of the pleurotus eryngii is extremely high and the material source is rich, the pleurotus eryngii is generally produced in a factory, a large amount of leftovers are produced after the pleurotus eryngii is harvested and treated every day, and the leftovers contain a large amount of nutrient substances such as polysaccharide and the like and are similar to the nutrient components of the fruiting body of the pleurotus eryngii. Therefore, the leftovers of the pleurotus eryngii are also high-quality raw materials for preparing the ferment. In the embodiment of the invention, the pleurotus eryngii leftovers can be cut (the particle size is less than 1cm) or pulped for use.
The examples show that the effect of the plant ferment can be effectively improved by using the bacillus zungterii and bacillus aryabhattai provided by the invention. The plant enzyme provided by the invention can effectively improve the plant height, fruit bearing number and fruit commodity properties of cultivated plants, and after the plant enzyme provided by the invention is used, the average plant height of the cayenne pepper can be improved by 15.96%, the average fruit bearing number is improved by 28.57%, the diameter of the fruit is increased by 6.25%, the fruit length is improved by 8.29%, the weight of the fruit is increased by 36.51%, and the fruit length, the glossiness and the like are also obviously increased.
Proof of deposit
Bacillus drenchensis (Bacillus drenchensis) preserved in China scientific microbiology institute No. 3 of Xilu No. 1 of Beijing, Chaoyang, on 2021, 08 days 06 and with the preservation number of CGMCC No. 22667;
bacillus aryabhattai (Bacillus aryabhattai) is deposited at China scientific microbiology institute, No. 3 of Xilu No. 1 of Beijing, Chaoyang, on 2021, 06 months 08, with the deposit number of CGMCC No. 22669.
Drawings
FIG. 1 shows the growth of a plant enzyme treated pepper plant according to the present invention and a control pepper plant;
FIG. 2 is a comparison of plant enzyme treated pepper fruits of the present invention and control pepper fruits.
Detailed Description
The reagents and instruments used in the following description are not specifically described in terms of techniques or conditions, and are performed under the conditions of the conventional experiments, or under the conditions suggested in the specification. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.
Example 1
1. Material
7 parts of carbon source (brown sugar and/or sucrose, in the embodiment of the invention, brown sugar is selected), 10 parts of pleurotus eryngii leftovers, 20-25 parts of fresh rhubarb leaves (rumex bambusicola, 100 parts of water, a leaven and a bacterial liquid.
2. Material processing
Carbon source: mixing conventional commercially available brown sugar with appropriate amount of water (distilled water is used in the examples of the present invention), and heating to melt;
the leftovers of pleurotus eryngii: cleaning leftovers such as mushroom feet and mushroom pieces from a pleurotus eryngii factory production field, cutting the leftovers into pieces (smaller than 1cm) or directly beating the pieces into pulp for later use;
fresh rhubarb leaves: directly cutting the harvested rumex patientia from the microbiological research institute of Guangxi agricultural academy into pieces (< 1cm) for later use;
and (3) a leavening agent: the leaven is lactobacillus and yeast purchased from Angel yeast flagship; the mass ratio of the lactic acid bacteria to the yeast is 3: 1; the using amount of the leavening agent is 0.03 percent of the total mass of the fermentation system;
bacterial liquid: respectively and independently activating and culturing Bacillus thuringiensis (Bacillus drenchensis) with the preservation number of CGMCC No.22667 and Bacillus aryabhattai with the preservation number of CGMCC No.22669 by LB culture media (indoor normal-temperature fermentation, the initial pH value is 3.8, and the pH value is about 3.0 after one month of fermentation), and centrifugally collecting thalli; the cells were resuspended in sterile water to a concentration of 1X 108cfu/mL of bacterial suspension (the ratio of Bacillus thuringiensis to Bacillus aryabhattai in the bacterial suspension is 1: 1); the adding amount of the bacterial liquid during fermentation is 0.02 percent of the total mass of the fermentation system.
3. Fermentation process
Mixing the processed raw materials in proportion, uniformly mixing and stirring the raw materials, transferring the raw materials to a common enzyme fermentation tank (a barrel-shaped container with a sealing cover), sealing and fermenting the raw materials at the temperature of between 20 and 25 ℃, and fermenting the raw materials for 1 month to finish the preparation of the plant enzyme provided by the invention.
4. Applications of
Effect of the application of the plant ferment provided by the invention on the growth of pepper (the test planting is linear pepper)
And (3) transplanting the pepper (linear pepper, the same below) when seedling is grown to 4-5 true leaves, fertilizing according to 1/3 of the recommended fertilizing amount for production, and managing the rest conditions according to the conventional planting requirements.
The experimental group applies 100 times of diluent of the plant enzyme provided by the invention to each of the seedling stage and the initial flowering stage of the pepper, the dosage of the diluent is 100mL for each plant in the seedling stage and 150mL for each plant in the initial flowering stage, and the diluent is applied by root irrigation; the control group was applied with an equal amount of clear water.
When the pepper grows to the initial stage of fruiting, spraying 150 times of diluent of the plant enzyme provided by the invention on the leaf surface of the plant, wherein the dosage of the diluent is 45 liters of the enzyme diluent sprayed per mu; the control group was sprayed with equal amount of clear water.
After the peppers are transplanted for 70 days, the following indexes are measured when the first peppers are mature: plant height, fruit bearing number of a single pepper plant, fruit length, fruit thickness and average single fruit weight, and calculating the weight of all fruits; the growth vigor of the plants, the fruit glossiness and the like are observed and photographed. Results are shown in fig. 1, fig. 2 and table 1, wherein fig. 1 is a comparison of growth vigor of plant enzyme treated litsea cubeba plants provided by the present invention and control litsea cubeba plants, and fig. 2 is a comparison of plant enzyme treated litsea cubeba fruits provided by the present invention and control litsea cubeba fruits.
TABLE 1 Effect of plant enzymes on Pepper growth
As can be seen from fig. 1, fig. 2 and table 1, the plant height, the fruit number per plant, the fruit length, the fruit thickness and the fruit weight of the pepper treated with the plant enzyme provided by the present invention are respectively increased by 15.96%, 28.57%, 6.25%, 8.29% and 36.51% compared with the control, which indicates that the plant enzyme provided by the present invention has an excellent plant growth promoting effect.
The foregoing is only a preferred embodiment of the invention and it should be noted that modifications and embellishments could be made by the inventor without departing from the principle of the invention and should be considered as the scope of the invention.
Claims (10)
1. Bacillus thuringiensis (Bacillus drenchensensis) preserved in China general microbiological culture collection center with the preservation number of CGMCC No. 22667;
bacillus aryabhattai (Bacillus aryabhattai) is preserved in China general microbiological culture collection center with the preservation number of CGMCC No. 22669.
2. Use of bacillus macerans and/or bacillus aryabhattai according to claim 1 for the preparation of plant ferments.
3. The plant ferment is characterized in that the plant ferment comprises the following raw materials: a fermentation product, a leavening agent and a bacterial liquid;
the fermentation product comprises rhubarb root, a carbon source, fungus leftovers and water;
the carbon source comprises brown sugar and/or sucrose;
the bacterial liquid comprises the bacillus thuringiensis and bacillus aryabhattai of claim 1.
4. The plant ferment of claim 3, wherein the fermented product of the plant ferment comprises the following components in parts by mass: 20-25 parts of rheum officinale, 5-7 parts of a carbon source, 7-10 parts of fungus leftovers and 96-126 parts of water;
the using mass of the leavening agent is 0.03 percent of the total mass of the fermentation system;
the using mass of the bacterial liquid is 0.02% of the total mass of the fermentation system;
the concentration of the bacterial liquid is 1 multiplied by 108cfu/mL, wherein the ratio of Bacillus thuringiensis to Bacillus aryabhattai in the bacterial liquid is 1: 1.
5. the plant ferment of claim 3, wherein the leavening agent comprises lactic acid bacteria and/or yeast.
6. The plant ferment of claim 3, wherein the dock root comprises Rumex patientia, Rumex japonicus, Rumex acetosa, Rumex obtusifolius, Rumex nepalensis, and Rumex gmelini.
7. The method for preparing the plant ferment of any one of claims 3 to 6, wherein the method comprises the following steps: and mixing the fermentation product, the leavening agent and the bacterial liquid, and fermenting for 28-31 days.
8. The method according to claim 7, wherein the fermentation temperature is 20 to 25 ℃.
9. Use of the plant ferment of any one of claims 3 to 6 or the plant ferment obtained by the preparation method of claim 7 or 8 in promoting plant growth.
10. The use as claimed in claim 9, wherein the plant growth promoting plant comprises capsicum.
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