CN110622996B - Microbial agent and application thereof - Google Patents
Microbial agent and application thereof Download PDFInfo
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- CN110622996B CN110622996B CN201910917894.3A CN201910917894A CN110622996B CN 110622996 B CN110622996 B CN 110622996B CN 201910917894 A CN201910917894 A CN 201910917894A CN 110622996 B CN110622996 B CN 110622996B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- 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
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Abstract
The invention provides a microbial agent and application thereof, and relates to the technical field of biological agents, wherein the microbial agent comprises the following components in parts by weight: citric acid fermentation tail liquid, peptone, yeast powder, alkalophilic bacillus and water; the volume of the citric acid fermentation tail liquid, the alkalophilic bacillus and the water, and the mass ratio of the peptone to the yeast powder are (15-25) mL, (2-3) mL, (975-985) mL, (8-12) g and (4-6) g; the content of organic matters in the citric acid fermentation tail liquid is 150-250 g/L. The microbial agent provided by the invention not only has an obvious effect of improving the soil of saline-alkali soil, but also can promote the growth of crops, increase the yield of the crops and improve the quality of fruits.
Description
Technical Field
The invention relates to the technical field of biological agents, and particularly relates to a microbial agent and application thereof.
Background
Citric acid is widely used as an important chemical raw material and a food additive in the industries of food, medicine, chemical industry and the like. At present, China has nearly one hundred families of citric acid production plants, the total annual production capacity is about 80 ten thousand tons, and the citric acid production plants are the biggest citric acid production country and export country in the world. The citric acid tail liquid mainly contains starch, protein, carbohydrate, various organic acids, ammonia nitrogen, fat and the like, the concentration is 20000-30000 mg/L, and the citric acid tail liquid belongs to high-concentration organic wastewater, and if the high-concentration organic wastewater is directly discharged, the environment is polluted, and the soil is damaged (Wangming, Liao building, high-concentration organic wastewater biological treatment technology [ J ]. contemporary chemical industry, 2011,40, (8) 820-. The citric acid treatment method includes a physical treatment method, a chemical treatment method and a biological treatment method. The biological treatment method comprises the steps of putting a microbial agent or a membrane-fixed flora to convert ammonia nitrogen, inorganic salt and nitrite nitrogen in water or sediment, transferring decomposition products to algae, continuously supplying dissolved oxygen generated by algae photosynthesis to aerobic flora, and mutually utilizing the dissolved oxygen to mutually promote a large amount of waste water organisms to be absorbed and utilized to achieve the effect of purifying waste water. Some researchers mix the citric acid waste water with other starch waste water according to a certain proportion to ferment Bacillus, as microbial fertilizer, promote plant growth, improve soil environment (C.Zhou, S.xu, M.Zhang et al.Utilization of sweet potato starch water for bio-organic fertilizer [ M ]. Sustainable Irrisation and Draainage IV: 237-.
The salinization of soil is a worldwide problem, China is one of the world saline-alkali land big countries, and the total area of the salinized soil is about 0.99 multiplied by 108hm2(Zhujianfeng, treble, Wuchunhong, etc. the research progress and prospect of saline-alkali soil greening in China [ J]The world forestry research, 2018,31(04):70-75.), and large-area saline-alkali soil not only causes huge economic loss on agricultural production, but also threatens local ecological environment. For a long time, saline-alkali soil can be improved by using improvement measures such as irrigation and leaching, covering salt control, chemical modifier application and the like, the growth of plants is promoted, but the utilization cost is high, and secondary pollution is easy to generate.
Disclosure of Invention
In view of the above, the present invention aims to provide a microbial agent and an application thereof. The microbial agent provided by the invention not only has an obvious effect of improving the soil of saline-alkali soil, but also can promote the growth of crops, increase the yield of the crops and improve the quality of fruits.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a microbial agent, which comprises: citric acid fermentation tail liquid, peptone, yeast powder, alkalophilic bacillus and water; the volume of the citric acid fermentation tail liquid, the alkalophilic bacillus and the water, and the mass ratio of the peptone to the yeast powder are (15-25) mL, (2-3) mL, (975-985) mL, (8-12) g and (4-6) g;
the content of organic matters in the citric acid fermentation tail liquid is 150-250 g/L.
Preferably, the total viable count of the microbial agent is not less than2×108CFU/mL。
Preferably, the alkalophilic bacillus comprises wp-1, and the deposit number of the wp-1 is CCTCC NO: m2018427.
Preferably, the pH value of the microbial agent is 5.5-6.5.
Preferably, the solution for adjusting the pH value of the microbial agent comprises ammonia water, and the mass concentration of the ammonia water is 20-30%.
The invention also provides application of the microbial agent in the technical scheme in improvement of saline-alkali soil.
The invention also provides the function of the microbial agent in the technical scheme in promoting plant growth.
The invention also provides application of the microbial agent in the technical scheme in improving plant yield.
The invention also provides application of the microbial agent in the technical scheme in improving the quality of plants.
Preferably, the plants include lycium barbarum, wheat, and tomato.
The invention provides a microbial agent and application thereof. The microbial agent provided by the invention can effectively utilize citric acid fermentation tail liquid, promote plant growth, increase yield, improve quality and improve saline-alkali soil.
In the embodiment of the invention, the microbial agent provided by the invention has a promotion effect on growth of new shoots and rhizome of Chinese wolfberry, and is increased by 15% compared with a control; the content of soluble solid matters of the medlar, the vitamin C, the polysaccharide, the titratable acid and the carotenoid is respectively increased by 13.22 percent, 46.38 percent, 16.13 percent, 46.88 percent and 63.02 percent compared with the conventional field fertilization; the yield per mu of fresh fruits is increased by 5.22 percent compared with the fertilizer application of the conventional field, and the yield per mu of dry fruits is increased by 12.11 percent.
In the embodiment of the invention, the application of the microbial agent provided by the invention has a promoting effect on soil improvement of various soil layers, and has a better improvement effect compared with a commercial microbial agent and conventional field fertilization; the soil pH, conductivity and total salt content of each soil layer are all reduced compared with CK, the soil quick-acting potassium and quick-acting phosphorus are all improved, and the change of the organic matter content in the soil layer of 20-40cm is not obvious.
Drawings
FIG. 1 is a graph showing the effect of the microbial agents of the present invention on the growth of young shoots of Lycium barbarum;
FIG. 2 is a graph showing the effect of the microbial agents of the present invention on the growth of Lycium barbarum stems.
Detailed Description
The invention provides a microbial agent, which comprises: citric acid fermentation tail liquid, peptone, yeast powder, alkalophilic bacillus and water; the volume of the citric acid fermentation tail liquid, the alkalophilic bacillus and the water, and the mass ratio of the peptone to the yeast powder are (15-25) mL, (2-3) mL, (975-985) mL, (8-12) g and (4-6) g; the content of organic matters in the citric acid fermentation tail liquid is 150-250 g/L.
The source of the citric acid fermentation tail liquid is not particularly limited, and the citric acid fermentation tail liquid obtained by producing citric acid by adopting a fermentation method in the field can be obtained. In the invention, the organic matter content of the citric acid fermentation tail liquid is 150-250 g/L, preferably 180-220 g/L, and more preferably 200 g/L. In the embodiment of the invention, the pH value of the citric acid tail liquid is 3.35, the organic matter content is 200.55g/L, the humic acid content is 2.35g/L, the total nitrogen content is 106.10g/L, the total phosphorus content is 2.82g/L and the total potassium content is 16.85 g/L.
The sources of the peptone and the yeast powder are not particularly limited, and the peptone and the yeast powder can be obtained by adopting conventional commercial products.
In the present invention, the water is preferably deionized water.
In the invention, the alkaliphilic Bacillus is preferably alkaliphilic Bacillus (Bacillus pseudofirmus) wp-1, the wp-1 is preserved in 2018, 7 and 2 months and is preserved in China center for type culture Collection, wherein the China center for type culture Collection in Wuchang Lojia mountain Wuhan university school is provided, the strain preservation number is CCTCC NO: m2018427.
In the invention, the volume of the citric acid fermentation tail liquid, the alkalophilic bacillus, the water and the mass ratio of the peptone to the yeast powder are (15-25) mL, (2-3) mL, (975-985) mL, (8-12) g and (4-6) g; preferably 20mL, 2.5mL, 980mL, 10g, 5 g.
In the present invention, in the case of the present invention,the viable count of the alkalophilic bacillus in the microbial agent is preferably more than or equal to 2 multiplied by 108CFU/mL。
In the present invention, the preparation method of the microbial agent preferably includes: mixing the citric acid fermentation tail liquid, water, peptone and yeast powder, adjusting the pH to 5.5-6.5 by using ammonia water, and sterilizing at 115-125 ℃ for 15-25 min to obtain a mixed solution; and inoculating the alkalophilic bacillus into the mixed solution for culturing to obtain the microbial agent.
In the present invention, the mass concentration of the ammonia water is preferably 20 to 30%, and more preferably 25%. The pH value is preferably 5.5-6.5, and more preferably 6.0. The invention adopts ammonia water to adjust the pH value because other alkaline solutions such as sodium hydroxide, potassium hydroxide and the like can bring potassium and sodium ions into the soil, thereby increasing the saline-alkali degree of the soil.
In the invention, the sterilization temperature is preferably 115-125 ℃, and more preferably 121 ℃; the time for sterilization is preferably 15-25 min, and more preferably 20 min.
In the invention, the temperature of the culture is preferably 25-35 ℃, and more preferably 30 ℃; the time for culturing is preferably 2-3 d.
The invention also provides application of the microbial agent in the technical scheme in improvement of saline-alkali soil.
The invention also provides the function of the microbial agent in the technical scheme in promoting plant growth.
The invention also provides application of the microbial agent in the technical scheme in improving plant yield.
The invention also provides application of the microbial agent in the technical scheme in improving the quality of plants.
In the present invention, the plant preferably includes lycium barbarum, wheat, and tomato.
In the present invention, the mode of application of the microbial agent is not particularly limited, and a hole application method is preferably used. In the present invention, the application amount of the microbial agent is applied according to the fertilizer requirement of crops. In the embodiment of the invention, the application amount of the microbial agent is preferably 15-25 mL/(hole/strain), and more preferably 20 mL/(hole/strain).
The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
Preparation method of microbial agent
Diluting 20mL of citric acid fermentation tail liquid to 1000mL by 980mL of distilled water, adding 10g of peptone and 5g of yeast powder into the liquid at room temperature, stirring uniformly, and adjusting the pH value to 6.0 by using 25% ammonia water. The liquid is subpackaged into 1000mL glass triangular bottles to ensure that the liquid amount in the bottles is 500mL, sealed by a sealing film, sterilized at 121 ℃ for 20min, and the sterilized liquid is placed at room temperature. Wherein the pH value of the citric acid tail liquid is 3.35, the organic matter content is 200.55g/L, the humic acid content is 2.35g/L, the total nitrogen content is 106.10g/L, the total phosphorus content is 2.82g/L and the total potassium content is 16.85 g/L.
Bacillus alkalophilus wp-1 strain in LB slant culture medium stored in a refrigerator at 4 deg.C was inoculated into sterilized LB liquid culture medium of pH8.0, and shaken at 30 deg.C and 180r/min for 48 h. Wherein the preservation number of the bacillus alcalophilus wp-1 strain is CCTCC NO: m2018427. The LB liquid culture medium is prepared by mixing 5g of yeast powder, 10g of peptone, 10g of sodium chloride and 1000mL of distilled water, subpackaging and sterilizing at 121 ℃ for 20 min.
And (3) putting the bacillus alcalophilus wp-1 cultured for 48 hours into a 50mL centrifuge tube, centrifuging for 2min (4 ℃, 10000r/min) by using a high-speed centrifuge, diluting the obtained thallus by using a 1mL pipette gun with a sterilization liquid containing the citric acid fermentation tail liquid, adding 2mL of bacterial liquid into the sterilized liquid containing the citric acid fermentation tail liquid, mixing, fully shaking uniformly, and placing in a 30 ℃ shaking table 2d at 180r/min to obtain the microbial agent. Wherein the total viable count of the microbial agent is 2 multiplied by 108CFU/mL。
Example 2
The microbial agent of example 1 is used for growth, quality, yield and improvement of saline-alkali soil of medlar.
Test zone 90m2The medlar field which is grown for 3 years, has uniform density and consistent growth vigor is used as a test land, and the medlar variety is Ningqi No. 7. The test adopts a completely random test design, the test is provided with T1 treatment (conventional fertilization + commercial microbial inoculum), T2 treatment (conventional fertilization + microbial inoculum of example 1) and control treatment CK (conventional fertilization), the treatment and the control are repeated for 3 times, and 5 trees with uniform growth vigor are selected in each treatment. Digging 20cm deep holes 20cm away from the tree body in three directions by a hole application method, and applying fertilizers for 6 times from the early tip-pulling period of 5 months to the middle fruitage period of 7 months in 2019, wherein each fertilizer is applied by 20ml per plant. The field management is the same as the conventional field management. The soil conditions in the test area are shown in table 1, and belong to alkaline and severe salinized soil.
TABLE 1 soil background value of 0-20cm soil layer in test area
Measuring the growth amount of the new shoots of the medlar:
and selecting the observation sample plants and the observation branches in late 4 months in 2019. 3 sample plants with similar tree vigor are selected in each treatment, 1 new observation branch is respectively selected and reserved in the middle of the crown according to five directions of east, south, west, north and middle, the observation branches are marked by red wool, and the observation plants are not trimmed during the whole growth period. Measuring once every 7 days from 4 late ten days to 6 late ten days, and measuring by a measuring tape. The results are shown in FIG. 1. The application of the microbial inoculum can promote the growth of new shoots of the medlar, and the treatment with T2 is most obvious, and is increased by 15 percent compared with the conventional fertilization.
And (3) measuring the stem growth amount of the medlar:
the plants are measured to be the same as the young sprout growth amount plants, and are measured once every 7 days from 4 to 6 late months, and the measurement is carried out by a vernier caliper. The results are shown in FIG. 2. The application of the microbial inoculum has a promoting effect on the growth of the stems of the Chinese wolfberry, and the treatment with T2 is most obvious, and is increased by 15 percent compared with the conventional fertilization.
And (3) determining the quality of the medlar:
fruits are taken in the full bearing period, stored in a foam box with an ice bag and taken back to the laboratory as soon as possible, one part is stored at low temperature (4 ℃) for measuring the internal quality, and the other part is naturally aired for measuring the yield of the dried fruits. The method comprises the steps of measuring soluble solid solids by a handheld method, measuring the carotenoid content of fresh fruits by GB 12291-90, measuring the vitamin C content by a 2, 6-dichloroindophenol titration method, measuring the polysaccharide content by an anthrone sulfuric acid method, and measuring titratable acid by an acid-base neutralization titration method. The weight was weighed on a hundredth scale to obtain the dry fruit mass (g/strain). The results are shown in tables 2 and 3.
TABLE 2 influence of the application of the inoculum on the medicinal quality of Lycium barbarum
The content of soluble solid, polysaccharide and titratable acid of the medlar are important indexes for determining the taste and flavor of the fruits, and the carotenoid and the polysaccharide are known as important internal medicine components of the medlar and have the effects of enhancing immunity, resisting cancer and resisting aging. Vitamin C is not only a nutrient substance essential to human bodies, but also an important index for resisting aging and stress in the storage process of fruits and vegetables. As shown in the table 2, the quality content of the medlar fruits can be improved by applying the microbial inoculum, the treatment performance is obvious by applying the microbial inoculum, and the content of medlar soluble solids, vitamin C, polysaccharide, titratable acid and carotenoid is respectively increased by 13.22%, 46.38%, 16.13%, 46.88% and 63.02% compared with the conventional field fertilization.
TABLE 3 Effect of application of the inoculum on wolfberry production
The fertilization is an important way for increasing the agricultural production and the income of farmers. As shown in Table 2, the yield per mu of the product microbial inoculum is improved compared with that of the commercial microbial inoculum and that of the conventional field fertilization. The yield per mu of fresh fruits is increased by 5.22 percent compared with the fertilizer application of the conventional field, and the yield per mu of dry fruits is increased by 12.11 percent; the yield per mu of fresh fruits is increased by 4.85 percent compared with the treatment of commercial microbial inoculum.
Determination of soil improvement:
soil in soil layers of 0-20cm and 20-40cm in three directions and 20cm away from the tree body in the full bearing period are respectively and fully mixed, repeated for three times, filled into a sealing bag and taken back to the room for soil nutrition analysis. The pH of the soil adopts a potential method; the conductivity adopts a water-soil ratio 5:1 method; drying the whole salt by adopting a residue drying-mass method; organic matters adopt a potassium dichromate-external heating method; the quick-acting phosphorus adopts a molybdenum-antimony colorimetric method; the quick-acting potassium is obtained by flame photometry. The results are shown in Table 4.
TABLE 4 Effect of application of the inoculum on soil improvement
As shown in Table 4, the microbial inoculum has a promoting effect on soil improvement of various soil layers after being applied, and the microbial inoculum has a better improvement effect than commercial microbial inocula and conventional field fertilization. In a soil layer of 0-20cm, the pH value of the soil is reduced by 0.30 unit at most compared with that of CK after the treatment of T2, the conductivity of the soil is reduced by 8.65%, the total salt content of the soil is reduced by 1.52%, and organic matters, quick-acting potassium and quick-acting phosphorus are respectively improved by 32.08%, 19.57% and 118.54% compared with CK. In a soil layer of 20-40cm, the pH value, the conductivity and the total salt of the soil are all reduced compared with CK, the quick-acting potassium and the quick-acting phosphorus of the soil are all improved, and the change of the organic matter content is not obvious.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (8)
1. A microbial inoculant comprising: citric acid fermentation tail liquid, peptone, yeast powder, alkalophilic bacillus and water; the volume of the citric acid fermentation tail liquid, the alkalophilic bacillus and the water, and the mass ratio of the peptone to the yeast powder are (15-25) mL, (2-3) mL, (975-985) mL, (8-12) g and (4-6) g;
the content of organic matters in the citric acid fermentation tail liquid is 150-250 g/L;
the Bacillus alcalophilus is wp-1, and the preservation number of the wp-1 is CCTCC NO: m2018427, and the number of viable bacteria is more than or equal to 2 multiplied by 108CFU/mL。
2. The microbial agent according to claim 1, wherein the pH value of the microbial agent is 5.5-6.5.
3. The microbial agent according to claim 2, wherein the solution for adjusting the pH value of the microbial agent comprises ammonia water, and the mass concentration of the ammonia water is 20-30%.
4. The application of the microbial agent as claimed in any one of claims 1 to 3 in improving saline-alkali soil.
5. Use of the microbial inoculant according to any one of claims 1 to 3 for promoting plant growth.
6. Use of the microbial inoculant of any one of claims 1-3 for increasing plant yield.
7. Use of the microbial inoculant according to any one of claims 1 to 3 for improving plant quality.
8. Use according to any one of claims 5 to 7, wherein the plant comprises Lycium barbarum, wheat and tomato.
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