CN112385507B - Seedling culture medium containing arbuscular mycorrhiza and preparation method and application thereof - Google Patents
Seedling culture medium containing arbuscular mycorrhiza and preparation method and application thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/05—Fruit crops, e.g. strawberries, tomatoes or cucumbers
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
- A01G24/12—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material containing soil minerals
- A01G24/15—Calcined rock, e.g. perlite, vermiculite or clay aggregates
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/50—Growth substrates; Culture media; Apparatus or methods therefor contained within a flexible envelope
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05D—INORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C; FERTILISERS PRODUCING CARBON DIOXIDE
- C05D9/00—Other inorganic fertilisers
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
- C05F17/20—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation using specific microorganisms or substances, e.g. enzymes, for activating or stimulating the treatment
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
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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
- C12N1/14—Fungi; Culture media therefor
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
Abstract
The invention relates to a seedling substrate containing arbuscular mycorrhiza and a preparation method and application thereof. The substrate of the invention consists of an organic substrate, an inorganic substrate and arbuscular mycorrhizal fungi strains. The method is characterized in that the crop seedling raising substrate contains arbuscular mycorrhizal fungi, and the arbuscular mycorrhizal fungi are encapsulated. The encapsulation of the capsule can separate the direct contact of the arbuscular mycorrhizal fungi and the matrix, avoid the inhibition of the nutrients in the matrix, especially the quick-acting phosphorus, on the germination of the arbuscular mycorrhizal strains, keep the strain activity and prolong the shelf life. The capsule can controllably dissolve and release the encapsulated arbuscular mycorrhizal fungi to form an arbuscular mycorrhizal structure with the crop root system. The dissolution time of the capsule can be regulated by adjusting the water content of the matrix, the capsule can be completely dissolved in 8 hours under the condition of 100 percent of water content, and the dissolution time is prolonged along with the reduction of the water content of the matrix.
Description
Technical Field
The invention belongs to the technical field of crop seedling culture and particularly relates to a seedling culture substrate containing arbuscular mycorrhiza and a preparation method and application thereof.
Background
With the rapid development of facility gardening, organic substrate cultivation becomes a new soilless cultivation technology in greenhouses, and the research on organic substrates is promoted. The organic matter resource is very rich, and can be local organic matters such as crop straws, carbonized rice hulls, wheat hulls, mushroom dregs, grass carbon mud, grass carbon, coconut shells, barks, sawdust, wood shavings, sunflower straws, bagasse, vinasse, corncobs, tobacco dregs, flax, waste cotton, wine dregs, fruit dreg sawdust, reed powder, waste paper pulp, traditional Chinese medicine dregs, bean dregs, oil meal (rapeseed cakes, cotton cakes, bean cakes and the like), municipal garbage and the like; the livestock and poultry manure can also become a better organic matrix material through fermentation or high-temperature treatment. However, the existing organic matrix has hard texture, inconvenient operation, poor air permeability, poor water retention, poor nutrient conversion efficiency and poor seedling culture effect, so that the matrix which has loose texture and good water retention performance, can effectively adjust the proportion of water and gas and effectively convert nutrients becomes a hotspot.
The arbuscular mycorrhizal fungi are beneficial fungi in soil, can be symbiotic with 80% of terrestrial plant root systems to form arbuscular mycorrhiza, assist symbiotic plants to absorb water and mineral nutrition from the soil, promote plant growth and improve plant stress resistance. In agricultural production, most crops can be symbiotic with arbuscular mycorrhizal fungi to form arbuscular mycorrhiza, and the arbuscular mycorrhizal fungi has good effects on improving the crop yield and improving the crop variety. The method is beneficial to the formation of arbuscular mycorrhiza in the superior rhizosphere environment by inoculating arbuscular mycorrhizal fungi during seedling raising, and the crops carry the arbuscular mycorrhiza to enter a field during transplanting, so that the method has mature arbuscular mycorrhiza and stronger adaptability. However, the direct inoculation of the arbuscular mycorrhizal fungi into soil is easily affected by environmental factors and indigenous microorganisms to limit the action efficiency, and because the seedling culture substrate is a mixture of an organic substrate and an inorganic substrate, nutrients and water in the substrate can be utilized in the direct inoculation of the arbuscular mycorrhizal fungi, so that the strain activity is released in advance, and chemical substances in the organic fermentation product in the substrate can inhibit the arbuscular mycorrhizal fungi. Therefore, the existing arbuscular mycorrhizal fungi matrix has certain limitations in the aspects of mycorrhizal infection and activity preservation of mycorrhizal fungi. On the one hand, the infection rate is reduced because the propagules such as spores of arbuscular mycorrhizal fungi are too dispersed. On the other hand, arbuscular mycorrhizal fungi strains may germinate in advance in the shelf life and lose activity when not meeting the root system of the host plant.
Disclosure of Invention
The invention aims to provide a seedling culture medium containing arbuscular mycorrhizal fungi by adopting a smart carrier aiming at the problems in the prior art.
The invention also aims to provide a preparation method of the seedling raising substrate.
The invention also aims to provide application of the seedling raising substrate.
A seedling culture substrate containing arbuscular mycorrhizal fungi comprises an organic substrate, an inorganic substrate and encapsulated arbuscular mycorrhizal fungi strains, wherein the mass ratio of the arbuscular mycorrhizal fungi strains to the organic-inorganic substrate is (0.2-1): (99-99.8); the organic and inorganic matrix is formed by mixing compost fermentation products, perlite and vermiculite according to a volume ratio of (50-60) to (10-20) to (20-30).
As a preferred aspect of the present invention, said arbuscular mycorrhizal fungus is Rhizophora allowata (Rhizophagus irregularis); the arbuscular mycorrhizal fungi strain is prepared by the following method: the method comprises the steps of taking a mixture of sterilized soil and river sand in a mass ratio of 1: 1-3 as a substrate, symbiotically culturing two plant roots of heteroclitic radiculospora heteroclita and clover and corn for 100-105 days, harvesting, collecting the mixture of the substrate and the roots to obtain the arbuscular mycorrhizal fungi strain, wherein the effective components of the arbuscular mycorrhizal fungi strain comprise mature spores of the heteroclitic radiculospora and heteroclitic radiculospora hyphae existing in the substrate and the roots, and the density of the spores is more than or equal to 30 per gram of the substrate.
As a preferred mode of the invention, the compost fermentation product is prepared by the following method: straw, rice hull, wood dust, biogas residue, mushroom residue and wine residue are mixed according to the mass ratio of (30-40): (10-20): (5-15): (5-15): (5-15): (5-15) to obtain a substrate raw material, adding a fermentation microbial inoculum into the substrate raw material, wherein the fermentation microbial inoculum is a mixed strain of yeast, actinomycetes, lactic acid bacteria and bacillus, and the mass ratio of the fermentation substrate to the fermentation microbial inoculum is 480-550: 1, adjusting the water content to 60% -65%, and performing compost fermentation for 20-40 days to obtain a compost fermentation product.
As a further optimization of the invention, the mass ratio of the straw, the rice hull, the wood dust, the biogas residue, the mushroom residue and the wine residue in the fermentation substrate is 40: 20: 15: 15: 15: 15.
as a further preferred aspect of the present invention, the fermentation bacterial agent is prepared from a yeast bacterial agent, an actinomycete bacterial agent, a lactic acid bacterial agent and a bacillus bacterial agent according to a mass ratio of (28-30): (9-10): (8-12): (20-22) a mixed bacterial agent; the yeast is pichia pastoris, the actinomycetes is streptomyces albus, the lactic acid bacteria is lactobacillus acidophilus, and the bacillus is bacillus vallismortis.
Preferably, the mass ratio of the yeast agent, the actinomycete agent, the lactic acid bacteria agent and the bacillus agent in the fermentation agent is 30: 10: 10: 20.
preferably, the source and the preparation method of the fermentation inoculum are as follows: pichia pastoris, strain number SM168, available from Nippon island. The yeast is firstly cultured in a liquid culture medium, and the components of the culture medium are as follows: potato 20%, glucose 2%, natural pH value. When the culture reaches logarithmic growth phase, all the materials are transferred into a fermentation tank, and the fermentation medium comprises the following components: 3% of malt flour, 4% of glucose, 2% of sucrose and 0.3% of ammonium sulfate. The yeast in logarithmic phase is used as strain and inoculated into a fermentation tank, and the fermentation medium comprises 3% of soluble starch, 0.8% of soybean peptone, 0.05% of dipotassium hydrogen phosphate and 0.025% of magnesium sulfate.
Streptomyces albus, strain No. A31, was purchased from Nippon island Co. Scraping off a small amount of strain with an inoculating stick, and inoculating into liquid culture medium containing (g/L) soluble starch 20g, NaCl 0.5g, and KNO3 1g，K2HPO43H2O 0.5g，MgSO4 7H2O 0.5g，FeSO4·7H20.01g of O and 1000ml of water, and the pH value is 7.4-7.6. Placing on a constant temperature oscillator, and culturing at 28 deg.C and 240r/min for 24 h.
Lactobacillus acidophilus, strain No. NJ8, purchased from japan; the preparation method of the lactobacillus acidophilus microbial inoculum comprises the following steps: scraping a small amount of strains by using an inoculating rod, and inoculating the strains into a liquid culture medium, wherein the liquid culture medium comprises the following components in percentage by weight (g/L): glucose 20, peptone 10, yeast extract 5, beef extract 10 and KH2PO4 2,NaAc5,MnS04 0.2,MgSO40.2, 2 portions of ammonium citrate, and 6.2 to 6.6 portions of Tween-801 mL of pH value. Placing on a constant temperature oscillator, and culturing at 28 deg.C and 240r/min for 24 h. Inoculating Lactobacillus acidophilus in logarithmic phase as strain in a fermentation tank, wherein the fermentation medium comprises whey powder 12%, glucose 3%, and NH4H2P040.5 percent and the pH value is 6.2 to 6.6. And (6) detecting the pH value.
Bacillus vallismortis, strain No. MZ3, was purchased from Nippon island. The bacillus vallismortis is firstly cultured in a liquid culture medium, and the culture medium comprises the following components: peptone 1%, beef extract 1.5%, NaCl 1.5%, pH 7.2. When the culture is carried out to logarithmic growth phase, all the materials are inoculated into a fermentation tank, and the fermentation liquor comprises the following components: 1.3 percent of corn flour, 2.1 percent of bean cake powder, 0.7 percent of calcium carbonate and 0.03 percent of monopotassium phosphate.
As a further optimization of the invention, the volume ratio of the compost fermentation product, the perlite and the vermiculite in the organic matrix is 2:1: 1.
As a preferred aspect of the invention, each capsule encapsulates 1-2g of the arbuscular mycorrhizal fungal species.
The preparation method of the seedling substrate is characterized in that the arbuscular mycorrhizal fungi strain is filled into a capsule to obtain an arbuscular mycorrhizal fungi carrier, the capsule is added into the organic substrate to obtain the seedling substrate containing arbuscular mycorrhizal fungi, wherein the adding amount of the strain is 0.5-1%.
As a preferred aspect of the present invention, said arbuscular mycorrhizal fungal species is Rhizophora Heterophylla (Rhizophagus irregularis); the arbuscular mycorrhizal fungi strain is prepared by the following method: the method comprises the following steps of (1) taking a sterilized mixture of soil and river sand as a substrate, carrying out symbiotic culture on fungi and root systems of two plants, namely clovers and corns, culturing for 105 days, harvesting, and collecting the mixture of the substrate and the root systems to obtain the arbuscular mycorrhizal fungi strain, wherein the effective components of the arbuscular mycorrhizal fungi strain are mature spores and hyphae existing in the substrate and in the root systems, and the density of the spores is more than or equal to 30 per gram of the substrate; the compost fermentation product is prepared by the following method: straw, rice hull, wood dust, biogas residue, mushroom residue and wine residue are mixed according to the mass ratio of (30-40): (10-20): (5-15): (5-15): (5-15): (5-15) uniformly mixing to obtain a fermentation substrate, adding a fermentation microbial inoculum into the fermentation substrate, wherein the mass ratio of the fermentation substrate to the fermentation microbial inoculum is 500: 1, adjusting the water content to 60% -65%, and performing compost fermentation for 20-40 days to obtain a compost fermentation product.
The seedling culture substrate containing arbuscular mycorrhiza is applied to vegetable seedling culture or cultivation, and preferably applied to promotion of growth of watermelons and tomatoes.
In a preferred embodiment of the present invention, the capsule is a plant capsule, and the main raw material components thereof are lupulin, glycerin, purified water, and the like. The capsule consists of a cap and a body which are refined by the rupulan and auxiliary materials, and the specification size is 00. The capsule can be dissolved in water to release arbuscular mycorrhizal fungi. The water solubility of the capsules encapsulating the bacterial strain is shown in figure 2. After watering, the arbuscular mycorrhizal fungi are basically dissolved in 4 hours and completely dissolved in 8 hours, and then the arbuscular mycorrhizal fungi are released.
The present invention is applicable to all arbuscular mycorrhizal fungi and is not limited to Rhizopus irregularis.
The invention has the beneficial effects that:
1. according to the invention, the arbuscular mycorrhizal fungi is added into the organic matrix in a manner of encapsulating the arbuscular mycorrhizal fungi in the capsule, so that the direct contact between the arbuscular mycorrhizal fungi and the matrix is prevented, the inhibition effect of nutrients, especially phosphorus, in the matrix on the arbuscular mycorrhizal fungi is prevented, the moisture in the matrix is prevented from causing the arbuscular mycorrhizal fungi to germinate in advance in the shelf life of the matrix, and the strain activity and the inoculation potential are protected conveniently. Because the capsule has the characteristic of being dissolved in water, the capsule is quickly dissolved in the application process of the matrix, and arbuscular mycorrhizal fungi are released and germinate in time, so that the root system is conveniently infected, and an arbuscular mycorrhizal symbiotic structure is formed. Because the germination rate of the spores is increased along with the increase of the density of the spores in a certain range, the mycorrhizal fungi encapsulated by the capsules are intensively distributed after the capsules are dissolved, thereby promoting the germination of propagules such as arbuscular mycorrhizal fungi spores and the like and promoting the infection of crops.
2. The substrate contains arbuscular mycorrhizal fungi, so that the structure of the substrate is improved, the porosity and the water retention performance of the substrate are improved, the water-gas ratio is more effectively adjusted, and the nutrient effectiveness is further improved.
3. The substrate containing arbuscular mycorrhizal fungi is used for cultivating crops to form mycorrhizal seedlings, the seedlings of the crops cultivated by the substrate used by the method are stronger, the dry weight of the overground parts and the root systems of the seedlings is further improved, and the quality of melon seedlings is improved.
4. The raw materials are easy to obtain, the preparation method is simple, and the cost is low.
Drawings
FIG. 1 the substrate of the invention (right) has an extended shelf life, i.e.delayed germination of arbuscular mycorrhizal fungal spores. FIG. 1 shows that when the substrate with water content of 28% is left for 7 days under different inoculation modes, spores in the substrate germinate and hyphae are long. Left: when the arbuscular mycorrhizal fungi were directly mixed with the substrate, the spores were germinated with hypha length of 0.11mm after 7 days of storage (Table 2). The encapsulated arbuscular mycorrhizal fungi had no spores germinated and no hyphae grown (right).
Figure 2 the capsules in the matrix of the invention change over time at 100% moisture content. When the water content of the matrix of the capsule for encapsulating the arbuscular mycorrhizal fungi is 100%, the capsule starts to dissolve 2 hours after watering, dissolves 71.8% after 4 hours, and completely dissolves 8 hours after watering to release the arbuscular mycorrhizal fungi.
Figure 3 the capsules in the matrix of the invention change over time at different matrix water contents. The rate of dissolution of the capsules increases with increasing water content, and as can be seen from the data in table 1, the matrix dissolves completely within 5 days at 40% to 50% water content. The matrix is dissolved in water of 30% for 7-10 days. The matrix is dissolved after 10-20 days when the water content is 20%, and the matrix is dissolved after 20 days when the water content is 10%.
FIG. 4 shows the growth of tomato seedlings cultivated by the substrate of the present invention. The growth conditions of the tomato seedlings cultured by different matrixes are that under the culture condition of the encapsulated arbuscular mycorrhizal fungi matrixes, the tomato seedlings have robust root systems and overground parts, which are superior to seedlings cultured by directly mixing and inoculating the arbuscular mycorrhizal fungi matrixes and are superior to seedlings cultured by non-arbuscular mycorrhizal fungi matrixes.
FIG. 5 shows the growth of transplanted watermelon seedlings according to the present invention. The diagram A shows that the capsule arbuscular mycorrhizal fungi substrate promotes the growth of the watermelon, and the diagram B shows that the capsule arbuscular mycorrhizal fungi substrate improves the biomass of the upper part of the watermelon; FIG. C shows that the capsule arbuscular mycorrhizal fungi substrate improves the biomass of the watermelon root system; -AM, arbuscular mycorrhizal fungi substrate; + AM, common inoculation method arbuscular mycorrhizal fungi matrix; + JAM, capsule arbuscular mycorrhizal fungi matrix. The growing conditions of the transplanted watermelon seedlings cultured by the three matrixes are that the biomass of the overground part of the transplanted watermelon seedlings cultured by the matrixes is highest, and the growing conditions of the transplanted watermelon seedlings have a trend of being improved compared with watermelon plants cultured by directly mixed and inoculated arbuscular mycorrhizal fungi matrixes. The watermelon seedlings cultivated by the matrix have the highest biomass of the root system of the transplanted watermelon seedlings, and have an improved trend compared with watermelon plants cultivated by directly mixed inoculated arbuscular mycorrhizal fungi matrix.
Detailed Description
Example 1
1. And (3) preparing a matrix.
The straws are bought from farmer fields, the rice hulls, the saw dust, the biogas residues, the mushroom residues and the wine residues are bought from markets, the plant capsules are bought from markets, and the manufacturers are Shanxi Guangshi Capsule Limited companies.
(1) Uniformly mixing straw, rice hulls, wood chips, biogas residues, mushroom residues and wine residues, wherein the mass ratio of the straw, the rice hulls, the wood chips, the biogas residues, the mushroom residues and the wine residues is 40: 20: 15: 15: 15: 15, obtaining the matrix raw material.
(2) Adding a fermentation microbial inoculum into a substrate raw material, wherein the mass ratio of the fermentation substrate to the fermentation microbial inoculum is 500: 1, adjusting the water content to 60-65%, and fermenting the compost for 30 days to obtain a compost fermentation product. The fermentation inoculum is a mixed strain of yeast, actinomycetes, lactic acid bacteria and bacillus; the yeast is pichia pastoris, the actinomycetes is streptomyces albus, the lactic acid bacteria is lactobacillus acidophilus, the bacillus is bacillus vallismortis, and the mass ratio of the yeast to the actinomycetes to the lactic acid bacteria to the bacillus in the fermentation inoculant is 30: 10: 10: 20, in the composting fermentation process, if the temperature of the fermentation stack reaches 60 ℃, turning the stack. The fermentation product, i.e. the organic substrate, is obtained.
(3) Mixing the compost fermentation product with perlite and vermiculite, wherein the compost fermentation product, the perlite and the vermiculite are mixed according to the volume ratio of 60: 10: 30, and mixing uniformly to obtain the organic-inorganic matrix raw material.
(4) The substrate material was autoclaved (121 ℃, 1h) to kill the microorganisms in the substrate.
(5) The arbuscular mycorrhizal fungi were encapsulated in capsules, each capsule carrying 1g of arbuscular mycorrhizal fungi strain.
(6) And (3) mixing the matrix obtained by the above steps: the mass ratio of the arbuscular mycorrhizal fungi is 100: 1, mixing to obtain the substrate containing the arbuscular mycorrhizal fungi.
2. The capsule dissolution rate can be regulated: the dissolution rate increases with increasing water content
The treatment method comprises the following steps: the arbuscular mycorrhizal fungi are encapsulated in capsules, each capsule containing 1g of arbuscular mycorrhizal fungi strain. Mixing a substrate and an arbuscular mycorrhizal fungi strain according to the mass ratio of 100: 1, mixing to obtain the substrate containing the arbuscular mycorrhizal fungi. The substrate water content was set at 10%, 20%, 30%, 40%, 50%, ten times per treatment, placed in a constant temperature incubator for dark culture at 25 ℃, and the change in area of the support encapsulating arbuscular mycorrhizal fungi was measured at 0d, 2d, 3d, 4d, 5d, 6d, 10d, 20d, respectively, of the culture. Two treatments, one for direct inoculation of non-encapsulated capsules with arbuscular mycorrhizal fungi and one for encapsulation of arbuscular mycorrhizal fungi, were set, each treatment being repeated ten times.
Capsule dissolution characteristics: as shown in Table 1, the substrates of the present invention, carriers encapsulating arbuscular mycorrhizal fungi, were subjected to a melting time at various water contents of the substrates. When the water content of the substrate is 40% -50%, the carrier can be stored for 4 days; the water content of the substrate is below 30%, and the mycorrhizal fungi carrier can be preserved for more than 6 days. Therefore, in a natural state, the matrix containing the carrier for encapsulating the arbuscular mycorrhizal fungi can be directly mixed at present as required, and can also be mixed in advance according to the physicochemical properties of the matrix.
TABLE 1 Carriers encapsulating arbuscular mycorrhizal fungi in the Natural State vary in area (area: cm) under different water content conditions in the substrate2)
3. The capsule has prolonged germination time of mycorrhizal fungi and prolonged shelf life.
The treatment method comprises the following steps: the substrate containing the arbuscular mycorrhiza is placed in a constant-temperature incubator for dark culture at 25 ℃, and the spore germination rate and the hypha length are detected on the 7 th day, the 14 th day and the 21 th day of the culture. The results are shown in table 2, and the substrate of the invention has a significant lag in spore germination rate and hyphal length compared with the substrate in which non-encapsulated capsules are directly inoculated with arbuscular mycorrhiza. At 7 days of treatment, arbuscular mycorrhizal spores in the conventionally mixed mycorrhizal substrate had germinated and hyphae began to grow, while spores of the substrate of the present invention did not germinate and hyphae did not grow. The spore germination rate and hypha length of the substrate of the invention are significantly lower than those of the directly mixed arbuscular mycorrhizal substrate at 14 days of treatment. Compared with the directly mixed arbuscular mycorrhizal fungi matrix 21 days after treatment, the matrix of the invention has consistent spore germination rate, and the hypha length is obviously shorter than that of the mycorrhizal fungi matrix produced by the direct mixing mode.
The quality guarantee period of the matrix is characterized in that: the spore germination is delayed for more than 7 days, and the time is equivalent to 21 days in a direct mixing mode. The hyphal length was still lower than that of the directly mixed matrix by 21 days. Therefore, in a natural state, the encapsulation of the arbuscular mycorrhizal fungi matrix is more beneficial to the preservation of the activity of the arbuscular mycorrhizal fungi in the transportation and preservation processes.
TABLE 2 Change in spore germination rate and hypha length of arbuscular mycorrhizal fungi in Encapsulated capsules in Natural State
Example 2: strong seedling cultivated by arbuscular mycorrhizal seedling raising medium
1. Test method
Soaking seeds of watermelon and tomato, sterilizing, accelerating germination, and after the seeds germinate and emerge white, selecting full and uniform seeds to be sown in 50-hole trays filled with mixed substrates, wherein one seed is in each hole. After the seeding is finished, the hole tray is placed in a glass greenhouse. During seedling raising, according to the moist state of the seedling raising matrix and the growth state of seedlings, the water content of the matrix is irregularly supplemented by tap water, and the watering amount of each plug tray is based on that the watered water does not leak out of the plug tray.
2. Test results
Table 3 shows that compared with a substrate containing no arbuscular mycorrhizal fungi, the arbuscular mycorrhizal seedling substrate provided by the invention has the advantages that the plant height, stem thickness, overground part dry weight, underground part dry weight, crown-root ratio, leaf area and mycorrhizal infection rate of tomato seedlings are remarkably improved. Compared with the matrix which is directly inoculated with arbuscular mycorrhizal fungi without capsule encapsulation, the mycorrhizal infection rate is equivalent, the plant height and the stem thickness are obviously improved; the dry weight of the above ground part and the dry weight of the below ground part are increased, but the difference is not obvious. FIG. 4 shows that tomato seedlings under the conditions of the medium culture of arbuscular mycorrhizal fungi in capsules are the most robust. Therefore, the arbuscular mycorrhizal seedling substrate has advantages in cultivating tomato seedlings.
TABLE 3 Effect of the substrate of the present invention for raising tomato seedlings
Table 4 shows that the arbuscular mycorrhizal seedling substrate provided by the invention is used for culturing watermelon seedlings. After the watermelon seedlings are cultured for 28 days, compared with a matrix containing no arbuscular mycorrhizal fungi, the watermelon seedlings cultured by the matrix are remarkably improved in plant height, stem thickness, dry weight of the overground part, dry weight of the underground part, crown root ratio, leaf area, total root length and mycorrhizal infection rate. Compared with the arbuscular mycorrhizal fungi matrix inoculated in a common mixing mode, the capsule arbuscular mycorrhizal fungi matrix has the advantages that the mycorrhizal infection rate is equivalent, and the plant height is obviously improved.
TABLE 4 Effect of the substrate of the present invention for cultivation of watermelon seedlings
Example 3: the advantages of the watermelon seedlings cultivated by the arbuscular mycorrhizal seedling medium after transplantation
The watermelon seedlings cultured by the seedling culture medium prepared in the example 1 have better growth conditions than the watermelon seedlings cultured by the common seedling culture medium 30 days after transplantation (figure 5A), the watermelon seedlings cultured for 28 days are transplanted into potted plants potted by the mixed medium, and three watermelon seedlings are transplanted in each pot. According to the wet state of the matrix and the growth state of the watermelon seedlings, the water content of the matrix is supplemented by tap water irregularly, and the watering amount of each pot is based on that the watered water does not leak out of the pot. The effect of the substrate containing the arbuscular mycorrhizal fungi is verified by measuring the growth indexes of the watermelons after one month of transplantation. Fig. 5B and fig. 5C show that the dry weight of the upper part and the dry weight of the lower part of the watermelon cultivated by the arbuscular mycorrhizal seedling substrate of the invention are significantly higher than those of the watermelon cultivated by the common seedling substrate, and compared with the watermelon cultivated by the substrate directly connected with the arbuscular mycorrhizal, the dry weight of the upper part and the dry weight of the lower part of the watermelon cultivated by the arbuscular mycorrhizal seedling substrate of the invention are increased. Therefore, the arbuscular mycorrhizal seedling substrate has advantages in cultivating watermelon seedlings.
The above-mentioned experiments are only for illustrating the concept and characteristics of the present invention, and the main purpose is to disclose a substrate containing arbuscular mycorrhizal fungi, which is known and practiced by those skilled in the art, and should not be construed as limiting the scope of the present invention, and all modifications or variations made according to the spirit of the present invention should be covered by the scope of the present invention.
Claims (13)
1. A seedling substrate containing arbuscular mycorrhizal fungi is characterized in that the seedling substrate containing the arbuscular mycorrhizal fungi consists of an organic-inorganic substrate and encapsulated arbuscular mycorrhizal fungi strains, and the mass ratio of the arbuscular mycorrhizal fungi strains to the organic-inorganic substrate is (0.2-1): (99-99.8); the organic substrate is a compost fermentation product, and the inorganic substrate is perlite and vermiculite; the compost fermentation product is as follows: perlite: the volume ratio of the vermiculite is (40-60): (10-20): 20-30); the seedling raising substrate is prepared by the following method: encapsulating the arbuscular mycorrhizal fungi strain to obtain an arbuscular mycorrhizal fungi carrier, and adding the capsule into the organic-inorganic matrix to obtain a seedling culture matrix containing arbuscular mycorrhiza; the capsule consists of a cap and a body two-section capsule shell which are refined by adding auxiliary materials into the rupulan, the rupulan is basically dissolved after being watered for 4 hours, and the rupulan is completely dissolved after 8 hours to release arbuscular mycorrhizal fungi; the method comprises the steps of adding the arbuscular mycorrhizal fungi into an organic matrix in a manner of encapsulating the arbuscular mycorrhizal fungi in a capsule, preventing the arbuscular mycorrhizal fungi from being in direct contact with the matrix, preventing nutrient phosphorus in the matrix from inhibiting the arbuscular mycorrhizal fungi, preventing moisture in the matrix from causing the arbuscular mycorrhizal fungi to germinate in advance in the shelf life of the matrix, and facilitating protection of strain activity and inoculation potential.
2. A growth substrate according to claim 1, characterised in that the arbuscular mycorrhizal fungus is rhizospora heterosporum (Rhizophagus irregularis); the arbuscular mycorrhizal fungi strain is prepared by the following method: the method comprises the steps of taking a mixture of sterilized soil and river sand in a mass ratio of 1: 1-3 as a substrate, symbiotically culturing two plant roots of heteroclitic radiculospora heteroclita and clover and corn for 100-105 days, harvesting, collecting the mixture of the substrate and the roots to obtain the arbuscular mycorrhizal fungi strain, wherein the effective components of the arbuscular mycorrhizal fungi strain comprise mature spores of the heteroclitic radiculospora and heteroclitic radiculospora hyphae existing in the substrate and the roots, and the density of the spores is more than or equal to 30 per gram of the substrate.
3. A growth substrate according to claim 1, characterized in that said compost fermentation product is prepared by the following method: uniformly mixing straw, rice hulls, sawdust, biogas residues, mushroom residues and wine residues to obtain a fermentation substrate, adding a fermentation microbial inoculum into the fermentation substrate, wherein the fermentation microbial inoculum is a mixed microbial inoculum of saccharomycetes, actinomycetes, lactic acid bacteria and bacillus, and the mass ratio of the fermentation substrate to the fermentation microbial inoculum is 480-550: 1, adjusting the water content of the fermentation substrate to be 60% -65%, and performing compost fermentation for 20-40 days to obtain a compost fermentation product.
4. A seedling raising substrate according to claim 3, wherein the fermentation substrate comprises straw, rice hulls, sawdust, biogas residues, mushroom residues and wine residues in a mass ratio of (30-40): (10-20): (5-15): (5-15): (5-15): (5-15).
5. A seedling substrate according to claim 3, wherein the fermentation inoculant is prepared from a yeast inoculant, an actinomycete inoculant, a lactic acid inoculant and a bacillus inoculant in a mass ratio of (28-30): (9-10): (8-12): (20-22) mixing; the yeast is pichia pastoris, the actinomycetes is streptomyces albus, the lactic acid bacteria is lactobacillus acidophilus, and the bacillus is bacillus vallismortis.
6. A seedling substrate according to any one of claims 1-5, characterized in that the volume ratio of the compost fermentation product, perlite and vermiculite in the organic substrate is 2:1: 1.
7. A growth substrate according to claim 1 or 2, characterized in that each capsule encapsulates 1-2g of arbuscular mycorrhizal fungal species.
8. A method for preparing a seedling substrate according to claim 1, characterized in that the arbuscular mycorrhizal fungi strain is encapsulated to obtain an arbuscular mycorrhizal fungi carrier, and the capsule is added into the organic-inorganic substrate to obtain a seedling substrate containing arbuscular mycorrhizal fungi, wherein the addition amount of the strain is 0.2-1%; the organic and inorganic matrix is formed by mixing the organic matrix and the inorganic matrix in proportion.
9. The method according to claim 8, wherein the arbuscular mycorrhizal fungal species is Rhizoctonia heterodera; the arbuscular mycorrhizal fungi strain is prepared by the following method: the method comprises the following steps of (1) taking a mixture of sterilized soil and river sand in a mass ratio of 1: 1-3 as a substrate, symbiotically culturing fungi with roots of two plants, namely clovers and corns, culturing for 100-105 days, harvesting, and collecting a mixture of the substrate and the roots to obtain the arbuscular mycorrhizal fungi strain, wherein the effective components of the arbuscular mycorrhizal fungi strain are mature spores and hyphae existing in the substrate and in the root system, and the density of the spores is more than or equal to 30 per gram of the substrate; the compost fermentation product is prepared by the following method: straw, rice hull, wood dust, biogas residue, mushroom residue and wine residue are mixed according to the mass ratio of (30-40): (10-20): (5-15): (5-15): (5-15): (5-15) uniformly mixing to obtain a fermentation substrate, adding a fermentation microbial inoculum into the fermentation substrate, wherein the mass ratio of the fermentation substrate to the fermentation microbial inoculum is 500: 1, adjusting the water content to 60% -65%, and performing compost fermentation for 20-40 days to obtain a compost fermentation product.
10. Use of a seedling raising substrate containing arbuscular mycorrhiza according to claim 1 in vegetable cultivation.
11. Use according to claim 10, characterized in that the use of a growth substrate comprising arbuscular mycorrhiza for promoting the growth of tomatoes.
12. The application of claim 11, wherein a suitable seedling raising tray is selected, the seedling raising substrate containing the arbuscular mycorrhizal fungi is filled, seeds with germination accelerating and whitening effects are sown, the water content is 100%, the capsule is completely dissolved within 7-9 hours, the arbuscular mycorrhizal fungi are released, the arbuscular mycorrhizal fungi and the seeds grow simultaneously, the arbuscular mycorrhizal fungi are formed through recognition and infection, the water content in the substrate is kept between 15% and 80% through normal water irrigation management, and strong seedlings are raised.
13. Use of a seedling raising substrate containing arbuscular mycorrhiza according to claim 1 for the growth of watermelons.
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| CN114747437B (en) * | 2022-04-12 | 2023-08-25 | 浙江省农业科学院 | Method for relieving toxic action of cadmium in tomatoes by using arbuscular mycorrhizal fungi and application |
| CN114804961A (en) * | 2022-06-10 | 2022-07-29 | 上海丰野实业有限公司 | Bacterial fertilizer, intraradicular sporangium capsule preparation containing bacterial fertilizer, and preparation method and application of bacterial fertilizer |
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