CN114504001A - Seed coating agent of masson pine ectomycorrhizal fungi, preparation and use method thereof - Google Patents
Seed coating agent of masson pine ectomycorrhizal fungi, preparation and use method thereof Download PDFInfo
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
- A01C1/06—Coating or dressing seed
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
The invention discloses a masson pine ectomycorrhizal fungi seed coating agent, which comprises the following components: 5-30 parts of a water-retaining agent of the ectobacteria liquid; 5-20 parts of a bonding agent and 4-8 parts of humus soil; 20-50 parts of growth regulator 0.005-0.03 part; 30-55 parts of water. A preparation method of a seed coating agent of masson pine ectomycorrhizal fungi; mixing mycelium liquid containing Boletus viscosus, Lasiosphaera leucocephala and Lactarius deliciosus, humus, gibberellin and viscosity agent, stirring, slowly adding purified water, stirring, stopping adding water, and standing to obtain a solidified state. The using method comprises the following steps: seed disinfection: preparing 2L of 600 times of wettable powder of 75 percent chlorothalonil with good adhesion, broad-spectrum sterilization and low toxicity, soaking the seeds for 1h, and washing the seeds for later use by distilled water for 3 times; seed coating: and putting the disinfected seeds and the mixed seed coating agent into a seed coating machine for coating, wherein the mass ratio of the seed coating agent to the seeds is 2: 1. The method has the characteristics of promoting the germination rate of seeds, promoting the growth of seedlings and improving the resistance of seedlings.
Description
Technical Field
The invention relates to the field of agriculture and forestry, in particular to a masson pine ectomycorrhizal fungi seed coating agent, a preparation method and a use method thereof.
Background
The seed coating agent is prepared by mixing mycelium of ectomycorrhizal fungi, fertilizer, growth regulator, humus soil and other active substances in certain proportion, coating the seed with adhesive and other inactive substances, and curing to form coating film with certain permeability. The seed coating agent for the ectomycorrhizal fungi not only has the advantages of long preservation time, slow release of fertilizer effect, good protection effect, environmental protection, safety and the like, but also can promote seed germination and root system group formation, improve seedling resistance, prevent diseases and insect damage and the like, and finally form mycorrhizal seedlings. Seed coating research begins in the 60's of the 20 th century, and the development and application of conventional seed coatings have been limited to crop seeds for a long time, while forest seeds are relatively lacked in related exploration due to various tree types and large differences in biological characteristics. In general, no reports have been made on the seed coating agent of ectomycorrhizal fungi of forest trees.
The masson pine is an important fast-growing material and a resin-collecting resin species, and has higher economic value. The masson pine not only has the widest distribution area and the largest quantity in the south of China, but also has the advantages of high yield, drought resistance, soil barren resistance, strong adaptability and the like, and is also the preferred tree species in barren mountain greening and vegetation construction. The masson pine is mainly used for sampling seeds to propagate and grow seedlings. In the process of seedling cultivation, seeds are easily damaged by diseases, pests, wild animals and the like, and the seedling growth potential is weak or the survival rate is low due to insufficient moisture and nutrients after seedling formation and in the growth process. Therefore, the seed germination, the seedling development condition in the seedling development stage and the rhizosphere microenvironment are key factors for the success and failure of the Pinus massoniana seedling. After the pinus massoniana seeds germinate, root tissues are tender and small in quantity, the functions of absorbing water and nutrients are still not perfect, and the death rate of seedlings is high in the processes of drought damage, waterlogging, disease and insect damage and the like. Research has shown that after pinus massoniana seedlings are inoculated with exogenous mycorrhiza, the mycorrhiza seedlings are obviously improved in aspects of seedling height, ground diameter, lateral root number, dry weight, resistance and the like (Sun folk music, etc. the influence of exogenous mycorrhiza fungi on the emergence and growth of different pine trees, Nanjing university of forestry, proceedings of Nature science, 2007(05): 39-43). However, since this method is limited to the potted propagation of the masson pine mycorrhizal seedlings, the roots of the seedlings are still damaged and lost in the later period by transplanting the seedlings to the forestation area, and a seedling revival period for the growth of the seedlings is formed, resulting in slow growth in the seedling period. This is also a drawback that this technique cannot avoid.
The patent of the invention with application number of CN202010276810.5 discloses that the ectomycorrhiza is stuck on the root system of host seedling and then placed in greenhouse or incubator to be cultured into mycorrhiza seedling. Although the cultivation of mycorrhizal seedlings can be completed by applying the method, certain professional equipment is required to be utilized in the seedling cultivation process, so that the operation process is complex and the cost is high. Meanwhile, the invention patent No. CN110754483A discloses that the drought-resistant water-retaining agent, the adhesive, the soil conditioner, the humus and the growth regulator are uniformly mixed and stirred to be sticky and used as a masson pine seed coating agent, the germination rate of seeds and the drought resistance of seedlings can be improved, but the application is only limited to cultivation of general seedlings. Because the composition of the strain lacks of strain microorganisms necessary for culturing mycorrhizal seedlings, the strain cannot form higher-quality mycorrhizal seedlings. Therefore, the seed coating agent which is simple and convenient to operate and low in cost and can be used for cultivating the masson pine mycorrhizal seedlings is very necessary.
Disclosure of Invention
Aiming at the defects of the technology, the invention aims to develop the coating agent for cultivating the ectogenic strains of the pinus massoniana seeds, and the coating agent has the characteristics of improving the germination rate of the seeds, promoting the growth of seedlings and improving the resistance of seedlings.
The first aim is realized, and the invention provides the following technical scheme:
the invention provides a masson pine ectomycorrhizal fungi seed coating agent which comprises the following raw materials in parts by weight:
5-30 parts of ectotrophic bacterium liquid, 5-20 parts of water-retaining agent, 4-8 parts of adhesive, 20-50 parts of humus, 0.005-0.03 part of growth regulator (gibberellin) and 30-55 parts of water.
Boletus viscosus; colorful bean puffball; d, lactarius deliciosus; three strains are collected in soil under the artificial forest of 35 years masson pine in Longli forest farm in China in Longli forest of Longli county of Guizhou province in 2020, 25 days, the separated strains are stored in a forest resource and environment research center of Guizhou university (called Guizhou university forest center for short, address: Chongde building of Huaxi district of Guiyang city of Guizhou province, A city, south China university, West school district of Guizhou university; storage date: 2020, 7 months and 25 days) microbial resource laboratory, and the serial numbers are M20200725005, M20200725007 and M20200725008 respectively.
See example 1 for an illustration, now stored in the Guizhou province forest resources and environmental research center laboratory at Guizhou university.
By the technical scheme, 3 mycelium bacteria liquids of the ectomycorrhizal fungi are added into the raw materials for preparing the seed coating agent, which is a necessary condition for forming mycorrhizal seedlings. The Boletaceae Boletus genus (Suillus); the diameter of the pileus is 3-10cm, the pileus is hemispherical, and the edge of the pileus is thinned; smooth surface, earthy yellow or light yellow brown; it is sticky when wet and lustrous after drying out. The colorful bean puffball (Pisolithus tinctorius) is an ectomycorrhizal fungus of Scleroderma puffball family (Sclerodermaceae) genus (Scleroderma), the fruiting body of the colorful bean puffball is spherical to oblate spherical, the width is 2-10 cm, the base part is reduced to handle shape, the yellow brown color is changed to dark brown color, and the coating film is smooth. Lactarius deliciosus (Lactarius) is a Lactarius (Lactarius) fungus of Russulaceae (Russulaceae), the pileus is hemispherical when young, the middle is concave, the pileus is flat and concave when mature, the diameter is 3-12 cm, and the middle part is 8-20 mm thick; the mycelial hyphae are colorless, thin-walled to slightly thick-walled, and have few branches. The 3 ectomycorrhizal fungi fruiting bodies are mainly grown on pines or other coniferous forests in summer and autumn and are clustered or gregarious; the mycelium of the 3 plants can form a mycorrhizal complex with the root of the higher plant, so that the number of lateral roots is effectively increased, the surface area of the root system is increased, and the experimental result is shown in figure 1. Meanwhile, metabolites of host plants and mycorrhizal fungi are bidirectionally operated through a Hardgrove net, so that the capacities of absorbing and transporting water and nutrients of plant root systems can be obviously enhanced, the plant growth is promoted, and the abiotic stress resistance of seedlings is improved.
The humus contains a large amount of macromolecular organic compounds, can obviously improve the soil structure, and increase and maintain the soil labor. The addition of humus can improve more sufficient nutrient species for the germination of the pinus massoniana seeds, and the loose structure provides oxygen required by respiration during the seed germination process, so that the seed germination is promoted. The growth regulator is favorable for accelerating seed metabolism, cell division and differentiation and improving germination potential and germination rate.
The growth regulator is gibberellin, which functions to break seed dormancy and promote its germination. Meanwhile, gibberellin can promote plant cell elongation and differentiation and accelerate plant growth.
The seed coating agent of the invention is provided with the water-retaining agent in the raw material, which can absorb and store the moisture in the environment, and can be slowly released when the environment is dry and lacks water, so as to solve the problems of pinus massoniana seed germination and moisture shortage in the seedling growth process. The adhesive is water-soluble polyvinyl alcohol, can increase the viscosity of the mixture of various substances, but has no influence on the functions and the characteristics of the substances.
The water-retaining agent is polyvinyl alcohol and water-absorbent resin SAP, wherein the water-absorbent resin SAP is generally high in price, and in addition, if the polyacrylamide water-absorbent resin is degraded after a certain time, the coagulation effect is not long enough; the polyvinyl alcohol has good and long-term coagulability, is low in price and easy to obtain, so that the water retaining agent is preferably polyvinyl alcohol.
Through the selection of the raw materials, the hypha liquid of the bolete multocida, the colorized bean puffball and the lactarius deliciosus and the humus are respectively used as the raw materials, mixed with gibberellin and water-retaining agent polyvinyl alcohol, added with a viscosity agent and dissolved into a certain amount of purified water together to form a colloid paste to wrap the seed of the pinus massoniana. The mycelium, organic matters, mineral matters and water around the seeds are fixed, so that the vitality maintenance and germination requirements of the masson pine seeds are met. Gibberellin can break seed dormancy, and can promote cell elongation and plant growth after seeds germinate.
The second purpose of the invention is to provide a preparation method of the masson pine ectomycorrhizal fungi seed coating agent, which has the advantages of simple operation, low cost and convenient and efficient preparation of the masson pine seed coating agent.
In order to achieve the second object, the invention provides the following technical scheme:
the invention provides a preparation method of a masson pine ectomycorrhizal fungi seed coating agent, which comprises the following steps:
1) the preparation process of the boletus mucosae, the columba esculenta and the lactarius deliciosus comprises the following steps: respectively filling the three bacteria into a sterilized triangular flask with 500ML culture solution, and then pricking the bottle openings with 6 layers of sterile gauze; performing shaking culture at 25 deg.C and 120 times/min amplitude for 7 days to obtain 3 kinds of dense mixed fungi mycelia;
2) mixing mycelium liquid containing Boletus viscosus, Lasiosphaera leucocephala and Lactarius deliciosus, humus, gibberellin and viscosity agent, stirring, slowly adding purified water, stirring, stopping adding water, and standing to obtain a solidified state.
By adopting the technical scheme, after the preparation process is regulated and controlled to obtain the sticky mixture, the sticky mixture is placed for 20-30min until the sticky mixture is solidified, the operation process is simple and easy to implement, and the prepared seed coating agent has an obvious pore structure, so that the seed coating agent has the water and fertilizer retention performance. The seed germination rate, drought resistance and root development effect of the masson pine coated by the seed coating agent are better. The preparation process adds the strain innovatively, which is a necessary condition for forming mycorrhizal seedlings; secondly, the long-term survival of the strain requires certain moisture conditions. The bacteria coating agent still keeps certain humidity after solidification, namely contains a certain amount of water, so that the strains can keep better life.
The third purpose of the invention is to provide a using method of the masson pine ectotrophic mycorrhizal seed coating agent, which has the condition of germinating and forming mycorrhizal seedlings after the masson pine seeds are coated with the ectotrophic mycorrhizal coating agent, and has the advantages of simple, efficient and low-cost cultivation of the masson pine ectotrophic mycorrhizal seedlings.
In order to achieve the third objective, the invention provides the following technical solutions:
the invention provides a using method of a masson pine ectomycorrhizal fungi seed coating agent, which comprises the following steps:
1) seed disinfection: preparing 2L of 600 times of wettable powder of 75 percent chlorothalonil with good adhesion, broad-spectrum sterilization and low toxicity, soaking the seeds for 1h, and washing the seeds for later use by distilled water for 3 times;
2) seed coating preparation: mixing mycelium liquid containing Boletus viscosus, Lasiosphaera leucocephala and Lactarius deliciosus, humus, gibberellin and mucoadhesive, stirring, slowly adding purified water, stirring to obtain paste, stopping adding water, and standing to obtain a solidified state;
3) seed coating: placing the seeds for standby disinfection and the mixed seed coating agent into a seed coating machine for coating, wherein the mass ratio of the seed coating agent to the sterile seeds is (0.2-2): 1.
By adopting the scheme, when the mass ratio of the seed coating agent to the seeds is 2:1, the germination rate, the drought resistance and the lateral root number of the masson pine seeds are higher.
In summary, the invention has the following advantages:
firstly, the invention adopts the hypha liquid of 3 kinds of ectomycorrhizal fungi as the source of the ectomycorrhizal fungi, the pinus massoniana forest is naturally distributed, the prepared bacterial liquid has strong fluidity and permeability, and the invention has the advantages of convenient acquisition of raw materials, low cost, high root system impregnation rate and the like, thereby leading the success rate of the cultivation of mycorrhizal seedlings to be higher.
Secondly, humus soil, a water-retaining agent, gibberellin and the like are adopted as preparation raw materials of the masson pine seed coating agent, different materials are matched with one another, and the prepared seed coating agent is good in viscosity, strong in water and fertilizer holding capacity, capable of providing excellent conditions for seed germination and seedling growth, non-toxic, environment-friendly and outstanding in advantages.
Thirdly, the coating method has simple and easy operation steps, and the ectomycorrhizal fungi seed coating agent is applied to the masson pine seeds, so that the coated seeds have stronger vitality, water retention and abiotic stress resistance, and are easier to germinate and form mycorrhizal seedlings.
Drawings
FIG. 1 is a statistical chart of germination of pinus massoniana seeds coated with ectomycorrhizal fungi of examples 1-6;
FIG. 2 is a statistical chart of the germination of the pinus massoniana seeds coated with the ectomycorrhizal fungi of comparative examples 1-5.
Detailed Description
The preparation and the application of the masson pine ectomycorrhizal fungi seed coating agent provided by the invention are further explained in more detail through specific embodiments as follows:
example 1
The masson pine seed coating agent of the embodiment has the following formula:
boletus viscosus, Cobalata colorata and Lactarius deliciosus hyphae solutions were mixed according to a ratio of 1: 1: 1 volume ratio, preparing 10Kg of mixed mycelium liquid, 5Kg of water-retaining agent, 5Kg of binder, 20Kg of humus, 0.005Kg of growth regulator and 50Kg of water.
Wherein the mycelium liquid of Boletus viscosus, Lasiosphaera leucotricha and Lactaria Pinus is mycelium of mother strain collected from Larix Gmelini forest in Longli county of Qian south of Guizhou province. The bottles were separately filled with 500ML of culture medium and then sealed with 6 layers of sterile gauze. Culturing at 25 deg.C with oscillation amplitude of 8cm and frequency of 120 times/min for 7 days to grow dense mycelium of 3 kinds of mixed fungi in liquid.
The water-retaining agent is purchased from Ningchu Cihuida chemical Co., Ltd, and has the model of HD2411 and the particle size of 100 meshes.
The adhesive polyvinyl alcohol is purchased from treasure building materials science and technology limited of Jinzhou city, Hebei province, and has the model of PVA2488 and the grain size of 160 meshes.
The humus soil is derived from the humus layer of masson pine forest in Longli county of Guizhou province.
The application method of the masson pine ectomycorrhizal fungi seed coating agent comprises the following steps:
seed disinfection: soaking and disinfecting masson pine seeds by using 600 times of low-toxicity broad-spectrum 75% chlorothalonil wettable powder for 1 hour, and then washing the masson pine seeds by using distilled water for 3 times for later use;
seed coating preparation: mixing water-retaining agent, adhesive, humus soil and growth regulator, stirring, adding purified water, stirring to obtain paste, standing for 25min to obtain gel;
seed coating: and (3) putting the sterile seeds and the seed coating agent into a coating machine according to the mass ratio of 5:1, starting the coating machine to respectively rotate clockwise and counterclockwise for 2min, and then continuously operating the coating agent to respectively rotate clockwise and counterclockwise for 5 times to finish seed coating.
First, embodiments 2 to 6 are different from embodiment 1 in that: the seed coating agent of the masson pine ectomycorrhizal fungi of the embodiments 2-6 has different preparation formulas and proportions, and the proportion scheme is shown in table 1.
TABLE 1 example 1-6 recipe ratio (Kg) of seed coating agent for masson pine ectomycorrhizal fungi
| Item | Ectomycorrhizal fungi | Water-retaining agent | Adhesive agent | Humus soil | Gibberellins | Purified water |
| Example 1 | 10 | 5 | 5 | 20 | 0.005 | 50 |
| Example 2 | 20 | 10 | 8 | 40 | 0.02 | 40 |
| Example 3 | 5 | 15 | 5 | 30 | 0.02 | 55 |
| Example 4 | 15 | 20 | 5 | 30 | 0.01 | 45 |
| Example 5 | 25 | 10 | 4 | 50 | 0.03 | 35 |
| Example 6 | 30 | 15 | 4 | 40 | 0.02 | 30 |
The specific operation and implementation steps of the preparation method and the coating method of the masson pine ectomycorrhizal fungi seed coating agent of the embodiment 2-6 are the same as those of the embodiment 1.
Comparative examples 1-5 differ from example 2 in the formulation and formulation ratio for preparing the masson pine ectomycorrhizal fungi seed coating agent, as shown in table 2, and in the preparation, coating operation and procedure identical to example 2.
TABLE 2 COMPARATIVE EXAMPLE 1-5 FORMULATION (Kg) OF SEEDLING OF MALEISONG EXTERNAL-RUBBING BACTERIA COULDING AGENT
| Item | Ectomycorrhizal fungi | Water-retaining agent | Adhesive agent | Humus soil | Gibberellins | Purified water |
| Example 2 | 20 | 10 | 8 | 40 | 0.02 | 40 |
| Comparative example 1 | 0 | 10 | 8 | 40 | 0.02 | 40 |
| |
20 | 0 | 8 | 40 | 0.02 | 40 |
| Comparative example 3 | 20 | 10 | 0 | 40 | 0.02 | 40 |
| Comparative example 4 | 20 | 10 | 8 | 0 | 0.02 | 40 |
| Comparative example 5 | 20 | 10 | 8 | 40 | 0 | 40 |
Second, test of seed germination and seedling development
The masson pine seeds are coated by proportioning the formulas of examples 1-6 and comparative examples 1-5 and preparing a seed coating agent, and 100 masson pine seeds are selected for coating in each example and comparative example. Each recipe is considered a process, with 3 replicates per process set. The wrapped masson pine seeds were sown at equal intervals in pots (specification: diameter 25 cm. times.height 15cm) with 10cm thick sandy soil, and the seeds were slightly pressed into the sandy soil to facilitate observation of the germination of the seeds.
And recording the germination number of the seeds every day from the 5 th day of the experiment, continuously observing until the 50 th day, and finishing when the germination number is continuously unchanged for 3 days, measuring the height of the seedlings by using a steel tape measure during the last observation, taking out the seedlings, and washing the roots by using clear water to count the number of lateral roots. And (3) observing the infection condition of the root system of the seedling under a microscope, wherein if the seedling is infected by the ectomycorrhizal fungi, the seedling is a mycorrhizal seedling, and otherwise, the seedling is a common seedling.
And calculating the germination rate, the germination vigor, the germination index and the vitality index of the pinus massoniana seeds, and counting the height and the lateral root number of the seedlings.
The germination rate (%) of the seeds is s/Nx 100%;
the seed germination potential (%) ═ Gf/N × 100%;
germination index (Zi) ═ Sigma (Z) of seedsi/Dt)×100％；
Seed vigor index (V) ═ L/Zi
Seedling average height (H) ═ Sigma Hi/N；
Side number (R) ═ Σ Ri/N；
The ratio (%) of ectomycorrhizal fungi to young root ═ Σ Ji/N
Wherein s is the number of normal germination seeds, and N is the total number of seeds to be tested; gf is the total number of sprouts within t days; dt is germination days, ZiIn order to achieve the total germination number of the seeds in the germination peak period, L is the height of the seedlings, Hi is the height of the seedlings after germination, and Ri is the number of lateral roots of the seedlings after germination; j. the design is a squareiThe number of the ectomycorrhizal seedlings is the number of the ectomycorrhizal seedlings.
Examples 1-6 the results of the pinus massoniana seed germination test are shown in fig. 1; the results of the pinus massoniana seed germination test of comparative examples 1-5 are shown in fig. 2.
FIG. 1 statistical chart of germination of pinus massoniana seeds coated with ectomycorrhizal fungi of examples 1-6 the germination percentage, average seedling height and mycorrhizal seedling ratio of the seeds of example 2 were the highest by comparing the results of the pinus massoniana seed germination test (as shown in FIG. 1) of the ectomycorrhizal fungi seed coating agent of examples 1-6; the germination condition of the masson pine seeds and the proportion of mycorrhizal seedlings after the coating of the ectomycorrhizal fungi are optimal under the formula ratio of 20L of ectomycorrhizal fungi liquid, 10Kg of water-retaining agent, 8Kg of adhesive, 40Kg of humus soil, 0.02Kg of gibberellin and 40Kg of pure water.
Fig. 2 comparative examples 1-5 germination conditions of ectomycorrhizal fungi coated masson pine seeds statistical figure 2 the results of comparative example 1 show that when the coating agent formula does not contain ectomycorrhizal fungi liquid, the seeds in the coating agent are not infected by ectomycorrhizal fungi, the germination parameters and lateral roots of the seeds are obviously reduced, and mycorrhizal seedlings cannot be formed; when the formula of the comparative example 2 does not contain the water-retaining agent, the seeds can not obtain sufficient water in the drought environment, and the germination rate is obviously reduced; comparative examples 3 to 5 in the absence of a tackifier, humus and gibberellin, respectively, the germination potential of the seeds is significantly reduced due to insufficient nutrient supply in the environment for seed germination such as insufficient organic matter and nutrient elements, or due to water hormone deficiency, thereby affecting the formation of ectomycorrhiza, resulting in a lower mycorrhizal shoot rate.
In addition, the invention can be used for direct seeding forestation on difficult lands. When the method is used, the masson pine seeds wrapped with the mycoderm agent are sown on forestation land, after the seeds germinate, the seeds are directly planted in the forestation land, and then mycorrhizal seedlings with good root system development and growth are formed, so that the survival of the seedlings can be effectively ensured. Meanwhile, the invention can avoid the problems of fine root damage caused by seedling transplantation during afforestation and long seedling recovery period after planting.
It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Further, it should also be understood that various alterations, modifications and/or variations can be made to the present invention by those skilled in the art after reading the technical content of the present invention, and all such equivalents fall within the protective scope defined by the claims of the present application.
It will be appreciated by those skilled in the art that the invention can be embodied in other specific forms without departing from the spirit or essential characteristics thereof. Thus, the embodiments disclosed above are illustrative and not exclusive in all respects. All changes which come within the scope of or equivalence to the invention are intended to be embraced therein.
Claims (10)
1. The masson pine ectomycorrhizal fungi seed coating agent is characterized by comprising:
2. the masson pine ectomycorrhizal fungi seed coating agent of claim 1, which is characterized in that: 20 parts of mycorrhizal fungi liquid, 10 parts of water-retaining agent, 8kg of adhesive, 40 parts of humus, 0.02 part of gibberellin and 40 parts of purified water.
3. The masson pine ectomycorrhizal fungi seed coating agent of claim 1, which is characterized in that: the ectobacteria are Boletus viscosus, Lasiosphaera leucocephala and Lactarius deliciosus.
4. The masson pine ectomycorrhizal fungi seed coating agent of claim 1 or 2, which is characterized in that: the preparation process of the boletus mucosae, the columba esculenta and the lactarius deliciosus comprises the following steps: respectively filling the three bacteria into a sterilized triangular flask containing 500ML culture solution, and then pricking the bottle mouth with 6 layers of sterile gauze; culturing at 25 deg.C with oscillation amplitude of 8cm and frequency of 120 times/min for 7 days to obtain 3 kinds of dense mixed fungi mycelia.
5. The masson pine ectomycorrhizal fungi seed coating agent of claim 1, which is characterized in that: the water-retaining agent is polyvinyl alcohol and water-absorbing resin SAP.
6. The masson pine ectomycorrhizal fungi seed coating agent of claim 1, which is characterized in that: the adhesive is water-soluble polyvinyl alcohol.
7. The masson pine ectomycorrhizal fungi seed coating agent of claim 1, which is characterized in that: the growth regulator is gibberellin.
8. A process for preparing a masson pine ectomycorrhizal fungi seed coating agent of any one of claims 1-5, comprising the steps of:
1) the preparation process of the boletus mucosae, the columba esculenta and the lactarius deliciosus comprises the following steps: respectively filling the three bacteria into a sterilized triangular flask containing 500ML culture solution, and then pricking the bottle mouth with 6 layers of sterile gauze; performing shaking culture at 25 deg.C with amplitude of 8cm and frequency of 120 times/min for 7 days to obtain 3 kinds of dense mixed fungi mycelia in liquid;
2) mixing mycelium liquid containing Boletus viscosus, Lasiosphaera leucocephala and Lactarius deliciosus, humus, gibberellin and viscosity agent, stirring, slowly adding purified water, stirring, stopping adding water, and standing to obtain a solidified state.
9. The method of using a masson pine ectotrophic seed coating formulation of any one of claims 1 to 7, characterized by comprising the steps of:
1) seed disinfection: preparing 2L of 600 times of liquid by using 75% chlorothalonil wettable powder with good adhesion, broad-spectrum sterilization and low toxicity, soaking seeds for 1 hour, and washing the seeds for later use by using distilled water for 3 times;
2) preparing a seed coating agent: mixing mycelium liquid containing Boletus viscosus, Lasiosphaera leucocephala and Lactarius deliciosus, humus, gibberellin and mucoadhesive, stirring, slowly adding purified water, stirring, stopping adding water, and standing to obtain a solidified mixture;
3) seed coating: the seeds for standby disinfection and the mixed seed coating agent are put into a seed coating machine for coating, and the mass ratio of the seed coating agent to the sterile seeds is (0.2-2): 1.
10. The masson pine ectomycorrhizal fungi seed coating agent of claim 1, which is characterized in that: the step 3) seed coating: and putting the disinfected seeds and the mixed seed coating agent into a seed coating machine for coating, wherein the mass ratio of the seed coating agent to the seeds is 2: 1.
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| CN108112298A (en) * | 2017-12-20 | 2018-06-05 | 西南大学 | A kind of breeding method of Mycorrhizal Fungi of Pinus massoniana nursery stock |
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| CN110754483A (en) * | 2019-10-29 | 2020-02-07 | 贵州大学 | Pinus massoniana seed coating agent and preparation method and application thereof |
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| CN108112298A (en) * | 2017-12-20 | 2018-06-05 | 西南大学 | A kind of breeding method of Mycorrhizal Fungi of Pinus massoniana nursery stock |
| CN110268917A (en) * | 2019-08-07 | 2019-09-24 | 西南大学 | Applying Ectomycorrhizal Fungi composite bacteria agent |
| CN110754483A (en) * | 2019-10-29 | 2020-02-07 | 贵州大学 | Pinus massoniana seed coating agent and preparation method and application thereof |
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