Mycorrhizal fungus organic carrier as well as preparation method and application thereof

Abstract

The invention provides a mycorrhizal fungus organic carrier as well as a preparation method and application thereof, and belongs to the technical field of biological agriculture. The mycorrhizal fungus organic carrier comprises calcium alginate hydrogel and an organic compound embedded in the calcium alginate hydrogel; the organic compound comprises a matrix and a carbon source; the concentration of the matrix in the organic compound is 8-45 g / L; the concentration of the carbon source is 23-72.5 g / L. The mycorrhizal fungi organic carrier can provide appropriate humidity, nutrition and attachment sites and a microenvironment suitable for growth for mycorrhizal fungi, and the mycorrhizal fungi with high activity and high rejuvenation speed can be obtained by inoculating the mycorrhizal fungi to the mycorrhizal fungi organic carrier. The mycorrhizal fungi resources can be effectively applied to production practice of the orchid plants, a stable symbiotic relationship with the orchid plants is established, growth of the orchid plants is promoted, and the mycorrhizal fungi have important significance on breeding and protection of the orchid plants and development of the orchid industry.

Description

Technical Field

[0001] The invention belongs to the technical field of biological agriculture, and in particular relates to a mycorrhizal fungus organic carrier and a preparation method and application thereof. Background Art

[0002] In the natural environment, the distribution and population dynamics of some plants are often closely related to the presence of suitable mycorrhizal fungi. Mycorrhizal fungi benefit plants mainly by enhancing their nutrient acquisition and stress tolerance. The seeds of orchids are as small as dust, contain almost no endosperm, lack sufficient nutrient reserves, and are highly dependent on the symbiotic relationship with orchid mycorrhizal fungi to obtain the carbon source and other nutrients required for germination and early growth. Orchid mycorrhizal fungi invade the cortical cells of seeds or roots through hyphae, forming a unique mycelium structure, and release carbon, nitrogen, phosphorus and other nutrients required for growth by decomposing organic matter in the matrix for orchids to absorb and utilize. So far, researchers have isolated and cultured a large number of orchid mycorrhizal fungi from the roots or protocorms of many orchids through various means. Through a large number of indoor symbiotic experiments, it has been shown that some specific mycorrhizal fungi can significantly promote the germination of orchid seeds and plant growth, and play a positive role in promoting the growth of orchids.

[0003] However, it is not easy to expand the culture of orchid mycorrhizal fungi in production because they cannot produce spores. The cultivation of mycorrhizal fungi in the laboratory is expanded by mycelium. After the mycelium is transferred to the cultivation medium, the activity is low and it cannot quickly recover. It cannot quickly and effectively form a symbiotic relationship with orchid plants, and thus cannot effectively play a role in promoting growth. Summary of the invention

[0004] In view of this, the present invention provides an organic carrier for mycorrhizal fungi, which can provide a suitable carrier environment for mycorrhizal fungi and promote their reproduction, expand culture, achieve rapid colonization, and thus achieve the purpose of promoting the growth of plants by mycorrhizal fungi.

[0005] In order to achieve the above object, the present invention provides the following technical solutions:

[0006] The present invention provides a mycorrhizal fungus organic carrier, comprising a calcium alginate hydrogel and an organic complex embedded in the calcium alginate hydrogel;

[0007] The organic complex includes a substrate and a carbon source;

[0008] The concentration of the matrix in the organic complex is 8 to 45 g / L;

[0009] The concentration of the carbon source is 23-72.5 g / L.

[0010] Preferably, the matrix comprises bentonite;

[0011] The carbon source includes starch and monosaccharides;

[0012] The concentration of the starch is 15-60 g / L;

[0013] The concentration of the monosaccharide is 8-12.5 g / L.

[0014] The present invention provides a method for preparing the mycorrhizal fungus organic carrier, comprising the following steps:

[0015] dissolving the substrate, the carbon source and sodium alginate in water to obtain a sodium alginate mixed solution;

[0016] The sodium alginate mixed solution is dropped into a calcium chloride aqueous solution for solidification reaction to obtain a mycorrhizal fungus organic carrier.

[0017] Preferably, the concentration of the matrix in the sodium alginate mixture is 8-45 g / L; the concentration of the sodium alginate is 5-10 g / L;

[0018] When the carbon source includes starch and monosaccharide, the concentration of the starch is 15-60 g / L, and the concentration of the monosaccharide is 8-12.5 g / L.

[0019] Preferably, the mass percentage of the calcium chloride aqueous solution is 1.8% to 2.2%.

[0020] The invention provides an organic carrier with mycorrhizal fungi attached thereto, which is obtained by inoculating mycorrhizal fungi on the mycorrhizal fungi organic carrier or the mycorrhizal fungi organic carrier prepared by the preparation method and culturing the same.

[0021] Preferably, the mycorrhizal fungi include at least one mycorrhizal fungi from the following families: Tulasnellaceae, Sebacinales and Ceratobasidiaceae;

[0022] The culture is dark culture, and the culture temperature is 23-27°C.

[0023] The invention provides the mycorrhizal fungus organic carrier, the mycorrhizal fungus organic carrier prepared by the preparation method, and the use of the organic carrier with mycorrhizal fungi attached thereto in the propagation of orchid plants.

[0024] Preferably, the organic carrier with mycorrhizal fungi attached thereto and the cultivation substrate are used together as a propagation substrate for sowing orchid seeds and / or cultivating orchids.

[0025] Preferably, the orchidaceae plant includes epiphytic orchids.

[0026] Compared with the prior art, the present invention has the following advantages:

[0027] The present invention provides an organic carrier for mycorrhizal fungi, comprising a calcium alginate hydrogel and an organic complex embedded in the calcium alginate hydrogel; the organic complex comprises a matrix and a carbon source; the concentration of the matrix in the organic complex is 8 to 45 g / L; the concentration of the carbon source is 23 to 72.5 g / L. The organic carrier for mycorrhizal fungi of the present invention rapidly absorbs moisture from the environment through the action of the components of the matrix, the carbon source and the calcium alginate hydrogel, providing suitable environmental humidity and nutrition for the growth of mycorrhizal fungi, and the special structure of the organic carrier for mycorrhizal fungi provides an attachment site and a suitable microenvironment for the growth of mycorrhizal fungi. Mycorrhizal fungi are inoculated onto the organic carrier for mycorrhizal fungi, and the organic carrier for mycorrhizal fungi creates moist living conditions for mycorrhizal fungi, maintains the activity of mycorrhizal fungi, enables them to rapidly rejuvenate and expand, and enhances the activity of mycorrhizal fungi in the cultivation matrix, which is conducive to effectively applying orchid mycorrhizal fungi resources to the production practice of orchid plants, establishing a stable symbiotic relationship with orchid plants, and promoting the growth of orchid plants.

[0028] Furthermore, the mycorrhizal fungi organic carrier of the present invention specifically defines that the matrix includes bentonite. The mycorrhizal fungi organic carrier produced using bentonite has a higher water absorption rate, and the volume can expand rapidly after absorbing water, providing a suitable carrier platform for fungal growth, which is more conducive to the rejuvenation and expansion of orchid mycorrhizal fungi. In a dry environment, bentonite can absorb moisture in the environment and continuously provide a moist growth environment for fungi.

[0029] The present invention provides an organic carrier with mycorrhizal fungi attached thereto, and based on the performance of the mycorrhizal fungi organic carrier in promoting the rejuvenation of mycorrhizal fungi, mycorrhizal fungi are inoculated on the mycorrhizal fungi organic carrier and cultured. After inoculation, the mycorrhizal fungi can grow rapidly on the mycorrhizal fungi organic carrier, completely covering the surface of the mycorrhizal fungi organic carrier to form an organic carrier with mycorrhizal fungi attached thereto. When the organic carrier with mycorrhizal fungi attached thereto is dried and then placed in a culture medium, the mycorrhizal fungi can be rapidly rejuvenated, thereby obtaining a large number of mycorrhizal fungi with high activity, which is beneficial for the mycorrhizal fungi to establish a symbiotic relationship with orchid plants, thereby playing a growth-promoting role.

[0030] The present invention provides an application of the mycorrhizal fungus organic carrier or the organic carrier with mycorrhizal fungi attached in the propagation of orchid plants. The organic carrier with mycorrhizal fungi attached can promote the growth and rejuvenation of dry mycorrhizal fungi, which is beneficial for mycorrhizal fungi to quickly establish a symbiotic relationship with orchid plants, thereby playing a growth-promoting role. In one embodiment of the present invention, the effects of the mycorrhizal fungus agar mixture and the organic carrier with mycorrhizal fungi attached to the present invention on the growth of orchid seedlings were compared. The results showed that the various growth indicators of the organic carrier treatment group with mycorrhizal fungi attached to the present invention were significantly higher than those of the mycorrhizal fungus agar mixture treatment group, especially in the number of roots, dry weight, plant height and leaf area of ​​orchid seedlings. It can be seen that the organic carrier with mycorrhizal fungi attached to the present invention can be used to promote the propagation of orchid plants, which is of great significance for the breeding and protection of orchid plants and the development of the orchid industry. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 Flow chart for the preparation of organic carriers for mycorrhizal fungi;

[0032] Figure 2 These are the moisture content, water absorption rate and fungal rejuvenation results of the organic carriers with mycorrhizal fungi attached to PF1~PF12, where a is the moisture content result, b is the water absorption rate result after drying, c is the rejuvenation growth rate of mycorrhizal fungi after drying and storage for 30 days, and d is the rejuvenation morphology of mycorrhizal fungi and the sample of the organic carrier with mycorrhizal fungi attached after drying. DETAILED DESCRIPTION

[0033] The invention provides a mycorrhizal fungus organic carrier, comprising a calcium alginate hydrogel and an organic complex embedded in the calcium alginate hydrogel; the organic complex comprises a matrix and a carbon source; the concentration of the matrix in the organic complex is 8-45 g / L; and the concentration of the carbon source is 23-72.5 g / L.

[0034] The mycorrhizal fungi organic carrier of the present invention rapidly absorbs environmental moisture through the action of the components of the matrix, the carbon source and the calcium alginate hydrogel, providing suitable environmental humidity and nutrition for the growth of mycorrhizal fungi, and the special structure of the mycorrhizal fungi organic carrier provides an attachment site and a suitable microenvironment for the growth of mycorrhizal fungi. Inoculating mycorrhizal fungi onto the mycorrhizal fungi organic carrier can obtain mycorrhizal fungi with high activity and fast rejuvenation speed, which is conducive to the establishment of a symbiotic relationship between mycorrhizal fungi and orchid plants, and promotes the growth of orchid plants.

[0035] In the present invention, the concentration of the matrix can be 10g / L, 15g / L, 20g / L, 25g / L, 30g / L, 35g / L or 40g / L. The matrix preferably includes bentonite. The bentonite is a non-metallic mineral with montmorillonite as the main mineral component. The bentonite swells when it meets water, and the volume expands several times to 20 to 30 times after absorbing water. The bentonite is preferably sodium bentonite, which has a high water absorption rate and a large expansion multiple. The organic carrier developed and produced using sodium bentonite has a higher water absorption rate, and the volume can expand rapidly after absorbing water, providing a suitable carrier platform for fungal growth, which is more conducive to the rejuvenation and expansion of orchid mycorrhizal fungi. In a dry environment, bentonite can absorb moisture in the environment and continuously provide a moist growth environment for fungi.

[0036] In the present invention, the concentration of the carbon source can be 27g / L, 35g / L, 38g / L, 41g / L, 55g / L, 65g / L or 70g / L. The carbon source is preferably a nutrient that provides energy and carbon source for the growth of orchid mycorrhizal fungi. The carbon source preferably includes starch and monosaccharides. The starch preferably includes corn starch and / or potato starch, and the potato starch is preferably extracted by boiling potato slices with water, and the boiling time is preferably 15min. The mass volume ratio of the potato to water is 100g:1L. The concentration of the starch is preferably 15-60g / L, and can be 20g / L, 25g / L, 30g / L, 35g / L or 40g / L, 45g / L or 55g / L. The monosaccharide preferably includes glucose. The concentration of the monosaccharide is preferably 8-12.5g / L, and can be 10g / L, 11g / L or 12g / L. The present invention does not specifically limit the purchase sources of corn starch, potato starch and glucose, and conventional commercial products in the art can be used. The calcium alginate hydrogel provides embedding materials for the organic complex and regulates the microenvironment of the mycorrhizal fungus organic carrier attachment site.

[0037] The present invention provides a method for preparing the mycorrhizal fungus organic carrier, comprising the following steps:

[0038] dissolving the substrate, the carbon source and sodium alginate in water to obtain a sodium alginate mixed solution;

[0039] The sodium alginate mixed solution is dropped into a calcium chloride aqueous solution for solidification reaction to obtain a mycorrhizal fungus organic carrier.

[0040] The invention dissolves the matrix, the carbon source and the sodium alginate in water to obtain the sodium alginate mixed solution.

[0041] The present invention does not specifically limit the dissolution method, and the conventional dissolution method in the art can be used. In an embodiment of the present invention, it is preferred that after adding a carbon source to water to dissolve, the matrix is ​​mixed for the first time, and sodium alginate is added after the first mixing to obtain a sodium alginate mixed solution. Mixing twice can fully mix the components, and the final addition of the carbon source will cause the carbon source to not be fully diffused and dissolved, resulting in an improper ratio of nutrients in the obtained mycorrhizal fungus organic carrier, which is not conducive to the growth of mycorrhizal fungi; and adding sodium alginate first and then bentonite will cause the mixed solution to solidify prematurely, the nutrients or matrix are not mixed evenly, and the organic carrier fails to be made. The mixing method preferably uses a magnetic stirrer to mix, and the magnetic stirrer can mix the components. The concentration of the matrix in the sodium alginate mixed solution is preferably 8-45g / L, and can be 10g / L, 15g / L, 20g / L, 25g / L, 30g / L, 35g / L or 40g / L. The concentration of sodium alginate is preferably 5-10g / L, and can be 6g / L, 7g / L, 8g / L or 9g / L; when the carbon source includes starch and monosaccharide, the concentration of starch is preferably 15-60g / L, and can be 20g / L, 25g / L, 30g / L, 35g / L or 40g / L, 45g / L or 55g / L; the concentration of monosaccharide is preferably 8-12.5g / L, and can be 10g / L, 11g / L or 12g / L. The pH of the sodium alginate mixture is preferably adjusted to 5.6-5.8, and can be 5.7. Adjusting the pH is conducive to the growth of mycorrhizal fungi.

[0042] After obtaining the sodium alginate mixed solution, the present invention drips the sodium alginate mixed solution into a calcium chloride aqueous solution for solidification reaction to obtain a mycorrhizal fungus organic carrier.

[0043] In the present invention, the method of dripping is preferably dripping through a 4-6 mm dripping bottle, more preferably a 5 mm dripping bottle. The mass percentage of the calcium chloride aqueous solution is preferably 1.8% to 2.2%, which can be 1.9%, 2.0% or 2.1%. The curing reaction is in the calcium chloride aqueous solution, the Na on the G unit (α-L-guluronic acid) + The ion exchange reaction occurs with divalent cations, and the G units are stacked to form a cross-linked network structure, thereby forming a hydrogel. The mycorrhizal fungus attachment carrier is spherical with a diameter of 0.5 to 0.8 mm.

[0044] The mycorrhizal fungi organic carrier of the present invention can provide a suitable microenvironment for the growth of mycorrhizal fungi, such as suitable humidity, nutrition and attachment sites. Mycorrhizal fungi can be inoculated into the mycorrhizal fungi organic carrier to obtain mycorrhizal fungi with high activity and fast rejuvenation speed, which is conducive to the establishment of a symbiotic relationship between mycorrhizal fungi and orchids and promotes the growth of orchids. In one embodiment of the present invention, the effect of a mixture formed by a combination of pine bark, water and carbon source on the growth of mycorrhizal fungi is compared. The results show that the mycorrhizal fungi can not grow when the mycorrhizal fungi are directly inoculated on the mixture, and the mycorrhizal fungi hyphae grow very slowly and weakly after being inoculated on the mixture. A small amount of hyphae can be seen after 20 days, and the mycorrhizal fungi are less active. The mycorrhizal fungi can grow rapidly on the mycorrhizal fungi organic carrier of the present invention, and a large amount of hyphae can be observed in 7 days. Continued cultivation can completely cover the mycorrhizal fungi organic carrier, and when placed in a culture medium after drying, the mycorrhizal fungi can be quickly rejuvenated. It can be seen that the mycorrhizal fungi organic carrier of the present invention can promote the growth of mycorrhizal fungi, improve the activity of mycorrhizal fungi, and is conducive to establishing a symbiotic relationship with orchids, thereby playing a growth-promoting role. In one embodiment of the present invention, the addition amount of starch (corn starch or potato starch), glucose, bentonite and sodium alginate is further used to prepare mycorrhizal fungi organic carriers PF1-PF12, and the moisture content, water absorption rate and fungal rejuvenation of different mycorrhizal fungi organic carriers after inoculation of mycorrhizal fungi are compared. The results show that PF2 (without bentonite) has lower moisture content and water absorption rate than the other groups, and the strain rejuvenation rate is slower. It can be seen that the moisture content, water absorption rate and strain rejuvenation rate of mycorrhizal fungi organic carriers after adding an appropriate amount of bentonite to docking bacteria play an important role. In addition, PF9 (including bentonite 10g, sodium alginate 8g) is compared with PF7 (including bentonite 8g, sodium alginate 8g) and PF11 (including bentonite 15g, sodium alginate 8g), PF9 moisture content and water absorption rate are significantly improved, and the fungal rejuvenation rate is accelerated, which shows the importance of the addition amount of bentonite and the ratio of bentonite and sodium alginate for the performance of mycorrhizal fungi organic carriers.

[0045] Based on the performance of the mycorrhizal fungi organic carrier in promoting the rejuvenation of mycorrhizal fungi, the present invention provides an organic carrier with mycorrhizal fungi attached thereto, which is obtained by inoculating mycorrhizal fungi on the mycorrhizal fungi organic carrier or the mycorrhizal fungi organic carrier prepared by the preparation method. The mycorrhizal fungi in the organic carrier with mycorrhizal fungi attached thereto can be rapidly rejuvenated, which is conducive to the establishment of a symbiotic relationship between mycorrhizal fungi and orchid plants, and promotes the growth of orchid plants.

[0046] The mycorrhizal fungi preferably include at least one mycorrhizal fungi from the following families: Tulasnellaceae, Sebacinales and Ceratobasidiaceae. The mycorrhizal fungi are preferably orchid mycorrhizal fungi that can form a symbiotic relationship with orchid plants and promote the germination and growth of orchid plants.

[0047] In the embodiment of the present invention, the mycorrhizal fungi used are TP-2 strain, TP-8 strain, TP-3 strain and TP-11 strain, which are strains disclosed in the prior art, and are disclosed in the patents of a kind of film fungus strain TP-2 and application for promoting the thick stem of Dendrobium (CN114395485B), a film fungus strain TP-8 and application thereof for improving the sprouting ability of Dendrobium seedlings (CN114381379B), a film fungus strain TP-3 and application thereof for promoting the high growth ability of Dendrobium plant (CN114395486B) and a tuber mycorrhizal fungus strain TP-11 and application thereof for promoting the growth of new leaves of Dendrobium (CN114507618B). The inoculation is preferably inoculated on the mycorrhizal fungus organic carrier in the form of a mycorrhizal fungus block or hyphae. The present invention does not specifically limit the preparation method of the fungus block or hyphae, and the fungus culture method commonly used in the art can be used. For example, a fungus block or hyphae is obtained by culturing in potato dextrose agar medium (PDA) or in potato dextrose medium. The mycorrhizal fungus organic carrier is preferably a sterilized mycorrhizal fungus organic carrier. The sterilization temperature is preferably 121°C, and the sterilization time is preferably 30 to 60 minutes, more preferably 30 minutes. During the inoculation, the mycorrhizal fungus organic carrier is inoculated into the moist mycorrhizal fungus organic carrier to improve the inoculation survival rate of the mycorrhizal fungus. The culture is preferably dark culture. The temperature of the dark culture is preferably 23 to 27°C, which can be 24°C, 25°C or 26°C. After 25 to 30 days of the culture, the mycorrhizal fungus hyphae cover the surface of the mycorrhizal fungus organic carrier, and an organic carrier with mycorrhizal fungi attached is obtained. In an embodiment of the present invention, a bottle of 300 mL of mycorrhizal fungus organic carrier is cultured for about 30 days to obtain an organic carrier with mycorrhizal fungi attached. The storage method of the organic carrier with mycorrhizal fungi attached is preferably to be dried by blowing on a clean bench and then sealed and stored at 4°C. The organic carrier with mycorrhizal fungi attached to it contains a large amount of mycorrhizal fungi with high activity and fast rejuvenation. When placed in an orchid cultivation environment, the mycorrhizal fungi can be rapidly rejuvenated, which is beneficial to the establishment of a symbiotic relationship between the mycorrhizal fungi and the orchid, and promotes the growth of the orchid.

[0048] The present invention provides application of the mycorrhizal fungi organic carrier, the mycorrhizal fungi organic carrier prepared by the preparation method or the organic carrier with mycorrhizal fungi attached thereto in orchid plant propagation.

[0049] In the present invention, it is preferred that the organic carrier with mycorrhizal fungi attached and the cultivation substrate are used together as a propagation substrate for sowing orchid seeds and / or cultivating orchids. The cultivation substrate preferably includes pine bark and / or sawdust. The length of the pine bark is preferably 6 to 9 mm. The volume ratio of the organic carrier with mycorrhizal fungi attached and the cultivation substrate is preferably 1:20. The method of using them together as a propagation substrate preferably comprises mixing the organic carrier with mycorrhizal fungi attached and the cultivation substrate. The present invention does not particularly limit the mixing method, and a conventional mixing method in the art can be used. In an embodiment of the present invention, the organic carrier with mycorrhizal fungi attached is evenly sprinkled on the surface of the cultivation substrate. The orchid preferably includes an epiphytic orchid, specifically Dendrobium officinale.

[0050] In one embodiment of the present invention, the effects of the mycorrhizal fungi agar mixture and the organic carrier with mycorrhizal fungi attached to the present invention on the growth of orchid seedlings were compared, and the results showed that the growth indexes of the organic carrier with mycorrhizal fungi attached to the present invention were significantly higher than those of the mycorrhizal fungi agar mixture treatment group, especially in the number of roots, dry weight, plant height and leaf area of ​​orchid seedlings. It can be seen that the organic carrier with mycorrhizal fungi attached to the present invention can be used to promote the reproduction of orchids.

[0051] In order to further illustrate the present invention, a mycorrhizal fungus organic carrier provided by the present invention and its preparation method and application are described in detail below in conjunction with the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

[0052] Example 1

[0053] Preparation method of mycorrhizal fungi organic carrier (potato starch)

[0054] Weigh 100 g of fresh potatoes, heat and cook for 15 minutes, filter out the potatoes, retain the filtrate, and obtain 1 L of solution A; add 8 g of glucose to solution A, stir and dissolve evenly to obtain a mixed solution B; slowly add 25 g of bentonite to the mixed solution B, use a magnetic stirrer to stir continuously and fully to accelerate the dissolution, and obtain a mixed solution C; slowly add 10 g of sodium alginate to the mixed solution C, use a magnetic stirrer to stir continuously to make all the raw materials fully dissolved and mixed evenly, and adjust the pH to 5.6-5.8 to obtain a mixed solution D;

[0055] Prepare 2%w CaCl2 solution to obtain solution F; fill the mixed solution D into a dropper bottle with a mouth diameter of 5mm, and then drop the mixed solution D into solution F. The mixed solution D quickly gels into balls, and the mycorrhizal fungal organic carrier can be obtained, which is recorded as PF1.

[0056] Comparative Example 1

[0057] Preparation method of mycorrhizal fungi organic carrier (without bentonite)

[0058] Weigh 30g corn starch and dissolve it in 1L water to obtain solution A; add 8g glucose to solution A, stir and dissolve to obtain mixed solution B; stir and dissolve to obtain mixed solution B; slowly add 8g sodium alginate to mixed solution B, stir continuously with a magnetic stirrer to make all raw materials fully dissolved and mixed, and adjust the pH to 5.6-5.8 to obtain mixed solution D;

[0059] Prepare 2%w CaCl2 solution to obtain solution F; fill the mixed solution D into a dropper bottle with a mouth diameter of 5mm, and then drop the mixed solution D into solution F. The mixed solution D quickly gels into balls, and the mycorrhizal fungal organic carrier can be obtained, which is recorded as PF2.

[0060] Example 2

[0061] Preparation method of mycorrhizal fungus organic carrier

[0062] Weigh 15g corn starch and dissolve it in 1L water to obtain solution A; add 8g glucose to solution A, stir and dissolve to obtain mixed solution B; stir and dissolve to obtain mixed solution B; slowly add 12.5g bentonite to mixed solution B, stir and dissolve with a magnetic stirrer to accelerate dissolution, and obtain mixed solution C; slowly add 10g sodium alginate to mixed solution C, stir and dissolve with a magnetic stirrer to make all raw materials fully dissolved and mixed, and adjust the pH to 5.6-5.8 to obtain mixed solution D;

[0063] Prepare 2%w CaCl2 solution to obtain solution F; fill the mixed solution D into a dropper bottle with a mouth diameter of 5mm, and then drop the mixed solution D into solution F. The mixed solution D quickly gels into balls, and the mycorrhizal fungal organic carrier can be obtained, which is recorded as PF3.

[0064] Comparative Example 2

[0065] Preparation method of mycorrhizal fungi organic carrier (added MMN culture medium)

[0066] Weigh 10g of MMN culture medium, add it to 1L of water, use a magnetic stirrer to stir and dissolve it, then weigh 10g of bentonite, slowly add it to the MMN solution, use a magnetic stirrer to stir and mix it evenly, and finally weigh 5g of sodium alginate, slowly add it to the mixed solution. As a result, the mixed solution gradually forms a gel from top to bottom, and it is impossible to make an organic carrier with distinct particles.

[0067] Example 3

[0068] Based on the preparation method of the mycorrhizal fungal organic carrier in Example 2, the addition amounts of corn starch, glucose, bentonite and sodium alginate were adjusted to prepare PF4-PF12 mycorrhizal fungal organic carriers. The specific addition amounts are shown in Table 1.

[0069] Table 1 Content of each component of PF4~PF12 mycorrhizal fungal organic carrier

[0070] Group Corn starch glucose Bentonite Sodium alginate PF4 15 12 30 5 PF5 30 5 20 6 PF6 30 11 10 10 PF7 30 8 8 8 PF8 30 8 15 6 PF9 30 8 10 8 PF10 30 8 10 5 PF11 30 8 15 8 PF12 60 10 45 10

[0071] Example 5

[0072] Moisture content, water absorption rate and fungal rejuvenation of organic carriers with mycorrhizal fungi attached to PF1-PF12 Preparation of organic carriers with mycorrhizal fungi attached to PF1-PF12: Put the organic carriers with mycorrhizal fungi attached to PF1-PF12 into tissue culture bottles, sterilize them at 121°C for 30 minutes, move them to the clean bench, and cool them to obtain moist organic carriers with mycorrhizal fungi; use a sterile inoculation loop to pick the bacterial blocks of TP-8 strains, inoculate them into the tissue culture bottles with mycorrhizal fungi organic carriers, and cover them with bottle caps to prevent contamination; after the inoculation, place the tissue culture bottles in the dark at 25±2°C. When the fungi grow and attach to the organic carriers, PF1-PF12 organic carriers with mycorrhizal fungi attached to PF1-PF12 can be obtained. Each group will blow dry the organic carriers with mycorrhizal fungi attached to the clean bench. Put them into self-sealing bags and seal them, and store them at 4°C.

[0073] 50 organic carriers with mycorrhizal fungi attached were randomly selected from each group, and their weights were weighed using a balance, W0, and then placed on a clean bench to be blown dry until the weight was constant. Then their weights were weighed to obtain W0. t , calculate the moisture content (W) according to the following formula I.

[0074] W(%)=[(W0-W t ) / W0]×100%Formula I.

[0075] After drying and storing for 30 days, 50 dry organic carriers with mycorrhizal fungi attached were randomly taken out from each group, and the weight of each organic carrier was weighed S0. Then, the organic carriers were placed in water for 1 hour to fully absorb water, and the weight of each organic carrier was weighed S0. t , the water absorption (S) is calculated according to the following formula II.

[0076] S(%)=[(S t -S0) / S0]×100% formula II.

[0077] After 30 days of drying and storage, 5 dried organic carriers with mycorrhizal fungi attached were randomly taken out from PF2, PF9 and PF11 to detect the rejuvenation of the fungi. Specifically, the dried organic carriers with mycorrhizal fungi attached were placed in PDA culture medium for culturing to rejuvenate the attached fungal hyphae. The radius of the fungal colony was measured and recorded every day to indicate the rejuvenation rate of the fungi.

[0078] The organic carrier with mycorrhizal fungi attached to PF5 did not successfully form a granular carrier, and its water content and water absorption rate could not be accurately tested. The water content, water absorption rate and fungal rejuvenation of the organic carrier with mycorrhizal fungi attached to PF1~PF4 and PF6~PF12 are shown in Figure 2. Figure 2 As shown. Figure 2 It can be seen that the moisture content, water absorption rate and fungal rejuvenation performance of organic carriers attached with mycorrhizal fungi in PF1~PF4 and PF6~PF12 are good. Specifically, the PF1 group has the highest moisture content, up to 85.4%, but the water absorption rate is less than 50%. The moisture content, water absorption rate and fungal rejuvenation rate of PF2 are significantly lower than those of PF3~PF4 and PF6~PF12. PF9 has a higher moisture content and the highest water absorption rate, which can reach 173%, and the fungal rejuvenation rate is the fastest.

[0079] The formula of PF9 was finally determined after many improvements in the proportion of raw materials. Compared with PF7 and PF11, the amount of bentonite in PF9 is different. The amount of bentonite in PF7 and PF11 is 8g and 15g respectively. Although the difference in moisture content is not significant, after the bentonite content is changed to 10g of PF9, the water absorption rate and fungal rejuvenation rate are significantly higher than those of PF7 and PF11. Compared with PF2, PF9 adds 15g of bentonite, which significantly improves the moisture content, water absorption rate and speeds up the fungal rejuvenation rate of PF9, and also further confirms the importance of appropriate amount of bentonite to the performance of mycorrhizal fungal organic carriers. Therefore, the method of PF9 will be used to mass-produce a large number of mycorrhizal fungal organic carriers and use them in the cultivation application of orchid plants.

[0080] Comparative Example 3

[0081] In order to screen suitable culture carriers for orchid mycorrhizal fungi, the following three mixtures were prepared with pine bark as material: mixture 1 was 30 m of pine bark with a length of 9 mm and 5 ml of water; mixture 2 was 30 ml of pine bark with a length of 9 mm and 10 ml of water; mixture 3 was 30 ml of pine bark with a length of 9 mm, 10 ml of water and 2 g of glucose.

[0082] Mixtures 1 to 3 were placed in tissue culture bottles, sterilized by high pressure for 60 min, cooled, and inoculated with 5 1 cm 3 Orchidaceous mycorrhizal fungus clumps (TP-8), cultured in the dark.

[0083] After 10 days of cultivation, the fungus blocks in mixture 1 dried up and turned brown-yellow, and no new mycelium grew. The fungus blocks in mixtures 2 and 3 did not dry up, but also failed to grow new mycelium. After 15 days of cultivation, the fungus blocks in mixture 3 swelled, the substrate fermented, and also failed to grow new mycelium.

[0084] Another inoculation method was used for cultivation: the mixture 3 was placed in a tissue culture bottle, sterilized by high pressure for 60 minutes, cooled, and inoculated with 1g of dried pure mycelium of orchid mycorrhizal fungi (TP-8). After culturing in the dark for 10 days, the mycelium was found to be swollen, but no new mycelium was found. After 20 days, only a few new mycelium were observed. Although this method can grow mycelium, it is very time-consuming and requires a lot of time and consumables. It is not convenient to use a shaker to cultivate mycelium, and the fungal activity is low, which is not suitable for widespread application in production practice.

[0085] Example 6

[0086] Application of organic carriers with mycorrhizal fungi in the cultivation of Dendrobium officinale

[0087] 1. Preparation of mycorrhizal fungi agar powder: Inoculate the bacterial mass of TP-8 strain into potato dextrose liquid medium (PDB) and culture by shaking. When the mycelium fills the container, filter and dry to obtain dry pure mycelium. Mix the dry pure mycelium with agar powder in a mass ratio of 1:5 and crush into powder.

[0088] Preparation of organic carrier with mycorrhizal fungi attached: The method for preparing organic carrier of mycorrhizal fungi of PF9 in Example 5 is the same, except that the orchid mycorrhizal fungi blocks selected during inoculation are blocks of TP-8 strain.

[0089] Preparation of mixed mycorrhizal fungal agar powder: Inoculate the bacterial blocks of TP-2 strain, TP-3 strain and TP-11 strain into PDA liquid culture medium respectively and culture with shaking. When the mycelium fills the container, filter to obtain pure mycelium, dry, mix with agar powder and crush into powder to obtain powder of the three fungi, and then mix the three powders of equal mass evenly.

[0090] Preparation of mixed fungal mycorrhizal fungal organic carrier: The preparation method of the mycorrhizal fungal organic carrier of PF9 in Example 5 is the same, the only difference is that the fungal blocks of three fungi, namely TP-2 strain, TP-3 strain and TP-11 strain, are selected during inoculation.

[0091] 2. Cultivation of Dendrobium officinale seedlings

[0092] Treatment groups 1 to 5 were set up, each with 30 pots, and the specifications of the pots were 25×18×6.5 cm; 12 clumps were cultivated in each pot, and 4 Dendrobium officinale seedlings were cultivated in each clump. The number of leaves of each Dendrobium officinale seedling was 4 to 5, the number of roots was 2 to 3, the stem diameter was 1.5 to 1.8 mm, the longest root length was 2 cm, and the dry weight was 0.013 g. The cultivation medium was pine bark with a length of 6-9 mm.

[0093] Blank group: no fungus was applied;

[0094] Control group 1: 15 g of mycorrhizal fungus agar powder was evenly sprinkled on the surface of pine bark;

[0095] Treatment group 1: 15 g of mycorrhizal fungi organic carrier was evenly spread on the surface of pine bark;

[0096] Control group 2: 15 g of mixed mycorrhizal fungi agar powder was evenly sprinkled on the surface of pine bark;

[0097] Treatment group 2: 15 g of mixed fungal mycorrhizal fungal organic carrier was evenly spread on the surface of pine bark.

[0098] Each treatment group was cultured in a culture room and watered every 10 days. The samples were harvested after 6 months. Thirty-one samples of Dendrobium officinale were randomly selected from each treatment group. The number of leaves, the number of roots, the longest root length, the plant height (stem height), the leaf area and the dry weight of Dendrobium officinale were counted. The effects of different treatments on the growth of Dendrobium officinale were compared by one-way analysis of variance, and the mean values ​​of each treatment were compared by least significant difference (LSD), P<0.05.

[0099] The results of the analysis of the effects of different treatment groups on the growth of Dendrobium officinale are shown in Table 2.

[0100] Table 2 Comparison of growth indexes of Dendrobium officinale in different treatment groups

[0101] Treatment Group Blank Group Control group 1 Treatment Group 1 Control group 2 Treatment Group 2 Number of blades 5.45±0.29bB 7.42±0.25A 8.07±0.3A 7.37±0.23a 8.61±0.24a Number of roots 5.16±0.26cC 7.84±0.37B 9.13±0.45A 7.52±0.26b 9.45±0.29a Longest root length / cm 5.56±0.49bB 9.74±0.51A 9.99±0.39A 8.51±0.34a 10.57±0.49a Dry weight / g 0.06±0cC 0.1±0.01B 0.14±0.01A 0.11±0.01b 0.16±0.01a Plant height / cm 2.75±0.09cC 3.27±0.18B 3.75±0.19A 3.43±0.1b 4.01±0.2a <![CDATA[Leaf area / cm 2 > 1.46±0.07cC 2.06±0.09B 3.07±0.22A 2.08±0.1b 2.98±0.16a

[0102] Note: Different capital letters or lowercase letters indicate significant differences among different treatment groups.

[0103] The results in Table 2 show that compared with the blank group, the control groups 1-2 and the treatment groups 1-2 have a significant promoting effect on the growth of Dendrobium officinale, which is specifically manifested in that the number of leaves, number of roots, longest root length, plant height, leaf area and dry weight of Dendrobium officinale are significantly higher than those of the blank group. In addition, both the control group 1 and the treatment group 1 used the orchid mycorrhizal fungus TP-8, and the growth indicators of the treatment group 1 were higher than those of the control group 1, especially the number of roots, dry weight, plant height and leaf area of ​​Dendrobium officinale, which were significantly higher than those of the control group 1, which were 9.13±0.45, 0.14±0.01g, 3.75±0.19cm and 3.07±0.22cm respectively. 2Similarly, in both control group 2 and treatment group 2, three fungi, TP-2, TP-3, and TP-11, were used. However, the root number, dry weight, plant height, and leaf area of ​​Dendrobium officinale in treatment group 2 were significantly higher than those in control group 2 without the use of carrier, which were 9.45±0.29, 0.16±0.01g, 4.01±0.2cm, and 2.98±0.16cm, respectively. 2 .

[0104] The above results indicate that the organic carrier attached with mycorrhizal fungi of the present invention can effectively cultivate and propagate orchid mycorrhizal fungi, and further enhance the effect of fungi on promoting the growth of orchid plants.

[0105] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. Other embodiments can be obtained based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A mycorrhizal fungus organic carrier, characterized in that: It comprises a calcium alginate hydrogel and an organic complex embedded in the calcium alginate hydrogel; The organic complex includes a substrate and a carbon source; The concentration of the matrix in the organic complex is 8 to 45 g / L; The concentration of the carbon source is 23-72.5 g / L.

2. The mycorrhizal fungal organic carrier according to claim 1, characterized in that: The matrix includes bentonite; The carbon source includes starch and monosaccharides; The concentration of the starch is 15-60 g / L; The concentration of the monosaccharide is 8-12.5 g / L.

3. A method for preparing the mycorrhizal fungal organic carrier according to claim 1 or 2, characterized in that: The following steps are involved: dissolving the substrate, the carbon source and sodium alginate in water to obtain a sodium alginate mixed solution; The sodium alginate mixed solution is dropped into a calcium chloride aqueous solution for solidification reaction to obtain a mycorrhizal fungus organic carrier.

4. The preparation method according to claim 3, characterized in that: The concentration of the matrix in the sodium alginate mixed solution is 8-45 g / L; the concentration of the sodium alginate is 5-10 g / L; When the carbon source includes starch and monosaccharide, the concentration of the starch is 15-60 g / L, and the concentration of the monosaccharide is 8-12.5 g / L.

5. The preparation method according to claim 3 or 4, characterized in that: The mass percentage of the calcium chloride aqueous solution is 1.8% to 2.2%.

6. An organic carrier with mycorrhizal fungi attached, characterized in that: The fungus is inoculated and cultured on the fungus organic carrier according to claim 1 or 2 or the fungus organic carrier prepared by the preparation method according to any one of claims 3 to 5.

7. The organic carrier with mycorrhizal fungi attached thereto according to claim 6, characterized in that: The mycorrhizal fungi include at least one mycorrhizal fungi from the following families: Tulasnellaceae, Sebacinales, and Ceratobasidiaceae; The culture is dark culture, and the culture temperature is 23-27°C.

8. Use of the mycorrhizal fungal organic carrier according to claim 1 or 2, the mycorrhizal fungal organic carrier prepared by the preparation method according to any one of claims 3 to 5, or the organic carrier with mycorrhizal fungi attached thereto according to claim 6 or 7 in the propagation of orchid plants.

9. The use according to claim 8, characterized in that: The organic carrier with mycorrhizal fungi attached thereto and the cultivation substrate are used together as a propagation substrate for sowing orchid seeds and / or cultivating orchids.

10. The use according to claim 8 or 9, characterized in that: The orchidaceae plants include epiphytic orchids.

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