KR20140073129A - Method for manufacturing media of am fungi and am fungi media manufactured thereby - Google Patents

Abstract

The present invention relates to a method for preparing a carrier of vesicular arbuscular mycorrhizae, whereby biomass is used and waste resources are reutilized to achieve the optimization to the growing environment of vesicular arbuscular mycorrhizae, thereby easily mass-producing vesicular arbuscular mycorrhizae. The method comprises: a biomass preparation step of disrupting and screening biomass to remove impurities, followed by drying, thereby preparing biomass; a bio-charcoal preparation step of putting the prepared biomass in a sealed container, followed by indirect heating in the oxygen-free atmosphere to thermally decompose the biomass, thereby preparing bio-charcoal; a particle fractionation step of filtering the prepared bio-charcoal through a sieve while crushing the bio-charcoal, thereby fractionating into particles having four kinds of sizes, a first particle having a particle size of 2.0 mm to 3.0 mm, a second particle having a particle size of larger than 0.5 mm but smaller than 2.0 mm, a third particle having a particle size of 0.05 mm but smaller than 0.5 mm, and a fourth particle having a particle size of smaller than 0.05 mm; and a particle mix step of mixing the fractionated four kinds of particles at a ratio of 50-60 vol% of the first particle, 20-30 vol% of the second particle, 10-20 vol% of the third particle, and 10 vol% of the fourth particle.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a method for producing an endogenous mycaminocyte carrier and an endogenous mycorrhizal carrier prepared thereby,

The present invention relates to a method for producing an endogenous mycorrhizal microorganism used for culturing an endogenous mycorrhizae. More particularly, the present invention relates to a method for producing an endogenous mycorrhizal microorganism capable of culturing endemic mycorrhizae in a large amount And to an endogenous mycorrhizal carrier prepared by the method.

In general, mycorrhiza refers to roots in which the roots and fungi of higher plants are intimately joined together and have a symbiotic relationship. It is known that when plants lack moisture and nutrients in the soil, symbiosis with mycorrhiza is good for plant growth.

Mycorrhizae grows by using the photosynthetic products of the host plants after invading the roots of the host plants. The mycorrhizae grow in the host plants through the mycelium and the water parasites Specificity. As such, mycorrhiza is infected by mycorrhizae and maintains a symbiotic relationship with the host plant, and the nutrient and water absorption efficiency of the mycorrhiza plant is more than ten times higher than that of the uninfected root.

These mycorrhizas can be divided into endogenous mycorrhizal fungi that mycelium penetrates the cell wall of roots and forms branching hyphae in plant cells, and exogenous mycorrhiza that do not invade but proliferate in the cell wall of the cell layer. Endogenous mycorrhizas are distributed in nearly 100 species ranging from tropical tropics to polar regions around the world and can be found in all crops such as corn, cotton, wheat, potatoes, beans, alfalfa and sugarcane, . Arbuscular Mycorrhizal Fungi (AM Fungi) among these endogenous mycorrhizas is a typical endogenous mycorrhizae, and is a kind of mold which forms mycorrhiza together with host plants.

AM fungi inoculated to plants facilitates the absorption of water and nutrients in host plants by widening the contact area with soil through mycorrhiza and mycelium, especially in soybean plants. It also absorbs phosphorus, copper, and zinc, which have low fluidity in the soil, and supplies them to host plants. In particular, mycorrhiza is insoluble in the soil, so that it can absorb chemical fertilizer efficiently by absorbing and transferring the phosphoric acid which required artificially fertilization by mycelium.

In addition, AM fungi produce plant growth-promoting hormones such as Auxin, Cytokinin, and Gibberellin, promoting host plant growth, increasing crop yields, and at the same time improving soil quality , That is, the role of promoting the incorporation of soil.

Mycorrhiza, which is a soil microorganism, lives in various kinds of plant roots and absorbs nutrients and moisture in areas where roots are not accessible through hyphae occurring in infected plant roots and delivers them to plants. From the host plants, the carbon source needed for living places and metabolism is supplied to the soil, thereby improving the natural carbon cycle and soil physical, chemical and biological environment.

However, the conventional artificial cultivation of mycorrhizae as described above has the following problems.

Although mycorrhizae have many advantages as described above, they have not been put to practical use because the mycorrhizae bacteria themselves can live only by co-culturing with the roots of the host plant, Independent mass culture such as general microorganisms is impossible to date.

Therefore, there is an urgent need in the art for the development of a carrier capable of culturing mycorrhiza in a large amount without changing the growth characteristics of mycorrhizae that coexist with the host plant, and in particular, a carrier capable of culturing a large amount of endogenous mycorrhizae.

Published Patent No. 1998-0085224 "Vegetation composition for greening containing symbiotic bacteria" (Dec. 05, 1998)

SUMMARY OF THE INVENTION Accordingly, the present invention has been made keeping in mind the above problems occurring in the prior art,

An object of the present invention is to provide a method for manufacturing an endogenous mycorrhizal microorganism carrier capable of easily and massively cultivating endogenous mycorrhizal microorganisms by optimizing the growth environment of endemic mycorrhizae while recycling waste resources using biomass, and to provide an endogenous mycorrhizal carrier prepared thereby .

Another object of the present invention is to provide a method for manufacturing an endogenous mycorrhizal microorganism carrier, in which the biomass is dried at a moisture content of 20% or less to make the biomass pyrolyse smoothly and stably.

It is another object of the present invention to provide a method for manufacturing an endogenous mycorrhizal microorganism carrier in which the heating temperature of the biomass is limited to a certain extent so that the generation of biochar by thermal decomposition is more smoothly achieved.

It is another object of the present invention to provide a method for manufacturing an endogenous mycorrhizal microorganism carrier, which can optimize the growth environment of the endogenous mycorrhizal bacteria by limiting the pore size of the particles separated from the biochar.

In order to achieve the above object, a "method for producing an endogenous mycorrhizal microorganism carrier" according to the present invention comprises: a biomass preparation step of preparing biomass by removing impurities after crushing and screening biomass; Preparing biochar in which the biomass is contained in a closed container and indirectly heated in an oxygen-free atmosphere to pyrolyze the biomass to produce biochar; The produced biochars are sieved while being sieved, the first particles having a particle size of 2.0 mm to 3.0 mm, the second particles having a particle size larger than 0.5 mm and smaller than 2.0 mm, and a second particle having a particle size larger than 0.05 mm and 0.5 mm And a fourth particle having a particle size of less than 0.05 mm; By volume of the first particles, 50 to 60 volume% of the first particles, 20 to 30 volume% of the second particles, 10 to 20 volume% of the third particles, and 10 volume% A particle mixing step of mixing four particles; .

In the biomass preparation step of the "method for producing an endogenous mycorrhizal microorganism carrier" according to the present invention, the biomass is dried to a moisture content of 20% or less.

In the "method for producing an endogenous mycorrhizal microorganism carrier" according to the present invention, the heating temperature of the biomass is 350 ° C. to 450 ° C.

Further, in the above-described particle sorting step of " the method for producing endogenous mycorrhizal carrier according to the present invention, " the size of the pores of the first particles is larger than 50 m, the size of pores of the second particles is 10 m to 50 m, The size of the pores of the third particles is 5 占 퐉 to 10 占 퐉, and the size of the pores of the fourth particles is smaller than 5 占 퐉.

Further, in the biomass preparation step of the "method for producing an endogenous mycorrhizal microorganism carrier" according to the present invention, the biomass is a woody biomass.

INDUSTRIAL APPLICABILITY As described above, the present invention has an effect of easily cultivating endemic mycorrhizae in large quantities by optimizing the growth environment of endemic mycorrhizae while recycling waste resources using biomass.

Further, the present invention has the effect that the biomass is dried at a moisture content of 20% or less, and the biodegradable material is smoothly and thermally decomposed.

In addition, the present invention has the effect that the heating temperature of the biomass is limited to be constant, and the production of biochar by the thermal decomposition is more smoothly performed.

In addition, the present invention has the effect of optimizing the growth environment of the endogenous mycorrhizal bacteria by limiting the pore size of the particles separated from the biochar.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a diagram showing a method for producing an endogenous mycorrhizal carrier according to the present invention; FIG.

Hereinafter, preferred embodiments of the present invention will be described in detail. It should be understood, however, that the invention can be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a view showing a method for producing an endogenous mycorrhizal carrier according to the present invention; FIG.

As shown in the figure, the method for producing an endogenous mycorrhizal microorganism carrier according to the present invention comprises a biomass preparing step (S10) for preparing biomass, a biochar producing step (S20) for producing bio-char in the biomass, A particle sorting step (S30) for sorting particles by char size, and a particle mixing step (S40) for preparing an endogenous mycelial carrier by mixing the selected particles at a certain ratio.

In the biomass preparation step (S10), the biomass is crushed and screened to remove impurities, followed by drying to prepare biomass. This step is a process for preparing a material for producing bio-charcoal by crushing the lumpy biomass to a certain size or less, preferably 2 mm or less, screening the impurities with a sieve, and drying the biomass from which the impurities have been removed.

Here, the biomass is a renewable organic material extracted from energy-dedicated crops and trees, agricultural and feed crops, agricultural wastes and debris, forest wastes and debris, aquatic plants, animal waste, municipal wastes, and other wastes It refers to wood, plants, agricultural and forestry by-products currently used as energy sources, and organic components in municipal waste and industrial waste.

Among such biomass, the biomass suitable for the present invention is preferably a woody biomass suitable for the growth and cultivation of plants and hyphae. Examples of such woody biomass include bark, wood by-product, and sawdust.

In the biomass preparation step (S10), it is preferable that the biomass is dried at a moisture content of 20% or less. If moisture contained in the biomass is less than 20%, the bio- This is because pyrolysis of biomass can be promoted more smoothly. That is, if the moisture contained in the biomass is higher than 20%, the pyrolysis of the biomass due to the indirect heating is not performed smoothly and quickly.

In the biochar forming step (S20), the prepared biomass is housed in a closed container and indirectly heated in an oxygen-free atmosphere to pyrolyze the biomass to produce bio-char. In this step, the biomass is pyrolyzed in an anaerobic atmosphere to convert the biomass into the bio-char. The reason why the biomass is converted into the bio-char is that since the carrier base material itself according to the present invention is sterilized, To have a pore size of about 10 mu m on the surface of the substrate.

The indirect heating in the anoxic atmosphere is performed by discharging air from the inside of the closed container containing the biomass to form an oxygen free atmosphere and then heating the vessel itself or indirect heating such as high frequency heating To heat the biomass contained in the vessel.

The reason for heating in the anaerobic atmosphere is to prevent the biomass from being ignited by oxygen in the air while being heated. Here, the oxygen-free atmosphere means that oxygen is absent, but when such conditions can not be completely formed, heating can be performed even in a low-oxygen atmosphere, provided that ignition is minimized or hardly caused by oxygen. Therefore, it is preferable that the anaerobic atmosphere is strictly interpreted to include a low-oxygen atmosphere in addition to a completely anoxic atmosphere.

It is preferable that the heating temperature of the biomass is 350 ° C to 450 ° C in the biochar production step (S20). If the biomass is heated to a temperature lower than 350 ° C, the oil contained in the biomass is discharged to the outside, And if heated at a temperature higher than 450 캜, the biomass itself is heated to an excessively high temperature and carbonized before pyrolysis.

In the particle sorting step S30, the produced bio-char is sieved while being pulverized, so that the first particles having a particle size of 2.0 mm to 3.0 mm, the second particles having a particle size larger than 0.5 mm and smaller than 2.0 mm, A third particle having a size larger than 0.05 mm and smaller than 0.5 mm and a fourth particle having a particle size smaller than 0.05 mm. In this step, the produced biochar is pulverized and separated into a specific particle size and then sorted.

In the particle sorting step (S30), the size of the pores of the first particles is larger than 50 mu m, the size of pores of the second particles is 10 mu m to 50 mu m, And the size of the voids of the fourth particles is preferably smaller than 5 占 퐉.

This means, for example, that the pore size of the first particles is preferably greater than 50 占 퐉 in consideration of the size of the first particles and the shape of the particles, that is, as a result of measurement on a conventional particle size, When the size is 2.0 mm to 3.0 mm, it means that pores (voids formed between the particles and the particles) larger than 50 μm are usually formed. Thus, the second to fourth particles, as well as the first particles described for example, have a pore size according to the results obtained by conventional measurements, and the present invention limits the size of such pores, Clearly describe.

Generally, the size of mycelium of myogenetic mycorrhizae is 2 to 3 mu m (= 0.003 mm), the average length of mycelia is 2 to 15 mu m, and the size of spores of mycorrhizal mycorrhizae is about 50 mu m (= 0.05 mm) To produce pores capable of smoothly growing mycelium and spores of the first to fourth particles, so that the specific pores can be smoothly formed by mixing the particles.

The particle mixing step S40 may include mixing 50 to 60 volume% of the first particles, 20 to 30 volume% of the second particles, 10 to 20 volume% of the third particles, 10 volume% 4 < / RTI > particles. This step is a process for completing the carrier for endogenous mycorrhagic fungi according to the present invention by mixing the separated biochar in a specific ratio by particle. By mixing these particles at specific ratios, the size and ratio of the pores formed by the particles inside the carrier are adjusted to suit the hyphae of the endogenous mycorrhizae and the growth of the spores.

The specific mixing ratios of the particles forming the carrier according to the present invention as described above were obtained by experiments, and the process and results of such experiments are described below.

Experimental course

The process of comparing the infection rate of endogenous mycorrhizal bacteria to carriers for endogenous mycorrhizae prepared by mixing at a specific ratio will be described as follows.

In this experiment, two crops such as sorghum and Sudan grass and the following carrier components were tested and repeated five times for each example. The components of the carrier are shown in Table 1.

carrier  ingredient Example 1 Porosity-controlled biochar (invention) Comparative Example 1 Soils: Soils = 1: 1 (mixed soil and soil 1: 1) Comparative Example 2 Bio-charcoal Comparative Example 3 Activated carbon Comparative Example 4 Zeolite Comparative Example 5 Charcoal

Example 1

In Example 1, the infection rate of endogenous mycorrhizae in the root of the host plant was investigated using the carrier prepared according to the present invention. First, seeds of sorghum and Sudanese grass were sterilized with 70% ethanol and 2% NaOCl, and then thoroughly washed with sterilized water. The sterilized seeds were sown on a culture medium for plant cell culture, and the seeds were transferred to a 12-well plate for primary seeding. Seed germination and primary growth were carried out in aseptic tissue culture room and maintained at 25 ℃.

The young seedlings with the leaves from the 12-well plate were transplanted into the first small port. The transplanting soil was a mixture of soil, vermiculite and perlite in a ratio of 5: 1: 1. A seedling having a uniform growth was selected from seedlings that had been activated in a small port and four kinds of specific pores and particles (hereinafter referred to as " pore-controlled biochar ") were added to bio- And then fixed for 1 week after implantation.

The soil inoculated with the endogenous mycorrhizal fungus, Glomus sp., Was finely ground and transplanted into the pore-regulated biochar, grown for one month, and naturally infected with glomus species.

Comparative Example 1

The same procedure as in Example 1 was carried out except that the carrier in which the soil and the soil were mixed in a ratio of 1: 1 was used instead of the void-controlled biochar of Example 1.

Comparative Example 2

The experiment was carried out in the same manner as in Example 1, except that the biochar charcoal itself, that is, the uncooled biochar charcoal was used instead of the pore-regulated biochar of Example 1.

Comparative Example 3

The experiment was conducted in the same manner as in Example 1 except that activated carbon was used instead of the void-regulated biochar of Example 1.

Comparative Example 4

The experiment was carried out in the same manner as in Example 1, except that the zeolite was used in place of the void-regulated biochar of Example 1.

Comparative Example 5

The experiment was carried out in the same manner as in Example 1 except that charcoal was used instead of the void-regulated biochar of Example 1.

Infection rate survey

One month after the inoculation of the glomus species, the crops of each carrier were collected and the root slices were obtained through random sampling for each example and each comparative example. The infection rate of glomus species was investigated through staining and microscopic observation of the obtained root slices. Survey items include hyphae infection rate in plant roots, spore infection rate in plant roots, and g per gallon per carrier.

Experiment result

The infection rate of spore and mycelia of endemic mycorrhizae against one of the host plants was investigated according to the above procedure, and the results are shown in Table 2.

Crop Spore spawn Infection rate (%) ratio(%) Infection rate (%) ratio(%) Example 1 10.53 231.43 21.62 146.38 Comparative Example 1 4.55 100 14.77 100 Comparative Example 2 3.77 82.86 16.98 114.96 Comparative Example 3 6.31 138.68 15.79 106.91 Comparative Example 4 5.45 119.78 16.36 110.77 Comparative Example 5 3.80 83.52 13.92 94.25

The infection rate of other carriers was compared and analyzed based on Comparative Example 1 in which a common soil was used as a carrier component, that is, the infection rate of Comparative Example 1 was 100%.

In Example 1 of the present invention using pore-regulated biochar, the spore was increased by about 131% compared to Comparative Example 1 using a common soil as a carrier component, and about 148% was found in Comparative Example 2 using general biochar, And increased by about 97% as compared with Comparative Example 3 using activated carbon. In addition, it was confirmed that the spore infection rate of Example 1 according to the present invention was increased by about 111% even for Comparative Example 4 using zeolite, and also for Comparative Example 5 using charcoal.

In Example 1 according to the present invention, the infection rate of mycelia was increased by about 46% as compared with that of Comparative Example 1, and about 31%, 39%, and 30%, respectively, in order of Comparative Example 2, About 35% and about 52%, respectively.

Next, the spore and mycelial infection rates of endogenous mycorrhizae against one of the host plants, Sudan grass, were investigated according to the above procedure, and the results are shown in Table 3.

Plant name: Sudanese Grass Spore spawn Infection rate (%) ratio(%) Infection rate (%) ratio(%) Example 1 13.10 113.03 27.27 113.58 Comparative Example 1 11.59 100 24.01 100 Comparative Example 2 12.20 105.26 24.39 101.58 Comparative Example 3 11.11 95.86 24.07 100.25 Comparative Example 4 10.13 87.40 25.32 105.46 Comparative Example 5 9.23 79.64 23.08 96.13

The infection rate of other carriers was comparatively analyzed on the basis of Comparative Example 1 in which a common soil was used as a carrier component, that is, the infection rate of Comparative Example 1 was 100%.

In Example 1 of the present invention using pore-regulated biochar, the spore was increased by about 13% as compared with Comparative Example 1 using a common soil as a carrier component, and about 7% as compared with Comparative Example 2 using general biochar, And it was increased by about 17% as compared with Comparative Example 3 using activated carbon. It was also confirmed that the spore infection rate of Example 1 according to the present invention was increased compared to Comparative Example 4 using zeolite and Comparative Example 5 using charcoal.

In addition, in Example 1 according to the present invention, the infection rate of mycelia was increased by about 13% as compared with Comparative Example 1, and it was confirmed that the infection rate was increased more than Comparative Example 2, Comparative Example 3, Comparative Example 4 and Comparative Example 5.

Finally, the number of spores per gram of carrier for each crop was examined, and the results are shown in Table 4.

Sorghum Sudanese Grass Porosity
(ea / g) ratio(%) Porosity
(ea / g) ratio(%) Example 1 36.67 113.42 36.33 119.78 Comparative Example 1 32.33 100 30.33 100 Comparative Example 2 20.67 63.93 23.33 76.92 Comparative Example 3 34.00 105.17 35.67 117.61 Comparative Example 4 24.67 76.31 27.67 91.23 Comparative Example 5 31.67 97.96 35.33 116.49

Based on Comparative Example 1 using a common soil as a carrier component, that is, the percentage of the spore of the other carrier was compared with 100% of the spore of the comparative example 1.

As shown in Table 4, the number of spores per unit gram of carrier was found to be larger than that of Example 1 according to the present invention.

The spore infection rates and mycelial infection rates of the respective carriers were higher than those of the Sudanese grams, and the infection rate and the number of spores per gram of each of the comparative examples of Example 1 were higher than those of the case of Example 1 In the case of carrier with controlled biochar, the infection rate and the number of spores were increased to a level more significant than other carriers.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments. It is clear that the present invention can be suitably modified and applied in the same manner. Therefore, the above description does not limit the scope of the present invention, which is defined by the limitations of the following claims.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the scope of the present invention.

Claims (6)

A biomass preparation step (S10) of preparing biomass by drying after removing impurities by crushing and screening biomass;
(S20) a step of preparing bio-char to prepare bio-char by thermally decomposing the biomass by indirectly heating the prepared biomass in an air-free atmosphere in a sealed container;
The produced biochars are sieved while being sieved, the first particles having a particle size of 2.0 mm to 3.0 mm, the second particles having a particle size larger than 0.5 mm and smaller than 2.0 mm, and a second particle having a particle size larger than 0.05 mm and 0.5 mm (S30) of dividing the particles into four particle sizes of a third particle smaller than the first particle and a fourth particle smaller than 0.05 mm;
By volume of the first particles, 50 to 60 volume% of the first particles, 20 to 30 volume% of the second particles, 10 to 20 volume% of the third particles, and 10 volume% A particle mixing step (S40) of mixing four particles;
The method of claim 1,
The method according to claim 1,
In the biomass preparation step (S10), the biomass is dried to a moisture content of 20% or less.
The method according to claim 1,
Wherein the heating temperature of the biomass is 350 ° C. to 450 ° C. in the step (S 20) of producing the biochar.
The method according to claim 1,
In the particle sorting step (S30), the size of the pores of the first particles is larger than 50 mu m, the size of pores of the second particles is 10 mu m to 50 mu m, the size of pores of the third particles is 5 mu m And the size of the voids of the fourth particles is smaller than 5 占 퐉.
The method according to claim 1,
In the biomass preparation step (S10), the biomass is a woody biomass.
An endogenous mycaminocyte carrier produced by the method for producing an endogenous mycorrhizal carrier according to claim 1.

Images