WO2012008634A1 - Method for mass-producing lichen thalli, method for restoring ecology using thalli produced thereby, and composition for restoring ecology - Google Patents

Abstract

The present invention relates to: a method for restoring the ecology of wastelands such as deserts, abandoned coal mines, oil-polluted areas, volcano-affected areas, areas in which harmful minerals or industrial wastes were discarded, rocky lands, and bare mountains, by using algae with which lichens have a symbiotic relationship, cyanobacteria, lichen-forming fungi, or thalli; and a composition for restoring ecology used for the method. Also, the present invention relates to a method for mass-producing thalli needed in restoring the ecology of the wastelands. The present invention can prevent desertification and yellow dust storms, and reform polluted areas such as abandoned coal mines and oil-polluted areas to be more conducive to the growth of plants, thereby preventing ecological and environmental pollution from deserts, abandoned coal mines, and oil-polluted areas.

Description

[Revision according to Rule 26.05.10.2010] Mass production method of lichen lichen body, ecological restoration method using limb body produced by it, and composition for ecological restoration

The present invention is a method for restoring the ecology of the wasteland such as desert, coal mines, oil land, etc. using lichens, an ecological restoration composition therefor, and a method for mass production of lichens of lichens required for the ecological restoration method and composition. It is about.

In general, desert areas are barren lands with little precipitation and large vegetation. Currently occupying more than one third of the land on earth, these desert areas are gradually expanding.

The main cause of such desertification is considered to be a combination of artificial and climatic causes. Artificial causes include irrigation, deforestation and environmental pollution. Climate causes include drought and drying. In particular, from the climatic point of view, the area where precipitation is less than the possible evaporation amount is a dry land, and desertification is increasing rapidly around such dry land. According to UNEP's 1984 report, deserts are expanding at an annual rate of 60,000 km ² to the surrounding areas of existing deserts and beyond. As a result, countries suffering severe drought or desertification have joined together to form a desertification agreement.

The desert is a dry area where the temperature difference between day and night is severe and there is not enough rain for people or animals and plants. The water is supplied to these areas by converting seawater to freshwater or groundwater. The methods of developing and using are being used. However, these methods have a problem in that it takes a huge cost and time to obtain water because the water supply device must be provided separately in the area far from the sea, and the excavation is required to dig quite deep for the development of groundwater. In other words, the existing irrigation system for desert greening has the trouble of supplying water to planting trees through the watering facilities every day or every 3 to 5 days, and more than 65% of the water supply evaporates during the water supply or the root of the plant It was lost to the place where it was not reached, and the huge waste of water was repeated. However, where there is no such water irrigation system or where irrigation facilities cannot be installed, planting is not possible, or there is a restriction to plant only trees that can tolerate drought well. However, this also has a very high mortality. In addition, the desert is very sandy because of the significant temperature difference between day and night. Although the sand winds plant the desert, there is a problem of being buried in sand before it grows properly.

On the other hand, soils contaminated by oil spills for many years or decades in storage facilities such as large oil storage facilities, oil pipelines, and gas stations stop the growth of plants such as soil crops and cause secondary pollution such as groundwater pollution. In addition, the abandoned coal mines left after digging coal dust and coal from coal mines have very low acidity and are covered with black ore, so the soil moisture is very low and absorbs a lot of heat in the summer to maintain a very high surface temperature. Like oil-polluted soils, plants such as soil crops stop growing and cause secondary pollution such as groundwater pollution.

As the environmental damage to soil ecosystem and human body caused by soil pollution is highlighted, various soil restoration methods have been developed and applied in developed countries such as the United States. These methods can be divided into biological restoration methods and physical and chemical restoration methods depending on the characteristics of the technology applied. However, these methods have been limited depending on the cause of the soil pollution, and it is inevitable to have a cost when restoring the contaminated soil over a certain size.

Therefore, the inventors of the present invention overcome the technical limitations that must be applied according to the regional characteristics of the desert area, contaminated soil area, etc., respectively, and prevent desertification without expensive irrigation facilities, and regardless of the cause of pollution As a result of an effort to develop an ecological restoration method of barren land that can be applied to restore the contaminated soil, the present invention has been completed.

One object of the present invention is to use lichen symbiotic algae, cyanobacteria, lichen-forming molds, or lichen bodies to make deserts, coal waste mines, oil pollutants, volcanic ash, hazardous mineral or industrial waste sites, rocky areas, It is to provide a way to restore the ecology of the barren mountains, such as bald mountain.

It is another object of the present invention to provide a composition for restoring ecological wasteland for restoring the ecology of the wasteland.

Another object of the present invention is to provide a method for producing a large amount of limbs required for restoring the ecology of the barren.

In one embodiment, the present invention provides a method for restoring the ecology of barren land using lichen symbiotic algae, cyanobacteria, lichen forming fungi, or lichen bodies.

In the present invention, "lichen lichen" refers to a symbiotic complex of lichen-forming fungi and algae and / or cyanobacteria.

In the present invention, “thallus” is composed of lichen symbiotic algae or cells of cyanobacteria, and mycelium of lichen-forming fungi, and refers to lichen nutrients for germination, degeneration or nutrition.

In one specific embodiment, the present invention comprises the steps of: (a) immobilizing at least one of lichen symbiotic algae and cyanobacteria by mixing with an immobilization carrier; (b) introducing the immobilized carrier into a wasteland for ecological restoration; (c) inoculating the lichen forming fungus or lichen body on the introduced immobilization carrier; And (d) forming and propagating lichens and growing them into lichens.

In the wasteland ecological restoration method of the present invention, the first step is to immobilize (a) at least one of lichen symbiotic algae and cyanobacteria by mixing with an immobilization carrier.

The "lipid symbiotic algae" or "algae" refers to green algae or green algae that form part of lichens and their lichens and are capable of photosynthesis by chlorophyll. Algae included in the present invention are not limited to these, but are not limited to T. asymmetrica , T. impressa , T. jamesii , T. usneae , Magna ( T. magna ), T. erici , T. corticola , etc. Trebouxia , Pseudotrebouxia , Stichococcus diplosphaera , etc. styryl Coco carcass (Stichococcus) there is a genus, etc. Transistor tepo Liao (Trentepohlia) in such Tina (Trentepohlia abietina), O rugi labor (Trentepohlia aeruginosa), Aurea (Trentepohlia aurea), are Bohrium (Trentepohlia arborum) in the AVI .

The "cyanobacteria" refers to bacteria that make up lichens and parts of their lichens and are photosynthetic in aquatic life. Examples thereof include furnace Stock comb Sori (Nostoc commune), no stock Carne Titanium (Nostoc carneum), no stock plastic gelri formate methoxy (Nostoc flagelliforme Born et Flsh) no stock (Nostoc), A ringeu via Cri Toba group or tooth, such as ( Lyngbya crytovainatus Schk) such ringeu vias (Lyngbya), a micro collection mouse Bagi or tooth (Microcoleus Vaginatus (Vauch) Gom) including a micro-collection-house (Microcoleus), An Analog vena (Anabaena), a croissant Lactococcus epitaxial repetition of the Goose ( Chrococcus epiphyticus), including the Black Cocker tm (Chrococcus) in, in Cap-four (Gloecapsa) to him, FORT Medium Te Nuevo (Phormidium tenue), such as the Fortis Medium (Phormidium) in, ski weekends and Cinema Now Pony Qom (Scytonema japonicum), etc. Scytonema genus, Synechocystis pevalekii , etc. Synechocystis genus, Calothrix genus, Tolypothrix genus.

The immobilized carrier prevents lichen symbiotic algae and / or cyanobacteria, or lichen lichens, detailed below, from moving to areas other than the barrens of interest due to natural environmental factors, such as wind and rain, and then inoculated. It is a matrix composed of polymer material of natural polymer which maintains biological bond with lichen forming mold and enables to grow into lichen.

The natural polymers include, for example, alginic acid or salts thereof, polysaccharides such as cellulose, polyhydroxyalkenate, and the like. Preferably the material of the immobilization carrier is alginic acid or a salt thereof. More preferably, the immobilization support is made of alginic acid salt. The salt constituting the alginic acid salt is an alkaline earth metal, preferably sodium or potassium. In addition, the alginic acid salt preferably has a molecular weight of 10,000 to 1,000,000 Da.

In addition, it is preferable that the immobilized carrier maintains a water retention force of a predetermined level or more. The water retention above the predetermined level refers to the ability to retain at least 15%, preferably at least 20%, more preferably at least 22%, and most preferably at least 25% moisture after one day relative to the initial amount of water.

The polymer material of the natural polymer used in the immobilization support of the present invention is 1.0 to 3.5% by weight, preferably 1.0 to 3.0% by weight, more preferably 1.0 to 2.0% by weight, most preferably 1.2 to 1.8% by weight Mixed by content. If it is less than the above range there is a problem that the growth amount of lichen symbiotic algae or cyanobacteria included in the immobilized carrier gradually decreases over time. And if it exceeds the above range, there is a problem that the growth amount of lichen symbiotic algae or cyanobacteria included in the immobilization carrier does not increase any longer (see Example 7).

In this step, the lichen symbiotic algae and / or cyanobacteria may be further mixed with an immobilized carrier, followed by extrusion and molding in an extruder to produce a molding. The shape of the molding can be molded into various shapes such as circular or polygonal by appropriately selecting an extrusion die. The diameter of the extrusion die is suitably 1 to 20 mm in diameter or length between opposite sides. This takes into account the barren environment in which lichens, lichen lichens, lichen-forming molds, algae, and cyanobacteria will grow, for example, if they are desert, they should be large enough to not be lost in the desert sand. The aperture size may also be appropriately selected depending on the size of the treatment facility.

Next, (b) introducing the immobilized carrier into the wasteland for the purpose of ecological restoration. That is, the step of introducing and fixing the lichen symbiotic algae or cyanobacteria immobilized by the immobilization carrier by spraying or the like on the barren for the purpose of ecological restoration.

Waste land or wild land for the purpose of ecological restoration includes coal mines, oil pollution, deserts, volcanic ash, places where hazardous minerals or industrial wastes are disposed, rocky land, bald mountain, It refers to places where spores, eggplants, flowering plants, monocots and dicots cannot grow and reproduce. The nematodes and the plants can be grown and reproduced only when the environment suitable for growth is provided. The environment suitable for growth will vary slightly from plant to plant, but will generally be light, growth temperature, water in the soil, nutrients and pH. The barren, however, is inadequately configured. For example, the nutrients that plants absorb from the soil are essential for plant growth. The essential elements are carbon (C), hydrogen (H), oxygen (O), nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg) and sulfur (S) It is composed of elements and trace elements of iron (Fe), manganese (Mn), copper (Cu), zinc (Zn), boron (B), molybdem (Mo), and chlorine (Cl). However, the barren is a very small element or element necessary for the growth of the plant, or the presence of an excessive or elemental element (s) that makes it difficult to grow the plant, such as heavy metals, it is impossible to settle the plant. Plants also absorb water from the soil and use it for growth. Barrens, such as deserts, lack such water and are very difficult to grow. In addition, plants can grow in neutral or weakly acidic to weak alkalinity. In the case of coal mines or oil papers, the content of heavy metals is very high and acidic properties make it impossible to grow plants. Therefore, ecological restoration is to restore such wasteland to the soil where plants can grow to prevent environmental and ecological pollution caused by wasteland. Environmental and ecological pollutions include, for example, in vivo pollution due to yellow dust generation by deserts, soil and groundwater contamination by oil and coal waste mines, landslides caused by cracks in rock, and the like.

In the present invention, the introduction refers to the arrangement of the lichen symbiotic algae and / or cyanobacteria is immobilized to the surface or the surface of the barrens to a depth of 1 cm or less. Since the immobilized carrier has a size of 1 to 20 mm, there is no fear of loss due to wind and rain, and thus, it is possible to obtain a sufficient soil improvement effect for the ecological restoration just by placing it on the surface of the wasteland. In addition, even if the immobilized carrier is placed at a depth of 1 cm or less from the surface of the wasteland, such arrangement is possible because air and light necessary for growing lichens can be obtained from the natural environment by the voids between soils. However, when the immobilized carrier is disposed at a depth exceeding 1 cm, sufficient air and light are not available to lichen symbiotic algae and cyanobacteria, and thus lichen lichen body formation and lichen growth are slow, which is not preferable. Here, the arrangement is preferably such that the ratio of the immobilized carrier to the barren is 20% or more, it can be made by a method such as spraying or planting the immobilized carrier.

Next, (c) inoculating lichen forming fungus or lichen body on the immobilized carrier, and (d) forming and propagating lichen body and growing into lichen.

The lichen-forming mold refers to fungus that form part of lichens and form a symbiotic relationship with algae or cyanobacteria as described above. The lichen-forming fungus can be any one that can form a symbiotic relationship with algae or cyanobacteria, but is not limited thereto.Aspicilia) Species, Caloplaca (Caloplaca), Cladonia (Cladonia) Species, Coleema tenas (Collema tenaxCholema (C, etc.)ollema) Species, dematocarpon (Dematocarpon), Dipronistus (Diploschistes) Chong, Endocapon FussilEndocarpon pusillumEndo carphone (E) such asndocarpon) Species, Plugencia (Flugensia) Species, graffiti (Graphis) Bell,Griporecia (Glypholecia) Bell, GiroforaGyrophora) Species, heterodermia (Heterodermia) Species, fishia (Physica) Species, LecanooraLecanora) Species, PeopiaPhaeophyscia) Species, Pisconia (Physconia) Bell, PsoraPsora) Species, Parmeria (Parmelia) Bell, RaizopuraRhizoplaca) Bell, Lamarina (Ramalina) Bell, stereo cowlon (Stereocaulon) Species, Spaeroporus (Sphaerophorus) Species, Teroshiciss (Teloschistes) Species, Tonyia (Toninia) Species, UmpiricariaUmbilicaria) Species, Usnia (Usnea) Species, PhilophorusPilophorus), Xantho Palmellia (Xanthoparmelia) Species, Zantharia (Xanthoria) Species.

The lichen is composed of lichen symbiotic algae or cyanobacteria cells, and the mycelia of lichen-forming fungi, as described above, refers to lichens for lichen, fever or nutrition. Since these limbs are very small in their natural state, it is preferable to use limbs produced by the mass production method of lichen limbs, which will be described later in the present invention. Inoculation of the lichen body has the advantage of shortening the formation and breeding time of the lichen body than inoculation of the lichen-forming mold.

Said inoculation refers to the denting of lichen-forming molds or lichens on algae or cyanobacteria. This inoculation is achieved by flocculation on algae or cyanobacteria with 1 to 20 cells per cm ² , preferably 2 to 12 cells per cm ² , more preferably 4 to 8 cells per cm ² for lichen forming fungi. It is preferable that the tooth lichen-forming mold to be toothed has a diameter of 2 mm or less, more preferably 1 to 2 mm per cell. In the case of lichens, it consists of flocculation on algae or cyanobacteria in 1 to 20 individuals per cm ² , preferably 2 to 12 individuals per cm ² , more preferably 4 to 8 individuals per cm ² . Magazine body to the tooth shape preferably has a surface area per object 4 mm ² or less, preferably of 2 to 4mm ^2.

A part of algae or cyanobacteria immobilized on the carrier by the inoculation and a part or lichen body of lichen-forming molds are biologically combined to form lichens of lichens, and grow to lichens by breeding lichens over time. Done.

Steps (b) and (c) of the present invention may reverse the order. Specifically, after (c) attaching lichen-forming mold to the immobilized carrier to form lichens and growing them into lichens, (b) introducing the immobilized carrier into wasteland for ecological restoration. have.

Even in this alternating order, lichen symbiotic algae and / or cyanobacteria and lichen forming fungi in the immobilized carrier are capable of biological bonding, from which lichen lichens form and multiply and grow into lichens.

On the other hand, in the order of step (b) and (c), after step (c), or in the order of step (c) and (b), step (b) is introduced into the barren for the purpose of ecological restoration. Applying the coagulant to the immobilized carrier to a thickness of at least half the diameter of the immobilized carrier, preferably from 0.5 to 2 times the diameter of the immobilized carrier.

The coagulant is for preventing the immobilized carrier from moving out of the wasteland for ecological restoration by the natural environment such as wind or rain. In addition, the use of the coagulant increases the compressive strength and water retention of the biological soil cluster (biological soil clust) to be formed on the barren in the future than the use of only the immobilized carrier, so that the ecological restoration of the wasteland soil is easier have.

The coagulant may be any one that can achieve the above object, for example, polylactide, polyglycolide, poly (lactide-co-glycolide), poly-β-hydroxybutyric acid, polyan Hydrides, polyorthoesters, polyurethanes, polyamide polycarbonates, polyethylene, polypropylene, poly (ethylene glycol), poly (ethylene oxide), poly (ethylene terraphthalates), polyvinyl alcohols, polyvinyl ethers, polyvinyl esters , Poly (vinylchloride), polyvinylpyrrolidone, polysiloxane, poly (vinyl alcohol), poly (vinyl acetate), polystyrene, polyurethane and copolymers thereof, derived cellulose such as alkyl cellulose, hydroalkyl cellulose, cellulose ether , Cellulose ester, nitrocellulose, methyl cellulose, ethyl cellulose, hydropro Synthetic cellulose such as cellulose hydroxy, hydroxy-propyl methyl cellulose, hydroxybutyl methyl cellulose, cellulose acetate, cellulose propinate, cellulose acetate butylate, cellulose acetate phthalate, carboxyethyl cellulose, cellulose triacetate and cellulose cellulose sodium salt, Polymers of acrylic acid, derivative copolymers thereof including esters or methacrylates, poly (methyl methacrylate), poly (ethyl methacrylate), poly (butyl methacrylate), poly (isobutyl methacrylate) , Poly (hexyl methacrylate), poly (isodecyl methacrylate), poly (isopropyl acrylate), poly (isobutyl acrylate) and poly (octadecyl acrylate), poly (butyl acid), poly (Valeric acid) and poly (lactide-co-caprolactam), these Copolymers and mixtures thereof.

On the other hand, in the method of ecological restoration of the barren of the present invention, the lichen forming mold or lichen body can be immobilized by mixing with one or more of lichen symbiotic algae and cyanobacteria from the step (a). In this case, there is an advantage in that the lichen forming fungus or lichen body can be omitted on the immobilized carrier including any one or more of lichen symbiotic algae and cyanobacteria.

Accordingly, the present invention provides another specific embodiment, comprising: (a) immobilizing at least one of lichen symbiotic algae and cyanobacteria, lichen forming mold or lichen by mixing with an immobilizing carrier; (b) introducing the immobilized carrier into a wasteland for ecological restoration; (c) forming and propagating lichen bodies from the immobilized carriers and growing them into lichens.

By the above-described method of ecological restoration of wasteland, biological soil clusters by lichens are formed on the wasteland for the purpose of ecological restoration, thereby inducing a primary transition.

The biological soil cluster is a microbial mass including several to several tens of centimeters of lichens formed on the soil surface of the barren soil, which is almost the first of the natural transition process. In general, the process of changing the barren soil into soil where plants can grow is called cloth. In the transition process, lichens invade, form clusters, and organic matters accumulate in the soil. Then, the accumulated organic matter is used as nutrients to settle lichens, and then annual herbaceous plants, perennial herbaceous plants, amniotic forests. In this order, the positive and negative forests are formed. These natural transitions require tens to hundreds of years, especially because lichens naturally invade, settle, and form clusters. However, when using the wasteland ecological restoration method of the present invention, since the settlement of lichens and the formation of clusters are made in a short time, the natural transition process can be made in a short time. That is, lichen symbiotic algae, cyanobacteria, and lichen forming fungi used in the ecological restoration method of the wasteland of the present invention are natural environments such as rain and wind that interfere with the growth of lichens and lichens and the formation of biological soil clusters on their immobilized carriers. It grows in a short period of 6 months to 1 year or 2 years without affecting to form biological soil clusters. After the formation of the biological soil clusters, the properties of the barren soils are not only changed to be suitable for the growth of nematodes and plants such as moss, but, for example, deserts are prevented from blowing sand by desert winds. have.

As another aspect, the present invention provides a composition for the ecological restoration of barrens to be used in the method of ecological restoration of the barrens. Specifically, the present invention provides a composition for ecological restoration of wasteland comprising at least one of lichen symbiotic algae and cyanobacteria, lichen forming mold or lichen body, and an immobilized carrier.

Since any one or more of lichen symbiotic algae and cyanobacteria, lichen-forming mold or lichen body, and immobilized carrier included in the wasteland ecological restoration composition of the present invention are as described above, a detailed description thereof will be omitted.

The lichen is settled and grows in the wasteland by the composition of the wasteland ecological restoration of the present invention. Lichens are also called early intruders, because lichens can settle on a rock that has emerged as well as a plant-like form on a newly emerged surface for the first time. Lichen can also act as an early invader because of its ability to withstand droughts over long periods of time. Lichens can obtain the nutrients necessary for growth from the atmosphere, absorb anything dissolved in moisture in the atmosphere, and obtain carbohydrates, photosynthetic products from symbiotic algae in the body. And lichens have very small propagules and can settle anywhere except on extremely flat surfaces. Many lichens have other advantages as early invaders. If they have symbiotic algae, or have a cephalodia, a peculiar structure in which the algae inhabit, these symbiotic cyanobacteria can use the nitrogen they have fixed. Therefore, even in a nitrogen-deprived environment, such as a newly emerged rock surface, these lichens can be supplied with nitrogen and carbon on their own, making them an extremely favorable location for early settlement. And unlike in rocks, lichens in soil are absolutely inferior in competition with fast-growing creatures such as herbaceous plants, moss, and weeds. However, lichens occupy an absolute growth position in vast deserts and polar regions that are not suitable for herbaceous plants, moss, weeds, etc.

Therefore, the second transition of lichens, weeds and trees after the settlement of lichens has been developed when developing methods to artificially settle lichens in extreme environments where herbaceous plants, moss, and weeds are inferior as early intruders. It is suggested by the present inventors that the present invention can lead to extreme soil environmental changes and ecosystem changes. In one specific implementation, it can be seen that lichens in the cladonia collected from coal abandoned mines are settled and grown in coal abandoned mines, and that lichens and plants grow only in the lichen settlements (see FIG. 1). In addition, the analysis of liposomes in lichens living in coal mines shows that lichens are resistant to heavy metals, so lichens can inhabit as an early intruder even in extreme environments where vegetation such as abandoned mines cannot grow. (See Table 1).

In the present invention, the lichens are settled in rocky land where moss and plants cannot grow, deserts, polluted lands such as coal and oil, volcanic land, places where hazardous minerals or industrial wastes are disposed, rocky land, bald mountain, etc. Includes all lichens that can grow. For example, but are not limited to, but ah RY Syria (Aspicilia), A knife Plaka (Caloplaca), A Cloud Macedonia (Cladonia), A Collet Do (C ollema) in, for Mato carboxylic phone (Dematocarpon), A deep The genus Diploschistes , genus E ndocarpon , genus Flugensia , genus Glypholecia , genus Gyrophora , genus Heterodermia , genus pisia ( Physica ), Lecanora , Phaeophyscia , P hysconia , Psora , Parmelia , Rhizoplaca , La Ramalina genus, Stereocaulon genus, Sphaerophorus genus, Teloschistes genus, Toninia genus, Umbilicaria genus, pilopo Pilophorus genus, Xanthoparmelia genus, Xanthoria ) Lichen limbs.

Among the various lichen genus that can be settled and grown in areas where moss and plants cannot grow, a lichen suitable for the purpose of ecological restoration is selected from the lichen symbiotic algae or cyanobacteria, lichen bodies, and Lichen forming mold can be isolated and cultured. In particular, the limb can be produced by the mass production method of the limb as described below. In one specific implementation, lichens suitable for ecological restoration of coal abandoned mines are lichens in the genus Cladonia. The lichens of the genus Cladonia preferably include Cladonia macilenta , Cladonia humilis , Cladonia ramulosa , and the like.

In one specific implementation, the soil pH was higher in LCC soils than in NBC soils, as compared to lichen-colonized coalmine (LCC) and noncolonized bare coalmine (NCC). Ca and Mg, which were not detected at all, were detected in the LCC soil, and the heavy metal concentrations of Fe, Cu, Ni, and Mn were significantly reduced in the LCC soil compared to the NBC soil. These results indicate that soils in lichen habitats can transform barren traits into an environment suitable for plant growth.

In another specific practice, bacteria and fungi in LCC soils were analyzed by analyzing the population of microorganisms in lichen-colonized coalmine (LCC) and noncolonized bare coalmine (NCC). It can be seen that the number of individuals is significantly higher. In addition, as a result of examining the enzymatic activity and metabolic activity of the microorganisms in these soils, it can be seen that the amount of enzyme activity and metabolic activity increased in the LCC soil than in the NBC soil. More specifically, cellulase, beta-glucosidase, urease and invertase activity, which showed extremely low activity in NBC soils, increased significantly in LCC soils. It was. When the hydrothermal extract of lichen was added to each soil, the metabolic activity of microorganisms was increased in all soils. In particular, LCC soils increased only 2.3 times, while 8.2 times in NBC soils. These facts suggest that lichen habitat stimulates and increases the activity of microorganisms in barren land.

In another specific embodiment, when only lichens, algae or cyanobacteria were treated to soil alone, they did not form biological soil clusters, whereas the bioremediation soil modifying agent of the present invention was confirmed to form biological soil clusters. Can be. In addition, the formed cluster has a constant thickness, exhibited a certain level of compressive strength, it can be confirmed that the subsequent transition is possible by maintaining a moisture holding capacity of a certain level or more.

In another aspect, the present invention provides a method for mass production of lichens of lichens. Specifically, the present invention comprises the steps of artificially culturing lichen symbiotic algae or cyanobacteria; (b) inoculating the lichen forming fungus on the alga or cyanobacteria to form lichens; And (c) inducing the propagation of lichens by placing the algae or cyanobacteria on which the lichens are formed in a material according to the growth environment of the algae or cyanobacteria.

In the present invention, "lichen lichen" refers to a symbiotic complex of lichen-forming fungi and algae and / or cyanobacteria.

In the present invention, “thallus” is composed of lichen symbiotic algae or cells of cyanobacteria, and mycelium of lichen-forming fungi, and refers to lichen nutrients for germination, degeneration or nutrition.

In the lipoprotein mass production method of the present invention, the first step is artificially culturing lichen symbiotic algae or cyanobacteria.

In the present invention, "lipoid symbiotic algae" or "algae" refers to green algae or cyanobacteria that form part of lichens and their lichens and are capable of photosynthesis by chlorophyll. Algae included in the present invention are not limited to these, but are not limited to T. asymmetrica , T. impressa , T. jamesii , T. usneae , Magna ( T. magna ), T. erici , T. corticola , etc. Trebouxia genus, Pseudotrebouxia genus, Stichococcus diplosphaera , etc. styryl Coco carcass (Stichococcus) there is a genus, etc. Transistor tepo Liao (Trentepohlia) in such Tina (Trentepohlia abietina), ah rugi labor (Trentepohlia aeruginosa), Aurea (Trentepohlia aurea), are Bohrium (Trentepohlia arborum) in the AVI .

In the present invention, "cyanobacteria" refers to bacteria that make up lichens and a part of their lichens, which are photosynthetic of aquatic life. Examples include Nostoc commune , Nostoc carneum , and Nostoc genus Nostoc flagelliforme Born et Flsh, Lyngbya Lygbya genus, such as crytovainatus Schk, Microcoleus genus, such as Microcoleus Vaginatus (Vauch) Gom, Anabaena genus, Crocoker tm Epipitigus ( including Chrococcus epiphyticus), including the Black Caucus (Chrococcus) in, CAP Corporation (Gloecapsa), a PORT Medium Te Nuevo (Phormidium tenue) FORT Medium (Phormidium) in, ski weekends and Cinema Now Pony Qom (Scytonema japonicum) in such a thereof Scytonema genus, Synechocystis pevalekii and the like Synechocystis genus, Calothrix genus, Tolypothrix genus.

The cultivation of lichen symbiotic algae or cyanobacteria can be cultured by methods well known in the art depending on the kind of lichen symbiotic algae or cyanobacteria to be cultured. For example, using a BBM culture medium for green algae and BG-11 culture medium for cyanobacteria, shaking incubator, agitating incubator or bottom air support at 15 to 25 ° C under light conditions (light intensity 100-400 PAR) for 12 hours. It is preferable to culture | cultivate, supplying artificial air using the apparatus of a group.

In the mass production method of lichen body according to the present invention, the next step is to inoculate lichen-forming mold to algae or cyanobacteria cultured in the first step to form lichen body.

In the present invention, “lichen formin fungi” refers to fungus that form part of the lichen and form a symbiotic relationship with algae or cyanobacteria. Lichen-forming molds included in the present invention can be any form as long as they can form a symbiotic relationship with algae or cyanobacteria, but are not limited thereto.Aspicilia) Species, Caloplaca (Caloplaca), Cladonia (Cladonia) Species, Coleema tenas (Collema tenaxCholema (C, etc.)ollema) Species, dematocarpon (Dematocarpon), Dipronistus (Diploschistes) Chong, Endocapon FussilEndocarpon pusillumEndo carphone (E) such asndocarpon) Species, Plugencia (Flugensia) Species, graffiti (Graphis) Bell,Griporecia (Glypholecia) Bell, GiroforaGyrophora) Species, heterodermia (Heterodermia) Species, fishia (Physica) Species, LecanooraLecanora) Species, PeopiaPhaeophyscia) Species, Pisconia (Physconia) Bell, PsoraPsora) Species, Parmeria (Parmelia) Bell, RaizopuraRhizoplaca) Bell, Lamarina (Ramalina) Bell, stereo cowlon (Stereocaulon) Species, Spaeroporus (Sphaerophorus) Species, Teroshiciss (Teloschistes) Species, Tonyia (Toninia) Species, UmpiricariaUmbilicaria) Species, Usnia (Usnea) Species, PhilophorusPilophorus), Xantho Palmellia (Xanthoparmelia) Species, Zantharia (Xanthoria) Species.

In the present invention, "inoculation" refers to the denting of lichen-forming molds to algae or cyanobacteria. Such inoculation is the lichens forming mold in such a vaccination is the case of lichens forming mold cm 1 to 20 cells per ^second, and preferably cm ² 2 to 12 cells per, more preferably cm ² birds 4 to 8 cells per or cyanobacteria It is done by healing. It is preferable that the tooth lichen-forming mold to be toothed has a diameter of 2 mm or less, more preferably 1 to 2 mm per cell.

By the inoculation, a part of algae or cyanobacteria and a part of lichen forming molds are ecologically combined, and the combination of lichens is formed. In addition, a large amount of the limb is formed by the method of germination, heat germination or nutrition propagation.

Next, the algae or cyanobacteria in which the liposomes are formed are placed on a material according to the growth environment of the algae or cyanobacteria to induce the propagation of the lichens.

When lichens are inoculated with lichen-forming molds, lichens are formed and placed in the ground according to the growth environment of algae or cyanobacteria. Positioning on the substrate according to the growth environment is, for example, algae or cyanobacteria, which preferably grow in trees, are located in the trees, and algae or cyanobacteria, which preferably grow in stones or rocks, are located in the rocks or rocks. It means to be located in the optimum material that grows according to the species of algae or cyanobacteria, such as. Therefore, the present invention according to the growth environment of algae or cyanobacteria may be specified according to the species of the algae or cyanobacteria, but is not limited thereto, for example, trees, stones, rocks, cork, sand, etc. Can be mentioned.

The algae or cyanobacteria are placed in the product according to the growth environment, and then the water and nutrients are continuously supplied according to the growth conditions of the algae or cyanobacteria to induce the propagation of the lichens.

In addition to forming lichen bodies only where the conventional lichen-forming mold was inoculated by the lichen body mass production method of lichens according to the present invention, lichens are multiplyed and lichen bodies are formed in a large amount so that lichen bodies can be used in large quantities. do. In one specific embodiment, the inventors have confirmed that the lichen body can be mass produced by the lichen body mass production method of lichen limb of the present invention (see Example 6).

According to the present invention, it is possible to restore ecology to a soil in which plants can grow in the wasteland where plants such as deserts, abandoned mines, oil pollutants, and the like cannot grow. In particular, by this ecological restoration it will be possible to prevent the global warming and environmental and ecological destruction due to the prevention of environmental pollution by the increase of the desert and the desert yellow sand, environmental pollution by waste mines and oil pollution sites. In addition, since the lichen limbs required for ecological restoration can be produced and supplied in large quantities, ecological restoration by the ecological restoration method can be effectively performed.

1 is a picture of coal mines. Most coal mines are exposed to waste coal without vegetation, and only a few areas are lichens spread in the form of patches, and plants such as moss and birch survive.

FIG. 2 shows a comparison of microbial metabolic activity in lichen-colonized coalmine (LCC) soils and lichen-colonized bare coalmine (NCC) soils.

FIG. 3 shows the results of cellulose degradability in lichen-colonized coalmine (LCC) soils and noncolonized bare coalmine (NBC) soils.

Figure 4 is an embodiment of the present invention observed the mass production process of the limb according to the progress of the process, and is an enlarged view of the formation of the limb.

FIG. 5 shows the amount of change in the number of cells in the immobilized carrier of cyanobacteria when the alginic acid is used as the immobilized carrier as an embodiment of the present invention.

6 and 7 illustrate the results of biological soil cluster formation of cyanobacteria dried or not dried after 6 months when alginic acid and / or PVA were used as an immobilization carrier.

FIG. 8 is a diagram comparing compressive strength of biological soil clusters formed by cyanobacteria using alginic acid and / or PVA as an immobilization carrier as an embodiment of the present invention.

9 is a diagram comparing water retention of biological soil clusters formed by cyanobacteria using alginic acid and / or PVA as an immobilization carrier as an embodiment of the present invention.

10 is a diagram comparing the biomass of cyanobacteria immobilized on the immobilization carrier of the present invention with the biomass of cyanobacteria not immobilized.

The invention will be explained in more detail through the following examples. These examples are presented by way of example only, so that the present invention may be more understood, these examples should not limit the scope of the invention.

Example 1: lichen collection

Three lichens were collected from abandoned mines in Hambaeksan, about 7 km northwest of Taebaek, Gangwon-do, Korea. Most of these abandoned mines were exposed to waste coal without vegetation, and some lichen groups were spread out as patches in these areas. In some patches, moss and birch (Betula Schmidt ) and birch ( Betulacostata ) were alive, but lichens also inhabited from the outer boundary to the inside of moss and plant growth (see FIG. 1). These inhabited lichens were collected and classified by Dr. Harada, stored in the Korean Literature Research Institute (Sunchon National University, Korea) and at the National History Museum and Institute (CBM) in Chiba, Japan. Has been replicated. The lichens collected were identified as Cladonia macilenta Hoffm., C. humilis (With) JR Laundon, and C. ramulosa (With) JR Laundon.

In addition, lichens from China's Shapotou and Inner Mongolia deserts were collected. The collected lichens were analyzed as Aspicilia , Caloplaca , Cladonia , Collema , Dematocarpon , Depro The genus Diploschistes , genus E ndocarpon , genus Flugensia , genus Glypholecia , genus Gyrophora , genus Heterodermia , genus pisia Physica genus, Lecanora genus, Phaeophyscia genus, P hysconia genus, Psora genus, Parmelia genus, Rhizoplaca genus, Lamarina ( Ramalina ), Stereocaulon , Sphaerophorus , Teloschistes , Toninia , Umbilicaria , Usnia Usnea) in, Philo Forus (Pilophorus), A janto Parr Melia (Xanthoparmelia), A janto Oh, it was identified as belonging to the lichen (Xanthoria). Next, from the collected lichens, Cllema tenax , Endocarpon pusillum , Flugensia species, Rhizoplaca species, Toninia species, Xanthopar Lichen-forming fungi of Xanthoparmelia species could be isolated, including Nosctoc commune , Nostoc carneum , Microcoleus sociatus , and Microreus vaginatus . Microcoleus vaginatus ), Lyngbya cryttovaginatus , Tolypothrix genus, Calothrix genus symbiotic algae or cyanobacteria were isolated.

Example 2: Component Analysis of Collected Lichens

Three lichens collected from the waste paper of Example 1 were washed twice with deionized distilled water and stored at room temperature for 3 days. Tarhanen et al., Modified to determine the concentration of components (K, Ca, Mg, Fe, Cu, Mn, Ni, As and Cr) in the lichens of stored lichens [Tarhanen, S., S. Mets, T. Holopainen and J. Oksanen. Membrane permeability response of lichen Bryoria fluscescens to wet deposited heavy metal sand acid rain. Environ. Pollut. 104 : 121-129.] Was used. Specifically, subsamples of each lichen species (500 mg) were dried at 90 ° C. for 24 hours, wet decomposition with HNO ₃ and HClO ₄ superpur (10: 3 ml) at 225 ° C. in a digestion system and a solid residue. Was isolated by filtration. Thereafter, the concentrations of the basic components were analyzed using an inductively coupled plasma spectrometer (Model D-Time 3000DC, shimadzu, Japan). The results are shown in Table 1 below, and these analyzed concentrations are expressed in μg g ⁻¹ DW.

Table 1

The results are mean and standard deviation for eight replicates for each lichen.

*: Significant differences between lichen species (P <0.001)

N.S: No significance

As can be seen in Table 1, it was found that the concentration of nutrients and heavy metals in cladonia ramurosa and cladonia humilis slightly higher than cladonia masenta. Therefore, the lichen species living in coal mines were found to be resistant to the heavy metals accumulated in their habitats.

Example 3: Comparison of Characteristics of Lichen Habitat Soil

The difference in soil properties between Lichen-colonized coalmine (LCC) soils and noncolonized bare coalmine (NCC) soils in the coal waste mine of Example 1 was analyzed. Specifically, five soil samples each from LCC soil and NBC soil were randomly collected 3 cm deep from the surface. LCC soil samples were collected from lichen carpets inhabiting abandoned mines. The collected soil was filtered through a 2 mm sieve and stored at 4 ° C. for the next analysis. For these soils, the soil pH was measured using standard laboratory techniques [Kim, H. 1995. Soil Sampling, Preparation, and Analysis . Marcel Dekker Inc., New York. In addition, to measure the heavy metals in the soil, the soil was ground with a mortar and filtered through a 100 mesh sieve. 10 g of the filtered soil was extracted with 30 ml of 0.1 N HCl at 30 ° C. for 30 minutes and filtered, and then metal concentration was measured using an inductively coupled plasma spectrometer (Model D-Time 3000DC, shimadzu, Japan).

TABLE 2

The results are mean and standard deviation of 10 samples for each soil.

**: significant difference is P <0.01

***: significant difference is P <0.001

N.D: not detected

As can be seen in Table 2 above, soil pH was higher in LCC soils than in NBC soils. LCC and NBC soils showed significant differences in some components. Specifically, the concentrations of K, Ca and Mg in LCC soils were much higher than in NBC soils. In particular, Ca and Mg were not detected in NBC soils. Na was very high in NBC soils. Heavy metal concentrations of Fe, Cu, Ni and Mn were much higher in NBC soil than in LCC soil. In contrast, there was no significant difference in As and Cr concentrations between the LCC and NBC soils.

Example 3: Population Analysis of Microorganisms on Lichen Habitat Soil

To analyze the activity of microorganisms in LCC and NBC soils, the number of bacteria and fungi was first measured by direct counting method using dilution plate method in agar and potato agar. As a result, as can be seen in Table 3, it was found that bacteria and fungi were present in LCC soils much higher than NBC soils. However, the fungus has a simple structure and uses organic compounds, indicating that the fungus is present much more than the fungus of NBC soil.

TABLE 3

Example 4 Investigation of Enzyme Activity and Metabolic Activity of Microorganisms in Lichen Habitat Soil

Enzyme activity and microbial metabolic activity (also called basal soil respiration) of microorganisms involved in organic degradation in LCC and NBC soils were measured and analyzed. The specific experimental method is as follows. All these analytical results were calculated based on the dry (105 ° C.) weight of the soil. In addition, one-way-ANOVA (SPSS Version 11.0) statistics were used to characterize significant differences between LCC and NBC soils.

4-1. Measurement of phosphatase and beta-glucosidase activity

Phosphatase and beta-glucosidase activity was determined by p -nitrophenyl phosphate disodium ( p- nitrophenylphosphatedisodium; PNPP, 0.115M) or p -nitrophenyl-beta-D-glucopyranoside ( p -nitrophenyl β-D-glucopyranoside; PNG, 0.05 M) (Mansciandaro, G., B. Ceccanti and C. Garc 1994. Anaerobic digestion of straw and pig wastewater: II. Optimization of the process.Agrochimica 3: 195- 203.). These assays are based on the release and detection of p-nitrophenol (PNP). Specifically, 2 ml of 0.1 M maleic acid buffer (pH 6.5) and 1 ml of substrate were added to 1.0 g of soil sample and incubated at 37 ° C. for 90 minutes. Thereafter, the reaction was stopped by rapidly cooling at 2 ° C. for 15 minutes. Next, 0.5 M CaCl ₂ and ₂ ml of 0.5 M NaOH were added and the mixture was centrifuged at 2000 × g for 5 minutes. Trishydroxymethyl aminomethane (THAM) was used to stop the beta-glucosidase assay. The amount of PNP was measured at 398 nm spectrophotometer. The control was done in the same manner as above, but the substrate was added to the soil just after the incubation and just before the reaction was stopped. The results are shown in Table 4 below.

4-2. Xylanase Activity Measurement

To measure xylanase activity, 1.0 g of soil samples were mixed with 10 ml substrate solution (2 M acetic acid buffer, pH 5.5) in 50 ml of xylan (1.7% w / v) from oat spelts and 10 ml of 2 M acetic acid buffer. Incubated for 24 hours. Reducing sugars released during incubation reduce potassium hexacyanoferrate (III) in the alkakine solution, and the potassium hexacyanoferrate (III) is subjected to the Prussian blue reaction. Measured by colorimetry (Kandeler, E., J. Luxhøi, D. Tscherko and J. Magid. 1999. Xylanase, invertase and protease at the soil-litter interface of a loamy sand. Soil Biol. Biochem . 31: 1171- 1179.). The results are shown in Table 4 below.

4-3. Determination of Invertase Activity

To measure sucrose enzyme activity, 1.0 g (dry weight) of soil sample was incubated in 10 ml of 50 mM sucrose solution and 10 ml of 2M acetic acid buffer (pH 5.5) at 50 ° C. for 3 hours. From this the reducing sugars were measured in the same manner as described for measuring xylase activity. The results are shown in Table 4 below.

4-4. Urease Activity Measurement

Urea dehydrogenase activity was determined using urea as a substrate, treated with 4 ml of a soil sample of 1.0 g (dry weight) dmf 0.1 M phosphate buffer (pH 7.0) and then incubated at 37 ° C. for 1.5 hours. The amount of NH ₄ ⁺ released was measured using an ammonia-selective gas electrode (Model 720A, Orion Research, USA). The results are shown in Table 4 below.

4-5. Cellulase Activity Measurement

To measure cellulase activity, 2.5 g (dry weight) of soil samples were placed in a 50 ml Erlenmeyer flask, treated with 0.5 ml toluene and buffered 2% CMC (carboxymethyl cellulose) and incubated at 30 ° C. for 24 hours. (Deng, SP and MA Tabatabai. 1994. Cellulase activity of soils. Soil Biol. Biochem . 26: 1347-1354.). The supernatant was then mixed well and centrifuged three times at 15,000 g speed for 15 minutes. Next, 10 ml of the supernatant was transferred to a 50 ml plastic centrifuge tube and treated with 2 g of potassium-saturated cation exchange resin. The mixture was shaken for 30 minutes, and then the supernatant was reduced sugar by the consumption G -Nelson method (Deng, SP and MA Tabatabai. 1994. Colorimetric determination of reducing sugars in soils. Soil Biol. Biochem . 26: 473-477.) Analyze by measurement. The blue solution for the soil extract obtained in CMC-treated soil was centrifuged once by the method described above before the 710 nm colorimetry. The results are shown in Table 4 below.

4-6. Measurement of metabolic activity of microorganisms

CO ₂ measurement method for measuring the metabolic activity of microorganisms in LCC and NBC soils (Langer, U. and T. G2001. Effects of alkaline dust deposits from phosphate fertilizer production on microbial biomass and enzyme activities in grassland soils.Environ.Pollut . 112: 321-327) was used. Specifically, 25 g of LCC and NBC soils were placed in 150 ml of polypropylene beakers, which were incubated at 25 ° C. for 5 days in a sealed 1 L glass container containing 25 ml of 0.1 N NaOH. Thereafter, 5 ml of 0.5 M BaCl ₂ and a few drops of phenolphthalein indicator were added, followed by titration with 0.1 N HCl to determine the amount of carbon dioxide released. In addition, basal soil respiration was measured after inoculating the hot water extract of lichen lichen on each soil to investigate the effect of lichen lack on soil microbial respiration. Specifically, 25 g of dried lichen lichens were extracted with 200 ml of distilled water at 90 ° C. for 3 hours. Then 5 ml of the extract was thoroughly mixed into 5 g of each soil. After drying the soil for 24 hours at room temperature, the inoculated soil was incubated in the same manner as described above. These results are expressed in μg CO ₂ -C / g soil. The results are shown in FIG. 2.

4-7. result

Table 4 below shows the results of measuring enzymatic activity of microorganisms in LCC soil and NBC soil. Soil microbial enzymes are involved in the degradation of soil organic matter, which is found to be significantly activated in LCC soils. In particular, the activity of cellulase, beta-glucosidase and sucrose enzyme was very extremely low in NBC soils compared to LCC soils. Urea dehydrogenase activity catalyzing the nitrogen mineralization of soils was also negligible in NBC soils compared to LCC soils.

Table 4

Increased activity of microbial enzymes by lichens was also confirmed by measuring the metabolic activity of microorganisms (see FIG. 1). LCC soils maintained high levels of soil respiration during incubation, while NBC soils showed almost negligible soil respiration. When hydrothermal extracts of lichen lichens were added to LCC soils and NBC soils respectively, the basal soil respiration was significantly increased during incubation (see FIG. 2). So LCC soils showed a much higher level of CO2 increase than NBC soils, but induction of soil respiration was more evident in NBC soils (8.2 fold increase) than LCC soils (2.3 fold increase). These results clearly demonstrate that lichen habitat induces and stimulates the microbial enzyme activity of coal wasteland soils.

Example 5 Investigation of Cellulose Degradability of Lichen Habitat Soil

LCC soil and NBC using the cotton-strip assay (Chew, I., JP Obbard and RR Stanforth. 2001. Microbial cellulose decomposition in soils from a rifle range contaminated with heavy metals.Environ.Pollut. 111 : 367-375.) The degradation of cellulose in the soil was measured. The original assays International Biological Program (International Biological Programme) (Latter, PM and DWH Walton. 1988. The cotton strip assay for cellulose decomposition studies in soil, pp. 175-222 to test the decomposition rate in different soil type. In AF Developed by Harrison, PM Latter and DWH Walton (eds.), Cotton Strip Assay: An Index of Decompositionin Soils (Symposium No.24), Institute of Terrestrial Ecology. Degradation was measured by the rate of loss of tensile strength per hour of cotton fabric composed of 96% pure cellulose buried in the soil. In this experiment, 100% unbleached cotton fabric was used. Two identical soil trays, each containing 2 kg of soil (wet wt. Equivalent), were used for each soil sample of LCC and NBC. Cotton strips were prepared in 3 × 8 cm size and sterilized in an autoclave at 121 ° C. for 20 minutes. A soil layer (1 kg wet wt.) Was placed in the tray and the cotton strips were placed on the soil surface consistently and covered with the remaining soil. Four cotton strips were taken over a 30 day period (eg, 15 days and 30 days) to produce a total of eight cotton strip replicas for each harvest in the mixed soil. The tray was incubated at 25 ° C. and sterile distilled water was added to the initial weight every 5 days. The strips were washed with distilled water and air-dried to 0.5 cm on each side to eliminate the “corner” effect. Then dried at 50 ° C. for 12 h. Tensile strength loss was measured in a wrap direction on a precalibrated tensometer (Universal Testing Machine, AGS-5kNJ, Shimadzu, Japan). The results are shown in Table 4 and FIG. 3.

As can be seen in Table 4, it was found that the decomposition rate of cellulose in the LCC soil was increased. In NBC soils, cotton strips hardly decomposed, with only 0.5% and 3% cotton tensile strength loss (CTSL) during the 15 and 30 day incubation periods. However, in the LCC soils, CTSL was 12% and 20% at 15 and 30 day incubation periods, respectively. These results indicate that the enzymatic activity of the microorganisms is associated with the degradation of the organic carbonaceous materials listed in Table 3. In addition, it can be seen that the microbial enzyme activity of the coal abandoned mine soil was induced and stimulated by lichen culture along with the results of Example 4.

Example 6: Literature Production

500 μl of tertiary distilled water was added to a 2 ml microtube to fully release lichen symbiotic algae or cyanobacteria. A plate containing MY medium was inoculated in 16 spots of the released lichen symbiotic algae or cyanobacteria in a constant size of 4 μl each. Inoculated lichen symbiotic algae or cyanobacteria were incubated at 18 ° C. for 2 weeks. After incubation, lichen-forming molds or lumps thereof pulverized on lichen symbiotic algae or cyanobacteria were dentified. Specifically, put about 15 lichen-forming mold in the mortar and then about 300 μl of distilled water was added and ground. About 300 μl of water was added to the finely divided lichen forming mold and mixed well. Next, using a yellow tip cut off the tip was scooped about 8 μl and toothed to lichen symbiotic algae or cyanobacteria. Meanwhile, in the case of lichen-forming molds, the lichen-forming molds of a certain size were placed on lichen-symbiotic algae or cyanobacteria and mixed well.

Thereafter, the toothed lichen forming mold and lichen symbiotic algae or cyanobacteria were incubated for about one month until they were synthesized as one (main body forming) while maintaining a temperature of 15 ° C. Next, all of them were transferred to natural materials such as cork and cultured for 1 month or longer while maintaining the temperature of 15 ° C. During the incubation process, 10 ml of 10% BBM (Bold Basal Medium) was diluted with water to provide a culture solution.

The process of forming a lichen body by synthesizing lichen symbiotic algae or cyanobacteria and lichen liquor forming fungus into one by the above method was observed under a microscope, and the results are shown in FIG. 4. As can be seen in Figure 4, it was confirmed that the lichen body is formed after about one month when the lichen-forming mold is incubated in lichen symbiotic algae or cyanobacteria by the method of the present invention. In addition, it was confirmed that the lichen body was actively breeding over 1 month and 2 months after being transferred to natural objects such as cork. It will therefore be possible to mass produce lichen lichens by this method.

Example 7: Ecological Restoration of Wasteland

The following experiment was conducted to restore the ecology of the wasteland. First, cyanobacteria were incubated for 2 weeks at 20 ° C. in BG11 liquid medium and then centrifuged at 2,000 rpm to obtain only precipitates. The obtained precipitate was mixed with sterilized sodium alginate (alginate sodium) at 121 ° C. for 15 minutes, prepared into a sphere of suitable size (1-20 mm), and left to stand in a 0.1 M CaCl ₂ aqueous solution for 30 minutes to form a particle. After gelation and washing twice with distilled water, alginic acid particles containing cyanobacteria were prepared. The prepared alginic acid particles were placed in a sealed plastic box and stored until use at 4 ° C. The size of the prepared alginic acid particles, the density of the cyanobacteria with the size and time of the alginic acid particles are shown in FIG. As can be seen in Figure 5, as the size of the alginic acid particles it was found that the density of cyanobacteria increases rapidly with time.

Next, the possibility of forming biological soil clusters according to the use of lichens, lichen propagules, algae or cyanobacteria immobilized carriers was examined. Specifically, the dried cyanobacteria alone and PVA mixture, the non-dried cyanobacteria alone and PVA mixture, the alginic acid particles alone and PVA mixture prepared above were evenly dispensed 2 g each on sand and grown at 35 ° C. for 6 months. The formation of biological soil clusters was observed. As a control, distilled water and PVA were used, respectively. In addition, the thickness, compressive strength and water retention of the biological soil cluster formed after a certain period of time was measured. Specific results thereof are shown in FIGS. 6 to 9.

First, as shown in FIGS. 6 and 7, it was confirmed that biological soil clusters were formed when immobilized carriers of sodium alginate or PVA were used. In addition, the formed biological soil clusters showed a certain level of compressive strength (see FIG. 8), and also excellent in water retention (see FIG. 9). In particular, the biomass, such as cyanobacteria, immobilized by the immobilization carrier showed a biomass superior to that of ordinary cyanobacteria, such as non-immobilized (see FIG. 10). It can be seen that the first stage of the lichen can be clearly induced to settle and grow.

According to the present invention, it is possible to restore ecology to a soil in which plants can grow, such as deserts, abandoned mines, oil pollutants, and the like, and thus prevent global warming and environmental and ecological destruction thereof.

Claims (35)

1. (a) immobilizing at least one of lichen symbiotic algae and cyanobacteria by mixing with an immobilization carrier; (b) introducing the immobilized carrier into a wasteland for ecological restoration; (c) inoculating the lichen forming fungus or lichen body on the immobilized carrier; And (d) forming and propagating lichens and growing them into lichens.
2. (a) immobilizing at least one of lichen symbiotic algae and cyanobacteria, lichen forming mold or lichen, by mixing with an immobilizing carrier; (b) introducing the immobilized carrier into a wasteland for ecological restoration; And (c) forming and propagating lichens from the immobilized carrier and growing them into lichens.
3. The method of claim 1 or 2, wherein the alga is trebosia (Trebouxia) Genus, Prédoborussia (Pseudotrebouxia), Sticky caucus (Stichococcus), And Trenteporia (Trentepohlia) Ecological restoration method of barren land, characterized in that any one of the genus.
4. Article according to any one of the preceding claims, cyanobacteria no stock (Nostoc), A ringeu vias (Lyngbya), A micro-collection-house (Microcoleus), An Analog vena (Anabaena), A croissant Rhodococcus (Chrococcus), A Which belongs to the genus Gloecapsa , genus Phormidium , genus Scytonema , genus Synechocystis , genus Calothrix , and genus Tolypothrix . Ecological restoration method of barrens, characterized in that one.
5. The lichen forming fungus of claim 1 or 2, wherein the lichen-forming fungus is Aspicilia species, Caloplaca species, Cladonia species, Collema species, Dematocarpon Dematocarpon species, Diploschistes species, Endocarpon species, Flugensia species, Graphis species , Glypholecia species, Gyrophora Species, Heterodermia species, Physica species, Lecanora species, Phaeophyscia species, P hysconia species, Psora species, Parmelia ), Rhizoplaca species, Ramalina species, Stereocaulon species, Sphaerophorus species, Teloschistes species, Toninia species , Titanium irregularities car Liao (Umbilicaria) species, mouse California (Usnea) species, Philo Forus (Pilophorus) species, Sat Parr Melia (Xanthoparmelia) species, glass thoria (Xanthoria) 1 or more methods ecological restoration of barren, characterized in that belonging to the species selected from the group.
6. The lichen according to claim 1 or 2, wherein the lichen is Aspicilia , Caloplaca , Cladonia , Collema , Dematocarpon Genus, Diploschistes genus, Endocarpon genus, Flugensia genus, Graphis genus, Glypholecia genus, Gyrophora genus, Heterodermia genus, Physica genus, Lecanora genus, Phaeophyscia genus, P hysconia genus, Psora genus, Parmelia genus , Genus Rhizoplaca , genus Ramalina , genus Stereocaulon , genus Sphaerophorus , genus Teloschistes , genus Toninia , genus irregularities car Liao (Umbilicaria), A mouse California (Usnea) in, Philo Forus (Pilophorus), A janto Parr mellitic (Xanthoparmelia), A glass thoria method ecological restoration of barren, characterized in that at least one selected from the group consisting of a lichen (Xanthoria) in lichens.
7. The method of claim 1 or 2, wherein the immobilization support is a method for ecological restoration of barrens, characterized in that the polymer material of the natural polymer.
8. 8. The method of claim 7, wherein the natural polymer has a moisture retention of 15% or more after one day relative to the initial amount of moisture.
9. 8. The method of claim 7, wherein the natural polymer is at least one of alginic acid or salts thereof, cellulose and polysaccharides.
10. The method of claim 1 or 2, wherein the immobilized carrier is an ecological restoration method of barrens, characterized in that contained in an amount of 1.0 to 3.5% by weight relative to the total weight.
11. 3. The method of claim 1 or 2, wherein the step (a) further comprises mixing with the immobilization carrier, and then extruding and molding to prepare and immobilize the molding.
12. The method of claim 1 or claim 2, wherein the barren is any one of the desert, coal mines, oil pollution, volcanic ash, hazardous mineral disposal site, industrial waste disposal site, rock and bald mountain. Method of ecological restoration of wasteland.
13. The method of claim 1 or 2, wherein the introduction is a method of ecological restoration of the wasteland, characterized in that the immobilization carrier is arranged to a depth of 1 cm or less from the surface or the surface of the wasteland.
14. The method of claim 1, wherein the lichen-forming mold of step (c) is inoculated with 1 to 20 cells per unit area (cm ² ) ecological restoration method of barrens.
15. The method of claim 1, wherein the limb of step (c) is inoculated in 1 to 20 individuals per unit area (cm ² ) ecological restoration method of barrens.
16. The method of claim 1, wherein the steps (b) and (c) is the ecological restoration method of the barren, characterized in that the order is reversed.
17. The method of claim 1 or 16, further comprising the step of applying a coagulant at a thickness of 0.5 to 2 times the diameter of the immobilized carrier in the last step of the step.
18. 18. The method of claim 17, wherein the coagulant is polylactide, polyglycolide, poly (lactide-co-glycolide), poly-β-hydroxy butyl acid, poly anhydride, polyorthoester, polyurethane, poly Amide polycarbonate, polyethylene, polypropylene, poly (ethylene glycol), poly (ethylene oxide), poly (ethylene terraphthalate), polyvinyl alcohol, polyvinyl ether, polyvinyl ester, poly (vinylchloride), polyvinylpyrroli Don, polysiloxane, poly (vinyl alcohol), poly (vinyl acetate), polystyrene, polyurethane and copolymers thereof, derived cellulose such as alkyl cellulose, hydroalkyl cellulose, cellulose ether, cellulose ester, nitrocellulose, methyl cellulose, ethyl Cellulose, hydropropyl cellulose, hydroxy-propyl methyl cellulose, Synthetic cellulose, polymers of acrylic acid, esters thereof, such as hydroxybutyl methyl cellulose, cellulose acetate, cellulose propinate, cellulose acetate butylate, cellulose acetate phthalate, carboxyethyl cellulose, cellulose triacetate and cellulose cellulose sodium salt Derivative copolymers or methacrylates, poly (methyl methacrylate), poly (ethyl methacrylate), poly (butyl methacrylate), poly (isobutyl methacrylate), poly (hexyl methacrylate), poly (Isodecylmethacrylate), poly (isopropylacrylate), poly (isobutylacrylate) and poly (octadecylacrylate), poly (butyl acid), poly (valeric acid) and poly (lactide- Co-caprolactam), copolymers and mixtures thereof Ecological restoration method of barren comprising or higher.
19. A composition for ecological restoration of wasteland comprising at least one of lichen symbiotic algae and cyanobacteria, lichen forming mold or lichen body, and immobilized carrier.
20. 20. The genus of claim 19, wherein the algae are of the genus Trebouxia, Pseudotrebouxia,Stichococcus), And ecological restoration composition of the barren land, characterized in that any one of the genus Trentepohlia.
21. 20. The genus Cyanobacteria according to claim 19, wherein the cyanobacteria is in the genus Nostoc , in the genus Lyngbya , in the genus Microcoleus , in the genus Anabaena , in the genus Crocoker tm, Chrococcus Gloecapsa genus, Phormidium genus, Scytonema genus, Synechocystis genus, Calothrix genus, and Tolypothrix genus A composition for ecological restoration of wasteland.
22. 20. The lichen-forming fungus of claim 19, wherein the lichen-forming fungus is Aspicilia species, Caloplaca species, Cladonia species, Collema species, Dematocarpon species, Diploschistes species, Endocarpon species, Flugensia species, Graphis species , Glypholecia species, Gyrophora species, heteroder Heterodermia species, Physica species, Lecanora species, Phaeophyscia species, P hysconia species, Psora species, Parmelia species, Lyzo Rhizoplaca species, Ramalina species, Stereocaulon species, Sphaerophorus species, Teloschistes species, Toninia species, Umbrica Umbilicaria species, Usnea species, Pilophorus species, Zantoparmel ( Xanthoparmelia ) species, Xanthoria species ( Xanthoria ) A species for ecological restoration of barrens, characterized in that selected from the group belonging to.
23. 20. The lichen according to claim 19, wherein the lichen is Aspicilia , Caloplaca , Cladonia , Collema , Dematocarpon , Dipro . The genus Diploschistes , genus E ndocarpon , genus Flugensia , genus Graphis, genus Glypholecia , genus Gyrophora , heterodermia ( Heterodermia genus, Physica genus, Lecanora genus, Phaeophyscia genus, P hysconia genus, Psora genus, Parmelia genus, Lyzopraca ( Rhizoplaca ), Ramalina genus, Stereocaulon genus, Sphaerophorus genus, Teloschistes genus, Toninia genus, Umbiricaria Umbilicaria genus, Usnea genus, Pilophorus genus, Xanthoparm elia ) genus, Xanthoria ( Xanthoria ) belonging to any one or more lichens selected from the group consisting of lichens, the composition for ecological restoration of barrens.
24. 20. The composition of claim 19, wherein the immobilized carrier is a polymer material of a natural polymer.
25. The composition of claim 24, wherein the natural polymer has a water retention of 15% or more after one day relative to the initial amount of moisture.
26. 25. The composition of claim 24, wherein the natural polymer is at least one of alginic acid or salts thereof, cellulose and polysaccharides.
27. 20. The composition of claim 19, wherein the immobilized carrier is contained in an amount of 1.0 to 3.5% by weight relative to the total weight.
28. 20. The composition of claim 19, wherein the composition further comprises a coagulant.
29. 29. The method of claim 28, wherein the coagulant is polylactide, polyglycolide, poly (lactide-co-glycolide), poly-β-hydroxy butyl acid, poly anhydride, polyorthoester, polyurethane, poly Amide polycarbonate, polyethylene, polypropylene, poly (ethylene glycol), poly (ethylene oxide), poly (ethylene terraphthalate), polyvinyl alcohol, polyvinyl ether, polyvinyl ester, poly (vinylchloride), polyvinylpyrroli Don, polysiloxane, poly (vinyl alcohol), poly (vinyl acetate), polystyrene, polyurethane and copolymers thereof, derived cellulose such as alkyl cellulose, hydroalkyl cellulose, cellulose ether, cellulose ester, nitrocellulose, methyl cellulose, ethyl Cellulose, hydropropyl cellulose, hydroxy-propyl methyl cellulose, Synthetic cellulose, polymers of acrylic acid, esters thereof, such as hydroxybutyl methyl cellulose, cellulose acetate, cellulose propinate, cellulose acetate butylate, cellulose acetate phthalate, carboxyethyl cellulose, cellulose triacetate and cellulose cellulose sodium salt Derivative copolymers or methacrylates, poly (methyl methacrylate), poly (ethyl methacrylate), poly (butyl methacrylate), poly (isobutyl methacrylate), poly (hexyl methacrylate), poly (Isodecylmethacrylate), poly (isopropylacrylate), poly (isobutylacrylate) and poly (octadecylacrylate), poly (butyl acid), poly (valeric acid) and poly (lactide- Co-caprolactam), copolymers and mixtures thereof Ecological restoration composition of the barren, characterized in that or more.
30. (a) artificially culturing lichen symbiotic algae or cyanobacteria; (b) inoculating the lichen forming fungus on the alga or cyanobacteria to form lichens; And (c) inducing the propagation of lichen lichens by placing the algae or cyanobacteria on which the lichen bodies are formed in a substance according to the growth environment of the algae or cyanobacteria.
31. The genus of claim 30, wherein the algae are of the genus Trebouxia, Pseudotrebouxia,Stichococcus), And a mass belonging to the genus Trentepohlia.
32. 31. The method of claim 30, cyanobacteria no stock (Nostoc), A ringeu vias (Lyngbya), A micro-collection-house (Microcoleus), An Analog vena (Anabaena), A croissant coker tm (Chrococcus), A cap for him One of the genus Gloecapsa , Phormidium , Scytonema , Synechocystis , Calothrix , and Tolypothrix . A mass production method of lichen lichen body characterized by the above-mentioned.
33. 31. The lichen forming fungus of claim 30, wherein the lichen-forming fungus is Aspicilia species, Caloplaca species, Cladonia species, Collema species, Dematocarpon species, Diploschistes species, Endocarpon species, Flugensia species, Graphis species , Glypholecia species, Gyrophora species, heteroder Heterodermia species, Physica species, Lecanora species, Phaeophyscia species, P hysconia species, Psora species, Parmelia species, Lyzo Rhizoplaca species, Ramalina species, Stereocaulon species, Sphaerophorus species, Teloschistes species, Toninia species, Umbrica Umbilicaria species, Usnea species, Pilophorus species, Zantoparmel Liao (Xanthoparmelia) species, glass thoria (Xanthoria) method mass production of lichens magazine body, characterized in that at least one member selected from the group belonging to the species.
34. 31. The method for mass production of lichen lichens according to claim 30, wherein the lichen forming mold of step (b) is inoculated with 1 to 20 cells per unit area (cm ² ).
35. 31. The method for mass production of lichen limbs according to claim 30, wherein the article is any one of wood, stone, rock, cork or sand.

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