WO2012008634A1 - Procédé de production de masse du lichen tali, procédé de restauration de systèmes écologiques au moyen du tali produit, et composition pour la restauration de systèmes écologiques - Google Patents

Procédé de production de masse du lichen tali, procédé de restauration de systèmes écologiques au moyen du tali produit, et composition pour la restauration de systèmes écologiques Download PDF

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WO2012008634A1
WO2012008634A1 PCT/KR2010/004627 KR2010004627W WO2012008634A1 WO 2012008634 A1 WO2012008634 A1 WO 2012008634A1 KR 2010004627 W KR2010004627 W KR 2010004627W WO 2012008634 A1 WO2012008634 A1 WO 2012008634A1
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species
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lichen
poly
cellulose
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PCT/KR2010/004627
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Korean (ko)
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허재선
류연풍
고영진
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순천대학교 산학협력단
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Priority to PCT/KR2010/004627 priority Critical patent/WO2012008634A1/fr
Priority to KR1020137000891A priority patent/KR101543134B1/ko
Priority to CN201080068074.9A priority patent/CN103153495B/zh
Publication of WO2012008634A1 publication Critical patent/WO2012008634A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/12—Unicellular algae; Culture media therefor
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
    • B09C—RECLAMATION OF CONTAMINATED SOIL
    • B09C1/00—Reclamation of contaminated soil
    • B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
    • B09C—RECLAMATION OF CONTAMINATED SOIL
    • B09C1/00—Reclamation of contaminated soil
    • B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
    • B09C1/105—Reclamation of contaminated soil microbiologically, biologically or by using enzymes using fungi or plants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
    • Y02W10/00—Technologies for wastewater treatment
    • Y02W10/10—Biological treatment of water, waste water, or sewage

Definitions

  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • plants such as soil crops stop growing and cause secondary pollution such as groundwater pollution.
  • 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.
  • 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.
  • the present invention provides a method for restoring the ecology of barren land using lichen symbiotic algae, cyanobacteria, lichen forming fungi, or lichen bodies.
  • lichen lichen refers to a symbiotic complex of lichen-forming fungi and algae and / or cyanobacteria.
  • 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.
  • 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.
  • the first step is to immobilize (a) at least one of lichen symbiotic algae and cyanobacteria by mixing with an immobilization carrier.
  • lipid symbiotic 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 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 (Gloecap
  • 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.
  • 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.
  • the alginic acid salt preferably has a molecular weight of 10,000 to 1,000,000 Da.
  • 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).
  • 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.
  • (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).
  • 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.
  • plants can grow in neutral or weakly acidic to weak alkalinity.
  • 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.
  • 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.
  • the immobilized carrier 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • step (b) is introduced into the barren for the purpose of ecological restoration.
  • 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.
  • 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- ⁇ -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
  • 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).
  • 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.
  • 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.
  • 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.
  • the process of changing the barren soil into soil where plants can grow is called cloth.
  • lichens invade, form clusters, and organic matters accumulate in the soil.
  • 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.
  • 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.
  • the present invention provides a composition for the ecological restoration of barrens to be used in the method of ecological restoration of the barrens.
  • 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.
  • 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.
  • lichens have very small propagules and can settle anywhere except on extremely flat surfaces. Many lichens have other advantages as early invaders.
  • 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.
  • 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.
  • 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).
  • 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).
  • 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.
  • 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.
  • the limb can be produced by the mass production method of the limb as described below.
  • 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.
  • the soil pH was higher in LCC soils than in NBC soils, as compared to lichen-colonized coalmine (LCC) and noncolonized bare coalmine (NCC).
  • LCC lichen-colonized coalmine
  • NCC noncolonized bare coalmine
  • 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.
  • LCC lichen-colonized coalmine
  • NCC noncolonized bare coalmine
  • 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.
  • the bioremediation soil modifying agent of the present invention was confirmed to form biological soil clusters.
  • 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.
  • the present invention provides a method for mass production of lichens of lichens.
  • 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.
  • lichen lichen refers to a symbiotic complex of lichen-forming fungi and algae and / or cyanobacteria.
  • 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.
  • the first step is artificially culturing lichen symbiotic algae or cyanobacteria.
  • 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.
  • Transistor tepo Liao in such Tina (Trentepohlia abietina), ah rugi labor (Trentepohlia aeruginosa), Aurea (Trentepohlia aurea), are Bohrium (Trentepohlia arborum) in the AVI .
  • 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 mur) FORT Medium (Phormidium) in, ski weekends and Cinema Now Pony Qom (Scytonema japonicum) in such a thereof Scytonema genus, Synechocy
  • 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
  • the next step is to inoculate lichen-forming mold to algae or cyanobacteria cultured in the first step to form lichen body.
  • 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.
  • “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.
  • the tooth lichen-forming mold to be toothed has a diameter of 2 mm or less, more preferably 1 to 2 mm per cell.
  • a part of algae or cyanobacteria and a part of lichen forming molds are ecologically combined, and the combination of lichens is formed.
  • a large amount of the limb is formed by the method of germination, heat germination or nutrition propagation.
  • 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.
  • 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.
  • 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.
  • lichens are multiplyed and lichen bodies are formed in a large amount so that lichen bodies can be used in large quantities. do.
  • 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).
  • 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.
  • 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.
  • 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.
  • 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.
  • FIG. 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.
  • FIG. 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.
  • lichens from China's Shapotou and Inner Mongolia deserts were collected.
  • the collected lichens were analyzed as Aspicilia , Caloplaca , Cladonia , Collema , Dematocarpon , Depro
  • the results are mean and standard deviation for eight replicates for each lichen.
  • LCC soils Lichen-colonized coalmine (LCC) soils and noncolonized bare coalmine (NCC) soils in the coal waste mine of Example 1 were 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.
  • 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).
  • the results are mean and standard deviation of 10 samples for each 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.
  • one-way-ANOVA SPSS Version 11.0
  • 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 ⁇ -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).
  • 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.
  • 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.
  • Table 4 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.
  • 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.
  • 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.
  • 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.
  • ⁇ l 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 ⁇ l 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 ⁇ l of distilled water was added and ground.
  • lichen-forming molds About 300 ⁇ l 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 ⁇ l 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.
  • 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.
  • BBM Back Basal Medium
  • FIG. 4 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.
  • 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.
  • 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.
  • FIGS. 6 and 7 it was confirmed that biological soil clusters were formed when immobilized carriers of sodium alginate or PVA were used.
  • the formed biological soil clusters showed a certain level of compressive strength (see FIG. 8), and also excellent in water retention (see FIG. 9).
  • 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.

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Abstract

La présente invention concerne un procédé de restauration du système écologique de terrains inutilisables tels que les déserts, les mines de charbon abandonnées, les régions polluées pas les hydrocarbures, les régions affectées par les volcans, les régions où des minéraux dangereux ou des déchets industriels ont été jetés, les terres rocailleuses et les montagnes nues, au moyen d'algues avec lesquelles les lichens entretiennent une relation symbiotique, de cyanobactéries, de champignons formant les lichens, ou de tali; ainsi qu'une composition de restauration du système écologique utilisée dans ce procédé. Par ailleurs, l'invention concerne un procédé de production de masse de tali nécessaire à la restauration du système écologique de terrains inutilisables. De plus, l'invention peut prévenir la désertification et les tempêtes de poussière jaune, et permettre aux régions polluées telles que les mines de charbon abandonnées et les régions polluées par les hydrocarbures de se reconstituer et d'être plus favorables à la croissance de plantes, de façon à empêcher la pollution environnementale et écologique dans les déserts, les mines de charbon abandonnées et les régions polluées par les hydrocarbures.
PCT/KR2010/004627 2010-07-15 2010-07-15 Procédé de production de masse du lichen tali, procédé de restauration de systèmes écologiques au moyen du tali produit, et composition pour la restauration de systèmes écologiques WO2012008634A1 (fr)

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CN110663439A (zh) * 2019-11-08 2020-01-10 攀枝花学院 用鸡蛋花对金沙江干热河谷地区进行生态修复的方法
CN114574474A (zh) * 2020-12-02 2022-06-03 中国科学院微生物研究所 一种便携式生物结皮种源及其制备方法
CN115247132A (zh) * 2021-04-26 2022-10-28 中国科学院微生物研究所 一种组合型人工生物结皮治理荒漠的方法
CN115843485A (zh) * 2022-12-09 2023-03-28 国能宝日希勒能源有限公司 铁尾矿基质的生物改良方法及应用

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KR101683842B1 (ko) * 2014-07-30 2016-12-07 순천대학교 산학협력단 조류를 이용한 생태복원용 블록의 제조방법
KR101705578B1 (ko) * 2014-10-20 2017-02-22 순천대학교 산학협력단 생태복원용 조성물 및 이를 이용한 생태복원 방법
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CN102757792B (zh) * 2012-06-28 2015-09-09 环境保护部华南环境科学研究所 一种生活污泥制成的复合生态固沙剂及其制备方法与应用
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CN114574474A (zh) * 2020-12-02 2022-06-03 中国科学院微生物研究所 一种便携式生物结皮种源及其制备方法
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CN115843485A (zh) * 2022-12-09 2023-03-28 国能宝日希勒能源有限公司 铁尾矿基质的生物改良方法及应用

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