GB2597602A

GB2597602A - Method for applying moss sporophore suspension containing chitosan/glucan in greenery coverage of bare land

Info

Publication number: GB2597602A
Application number: GB2109405.7A
Authority: GB
Inventors: Sun Geng; Lei Yanbao; Chen Ke; Xia Hongxia; Zhu Dalin; Ren Hui
Original assignee: Chengdu Institute of Biology of CAS; Southwest University of Science and Technology
Current assignee: Chengdu Institute of Biology of CAS; Southwest University of Science and Technology
Priority date: 2020-11-06
Filing date: 2021-06-30
Publication date: 2022-02-02
Anticipated expiration: 2041-06-30
Also published as: CN112335508A; GB202109405D0; GB2597602B; CN112335508B

Abstract

The present invention discloses a method for applying a moss sporophore suspension containing chitosan and glucan in greenery coverage of a bare land. The method includes the steps of: collecting moss spores and adding a hydrogen peroxide solution to the spores to obtain a mixed solution of disinfected spores and hydrogen peroxide; inoculating the mixed solution of disinfected spores and hydrogen peroxide onto a spore germination-inducing culture medium for spore germination-inducing culture; adding the cultured spores to a solid subculture medium for subculturing; mixing the subcultured spores with a fluid medium for shaking culture to obtain a dark green suspension; adding chitosan, glucan, ammonium ferric citrate, phenoxyacetic acid, zinc sulfate and potassium nitrate to the dark green suspension to obtain a moss sporophore suspension; evenly painting a moss adhesive on a bare land, and then evenly spraying the moss sporophore suspension on a surface of the moss adhesive, covering the surface with a sunshading net and performing spray irrigation in time. The moss sporophore suspension serves for the greenery coverage of a bare land and has a high greenery coverage rate after growth.

Description

METHOD FOR APPLYING MOSS SPOROPHORE SUSPENSION CONTAINING

CHITOSAN/GLUCAN IN GREENERY COVERAGE OF BARE LAND

Technical Field

The present invention relates to the technical field of greenery coverage of a bare land, and in particular to a method for applying a moss sporophore suspension containing chitosan/glucan in greenery coverage of a bare land.

Background

Currently, bryophyte is a kind of ordinary higher plant and widely distributed in nature. Bryophyte has clumping and dormant habits, strong water absorbing capacity, rapid growth rate and strong cold resistance; and is less liable to get sick to cause insect attack. It is a kind of pioneer plant capable of rapidly achieving fission in an appropriate environment. Bryophyte can absorb dust particles accumulated in the air and other substances to fa soil, and can further change the ambient humidity, pH value and other indicators, thus forming soil suitable for the survival of other higher plants. Thus, bryophyte can improve the environment where vegetation cannot grow.

With the development of economy, land afforestation and beautification becomes a higher pursuit of life. Bare land existing in large numbers, including bare rock, bare ground, bare sandy land, bare project profile, bare land polluted by heavy metals, bare land polluted by pesticides, radioactive source-polluted soil, and the like, is the greening target area highly concerned by people. Since bare land has no soil coverage and the environment is extremely severe, there is no change several years and even several decades later if the bare land is not interfered manually. At present, there are a large number of bare land due to the reasons, such as, natural environment, ore extraction, and urbanization construction, which not only influences the city landscape, but also easily leads to water and soil loss, restriction of the regional development and other problems. Measures, such as, net hanging, soil-spraying technology, cultivation of hanging plants and climbing plants in the prior art are used for rock greening, but the majority of plants cannot grow for a long time, or have undesirable greening effect. Bryophyte can green the rock to solve the problem of greening rock slope and the like. Bryophyte is short and small, and has a simple morphological structure, no real root system and thus, has a very limited ability to regulate and control water dredging. Due to its specific structure and physiological adaptive mechanism, bryophyte can grow and reproduce in high temperature, drought, extremely cold, frozen, low light and other extreme environment where other terrestrial plants are hard to survive. Many kinds of these microphanerophyte are pioneer species in the primary succession stage and usually, these pioneer species are clustered in open/barren areas and other places where tracheophyte could not survive, especially in the area where a large number of rocks are bare; other plants cannot survive, but bryophyte has unique advantages in these areas. With the development of moss greening project, the cultivation and processing technology of moss materials are more important, especially in the aspect of cultivating moss sporophore. Due to lack of nutrition, the moss sporophore will lead to the failure of greenery coverage when bonded on a bare land, meanwhile, there are limitations in the solid adhesive used, and partial adhesives give off irritating odors or contain poisonous and harmful ingredients; moreover; these adhesives cannot resist outdoor wind blowing and thus have poor bonding effect on moss.

Summary

An objective of the present invention is to solve at least the above problems and/or defects, and to provide the following advantages to be specified hereafter.

To achieve these objectives and advantages of the present invention, provided is a method for applying a moss sporophore suspension containing chitosan/glucan in greenery coverage of a bare land, including the following steps: step I, collecting capsula of moss, and cutting the capsula to collect spores when calyptra of the capsula begins falling off, using distilled water to elute spores residual on the capsula; adding a 3-4%why hydrogen peroxide solution to the spores, and shaking for 6-10 min to obtain a mixed solution of disinfected spores and hydrogen peroxide; where a volume ratio of the spores to the 3-4%w/w hydrogen peroxide solution is 1:10; step II, inoculating the mixed solution of disinfected spores and hydrogen peroxide onto a spore germination-inducing culture medium for spore germination-inducing culture for 30-50 d to obtain a suspension containing endospores and protonema; step Ill, taking and adding the suspension containing endospores and protonema onto a solid subculture medium for subculturing for 28-32 d; thus obtaining a suspension containing green endospore colonies and protonema.

step IV, taking the suspension containing green endospore colonies and protonema in the step HT and mixing with a fluid medium to obtain a mixed solution, then subjecting the mixed solution to shaking culture for 25-30 d, thus obtaining a dark green suspension; step V adding chitosan, glucan, ammonium ferric citrate. phenoxyacetic acid, zinc sulfate and potassium nitrate to the dark green suspension in the step IV, and stirring them to obtain a moss sporophore suspension; step VI, evenly painting a moss adhesive onto a bare land, and evenly spraying the moss sporophore suspension on a surface of the moss adhesive; covering the surface with a sunshading net and performing spray irrigation in time, such that a surface layer of the polluted bare land keeps a humidity within 20-25%.

Preferably, in the step II, conditions of the spore germination-inducing culture are as follows: tLmperature is 22-28°C, illumination intensity is 2500-30001x, illumination time during the first 1-5 d is 20-24 h/d, illumination time after the 6th day is 8 hid: the moss spore germination-inducing culture medium includes as follows: MS + 0.5-1.0 mg/LKT + 0.2 mg/L GA3 + 20 g/L white sugar with a pH value of 6.0; and a volume ratio of the mixed solution of disinfected spores and hydrogen peroxide to the spore germination-inducing culture medium is 1:5.

Preferably, in the step ITT, the subculturing conditions are as follows: a temperature is 22-28°C, an illumination intensity is 2500-3000h, an illumination rime is 8 hid: and the solid subculture medium includes as follows: MS + 0.5-1.0 mg/L BA + 0.2 mg,/ 2, 4-D + 0.3 mg/L TAA + 20 g/L white sugar with a pH value of 6.0.

Preferably, in the step TV, the shaking culture is performed on a shaker at 100 r/min; the shaking culture conditions on the shaker are as follows: temperature is 22-28°C, illumination intensity is 2500-30001x, illumination time is 8 hid: a dark green suspension is obtained until the mixed solution thickens and turns dark green; and the fluid medium: MS + 0.1 mg/L BA + 0.2-0.5 mg/L NAA + 20 g/L white sugar with a pH value of 6.0.

Preferably, in the step V, according to parts by weight, 0.5-1 parts of chitosan, 0.3-0.5 parts of glucan, 02-0.4 parts of ammonium ferric citrate, 0.1-0.3 parts of phenoxyacetic acid, 0.1-02 parts of zinc sulfate and 0.1-0.3 parts of potassium nitrate are added to 100 parts of the dark green suspension in the step IV; and then stirring is performed to obtain a moss sporophore suspension.

Preferably, the bare land is any one of a bare rock, a bare ground, a bare sandy land, a bare project profile, a bare land polluted by heavy metals, a bare land polluted by pesticides, and a radioactive source-polluted soil.

Preferably, a preparation method of the toss adhesive is as follows: according to parts by weight, 10-30 parts of melamine, 10-20 parts of urea, 60-90 parts of attapulgite and 50-80 parts of sepiolite powder are added to a ball mill pot; and a milling ball is added to the ball mill pot and liquid nitrogen is fed to the ball mill pot, such that the materials are immersed into the liquid nitrogen at a temperature within -160°C to -175°C, and the volatile amount and feed amount of the liquid nitrogen are kept balance, so as to stabilize the liquid level; ball milling is performed for 30-45min after keeping constant temperature for 15 min; after the ball milling is completed, the ball mill pot is moved to a vacuum glove box, and placed for 3-5 h to collect ball milling materials; step 11, according to parts by weight, 50-100 parts of konjac glucomannan, 50-100 parts of chitosan and 50-100 parts of modified collagen are taken and added to 200-300 parts of water, and stirred for 30-60 mm at a rate of 1000-1200 r/min; half of the ball milling materials is added during the stirring and mixing process at a rate of 10-30 parts/min; then 40-60 parts of latex powder, 10-30 parts of stearic acid and 5-15 parts of ammonium zirconium carbonate are added, mixed and stirred continuously for 30-45 min at a revolving rate of 1000-1200 r/min; meanwhile, the remaining half of the ball milling materials is added at a rate of 10-30 parts/min during the stirring arid mixing process to obtain a dedicated moss adhesive; Preferably, the latex powder is one or more than two of ethylene-vinyl acetate copolymer latex powder, acrylate-styrene copolymerized rubber powder, ternary-copolymerized rubber powder of vinyl acetate, acrylate and higher fatty acid vinyl ester.

Preferably, the collagen is replaced with a modified collagen; and a preparation method thereof is as follows: according to parts by weight, 50-100 parts of collagen are added to 1000-1200 water for ultrasonic treatment for 30-45 min; 5-8 parts of catcchinic acid was added and then ultrasonic treatment was performed for 15-30 min; then 15-25 parts of poly L-glutamic acid are added: and then the mixed materials are heated up to 45-60°C:, and irradiated by UV light, stirred for reaction for 15-30 min, and subjected to centrifugal separation, thus obtaining the modified collagen.

Preferably, the ultrasonic treatment has a power of 80-120 W and a frequency of 30-50 kHz; and the UV light has an irradiation power of 50-500W, and the UV light has a wavelength of 220400 nm.

The present invention at least includes the following beneficial effects: the present invention obtains a nutritious moss sporophore suspension by collecting capsula of moss and spores, successively performing spore germination-inducing culture, subculturing and fluid medium culture, and finally mixing the obtained suspension with nutrients. The moss sporophorc suspension is sprayed on a surface of a moss adhesive for greenery coverage of a bare land; the greenery coverage rate is high after growth, and the moss adhesive used has good bonding effect to the moss, is eco-friendly and causes no harm to bryophyte. Meanwhile, on the one hand, die konjac glucomannan, chitosan, collagen, attapulgite and sepiolite powder contained therein can provide nutrients for moss, on the other hand, it has thickening, thixotropy and moisture-holding effects, which can improve the bonding force mid air permeability of the adhesive to promote the rapid plantation of the moss on bare rock, rubble, overturned tree or bare ground and other severe environment.

Other partial advantages, objectives and features of the present invention will be embodied by the following description; a portion of them will be further understood by a person skilled in the art by the research and practice on the present invention.

Detailed Description of the Embodiments

The present invention will be further described specifically in combination with the examples hereafter, such that a person skilled in the art can implement the examples based thereon by reference to the text of the description.

It should be understood that the terms such as, "including", "containing" and "comprising" used herein do not exclude the existence or addition of one or more of other elements or combinations thereof.

The dedicated moss adhesive prepared in Examples 1-6 is subjected to performance test; where the measurement standard of tensile shear strength is GB/T7124-2008, and the measurement standard of peel strength is GRIT 779 I -1995.

Example 1:

A method for applying a moss sporophore suspension containing chitosan/glucan in greenery coverage of a bare land, included the following steps: step 1, capsula of Racomitrium japonicum was collected, and the capsula was cut to collect spores when calypira of the capsula began falling off, distilled water was used to elute spores residual on the capsula; a Iriow/w hydrogen peroxide solution was added to the spores, and shaken for 6 min to obtain a mixed solution of disinfected spores and hydrogen peroxide; where a volume ratio of the spores to the 31Nwyky hydrogen peroxide solution was 1:10; step 11, 2 mL mixed solution of disinfected spores and hydrogen peroxide was inoculated onto a 10 iriL spore gennination-inducing culture medium for spore germination-inducing culture for d to obtain a suspension containing spores and protonema; conditions of the spore germination-inducing culture were as follows: temperature was 22°C, illumination intensity was 25001x, illumination time during the first 1-5 d was 20 h/d, illumination time after the 6th day was 8 h/d; the moss spore germination-inducing culture medium included as follows: an MS medium + 0.5 mg/L KT (kinctin) + 0.2 mg/L GA3 (gibbercllin) + 20 g/L white sugar with a pH value of 6.0; step 111, 2 mL of the suspension containing spores and protonema in the step 11 was taken and added to a 10 mL solid subculture medium for subculturing for 28 d to obtain a suspension containing green spore colonies and protonema; the subculturing conditions were as follows: a temperature was 22°C, an illumination intensity was 25001x, an illumination time was 8 h/d; and the solid subculture medium included as follows: an MS medium + 0.5 mg/L BA (benzylamino adenine) + 0.2 mg/L 2,4-D(24-dichlorphenoxyacetic acid) + 0,3 mg/L TAA (auxin) + 20 g/L white sugar with a pH value of 6.0; step IV, 2 mL of the suspension containing green spore colonies and protonema in the step III was taken and mixed with a 10 mL fluid medium to obtain a mixed solution; then the mixed solution was subjected to shaking culture for 25 d on a shaker at a rotating rate of 100 r/min to obtain a dark green suspension; the shaking culture conditions on the shaker are as follows: tunperature was 22°C, illumination intensity was 25001x, illumination time was 8 hid; the dark green suspension was obtained until the mixed solution thickened and tamed dark green; and the fluid medium: an MS medium + 0.1 mg/L BA + 0.2 mg/L NAA (naphthylacetic acid) + 20 g/L white sugar with a pH value of 6.0; step V, 0.5 g chitosan, 0.3 g glucan, 0.2 g ammonium ferric citrate, 0.1 g phenoxyacetic acid, 0.1 g zinc sulfate and 0.1 g potassium nitrate were added to 10 g dark green suspension in the step IV, and stirred to obtain a moss sporophore suspension; step VI, a moss adhesive was evenly painted onto a bare rock, and the moss sporophore suspension was evenly sprayed on a surface of the moss adhesive; the surface was covered with a sunshading net and subjected to spray irrigation in time, such that a surface layer of the polluted bare land kept a humidity of 20%; after 30 d incubation, the coverage of moss was up to 95%; and after 60 d incubation, it was not found that the moss layer was separated from the rock; a preparation method of the toss adhesive was as follows: 18 g melamine, 15 g urea, 75 g attapulgitc and 72 g sepiolite powder were added to a bail mill pot; and a milling ball was added to the ball mill pot and liquid nitrogen was fed to the ball mill pot, such that the materials were immersed into the liquid nitrogen; the temperature was -170°C., and the volatile amount and feed amount of the liquid nitrogen were kept balance, so as to stabilize the liquid level; ball milling was performed for 40 min after keeping constant temperature for 15 min; after the ball milling was completed, the ball mill pot was moved to a vacuum glove box, and placed for 3 h to collect ball milling materials; step 11, 80 g konjac glucomannan, 78 g chitosan and 75 g collagen were taken and added to 245 g water, and stirred for 60 min at a rate of 1200 r/min: half of the ball milling materials was added during the stirring and mixing process at a rate of 20 g/min; then 55 g ethylene-vinyl acetate copolymer latex powder, 25 g stearic acid and 12 g ammonium zirconium carbonate were added, mixed and stirred continuously for 30 min at a rotating rate of 1200 r/min; meanwhile, the remaining half of the ball milling materials was added at a rate of 20 g/min during the stirring and mixing process to obtain a moss adhesive; the adhesive had a tensile shear strength of 10.6 Mpa, and a peel strength of 11.8N/mm.

Example 2:

A method for applying a moss sporophore suspension containing chitosan/glucan in greenery coverage of a bare land, included the following steps: step I, capsula of Racomitrium japonicum was collected, and the capsula was cut to collect spores when calyptra of the capsula began falling off, distilled water was used to elute spores residual on the capsula; a 4%w/w hydrogen peroxide solution was added to the spores, and shaken for 10 min to obtain a mixed solution of disinfected spores and hydrogen peroxide; where a volume ratio of the spores to the 4%,w/w hydrogen peroxide solution was 1:10; step II, 2 mL mixed solution of disinfected spores and hydrogen peroxide was inoculated onto a 10 mL spore germination-inducing culture medium for spore germination-inducing culture for 50 d to obtain a suspension containing spores and protonema, conditions of the spore gemination-inducing culture were as follows: temperature was 28°C, illumination intensity was 30001x, illumination time during the first 1-5 d was 24 h/d, illumination time after the 6th day was 8 hid; the moss spore germination-inducing culture medium included as follows: an MS medium + 1.0mg/L KT (kinetin) + 0.2 mg/L GA3 (gibberellin) + 20 g/L white sugar with a pH value of 6.0; step HT, 2 mL of the suspension containing spores and protonema in the step IT was taken and added to a 10 mL solid subculture medium for subculturing for 32 d to obtain a suspension containing green spore colonies and protoncma, the subculturing conditions were as follows: a temperature was 28°C, an illumination intensity was 3000k an illumination time was 8 h/d; and the solid subculture medium included as follows: an MS medium + 1.0mg/L BA (benzylamino adenine) + 0.2 mg/L 2,4-D(2,4-dichlorphenoxyacetic acid) + 0.3 mg/L IAA (auxin) + 20 g/L white sugar with a pH value of 6.0; step IV, 2 mL of the suspension containing green spore colonies and protonema in the step 111 was taken and mixed with a 10 mL fluid medium to obtain a mixed solution; then the mixed solution was subjected to shaking culture for 25 d on a shaker at a rotating rate of 100 r/min Lo obtain a dark green suspension; the shaking culture conditions on the shaker are as follows: temperature was 28°C, illumination intensity was 30001x, illumination time was 8 hid; the dark green suspension was obtained until the mixed solution thickened and turned dark green; and the fluid medium: an MS medium + 0.1 mg/L BA + 0.5mg/L NAA (naphthylacetic acid) + 20 g/L white sugar with a pH value of 6.0; step V. 1 g chitosan, 0.5 g glucan, 0.4 g ammonium ferric citrate, 0.3 g phenoxyacetic acid, 0.2 g zinc sulfate and 0.3 g potassium nitrate were added to 10 g dark green suspension in the step IV, and stirred to obtain a moss sporophore suspension; step VI, a moss adhesive was evenly painted onto a bare rock, and the moss sporophore suspension was evenly sprayed on a surface of the moss adhesive; the surface was covered with a sunshading net and subjected to spray irrigation in time, such that a surface layer of the polluted bare land kept a humidity of 25%; after 30 d incubation, the coverage of moss was up to 94%; and after 60 d incubation, it was not found that the moss layer was separated from the rock; a preparation method of the toss adhesive was as follows: 18 g melamine, 15 g urea, 75 g attapulgite and 72 g sepiolite powder were added to a ball mill pot; and a milling ball was added to the ball mill pot and liquid nitrogen was fed to the ball mill pot, such that the materials were immersed into the liquid nitrogen; the temperature was -170°C, and the volatile amount and feed amount of the liquid nitrogen were kept balance, so as to stabilize the liquid level; ball milling was performed for 40 mm after keeping constant temperature for 15 min; after the ball milling was completed, the ball mill pot was moved to a vacuum glove box, and placed for 3 h to collect ball milling materials; step II, 80 g konjac glucomannan, 78 g chitosan and 75 g collagen are taken and added to 245 g water, and stirred for 60 min at a rate of 1200 rim in; half of the ball milling materials was added during die stirring and mixing process at a rate of 20 parts/min; then 55 g ethylene-vinyl acetate copolymer latex powder, 25 g stcaric acid and 12 g ammonium zirconium carbonate were added, mixed and stirred continuously for 30 min at a rotating rate of 1200 r/min; meanwhile, the remaining half of the ball milling materials was added at a rate of 20 parts/min during the stirring and mixing process to obtain a moss adhesive; the adhesive had a tensile shear strength of 10.6 Mpa and a peel strength of 11.8 N/mm.

Example 3:

A method for applying a moss sporophore suspension containing ehitosan/gluean in greenery coverage of a bare land, included the following steps: step I, capsula of Racomitrium japonicum was collected, and the capsula was cut to collect spores when calyptra of the capsula began falling off, distilled water was used to elute spores residual on the capsula; a 3.5(14,w/w hydrogen peroxide solution was added to the spores, and shaken for 8 min to obtain a mixed solution of disinfected spores and hydrogen peroxide; where a volume ratio of the spores to the 3.5%w/w hydrogen peroxide solution was 1:10; step 11, 2 mL mixed solution of disinfected spores and hydrogen peroxide was inoculated onto a 10 mL spore germination-inducing culture medium for spore germination-inducing culture for 40 d to obtain a suspension containing spores and protonema; conditions of the spore germination-inducing culture were as follows: temperature was 25°C, illumination intensity was 28001x, illumination time during the first 1-5 d was 22 h/d, illumination time after the 6thdav was 8 h/d; the moss spore germination-inducing culture medium included as follows: an MS medium + 0.8 mg/L KT (kinetin) + 0.2 mg/L GA3 (gibbcrellin) + 20 g/L white sugar with a pH value of 6.0; step III, 2 mL of the suspension containing spores and protonema in the step II was taken and added to a 10 mL solid subculture medium for subculturing for 30 d to obtain a suspension containing green spore colonies and protonema; the subculturing conditions were as follows: a temperature was 28°C, an illumination intensity was 28001x, an illumination time was 8 h/d; and the solid subculture medium included as follows: an MS medium + 0.8mg/L BA (benzylamino adenine) + 0.2 mg/L 2,4-D(2,4-dichlorphenoxyacetic acid) + 0,3 mg/L TAA (auxin) + 20 g/L white sugar with a pH value of 6.0; step IV, 2 mL of the suspension containing green spore colonies and protonema in the step HT was taken and mixed with a 10 mL fluid medium to obtain a mixed solution; then the mixed solution was subjected to shaking culture for 25 d on a shaker at a rotating rate of 100 r/min to obtain a dark green suspension; the shaking culture conditions on the shaker were as follows: tunperature was 25°C, illumination intensity was 28001x, illumination time was 8 h/d; the dark green suspension was obtained until the mixed solution thickened and turned dark green; and the fluid medium: an MS medium + 0,1 mg/L BA + 0.4mg/L NAA (naphthylacetic acid) + 20 g/L white sugar with a pH value of 6.0; step V, 0.8 g chitosan, 0.4 g glucan, 0.3 g ammonium ferric citrate, 0.2 g phenoxyacetic acid, 0.15 g zinc sulfate and 0.2 g potassium nitrate were added to 10 g dark green suspension in the step IV, and stirred to obtain a moss sporophore suspension; step VI, a moss adhesive was evenly painted onto a bare rock, and the moss sporophore suspension was evenly sprayed on a surface of the moss adhesive; the surface was covered with a sunshading net and subjected to spray irrigation in time, such that a surface layer of the polluted bare land kept a humidity of 23%; after 30 d incubation, the coverage of moss was up to 95%; and after 60 d incubation, it was not found that the moss layer was separated from the rock; a preparation method of the toss adhesive was as follows: 18 g melamine, 15 g urea, 75 g attapulgite and 72 g sepiolite powder were added to a ball null pot; and a milling ball was added to the ball mill pot and liquid nitrogen was fed to the ball mill pot, such that the materials were immersed into the liquid nitrogen; the temperature was -170°C, and the volatile amount and feed amount of the liquid nitrogen were kept balance, so as to stabilize the liquid level; ball milling was performed for 40 min after keeping constant temperature for 15 min; after the ball milling was completed, the ball mill pot was moved to a vacuum glove box, and placed for 3 h to collect ball milling materials; step II, 80 g konjac glucomannan, 7/I g chitosan and 75 g collagen were taken and added to 245 g water, and stirred for 60 min at a rate of 1200 r/mim half of the ball milling materials was added during the stirring and mixing process at a rate of 20 g/min; then 55 g ethylene-vinyl acetate copolymer latex powder, 25 g stearic acid and 12 g ammonium zirconium carbonate were added, mixed and stiffed continuously for 30 min at a rotating rate of 1200 r/min; meanwhile, the remaining half of the ball milling materials was added at a rate of 20 g/min during the stirring and mixing process to obtain a dedicated moss adhesive; the adhesive had a tensile shear strength of 10.6 Mpa, and a peel strength of 11.8 N/mm.

Example 4:

A method for applying a moss sporophore suspension containing chitosan/glucan in greenery coverage of a bare land, included the following steps: step I, capsula of Racomitrium japonicum was collected, and the capsula was cut to collect spores when calyptra of the capsula began falling off, distilled water was used to elute spores residual on the capsula; a 3.5%wiw hydrogen peroxide solution was added to the spores, and shaken for 8 min to obtain a mixed solution of disinfected spores and hydrogen peroxide; where a volume ratio of the spores to the 3.5%w/w hydrogen peroxide solution was 1.10; step 11, 2 mL mixed solution of disinfected spores and hydrogen peroxide was inoculated onto a 10 mL spore gennination-inducing culture medium for spore gennination-inducing culture for 40 d to obtain a suspension containing spores and protonema, conditions of the spore gennination-inducing culture were as follows: temperature was 25°C, illumination intensity was 28001x, illumination Lime during die first 1-5 d was 22 h/d, illumination time after die 6th day was 8 hid; the moss spore germination-inducing culture medium included as follows: an MS medium + 0.8mg/L KT (kinetin) + 0.2 mg/L GA3 (gibberellin) + 20 g/L white sugar with a pH value of 6.0; step HT, 2 mL of the suspension containing spores and protonema in the step IT was taken and added to a 10 mL solid subculture medium for subculturing for 30 d to obtain a suspension containing green spore colonies and protonema; the subculturing conditions were as follows: a temperature was 28°C, an illumination intensity was 28001x, an illumination time was 8 hid; and the solid subculture medium included as follows: an MS medium + 0.8mg/L BA (benzylamino adenine) + 0.2 mg/L 2,4-D(2,4-dichlorphenoxyacetic acid) + 0.3 mg/L IAA (auxin) + 20 g/L white sugar with a pH value of 6.0; step IV, 2 mL of the suspension containing green spore colonies and protonema in the step III was taken and mixed with a 10 mL fluid medium to obtain a mixed solution; then the mixed solution was subjected to shaking culture for 25 d on a shaker at a rotating rate of 100 r/min to obtain a dark green suspension; the shaking culture conditions on the shaker were as follows: temperature was 25°C, illumination intensity was 28001x, illumination time was 8 hid; the dark green suspension was obtained until the mixed solution thickened and turned dark green; and the fluid medium: an MS medium + 0.1 mg/L BA + 0.4mg/L NAA (naphthylacetic acid) + 20 g/L white sugar with a pH value of 6.0; step V.0.8 g chitosan, 0.4 g glucan, 0.3 g ammonium ferric citrate, 0.2 g phenoxy acetic acid, 0.15 g zinc sulfate and 0.2 g potassium nitrate were added to 10 g dark green suspension in the step IV, and stirred to obtain a moss sporophore suspension; step VI, a moss adhesive was evenly painted onto a bare rock, and the moss sporophore suspension was evenly sprayed on a surface of the moss adhesive; the surface was covered with a sunshading net and subjected to spray irrigation in time such that a surface laver of the polluted bare land kept a humidity of 23%; after 30 d incubation, the coverage of moss was up to 99%; and after 60 d incubation, it was not found that the moss layer was separated from the rock; a preparation method of the toss adhesive was as follows: 18 g melamine, 15 g urea, 75 g attapulgite and 72 g sepiolite powder were added to a ball mill pot; and a milling ball was added to the ball mill pot and liquid nitrogen was fed to the ball mill pot, such that the materials were immersed into the liquid nitrogen; the temperature was -170°C, and the volatile amount and feed amount of the liquid nitrogen were kept balance, so as to stabilize the liquid level; ball milling was performed for 40 min after keeping constant temperature for 15 min; after the ball milling was completed, the ball mill pot was moved to a vacuum glove box, and placed for 3 h to collect ball milling materials; step II, 80 g konjac glucomannan, 78 g chitosan and 75 g modified collagen were taken and added to 245 g water, and stirred for 60 mm at a rate of 1200 r/min; half of the ball milling materials was added during the stirring and mixing process at a rate of 20 g/min; then 55 g ethylene-vinyl acetate copolymer latex powder, 25 g stearic acid and 12 g ammonium zirconium carbonate were added, mixed and stirred continuously for 30 min at a rotating rate of 1200 r/min; meanwhile, the remaining half of the ball milling materials was added at a rate of 20 g/min during the stirring and mixing process to obtain a moss adhesive; a preparation method of the modified collagen was as follows: 80 g collagen were added to 1000 g water for ultrasonic treatment for 45 min: 8 g eatechinie acid were added and then ultrasonic treatment was performed for 30 min; then 20 g poly L-glutamic acid were added; and then the mixed materials were heated up to 60°C, and irradiated by UV light, stirred for reaction for 30 min, and subjected to centrifugal separation, thus obtaining the modified collagen; the ultrasonic treatment had a power of 120 W and a frequency of 50 kHz; the UV light had a irradiation power of 300 W, and the UV light had a wavelength of 254 nm. By the modification of collagen, catechinic acid and poly L-glutamic acid were reacted with collagen to form the intramolecular and intermolecular crosslinking. These crosslink bonds intertwined with each other to form a compact net structure, which greatly improved intermolecular cohesion, thus enhancing the strength of the adhesive. The dedicated moss adhesive prepared in the example had a tensile shear strength of 12.7 Mpa, and a peel strength of 13.5 N/mm.

Example 5:

A method for applying a moss sporophore suspension containing chitosan/glucan in greenery coverage of a bare land, included the following steps: step 1, capsula of Racomitrium japonicum was collected, and the capsula was cut to collect spores when calyptra of the capsula began falling off, distilled water was used to elute spores residual on the capsula; a 3.5%w/w hydrogen peroxide solution was added to the spores, and shaken for 8 min to obtain a mixed solution of disinfected spores and hydrogen peroxide; where a volume ratio of the spores to the 3.5%w/w hydrogen peroxide solution was 1:10; step II, 2 mL mixed solution of disinfected spores and hydrogen peroxide was inoculated onto a 10 mL sporc germination-inducing culture medium for spore germination-inducing culture for 40 d to obtain a suspcnsion containing spores and protonema; conditions of the spore germination-inducing culture were as follows: temperature was 25°C, illumination intensity was 28001x, illumination time during the first 1-5 d was 22 h/d, illumination time after the 61h day was 8 11/d; the moss spore gemination-inducing culture medium included as follows: an MS medium -1 0.8mg/L KT (kinetin) + 0.2 mg/L GA3 (gibberellin) + 20 g/L white sugar with a pH value of 6.0; step 111, 2 mL of the suspension containing spores and protonema in the step IT was taken and added to a 10 mL solid subculture medium for subculturing for 30 d to obtain a suspension containing green spore colonies and protonema; the subculturing conditions were as follows: a temperature was 28°C, an illumination intensity was 28001x, an illumination time was 8 h/d; and the solid subculture medium included as follows: an MS medium + 0.8mg/L BA (benzylamino adenine) + 0.2 mg/L 2,4-D(2,4-dichloiphenoxyacetic acid) + 0.3 mg/L IAA (auxin) + 20 g/L white sugar with a pH value of 6.0; step IV, 2 mL of the suspension containing green spore colonies and protonema in the step 111 was taken and mixed with a 10 mL fluid medium to obtain a mixed solution; then the mixed solution was subjected to shaking culture for 25 d on a shaker at a rotating rate of 100 r/min to obtain a dark green suspension; the shaking culture conditions on the shaker were as follows: temperature was 25°C, illumination intensity was 2800Ix, illumination time was 8 h/d, the dark green suspension was obtained until the mixed solution thickened and turned dark green; and the fluid medium: an MS medium + 0.1 mg/L BA + 0.4mg/L NAA (naphthylacctic acid) + 20 g/L white sugar with a pH value of 6.0; step V, 0.8 g chitosan, 0.4 g glucan, 013 g ammonium ferric citrate, 02 g phenoxyacetic acid, 0.15 g zinc sulfate and 0.2 g potassium nitrate were added to 10 g dark green suspension in the step IV, and stirred to obtain a moss sporophore suspension; step VI, a moss adhesive was evenly painted onto a bare soil polluted by heavy metal (the soil was subjected to impurity removal, dried, ground, treated by wet ashing, and filtered; then filtrate was dried, then dissolved by adding distilled water, and tested by Atomic Absorption Spectrometry; where lead content in the soil was 45.455 mg/kg); and the moss sporophore suspension was evenly sprayed on a surface of the moss adhesive; the surface was covered with a sunshading net and subjected to spray irrigation in time, such that a surface layer of the polluted bare soil kept a humidity of 23%; after 30 d incubation, the coverage of moss was up to 92%; after being cultured for 90 d, the moss was collected, cleaned, dried, ground, treated by wet ashing, and filtered; then filtrate was dried, then dissolved by adding distilled water, and tested by Atomic Absorption Spectrometry; lead content in the soil was 11.625 mg/kg through test.

a preparation method of the toss adhesive was as follows: 18 g melamine, 15 g urea, 75 g attapulgite and 72 g sepiolite powder were added to a ball mill pot; and a milling ball was added to the ball mill pot and liquid nitrogen was fed to the ball mill pot, such that the materials were immersed into the liquid nitrogen; the temperature was -170°C, and the volatile amount and feed amount of the liquid nitrogen were kept balance, so as to stabilize the liquid level; ball milling was performed for 40 min after keeping constant temperature for 15 min; after the ball milling was completed, the ball mill pot was moved to a vacuum glove box, and placed for 3 h to collect ball milling materials; step II, 80 g konjac glucomannan, 78 g chitosan and 75 g collagen were taken and added to 245 g water, and stirred for 60 min at a rate of 1200 r/min; half of the ball milling materials was added during the stirring and mixing process at a rate of 20 g/min; then 55 g ethylene-vinyl acetate copolymer latex powder, 25 g stcaric acid and 12 g ammonium zirconium carbonate were added, mixed and stiffed continuously for 30 min at a rotating rate of 1200 r/min; meanwhile, the remaining half of the ball milling materials was added at a rate of 20 g/min during the stirring and mixing process to obtain a moss adhesive; the adhesive had a tensile shear strength of 10.6 Mpa, and a peel strength of 11.8 N/mm,

Example 6:

A method for applying a moss sporophore suspension containing chitosan/glucan in greenery coverage of a bare land, included the following steps: step I, capsula of Racomitrium japonicum was collected, and the capsula was cut to collect spores when calyptra of the capsula began falling off, distilled water was used to elute spores residual on the capsula; a 3.5%w/w hydrogen peroxide solution was added to the spores, and shaken for 8 min to obtain a mixed solution of disinfected spores and hydrogen peroxide; where a volume ratio of the spores to the 3.5%w/w hydrogen peroxide solution was 1:10; step II, 2 mL mixed solution of disinfected spores and hydrogen peroxide was inoculated onto a 10 mL sporc germination-inducing culture medium for spore germination-inducing culture for 40 d to obtain a suspension containing spores and protonerna; conditions of the spore germination-inducing culture were as follows: temperature was 25°C, illumination intensity was 28001x, illumination time during the first 1-5 d was 22 hid, illumination time after the 6111 day was 8 hid; the moss spore germination-inducing culture medium included as follows: an MS medium -1 0.8mg/L KT (kinetin) + 0.2 mg/L GA3 (gibberellin) + 20 g/L white sugar with a pH value of 6.0: step III, 2 mL of the suspension containing spores and protonema in the step II was taken and added to a 10 mL solid subculture medium for subculturing for 30 d to obtain a suspension containing green spore colonies and protonema; the subculturing conditions were as follows: a temperature was 28°C, an illumination intensity was 28001x, an illumination time was 8 hid; and the solid subculture medium included as follows: an MS medium + 0.8mg/L BA (bcnzylamino adenine) + 0.2 mg/L 2,4-D(2,4-dichlorphenoxyacetic acid) + 0.3 mg/L TAA (auxin) + 20 g/L white sugar with a pH value of 6.0; step IV, 2 mL of the suspension containing green spore colonies and protonema in the step 111 was taken and mixed with a 10 mL fluid medium to obtain a mixed solution; then the mixed solution was subjected to shaking culture for 25 d on a shaker at a rotating rate of 100 r/min to obtain a dark green suspension; the shaking culture conditions on the shaker were as follows: temperature was 25°C, illumination intensity was 28001x, illumination time was 8 hid; the dark green suspension was obtained until the mixed solution thickened and turned dark green; and the fluid medium: an MS medium + 0.1 mg/L BA + 0.4mg/L NAA (naphthylacetic acid) + 20 g/L white sugar with a pH value of 6.0; step V, 0.8 g chitosan, 0.4 g glucan, 0.3 g ammonium ferric citrate, 0.2 g phenoxyacetic acid, 0.15 g zinc sulfate and 0.2 g potassium nitrate were added to 10 g dark green suspension in the step IV, and stirred to obtain a moss sporophore suspension; step VI, a moss adhesive was evenly painted onto a bare soil polluted by heavy metal (the soil was subjected to impurity removal, dried, ground, treated by wet ashing, and filtered; then filtrate was dried, then dissolved by adding distilled water, and tested by Atomic Absorption Spectrometry; where lead content in the soil was 45.455 mg/kg), and the moss sporophore suspension was evenly sprayed on a surface of the moss adhesive; the surface was covered with a sunshading net and subjected to spray irrigation in time, such that a surface layer of the polluted bare soil kept a humidity of 23%; after 30 d incubation, the coverage of moss was up to 96%; the moss after being cultured for 90 d was collected, cleaned, dried, ground, treated by wet ashing, and filtered; then filtrate was dried, and dissolved by adding distilled water, and tested by Atomic Absorption Spectrometry; after test, lead content in the soil was I 5.854mg/kg.

a preparation method of the toss adhesive was as follows: 18 g melamine, 15 g urea, 75 g attapulgite and 72 g sepiolite powder were added to a ball mill pot; and a milling ball was added to the ball mill pot and liquid nitrogen was fed to the ball mill pot, such that the materials were immersed into the liquid nitrogen; the temperature was -170°C, and the volatile amount and feed amount of the liquid nitrogen were kept balance, so as to stabilize the liquid level; ball milling was performed for 40 min after keeping constant temperature for 15 min; after the ball milling was completed, the ball mill pot was moved to a vacuum glove box, and placed for 3 h to collect ball milling materials; step II, 80 g konjac glucomannan, 78 g chitosan and 75 g modified collagen were taken and added to 245 g water, and stirred for 60 mm at a rate of 1200 r/min; half of the ball milling materials was added during the stirring and mixing process at a rate of 20 g/min; then 55 g ethylene-vinyl acetate copolymer latex powder, 25 g stearic acid and 12 g ammonium zirconium carbonate were added, mixed and stirred continuously for 30 min at a rotating rate of 1200 r/min; meanwhile, the remaining half of the ball milling materials was added at a rate of 20 g/min during the stirring and mixing process to obtain a moss adhesive; a preparation method of the modified collagen was as follows: 80 g collagen were added to 1000 g water for ultrasonic treatment for 45 min: 8 g catechinic acid were added and then ultrasonic treatment was performed for 30 min; then 20 g poly L-glutamic acid were added; and then the mixed materials were heated up to 60°C, and irradiated by UV light, stirred for reaction for 30 min, and subjected to centrifugal separation, thus obtaining the modified collagen; the ultrasonic treatment had a power of 120 W and a frequency of 50 kHz; the UV light had a irradiation power of 300 W, and the UV light had a wavelength of 254 nm. By the modification of collagen, catechinic acid and poly L-glutamic acid were reacted with collagen to form the intramolecular and intermolecular crosslinking. These crosslink bonds intertwined with each other to form a compact net structure, which greatly improved intermolecular cohesion, thus enhancing the strength of the adhesive. The dedicated moss adhesive prepared in the example had a tensile shear strength of 12.7 Mpa, and a peel strength of 13.5 N/mm.

Even though the embodiments of the present invention have been disclosed as the above, the present invention is not merely limited to the applications listed in the description and embodiments thereof, but should be completely applicable to the various technical fields suitable for the present invention. Moreover, a person skilled in the art can easily achieve additional amendments. Therefore, the present invention is not limited to specific details and examples described herein within the general concept defined by the claims and the equivalent protection scope.

Claims

Claims I. A method for applying a moss sporophore suspension containing chitosan/glucan in greenery coverage of a bare land, characterized by comprising the following steps: step I, collecting capsula of moss, and cutting the capsula to collect spores when calyptra of the capsula begins falling off, using distilled water to elute spores residual on the capsula; adding a 3-4%w/w hydrogen peroxide solution to the spores, and shaking for 6-10 mm to obtain a mixed solution of disinfected spores and hydrogen peroxide; wherein a volume ratio of the spores to the 3-4%w/w hydrogen peroxide solution is I: 10; step 11, inoculating the mixed solution of disinfected spores and hydrogen peroxide onto a spore germination-inducing culture medium for spore germination-inducing culture for 30-50 d to obtain a suspension containing endospores and protonema; step TIT, taking arid adding the suspension containing endospores and protonema onto a solid subculture medium for subculturing for 28-32 d; thus obtaining a suspension containing green endospore colonies and protonema.step IV, taking the suspension containing green endospore colonies and protonema in the step III and mixing with a fluid medium to obtain a mixed solution, then subjecting the mixed solution to shaking culture for 25-30 d, thus obtaining a dark green suspension; step V, adding chitosan, glucan, ammonium ferric citrate. phenoxyacetic acid, zinc sulfate and potassium nitrate to the dark green suspension in the step TV, and stirring them to obtain a moss sporophore suspension; step VI, evenly painting a moss adhesive onto a bare land, and evenly spraying the moss sporophore suspension on a surface of the moss adhesive; covering the surface with a sunshading net and performing spray irrigation in time, such that a surface layer of the polluted bare land keeps a humidity within 20-25%; a preparation method of the toss adhesive is as follows: according to parts by weight, 10-30 parts of melamine, 10-20 parts of urea, 60-90 parts of attapulgite and 50-80 parts of sepiolite powder are added to a ball mill pot; and a milling ball is added to the ball mill pot and liquid nitrogen is fed to the ball mill pot, such that the materials are immersed into the liquid nitrogen at a temperature within -160°C to -175°C, and the volatile amount and feed amount of the liquid nitrogen are kept balance, so as to stabilize the liquid level; ball milling is performed for 30-45 min after keeping constant temperature for 15 min; after the ball milling is completed, the ball mill pot is moved to a vacuum glove box, and placed for 3-5 h to collect ball milling materials; step II, according to parts by weight, 50-100 parts of konjac glucomannan, 50-100 parts of chitosan and 50-100 parts of modified collagen are taken and added to 200-300 parts of water, and stirred for 30-60 min at a rate of 1000-1200 r/min; half of the ball milling materials is added during the stirring and mixing process at a rate of 10-30 parts/min; then 40-60 parts of latex powder, 10-30 parts of stearic acid and 5-15 parts of ammonium zirconium carbonate are added, mixed and stirred continuously for 30-45 min at a rotating rate of 1000-1200 r/min, meanwhile, the remaining half of the ball milling materials is added at a rate of 10-30 parts/min during the stirring and mixing process to obtain a dedicated moss adhesive; a preparation method of the modified collagen is as follows: according to parts by weight, 50-100 parts of collagen are added to 1000-1200 parts of water for ultrasonic treatment for 30-45 min; 5-8 parts of catechinic acid is added and then ultrasonic treatment is performed for 15-30 min; then IS-25 parts of poly L-glutamic acid are added; and then the mixed materials are heated up to 45-60°C, and irradiated by UV light, stirred for reaction for 15-30 min, and subjected to centrifugal separation, thus obtaining the modified collagen.
2. The method for applying a moss sporophore suspension comprising chitosan/glucan in greenery coverage of a bare land of claim 1, characterized in that in the step 11, conditions of the spore gemination-inducing culture are as follows: temperature is 22-28°C, illumination intensity is 25003000k. illumination time during the first 1-5 d is 20-24 h/d, illumination time after the 6th day is 8 hid; the moss spore germination-inducing culture medium comprises as follows: MS + 0.5-1.0 mg/L KT + 0.2 mg/L 0A3 + 20 g/L white sugar with a pH value of 6.0; and a volume ratio of the mixed solution of disinfected spores and hydrogen peroxide to the spore gennination-inducing culture medium is 1:5.
3. The method for applying a moss sporophore suspension comprising chitosan/glucan in greenery coverage of a bare land of claim 1, characterized in that in the step III, the subculturing conditions are as follows: a temperature is 22-28°C, an illumination intensity is 2500-30001x, an illumination time is 8 h/d; and the solid subculture medium comprises as follows: MS + 0.5-1.0 mg/LBA 0.2 mg/L 2, 4-D + 0,3 mg/L IAA + 20 g/L white sugar with a pH value of 6.0.
4. The method for applying a moss sporophore suspension comprising chitosan/glucan in greenery coverage of a bare land of claim 1, characterized in that in the step IV, the shaking culture is performed on a shaker at 100 rimin; the shaking culture conditions on the shaker are as follows: temperature is 22-28°C, illumination intensity is 2500-30001x, illumination time is 8 h/d; a dark green suspension is obtained until the mixed solution thickens and turns dark green; and the fluid medium: MS + 0.1mg/L BA + 02-0.5 mg/L NAA + 20 g/L white sugar with a pH value of 6.0.
5. The method for applying a moss sporophorc suspension comprising chitosan/glucan in greenery coverage of a bare land of claim 1, characterized in that in the step V, according to parts by weight, 0.5-1 parts of chitosan, 0.3-0.5 parts of glucan, 0.2-0.4 parts of ammonium ferric citrate, 0.1-0.3 parts of phenoxyacetic acid, 0.1-02 parts of zinc sulfate and 0.1-0.3 parts of potassium nitrate are added to 100 parts of the dark green suspension in the step IV; and then stirring is performed to obtain a moss sporophorc suspension.
6. The method for applying a moss sporophore suspension comprising chitosan/glucan in greenery coverage of a bare land of claim 1, characterized in that the bare land is any one of a bare rock, a bare ground, a bare sandy land, a bare project profile, a bare land polluted by heavy metals, a bare land polluted by pesticides, and a radioactive source-polluted soil.
7. The method for applying a moss sporophore suspension comprising chitosan/glucan in greenery coverage of a bare land of claim 1, characterized in that the latex powder is one or more than two of ethylene-vinyl acetate copolymer latex powder, acrylate-styrene copolymerized rubber powder, ternary-copolymerized rubber powder of vinyl acetate, acrylatc and higher fatty acid vinyl ester.
8. The method for applying a moss sporophore suspension comprising chitosan/glucan in greenery coverage of a bare land of claim 1, characterized in that the ultrasonic treatment has a power of 80120 W and a frequency of 30-50 kHz; and the UV light has an irradiation power of 50-500W and the UV light has a wavelength of 220-400 nm.