CN115299329A - Method for culturing odontopathy and application thereof - Google Patents
Method for culturing odontopathy and application thereof Download PDFInfo
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- CN115299329A CN115299329A CN202210930921.2A CN202210930921A CN115299329A CN 115299329 A CN115299329 A CN 115299329A CN 202210930921 A CN202210930921 A CN 202210930921A CN 115299329 A CN115299329 A CN 115299329A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
- A01G24/12—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material containing soil minerals
- A01G24/15—Calcined rock, e.g. perlite, vermiculite or clay aggregates
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
- A01G24/28—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material containing peat, moss or sphagnum
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05B—PHOSPHATIC FERTILISERS
- C05B7/00—Fertilisers based essentially on alkali or ammonium orthophosphates
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G5/00—Fertilisers characterised by their form
- C05G5/20—Liquid fertilisers
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- Chemical & Material Sciences (AREA)
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- Organic Chemistry (AREA)
- Soil Sciences (AREA)
- Inorganic Chemistry (AREA)
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- Cultivation Of Plants (AREA)
Abstract
The invention provides a method for culturing odontopathy and application thereof, relating to the technical field of environmental engineering. The invention provides a culture method of dentritic moss, peat soil and vermiculite are added into growth nutrient soil, which is beneficial to the growth of the dentritic moss, and the added nutrient solution provides elements required during the growth of the dentritic moss, so that the growth speed of the dentritic moss is improved. Experiments prove that the cultivation method for the sphagnum dentiger can obviously improve the growth speed of the sphagnum dentiger and the photosynthesis capability of the sphagnum dentiger when the sphagnum dentiger is planted in a slope in a large area, is easy to realize the large-area planting of the sphagnum dentiger and has good application prospect for repairing the good ecological environment of a mining area.
Description
Technical Field
The invention relates to the technical field of environmental engineering, in particular to a method for culturing odontopathy and application thereof.
Background
The biological soil crust layer mainly comprises blue algae, green algae, bacteria, fungi, lichen and moss. Mosses are the predominant component in the fixation of dune cortex, a biological group with the largest biomass in the cortex, and play a major ecological role in the annual thickening, maintenance, nutrient accumulation and water dynamics of the cortex. The soil-borne ratchets moss is widely distributed, mainly distributed in arid areas, semi-arid areas and alpine ecosystems, and can also live in environments with high temperature, high light intensity, unbalanced organic nutrition and even extreme drought. Researches show that the soil growth is an important component of biological soil crust with the odontopathy, algae, lichens and soil microorganisms, plays unique advantages in aspects of land reclamation, field planting, reclamation, soil environment improvement, high-cold grassland recovery promotion and the like, makes important contribution to ecological function balance of arid areas, is a key group of key fragile ecological areas such as a windy and sandy area, a northwest arid area, a loess plateau area, a Qinghai-Tibet plateau area and the like in the north of China, and is also one of important indicator plants of climate change and the fragile degree of a living environment of the Qinghai-Tibet plateau.
Moreover, coal resource exploitation and development bring about a series of environmental problems, water and soil in a mining area are not well protected, sand and dust are scattered, bare and loose accumulations are easy to be washed by rainwater, vegetation is rare, the ecological environment is fragile, and problems of the ecological environment emerge endlessly in arid and semiarid areas of a coal mine, so that the aim of restoring the ecological environment in the coal mining area to a certain extent is achieved by planting the sphagnum in a large area. However, although there are artificial culture methods for other bryophytes, there are still a lack of methods for large-area planting of bryophytes at present due to the significant differences in the growth characteristics and adaptability to the environment of different bryophytes, which seriously hinders the cultivation and application of bryophytes.
Disclosure of Invention
In view of the above, the present invention is directed to provide a cultivation method of odontopathy, which can improve the growth rate of odontopathy and realize large-area cultivation of odontopathy.
In order to solve the technical problems, the invention provides the following technical scheme:
the invention provides a culture method of odontopathy, which comprises the step of placing pretreated odontopathy on the surface of nutrient soil added with a matrix for culture, wherein the matrix is a mixture of peat soil and vermiculite.
Preferably, the mass ratio of the peat soil to the vermiculite is 1-3:1-3.
Preferably, the rhodobryum dentium is pulverized rhodobryum dentium or separated stem and leaf rhodobryum dentium.
Preferably, the pretreatment comprises 75% alcohol treatment and sterile water cleaning in sequence, and the time of the 75% alcohol treatment is 15-45 s.
Preferably, hormones are sprayed during the culturing process.
Preferably, the hormone is one or more of 0.01 μ M6-BA, 0.1mg/L NAA, 1mg/L NAA, 0.01 mg/L2,4-D, 0.01mg/L IAA, 0.01mol/L IBA.
Preferably, the nutrient soil is also added with a Knop nutrient solution; the Knop nutrient solution comprises 1g of calcium nitrate, 0.25g of potassium nitrate, 0.25g of monopotassium phosphate, 0.25g of magnesium sulfate, 0.003g of zinc sulfate and 1000mg of distilled water.
Preferably, the culture is a membrane culture.
Preferably, the temperature of the culture is 21-25 ℃, and the humidity of the culture is 50-55%.
The invention also provides application of the culture method for the sphagnum dentiger in restoring the ecological environment of the mining area.
The invention has the beneficial effects that:
the invention provides a culture method for odontopathy, peat soil and vermiculite are added into growth nutrient soil, growth of the odontopathy is facilitated, the added nutrient solution provides elements required during growth of the odontopathy, and the growth speed of the odontopathy is improved. Experiments prove that the cultivation method for the sphagnum dentiger can obviously improve the growth speed of the sphagnum dentiger and the photosynthesis capability of the sphagnum dentiger when the sphagnum dentiger is planted in a slope in a large area, is easy to realize the large-area planting of the sphagnum dentiger and has good application prospect for repairing the good ecological environment of a mining area.
Drawings
FIG. 1 shows the coverage of the lichen brachypomus under each treatment.
FIG. 2 shows the density of Brachypodium versus dentosis for each treatment.
Detailed Description
The invention provides a method for culturing odontopathy, which comprises the step of placing pretreated odontopathy on the surface of nutrient soil added with a matrix for culturing, wherein the matrix is a mixture of peat soil and vermiculite.
In the invention, the mass ratio of the peat soil to the vermiculite is preferably 1-3:1-3, more preferably 2:1; the thickness of the substrate is preferably > 3cm. The matrix added into the nutrient soil is beneficial to the growth of the odontopathy, can improve the coverage and density of the odontopathy and promote the rapid propagation of the odontopathy. In the invention, the para-odontopathy is fresh and well-grown para-odontopathy, and preferably is crushed para-odontopathy or stem-leaf separation para-odontopathy; the crushed rhodobryum dentium is obtained by crushing overground stem and leaf parts of the rhodobryum dentium. The specific way of pulverization is not particularly limited in the present invention, and the pulverization method conventional in the art may be adopted, and in the specific embodiment of the present invention, the pulverization is preferably pulverization by a pulverizer.
In the invention, the pretreatment comprises 75% alcohol treatment and sterile water cleaning in sequence, and the time of the 75% alcohol treatment is preferably 15s-45s, and more preferably 30s. In the invention, hormone is sprayed in the culture process; the hormone is preferably one or more of 0.01 mu M6-BA, 0.1mg/L NAA, 1mg/L NAA, 0.01 mg/L2, 4-D, 0.01mg/L IAA and 0.01mol/L IBA. The hormone of the invention has promotion effect on protonema differentiation and branch elongation of moss, and is beneficial to large-area culture of the moss. In the invention, the nutrient soil is also added with Knop nutrient solution; the Knop nutrient solution comprises 1g of calcium nitrate, 0.25g of potassium nitrate, 0.25g of monopotassium phosphate, 0.25g of magnesium sulfate, 0.003g of zinc sulfate and 1000mg of distilled water. In the present invention, the addition amount of the Knop nutrient solution is preferably 10% to 70%, more preferably 20%, per bottle based on 60% of the saturated water absorption of each substrate. The Knop nutrient solution can provide elements required by the growth period of the dentures and is beneficial to the rapid propagation of the dentures.
In the present invention, the culture is preferably a membrane culture, specifically: during the cultivation, a film-covering treatment is performed using a plastic film, the purpose of which is to allow the plant growth to be in an environment in which temperature, air humidity and moisture are stable. In the present invention, the temperature of the culture is preferably 21 ℃ to 25 ℃, more preferably 23 ℃; the humidity of the culture is preferably 50% -55%, more preferably 53.2%; the time period of the culture is preferably 30 to 60 days, more preferably 45 days. In the present invention, it is preferable that ventilation is required during the period of high temperature during the cultivation period to prevent the death of the moss due to the high temperature. During the cultivation period according to the present invention, watering is preferably performed by using a water spraying device in the morning and evening of each day. In the present invention, the light intensity during the culture is preferably 3000 to 3500lx, and more preferably 3300xl.
The invention also provides application of the culture method for the sphagnum dentiger in restoring the ecological environment of the mining area.
The present invention will be described in detail with reference to examples for better understanding the objects, technical solutions and advantages of the present invention, but they should not be construed as limiting the scope of the present invention.
In the following examples, unless otherwise specified, all methods are conventional.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Example 1
1) Treating fresh and well-grown overground stem and leaf parts of the denticulate moss with 75% alcohol for 30s, cleaning with sterile water for 3 times until residual disinfectant is cleaned, absorbing surface water with filter paper, and pulverizing with a pulverizer for later use;
2) Taking the nutrient soil by using a rectangular plastic box, and then mixing peat soil into the nutrient soil: vermiculite =2:1, then uniformly scattering the smashed sphagnum sample on the surface of nutrient soil for film-covering culture for 7 days, spraying 0.01 mu M6-BA in the culture process, setting the illumination intensity at 3300lx during culture, placing a thermometer in the film to control the temperature at 23 ℃, and setting a humidity detector to control the humidity at 53.2%. Meanwhile, a Knop nutrient solution prepared from 1g of calcium nitrate, 0.25g of potassium nitrate, 0.25g of monopotassium phosphate, 0.25g of magnesium sulfate, 0.003g of zinc sulfate and 1000mg of distilled water was added to the nutrient soil.
Example 2
1) Treating fresh and well-grown overground stem and leaf parts of the sphagnum by using 75% alcohol for 30s, cleaning the overground stem and leaf parts of the sphagnum by using sterile water for 3 times until residual disinfectant is cleaned, absorbing surface water by using filter paper, and crushing the ground overground stem and leaf parts by using a crusher for standby;
2) Taking the nutrient soil by using a rectangular plastic box, and then mixing peat soil: vermiculite =2:1, uniformly placing the entire plant of the sphagnum moss sample on the surface of nutrient soil for film-covering culture for 7 days, spraying 0.01mg/L IAA in the culture process, setting the illumination intensity at 3000lx during culture, placing a thermometer in a membrane to control the temperature at 21 ℃, and setting a humidity detector to control the humidity at 50%. Meanwhile, a Knop nutrient solution prepared from 1g of calcium nitrate, 0.25g of potassium nitrate, 0.25g of monopotassium phosphate, 0.25g of magnesium sulfate, 0.003g of zinc sulfate and 1000mg of distilled water was added to the nutrient soil.
Example 3
1) Treating fresh and well-grown overground stem and leaf parts of the sphagnum by using 75% alcohol for 30s, cleaning for 3 times by using sterile water until residual disinfectant is cleaned, then absorbing surface water by using filter paper, separating stem and leaf of the sphagnum, and mixing the separated stem and leaf of the sphagnum for later use;
2) Taking the nutrient soil by using a rectangular plastic box, and then mixing peat soil into the nutrient soil: vermiculite =2:1, uniformly scattering the smashed sphagnum sample on the surface of nutrient soil for film-covering culture for 7 days, spraying 1mg/L NAA in the culture process, setting the illumination intensity at 3500lx during culture, placing a thermometer in the film to control the temperature at 25 ℃, and setting a humidity detector to control the humidity at about 55%. Meanwhile, a Knop nutrient solution prepared from 1g of calcium nitrate, 0.25g of potassium nitrate, 0.25g of monopotassium phosphate, 0.25g of magnesium sulfate, 0.003g of zinc sulfate and 1000mg of distilled water was added to the nutrient soil.
Example 4
Effect of different growth substrates on moss growth
1) Grouping
The test was conducted by setting 16 treatment groups and 1 control group, and dividing the test into three major parts, i.e., different substrate mixtures, different layer substrates, and blank controls, wherein the mixing ratio of the substrates in each group is shown in table 1 below.
TABLE 1 different groups of matrix additions
2) Measurement index and method
Coverage degree: the 25-point sampling method was used. Dividing a self-made square sample prescription with the side length of 5cm into 25 small squares with the side length of 1cm, counting the number of moss in a fixed point in the 25 small squares, measuring 6 sample prescriptions in each culture bottle, and calculating the coverage degree of the moss according to the percentage of the moss to the total number of the investigated points, wherein the result is shown in figure 1.
Density: a fixed sample method is adopted. The small square sample of 1cm is placed in a culture bottle for 10 times, the number of the moss plants on each small square sample is counted, the average value is taken as the density value, and the result is shown in figure 2.
Chlorophyll content: on day 90, fresh weight was sampled and weighed, and then cut into pieces and mixed uniformly, 0.1g of the mixture was weighed out and used for measuring the chlorophyll content of the tooth moss sample, and the chlorophyll content was measured by a 95% ethanol extraction colorimetric method, and the results are shown in table 2.
TABLE 2 chlorophyll content variation in dentures with different substrate treatments
Remarking: different lower case letters in the same column indicate that the same combination differed significantly at the 0.05 level, with treatments 7, 8, 9, 10, 13, 14, 15, 16 not growing.
And (3) determining the physicochemical property of the matrix: the water content (MC) of the matrix was measured by a weighing method, the pH (pH) was measured by a glass electrode method, the particle size was measured by a laser particle sizer, and classified into gravel (> 1 mm), sand (0.05-1 mm), powder (0.001-0.05 mm) and sticky particles (< 0.001 mm) according to california, and the measurement results are shown in table 3.
TABLE 3 analysis of physicochemical properties of substrates at different ratios
3) Analysis of results
As can be seen from FIGS. 1 and 2, treatment 3 had the greatest coverage and density of dentures in the combination of treatments 1-10 with different base mixes, with 70.67% coverage and a density of 12.2 strain cm -2 Significantly higher than treatment 17 (CK) and other treatments, best for dental moss growth; after 1 treatment, the cover degree was 46%, and the density was 7.09 strain/cm -2 . In the treatment combination of 11-16 with different layer compositions, the coverage and density of the treatment 11 to the dentures are the maximum, the coverage is 34 percent, and the density is 6.34 strains per cm -2 The growth condition is better; and the rhodobryum dentium has no growth in the matrix consisting of perlite, vermiculite or the mixture of the two in proportion. The single perlite and vermiculite are adopted as the substrates, so that the growth of the sphagnum leptospira is not facilitated, and the requirement of the nutrients in the growth process of the sphagnum leptospira cannot be met probably because the nutrients in the perlite and the vermiculite are single. Peat soil: vermiculite =2:1,20% of Dictyotaceae under treatment with knop nutrient solutionThe growth is best.
As can be seen from the results in table 2, treatment 3 had the highest chlorophyll a content, significantly higher than each of the other treatments (P < 0.05), followed by treatment 1, treatment 6 having the lowest chlorophyll a content; the chlorophyll b content and the carotenoid content have no obvious difference among the treatments, and in the chlorophyll b content, the chlorophyll b content of treatment 5 is the largest, the chlorophyll b content of treatment 17 (CK) is the smallest, and the carotenoid content of treatment 11 is the largest; the total chlorophyll content of treatment 3 was the highest. The total content of chlorophyll is ordered as: treatment 3> treatment 5> treatment 17> treatment 2> treatment 4> treatment 1> treatment 11> treatment 12> treatment 6, which shows that different culture media have a significant effect on the chlorophyll content of the sphagnum dentiger, wherein treatment 3 can significantly increase the chlorophyll content in leaves and significantly increase the photosynthetic capacity of the sphagnum dentiger.
As can be seen from the results in Table 3, there was a difference in the physicochemical properties of all the substrates. The pH varied from 6.97 to 7.47, with treatment 3 substrate pH at a minimum of 6.97, significantly lower than other treatments (P < 0.05). Treatment 3 is more consistent with the characteristic that moss prefers a meta-acid environment than other treatments; the process 3 had the highest content of slime and particles and the lowest content of sand. The moisture content of the substrate varied from 61.07% to 83.65%, with the moisture content of treatment 10 being up to 83.65% and the moisture content of treatment 17 (CK) being 61.07% at the lowest.
From the above results, it can be seen that peat soil: vermiculite =2:1 and 20 percent of Knop nutrient solution is favorable for the growth of the dentritic moss and is easy to realize the large-area planting of the dentritic moss.
Example 5
After 7 days of culture of the dentures from examples 1 to 3, the growth of the dentures from examples 1 to 3 was observed and counted, and the results are shown in the following Table 4:
TABLE 4 growth results of dentures on different treatments
It can be seen that the rhodobryum dentiger grows faster than the crushed rhodobryum dentiger in the stem and leaf separation state, and is more favorable for the rapid growth of the rhodobryum dentiger.
The results show that different matrixes, treatment modes and hormones have different influences on the large-area planting of the odontopathy, the growth speed of the odontopathy can be obviously improved by the cultivation method, the photosynthesis capability of the odontopathy is improved, and the large-area planting of the odontopathy is easy to realize.
The above description is only an embodiment of the present invention, and is not intended to limit the scope of the present invention, and all equivalent structures or equivalent processes performed by the present invention or directly or indirectly applied to other related technical fields are also included in the scope of the present invention.
Claims (10)
1. A method for culturing the odontopathy is characterized in that the pretreated odontopathy is cultured on the surface of nutrient soil added with a matrix, and the matrix is a mixture of peat soil and vermiculite.
2. The culture method according to claim 1, wherein the mass ratio of the peat soil to the vermiculite is 1-3:1-3.
3. The culture method according to claim 1, wherein the rhodobryum dentium is a pulverized rhodobryum dentium or a separated stem and leaf rhodobryum dentium.
4. The cultivation method according to claim 1, wherein the pretreatment comprises 75% alcohol treatment and sterile water washing in this order, and the time for the 75% alcohol treatment is 15s to 45s.
5. The method according to claim 1, wherein a hormone is sprayed during the culturing.
6. The culture method according to claim 5, wherein the hormone is one or more of 0.01. Mu.M 6-BA, 0.1mg/L NAA, 1mg/L NAA, 0.01 mg/L2, 4-D, 0.01mg/L IAA, and 0.01mol/L IBA.
7. The culture method according to claim 1, wherein the nutrient soil is further added with a Knop nutrient solution; the Knop nutrient solution comprises 1g of calcium nitrate, 0.25g of potassium nitrate, 0.25g of monopotassium phosphate, 0.25g of magnesium sulfate, 0.003g of zinc sulfate and 1000mg of distilled water.
8. The culture method according to claim 1, wherein the culture is a membrane culture.
9. The cultivation method according to claim 1, wherein the temperature of the cultivation is 21 ℃ to 25 ℃ and the humidity of the cultivation is 50% to 55%.
10. Use of a method of culture of dentures as claimed in any one of claims 1 to 9 for the remediation of the ecological environment of a mine.
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