CN111557221A - High-efficiency propagation method for seed sources of sphagnum arenarium - Google Patents

High-efficiency propagation method for seed sources of sphagnum arenarium Download PDF

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Publication number
CN111557221A
CN111557221A CN202010299711.9A CN202010299711A CN111557221A CN 111557221 A CN111557221 A CN 111557221A CN 202010299711 A CN202010299711 A CN 202010299711A CN 111557221 A CN111557221 A CN 111557221A
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moss
propagation
crusts
matrix
inoculation
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卜崇峰
白雪强
田畅
李亚红
姚乐
韦应欣
鞠孟辰
王春
郭琦
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Northwest A&F University
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Northwest A&F University
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G22/00Cultivation of specific crops or plants not otherwise provided for
    • A01G22/30Moss
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G24/00Growth substrates; Culture media; Apparatus or methods therefor
    • A01G24/10Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G24/00Growth substrates; Culture media; Apparatus or methods therefor
    • A01G24/20Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G24/00Growth substrates; Culture media; Apparatus or methods therefor
    • A01G24/20Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
    • A01G24/22Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material containing plant material

Abstract

The invention provides a high-efficiency propagation method of a sargassum provenance, relating to the technical field of environmental engineering; taking naturally-developing moss crusts of the roughy sand as seed sources, taking disinfected sand as a substrate, adding actinomycetes and chlorella, inoculating and crushing the seed sources of the roughy moss, wherein the seed sources only need to be 0.24m2The wild moss seed source can be cultured to form 1m in 30 days under the condition of an incubator2Stable moss crusts to realize 4 times of propagation, and the coverage, the plant density and the plant height of the moss respectively reach 97.14 percent and 28.31 plants/cm22.28mm, is an effective method for efficiently cultivating the moss crusts in sand under the condition of manual control, not only provides reference for the industrial propagation practice of moss provenances in future, but also solves the problem of moss crusting field engineeringAnd a foundation is laid for the bottleneck problem of insufficient provenance recovery.

Description

High-efficiency propagation method for seed sources of sphagnum arenarium
Technical Field
The invention belongs to the technical field of environmental engineering, and particularly relates to a sargentgloryvine provenance efficient propagation method.
Background
The biological soil crust is widely distributed in arid-semiarid regions, and has important significance for local land resource maintenance and ecological environment protection especially in regions where vascular plants are difficult to survive. However, the desert biological crust is easy to be damaged, and once the desert biological crust is damaged, the desertification process can be accelerated. According to the size of the crust, the natural recovery time of a complete crust is different from several years to ten years, and the period is long.
At present, moss crust seed sources required by the restoration work of moss crust are all collected from the field, and the destruction to the local ecological environment is inevitable in the seed source collection process, and the influence of physical factors such as light, temperature, water and inoculum size or chemical factors such as nutrient solution and growth regulator on the cultivation of the moss crust is mainly researched in the past, most of the studies are concentrated in loess plateau areas and desert research areas such as Xinjiang and Ningxia, and the moss plants with remarkable wind prevention and sand fixation effects on the blackjack sand land and biological factors such as fungi and algae are reported as additives for promoting the growth and development of the moss crust. Meanwhile, no method capable of realizing large-scale artificial propagation is available at present, and a large amount of seed sources required by field engineering recovery are met.
Disclosure of Invention
In view of the above, the present invention aims to provide a rapid, effective and easily-popularized method for realizing industrial and efficient propagation of moss provenance, so as to solve the bottleneck problem of insufficient moss provenance required for field engineering biological crust recovery.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a high-efficiency indoor propagation method for sargentgloryvine provenance, which comprises the following steps:
(1) collecting moss crusts in the field, scooping the moss crusts by 1cm, drying in the shade for identifying seeds, picking out impurities, crushing moss crusts, stems and leaves, and uniformly mixing to obtain seed sources;
(2) shoveling native sandy soil 5-20 cm below the bark of the wild moss, sieving and disinfecting to obtain a matrix;
(3) filling the substrate mixed with the fungus additive into the propagation device; the fungus additive is actinomycetes;
(4) inoculating the seed source on the substrate, and spraying algae liquid on the surface of the substrate 2d after the inoculation; the algae liquid is water solution of chlorella;
(5) after the inoculation, the propagation is carried out for 30d under the conditions that the illumination intensity is 6000lx, the temperature is 20 ℃ and the air humidity is 75 percent, so as to obtain moss seed sources required by field engineering recovery.
Preferably, the drying time in shade in the step (1) is 3-5 d; and crushing the moss crusts dried in the shade into stem and leaf fragments with the length of 1-3 mm.
Preferably, the high-speed plant pulverizer used for pulverizing in the step (1) is eupatorium-2500C.
Preferably, the sieving in the step (2) is to sieve the native sandy soil with a sieve of 2mm after the native sandy soil is naturally dried in the sun.
Preferably, the sterilization in the step (2) is to place the screened sandy soil into a drying oven with the temperature of 108 ℃ for drying and sterilization for 80 min.
Preferably, the sand matrix in the step (3) is added in an amount of 2/3 according to the volume of the propagation device; the addition amount of the actinomycetes in the matrix is 1g/kg of the matrix.
Preferably, the inoculation amount of the seed source in the inoculation in the step (4) is 1020g/m2
Preferably, after the inoculation, spraying 2-3L/m2The water of (2).
Preferably, the spraying amount of the algae liquid in the step (4) is 2g/m2The concentration of chlorella in the algae liquid is 0.6 g/L.
Preferably, in the propagation expanding period in the step (5), the improved Hoagland nutrient solution is sprayed once every 6 days, and the dosage is 2.1L/m2
The invention provides a method for efficiently propagating sargentgloryvine provenance, which uses true-leaf moss (Bryum argenteum) collected from a hair-blacking vegetable sand as a provenance, adds actinomycetes and chlorella to the sand-soil matrix after disinfection, inoculates the moss provenance, and forms stable moss crusts after cultivation in a short time (30 d). The invention provides a moss provenance propagation method comprising a series of scientific and effective steps of provenance treatment, substrate disinfection, inoculation maintenance and the like, which not only provides reference for large-scale propagation of sandy moss provenance in the future, but also provides sufficient provenance and theoretical basis for field engineering recovery of moss crust.
The addition of actinomycetes and chlorella in the optimum propagation combination 'sand matrix + actinomycetes + chlorella' obviously improves the peroxidase activity and the soluble protein content of moss crust, reduces the malondialdehyde content, enhances the water retention capacity of cells, protects cell membranes and other life substances, and only needs 0.24m under the condition of an incubator (the illumination, the temperature and the air humidity are respectively set to be 6000lx, 20 ℃ and 75 percent), namely2The wild moss seed source can be cultivated to form 1m in a short time (30d)2Stable moss crusts to realize 4 times of propagation, and the coverage, the plant density and the plant height of the moss respectively reach 97.14 percent and 28.31 plants/cm2、2.28mm。
Drawings
FIG. 1 is a graph showing a comparison of the growth at the initial stage of culture (a) and at the final stage of culture (b, c) in all treatments;
FIG. 2 is a graph of moss skinning coverage, plant density and height over time for different treatments;
FIG. 3 is a graph comparing POD, soluble protein and MDA in different treatments of moss crusts 15d and 30 d.
Detailed Description
The invention provides a high-efficiency indoor propagation method for sargentgloryvine provenance, which comprises the following steps: (1) collecting moss crusts in the field, scooping the moss crusts by 1cm, drying in the shade for identifying seeds, picking out impurities, crushing moss crusts, stems and leaves, and uniformly mixing to obtain seed sources;
(2) shoveling native sandy soil 5-20 cm below the bark of the wild moss, sieving and disinfecting to obtain a matrix;
(3) filling the substrate mixed with the fungus additive into the propagation device; the fungus additive is actinomycetes;
(4) inoculating the seed source on the substrate, and spraying algae liquid on the surface of the substrate 2d after the inoculation; the algae liquid is water solution of chlorella;
(5) after the inoculation, the propagation is carried out for 30d under the conditions that the illumination intensity is 6000lx, the temperature is 20 ℃ and the air humidity is 75 percent, so as to obtain moss seed sources required by field engineering recovery.
The method comprises the steps of collecting moss crusts in the field, shoveling the moss crusts by 1cm, drying in the shade for identifying seeds, picking out impurities, crushing moss crusting stem leaves and fragments, and uniformly mixing to obtain provenance. The method preferably collects well-developed and complete moss crusts, and uses a small shovel to shovel the moss crusts by 1 cm. The collected cortex is preferably put into a gunny bag with good air permeability and transported back to a laboratory, and the gunny bag is placed in the shady place and dried for standby.
According to the method, the moss crust is preferably dried in the shade for 3-5 days and then crushed, the crushing is preferably carried out by crushing the crust into stem and leaf fragments with the lengths uniformly distributed between 1-3mm by using a high-speed plant crusher (Ezela-2500C), and the stem and leaf fragments are uniformly mixed to obtain a moss crust seed source.
The method comprises the steps of shoveling native sandy soil 5-20 cm under the bark of wild moss, sieving and disinfecting to obtain a matrix. The method preferably collects the underlying soil layer 5-20 cm deep under the heading moss crust as a sandy soil matrix (hereinafter referred to as sandy soil) during propagation of moss crust seed sources, collects the soil layer 5-20 cm deep, and can eliminate the influence of scattered propagules in the crust. The screened sandy soil is collected, preferably naturally dried in the sun, screened by a 2mm screen and placed in a shade. The disinfection of the invention is preferably to place the screened sandy soil into a drying box with the temperature of 108 ℃ for drying and disinfection for 80 min.
The substrate mixed with the fungus additive is filled in the propagation device; the fungus additive is actinomycetes. The propagation device is preferably a culture box, the specification is 17cm long, 17cm wide and 12cm high, and the bottom of the propagation device is provided with a plurality of small holes. According to the invention, before the culture box is filled with the matrix, a layer of square felt cloth which is 14cm long and wide and is sterilized by potassium permanganate diluent is preferably paved, so that the effects of matrix loss prevention and water retention can be achieved.
The present invention preferably adds 2/3 volume ratio of the matrix to the incubation box; the addition amount of the actinomycetes in the matrix is 1g/kg of the matrix. The fungus additive such as actinomycetes can improve soil nutrients.
Inoculating the seed source on the matrix, and spraying algae liquid on the surface of the matrix 2d after the inoculation; the algae solution is water solution of chlorella. In the inoculation of the invention, the seed source is preferably directly inoculated. In the inoculation of the invention, the inoculation amount of the seed source is preferably 1020g/m2. After the inoculation, the invention preferably also comprises spraying 2-3L/m2The water of (2).
2d after the inoculation, spraying a chlorella water solution on the surface of the substrate, wherein the dosage of the chlorella is 2g/m2The concentration of chlorella in the algae liquid is 0.6 g/L. It can not only secrete filament solidified soil particles, but also promote the planting development of crust through the action of carbon fixation and nitrogen fixation.
After the inoculation, the invention carries out propagation for 30d under the conditions that the illumination intensity is 6000lx, the temperature is 20 ℃ and the air humidity is 75 percent, and obtains moss provenance needed by field engineering recovery. In the propagation period, the Hoagland improved orchid nutrient solution is preferably sprayed once every 6 days, and the dosage is preferably 2.1L/m2. The invention only needs 0.24m after the cultivation for 30 days2The wild moss seed source can be cultivated to form 1m in a short time (30d)2Stable moss crusts, realizes 4 times of propagation, and has the advantages of high coverage, high plant density and high plant heightThe degrees respectively reach 97.14 percent and 28.31 strains/cm2、2.28mm。
The present invention will be described in detail with reference to examples, but the present invention is not limited to these examples.
Example 1
1. Test material
1) Lichen crusts
Moss crust samples were collected from the city of elm, west, Shaanxi province, in the Mongolian Bay county (109 ° 36 '14 "-109 ° 36' 24" E, 38 ° 32 '50 "-38 ° 32' 54" N), at an altitude of 1148-1153 m. The region belongs to a moderate-temperature-zone semiarid continental monsoon climate, the annual average temperature is 8.3 ℃, the annual average precipitation is 405mm, and the precipitation is mainly concentrated in 6-9 months. The plot is located at the southeast edge of Maousu sand, and the landform type is mainly fluctuating fixed and semi-fixed sand dunes. The dominant shrubs are Artemisia desertorum (Artemisia desertorum), salix psammophila (Salixpsammophila) and Caragana Caragana (Caragana korshinski), which are often uniformly distributed in clusters in the plot. The herbaceous plants are mainly Sasa albo-marginata (Psammochlora villosa) and Heteropappus officinalis (Heteropappus hispidus), which are distributed sporadically in the plots.
And selecting well-developed and relatively complete moss crusts during sampling, and using a small shovel to shovel the moss crusts by 1 cm. And (4) putting the collected cortex into a gunny bag with good air permeability, transporting the gunny bag back to a laboratory, and placing the gunny bag in a shady place for airing for later use. The tested moss crusts are mainly determined to be the family of the dictyotaceae, and the dominant species is the Bryum argenteum (Bryum argenteum). After drying in the shade, manually picking out macroscopic plant litter, root systems, stones, livestock manure and the like, crushing the bark layer into stem and leaf fragments with the lengths uniformly distributed between 1 mm and 3mm by using a high-speed plant crusher (Ezela-2500C), and uniformly mixing to obtain the moss bark seed source for later use.
2) Soil matrix
The soil matrix collecting place is the same as the moss crust collecting place. In order to eliminate the influence of scattered propagules in the crust layer, the crust of the overground moss and a soil layer with the thickness of 0-5 cm are removed, and then an underlying soil layer with the thickness of 5-20 cm is dug. Collecting soil, transporting to a laboratory, naturally drying in the sun, sieving by a 2mm sieve, and placing in a shade for later use. Placing the screened sandy soil, the seedling raising matrix and the organic fertilizer into a drying box with the temperature of 108 ℃ for drying and disinfection, wherein the screened sandy soil and the seedling raising matrix are dried for 80min, the organic fertilizer is dried for 40min, the organic fertilizer is taken out and turned over once midway, and the disinfected sandy soil and the organic fertilizer are placed in a shade for later use. The matrix used in the experiment comprises a formula matrix and a sand matrix, wherein the formula matrix is formed by mixing and stirring disinfected sand, a seedling culture matrix and an organic fertilizer according to the volume ratio of 1:1:0.2, and the sand matrix is the disinfected sand.
2. Experiment design method
The 8 treatments shown in table 1 were designed, and a sand matrix + seed source (treatment 9) and a formula matrix + seed source (treatment 10) were set as control groups, and each treatment was repeated 4 times.
Table 1 details of experimental treatments
3. Cultivation process and method
1) Preparation of culture Material
Firstly, placing a piece of felt cloth (the length is ×, the width is 14cm × 14cm) sterilized by potassium permanganate diluent at the bottom of a sterilized cultivating box (the length, the width and the height are respectively 17cm, 17cm and 12cm, and a plurality of small holes are arranged at the bottom), then filling a 5cm thick substrate (sand soil substrate/formula substrate) added with actinomycetes (1 g/kg substrate) or bacillus megaterium (1 g/kg substrate) according to the experimental design, leveling the surface of the substrate, uniformly broadcasting 20g seed sources (the inoculation area is 14cm × 14cm, namely 1020 g/m) in each sample2). Adding Artemisia desertorum gum (0.1 g/sample) into seed source, stirring, inoculating, spraying 50ml water to make Artemisia desertorum gum concentration reach 0.2%, and spraying equal amount of clear water for treating without Artemisia desertorum gum. Algae solution (2 g/m)2) Uniformly spreading on the sample according to the treatment requirement 2 days after the moss stem and leaf fragments are inoculated.
2) Moss crust cultivation process
The experiment was completed in an incubator. The light, temperature and air humidity of the incubator were set to 6000lx, 20 ℃ and 75%, respectively. In the cultivation period, the improved Hoagland nutrient solution is sprayed once every 6 days, and the dosage is 2.1L/m2And measuring moss crust growth indexes (coverage, density and plant height) once a week (7 days) when the new plant grows out, finishing the experiment when the crust coverage tends to be stable, and lasting for 30 days.
4. Observation index and method
1) Determination of growth indicators
Randomly measuring the heights of 20 mosses in the culture box by using an electronic vernier caliper, and taking the mean value as the plant height; uniformly selecting 5 2.5cm multiplied by 2.5cm small frames in each culture box, counting the number of moss plants in the small frames, calculating the average value, and dividing the average value by the area of one small frame to obtain the plant density in one culture box; the coverage is determined by a point needle sample frame method, the size of a square grid plate is 2.5cm multiplied by 2.5cm, the specification of the grid is 0.5cm multiplied by 0.5cm, moss is under the focus of the grid, the moss is marked as 1, the moss is not marked as 0, and the total number of points with crust under the focus is divided by the total number of points of the grid plate, so that the coverage is determined.
2) Measurement of physiological index
1. Reagent and preparation method
(1)0.2mol/L pH7.8 sodium phosphate (Na)2HPO4-NaH2PO4) Buffer solution:
liquid A (0.2mol/L NaH)2PO4Solution): 15.715g NaH was accurately weighed2PO4·2H2Dissolving O in a small 50ml beaker with a small amount of distilled water, transferring into a 500ml volumetric flask, metering to a scale with distilled water, and mixing well. Storing in a refrigerator at 4 ℃ for later use.
Liquid B (0.2mol/L Na)2HPO4Solution): 71.63g of Na are accurately weighed2HPO4·12H2Dissolving O in a small beaker of 100ml with a small amount of distilled water, transferring into a volumetric flask of 1000ml, fixing the volume to the scale with the distilled water, and fully and uniformly mixing. Storing in a refrigerator at 4 ℃ for later use.
Mixing solution A34 ml and solution B366 ml to obtain 0.2mol/L sodium phosphate buffer solution with pH 7.8. Storing in a refrigerator at 4 ℃ for later use.
(2)50mmol/L pH7.8 sodium phosphate buffer solution
Taking 250ml of 0.2mol/L sodium phosphate buffer solution with pH7.8, transferring into a 1000ml volumetric flask, fixing the volume to the scale with distilled water, and fully and uniformly mixing. Storing in a refrigerator at 4 ℃ for later use.
(3) Peroxidase (POD) reaction solution
Pouring 50ml of 100mmol/L phosphate buffer solution into a beaker, adding 28 mu L of guaiacol, placing on a magnetic stirrer for heating and stirring until the guaiacol is dissolved, adding 19 mu L of 30% H after the solution is cooled2O2And storing in a refrigerator at 4 ℃ after uniform mixing.
(4) 0.6% Thiobabital (TBA) solution
0.6g of TBA is accurately weighed, dissolved by a small amount of 1mol/L NaOH, and then the volume of 10 percent TCA solution is adjusted to 100 ml.
(5) Standard protein solution (100. mu.g/ml bovine serum albumin)
Accurately weighing 25mg of bovine serum albumin, adding distilled water to dissolve and fix the volume to 100ml, sucking 40ml, and diluting with distilled water to 100 ml.
(6) Coomassie brilliant blue reagent
100mg of Coomassie brilliant blue G-250 is accurately weighed, dissolved in 50ml of 90% ethanol, added with 85% (W/V) phosphoric acid, added with distilled water to reach the volume of 1000ml, and stored in a brown bottle. Can be stored at room temperature for one month.
2. Extraction of enzyme solution
Randomly pulling out moss stem and leaf fragments to 0.1g by using tweezers in each culture box, putting the moss stem and leaf fragments into a precooled mortar, adding 2ml of 50mmol/L sodium phosphate buffer solution (extraction medium) with pH7.8, grinding the moss stem and leaf fragments into slurry, adding 3ml of extraction medium to wash the mortar, namely 5ml of enzyme extracting solution, and directly moving the moss stem and leaf fragments into a centrifuge tube from the mortar. Placing the centrifuge tube into a 4 deg.C centrifuge, centrifuging at 8000r/min for 30min to obtain supernatant as crude enzyme extract, transferring the crude enzyme extract into 5ml centrifuge tube, and storing at 0-4 deg.C.
3. Determination of Peroxidase (POD): guaiacol method was used. Adding 100 μ L enzyme extract into 3ml POD reaction solution, and placing in ultraviolet spectrophotometerMeasuring absorbance at 470nm wavelength, reading every 1min, reading absorbance 3min before reaction, and using distilled water instead of enzyme solution as control. By Delta A470The enzyme activity is expressed in units of/g FW/min and the POD enzyme activity is calculated according to the following formula:
POD Activity (U/g FW) ═ A470×Vt)/(W×VS×t)
In the formula A470Represents the change in absorbance over the reaction time; w represents fresh weight (g); vtRepresenting the total volume of the enzyme extract; vSRepresents the volume of enzyme solution (ml) taken for the assay; t represents the reaction time (min).
4. Determination of Malondialdehyde (MDA) content: the method of spearmint barbituric acid (TBA) is used. Absorbing 1ml of enzyme extract by using a pipette gun, adding 4ml of 0.6% TBA solution, uniformly mixing, carrying out boiling water bath for 15-30 min, then rapidly cooling to room temperature, centrifuging for 10min on a 4000r/min centrifuge, and taking supernate to respectively measure the absorbance values at the wavelengths of 450nm, 523nm and 600nm on an ultraviolet spectrophotometer. The MDA content was calculated according to the following formula:
C=6.45×(A532-A600)-0.56×A450
Y=C×V/W
in the formula: c represents MDA concentration (. mu.mol/ml); y represents the MDA content (. mu. mol/g); v represents the total volume (ml) of the extract; w represents the fresh weight of the stem and leaf fragments of the moss plants.
5. Determination of soluble protein: the Coomassie brilliant blue method was used. Sucking 0.1ml of enzyme extract with a pipette, transferring into a test tube with scales, adding 0.9ml of distilled water, adding 5ml of Coomassie brilliant blue G-250 reagent, mixing thoroughly, standing for 2min, measuring absorbance at 595nm with an ultraviolet spectrophotometer, and determining protein content with a standard curve. The standard curve drawing method comprises the following steps:
adding the reagents into 6 test tubes according to the table 2, mixing uniformly, adding 5ml of Coomassie brilliant blue reagent into each tube, shaking uniformly, standing for about 5min, and performing colorimetric determination on absorbance at 595nm by using a test tube No. 0 as a blank control. And drawing a standard curve by taking the protein content as an abscissa and the absorbance as an ordinate.
TABLE 20-100 μ g Standard protein to distilled Water ratio in protein content
Soluble protein content was calculated according to the following formula:
M=C×Vt/(Vs×W)
wherein M represents the content of soluble protein in the sample (μ g/g); c represents the protein content found in the standard curve; vtRepresenting the total volume (ml) of the enzyme extract; vsRepresents the volume (ml) of enzyme extract at the time of measurement; w represents the mass (g) of the stem and leaf fragments of the moss plants.
5. Results and analysis
1) Effect of different treatments on the growth status of lichen crust
FIG. 1 shows the growth of moss crusts at the initial stage and the final stage of cultivation in all treatments, from which it can be seen that stable moss crusts can be formed after cultivation for 30 days in an incubator (light, temperature and air humidity are set to 6000lx, 20 ℃ and 75% respectively). FIG. 2 shows the changes of moss crust coverage, plant density and plant height in 10 treatments at 7 days, 15 days, 22 days and 30 days of cultivation, and it can be seen that there is moss crust development at 7 days of cultivation. For the coverage and the height, the growth rate is rapidly increased within 7-15 days, then the growth rate is gradually reduced along with the extension of the cultivation time, and the height difference among all treatments is not significant and is 1.77-2.32 mm; the strain density is opposite to the change trend, namely the strain density is gradually increased in the early period and then gradually increased. At the end of incubation, the average values of the skinning coverage and the strain density of all the treated strains of the formula substrate are respectively 22.38 percent and 4.49 strains/cm2Above, the cultivation effect of the treatment 5 (formula substrate + sand sagebrush gum + bacillus megaterium + chlorella) is excellent, and the coverage, the strain density and the mean strain height are 54.90% and 13.03 strains/cm respectively2And 2.28 mm; the average values of the crust covering degree and the plant density of the sand matrix under all treatments are 89.73 percent and 23.73 plants/cm respectively2The cultivation effect of the treatment 4 (sand matrix, actinomycetes and chlorella) is better, and the coverage, the strain density and the strain height of the treatment 4 are higherThe degree average values are respectively 97.14 percent and 28.31 strains/cm2And 2.28 mm. Comprehensively considered, all treatments can promote the growth of moss crusts, so that the cultivation effect of treating 4 (sand matrix, actinomycetes and chlorella) is optimal, and only 0.24m is needed2The wild moss seed source can be cultivated to form 1m in a short time (30d)2Stable moss crusts, and the propagation multiple reaches 4 times.
2) Mechanism of influence of exogenous additives on treatment of moss 4 crust growth
When plants are stressed by adversity, H is produced2O2Etc. active oxygen, which can cause serious damage to plant cells, and Peroxidase (POD) of plants and the like are key enzymes in an antioxidant system in plants and can effectively remove the active oxygen in the plants; the soluble protein is an important osmotic adjusting substance, has strong hydrophilicity, is obviously increased in content under stress of a certain degree, and can improve the water retention capacity of cells; malondialdehyde (MDA) is a product of membrane lipid peroxidation, is a marker substance for cell membrane destruction, and its content can indicate the degree of membrane lipid peroxidation and the strength of plant response to adverse conditions. In order to clearly determine the best reason for treating the growth of the 4 moss crusts, the relationship between 3 physiological indexes such as POD activity, soluble protein content, MDA content and the like in the moss crusts and each experimental factor in the cultivation process is analyzed.
The three physiological indexes can effectively reflect the growth condition of the moss. As can be seen from the correlation analysis (Table 3), moss POD and soluble protein have a very significant positive correlation (P <0.01) with the coverage and the strain density, and MDA has a very significant negative correlation (P <0.01) with the coverage and the strain density.
TABLE 3 correlation analysis of various indexes under different treatments
Indicates p <0.05, the correlation reached significance, indicates p <0.01, the correlation reached very significant.
FIG. 3 shows that in the incubation period, the moss skin of treatment 4 (sand matrix + actinomycetes + chlorella) is best in growth, the POD and soluble protein contents are continuously increased in the incubation period, the POD and soluble protein contents reach 108.69U/g FW and 244.36mg/g FW respectively in 30d of incubation, which are obviously higher than those of other treatments (P <0.05), and the MDA content is lower than that of other treatments and is only 34.61 mu mol/g, so that the oxygen radical scavenging capability of the moss skin is enhanced, the peroxidation degree of the cell membrane lipid of the mesophyll cells is reduced, and the aging speed is slowed down.
The anova results showed (table 4) that matrix type had a very significant effect on soluble proteins and MDA of moss crusts (P < 0.01); the influence of algae additives on POD, soluble protein and MDA of moss is very obvious (P < 0.01); the effect of fungus additives on soluble protein is very significant (P < 0.01). The matrix type and the bacteria and algae additive have obvious influence on the physiological characteristics and the growth condition of the moss crusts.
TABLE 4 analysis of variance of three physiological indexes of moss by different factors
Therefore, the treatment 4 (sandy soil matrix + actinomycetes + chlorella) with sandy soil as the matrix obviously improves the POD activity and the soluble protein content of the moss through different exogenous additives, reduces the MDA content, enhances the water retention capacity of cells, protects the living matters, further effectively increases the coverage and the strain density of the moss, and is the optimal mode for cultivating and propagating the sandy moss.
The method can quickly obtain a large amount of moss provenances and is used for restoring the wild moss crust and restoring the environment.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Claims (10)

1. An efficient indoor propagation method for sargentgloryvine provenance is characterized by comprising the following steps:
(1) collecting moss crusts in the field, scooping the moss crusts by 1cm, drying in the shade for identifying seeds, picking out impurities, crushing moss crusts, stems and leaves, and uniformly mixing to obtain seed sources;
(2) shoveling native sandy soil 5-20 cm below the bark of the wild moss, sieving and disinfecting to obtain a matrix;
(3) filling the substrate mixed with the fungus additive into the propagation device; the fungus additive is actinomycetes;
(4) inoculating the seed source on the substrate, and spraying algae liquid on the surface of the substrate 2d after the inoculation; the algae liquid is water solution of chlorella;
(5) after the inoculation, the propagation is carried out for 30d under the conditions that the illumination intensity is 6000lx, the temperature is 20 ℃ and the air humidity is 75 percent, so as to obtain moss seed sources required by field engineering recovery.
2. The method according to claim 1, wherein the drying time in shade in step (1) is 3-5 d; and crushing the moss crusts dried in the shade into stem and leaf fragments with the length of 1-3 mm.
3. The method according to claim 1, wherein the high-speed plant pulverizer used in the pulverizing of step (1) is eupatorium-2500C.
4. The method as claimed in claim 1, wherein the sieving in step (2) is carried out by drying the native sandy soil in the sun and sieving the native sandy soil by a 2mm sieve.
5. The method according to claim 1 or 4, wherein the sterilization in step (2) is carried out by placing the screened sandy soil into a drying oven at 108 ℃ for drying and sterilizing for 80 min.
6. The method according to claim 1, wherein the sand matrix in step (3) is added in an amount of 2/3 times the volume of the propagation device; the addition amount of the actinomycetes in the matrix is 1g/kg of the matrix.
7. The method of claim 1, wherein the amount of said seed source inoculated during said inoculation in step (4) is 1020g/m2
8. The method according to claim 7, characterized in that after the inoculation, the method further comprises spraying 2-3L/m2The water of (2).
9. The method as claimed in claim 1, wherein the spraying amount of the algae liquid in the step (4) is 2g/m2The concentration of chlorella in the algae liquid is 0.6 g/L.
10. The method as claimed in claim 1, wherein the improved Hoagland nutrient solution is sprayed every 6 days in the propagation period of step (5) at a rate of 2.1L/m2
CN202010299711.9A 2020-04-16 2020-04-16 High-efficiency propagation method for seed sources of sphagnum arenarium Pending CN111557221A (en)

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