Rapid cultivation method and application of drought-tolerant racomitum canescens biological crust

Abstract

The invention discloses a rapid cultivation method and application of drought-tolerant racomitum canescens biological crust. The method comprises the following steps: recovering the activity of moss crust, and collecting to obtain moss stem and leaf bodies; disinfecting the moss stem and leaf bodies, inoculating the disinfected moss stem and leaf bodies into a BG11 solid culture medium for tissue culture to obtain moss protonemata, inoculating the moss protonemata into a culture medium added with 0.5 mg / L IAA and 0.1 mg / L 6-BA for aerobic culture to obtain proliferated moss protonemata; the method comprises the following steps: inoculating moss protonema to the surface of desert sandy soil, covering a breathable film, and carrying out biological crust induction culture to obtain complete moss crust. The crust can reduce the average particle size of soil by 17 microns, increase the total organic carbon content by 31%, maintain the relative moisture content of 25.8% or above under drought stress and the photosynthetic efficiency recovery rate of 98% after rehydration, has the characteristics of environmental protection and high efficiency, and significantly improves the performance of desert soil.

Description

Technical Field

[0001] The invention belongs to the technical field of ecological restoration, and specifically relates to a rapid cultivation method and application of a drought-tolerant Ceratocystis biocrust. Background Art

[0002] Biological soil crust is a special aggregate formed on the soil surface by cryptophytes such as cyanobacteria, lichens, and mosses, as well as bacteria, fungi, and other organisms in the soil. Biological soil crust is widely present in desert areas, accounting for about 30%-70% of the surface coverage. It not only plays a key role in the ecosystem, but also provides important support for soil improvement and vegetation restoration. Moss crust represents the highest stage of biological crust succession and has more advantages than other crusts in improving soil stability and inhibiting water evaporation. In addition, moss is a typical hydrophyte with strong drought tolerance and can adapt to the hot and dry extreme environment of desert areas. However, under natural conditions, the development of moss is very slow, and it often takes several years or even decades to develop into a complete moss crust. Therefore, studying the artificial proliferation method of moss, so as to achieve the rapid cultivation of moss crust, is of great significance for the ecological restoration of desert areas.

[0003] Bryophytes have unique reproductive characteristics, and they can be sexually propagated by sporophytes, and can also be asexually propagated by the protonema grown from the nutrient tissues such as broken stems and leaves. Protonema is a special stage that bryophytes are different from other higher plants. It grows from the spores, stems and leaves of mosses, and a protonema can be differentiated into countless gametophytes, thereby providing conditions for the rapid propagation of mosses. At present, the cultivation of moss crust mostly adopts the mode of "field sampling-air-dried broken skin-inoculation culture", for example, invention patent CN113287478A discloses a rapid cultivation method of moss crust, which is mainly collected by field moss crust, and moss stem and leaf fragments are obtained after rubbing and sieving, and then the moss crust is obtained by sowing and cultivating. For another example, invention patent CN104380959 discloses a rapid cultivation method of moss crust in the Loess Plateau, after it collects the well-developed moss crust in the Loess Plateau and air-dries it, it is crushed by a plant crusher, and then inoculated and cultivated to obtain moss crust. Although the above method is simple and efficient, it relies on native moss crusts collected in the wild as the seed source, which not only requires a large amount of biomass, but also damages the surface of the original habitat, so it is not suitable for large-scale desertification soil restoration. In addition, although the invention patent CN102257954B proposes a method for cultivating and preserving the protonema of Erythromyces serrulate, it does not explore its application in inducing biological soil crusts, and the cultivation cycle of this method is relatively long, which has certain difficulties in practical application.

[0004] Therefore, it is necessary to develop a cultivation method for moss crusts with fast growth rate, good repair effect and strong tolerance. Summary of the invention

[0005] The purpose of the present invention is to provide a rapid cultivation method and application of drought-tolerant Ceratium biocrust, which can improve soil structure, reduce particle size by 17 μm, increase total organic carbon by 31% and nutrient levels, enhance soil drought tolerance, maintain soil relative moisture content ≥25.8% under drought stress, and photosynthetic efficiency recovery rate ≥98% after rehydration.

[0006] In order to achieve the above object, the present invention adopts the following technical solution:

[0007] In a first aspect of the present invention, a method for rapid cultivation of drought-tolerant Ceratium biocrust is provided, characterized in that the method comprises:

[0008] After the activity of the complete moss crust is restored, the moss stem and leaf bodies are obtained by collection;

[0009] The moss stems and leaves are sterilized and then inoculated into a BG11 solid culture medium for tissue culture to obtain moss protonema;

[0010] The moss protonema is inoculated into a 1 / 2MS liquid culture medium supplemented with 0.5 mg / L IAA+0.1 mg / L 6-BA and cultured with aeration to obtain proliferated moss protonema;

[0011] The moss protonema is inoculated on the surface of desert sand and covered with a breathable film for biological crust induction culture to obtain a complete moss crust.

[0012] In the technical solution,

[0013] The complete moss of the embodiment of the present invention is Ceratocystis, which was collected from the Kubuqi Desert Desert Algae Fixation Experimental Area (44°21′N; 109°51′E). A well-developed moss crust patch with Ceratocystis as the founding species was selected, and some robust Ceratocystis plants were selected.

[0014] Preferably, the activity recovery method is to soak the moss crust for 24 hours in a completely rehydrated manner, and then cut the stems and leaves with clean scissors.

[0015] Preferably, the disinfection method is: using 20% ​​by volume H 2 O 2 Sterilize for 30 seconds, then wash three times with sterile water. The medium used is BG11 medium, the amount of agar added is 1.5% W / W, and the pH is adjusted to 5.5-6.5. The tissue culture conditions are set as follows: temperature 20-25℃, light intensity 35μmol·m-2·s-1, light-dark ratio 12:12, and culture period is 20 days.

[0016] Preferably, in the aerated culture, the culture conditions are a temperature of 25°C, a light intensity of 50 μmol·m-2·s-1, a light-dark ratio of 12:12, a ventilation volume of 0.5-1 L / min, and the gas introduced is a mixture of 3% CO and air, that is, aeration culture in a mixed gas environment containing 3% CO.

[0017] Preferably, in the induction culture of the biological crust, the culture conditions are a temperature of 25°C, a light intensity of 80-100 μmol·m-2·s-1, and a light-dark ratio of 12:12. Preferably, in the induction culture of the biological crust, a transparent plastic film with a thickness of about 0.03 mm is covered on the seedling pot, and small holes are pierced on the surface of the plastic film at a density of 150-200 per square meter to ensure the ventilation in the seedling pot, and water is added to each seedling pot every other day.

[0018] The inoculation amount of the moss protonema is 55-65 g / m 2 .

[0019] The pore density of the breathable film is 150-200 pores / m 2 .

[0020] Preferably, the soil is Kubuqi Desert soil collected in the wild, and the main component of the soil is SiO 2 , the average soil particle size is about 200μm.

[0021] In a second aspect of the present invention, the use of moss crusts cultivated by the method in desert ecological restoration is provided. The effects include:

[0022] (1) Soil improvement: the average particle size was reduced from 190.76 μm to 173.92 μm, and the proportion of fine particles increased by 7.4%;

[0023] (2) Nutrient improvement: total organic carbon increased by 31% (5.13 g / kg vs 4.05 g / kg), and urease activity increased by 73%;

[0024] (3) Enhanced drought resistance: After 96 hours of drought, the RWC is still maintained at 2%, and the Fv / Fm recovery rate reaches 98%.

[0025] The drought condition is continuous drought for 14 days under the condition of air humidity of 25%-35%.

[0026] Compared with the prior art, the present invention has the following beneficial effects:

[0027] The rapid cultivation method of drought-tolerant Ceratium biocrust provided by the present invention has the following advantages:

[0028] (1) Double-factor synergistic culture: The combination of 0.5mg / L IAA + 0.1mg / L 6-BA makes the chlorophyll a content reach 3.44mg / L (compared with the control +75%). By adding plant growth regulators IAA and 6-BA to the 1 / 2MS liquid culture medium, the effect of synergistically promoting the proliferation of Ceratium hornularis was achieved, and the formation of a stable moss crust was induced in a short period of time. This method significantly increased the proliferation rate of Ceratium hornularis, greatly reduced the dependence on natural moss resources, and had significant environmental advantages.

[0029] (2) Rapid crusting technology: Stable crusting is formed in 60 days ( Figure 2 ), the soil particle size decreased from 190.76 μm to 173.92 μm (Table 8). The above-mentioned horn moss quickly (60 days) formed a crust on the desert soil surface, reduced the soil particle size, and improved the nutrient level. In addition, the moss crust cultivated by the present invention has good water retention and drought resistance, indicating that it can better adapt to the extreme environment of drought in desert areas and has great application potential in ecological restoration in arid areas.

[0030] (3) Significant ecological benefits: total organic carbon increased by 31% (5.13 vs 4.05 g / kg); sucrase activity increased by 134% (3.41 vs 1.31 mg / g); drought tolerance: RWC maintained at 25.8% (only 6% for bare sand). BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 This is the tissue culture process of Ceratocystis spp.

[0032] Figure 2 To induce the growth condition of moss crust;

[0033] Figure 3 is the change of soil relative water content (RWC) under drought stress with drought time;

[0034] Figure 4 The changes in chlorophyll a fluorescence efficiency of crust during drought stress and rewatering. DETAILED DESCRIPTION

[0035] The present invention will be further described below in conjunction with specific examples and accompanying drawings, but the following examples are only used to illustrate the present invention and should not be considered to limit the scope of the present invention. In the following examples, if specific conditions are not specified, they are carried out under normal conditions. The methods used are conventional methods known in the art unless otherwise specified, and the consumables and reagents used are commercially available unless otherwise specified. Unless otherwise specified, the professional and scientific terms used herein have the same meaning as those familiar to those skilled in the art. In addition, any method or material similar or equivalent to the recorded content may also be applied to the present invention.

[0036] Example 1 Disinfection method of horn moss and selection of culture medium

[0037] (1) Collection of stems and leaves of Ceratocystis

[0038] Use clean scissors to cut the tender stems and leaves of the rehydrated moss plant, wash them several times with sterile water until there are no other impurities on the surface of the moss, and soak the obtained moss stems and leaves in sterile water for later use.

[0039] (2) Disinfection methods of Trichoderma and selection of culture medium

[0040] In a sterile operating hood, use 20% by volume of H 2 O 2 , 5% sodium hypochlorite and 75% ethanol for 30 seconds, then transfer the moss stems and leaves to sterile water, and wash the moss stems and leaves with sterile water three times until there is no disinfectant residue on the plant surface. Carefully pick up the moss stems and leaves with sterile tweezers and inoculate them into sterilized BG11, Hoagland, 1 / 2MS and Knop solid plates, inoculate 6-9 moss stems and leaves on each plate, and repeat three times for each treatment, and then seal the culture dish with sealing film.

[0041] The sealed solid plate was transferred to the greenhouse for cultivation. The cultivation conditions were: temperature 20-25°C, light intensity 35 μmol·m-2·s-1, light-dark ratio 12:12, cultivation period 20 days, and the growth of moss was observed. The growth of moss in each treatment group is shown in the following table:

[0042] Table 1 Growth of mosses in different disinfection methods and solid culture medium

[0043]

[0044]

[0045] From the above table, we can see that using 20% ​​H 2 O 2 The moss sterilized and inoculated in BG11 solid medium grew well, and abundant moss protonema grew within 20 days. The protonema from BG11 solid medium was collected and transferred to liquid medium for aeration culture.

[0046] Similarly, BG11, Hoagland, 1 / 2MS and Knop liquid culture media were selected for culture as shown in the following table.

[0047] Table 2 Chlorophyll a content and contamination status of different liquid culture media after 20 days of culture

[0048]

[0049] As shown in Table 2, the growth condition of 1 / 2MS liquid medium is better. After 20 days of aeration culture at 25℃, light intensity of 50μmol·m-2·s-1, and light-dark ratio of 12:12, a large number of moss protonema were obtained. The specific growth conditions are shown in the attached Figure 1 shown.

[0050] Example 3 Effect of plant growth regulators on the expansion of Ceratocystis spp.

[0051] (1) Effects of different concentrations of a single plant growth regulator on the proliferation of Ceratocystis spp.

[0052] Three plant growth regulators, vitamin B1, indoleacetic acid IAA and cytokinin 6-benzylaminoadenine (6-BA), were selected to explore their effects on promoting the proliferation of moss protonema in 1 / 2MS liquid culture medium.

[0053] The preparation method and concentration gradient of the mother solution of plant growth regulator are as follows: ① Take 0.1g vitamin B1 and 0.1g IAA and dissolve them in 100mL sterile water to obtain 1.0g / L vitamin B1 and 1.0g / L IAA mother solutions, and add them to 500mL 1 / 2MS sterilized liquid culture medium at concentrations of 0, 0.1, 0.2, 0.3, 0.4, and 0.5mg / L, respectively. ② Take 0.01g 6-BA and dissolve it in 100mL sterile water to obtain 100mg / L 6-BA mother solution, and add it to 500mL 1 / 2MS sterilized liquid culture medium at concentrations of 0.02, 0.04, 0.06, 0.08, and 0.10mg / L. The mother solutions of plant growth additives were filtered through a 0.22μm filter membrane before being added to the culture medium.

[0054] 2 mL of the moss protonema culture solution in Example 2 was inoculated into 1 / 2MS liquid culture medium with different concentrations of plant growth regulators added, and cultured for 20 days under the same culture conditions, after which the chlorophyll a content of the moss was measured. After 20 days of aeration culture at a temperature of 25°C, a light intensity of 130 μmol·m-2·s-1, and a light-dark ratio of 12:12, the chlorophyll a content of each treatment group is shown in Tables 3 to 6.

[0055] Table 3 Chlorophyll a content (mg / L) in different vitamin B1 concentration treatments on the 20th day

[0056]

[0057] Table 4 Chlorophyll a content (mg / L) in different IAA concentration treatments on the 20th day

[0058]

[0059] Table 5 Chlorophyll a content (mg / L) in different 6-BA concentration treatments on the 20th day

[0060]

[0061] It can be seen from Table 3 that vitamin B1 has no obvious effect on the proliferation content of Ceratium, and the chlorophyll a content of Ceratium shows irregular changes with increasing concentration. Therefore, vitamin B1 is not selected as an additive for subsequent Ceratium culture.

[0062] As shown in Table 4, with the increase of IAA concentration, the chlorophyll a content of Ceratium erythrorhizium first increased and then decreased, and when the IAA concentration was 0.5 mg / L, the chlorophyll a content reached a maximum value of 2.43 mg / L. Therefore, an IAA addition concentration of 0.5 mg / L was selected as the subsequent experimental condition.

[0063] As shown in Table 5, with the increase of 6-BA concentration, the chlorophyll a content of Ceratium erythrorhizon showed a trend of first increasing and then remaining basically unchanged. When the 6-BA addition amount reached 0.10 mg / L and above, the chlorophyll a content of Ceratium erythrorhizon was basically maintained at about 2.50 mg / L. Based on the principle of saving materials, the 6-BA addition amount of 0.10 mg / L was selected as the subsequent experimental condition.

[0064] (2) Effects of adding compound plant growth regulators on the proliferation of Trichoderma

[0065] Based on the experimental results of single plant growth regulators, we further explored whether the addition of compound plant growth regulators had a synergistic effect on the growth of C. hornbillium. The experimental treatment groups were set as follows: ① blank control: 1 / 2MS medium without plant growth regulators; ② 1 / 2MS medium with only 0.5mg / L IAA; ③ 1 / 2MS medium with only 0.10mg / L 6-BA; ④ 1 / 2MS medium with both 0.5mg / L IAA and 0.10mg / L 6-BA. C. hornbillium was cultured under the above four treatment conditions for 20 days according to the above inoculation method and culture conditions, and the chlorophyll a content in the culture solution of C. hornbillium was measured at 0, 5, 15 and 20 days.

[0066] Table 6 Chlorophyll a content of each treatment group during cultivation (mg / L)

[0067]

[0068] As shown in Table 6, the addition of 0.5 mg / L IAA and 0.10 mg / L 6-BA promoted the proliferation rate of Ceratium erythrorhizium, and on the 5th day, the chlorophyll a content reached 1.56 and 1.73 times that of the blank control group, respectively. On the 20th day of cultivation, the chlorophyll a content reached 1.75 and 1.79 times that of the blank control group.

[0069] It is worth noting that when IAA and 6-BA were added in mixture, they showed a synergistic effect on the proliferation of Ceratium sphaerocephalum. The chlorophyll a content on the 30th day reached 1.37 and 1.34 times that of the treatment group with 0.5 mg / L IAA and the treatment group with 0.10 mg / L 6-BA, respectively, indicating that the combined addition has a stronger promoting effect on the proliferation of Ceratium sphaerocephalum.

[0070] Therefore, according to this ratio, the culture solution of Ceratium erythrorhizium in treatment group ④ was transferred into a 10L aeration bottle to continue to expand the culture, and the culture conditions were the same as described above.

[0071] Example 4 Application of Induced Ceratium Biocrust in Improving Soil Nutrient Levels and Reducing Soil Particle Size

[0072] (1) Inoculation of moss

[0073] Remove the culture medium from the moss protonema suspension obtained by the above culture, wash it with sterile water three times, and then dry it in a cool and ventilated place until there is no water left on the surface. Spread the sand evenly in a seedling pot with a length of 33.5 cm and a width of 26.5 cm, and moisten it with a small amount of sterile water to a fresh weight of 55-65g / m 2 The moss was evenly inoculated onto the soil surface with an inoculation amount of . At the same time, bare sand without any biological material was used as a blank control. The above seedling pots were placed in a greenhouse for cultivation. A layer of transparent plastic film with a thickness of about 0.03 mm was covered on the seedling pots, and small holes were pierced on the surface of the plastic film at a density of 150-200 per square meter to ensure the air permeability in the seedling pots. The temperature during the cultivation period was 20-25°C, the light intensity was 80-100 μmol·m-2·s-1, the light-dark ratio was 12:12, and 400 mL of water was added to each seedling pot every other day. After 60 days of cultivation, a relatively complete moss crust was obtained. The growth status of the moss is shown in the attached figure. Figure 1 .

[0074] (2) Changes in chlorophyll a content and soil physical and chemical properties

[0075] On the 0th and 60th day after inoculation, soil samples were collected and mixed using a five-point sampling method with a ring knife. The upper crust samples were used to determine the chlorophyll a content, and the lower soil samples were used to determine the soil physical and chemical properties. The chlorophyll a content was determined by ethanol extraction; the total organic carbon content in the soil was determined by potassium dichromate oxidation-spectrophotometry; the ammonium nitrogen content in the soil was determined by potassium chloride extraction-spectrophotometry; the available phosphorus content in the soil was determined by sodium bicarbonate extraction-molybdenum antimony anti-spectrophotometry; the sucrase content in the soil was determined by 3,5-dinitrosalicylic acid colorimetry; the urease activity in the soil was determined by sodium phenol-sodium hypochlorite colorimetry; and the phosphatase activity in the soil was determined by disodium phenyl phosphate method. The results are shown in Table 7.

[0076] As shown in Table 7, the biomass of the moss treatment group increased significantly after 60 days of cultivation, and the chlorophyll a content reached 8.38 μg / cm 2 , while the chlorophyll a content of the control group did not change significantly compared with day 0. After 60 days of inoculation, the total organic carbon and ammonium nitrogen contents of the soil in the moss treatment group increased, and the activities of sucrase and urease were higher than those in the control group. This shows that the moss crusts were successfully propagated within 60 days, and the inoculation of mosses effectively improved the nutrient level of the soil and increased the activity of soil enzymes.

[0077] Table 7 Chlorophyll a content and soil physicochemical properties of the control group and induced moss crust

[0078]

[0079] (3) Changes in soil particle size after incubation

[0080] After the incubation, 5 g of air-dried crust samples were taken after removing plant residues. The particle size of the crust soil was measured using a laser particle size analyzer (Microtrac MRB, USA), and the mean particle size diameter (MWD) of aggregates and the percentage of soil particles of each size were calculated. The changes in soil particle size after 60 days of incubation are shown in Table 8.

[0081] Table 8 Changes in soil particle size after 20 days of cultivation

[0082]

[0083] As shown in Table 8, compared with the bare sand treatment group, the formation of moss crust reduced the average soil particle size by about 17 μm, the content of ultrafine sand and fine sand increased by 1.08% and 6.31%, respectively, and the percentage of medium sand decreased by 7.39%. The increase of fine particles in the soil is conducive to improving soil water holding capacity and promoting the formation of soil aggregates, thereby enhancing stability and reducing wind and water erosion. In addition, fine particles have higher cation exchange capacity, can better retain nutrients and improve soil fertility. Therefore, it can be seen that inducing the formation of moss crust can effectively improve soil properties.

[0084] Example 5 Application of inducing moss crust in improving soil drought tolerance

[0085] 1. Drought stress and rehydration experiment

[0086] The moss crust and bare sand cultured for 60 days in Example 2 above were soaked in water for 24 hours to reach the saturated soil moisture content. The samples were then placed in an incubator at a temperature of 25° C. and an air humidity of 15% and dried continuously until they were completely dehydrated. The dehydrated samples were dried for another 7 days and then completely rehydrated. During the dehydration and rehydration treatments, the soil moisture content and chlorophyll fluorescence efficiency were measured at regular intervals.

[0087] Soil moisture content was determined by the weight difference between wet soil weight and dry soil weight, and soil relative moisture content (RWC) was the ratio of real-time moisture content to saturated moisture content. The photosynthetic activity of the crust was determined by darkening the crust for 15 min and then measuring its photosynthetic activity with a portable plant efficiency meter (Handy PEA) with a light intensity set to 3000 μmol·m -2 s -1 , the fluorescence signal recording time is 1 s.

[0088] 2. The changes in soil moisture content and chlorophyll fluorescence efficiency are shown in Tables 9 and 10.

[0089] Table 9 Changes in soil relative water content (RWC) during drought stress (%)

[0090]

[0091] From Table 9 and Appendix Figure 2 It can be seen that under drought conditions, the water loss rate of moss crusts is significantly lower than that of bare sand. After the onset of drought stress, the soil moisture content of the bare sand group dropped rapidly. After 24 hours of stress, the RWC of the bare sand group dropped to 6.0%, while the RWC of the moss crust remained at 25.8%. The logarithmic fitting of the change of RWC over time showed that the time for the RWC of the bare sand group to drop to 1% was 36.6 hours, while the time for the RWC of the moss crust to drop to 1% was 67.1 hours, indicating that the inoculation of moss significantly improved the water holding capacity of the soil.

[0092] Table 10 Changes of Fv / Fm during drought stress and rehydration

[0093]

[0094] Fv / Fm is the maximum photochemical quantum yield of the photosystem II of photosynthetic organisms, which can be used to reflect the chlorophyll fluorescence efficiency of photosynthetic organisms. Figure 3It can be seen that within the first 24 hours of drought stress, the Fv / Fm in both treatment groups showed a trend of first increasing and then continuously decreasing. After 24 hours of drought, the Fv / Fm value in bare sand dropped to 0.01, while the Fv / Fm of moss crust approached 0 after 96 hours of drought. After 24 hours of rehydration, the Fv / Fm value of moss crust quickly recovered to the level before drought, while the Fv / Fm of bare sand only recovered to 56.5% before drought, indicating that moss crust can adapt well to drought environment, not only maintaining high photosynthetic activity after long-term dryness, but also quickly recovering photosynthetic performance after rehydration.

[0095] Example 6: Comparison between the method of the present invention and the conventional method

[0096] In order to more intuitively compare the method of the present invention with the traditional method, the test results are statistically shown as follows:

[0097] Table 11

[0098]

[0099] In summary, the present invention has the following advantages:

[0100] (1) The cultivation cycle is greatly shortened

[0101] Traditional method: relying on the inoculation of natural moss crusts, which are dried and crushed, and need 2-3 years of natural development to form a stable crust;

[0102] The method of the present invention: through rapid proliferation of protomes in liquid culture, combined with cultivation in a controlled environment, a complete biological crust can be formed in only 60 days;

[0103] Improvement rate: >98%, production efficiency increased by two orders of magnitude, meeting the needs of large-scale planting in desert ecological restoration projects.

[0104] (2) Biomass and photosynthetic activity increased significantly

[0105] Traditional method: The chlorophyll a content is only 1.48 mg / L, reflecting the slow growth of the strain and low photosynthetic efficiency;

[0106] The method of the present invention: by adding plant growth regulators (0.5 mg / L IAA + 0.1 mg / L 6-BA) to synergistically stimulate the proliferation of protonema, the chlorophyll a content reaches 3.44 mg / L;

[0107] Improvement rate: +75%, indicating that the metabolic activity of the strain is enhanced, providing stronger biological drive for soil carbon fixation and nutrient cycling.

[0108] (3) Outstanding effect of soil structure improvement

[0109] Traditional method: The soil particle size is not regulated, and the average particle size of desert sand (190.76 μm) is difficult to form stable aggregates;

[0110] The method of the present invention: after the moss crust is formed, the roots secrete polysaccharide mucus to wrap the sand particles, and the average particle size of the soil is reduced to 173.92 μm;

[0111] Improvement: -17% (increase in the proportion of fine particles), effectively improving soil water holding capacity (porosity increased by 22%) and reducing the risk of wind erosion.

[0112] (4) Excellent drought tolerance

[0113] Traditional method: The stability of crust under extreme drought has not been verified, and the RWC of bare sand approaches 0 after 72 hours of drought;

[0114] The method of the present invention: the moss crust maintains 25.8% RWC after 72 hours of drought by regulating stomatal opening and closing and osmotic regulating substance accumulation;

[0115] Improvement rate: +600% (compared to bare sand), the photosynthetic efficiency (Fv / Fm) recovery rate reached 98% after 30 minutes of rehydration, demonstrating excellent drought resistance and recovery ability.

[0116] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are illustrative and are not to be construed as limitations on the present invention. A person skilled in the art may make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention, which shall all be included in the scope of protection of the present invention.

Claims

1. A method for rapid cultivation of drought-tolerant Ceratium biocrusts, characterized in that: The method comprises: After the activity of the complete moss crust is restored, the moss stem and leaf bodies are obtained by collection; The moss stems and leaves are sterilized and then inoculated into a BG11 solid culture medium for tissue culture to obtain moss protonema; The moss protonema is inoculated into a 1 / 2MS liquid culture medium supplemented with 0.5 mg / L IAA+0.1 mg / L 6-BA and cultured with aeration to obtain proliferated moss protonema; The moss protonema is inoculated on the surface of desert sand and covered with a breathable film for biological crust induction culture to obtain a complete moss crust.

2. The rapid cultivation method of drought-tolerant Ceratium biocrust according to claim 1, characterized in that: The activity recovery method comprises: completely rehydrating the moss crust by soaking it and then cutting the stems and leaves.

3. The rapid cultivation method of drought-tolerant Ceratium biocrust according to claim 1, characterized in that: The moss stems and leaves are disinfected by using 20% ​​by volume H2O2 for 30 seconds, and then washed with sterile water for 3 times.

4. The rapid cultivation method of drought-tolerant Ceratium biocrust according to claim 1, characterized in that: In the tissue culture, the culture conditions are a temperature of 25° C., a light intensity of 35 μmol·m-2·s-1, and a light-dark ratio of 12:

12.

5. The rapid cultivation method of drought-tolerant Ceratium biocrust according to claim 1, characterized in that: In the aeration culture, the culture conditions are a temperature of 25° C., a light intensity of 50 μmol·m-2·s-1, a light-dark ratio of 12:12, and a ventilation volume of 0.5-1 L / min.

6. The method for rapid cultivation of drought-tolerant Ceratium biocrust according to claim 1, characterized in that: The inoculation amount of the moss protonema is 55-65 g / m 2 .

7. The rapid cultivation method of drought-tolerant Ceratium biocrust according to claim 1, characterized in that: The pore density of the breathable film is 150-200 pores / m 2 .

8. The method for rapid cultivation of drought-tolerant Ceratium biocrust according to claim 1, characterized in that: In the biological crust induction culture, the culture conditions are a temperature of 20-25° C., a light intensity of 80-100 μmol·m-2·s-1, a light-dark ratio of 12:12, and water is added to each seedling pot every other day.

9. Use of the moss crust cultivated by the method according to any one of claims 1 to 8 in desert ecological restoration.

10. The use according to claim 9, characterized in that: In the application, the moss crust improves the soil, increases nutrients, and enhances drought resistance.

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