CN115989732A - Method for improving drought resistance of festuca arundinacea - Google Patents
Method for improving drought resistance of festuca arundinacea Download PDFInfo
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Abstract
A method for improving drought resistance of festuca arundinacea comprises four stages of seed disinfection, breeding, seedling fixing and seedling reviving, and is characterized in that expanded arbuscular mycorrhizal fungi are added into a culture medium in the breeding step, and the arbuscular mycorrhizal fungi are one or two of young sleeve saccule mould and Mortierella morchella. In the plant seed breeding stage, arbuscular mycorrhizal fungi are added into a culture medium of seeds, so that the arbuscular mycorrhizal fungi are used for impregnating plant roots and then symbiosis, a combination is formed, the activities of CAT, SOD, POD and proline in plant plants are further enhanced, the oxidation injury resistance and permeation regulation capability of plant leaf tissues are further enhanced, and the festuca arundinacea can better cope with the damage caused by drought stress.
Description
Technical Field
The invention relates to the technical field of planting, in particular to a method for improving drought resistance of festuca arundinacea.
Background
Festuca arundinacea (Festuca arundinacea Schreb.) is also called reed festuca, is a grass family, is a perennial herb of sheep Mao Yashu, has extremely high nutritional value, is an important grass family pasture in the world, is widely applied to urban greening and sports field construction due to the characteristics of barren resistance, disease resistance, wide adaptability and the like, and shows remarkable social, ecological and environmental benefits, and festuca arundinacea is increasingly valued as turf grass for people.
However, festuca arundinacea is used as turf grass, the planting area of festuca arundinacea is continuously enlarged according to the requirements, but the festuca arundinacea is not drought-enduring like natural turf, and especially the thirst-enduring capability is poor. The root system of the lawn is shallow, which is insufficient to absorb enough moisture in the soil, and the lawn can be irrigated only manually. The lawn needs to be watered once every day in dry seasons, even in rainy seasons, the lawn needs to be watered at least twice a month, a large amount of water is needed in management, and the lawn growth is poor once the water is deficient in seasons with little rainwater or areas with relatively low water resources. Therefore, the cultivation of new varieties of drought-resistant and water-saving festuca arundinacea is urgent.
In view of this, the present application is specifically proposed.
Disclosure of Invention
The invention aims to provide a method for improving drought resistance of festuca arundinacea, which is characterized in that arbuscular mycorrhizal fungi are added into a culture medium of seeds in a festuca arundinacea seed breeding stage, so that the arbuscular mycorrhizal fungi are used for dying the festuca arundinacea root system and then symbiotic, a complex is formed, the activities of CAT, SOD, POD and proline in festuca arundinacea plants are further improved, the oxidation injury resistance and permeation regulation capacity of festuca arundinacea Mao Shepian tissues are further improved, and the festuca arundinacea can better cope with damage caused by drought stress.
Embodiments of the present invention are implemented as follows: a method for improving drought resistance of festuca arundinacea comprises a breeding stage and is characterized in that expanded arbuscular mycorrhizal fungi are added into a culture medium in the breeding stage, and the arbuscular mycorrhizal fungi are one or two of young sleeve sacculus fungi and mousse sacculus fungi.
Further, a seedling stage is provided after the breeding stage, the seedling stage comprising the step of increasing the SLs content (single angle Jin Nazhi) in the fescue plants.
Further, the step of improving the SLs content in the fescue plants in the seedling stage comprises the following steps: the gao Mao Shepian was sprayed with Shi Du feet of the golactone analog.
Further, the concentration of strigolactone analog is 0.2. Mu. Mol.L -1 The frequency of spraying is once daily for a total of 10 days.
Further, the spraying degree is that the blade is stained with dew and does not drip.
Further, a seed disinfection stage is provided before the breeding stage, the seed disinfection stage comprising: selecting festuca arundinacea seeds with full seeds and uniform sizes, soaking the festuca arundinacea seeds in 75% alcohol for 1-3min, washing the festuca arundinacea seeds with distilled water for 3-4 times, soaking the festuca arundinacea seeds in 84 disinfectant for 2-5min, and washing the festuca arundinacea seeds with distilled water for 3-4 times.
Further, the breeding stage is plug breeding, firstly, filling culture matrixes to 2/3 of each seedling tray, watering, then sowing festuca arundinacea seeds, covering a sterilized matrix of 0.5cm, watering at the bottom of each seedling tray, and then moving to an illumination culture room for culture.
Further, a seedling fixing stage is arranged before the seedling recovering stage, the seedling fixing stage comprises the step of growing the tall fescue seedlings to a two-leaf stage after 14 days of breeding, and 1/4-1/3 of seedlings are reserved in each plug tray.
Further, the method for propagating arbuscular mycorrhizal fungi mainly comprises the following steps:
(1) seed disinfection: soaking sorghum and corn seeds in 40% formaldehyde solution for 15min, and then washing with water for several times;
(2) and (3) sterilizing a substrate: selecting vermiculite and perlite matrixes according to the weight ratio of 1:1, steam sterilizing for 2 times at 95 ℃ for 1-2h each time, and spacing for 24h in the middle;
(3) culturing: filling sterilized culture medium to 2/3 of plastic basin, spreading the strain of arbuscular mycorrhizal fungi on the medium uniformly, covering with sterilized medium for 2cm, watering, sowing sorghum seeds, covering with sterilized medium for 0.5cm, transferring to illumination room, culturing for 3-4 months, and harvesting.
Further, the culture conditions in the illumination chamber mainly include illumination time: 14h white/10 h black; illumination intensity: 8000lx; temperature: 20-30 ℃; humidity: 60%.
The embodiment of the invention has the beneficial effects that:
1. according to the method for improving drought resistance of festuca arundinacea provided by the embodiment of the invention, arbuscular mycorrhizal fungi (mainly one or two of young sleeve sacculus fungi and mousse sacculus fungi) are added into a culture medium of festuca arundinacea seeds in a festuca arundinacea seed breeding stage, so that the arbuscular mycorrhizal fungi are used for dying the roots of the festuca arundinacea to form symbiotic plants, a combination is formed, the activities of CAT, SOD, POD and proline in festuca arundinacea plants are further improved, and the oxidation injury resistance and permeation regulation capacity of tissue of festuca arundinacea Mao Shepian are further improved, so that the festuca arundinacea can better cope with damage caused by drought stress.
2. According to the method for improving drought resistance of festuca arundinacea provided by the embodiment of the invention, arbuscular mycorrhizal fungi (mainly one or two of young sleeve sacculus fungi and Morse sacculus fungi) are added into a culture medium of festuca arundinacea seeds in a festuca arundinacea seed breeding stage, so that the grown festuca arundinacea can keep growth of plant height, width, leaf length and leaf root at any time under drought stress, and the festuca arundinacea plants can keep good growth situations, and can withstand the test of drought conditions on artificial lawns.
3. According to the method for improving drought resistance of festuca arundinacea provided by the embodiment of the invention, in order to further improve the activities of CAT, SOD, POD and proline in festuca arundinacea plants and also improve the content of strigolactone in the plants, shi Du foot strigolactone analogues are sprayed on leaves in the seedling-reviving stage of festuca arundinacea plants, and the activities of CAT, SOD, POD and proline in the plants are synergistically enhanced with arbuscular mycorrhizal fungi, so that the drought resistance of festuca arundinacea plants is further improved, the damage caused by drought stress can be relieved by the synergistic effect of SLs and AMF, the oxidative damage caused by drought stress can be relieved, the osmotic pressure of cell tissues can be regulated, and the stress resistance of the plants can be enhanced.
4. According to the method for improving drought resistance of festuca arundinacea provided by the embodiment of the invention, the content of strigolactone in plants can be improved by spraying Shi Du feet of strigolactone analogues to the festuca arundinacea Mao Shepian in a seedling stage, the infection effect of AMF on host plants can be enhanced by SLs in the plants, and the symbiotic relationship between the AMF and the plants is promoted.
In general, the festuca arundinacea plant cultivated by the method for improving the drought resistance of festuca arundinacea provided by the embodiment of the invention has strong drought resistance.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the embodiments will be briefly described below, it being understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and other related drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a diagram of a treated sheep Mao Biaoxing according to Experimental example 1 of the present invention;
FIG. 2 is a graph showing the SLs content of the sheep Mao Genbu after the treatment of experimental example 1;
FIG. 3 is a graph showing the effect on festuca arundinacea growth index after treatment in experimental example 1 of the present invention;
FIG. 4 is a graph showing the effect of the treatment of experimental example 1 on the biochemical index of the sheep Mao Shengli;
FIG. 5 is an AMF spore subgraph after propagation in examples 1-3 of the present invention;
FIG. 6 is a graph Mao Biaoxing of the treated sheep of Experimental example 2;
FIG. 7 is an AMF infection rate and infection chart after treatment of experimental example 2 of the present invention;
FIG. 8 is a graph showing the effect of AMF on festuca arundinacea growth after treatment according to Experimental example 2 of the present invention;
FIG. 9 is a graph showing the effect of AMF on the biochemical index of the sheep Mao Shengli after treatment in Experimental example 2;
FIG. 10 is a graph of a treated high sheep Mao Biaoxing according to Experimental example 3 of the present invention;
FIG. 11 is a graph showing the SLs content of festuca arundinacea after treatment according to Experimental example 3 of the present invention;
FIG. 12 is an AMF infection rate and infection chart after treatment of experimental example 3 of the present invention;
FIG. 13 is a graph showing the growth index of festuca arundinacea after treatment according to experimental example 3 of the present invention;
FIG. 14 is a graph showing the effect of the treatment of Experimental example 3 on the biochemical index of the sheep Mao Shengli;
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
Examples
The embodiment provides a method for improving drought resistance of festuca arundinacea, which mainly comprises three stages of seed disinfection, breeding and seedling reviving and is characterized in that expanded arbuscular mycorrhizal fungi are added into a culture medium in the breeding step, and the arbuscular mycorrhizal fungi are one or two of young sleeve sacculus fungi and mousse sacculus fungi.
The embodiment provides a method for improving drought resistance of festuca arundinacea, which mainly comprises four stages of seed disinfection, breeding, seedling setting and seedling reviving and is characterized in that arbuscular mycorrhizal fungi after propagation are added into a culture medium in the breeding step, wherein the arbuscular mycorrhizal fungi are one or two of young sleeve saccule mould and Mortierella moellendorffii.
In this example, the propagated arbuscular mycorrhizal fungi include the resulting culture medium consisting of spores, hyphae, mycorrhizal segments and culture medium.
In the scheme, the seed breeding of festuca arundinacea can be carried out in a seedling raising tray, and can be directly sowed in the land for breeding.
Further, the method also comprises the step of increasing the content of the single-horn Jin Nazhi (SLs) in the fescue plants in the seedling reviving period.
Further, the step of improving SLs in the fescue plants in the seedling stage comprises the following steps: the gao Mao Shepian was sprayed with Shi Du feet of the golactone analog.
Further, the concentration of strigolactone analog is 0.2. Mu. Mol.L -1 The frequency of spraying is once daily for a total of 10 days.
Further, the spraying degree is that the blade is stained with dew and does not drip.
Further, the seed sterilization step includes: selecting festuca arundinacea seeds with full seeds and uniform sizes, soaking the festuca arundinacea seeds in 75% alcohol for 1-3min, washing the festuca arundinacea seeds with distilled water for 3-4 times, soaking the festuca arundinacea seeds in 84 disinfectant for 2-5min, and washing the festuca arundinacea seeds with distilled water for 3-4 times.
Further, the breeding step is plug breeding, firstly, filling culture medium to 2/3 of each seedling tray, watering, then sowing festuca arundinacea seeds, covering a sterilized medium with the thickness of 0.5cm, watering at the bottom of each seedling tray, and then moving to an illumination culture room for culture.
Further, the step of setting seedlings comprises the step of growing tall fescue seedlings to a two-leaf period after 14 days of breeding, wherein 1/4-1/3 of seedlings are reserved in each plug.
Example 1
The test strain of arbuscular mycorrhizal fungi in the embodiment is a young bush sacculus mildew 1 strain (Glomus etunicatum, BEG QH01B, QH01B for short), and the strain is purchased from the institute of plant nutrition and resource of the national academy of sciences of agriculture and forestry in Beijing.
Mainly comprises the following steps:
1. seed disinfection: the seeds need to be disinfected before sowing, soaked in 40% formaldehyde solution for 15min, and then washed with water for several times.
2. And (3) sterilizing a substrate: vermiculite and perlite matrixes are selected according to the proportion of 1:1, and the sterilization of the matrixes kills the existing AMF, pathogens and weed seeds, so that no pathogenic microorganisms exist in the planting area. Steam sterilizing at 95deg.C for 1-2 hr for 2 times with interval of 24 hr, and standing for 1-2 weeks. All containers are sterilized before use, and can be soaked in 10% sodium hypochlorite (bleaching powder) solution for 30min. Cleaning and drying. Or soaking or wiping with 70-75% ethanol solution.
3. Culturing: filling sterilized culture medium to 2/3 of plastic basin, spreading the above-mentioned standby inoculant (20 g) on the medium uniformly in a thin layer, or uniformly dropping the above-mentioned standby spores (test strain) on the medium, covering sterilized medium 2cm, watering, sowing sorghum seeds 25-30 grains/basin or corn seeds 4-5 grains/basin, covering sterilized medium 0.5cm, transferring to illumination room, culturing for 3-4 months (illumination time: 14h white/10 h black; illumination intensity: 8000lx; temperature: 20-30 deg.C; humidity: 60%), and adopting Hoagland (phosphorus reduction) nutrient solution (Table 3-1-1) of Hoagland watering once every week. In harvesting, the stem and leaf of the overground part of the plant is firstly cut off, the pot is placed in a room with relatively stable temperature and humidity for drying (about 1-2 weeks), and then all cultures (including plant roots, hyphae, spores and matrixes) in the pot are harvested.
4. Harvesting: the operating table top is firstly wiped and disinfected by 70% alcohol. The tabletop was covered with 3 layers of clean paper (new newspaper or new copy paper), the cultures in the pots were poured onto the paper and chopped with a sterile short axe. The chopped cultures were wound up in two rolls, one end of the paper tube being inserted into a clean plastic bag ready for sealing. When all cultures are transferred into the human bags, the paper is gently taken out, the bags are sealed, hands do not need to contact the potted cultures at any time, the same plastic bags are sleeved outside, and special BGC labels are placed between the two plastic bags. And then placed in a refrigerator or 4 ℃ refrigerator for preservation according to the species sequence of the tags.
Example 2
The present embodiment provides a method for propagating arbuscular mycorrhizal fungi, in which the test strain of the arbuscular mycorrhizal fungi: the test strain is a young sacculus 1 strain (Glomus etunicatum, BEG QH01B, QH01B for short; moschig sacculus 1 strain (Glomus moseae, BEG KH01, KH01 for short) 2 strains purchased from the institute of plant nutrition and resource of the national academy of sciences of agriculture and forestry in Beijing city.
Mainly comprises the following steps:
1. seed disinfection: the seeds need to be disinfected before sowing, soaked in 40% formaldehyde solution for 15min, and then washed with water for several times.
2. And (3) sterilizing a substrate: vermiculite and perlite matrixes are selected according to the proportion of 1:1, and the sterilization of the matrixes kills the existing AMF, pathogens and weed seeds, so that no pathogenic microorganisms exist in the planting area. Steam sterilizing at 95deg.C for 1-2 hr for 2 times with interval of 24 hr, and standing for 1-2 weeks. All containers are sterilized before use, and can be soaked in 10% sodium hypochlorite (bleaching powder) solution for 30min. Cleaning and drying. Or soaking or wiping with 70-75% ethanol solution.
3. Culturing: filling sterilized culture medium to 2/3 of plastic basin, spreading the above-mentioned standby inoculant (20 g) on the medium uniformly in a thin layer, or uniformly dropping the above-mentioned standby spores on the medium, covering sterilized medium 2cm, watering, sowing sorghum seed 25-30 grains/basin or corn seed 4-5 grains/basin, covering sterilized medium 0.5cm, transferring to illumination room to culture for 3-4 months (illumination time: 14h white/10 h black; illumination intensity: 8000lx; temperature: 20-30 deg.C; humidity: 60%), adopting Hoagland (phosphorus reduction) nutrient solution (Table 3-1-1), and pouring the nutrient solution once every week, 30-100 ml/basin. In harvesting, the stem and leaf of the overground part of the plant is firstly cut off, the pot is placed in a room with relatively stable temperature and humidity for drying (about 1-2 weeks), and then all cultures (including plant roots, hyphae, spores and matrixes) in the pot are harvested.
4. Harvesting: the operating table top is firstly wiped and disinfected by 70% alcohol. The tabletop was covered with 3 layers of clean paper (new newspaper or new copy paper), the cultures in the pots were poured onto the paper and chopped with a sterile short axe. The chopped cultures were wound up in two rolls, one end of the paper tube being inserted into a clean plastic bag ready for sealing. When all cultures are transferred into the human bags, the paper is gently taken out, the bags are sealed, hands do not need to contact the potted cultures at any time, the same plastic bags are sleeved outside, and special BGC labels are placed between the two plastic bags. And then placed in a refrigerator or 4 ℃ refrigerator for preservation according to the species sequence of the tags.
Example 3
The present embodiment provides a method for propagating arbuscular mycorrhizal fungi, wherein the strain of the arbuscular mycorrhizal fungi is a Moschiomyces 1 strain (Glomus moseae, BEG KH01, abbreviated as KH 01), and the strain is purchased from the institute of plant nutrition and resources of the national academy of sciences of agriculture and forestry in Beijing.
Mainly comprises the following steps:
1. seed disinfection: the seeds need to be disinfected before sowing, soaked in 40% formaldehyde solution for 15min, and then washed with water for several times.
2. And (3) sterilizing a substrate: vermiculite and perlite matrixes are selected according to the proportion of 1:1, and the sterilization of the matrixes kills the existing AMF, pathogens and weed seeds, so that no pathogenic microorganisms exist in the planting area. Steam sterilizing at 95deg.C for 1-2 hr for 2 times with interval of 24 hr, and standing for 1-2 weeks. All containers are sterilized before use, and can be soaked in 10% sodium hypochlorite (bleaching powder) solution for 30min. Cleaning and drying. Or soaking or wiping with 70-75% ethanol solution.
3. Culturing: filling sterilized culture medium to 2/3 of plastic basin, spreading the above-mentioned standby inoculant (20 g) on the medium uniformly in a thin layer, or uniformly dropping the above-mentioned standby spores on the medium, covering sterilized medium 2cm, watering, sowing sorghum seed 25-30 grains/basin or corn seed 4-5 grains/basin, covering sterilized medium 0.5cm, transferring to illumination room to culture for 3-4 months (illumination time: 14h white/10 h black; illumination intensity: 8000lx; temperature: 20-30 deg.C; humidity: 60%), adopting Hoagland (phosphorus reduction) nutrient solution (Table 3-1-1), and pouring the nutrient solution once every week, 30-100 ml/basin. In harvesting, the stem and leaf of the overground part of the plant is firstly cut off, the pot is placed in a room with relatively stable temperature and humidity for drying (about 1-2 weeks), and then all cultures (including plant roots, hyphae, spores and matrixes) in the pot are harvested.
4. Harvesting: the operating table top is firstly wiped and disinfected by 70% alcohol. The tabletop was covered with 3 layers of clean paper (new newspaper or new copy paper), the cultures in the pots were poured onto the paper and chopped with a sterile short axe. The chopped cultures were wound up in two rolls, one end of the paper tube being inserted into a clean plastic bag ready for sealing. When all cultures are transferred into the human bags, the paper is gently taken out, the bags are sealed, hands do not need to contact the potted cultures at any time, the same plastic bags are sleeved outside, and special BGC labels are placed between the two plastic bags. And then placed in a refrigerator or 4 ℃ refrigerator for preservation according to the species sequence of the tags.
Example 4
The embodiment provides a method for improving drought resistance of festuca arundinacea, which adopts festuca arundinacea seeds of Hunting dog No. five as a material and mainly comprises the following steps:
1. seed disinfection: selecting festuca arundinacea seeds with full seeds and uniform sizes. Soaking in 75% alcohol for 1min, washing with distilled water for 3-4 times, soaking in 84 disinfectant for 2min, and washing with ultrapure water for 3-4 times.
2. Breeding, namely, adopting seedling raising trays (4X 8 grids, total 32 grids, upper caliber: 60mm, bottom 28mm and depth 53 mm), firstly filling test matrixes (nutrient soil: vermiculite=1:1) to 2/3 of each seedling raising tray, uniformly spreading AM fungi subjected to propagation in the embodiment 1 on a thin layer of the matrixes, covering about 20g of the matrixes, sterilizing the matrixes by 2cm, watering, sowing 6-8 grains/box of festuca arundinacea seeds, covering 0.5cm of the sterilized matrixes, pouring tap water at the bottom of each seedling raising tray, and then moving to an illumination culture room for culture.
3. And (3) seedling establishment: after 14 days, tall fescue seedlings grew to the two-leaf stage, with two seedlings per box remaining.
4. And (3) seedling reviving: watering and carrying out seedling reviving treatment.
Example 5
The embodiment provides a method for improving drought resistance of festuca arundinacea, which adopts festuca arundinacea seeds of Hunting dog No. five as a material and mainly comprises the following steps of
1. Seed disinfection: selecting festuca arundinacea seeds with full seeds and uniform sizes. Soaking in 75% alcohol for 1min, washing with distilled water for 3-4 times, soaking in 84 disinfectant for 2min, and washing with ultrapure water for 3-4 times.
2. Breeding, namely, adopting seedling raising trays (4X 8 grids, total 32 grids, upper caliber: 60mm, bottom 28mm and depth 53 mm), firstly filling test matrixes (nutrient soil: vermiculite=1:1) to 2/3 positions of each seedling raising tray, uniformly spreading AM fungi subjected to propagation in the embodiment 2 on a thin layer of the matrixes, covering about 20g of the matrixes, sterilizing the matrixes by 2cm, watering, sowing 6-8 grains/box of festuca arundinacea seeds, covering 0.5cm of the sterilized matrixes, pouring tap water at the bottom of each seedling raising tray, and then moving to an illumination culture room for culture.
3. And (3) seedling establishment: after 14 days, tall fescue seedlings grew to the two-leaf stage, with two seedlings per box remaining.
4. And (3) seedling reviving: watering and carrying out seedling reviving treatment.
Example 6
The embodiment provides a method for improving drought resistance of festuca arundinacea, which adopts festuca arundinacea seeds of Hunting dog No. five as a material and mainly comprises the following steps of
1. Seed disinfection: selecting festuca arundinacea seeds with full seeds and uniform sizes. Soaking in 75% alcohol for 1min, washing with distilled water for 3-4 times, soaking in 84 disinfectant for 2min, and washing with ultrapure water for 3-4 times.
2. Breeding, namely, adopting seedling raising trays (4X 8 grids, total 32 grids, upper caliber: 60mm, bottom 28mm and depth 53 mm), firstly filling test matrixes (nutrient soil: vermiculite=1:1) to 2/3 positions of each seedling raising tray, uniformly spreading AM fungi subjected to propagation in the embodiment 3 on a thin layer of the matrixes, covering about 20g of the matrixes, sterilizing the matrixes by 2cm, watering, sowing 6-8 grains/box of festuca arundinacea seeds, covering 0.5cm of the sterilized matrixes, pouring tap water at the bottom of each seedling raising tray, and then moving to an illumination culture room for culture.
3. And (3) seedling establishment: after 14 days, tall fescue seedlings grew to the two-leaf stage, with two seedlings per box remaining.
4. And (3) seedling reviving: watering and carrying out seedling reviving treatment.
Example 7
The embodiment provides a method for improving drought resistance of festuca arundinacea, which adopts festuca arundinacea seeds of Hunting dog No. five as a material and mainly comprises the following steps of
1. Seed disinfection: selecting festuca arundinacea seeds with full seeds and uniform sizes. Soaking in 75% alcohol for 1min, washing with distilled water for 3-4 times, soaking in 84 disinfectant for 2min, and washing with ultrapure water for 3-4 times.
2. Breeding, namely, adopting seedling raising trays (4X 8 grids, total 32 grids, upper caliber: 60mm, bottom 28mm and depth 53 mm), firstly filling test matrixes (nutrient soil: vermiculite=1:1) to 2/3 of each seedling raising tray, uniformly spreading AM fungi subjected to propagation in the embodiment 1 on a thin layer of the matrixes, covering about 20g of the matrixes, sterilizing the matrixes by 2cm, watering, sowing 6-8 grains/box of festuca arundinacea seeds, covering 0.5cm of the sterilized matrixes, pouring tap water at the bottom of each seedling raising tray, and then moving to an illumination culture room for culture.
3. And (3) seedling establishment: after 14 days, tall fescue seedlings grew to the two-leaf stage, with two seedlings per box remaining.
4. And (3) seedling reviving: watering, and spraying 0.2 mu mol L-1 strigolactone analogue (prepared by dissolving strigolactone analogue (GR 24) in acetone) on leaves of the seedling; the foliar spraying is continuously carried out every day, the spraying degree is that the foliar is stained with dew and does not drip, the soil moisture is kept sufficient, and the spraying is continued for 10 days.
Comparative example 1
The embodiment provides a method for improving drought resistance of festuca arundinacea, which adopts festuca arundinacea seeds of Hunting dog No. five as a material and mainly comprises the following steps:
1. seed disinfection: selecting festuca arundinacea seeds with full seeds and uniform sizes. Soaking in 75% alcohol for 1min, washing with distilled water for 3-4 times, soaking in 84 disinfectant for 2min, and washing with ultrapure water for 3-4 times.
2. Breeding, namely adopting seedling raising trays (4X 8 grids, total 32 grids, upper caliber: 60mm, bottom 28mm and depth 53 mm), firstly loading test matrixes (nutrient soil: vermiculite=1:1) to 2/3 of each seedling raising tray, then covering 2cm of sterilization matrixes, watering, sowing 6-8 grains/box of festuca arundinacea seeds, covering 0.5cm of sterilization matrixes, pouring tap water at the bottom of each seedling raising tray, and then moving to an illumination culture room for culture.
3. And (3) seedling establishment: after 14 days, tall fescue seedlings grew to the two-leaf stage, with two seedlings per box remaining.
4. And (3) seedling reviving: watering and carrying out seedling reviving treatment.
Comparative example 2
The embodiment provides a method for improving drought resistance of festuca arundinacea, which adopts festuca arundinacea seeds of Hunting dog No. five as a material and mainly comprises the following steps of
1. Seed disinfection: selecting festuca arundinacea seeds with full seeds and uniform sizes. Soaking in 75% alcohol for 1min, washing with distilled water for 3-4 times, soaking in 84 disinfectant for 2min, and washing with ultrapure water for 3-4 times.
2. Breeding, namely, adopting seedling raising trays (4X 8 grids, total 32 grids, upper caliber: 60mm, bottom 28mm and depth 53 mm), firstly filling test matrixes (nutrient soil: vermiculite=1:1) to 2/3 of each seedling raising tray, uniformly spreading AM fungi subjected to propagation in the embodiment 1 on a thin layer of the matrixes, covering about 20g of the matrixes, sterilizing the matrixes by 2cm, watering, sowing 6-8 grains/box of festuca arundinacea seeds, covering 0.5cm of the sterilized matrixes, pouring tap water at the bottom of each seedling raising tray, and then moving to an illumination culture room for culture.
3. And (3) seedling establishment: after 14 days, tall fescue seedlings grew to the two-leaf stage, with two seedlings per box remaining.
4. And (3) seedling reviving: watering, and spraying pure water on leaves of seedlings; the foliar spraying is continuously carried out every day, the spraying degree is that the foliar is stained with dew and does not drip, the soil moisture is kept sufficient, and the spraying is continued for 10 days.
Comparative example 3
The embodiment provides a method for improving drought resistance of festuca arundinacea, which adopts festuca arundinacea seeds of Hunting dog No. five as a material and mainly comprises the following steps of
1. Seed disinfection: selecting festuca arundinacea seeds with full seeds and uniform sizes. Soaking in 75% alcohol for 1min, washing with distilled water for 3-4 times, soaking in 84 disinfectant for 2min, and washing with ultrapure water for 3-4 times.
2. Breeding, namely, adopting seedling raising trays (4X 8 grids, total 32 grids, upper caliber: 60mm, bottom 28mm and depth 53 mm), firstly filling test matrixes (nutrient soil: vermiculite=1:1) to 2/3 of each seedling raising tray, uniformly spreading AM fungi subjected to propagation in the embodiment 1 on a thin layer of the matrixes, covering about 20g of the matrixes, sterilizing the matrixes by 2cm, watering, sowing 6-8 grains/box of festuca arundinacea seeds, covering 0.5cm of the sterilized matrixes, pouring tap water at the bottom of each seedling raising tray, and then moving to an illumination culture room for culture.
3. And (3) seedling establishment: after 14 days, tall fescue seedlings grew to the two-leaf stage, with two seedlings per box remaining.
4. And (3) seedling reviving: watering and spraying Shi Du foot-lactone on the leaves of seedlingsPreparation 5. Mu. Mol.L -1 (a strigolactone inhibitor (TIS 108) is dissolved in acetone; the foliar spraying is continuously carried out every day, the spraying degree is that the foliar is stained with dew and does not drip, the soil moisture is kept sufficient, and the spraying is continued for 10 days.
Experimental example 1 Effect of strigolactone on drought resistance of festuca arundinacea
1.1 materials and reagents
1.1.1 plant Material
Festuca arundinacea with "Hunting dog five-size" is used as plant material and planted in greenhouse (with illumination intensity of 700 mu mol.m) -2 s -1 The temperature is 20-25 ℃, the illumination time is 14h, the darkness time is10 h, and the humidity is 60%)
1.1.2 reagents
Weighing 100mg of strigolactone inhibitor (TIS 108), adding 10mL of acetone, stirring until all the inhibitor is dissolved, and preparing into a solution with concentration of 10mg.mL -1 Is preserved in dark at 4 ℃ for standby. Weighing 10mg strigolactone analog (GR 24), adding 10mL acetone, stirring until all the mixture is dissolved, and preparing into 1 mg/mL -1 Is preserved in dark at 4 ℃ for standby; SOD, CAT and POD kits from Shanghai's pullup biotechnology limited were used.
1.2 design of experiments
Selecting festuca arundinacea seeds with full seeds and uniform sizes. Soaking in 75% alcohol for 1min, washing with distilled water for 3-4 times, soaking in 84 disinfectant for 2min, washing with ultrapure water for 3-4 times, and germinating in a constant temperature incubator at 30deg.C. After the seeds naturally germinate, selecting seedlings with good growth and consistent growth vigor, and raising the seedlings by using seedling raising plug trays (with the upper caliber of 60mm, the bottom of 28mm and the depth of 53 mm), wherein when the substrates in the seedling raising plug trays are cultured to a two-leaf period according to the proportion of nutrient soil to vermiculite of 1:1, a blank control group (CK) and 5 mu mol.L-1 TIS108 (optimal concentration screened by a preliminary pre-test) are arranged for test treatment, and three biological repetitions are treated for each treatment to obtain 360 strains. The foliar spray is continuously carried out at 15:00 a day, the spraying degree is that the foliar is full of dew and does not drip, the soil moisture is kept sufficient, and the treatment is continued for 7d. According to the characteristic change of the leaf, the continuous drought treatment is carried out for 15d in a natural state, and then the water is re-soaked for 5d, so that the physiological and biochemical indexes of the plant are measured.
1.3 test methods
1.3.1 festuca arundinacea plant phenotype
The phenotype of the sheep Mao Waiguan before and after drought treatment was photographed by a single-lens reflex camera (model: canon EOS 6D) and the photographs were simply processed by PS software.
1.3.2 measurement of festuca arundinacea growth index
The gao Mao Zhugao, root length, leaf length (fifth leaf), leaf width (fifth leaf) were measured using a ruler (precision 0.1 cm) and vernier caliper (precision 0.01 cm), and the data were recorded.
1.3.3 measurement of the amount of Gaoguang Mao Shengwu
And taking out the treated festuca arundinacea plants from the culture dish, cleaning soil attached to the roots, sucking water by filter paper, weighing on a balance, and recording fresh weight. And (3) rapidly placing the processed fresh weight into an oven for de-enzyming for 20min at 110 ℃, adjusting the oven to 60 ℃, baking for 24h until the weight is constant, weighing on a balance, and recording the dry weight.
1.3.4 measurement of Biochemical index of festuca arundinacea
Selecting the functional leaves of the sheep Mao Di to 7, wiping off surface stains, quick-freezing with liquid nitrogen, and storing at the temperature of-80 ℃ for later use.
1.3.5 festuca arundinacea SLs content determination
1.3.6 statistical analysis
Statistical analysis of the data was performed using Excel 2010 and plotted, correlation analysis and one-way analysis of variance (ANOVA) were performed using SPSS24.0 software, and multiple comparisons were performed by Duncan's method at α=0.05, with results expressed as mean ± standard error.
1.4 experimental results and analysis
1.4.1 Effects of SLs on festuca arundinacea phenotypic characteristics
As can be seen from fig. 1: spraying 5 mu mol.L under drought condition -1 The drought tolerance of festuca arundinacea after the inhibitor TIS108 is obviously reduced, and the characteristics of obvious leaf wilting and curling sagging, scorching of leaf tips and leaf bases, shortened root length and the like are shown, probably because the leaf is extremely lack of water and fails to timely under continuous drought stressAnd watering. And the Control (CK) and the high sheep Mao Shepian grow well, and the drought phenotype is obviously weakened.
1.4.2 Effect of SLs on festuca arundinacea SLs content
As can be seen from FIG. 2, after detecting the content of sheep Mao Genbu SLs (5-DS) by using a liquid chromatography-mass spectrometry method and comparing the content of sheep Mao Penshi SLs inhibitor TIS108 under drought conditions with a Control (CK), the content of sheep Mao Genbu SLs is reduced by 75.5 percent, and the difference is obvious. The reduction of root SLs content indicates that SLs plays an important role in drought resistance of festuca arundinacea.
1.4.3 Effect of SLs on festuca arundinacea growth index
As can be seen from fig. 3: after TIS108 is an inhibitor of Mao Penshi SLs of the goats under drought stress, the external morphological structure characteristics of the plants are obviously changed, and the goats Mao Shekuan, root length, fresh weight and dry weight indexes are obviously different from those of a Control (CK), so that the leaf length and the plant height are not obviously different (P < 0.05). Leaf width is reduced by 30.4%, root length is reduced by 19.7% compared with CK, fresh weight is 0.7 times of that of the control, and dry weight is reduced by 48.7% compared with CK. Spraying the inhibitor causes the morphological indexes of the festuca arundinacea to change obviously, which shows that SLs has important influence on drought resistance of festuca arundinacea.
1.4.4 Effect of SLs on the Biochemical index of Gaoguang Mao Shengli
In drought stress, plant antioxidant enzymes (CAT, POD, SOD) act synergistically with each other, and can remove active oxygen in plants or repair macromolecules damaged by oxidation, and the activity intensity of the plant antioxidant enzymes is closely related to the plant antioxidant capacity. As can be seen from fig. 4: after the SLs inhibitor TIS108 is sprayed, the difference between the gaofu Mao Shepian SOD, POD, CAT under drought stress and the Control (CK) is obvious, and the difference is respectively reduced by 11.3%, 22.9% and 24.8%. The spraying of the inhibitor greatly reduces the activities of CAT, POD and SOD enzymes of the sheep Mao Shepian, so that oxidative damage of drought stress to the sheep Mao Shepian cannot be relieved.
Table 1 high sheep Mao Cunhuo rate
Table 2-5-1 Fescue survival rate
Note that: the difference between different lower case letters representing the same index is significant (P < 0.05)
Note:Different lowercase letters represent significant differences between the same indicators(P<0.05)
Experimental example 2 Effect of arbuscular mycorrhizal fungi on drought resistance of festuca arundinacea
1.1 materials and reagents
1.1.1 Experimental materials
Experimental reagent: the Shanghai Chamaecypic Biotechnology Co., ltd. SOD, CAT, POD and proline kit were selected.
1.2 test procedure
Experiments were carried out with (1) comparative experiment 1, (2) example 4, (3) example 6, (4) example 5, and four treatment groups, seedlings at the final stage were used. The tall fescue seedlings grow to a two-leaf period after 14 days of seedling setting, two seedlings are reserved per box, and three biological replicates are treated each time, and 768 plants are remained. And (3) watering normally in the early stage, carrying out seedling reviving treatment, growing for 15 days, and measuring physiological and biochemical indexes of the leaves after continuous drought treatment for 14 days in a natural state according to the characteristic change of the leaves.
1.3 test methods
1.3.1 AMF spore detection
AM fungal spores were isolated by wet screen decantation-sucrose centrifugation.
1.3.2 AMF infection detection
The infection rate is required to be dyed. The infection rate measurement comprises the following steps: fixation, transparency, staining, flaking, observation and estimation
1.3.3 measurement of festuca arundinacea growth index
The method for detecting the growth index (plant height, leaf length, leaf width, root length, dry weight and fresh weight) of festuca arundinacea is provided.
1.3.4 determination of the Biochemical index of Gaoguang Mao Shengli
Detection 1.3.5 statistical analysis of physiological and Biochemical index (SOD, POD, CAT) of festuca arundinacea
Statistical analysis of the data was performed using Excel 2010 and plotted, correlation analysis and one-way analysis of variance (ANOVA) were performed using SPSS24.0 software, and multiple comparisons were performed by Duncan's method at α=0.05, with results expressed as mean ± standard error. The influence of different types of AMFs on the drought resistance of festuca arundinacea is comprehensively evaluated by adopting a membership function analysis method, and the method is referred to as Wang Mengke (2021).
The index formula of positive correlation: yi= (Xi-Ximin)/(Ximax-Ximin)
Index formula of negative correlation: yi=1- (Xi-Ximin)/(Ximax-Ximin)
Wherein Yi is the membership function value of the ith index under a certain process, xi is the index under a certain process, and Ximax and Ximin are the maximum value and the minimum value of the ith index in all the processes respectively.
1.4 results and analysis
1.4.1 AMF propagation
As can be seen from fig. 5: after 4 treatment group strains KH01 and QH01B are propagated by sorghum and corn, spores and hyphae in a culture medium are clearly visible, a foundation is laid for the subsequent AMF drought resistance experiment, and the subsequent experiment can be ensured to be carried out smoothly.
1.4.2 Effects of AMF on the phenotypic characteristics of festuca arundinacea
As can be seen from fig. 6: after the festuca arundinacea is inoculated with KH01 strain (example 6) and QH01B strain (example 4) under drought stress and the two strains are mixed and inoculated (example 5), the growth vigor of the festuca arundinacea Mao Shepian is good, and the drought phenotype is obviously weakened. The drought tolerance of comparative example 1 is remarkably reduced, and the characteristics of obvious wilting and sagging of the leaves, scorch of leaf tips and leaf bases, short plant height, short root length and the like are shown, probably because the leaves are extremely lack of water and can not be watered in time under continuous drought stress. Wherein the drought resistance effect of the inoculated QH01B strain plants (the plants of the example 4) is optimal.
1.4.3 Effect of AMF on the infection of Gaoge Mao Jungen
As can be seen from fig. 7: after the festuca arundinacea is cultured by AMF, AMF mycorrhizas in root systems are clearly visible, and the infection rate is 33.37% in the example 6 group, 52.6% in the example 4 group and 35.55% in the example 5 group respectively. The infection rate of the group of the example 4 is highest, the amplification is largest, and the difference is obvious. The result shows that QH01B bacteria have the best effect on the mycorrhiza infection of festuca arundinacea under drought stress.
1.4.4 Influence of AMF on festuca arundinacea growth index
As can be seen from fig. 8: after the festuca arundinacea is inoculated with different types of AMFs under drought stress, the morphological and structural changes of the festuca arundinacea are inconsistent. The sheep Mao Zhugao, leaf width, leaf length, root length, fresh weight and dry weight were significantly different from the comparative examples (P < 0.05); wherein, in example 6, 17.3%, 37.6%, 35.1%, 17.3%, 72.3%, 74.1% are respectively added; example 4 increased 39.9%, 88.0%, 61.6%, 31.2%, 173.3%, 153.8%, respectively; example 5 increased by 10.0%, 41.7%, 32.8%, 15.1%, 20.7%, 30.4%, respectively; it can be seen that inoculating QH01B species was significantly better than other treatment groups.
1.4.5 Influence of AMF on the Biochemical index of Gaoguang Mao Shengli
The drought resistance of the plant is positively correlated with the drought resistance of the plant by the drought resistance enzyme system and the content of osmotic adjusting substances of the plant under drought stress. As can be seen from fig. 9: after AMF inoculation under drought conditions, the content of high sheep Mao Shepian CAT, SOD, POD, proline was significantly different (P < 0.05) compared to Control (CK). Wherein CAT activity: example 6, example 4, example 5 increased by 60.3%, 154.9%, 130.6%, respectively; SOD activity: example 6, example 4, example 5 increased by 53.2%, 173.1%, 104.1%, respectively; POD activity: example 6, example 4, example 5 increased by 49.2%, 63.9%, 50.9%, respectively; proline content: example 6, example 4, example 5 increased by 29.0%, 64.8%, 40.0%, respectively; the inoculated QH01B strain is superior to other treatment groups, and the tissue of the tall fescue Mao Shepian has stronger oxidation injury resistance and permeation regulation capability, so that the tall fescue can better cope with the damage caused by drought stress.
Experimental example 3 Effect of strigolactone and arbuscular mycorrhizal fungi interaction on drought resistance of festuca arundinacea
1.1 materials and reagents
1.1.1 Experimental materials
Plant material: festuca arundinacea with "Hunting dog five-size" is used as plant material and planted in greenhouse (with illumination intensity of 700 mu mol.m) -2 s -1 The temperature is 20-25 ℃, the illumination time is 14h, the darkness time is10 h, and the humidity is 60 percent).
1.2 design of experiments
Experiment is carried out together with (1) comparative example 2; (2) example 7; (3) in comparative example 3, three treatment groups, each of which was selected from seedlings of fescue grown to two leaf stages after 14 days of fixed seedling, each treatment was repeated biologically, 64 seedlings were repeated biologically, and 576 seedlings were sprayed continuously on leaf surfaces 15:00 a day to such an extent that leaves were stained with dew and did not drip, and after 10 days of continuous spraying, immediately after 15 days of continuous drought treatment according to the change of leaf characteristics, the physiological and biochemical indexes were measured after 5 days of rehydration.
1.3 test methods
1.3.1 festuca arundinacea plant phenotype
The method of section 2.3.1 is referred to by gao sheep Mao Biaoxing.
1.3.2 detection of infection by Gaoge Mao Jungen
The method of reference section 3.3.2 is infested with gao sheep Mao Jungen.
1.3.3 measurement of festuca arundinacea growth index
Festuca arundinacea growth index (root length, plant height, leaf width, leaf length, dry weight, fresh weight) refers to methods in section 1.3.2-2.3.3.
1.3.4 determination of the Biochemical index of Gaoguang Mao Shengli
The biochemical index (SOD, POD, CAT, proline) of the gaofang Mao Shengli is referred to the method of section 3.3.4.
1.3.5 festuca arundinacea SLs content determination
Methods of the above-ground and below-ground fescue fractions for SLs content are referred to in section 2.3.5.
1.3.6 statistical analysis
Statistical analysis of the data was performed using Excel 2010 and plotted, correlation analysis and one-way analysis of variance (ANOVA) were performed using SPSS24.0 software, and multiple comparisons were performed by Duncan's method at α=0.05, with results expressed as mean ± standard error.
1.4 results and analysis
1.4.1 characteristics of Gaog Mao Biaoxing
Spraying 5 mu mol.L on festuca arundinacea inoculated with AMF under drought stress -1 TIS108 and 0.2. Mu. Mol.L -1 GR24 is shown in fig. 10: the group 7 of the high sheep Mao Shepian grew better and the drought phenotype was significantly reduced. The comparative example 3 group of festuca arundinacea has obviously reduced drought tolerance, and shows the characteristics of obvious wilting and curling sagging of the leaves, scorch of the leaf tips and leaf bases, shortened root length and the like. Therefore, the drought resistance effect of the plants can be enhanced for the AMF inoculated high sheep Mao Penshi GR24 under the drought condition.
1.4.2 festuca arundinacea SLs content
Spraying 5 mu mol.L on festuca arundinacea inoculated with AMF under drought stress -1 TIS108 and 0.2. Mu. Mol.L -1 GR24, comparison of the SLs (5-DS) content of the overground and underground portions of festuca arundinacea, as can be seen from FIG. 11: the SLs content of the overground and underground parts of the group 7 is obviously higher than that of the group 2, and is increased by 99.86 percent and 84.19 percent; the SLs content of the upper and lower parts of the treatment group of comparative example 3 is significantly lower than that of the treatment group of comparative example 2, and is reduced by 56.12% and 48.97%. From the experimental results, the SLs content of the aerial part was changed more than that of the underground part.
1.4.3 high sheep Mao Jungen infestations
As can be seen from fig. 12: spraying 5 mu mol.L on festuca arundinacea inoculated with AMF under drought stress -1 TIS108 and 0.2. Mu. Mol.L -1 The AM mycorrhiza in the root system of the fescue GR24 and the hypha are clearly visible. The infection rate of the three treatment groups is 45.0%, 53.5% and 66.0% respectively. Among them, the example 7 group has the highest infection rate to the fescue root system, the comparative example 3 group has the lowest infection rate to the root system, and both the two are different from the comparative example 2 group obviously. The SLs can strengthen the infection effect of AMF on host plants and promote the AMF to form symbiotic relationship with the plants.
1.4.4 festuca arundinacea growth index
As can be seen from fig. 13: spraying 5 mu mol.L on festuca arundinacea inoculated with AMF under drought stress -1 TIS108 and 0.2. Mu. Mol.L -1 GR24, plant height, leaf width, leaf length, root length, fresh weight, dry weight of festuca arundinacea were significantly different from the control (P<0.05 A) is provided; wherein example 7 increased 32.35%, 5.89%, 28.73%, 17.94%, 25.33%, 28.65%, respectively. Comparative example 3 ComponentsReduced by 16.49%, 10.35%, 25.08%, 23.7%, 24.24%, 35.91%, respectively, and the difference between both treatment groups was significant from comparative example 2. It shows that under drought stress, SLs and AMF act synergistically to relieve damage caused by drought stress.
1.4.5 Biochemical index of Gaoguang Mao Shengli
The changes in antioxidant enzyme and proline content were different for plants under different treatments, as can be seen from fig. 14: spraying 5 mu mol.L on festuca arundinacea inoculated with AMF under drought stress -1 TIS108 and 0.2. Mu. Mol.L -1 The SOD, CAT, POD enzyme activity and proline content of GR24, festuca arundinacea were significantly different from the control (P<0.05 A) is provided; wherein the comparative example 3 group was reduced by 36.75%, 36.31%, 78.88%, 31.17%, respectively; example 7 increased 75.29%, 38.60%, 77.51%, 21.11%, respectively. Under drought stress, SLs and AMF cooperate to relieve oxidative damage caused by drought stress, regulate osmotic pressure of cell tissues and enhance stress resistance of plants.
Table 2 high sheep Mao Cunhuo rate
Table 4-5-1 Fescue survival rate
Note that: the difference between different lower case letters representing the same index is significant (P < 0.05)
Note:Different lowercase letters represent significant differences between the same indicators(P<0.05)
The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A method for improving drought resistance of festuca arundinacea comprises a breeding stage and is characterized in that expanded arbuscular mycorrhizal fungi are added into a culture medium in the breeding stage, and the arbuscular mycorrhizal fungi are one or two of young sleeve sacculus fungi and mousse sacculus fungi.
2. The method for improving drought resistance of festuca arundinacea according to claim 1, wherein a seedling stage is further provided after said breeding stage, said seedling stage comprising the step of improving the content of strigolactone in festuca arundinacea plants.
3. The method for improving drought resistance of festuca arundinacea according to claim 2, wherein the step of improving the content of strigolactone in festuca arundinacea plants in the seedling stage comprises the following steps: the gao Mao Shepian was sprayed with Shi Du feet of the golactone analog.
4. The method for improving drought resistance of festuca arundinacea according to claim 3, wherein said strigolactone analogue is present at a concentration of 0.2 μmol.L -1 The frequency of spraying is once daily for a total of 10 days.
5. The method for improving drought resistance of festuca arundinacea according to claim 4, wherein said spraying is performed to such an extent that leaves are stained with dew and do not drip.
6. The method for improving drought resistance of festuca arundinacea according to any one of claims 1-5, characterized in that a seed disinfection stage is further provided before said breeding stage, said seed disinfection stage comprising: selecting festuca arundinacea seeds with full seeds and uniform sizes, soaking the festuca arundinacea seeds in 75% alcohol for 1-3min, washing the festuca arundinacea seeds with distilled water for 3-4 times, soaking the festuca arundinacea seeds in 84 disinfectant for 2-5min, and washing the festuca arundinacea seeds with distilled water for 3-4 times.
7. The method for improving drought resistance of festuca arundinacea according to any one of claims 1-5, wherein the breeding stage is plug breeding, firstly filling culture medium to 2/3 of each seedling tray, watering, then sowing festuca arundinacea seeds, covering with 0.5cm sterilization medium, watering at the bottom of the seedling tray, and then moving to an illumination culture room for culture.
8. The method for improving drought resistance of festuca arundinacea according to any one of claims 1-5, wherein a final stage is arranged before the seedling-recovering stage, wherein the final stage comprises the period from the growth of festuca arundinacea seedlings to two leaves after 14 days of breeding, and each tray retains 1/4-1/3 of seedlings.
9. The method for improving drought resistance of festuca arundinacea according to claim 1, wherein the method for propagating arbuscular mycorrhizal fungi mainly comprises the following steps:
(1) seed disinfection: soaking sorghum and corn seeds in 40% formaldehyde solution for 15min, and then washing with water for several times;
(2) and (3) sterilizing a substrate: selecting vermiculite and perlite matrixes according to the weight ratio of 1:1, steam sterilizing for 2 times at 95 ℃ for 1-2h each time, and spacing for 24h in the middle;
(3) culturing: filling sterilized culture medium to 2/3 of plastic basin, spreading the strain of arbuscular mycorrhizal fungi on the medium uniformly for a thin layer, covering with sterilized medium for 2cm, watering, sowing sorghum seeds, covering with sterilized medium for 0.5cm, and transferring to illumination room for culturing for 3-4 months;
(4) harvesting.
10. The method for improving drought resistance of festuca arundinacea according to claim 9, wherein the culture conditions in said illumination chamber mainly comprise illumination time: 14h white/10 h black; illumination intensity: 8000lx; temperature: 20-30 ℃; humidity: 60%.
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