CN113207590A - Bletilla direct seeding and seedling raising method utilizing symbiotic bacteria - Google Patents
Bletilla direct seeding and seedling raising method utilizing symbiotic bacteria Download PDFInfo
- Publication number
- CN113207590A CN113207590A CN202110471049.5A CN202110471049A CN113207590A CN 113207590 A CN113207590 A CN 113207590A CN 202110471049 A CN202110471049 A CN 202110471049A CN 113207590 A CN113207590 A CN 113207590A
- Authority
- CN
- China
- Prior art keywords
- bletilla striata
- bletilla
- symbiotic bacteria
- seeds
- direct seeding
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/25—Root crops, e.g. potatoes, yams, beet or wasabi
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
- A01C1/06—Coating or dressing seed
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C21/00—Methods of fertilising, sowing or planting
- A01C21/005—Following a specific plan, e.g. pattern
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
- A01G24/12—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material containing soil minerals
- A01G24/15—Calcined rock, e.g. perlite, vermiculite or clay aggregates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
- A01G24/28—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material containing peat, moss or sphagnum
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
Abstract
The invention discloses a bletilla direct seeding and seedling raising method by utilizing symbiotic bacteria, which specifically comprises the following steps: s1 obtaining bletilla striata symbiotic bacteria: culturing bletilla striata explants to obtain bletilla striata symbiotic bacteria strains for later use; s2 direct seeding and seedling raising of bletilla striata: seed mixing is carried out on the pretreated bletilla striata seeds and the bletilla striata symbiotic bacteria in the step S1, and then the seeds are sowed in a seedbed; s3 sowing later-stage management: after the sowed seeds sprout out, fertilizing and hardening the seedlings, and then transplanting. The bletilla striata symbiotic bacteria and bletilla striata seeds are mixed uniformly and then are subjected to direct seeding and seedling hardening treatment, the bletilla striata symbiotic bacteria are added in the bletilla striata seed germination process, the direct seeding germination rate of the bletilla striata seeds is up to 86%, a Huabao foliar fertilizer is sprayed in the later management process, and the seedling growth speed is high.
Description
Technical Field
The invention belongs to the technical field of seedling culture of traditional Chinese medicinal materials, and particularly relates to a bletilla direct seeding seedling culture method by utilizing symbiotic bacteria.
Background
Bletilla striata (thunb. ex a. murray) rchb. f., also known as Bletilla striata, a perennial herb of the genus Bletilla of the family orchidaceae. Bletilla striata is a rare Chinese herbal medicine in China and has the effects of astringing to stop bleeding, reducing swelling and promoting tissue regeneration. Chemical analysis shows that the tuber of bletilla contains various organic compounds such as bibenzyl, polysaccharides, dihydrophenanthrene, polyphenols, flavonoids and the like; modern pharmacological studies show that the organic compounds have various biological activities of resisting cancer, resisting oxidation, treating cardiovascular diseases and the like. The bletilla striata gum rich in bletilla striata can be used as a natural suspension, a thickening agent, a lubricating agent, an emulsifying agent, a humectant and the like to be widely applied to the industrial fields of cosmetics, daily chemicals, food and the like besides the medicinal value of the bletilla striata, and has huge application prospect.
Bletilla striata has strict requirements on growth environment conditions, wild bletilla striata is mostly distributed in hills, low mountain stream valley sides and hidden grass clumps or under-forest wetlands with higher altitude, and is naturally distributed in a plurality of provinces of Gansu, Sichuan, Guizhou and the like in China. In the early years, bletilla striata medicinal materials are mainly mined by wild resources, the wild bletilla striata is excessively mined, the wild resources are reduced rapidly, bletilla striata seeds are fine and have no endosperm, so that the germination rate is low under natural conditions, the growth is slow, the traditional propagation mode is mainly used for plant division propagation, but the defects of large seed consumption, low efficiency and long period are that the requirements of large-scale planting cannot be met, and the bletilla striata medicinal materials are listed as secondary protection plants by national key protection wild plant records (batch 2).
In order to prevent further damage of wild resources and guarantee market demands of bletilla striata, the bletilla striata seedling breeding is mainly carried out in a tissue culture mode at present. At present, the large-scale planting is carried out in regions such as southwest China, Yangtze river watershed and the like. However, the bletilla striata tissue culture seedling raising mode is high in production cost, large in labor consumption and relatively low in transplanting survival rate, and cannot meet the requirements of the medicinal material market on seedlings.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a bletilla direct seeding and seedling raising method by utilizing symbiotic bacteria, the bletilla symbiotic bacteria are added in the bletilla seed germination process after the bletilla symbiotic bacteria are subjected to separation culture, the direct seeding germination rate of bletilla seeds is greatly improved, a Huabao foliar fertilizer is sprayed in the later management process, the seedling growth speed is high, the production cost is low, the labor consumption is low, the cultured bletilla seedlings grow vigorously, the environmental adaptability is strong, and the transplanting survival rate is high.
In order to solve the technical problems, the invention adopts the technical scheme that: the bletilla direct seeding and seedling raising method utilizing symbiotic bacteria specifically comprises the following steps:
s1 obtaining bletilla striata symbiotic bacteria: culturing bletilla striata explants to obtain bletilla striata symbiotic bacteria strains for later use; s2 direct seeding and seedling raising of bletilla striata: seed mixing is carried out on the pretreated bletilla striata seeds and the bletilla striata symbiotic bacteria in the step S1, and then the seeds are sowed in a seedbed;
s3 sowing later-stage management: after the sowed seeds sprout out, fertilizing and hardening the seedlings, and then transplanting.
Through the technical scheme, the bletilla striata symbiotic bacteria are prepared, the bletilla striata symbiotic bacteria and bletilla striata seeds are mixed uniformly and then are subjected to direct seeding, and then seedling hardening treatment is performed, the bletilla striata symbiotic bacteria are added in the germination process of the bletilla striata seeds, so that the direct seeding germination rate of the bletilla striata seeds is up to 86%, and the Huabao foliar fertilizer is sprayed in the later management process, so that the seedling growth speed is high.
Further, the step S1 of obtaining bletilla striata symbiotic bacteria specifically includes the following steps:
s11 separation of bletilla striata symbiotic bacteria: sterilizing and disinfecting bletilla striata explants, cutting the bletilla striata explants into small blocks of 0.5-1 cm, inoculating the small blocks into a sterilized PDA culture medium, and culturing the small blocks in a constant-temperature incubator at 25 ℃ for 2-15 days;
s12 purification of bletilla striata symbiotic bacteria: after endophytes grow around the bletilla striata explants, picking hyphae on the edges of colonies, inoculating the hyphae to a new PDA culture medium, and culturing in a constant temperature incubator at 25 ℃ for 3-7 d; s13 cultivation of bletilla striata symbiotic bacteria: filling the prepared culture medium into culture bags, sterilizing for 1-1.5 h by high-temperature high-pressure steam, cooling, inoculating bletilla striata symbiotic bacteria strains in an aseptic environment, sealing the openings of the bags, transferring into a 25 ℃ culture room for culturing for 20-50 d, and allowing the culture bags to be full of mycelia for later use. The PDA medium composition in step S12 is the same as in step S11.
Further, the step S2 of directly seeding and raising the bletilla striata specifically includes the following steps:
s21 seedbed arrangement: ploughing and leveling the direct seeding ground, uniformly stirring peat soil and perlite according to a ratio of 8:2, paving 0.5-3 cm thick ground as a direct seeding matrix, and arranging waterlogging draining ditches every 0.8-1 m wide, wherein the depth is 8-12 cm;
s22 pretreatment of bletilla striata seeds: selecting mature and plump bletilla striata fruit pods, cutting the bletilla striata fruit pods, taking out all bletilla striata seeds, and then mixing the bletilla striata seeds with plant ash according to a ratio of 1: 50-1: 80 to obtain a bletilla seed plant ash mixture;
s23 seed dressing: dividing the bletilla striata symbiotic bacteria cultured in the step S1 into small blocks of 3-8 cm, and then fully and uniformly mixing the bletilla striata symbiotic bacteria with the bletilla striata seed plant ash mixture in the step S22 to obtain a bletilla striata seed and bletilla striata symbiotic bacteria mixture;
s24 sowing: sowing seeds with the length of 3-5 m according to each bletilla striata fruit pod2Uniformly sowing the mixed bletilla striata seeds and bletilla striata symbiotic bacteria on a seedbed, then spreading a layer of fine-grained soil of 0.3-0.5 cm, uniformly covering a layer of leaves above the fine-grained soil, and then spraying water by using a spraying pot, wherein the water spraying amount is 0.8-1.5L per square meter. The using amount of bletilla striata symbiotic bacteria mixed in each bletilla striata fruit pod is 1-2 Kg, and the bletilla striata symbiotic bacteria can effectively improve the self-growth of bletilla striata seedsGermination rate, however, promotes seedling growth, wherein soil with a coarse content in soil (i.e., a soil particle size greater than 0.075mm) of less than 50% is generally referred to as fine soil.
Further, the step S3 of post-seeding management specifically includes the following steps:
s31 film covering and scaffolding: after water is sprayed, covering a layer of mulching film on the seeds, and then building a simple shed above the mulching film by using a support and a plastic film;
s32 sowing later-stage management: when bletilla striata seeds germinate to seedlings of about 0.5cm, removing the film, spraying a Huabao foliar fertilizer every 10-15 days, wherein the fertilizing amount is 0.3-0.8L per square meter, controlling the temperature in the simple shed to be 18-28 ℃ and controlling the humidity to be 60-90%;
s33 seedling hardening treatment: when the seedlings grow to about 4cm, erecting and ventilating the two ends of the simple shed in the daytime, and covering the two ends of the simple shed at night; when the seedlings grow to about 5cm, removing the film of the simple shed in the daytime, covering the film at night and erecting the two ends of the film; transplanting the plantlets to a field for planting after the plantlets grow to about 7 cm. The simple shed is built by adopting the film and the small shed, so that moisture preservation and heat preservation can be effectively realized, and a favorable environment is provided for seed germination and seedling growth of bletilla striata; the gradual ventilation and the membrane removal in the seedling hardening process are used for enabling the seedlings to gradually adapt to the natural environment of the field and improving the transplanting survival rate, so that the transplanting survival rate is finally up to 99%.
Further, the bletilla striata explants used in step S1 are round bulbs, bletilla tubers and bletilla root hairs of the tissue culture germination of the bletilla striata seeds.
Further, the PDA culture medium used in the step S1 contains 200g/L of potatoes, 20g/L of sucrose and 20g/L of agar, and the pH is adjusted to 5-6.
Further, the step S11 of sterilizing the bletilla striata explant specifically comprises the steps of: the bletilla striata explants are cleaned by clear water, then disinfected by 75% alcohol for 15s, washed by sterile water for 2 times, then disinfected in 0.1% mercuric chloride for 3-5 min, and washed by sterile water for 5-7 times.
Further, the components of the culture medium in the step S13 are 62% of deciduous leaves of fagaceae, 8% of cottonseed hulls, 13.5% of broadleaf tree sawdust, 10% of rice bran or wheat bran, 1.5% of sucrose, 1.2% of gypsum, 0.8% of magnesium sulfate, 1.2% of monopotassium phosphate and 1.8% of urea, and a proper amount of water is added to fully and uniformly mix the components, so that the final water content is 65-70%.
Further, the foliar fertilizer for the Huabao used in the step S32 is 0.6g/L of Huabao No. 5 plus 0.2g/L of Huabao No. 2. The content ratio of nitrogen, phosphorus and potassium in the Huabao No. 5 is 30: 10: 10, the content ratio of nitrogen, phosphorus and potassium in Huabao 2 is 20: 20: 20, providing sufficient nutrition for the growth of the bletilla striata seedlings.
Compared with the prior art, the invention has the beneficial effects that: through germinating the in-process at the tuber of hyacinth bletilla seed and adding tuber of hyacinth bletilla symbiotic fungus, improved the natural direct seeding germination rate of tuber of hyacinth bletilla seed greatly, promote the healthy growth of tuber of hyacinth bletilla seedling, later stage management process spraying flower is precious foliar fertilizer, for tuber of hyacinth bletilla seedling growth provides sufficient nutrient, tuber of hyacinth bletilla seedling growth is fast, and the growth vigor is strong, and this method effectively reduces tuber of hyacinth bletilla seedling low in production cost, and the artificial consumption is few, and the tuber of hyacinth bletilla seedling growth of cultivateing is vigorous, and environmental adaptability is strong, and it is high to transplant the survival rate.
Detailed Description
The following examples further illustrate the present invention but are not to be construed as limiting the invention. Modifications and substitutions to methods, procedures, or conditions of the invention may be made without departing from the spirit of the invention.
Example 1: the bletilla direct seeding and seedling raising method utilizing symbiotic bacteria specifically comprises the following steps:
s1 obtaining bletilla striata symbiotic bacteria: culturing bletilla striata explants to obtain bletilla striata symbiotic bacteria strains for later use;
s11 separation of bletilla striata symbiotic bacteria: sterilizing and disinfecting bletilla striata explants, comprising the following steps: firstly, cleaning bletilla striata explants (including round bulbs, bletilla striata tubers and bletilla striata root hairs of tissue culture germination of bletilla striata seeds) with clear water, then disinfecting for 15s with 75% alcohol, washing for 2 times with sterile water, then disinfecting for 4min in 0.1% mercuric chloride, washing for 5 times with sterile water, cutting the disinfected bletilla striata explants into small blocks of 0.6cm, and inoculating the small blocks into a sterilized PDA culture medium, wherein the PDA culture medium contains 200g/L of potatoes, 20g/L of cane sugar, 20g/L of agar and 5-6 of pH modulation; then placing the mixture in a constant temperature incubator at 25 ℃ for culture for 7 d;
s12 purification of bletilla striata symbiotic bacteria: after endophytes grow around the bletilla striata explant, selecting hyphae at the edge of a bacterial colony, inoculating the hyphae to a new PDA culture medium, and culturing in a constant temperature incubator at 25 ℃ for 5 d;
s13 cultivation of bletilla striata symbiotic bacteria: filling the prepared culture medium into a culture bag, sterilizing for 1h by high-temperature high-pressure steam, cooling, inoculating strain of bletilla striata symbiotic bacteria in an aseptic environment, sealing the opening of the bag, transferring into a 25 ℃ culture chamber for culture for 30d, and allowing mycelia to grow to fill the culture bag for later use; wherein the components of the culture medium comprise 62 percent of deciduous leaves of fagaceae plants, 8 percent of cottonseed hulls, 13.5 percent of broadleaf tree sawdust, 10 percent of rice bran or wheat bran, 1.5 percent of cane sugar, 1.2 percent of gypsum, 0.8 percent of magnesium sulfate, 1.2 percent of monopotassium phosphate and 1.8 percent of urea, and a proper amount of water is added to fully and uniformly mix the components, and the final water content is 65-70 percent;
s2 direct seeding and seedling raising of bletilla striata: seed mixing is carried out on the pretreated bletilla striata seeds and the bletilla striata symbiotic bacteria in the step S1, and then the seeds are sowed in a seedbed;
s21 seedbed arrangement: ploughing and leveling the direct seeding ground, uniformly stirring peat soil and perlite according to a ratio of 8:2, paving 1.5cm thick ground as a direct seeding matrix, and arranging waterlogging draining ditches every 0.8m wide, wherein the depth is 10 cm;
s22 pretreatment of bletilla striata seeds: selecting mature and plump bletilla striata fruit pods, cutting the bletilla striata fruit pods, taking out all bletilla striata seeds, and then mixing the bletilla striata seeds with plant ash according to a ratio of 1: 50 to obtain a bletilla seed plant ash mixture;
s23 seed dressing: dividing the bletilla striata symbiotic bacteria cultured in the step S1 into small blocks of 3-5 cm, and fully and uniformly mixing the bletilla striata symbiotic bacteria with the bletilla striata seed plant ash mixture in the step S22 to obtain a bletilla striata seed and bletilla striata symbiotic bacteria mixture; the using amount of bletilla striata symbiotic bacteria mixed in each bletilla striata fruit pod is 1.5Kg, and the bletilla striata symbiotic bacteria can effectively improve the natural germination rate of bletilla striata seeds and promote the growth of seedlings;
s24 sowing: sowing seeds at a distance of 3.5m per each capsule of bletilla striata2Uniformly sowing the mixed bletilla striata seeds and bletilla striata symbiotic bacteria on a seedbed, then spreading a layer of fine soil of 0.3cm, uniformly covering a layer of leaves above the fine soil, and then spraying water by using a spraying pot, wherein the water spraying amount is 1.2L per square meter;
s3 sowing later-stage management: after the sowed seeds sprout and emit seedlings, fertilizing and hardening the seedlings, and then transplanting the seedlings;
s31 film covering and scaffolding: after water is sprayed, covering a layer of mulching film on the seeds, and then building a simple shed above the mulching film by using a support and a plastic film; the simple shed is built by adopting the film and the small shed, so that moisture preservation and heat preservation can be effectively realized, and a favorable environment is provided for seed germination and seedling growth of bletilla striata;
s32 sowing later-stage management: when bletilla striata seeds germinate to seedlings of about 0.5cm, uncovering the film, spraying a Huabao foliar fertilizer every 10 days, wherein the fertilizing amount is 0.6L per square meter, controlling the temperature in the simple shed to be 20-22 ℃ and controlling the humidity to be 60-70%; the foliar fertilizer for the Huabao used in the step S32 is 0.6g/L of Huabao No. 5 plus 0.2g/L of Huabao No. 2; the content ratio of nitrogen, phosphorus and potassium in the Huabao No. 5 is 30: 10: 10, the content ratio of nitrogen, phosphorus and potassium in Huabao 2 is 20: 20: 20, providing sufficient nutrition for the growth of the bletilla striata seedlings;
s33 seedling hardening treatment: when the seedlings grow to about 4cm, erecting and ventilating the two ends of the simple shed in the daytime, and covering the two ends of the simple shed at night; when the seedlings grow to about 5cm, removing the film of the simple shed in the daytime, covering the film at night and erecting the two ends of the film; transplanting the plantlets to a field for planting when the plantlets grow to about 7 cm; the gradual ventilation and the membrane removal in the seedling hardening process are used for enabling the seedlings to gradually adapt to the natural environment of the field and improving the transplanting survival rate. The bletilla striata seed germination rate of the embodiment 1 is 86%, and the seedling transplanting survival rate is 98%.
Example 2: the bletilla direct seeding and seedling raising method utilizing symbiotic bacteria specifically comprises the following steps:
s1 obtaining bletilla striata symbiotic bacteria: culturing bletilla striata explants to obtain bletilla striata symbiotic bacteria strains for later use;
s11 separation of bletilla striata symbiotic bacteria: sterilizing and disinfecting bletilla striata explants, comprising the following steps: firstly, cleaning bletilla striata explants (including round bulbs, bletilla striata tubers and bletilla striata root hairs of tissue culture germination of bletilla striata seeds) with clear water, then disinfecting for 15s with 75% alcohol, washing for 2 times with sterile water, then disinfecting for 5min in 0.1% mercuric chloride, washing for 5 times with sterile water, cutting the sterilized bletilla striata explants into small blocks of 0.8cm, and inoculating the small blocks into sterilized PDA culture medium, wherein the used PDA culture medium contains 200g/L of potatoes, 20g/L of cane sugar, 20g/L of agar and 5-6 of pH modulation; then placing the mixture in a constant temperature incubator at 25 ℃ for culturing for 10 d;
s12 purification of bletilla striata symbiotic bacteria: after endophytes grow around the bletilla striata explant, selecting hyphae at the edge of a bacterial colony, inoculating the hyphae to a new PDA culture medium, and culturing in a constant-temperature incubator at 25 ℃ for 7 d;
s13 cultivation of bletilla striata symbiotic bacteria: filling the prepared culture medium into culture bags, sterilizing for 1.2h with high-temperature high-pressure steam, cooling, inoculating strain of symbiotic bacteria of bletilla striata in aseptic environment, sealing bag opening, transferring into 25 deg.C culture chamber, culturing for 50d, and allowing mycelia to grow over the culture bags; wherein the components of the culture medium comprise 62 percent of deciduous leaves of fagaceae plants, 8 percent of cottonseed hulls, 13.5 percent of broadleaf tree sawdust, 10 percent of rice bran or wheat bran, 1.5 percent of cane sugar, 1.2 percent of gypsum, 0.8 percent of magnesium sulfate, 1.2 percent of monopotassium phosphate and 1.8 percent of urea, and a proper amount of water is added to fully and uniformly mix the components, and the final water content is 65-70 percent;
s2 direct seeding and seedling raising of bletilla striata: seed mixing is carried out on the pretreated bletilla striata seeds and the bletilla striata symbiotic bacteria in the step S1, and then the seeds are sowed in a seedbed;
s21 seedbed arrangement: ploughing and leveling the direct seeding ground, uniformly stirring peat soil and perlite according to a ratio of 8:2, paving 2cm thick ground as a direct seeding matrix, and arranging waterlogging draining ditches every 0.9m wide to 8cm deep;
s22 pretreatment of bletilla striata seeds: selecting mature and plump bletilla striata fruit pods, cutting the bletilla striata fruit pods, taking out all bletilla striata seeds, and then mixing the bletilla striata seeds with plant ash according to a ratio of 1: 70 to obtain a bletilla seed plant ash mixture;
s23 seed dressing: dividing the bletilla striata symbiotic bacteria cultured in the step S1 into small blocks of 5-6 cm, and fully and uniformly mixing the bletilla striata symbiotic bacteria with the bletilla striata seed plant ash mixture in the step S22 to obtain a bletilla striata seed and bletilla striata symbiotic bacteria mixture; the using amount of bletilla striata symbiotic bacteria mixed in each bletilla striata fruit pod is 1.2Kg, and the bletilla striata symbiotic bacteria can effectively improve the natural germination rate of bletilla striata seeds and promote the growth of seedlings;
s24 sowing: sowing seeds of 4.5m per each bletilla striata fruit pod2Uniformly sowing the mixed bletilla striata seeds and bletilla striata symbiotic bacteria on a seedbed, then spreading a layer of fine soil of 0.4cm, uniformly covering a layer of leaves above the fine soil, and then spraying water by using a spraying pot, wherein the water spraying amount is 1.5L per square meter;
s3 sowing later-stage management: after the sowed seeds sprout and emit seedlings, fertilizing and hardening the seedlings, and then transplanting the seedlings;
s31 film covering and scaffolding: after water is sprayed, covering a layer of mulching film on the seeds, and then building a simple shed above the mulching film by using a support and a plastic film; the simple shed is built by adopting the film and the small shed, so that moisture preservation and heat preservation can be effectively realized, and a favorable environment is provided for seed germination and seedling growth of bletilla striata;
s32 sowing later-stage management: when bletilla striata seeds germinate to seedlings of about 0.5cm, uncovering the film, spraying a Huabao foliar fertilizer every 12 days, wherein the fertilizing amount is 0.5L per square meter, controlling the temperature in the simple shed to be 22-25 ℃ and controlling the humidity to be 65-80%; the foliar fertilizer for the Huabao used in the step S32 is 0.6g/L of Huabao No. 5 plus 0.2g/L of Huabao No. 2; the content ratio of nitrogen, phosphorus and potassium in the Huabao No. 5 is 30: 10: 10, the content ratio of nitrogen, phosphorus and potassium in Huabao 2 is 20: 20: 20, providing sufficient nutrition for the growth of the bletilla striata seedlings;
s33 seedling hardening treatment: when the seedlings grow to about 4cm, erecting and ventilating the two ends of the simple shed in the daytime, and covering the two ends of the simple shed at night; when the seedlings grow to about 5cm, removing the film of the simple shed in the daytime, covering the film at night and erecting the two ends of the film; transplanting the plantlets to a field for planting when the plantlets grow to about 7 cm; the gradual ventilation and the membrane removal in the seedling hardening process are used for enabling the seedlings to gradually adapt to the natural environment of the field and improving the transplanting survival rate. The bletilla striata seed germination rate of the embodiment 2 is 83%, and the seedling transplanting survival rate is 96%.
Example 3: the bletilla direct seeding and seedling raising method utilizing symbiotic bacteria specifically comprises the following steps:
s1 obtaining bletilla striata symbiotic bacteria: culturing bletilla striata explants to obtain bletilla striata symbiotic bacteria strains for later use;
s11 separation of bletilla striata symbiotic bacteria: sterilizing and disinfecting bletilla striata explants, comprising the following steps: firstly, cleaning bletilla striata explants (including round bulbs, bletilla striata tubers and bletilla striata root hairs of tissue culture germination of bletilla striata seeds) with clear water, then disinfecting for 15s with 75% alcohol, washing for 2 times with sterile water, then disinfecting for 3.5min in 0.1% mercuric chloride, washing for 6 times with sterile water, cutting the sterilized bletilla striata explants into small blocks of 1cm, and inoculating the small blocks into sterilized PDA culture medium, wherein the used PDA culture medium contains 200g/L of potatoes, 20g/L of cane sugar, 20g/L of agar and 5-6 of pH modulation; then placing the mixture in a constant temperature incubator at 25 ℃ for culture for 13 d;
s12 purification of bletilla striata symbiotic bacteria: after endophytes grow around the bletilla striata explant, selecting hyphae at the edge of a bacterial colony, inoculating the hyphae to a new PDA culture medium, and culturing in a constant temperature incubator at 25 ℃ for 6 d;
s13 cultivation of bletilla striata symbiotic bacteria: filling the prepared culture medium into culture bags, sterilizing for 1.5h with high-temperature high-pressure steam, cooling, inoculating strain of symbiotic bacteria of bletilla striata in aseptic environment, sealing bag opening, transferring into 25 deg.C culture chamber, culturing for 45d, and allowing mycelia to grow over the culture bags; wherein the components of the culture medium comprise 62 percent of deciduous leaves of fagaceae plants, 8 percent of cottonseed hulls, 13.5 percent of broadleaf tree sawdust, 10 percent of rice bran or wheat bran, 1.5 percent of cane sugar, 1.2 percent of gypsum, 0.8 percent of magnesium sulfate, 1.2 percent of monopotassium phosphate and 1.8 percent of urea, and a proper amount of water is added to fully and uniformly mix the components, and the final water content is 65-70 percent;
s2 direct seeding and seedling raising of bletilla striata: seed mixing is carried out on the pretreated bletilla striata seeds and the bletilla striata symbiotic bacteria in the step S1, and then the seeds are sowed in a seedbed;
s21 seedbed arrangement: ploughing and leveling the direct seeding ground, uniformly stirring peat soil and perlite according to a ratio of 8:2, paving 2.8cm thick ground as a direct seeding matrix, and arranging waterlogging draining ditches every 1m wide to a depth of 9 cm;
s22 pretreatment of bletilla striata seeds: selecting mature and plump bletilla striata fruit pods, cutting the bletilla striata fruit pods, taking out all bletilla striata seeds, and then mixing the bletilla striata seeds with plant ash according to a ratio of 1: 80 to obtain a bletilla seed plant ash mixture;
s23 seed dressing: dividing the bletilla striata symbiotic bacteria cultured in the step S1 into small blocks of 6-8 cm, and fully and uniformly mixing the bletilla striata symbiotic bacteria with the bletilla striata seed plant ash mixture in the step S22 to obtain a bletilla striata seed and bletilla striata symbiotic bacteria mixture; the using amount of bletilla striata symbiotic bacteria mixed in each bletilla striata fruit pod is 1.8Kg, and the bletilla striata symbiotic bacteria can effectively improve the natural germination rate of bletilla striata seeds and promote the growth of seedlings;
s24 sowing: sowing seeds at a distance of 4m per each bletilla striata pod2Uniformly sowing the mixed bletilla striata seeds and bletilla striata symbiotic bacteria on a seedbed, then spreading a layer of fine soil of 0.5cm, uniformly covering a layer of leaves above the fine soil, and then spraying water by using a spraying pot, wherein the water spraying amount is 1.0L per square meter;
s3 sowing later-stage management: after the sowed seeds sprout and emit seedlings, fertilizing and hardening the seedlings, and then transplanting the seedlings;
s31 film covering and scaffolding: after water is sprayed, covering a layer of mulching film on the seeds, and then building a simple shed above the mulching film by using a support and a plastic film; the simple shed is built by adopting the film and the small shed, so that moisture preservation and heat preservation can be effectively realized, and a favorable environment is provided for seed germination and seedling growth of bletilla striata;
s32 sowing later-stage management: when bletilla striata seeds germinate to seedlings of about 0.5cm, uncovering the film, spraying a Huabao foliar fertilizer every 14 days, wherein the fertilizing amount is 0.8L per square meter, controlling the temperature in the simple shed to be 24-26 ℃, and controlling the humidity to be 75-90%; the foliar fertilizer for the Huabao used in the step S32 is 0.6g/L of Huabao No. 5 plus 0.2g/L of Huabao No. 2; the content ratio of nitrogen, phosphorus and potassium in the Huabao No. 5 is 30: 10: 10, the content ratio of nitrogen, phosphorus and potassium in Huabao 2 is 20: 20: 20, providing sufficient nutrition for the growth of the bletilla striata seedlings;
s33 seedling hardening treatment: when the seedlings grow to about 4cm, erecting and ventilating the two ends of the simple shed in the daytime, and covering the two ends of the simple shed at night; when the seedlings grow to about 5cm, removing the film of the simple shed in the daytime, covering the film at night and erecting the two ends of the film; transplanting the plantlets to a field for planting when the plantlets grow to about 7 cm; the gradual ventilation and the membrane removal in the seedling hardening process are used for enabling the seedlings to gradually adapt to the natural environment of the field and improving the transplanting survival rate. The bletilla striata seed germination rate of the embodiment 3 is 84%, and the seedling transplanting survival rate is 99%.
Finally, it is also noted that the above-mentioned lists merely illustrate a few specific embodiments of the invention. Obviously, the present invention is not limited to the above embodiments, but may be modified in many ways; all modifications which can be derived or suggested by a person skilled in the art from the disclosure of the present invention are to be considered within the scope of the invention.
Claims (9)
1. A bletilla direct seeding and seedling raising method utilizing symbiotic bacteria is characterized by comprising the following steps:
s1 obtaining bletilla striata symbiotic bacteria: culturing bletilla striata explants to obtain bletilla striata symbiotic bacteria strains for later use;
s2 direct seeding and seedling raising of bletilla striata: seed mixing is carried out on the pretreated bletilla striata seeds and the bletilla striata symbiotic bacteria in the step S1, and then the seeds are sowed in a seedbed;
s3 sowing later-stage management: after the sowed seeds sprout out, fertilizing and hardening the seedlings, and then transplanting.
2. The bletilla striata direct seeding and seedling raising method by utilizing symbiotic bacteria according to claim 1, wherein the step S1 of obtaining bletilla striata symbiotic bacteria specifically comprises the following steps:
s11 separation of bletilla striata symbiotic bacteria: sterilizing and disinfecting bletilla striata explants, cutting the bletilla striata explants into small blocks of 0.5-1 cm, inoculating the small blocks into a sterilized PDA culture medium, and culturing the small blocks in a constant-temperature incubator at 25 ℃ for 2-15 days;
s12 purification of bletilla striata symbiotic bacteria: after endophytes grow around the bletilla striata explants, picking hyphae on the edges of colonies, inoculating the hyphae to a new PDA culture medium, and culturing in a constant temperature incubator at 25 ℃ for 3-7 d;
s13 cultivation of bletilla striata symbiotic bacteria: filling the prepared culture medium into culture bags, sterilizing for 1-1.5 h by high-temperature high-pressure steam, cooling, inoculating bletilla striata symbiotic bacteria strains in an aseptic environment, sealing the openings of the bags, transferring into a 25 ℃ culture room for culturing for 20-50 d, and allowing the culture bags to be full of mycelia for later use.
3. The bletilla direct seeding and seedling raising method by using symbiotic bacteria according to claim 2, wherein the step S2 comprises the following steps:
s21 seedbed arrangement: ploughing and leveling the direct seeding ground, uniformly stirring peat soil and perlite according to a ratio of 8:2, paving 0.5-3 cm thick ground as a direct seeding matrix, and arranging waterlogging draining ditches every 0.8-1 m wide, wherein the depth is 8-12 cm;
s22 pretreatment of bletilla striata seeds: selecting mature and plump bletilla striata fruit pods, cutting the bletilla striata fruit pods, taking out all bletilla striata seeds, and then mixing the bletilla striata seeds with plant ash according to a ratio of 1: 50-1: 80 to obtain a bletilla seed plant ash mixture;
s23 seed dressing: dividing the bletilla striata symbiotic bacteria cultured in the step S1 into small blocks of 3-8 cm, and then fully and uniformly mixing the bletilla striata symbiotic bacteria with the bletilla striata seed plant ash mixture in the step S22 to obtain a bletilla striata seed and bletilla striata symbiotic bacteria mixture;
s24 sowing: sowing seeds with the length of 3-5 m according to each bletilla striata fruit pod2Uniformly sowing the mixed bletilla striata seeds and bletilla striata symbiotic bacteria on a seedbed, then spreading a layer of fine-grained soil of 0.3-0.5 cm, uniformly covering a layer of leaves above the fine-grained soil, and then spraying water by using a spraying pot, wherein the water spraying amount is 0.8-1.5L per square meter.
4. The bletilla direct seeding and seedling raising method by using symbiotic bacteria according to claim 3, wherein the step S3 of late sowing management specifically comprises the following steps:
s31 film covering and scaffolding: after water is sprayed, covering a layer of mulching film on the seeds, and then building a simple shed above the mulching film by using a support and a plastic film;
s32 sowing later-stage management: when bletilla striata seeds germinate to seedlings of about 0.5cm, removing the film, spraying a Huabao foliar fertilizer every 10-15 days, wherein the fertilizing amount is 0.3-0.8L per square meter, controlling the temperature in the simple shed to be 18-28 ℃ and controlling the humidity to be 60-90%;
s33 seedling hardening treatment: when the seedlings grow to about 4cm, erecting and ventilating the two ends of the simple shed in the daytime, and covering the two ends of the simple shed at night; when the seedlings grow to about 5cm, removing the film of the simple shed in the daytime, covering the film at night and erecting the two ends of the film; transplanting the plantlets to a field for planting after the plantlets grow to about 7 cm.
5. The bletilla direct seeding and seedling raising method by using symbiotic bacteria according to claim 4, wherein the bletilla explants used in step S1 are bletilla bulbils, bletilla tubers and bletilla root hairs of bletilla seed tissue culture germination.
6. The bletilla striata direct seeding and seedling raising method by utilizing symbiotic bacteria according to claim 4, wherein the PDA culture medium used in the step S1 contains 200g/L of potatoes, 20g/L of cane sugar and 20g/L of agar, and the pH is adjusted to 5-6.
7. The bletilla direct seeding and seedling raising method by using symbiotic bacteria according to claim 4, wherein the specific steps of sterilizing and disinfecting the bletilla explants in the step S11 are as follows: the bletilla striata explants are cleaned by clear water, then disinfected by 75% alcohol for 15s, washed by sterile water for 2 times, then disinfected in 0.1% mercuric chloride for 3-5 min, and washed by sterile water for 5-7 times.
8. The bletilla direct seeding and seedling raising method by using symbiotic bacteria according to claim 4, wherein the components of the culture medium in the step S13 are 62% of deciduous plant deciduous leaves, 8% of cottonseed hulls, 13.5% of broadleaf tree sawdust, 10% of rice bran or wheat bran, 1.5% of sucrose, 1.2% of gypsum, 0.8% of magnesium sulfate, 1.2% of monopotassium phosphate and 1.8% of urea, and a proper amount of water is added to fully and uniformly stir the components, so that the final water content is 65-70%.
9. The bletilla striata direct seeding and seedling raising method by utilizing symbiotic bacteria according to claim 4, wherein the foliar fertilizer for the Huabao in the step S32 is 0.6g/L of Huabao No. 5 plus 0.2g/L of Huabao No. 2.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110471049.5A CN113207590A (en) | 2021-04-29 | 2021-04-29 | Bletilla direct seeding and seedling raising method utilizing symbiotic bacteria |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110471049.5A CN113207590A (en) | 2021-04-29 | 2021-04-29 | Bletilla direct seeding and seedling raising method utilizing symbiotic bacteria |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113207590A true CN113207590A (en) | 2021-08-06 |
Family
ID=77089923
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110471049.5A Pending CN113207590A (en) | 2021-04-29 | 2021-04-29 | Bletilla direct seeding and seedling raising method utilizing symbiotic bacteria |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113207590A (en) |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101914457A (en) * | 2008-04-24 | 2010-12-15 | 中国医学科学院药用植物研究所 | Fungus for promoting germination of bletilla striata seeds |
| CN105103924A (en) * | 2015-09-25 | 2015-12-02 | 马龙县兴裕农业生物科技开发有限公司 | High-yield cultivation method for pollution-free bletilla striata |
| CN105103714A (en) * | 2015-08-25 | 2015-12-02 | 长江大学 | Manual germination accelerating method and seedling raising method of Bletilla sfriata (Thunb.)Reiehb.f. |
| CN105519350A (en) * | 2015-12-14 | 2016-04-27 | 广西壮族自治区药用植物园 | Method for rapidly producing Bletilla striata germchit |
| CN105659979A (en) * | 2016-01-18 | 2016-06-15 | 云南绿辰生物科技开发有限公司 | Method for cultivating symbiotic bacteria of Bletilla striata |
| CN106797791A (en) * | 2017-01-24 | 2017-06-06 | 贵州省武陵山药用植物白芨开发有限公司 | The type of seeding of the bletilla striata and its cultural method |
| CN108371003A (en) * | 2016-10-26 | 2018-08-07 | 贵州发来地农业科技有限公司 | A kind of breeding method of bletilla striata seeds |
| CN108371008A (en) * | 2016-10-27 | 2018-08-07 | 贵州发来地农业科技有限公司 | A kind of implantation methods of the good bletilla striata of quality |
| CN110122210A (en) * | 2019-06-18 | 2019-08-16 | 陇南市农业科学研究所 | A kind of bletilla seed sowing efficient cultivation method |
| CN110338047A (en) * | 2019-06-28 | 2019-10-18 | 长江大学 | A kind of bletilla striata seeds imitate wild efficient direct sowing and seedling method |
-
2021
- 2021-04-29 CN CN202110471049.5A patent/CN113207590A/en active Pending
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101914457A (en) * | 2008-04-24 | 2010-12-15 | 中国医学科学院药用植物研究所 | Fungus for promoting germination of bletilla striata seeds |
| CN105103714A (en) * | 2015-08-25 | 2015-12-02 | 长江大学 | Manual germination accelerating method and seedling raising method of Bletilla sfriata (Thunb.)Reiehb.f. |
| CN105103924A (en) * | 2015-09-25 | 2015-12-02 | 马龙县兴裕农业生物科技开发有限公司 | High-yield cultivation method for pollution-free bletilla striata |
| CN105519350A (en) * | 2015-12-14 | 2016-04-27 | 广西壮族自治区药用植物园 | Method for rapidly producing Bletilla striata germchit |
| CN105659979A (en) * | 2016-01-18 | 2016-06-15 | 云南绿辰生物科技开发有限公司 | Method for cultivating symbiotic bacteria of Bletilla striata |
| CN108371003A (en) * | 2016-10-26 | 2018-08-07 | 贵州发来地农业科技有限公司 | A kind of breeding method of bletilla striata seeds |
| CN108371008A (en) * | 2016-10-27 | 2018-08-07 | 贵州发来地农业科技有限公司 | A kind of implantation methods of the good bletilla striata of quality |
| CN106797791A (en) * | 2017-01-24 | 2017-06-06 | 贵州省武陵山药用植物白芨开发有限公司 | The type of seeding of the bletilla striata and its cultural method |
| CN110122210A (en) * | 2019-06-18 | 2019-08-16 | 陇南市农业科学研究所 | A kind of bletilla seed sowing efficient cultivation method |
| CN110338047A (en) * | 2019-06-28 | 2019-10-18 | 长江大学 | A kind of bletilla striata seeds imitate wild efficient direct sowing and seedling method |
Non-Patent Citations (1)
| Title |
|---|
| 封海东等: "大棚内白芨种子直播育苗技术", 《湖北农业科学》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102356746B (en) | Technology for cultivating organic fruit and vegetable matrix | |
| CN102599026B (en) | Tilia miqueliana seedling raising method | |
| CN103598098B (en) | Pachyrhizua angulatus tissue culturing fast seedling-cultivating method | |
| CN103828717A (en) | Rapid propagating and seedling culturing method by virtue of Bletilla tubers | |
| CN102648696B (en) | Method for producing vegetable cultivation substrate and seedling-raising substrate by using medlar branches | |
| CN103416187A (en) | Towel gourd cultivation method | |
| CN103460954A (en) | Early-stage culture method for white gourd | |
| CN102301874A (en) | Method for culturing seedlings of Phoebe bournei by light soil type mixed matrix | |
| CN103947433A (en) | One-step seedling forming method of bletilla striata tissue culture corm and substrate adopted by one-step seedling forming method | |
| CN108812116A (en) | A kind of composite stereo ecology method for interplanting cultivation of capsicum, sponge gourd and water spinach | |
| CN103694039B (en) | The light cultivation matrix of Hubei Province, a kind of river Camellia fraternal breeding and container seedling culture method thereof | |
| CN101697703B (en) | Method for bickiron inoculation of buds symbiotic to root fungi for culturing seedlings of Chinese chestnut | |
| CN104855445A (en) | Pomegranate seed treatment solution | |
| CN112602397A (en) | Soil biological disinfection method for vegetable greenhouse in continuous cropping facility | |
| CN109496724B (en) | Efficient planting method for emilia sonchifolia | |
| CN105900723A (en) | Panax japonicus var.major seedling method and culture method | |
| CN108094031A (en) | A kind of baeckea frutescen container seedling culture method | |
| CN104855216A (en) | Seedling culture method for preventing pomegranate seedling root rotting through symbiotic edible fungi | |
| CN106888948A (en) | A kind of seed propagation method of first hat rattan | |
| CN105145257A (en) | High-efficiency paraphlomis javanica var.coronata forest building method | |
| CN104429442B (en) | A kind of implantation methods of the twigs of the chaste tree | |
| CN114853515B (en) | Preparation method of organic nutrient soil | |
| CN104885732A (en) | Seedling breeding method for preventing root rot of pomegranate seedlings | |
| CN104926429A (en) | Seed treating fluid for preventing pomegranate seedlings from rotted roots | |
| CN100499997C (en) | Ainsliaea fragrans champ breeding method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |