CN116649216A - Pure bletilla striata three-fork tissue culture domesticated seedling raising method - Google Patents
Pure bletilla striata three-fork tissue culture domesticated seedling raising method Download PDFInfo
- Publication number
- CN116649216A CN116649216A CN202310748215.0A CN202310748215A CN116649216A CN 116649216 A CN116649216 A CN 116649216A CN 202310748215 A CN202310748215 A CN 202310748215A CN 116649216 A CN116649216 A CN 116649216A
- Authority
- CN
- China
- Prior art keywords
- culture
- bletilla striata
- domestication
- seedlings
- tissue culture
- 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
- 241001313857 Bletilla striata Species 0.000 title claims abstract description 53
- 238000000034 method Methods 0.000 title claims abstract description 39
- 239000002775 capsule Substances 0.000 claims abstract description 47
- 238000009331 sowing Methods 0.000 claims abstract description 34
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 31
- 239000011159 matrix material Substances 0.000 claims abstract description 27
- 238000012258 culturing Methods 0.000 claims abstract description 19
- 238000005728 strengthening Methods 0.000 claims abstract description 18
- 241001494479 Pecora Species 0.000 claims abstract description 13
- 210000003608 fece Anatomy 0.000 claims abstract description 13
- 239000010871 livestock manure Substances 0.000 claims abstract description 13
- 241001313855 Bletilla Species 0.000 claims abstract description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 11
- RCTYPNKXASFOBE-UHFFFAOYSA-M chloromercury Chemical compound [Hg]Cl RCTYPNKXASFOBE-UHFFFAOYSA-M 0.000 claims abstract description 9
- 239000005802 Mancozeb Substances 0.000 claims abstract description 8
- 239000007864 aqueous solution Substances 0.000 claims abstract description 8
- 238000002360 preparation method Methods 0.000 claims abstract description 5
- 239000001963 growth medium Substances 0.000 claims description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 22
- 230000001954 sterilising effect Effects 0.000 claims description 21
- 238000005507 spraying Methods 0.000 claims description 12
- 238000005286 illumination Methods 0.000 claims description 10
- 230000035784 germination Effects 0.000 claims description 8
- 238000002791 soaking Methods 0.000 claims description 8
- 238000002156 mixing Methods 0.000 claims description 7
- 239000002689 soil Substances 0.000 claims description 7
- 229920001817 Agar Polymers 0.000 claims description 6
- 235000013162 Cocos nucifera Nutrition 0.000 claims description 6
- 244000060011 Cocos nucifera Species 0.000 claims description 6
- 229930191978 Gibberellin Natural products 0.000 claims description 6
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 6
- 229930006000 Sucrose Natural products 0.000 claims description 6
- 239000008272 agar Substances 0.000 claims description 6
- 238000005520 cutting process Methods 0.000 claims description 6
- 239000003085 diluting agent Substances 0.000 claims description 6
- IXORZMNAPKEEDV-UHFFFAOYSA-N gibberellic acid GA3 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(C=CC3O)C1C3(C)C(=O)O2 IXORZMNAPKEEDV-UHFFFAOYSA-N 0.000 claims description 6
- 239000003448 gibberellin Substances 0.000 claims description 6
- 235000013575 mashed potatoes Nutrition 0.000 claims description 6
- 239000003415 peat Substances 0.000 claims description 6
- 239000008223 sterile water Substances 0.000 claims description 6
- 238000010899 nucleation Methods 0.000 claims description 5
- 230000012010 growth Effects 0.000 claims description 4
- 238000011010 flushing procedure Methods 0.000 claims description 3
- 238000011081 inoculation Methods 0.000 claims description 3
- 239000012466 permeate Substances 0.000 claims description 3
- 239000002352 surface water Substances 0.000 claims description 3
- 238000004140 cleaning Methods 0.000 claims description 2
- 238000000354 decomposition reaction Methods 0.000 claims description 2
- 239000003595 mist Substances 0.000 claims description 2
- 230000007480 spreading Effects 0.000 claims description 2
- 238000003892 spreading Methods 0.000 claims description 2
- 230000004083 survival effect Effects 0.000 abstract description 13
- 230000028446 budding cell bud growth Effects 0.000 abstract description 2
- 230000006872 improvement Effects 0.000 description 8
- 230000008901 benefit Effects 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 229960002523 mercuric chloride Drugs 0.000 description 2
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 description 2
- 208000034507 Haematemesis Diseases 0.000 description 1
- 208000000616 Hemoptysis Diseases 0.000 description 1
- 206010040849 Skin fissures Diseases 0.000 description 1
- 206010053476 Traumatic haemorrhage Diseases 0.000 description 1
- 235000019606 astringent taste Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 235000019658 bitter taste Nutrition 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 235000019605 sweet taste sensations Nutrition 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 238000009736 wetting Methods 0.000 description 1
Classifications
-
- 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
-
- 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
-
- 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
- A01G24/22—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material containing plant material
- A01G24/25—Dry fruit hulls or husks, e.g. chaff or coir
-
- 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
- A01H4/002—Culture media for tissue culture
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
- Y02P60/21—Dinitrogen oxide [N2O], e.g. using aquaponics, hydroponics or efficiency measures
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Environmental Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Cultivation Of Plants (AREA)
Abstract
The application discloses a pure bletilla striata three-fork tissue culture domestication seedling raising method, which comprises the steps of carrying out deep disinfection treatment by using alcohol and mercury chloride, sowing and culturing after seed taking, carrying out rooting and seedling strengthening culture of seedlings after seed taking, carrying out mixed preparation of domestication matrixes, and carrying out domestication culture of tissue culture seedlings; according to the application, the problem of low bud growth rate after sowing due to incomplete disinfection is greatly reduced by carrying out deep disinfection by alcohol and mercury chloride, meanwhile, the root is soaked by using the mancozeb aqueous solution before rooting and transferring, so that the rooting survival rate is greatly increased, and the survival rate of the domesticated tissue culture seedlings of the trigeminal bletilla is further improved by adding the decomposed capsule shells and sheep manure into the domestication matrix, and the yield of artificial planting of the trigeminal bletilla is comprehensively increased.
Description
Technical Field
The application relates to the technical field of trigeminal bletilla striata breeding, in particular to a pure trigeminal tissue culture domestication seedling raising method for bletilla striata.
Background
The bletilla striata is a traditional Chinese medicine name, tubers are picked up when stems and leaves of the bletilla striata wither for 9-10 months each year, fibrous roots are removed, the bletilla striata is cleaned, boiled in boiling water until no white core exists, and then the bletilla striata is dried to be semi-dry, the outer skin is removed, and the bletilla striata can be used as a medicine after being dried in the sun; it is used for treating hemoptysis, hematemesis, traumatic hemorrhage, pyocutaneous disease, toxic swelling and chapped skin due to its bitter, sweet and astringent taste and slightly cold nature.
Along with the use of the bletilla striata as a medicine, the quantity of the wild bletilla striata is rapidly reduced at present, most of the bletilla striata as a medicine is planted artificially at present, and the artificial planting of the trigeminal bletilla striata in the bletilla striata needs domestication and seedling raising treatment;
the traditional three-fork bletilla domestication seedling raising method is characterized in that common bletilla domestication seedling raising operation is adopted for breeding, simple disinfection is adopted for seed taking and utilizing a culture medium for seedling raising, domestication planting is adopted, the germination rate is low during domestication seedling raising caused by incomplete disinfection, the survival rate of subsequent culture is seriously influenced, the yield of artificial planting of the three-fork bletilla is seriously influenced, meanwhile, the three-fork bletilla domestication culture is carried out by using common nutrient soil with simple components, and the method is also one of important reasons for not improving the yield of the three-fork bletilla, so that the application provides a pure three-fork tissue culture domestication seedling raising method for the pure bletilla, and solves the problems in the prior art.
Disclosure of Invention
Aiming at the problems, the application aims to provide the pure bletilla striata three-fork tissue culture domestication seedling raising method, which is used for sowing and sprouting after deep disinfection by alcohol and mercury chloride, so that the problem of low sprouting rate after sowing caused by incomplete disinfection is greatly reduced, meanwhile, the root is soaked by using a mancozeb aqueous solution before root transfer, the rooting survival rate is greatly increased, and the rotten capsule shells and sheep manure are added into a domestication matrix, so that the survival rate of the three-fork bletilla striata domestication tissue culture seedling is further improved, and the yield of artificial planting of the three-fork bletilla striata is comprehensively increased.
In order to achieve the purpose of the application, the application is realized by the following technical scheme: a pure bletilla striata three-fork tissue culture domestication seedling raising method comprises the following steps:
the method comprises the steps of firstly, deeply sterilizing, sterilizing the mature and uncracked capsule of the bletilla striata by using alcohol and 0.1% mercury chloride under a sterile condition, and repeatedly flushing the capsule with sterile water for five times to obtain a sterilized capsule;
culturing after seed taking, namely sucking the surface water of the cleaned disinfection capsules by using sterile filter paper, cutting the capsules by using a disinfection scalpel, uniformly shaking off the internal seeds by using a sterile forceps, soaking the seeds in a germination accelerator diluent for 5-8 hours, fishing out the seeds, draining the seeds, and uniformly scattering the seeds on a sowing culture medium for sowing culture;
step three, rooting and seedling strengthening culture, namely, after sowing and culturing for two months, soaking the trigeminal bletilla striata seedlings which induce sprouting in a mancozeb aqueous solution, transferring the soaked trigeminal bletilla striata seedlings to a rooting and seedling strengthening culture medium, and performing seedling strengthening culture on the tissue culture seedlings;
step four, preparing an domestication matrix, namely crushing the capsule shells obtained by cutting and seed taking in the step two, and then uniformly mixing the capsule shells with coconut coir, peat soil and decomposed sheep manure to obtain the domestication matrix;
fifthly, domesticating and culturing, namely taking out the bottle when the pseudobulb diameter of the three-fork bletilla striata tissue culture seedling is larger than 6mm, arranging the domestication matrix in the fourth step in a plastic greenhouse with a sunshade net, and then transplanting the tissue culture seedling taken out of the bottle onto the domestication matrix according to the transplanting depth of 1.5cm and spraying gibberellin for domestication and culturing.
The further improvement is that: the sterilization treatment in the first step specifically comprises the steps of performing surface sterilization treatment on the capsule for 60s by using 75% alcohol, performing sterilization treatment for 12 minutes by using 0.1% mercury chloride, and finally cleaning to obtain the sterilization capsule.
The further improvement is that: in the second step, each liter of sowing culture medium comprises 1/2 MS, 1.0mg/L NAA, 5g/L agar, 30g/L white sugar and 65g/L mashed potato, and the pH value of the sowing culture medium is 5.8.
The further improvement is that: sowing according to the density of 13-18 bottles of seeds of a bletilla striata capsule during sowing, retaining condensed water during preparation of a sowing culture medium, slightly shaking a culture bottle after sowing seeds, and injecting sterile water into the culture bottle according to the amount of 1-2 mL/bottle when the condensed water during preparation of the culture medium is insufficient; immediately covering a culture bottle cap and marking after the sowing treatment is finished, placing the culture bottle in an inoculation chamber for 5 days of dark culture, then moving the culture bottle to the culture chamber for illumination culture under the condition of illumination intensity of 2000-2500 Lx at the temperature of 23-27 ℃ for 12 hours/day.
The further improvement is that: and in the third step, 80 mass percent and 600 times diluted mancozeb aqueous solution is used for carrying out soaking treatment on the roots of the seeding and culturing trigeminal bletilla striata seedlings for 3-5 min, and then the seedlings are transferred to a rooting and seedling strengthening culture medium.
The further improvement is that: in the third step, each liter of rooting and seedling strengthening culture medium comprises 1/2 MS, 0.5mg/L NAA, 4.6g/L agar, 30g/L white sugar and 100g/L mashed potato; the density of the tissue culture seedlings in a culture bottle is controlled to be 1-1.5 strain/cm during rooting and seedling strengthening culture 2 And culturing was performed in the culture chamber using the same light culture conditions as those used in sowing.
The further improvement is that: in the fourth step, the capsule shells are crushed and then are subjected to decomposition treatment with sheep manure synchronously, and then the decomposed capsule shells, coconut chaff, peat soil and decomposed sheep manure are mixed according to the mass ratio of 5% to 70% to 15% to 10% to prepare the domestication matrix.
The further improvement is that: and step five, uniformly spreading the acclimatized matrixes prepared by mixing according to the thickness of 12-15 cm when laying the acclimatized matrixes, and then compacting the soft acclimatized matrixes until the thickness of the acclimatized matrixes is compacted to the thickness of 8-10 cm.
The further improvement is that: spraying 1g/L gibberellin diluent to strengthen seedlings after illumination culture for three days in the step five, and watering by adopting a micro-spraying facility in the first month of the domestication culture to ensure that sprayed mist water permeates to the depth of 2cm of a domestication matrix; after domestication and culture for one month, the water spraying amount is increased according to the growth state of the trigeminal bletilla striata.
The beneficial effects of the application are as follows: according to the application, the problem of low bud growth rate after sowing due to incomplete disinfection is greatly reduced by carrying out deep disinfection by alcohol and mercury chloride, meanwhile, the root is soaked by using the mancozeb aqueous solution before rooting and transferring, so that the rooting survival rate is greatly increased, and the survival rate of the domesticated tissue culture seedlings of the trigeminal hyacinth bletilla is further improved by adding the decomposed capsule shells and sheep manure into the domestication matrix, the yield of the artificial planting of the trigeminal hyacinth bletilla is comprehensively increased, and the economic benefit of the planting of the trigeminal hyacinth bletilla is improved.
Drawings
FIG. 1 is a flow chart of a method according to embodiment 1 of the present application.
Detailed Description
The present application will be further described in detail with reference to the following examples, which are only for the purpose of illustrating the application and are not to be construed as limiting the scope of the application.
Example 1
According to the illustration in fig. 1, the embodiment provides a pure bletilla striata three-fork tissue culture domestication seedling raising method, which comprises the following steps:
firstly, deeply sterilizing, namely performing surface sterilization treatment on capsules by using 75% alcohol for 60 seconds under aseptic conditions, performing sterilization treatment by using 0.1% mercury chloride for 12 minutes, and repeatedly flushing the capsules by using sterile water for five times to obtain sterilized capsules;
wherein 0.1% of mercuric chloride is prepared by weighing 0.1 g of mercuric chloride, dissolving with a little alcohol, and adding distilled water to 100 ml.
Culturing after seed taking, namely sucking the cleaned disinfection capsules with sterile filter paper to dry surface water, then cutting the capsules with a sterile scalpel, uniformly shaking off internal seeds in a germination accelerator diluent by using sterile forceps, soaking for 5-8 hours, fishing out and draining, uniformly scattering on a sowing culture medium for sowing and culturing, wherein in order to reduce the difficulty of seed division, sowing is thin and not dense, and sowing is carried out according to the density of 13-18 bottles of seeds of the bletilla striata capsules; immediately covering a culture bottle cap and marking after the sowing treatment is finished, placing the culture bottle in an inoculation chamber for 5 days of dark culture, then moving the culture bottle to the culture chamber for illumination culture under the condition of illumination intensity of 2000-2500 Lx at the temperature of 23-27 ℃ for 12 hours/day;
wherein the seeding culture medium is prepared by mixing 1/2 MS, 1.0mg/L NAA, 5g/L agar, 30g/L white sugar and 65g/L mashed potato in a culture bottle, and the pH value of the seeding culture medium is 5.8; the condensed water is reserved when the sowing culture medium is prepared, and after the seeds are sown, the culture bottle is gently rocked to mix the seeds with the water, and the seeds are uniformly distributed on the surface of the culture medium; when the condensed water is insufficient in preparing the culture medium (namely, the condition that the surface of the culture medium is drier occurs), sterile water is injected into the culture flask according to the amount of 1-2 mL/flask so as to ensure that the seeds can fully absorb water and swell after being sown.
Step three, rooting and seedling strengthening culture, namely after seeding and culturing for two months, soaking the root of the trigeminal bletilla striata seedling which is induced to sprout in a mancozeb aqueous solution which is 80% of mass fraction and is diluted by 600 times, transferring the root of the trigeminal bletilla striata seedling to a rooting and seedling strengthening culture medium for strengthening culture of the tissue culture seedling, wherein the density of the tissue culture seedling in a culture bottle is controlled to be 1-1.5 strains/cm during rooting and seedling strengthening culture 2 Culturing in a culture room under the same illumination culture condition during sowing to promote the growth of pseudobulb of rhizoma Bletillae to a sufficient size for storageEnough nutrition is provided to ensure the survival rate of domestication of the tissue culture seedlings;
wherein the rooting and seedling strengthening culture medium is prepared by mixing 1/2 MS, 0.5mg/L NAA, 4.6g/L agar, 30g/L white sugar and 100g/L mashed potato in a culture bottle.
Step four, preparing an domestication matrix, namely crushing the capsule shells obtained by cutting and picking in the step two, and then uniformly mixing the capsule shells with coconut coir, peat soil and decomposed sheep manure to obtain the domestication matrix, wherein the domestication matrix is required to have better air permeability and water retention property;
wherein the capsule shells are crushed and then are decomposed together with sheep manure, then the decomposed capsule shells, coconut coir, peat soil and decomposed sheep manure are mixed according to the mass ratio of 5 percent to 70 percent to 15 percent to 10 percent to prepare an domestication matrix, and the decomposed capsule shells and the decomposed sheep manure are added to improve the domestication survival rate of the bletilla striata tissue culture seedlings.
Fifthly, domesticating and culturing, namely taking out the bottle when the pseudobulb diameter of the three-fork bletilla striata tissue culture seedling is larger than 6mm, arranging the domestication matrix in the fourth step in a plastic greenhouse with a sunshade net, and then transplanting the tissue culture seedling taken out of the bottle onto the domestication matrix according to the transplanting depth of 1.5cm and spraying gibberellin for domestication and culturing;
wherein, the domestication matrix is evenly spread according to the thickness of 12-15 cm when being laid, and then the soft domestication matrix is compacted until the thickness of the domestication matrix is compacted to 8-10 cm;
spraying 1g/L gibberellin diluent to strengthen seedlings after three days of illumination culture of domestication culture, and watering by adopting a micro-spraying facility in the first month of the domestication culture to ensure that sprayed atomized water permeates to the depth of 2cm of a domestication matrix, so that the air humidity can be kept and the matrix is not excessively wet; after domestication and culture for one month, the water spraying amount is increased according to the growth state of the trigeminal bletilla, but the excessive wetting of the matrix is avoided so as to prevent the pseudobulb of the trigeminal bletilla from rotting.
Example 2
The embodiment provides a planting contrast test of disinfection effect, germination rate and survival rate when using a pure bletilla striata three-fork tissue culture domestication seedling method to carry out seedling culture and traditional artificial domestication planting seedling culture, which specifically comprises the following steps:
firstly, two groups of capsules on the same strain of bletilla striata plant are obtained, the numbers A and B are respectively numbered, the bacterial strain content on single capsules of the two groups is detected and the detection result is counted before an experiment, then the capsules of the group A and the capsules of the group B are sterilized by adopting a single alcohol sterilization treatment method and the deep sterilization method according to the traditional manual domestication planting seedling culture method, and the bacterial strain content statistical detection result on single capsules is detected again after the sterilization treatment and the sterilization rate is calculated;
and then, continuously adopting the traditional domestication seedling method and the traditional domestication seedling method for the capsules in the group A and the capsules in the group B for seedling, and calculating the seedling germination rate and the survival rate of the capsules in the two groups by statistics during the period, wherein specific experimental data are shown in the following table.
Data sheet of sterilization effect, germination rate and survival rate of capsules in group A and group B
Compared with the traditional method, the method has the advantages that the sterilization rate, the germination rate and the survival rate are effectively improved, and the method means that the yield of artificial planting of the bletilla striata is effectively improved by domesticating and raising seedlings by the method, and the economic benefit of artificial planting is greatly improved.
The foregoing has shown and described the basic principles, principal features and advantages of the application. It will be understood by those skilled in the art that the present application is not limited to the embodiments described above, and that the above embodiments and descriptions are merely illustrative of the principles of the present application, and various changes and modifications may be made without departing from the spirit and scope of the application, which is defined in the appended claims. The scope of the application is defined by the appended claims and equivalents thereof.
Claims (9)
1. The method for cultivating and domesticating pure bletilla striata three-fork tissue culture seedlings is characterized by comprising the following steps of:
the method comprises the steps of firstly, deeply sterilizing, sterilizing the mature and uncracked capsule of the bletilla striata by using alcohol and 0.1% mercury chloride under a sterile condition, and repeatedly flushing the capsule with sterile water for five times to obtain a sterilized capsule;
culturing after seed taking, namely sucking the surface water of the cleaned disinfection capsules by using sterile filter paper, cutting the capsules by using a disinfection scalpel, uniformly shaking off the internal seeds by using a sterile forceps, soaking the seeds in a germination accelerator diluent for 5-8 hours, fishing out the seeds, draining the seeds, and uniformly scattering the seeds on a sowing culture medium for sowing culture;
step three, rooting and seedling strengthening culture, namely, after sowing and culturing for two months, soaking the trigeminal bletilla striata seedlings which induce sprouting in a mancozeb aqueous solution, transferring the soaked trigeminal bletilla striata seedlings to a rooting and seedling strengthening culture medium, and performing seedling strengthening culture on the tissue culture seedlings;
step four, preparing an domestication matrix, namely crushing the capsule shells obtained by cutting and seed taking in the step two, and then uniformly mixing the capsule shells with coconut coir, peat soil and decomposed sheep manure to obtain the domestication matrix;
fifthly, domesticating and culturing, namely taking out the bottle when the pseudobulb diameter of the tissue culture seedling of the trigeminal bletilla is larger than 6mm, arranging the domestication matrix in the fourth step in a plastic greenhouse with a sunshade net, then transplanting the tissue culture seedling taking out the bottle onto the domestication matrix according to the transplanting depth of 1.5cm, and spraying gibberellin for domestication and culturing, so that the trigeminal bletilla is obtained.
2. The method for cultivating and domesticating pure bletilla striata three-fork tissue culture seedlings according to claim 1, wherein the method comprises the following steps: the sterilization treatment in the first step specifically comprises the steps of performing surface sterilization treatment on the capsule for 60s by using 75% alcohol, performing sterilization treatment for 12 minutes by using 0.1% mercury chloride, and finally cleaning to obtain the sterilization capsule.
3. The method for cultivating and domesticating pure bletilla striata three-fork tissue culture seedlings according to claim 1, wherein the method comprises the following steps: in the second step, each liter of sowing culture medium comprises 1/2 MS, 1.0mg/L NAA, 5g/L agar, 30g/L white sugar and 65g/L mashed potato, and the pH value of the sowing culture medium is 5.8.
4. The method for cultivating and domesticating pure bletilla striata three-fork tissue culture seedlings according to claim 1, wherein the method comprises the following steps: sowing according to the density of 13-18 bottles of seeds of a bletilla striata capsule during sowing, retaining condensed water during preparation of a sowing culture medium, slightly shaking a culture bottle after sowing seeds, and injecting sterile water into the culture bottle according to the amount of 1-2 mL/bottle when the condensed water during preparation of the culture medium is insufficient; immediately covering a culture bottle cap and marking after the sowing treatment is finished, placing the culture bottle in an inoculation chamber for 5 days of dark culture, then moving the culture bottle to the culture chamber for illumination culture under the condition of illumination intensity of 2000-2500 Lx at the temperature of 23-27 ℃ for 12 hours/day.
5. The method for cultivating and domesticating pure bletilla striata three-fork tissue culture seedlings according to claim 1, wherein the method comprises the following steps: and in the third step, 80 mass percent and 600 times diluted mancozeb aqueous solution is used for carrying out soaking treatment on the roots of the seeding and culturing trigeminal bletilla striata seedlings for 3-5 min, and then the seedlings are transferred to a rooting and seedling strengthening culture medium.
6. The method for cultivating and domesticating pure bletilla striata three-fork tissue culture seedlings according to claim 1, wherein the method comprises the following steps: in the third step, each liter of rooting and seedling strengthening culture medium comprises 1/2 MS, 0.5mg/L NAA, 4.6g/L agar, 30g/L white sugar and 100g/L mashed potato; the density of the tissue culture seedlings in a culture bottle is controlled to be 1-1.5 strain/cm during rooting and seedling strengthening culture 2 And culturing was performed in the culture chamber using the same light culture conditions as those used in sowing.
7. The method for cultivating and domesticating pure bletilla striata three-fork tissue culture seedlings according to claim 1, wherein the method comprises the following steps: in the fourth step, the capsule shells are crushed and then are subjected to decomposition treatment with sheep manure synchronously, and then the decomposed capsule shells, coconut chaff, peat soil and decomposed sheep manure are mixed according to the mass ratio of 5% to 70% to 15% to 10% to prepare the domestication matrix.
8. The method for cultivating and domesticating pure bletilla striata three-fork tissue culture seedlings according to claim 1, wherein the method comprises the following steps: and step five, uniformly spreading the acclimatized matrixes prepared by mixing according to the thickness of 12-15 cm when laying the acclimatized matrixes, and then compacting the soft acclimatized matrixes until the thickness of the acclimatized matrixes is compacted to the thickness of 8-10 cm.
9. The method for cultivating and domesticating pure bletilla striata three-fork tissue culture seedlings according to claim 1, wherein the method comprises the following steps: spraying 1g/L gibberellin diluent to strengthen seedlings after illumination culture for three days in the step five, and watering by adopting a micro-spraying facility in the first month of the domestication culture to ensure that sprayed mist water permeates to the depth of 2cm of a domestication matrix; after domestication and culture for one month, the water spraying amount is increased according to the growth state of the trigeminal bletilla striata.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310748215.0A CN116649216A (en) | 2023-06-25 | 2023-06-25 | Pure bletilla striata three-fork tissue culture domesticated seedling raising method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310748215.0A CN116649216A (en) | 2023-06-25 | 2023-06-25 | Pure bletilla striata three-fork tissue culture domesticated seedling raising method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116649216A true CN116649216A (en) | 2023-08-29 |
Family
ID=87724106
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310748215.0A Pending CN116649216A (en) | 2023-06-25 | 2023-06-25 | Pure bletilla striata three-fork tissue culture domesticated seedling raising method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116649216A (en) |
Citations (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104145816A (en) * | 2014-07-29 | 2014-11-19 | 郎溪县紫兰药材种植专业合作社 | Bletilla striata tissue-culture seedling raising method |
| CN104719152A (en) * | 2015-02-16 | 2015-06-24 | 贵州省农作物品种资源研究所 | Rhizoma bletillae industrialized seedling method |
| CN104737911A (en) * | 2015-03-30 | 2015-07-01 | 云南中医学院 | Quick cultivation method for rhizoma bletillae tissue culture seedlings |
| CN104782485A (en) * | 2015-04-08 | 2015-07-22 | 安徽春之蔚农业科技有限公司 | Method for rapid tissue propagation and breeding seedling of bletilla striata seeds |
| CN104904597A (en) * | 2015-05-28 | 2015-09-16 | 云南省农业科学院农业环境资源研究所 | Bletilla striata rapid propagation and sunshine seedling strengthening method |
| CN105103928A (en) * | 2015-09-29 | 2015-12-02 | 江苏农林职业技术学院 | Method for acclimatizing and transplanting bletilla striata tissue culture seedlings |
| CN105145352A (en) * | 2015-08-03 | 2015-12-16 | 河南科技大学 | Efficient tissue culture and rapid propagation technology for seedlings of bletilla striata |
| CN106305055A (en) * | 2016-08-18 | 2017-01-11 | 安徽省彩枫农林科技有限公司 | Domestication method for rhizoma bletillae seedlings |
| CN106417011A (en) * | 2016-08-30 | 2017-02-22 | 云南金碧制药有限公司 | Wild bletilla striata tissue culture rapid propagation method |
| CN107047310A (en) * | 2017-05-10 | 2017-08-18 | 文山学院 | A kind of cultural method of bletilla striata seeds culture seedling |
| CN108541589A (en) * | 2018-03-22 | 2018-09-18 | 恩施土家族苗族自治州农业科学院 | A kind of rapid propagation method of the bletilla striata |
| CN108651275A (en) * | 2017-03-27 | 2018-10-16 | 云南省德宏热带农业科学研究所 | A kind of method of quickly breeding bletilla striata seedling under natural light |
| CN109169275A (en) * | 2018-09-13 | 2019-01-11 | 甘肃源宜生物科技有限公司 | A kind of pale reddish brown trident bletilla striata tissue-culturing rapid propagation culture medium and method |
| CN109964815A (en) * | 2019-03-26 | 2019-07-05 | 成都大学 | A kind of bletilla striata aseptic seedling rapid induction Multiple Buds and fast numerous method of taking root |
| CN110547194A (en) * | 2019-09-12 | 2019-12-10 | 浙江省东阳玉米研究所 | Tissue culture and rapid propagation method for seeds of bletilla striata |
| CN112889672A (en) * | 2021-03-19 | 2021-06-04 | 冯耀文 | Cultivation method for high-quality and high-yield bletilla striata seedlings |
-
2023
- 2023-06-25 CN CN202310748215.0A patent/CN116649216A/en active Pending
Patent Citations (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104145816A (en) * | 2014-07-29 | 2014-11-19 | 郎溪县紫兰药材种植专业合作社 | Bletilla striata tissue-culture seedling raising method |
| CN104719152A (en) * | 2015-02-16 | 2015-06-24 | 贵州省农作物品种资源研究所 | Rhizoma bletillae industrialized seedling method |
| CN104737911A (en) * | 2015-03-30 | 2015-07-01 | 云南中医学院 | Quick cultivation method for rhizoma bletillae tissue culture seedlings |
| CN104782485A (en) * | 2015-04-08 | 2015-07-22 | 安徽春之蔚农业科技有限公司 | Method for rapid tissue propagation and breeding seedling of bletilla striata seeds |
| CN104904597A (en) * | 2015-05-28 | 2015-09-16 | 云南省农业科学院农业环境资源研究所 | Bletilla striata rapid propagation and sunshine seedling strengthening method |
| CN105145352A (en) * | 2015-08-03 | 2015-12-16 | 河南科技大学 | Efficient tissue culture and rapid propagation technology for seedlings of bletilla striata |
| CN105103928A (en) * | 2015-09-29 | 2015-12-02 | 江苏农林职业技术学院 | Method for acclimatizing and transplanting bletilla striata tissue culture seedlings |
| CN106305055A (en) * | 2016-08-18 | 2017-01-11 | 安徽省彩枫农林科技有限公司 | Domestication method for rhizoma bletillae seedlings |
| CN106417011A (en) * | 2016-08-30 | 2017-02-22 | 云南金碧制药有限公司 | Wild bletilla striata tissue culture rapid propagation method |
| CN108651275A (en) * | 2017-03-27 | 2018-10-16 | 云南省德宏热带农业科学研究所 | A kind of method of quickly breeding bletilla striata seedling under natural light |
| CN107047310A (en) * | 2017-05-10 | 2017-08-18 | 文山学院 | A kind of cultural method of bletilla striata seeds culture seedling |
| CN108541589A (en) * | 2018-03-22 | 2018-09-18 | 恩施土家族苗族自治州农业科学院 | A kind of rapid propagation method of the bletilla striata |
| CN109169275A (en) * | 2018-09-13 | 2019-01-11 | 甘肃源宜生物科技有限公司 | A kind of pale reddish brown trident bletilla striata tissue-culturing rapid propagation culture medium and method |
| CN109964815A (en) * | 2019-03-26 | 2019-07-05 | 成都大学 | A kind of bletilla striata aseptic seedling rapid induction Multiple Buds and fast numerous method of taking root |
| CN110547194A (en) * | 2019-09-12 | 2019-12-10 | 浙江省东阳玉米研究所 | Tissue culture and rapid propagation method for seeds of bletilla striata |
| CN112889672A (en) * | 2021-03-19 | 2021-06-04 | 冯耀文 | Cultivation method for high-quality and high-yield bletilla striata seedlings |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103141260B (en) | Method for ecologically interplanting sarcandra glabra under phyllostachys edulis forest | |
| CN103918426B (en) | A kind of high yield of loofah implantation methods | |
| CN104170656B (en) | Grafting method for thin-skin muskmelons | |
| CN103444530A (en) | Technology for field return-to-nature ex-situ plantation of wild anoectochilus roxburghii (Wall.) Lindl in Fujian | |
| CN106577281A (en) | High-seedling-rate culture method for tissue culture of Polygala fallax stems | |
| CN106359087A (en) | Tissue culture quick-breeding seedling raising method for radix asparagi | |
| CN108934679B (en) | Root-cutting seedling raising and seedling tube raising method for aralia elata seem | |
| CN113170707B (en) | Efficient seedling growing method for mesona blume virus-free seedlings and application of efficient seedling growing method | |
| CN1973605B (en) | Artificial propagation process of Dracaena cochinchinensis | |
| CN105875412B (en) | The implantation methods of organic selenium-rich sugariness gynostemma pentaphylla | |
| CN111657068A (en) | Method for breeding epimedium seedlings | |
| CN104604680B (en) | Can promote that bletilla striata seeds sprouts the culture medium with growth of seedling and formula thereof and preparation method | |
| CN114600891A (en) | Composition and method for seed germination | |
| CN110915361B (en) | Angelica keiskei germination accelerating method | |
| CN108029555A (en) | A kind of bletilla striata method for culturing seedlings | |
| CN107517838A (en) | A kind of dendrobium candidum gemmule method for culturing seedlings | |
| CN116649216A (en) | Pure bletilla striata three-fork tissue culture domesticated seedling raising method | |
| CN111480579A (en) | Tissue culture and rapid propagation method for immature embryos of epimedium dauricum | |
| CN111557241A (en) | Rapid propagation method and application of bighead atractylodes rhizome seedlings | |
| CN111296194A (en) | Disease control method based on strawberry seedling planting process | |
| CN1631112A (en) | Tissue culture breeding and plating technique of 'shejiegu' | |
| CN108207464A (en) | A kind of bletilla striata high-yield planting method under camellia oleifera lam | |
| CN108575521A (en) | A kind of method of Ling Yaoxing and red bayberry interplanting | |
| CN115715525B (en) | Tissue culture and rapid propagation method of calyx parviflora | |
| CN116584383B (en) | Establishment method of eucalyptus citriodora tissue culture seedling system |
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 |