CN110692522A - Tissue culture seedling method for Prunus cistana - Google Patents
Tissue culture seedling method for Prunus cistana Download PDFInfo
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- CN110692522A CN110692522A CN201911192920.7A CN201911192920A CN110692522A CN 110692522 A CN110692522 A CN 110692522A CN 201911192920 A CN201911192920 A CN 201911192920A CN 110692522 A CN110692522 A CN 110692522A
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- 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
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- 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
Abstract
The invention discloses a tissue culture seedling method of Prunus cistana, relates to plant propagation, and belongs to seedling production technology in forestry biotechnology. The method is characterized in that: fumigating and sterilizing an intelligent seedling raising greenhouse, selecting a Prunus davidiana stem section, cutting the Prunus davidiana stem section into 2-3 cm stem sections with top buds after surface sterilization, placing the stem sections on a reciprocating shaking seedbed, and culturing by using an adventitious bud culture medium added with 6-BA (1.0 mg/l) and NAA (0.05 mg/l); when the plant grows to 2.5-3.5 cm, transferring to the other half of reciprocating shaking seedbed, inducing rooting by using rooting culture medium added with IBA (1.0 mg/l) and IAA (1.0 mg/l), after 35d, hardening the seedling for 5d, and transferring to a nutrition pot for propagation. The invention combines the traditional seedling raising method and the tissue culture seedling raising method together, avoids the defects that the death of a culture is easy to cause by the oxygen deficiency of a liquid culture medium and the growth of a solid culture medium is slow by shaking the shallow layer of the seedbed in a reciprocating mode for shake culture, achieves the aim of large-scale rapid propagation, and has simple process and easy popularization.
Description
Technical Field
The invention relates to a rapid seedling propagation production technology in forestry biotechnology, in particular to a tissue culture seedling method of Prunus davidiana.
Background
Prunus cistana (a. juss.)Prunus× cistena N.E.Hansen ex Koehne) The small deciduous tree or shrub belongs to Rosaceae, Prunus, the plant height is 1.5-2.6 m, the crown width is 1.2-2.9 m, the young branch is purple brown, usually has no hair, the old branch has skin holes, the small deciduous tree or shrub is distributed on the whole branch, single leaves are intergrown, the leaves are long oval or oval and long elliptical, the length is 4-8 cm, the tip is gradually sharp, the base of the leaf is wide and wedge-shaped, the leaf edge has uneven thin and blunt teeth, the leaf surface is red or purple, and the top of the new leaf is bright purplish red. The flower is single-born, small, light pink, 5 petals, slight fragrance, most stamens, single pistil and the flowering period of 4-5 months.
The Prunus davidiana var is a hybrid of Prunus davidiana var and Prunus davidiana var, and is cultivated all over the country. The Prunus cistana nakai has strong adaptability, cold resistance and disease resistance, and can grow well on fertile sandy soil and mild clay. The purple-leaf dwarf cherry has strong sprouting ability, luxuriant branches and leaves, and purplish red branches and leaves, is a famous leaf-viewing tree species in the world, is a material selected for manufacturing color balls, color bands, hedgerows and the like due to strong pruning resistance and more gorgeous leaf color after pruning, is an excellent color leaf configured tree species for urban landscaping, and has great market development potential.
The short cherry with purple leaves is generally propagated by grafting and cutting, and is cut-grafted in spring and autumn or bud-grafted in summer and autumn, so that mass propagation is inconvenient. And because the seedlings of the short cherries with purple leaves are slow, the seedlings can be applied to engineering generally for at least three years, the time of four to five is needed for the true growth, and obviously, the traditional grafting and cutting propagation modes can not meet the requirements of large-scale landscaping engineering construction. Therefore, how to accelerate the propagation speed of the short cerasus serrulata koch is very important, the invention manually controls the conditions of temperature, illumination, humidity, nutrition, hormone and the like in a specific place and environment by a tissue culture method, automatically controls the production, ensures that the branches of the short cerasus serrulata koch are rapidly propagated in large quantity, selects the stem section with terminal buds of the short cerasus serrulata koch as a material, can obtain more than 50 seedlings from 1 branch in one year, achieves the purpose of large-scale rapid asexual propagation, can effectively save manpower and material resources, and has high survival rate.
Disclosure of Invention
The invention aims to provide a tissue culture seedling raising method of Prunus cistana, which can accelerate the seedling breeding speed, shorten the seedling raising period, reduce the risk of seedling planting industry, obviously improve the survival rate of Prunus cistana seedlings, and has simple and easy operation, thereby providing guarantee for rapid and high-quality seedling culture and breeding.
In order to achieve the purpose, the tissue culture seedling method of the Prunus cistana is characterized in that: selecting stem ends with terminal buds of Prunus davidiana as propagation materials, sterilizing the surfaces of the propagation materials, cutting the propagation materials, taking stem sections with terminal buds cut into 2-3 cm, taking a liquid adventitious bud culture medium as a basic culture medium, performing shallow shaking culture by a reciprocating shaking table, adding growth regulators 6-BA (1.0 mg/l) and NAA (0.05mg/l) for culture, and inducing adventitious bud differentiation; transferring the plant to a liquid rooting culture medium when the plant grows to 2.5-3.5 cm, carrying out shake culture on the plant through a reciprocating type shaking table in a shallow layer, adding growth regulators IBA (1.0 mg/l) and IAA (1.0 mg/l) to induce rooting, inoculating for 35d, hardening the rooted seedling for 5d, cutting off larger leaves, and transferring the seedling to a nutrition pot for culture and propagation.
The adventitious bud culture medium: the following substances in mg/L: 660 parts of ammonium nitrate, 760 parts of potassium nitrate, 176 parts of calcium chloride dihydrate, 148 parts of magnesium sulfate heptahydrate, 68 parts of monopotassium phosphate, 1.66 parts of potassium iodide, 12.4 parts of boric acid, 44.6 parts of manganese sulfate tetrahydrate, 17.2 parts of zinc sulfate heptahydrate, 0.5 part of sodium molybdate dihydrate, 0.05 part of copper sulfate pentahydrate, 0.05 part of cobalt chloride hexahydrate and 55.6 parts of ferric sulfate heptahydrate; 74.6 parts of disodium ethylene diamine tetraacetate, 200 parts of inositol, 1 part of nicotinic acid, 1 part of pyridoxine hydrochloride, 1 part of thiamine hydrochloride, 4 parts of glycine, 3500 parts of cane sugar and the balance of water, adjusting the pH value of a culture medium to 5.7-5.8 by using sodium hydroxide, sterilizing at high temperature and sealing for later use.
The rooting culture medium comprises: the following substances in mg/L: 330 parts of ammonium nitrate, 380 parts of potassium nitrate, 88 parts of calcium chloride dihydrate, 74 parts of magnesium sulfate heptahydrate, 34 parts of monopotassium phosphate, 0.83 part of potassium iodide, 6.2 parts of boric acid, 22.3 parts of manganese sulfate tetrahydrate, 8.6 parts of zinc sulfate heptahydrate, 0.25 part of sodium molybdate dihydrate, 0.025 part of copper sulfate pentahydrate, 0.025 part of cobalt chloride hexahydrate and 27.8 parts of ferric sulfate heptahydrate; 37.3 parts of disodium ethylene diamine tetraacetate, 100 parts of inositol, 0.5 part of nicotinic acid, 0.5 part of pyridoxine hydrochloride, 0.5 part of thiamine hydrochloride, 2 parts of glycine, 3500 parts of cane sugar and the balance of water, adjusting the pH value of the culture medium to 5.7-5.8 by using sodium hydroxide, sterilizing at high temperature and sealing for later use.
The invention relates to a tissue culture seedling method of Prunus cistana, which is realized by the following steps:
firstly, selection and requirements of an intelligent seedling raising greenhouse:
the intelligent seedling raising greenhouse is required to be closed and clean, the temperature, the humidity and the illumination are controllable, the room temperature is uniform and consistent, the heat insulation performance is realized, and an ultraviolet lamp, an air sterilizer or other sterilization equipment is arranged, so that the sterilization can be carried out at any time;
secondly, indoor fumigation and disinfection:
thoroughly fumigating and disinfecting an intelligent seedling raising greenhouse before inoculation, wherein the method comprises the steps of driving atomization by using the amount of 10-15 ml of hydrogen peroxide and 6-7.5 g of potassium permanganate per cubic meter, sealing a sterilization space for 36-72 hours, and exhausting air for 1 hour; opening an ultraviolet lamp and an air sterilizer for sterilization before inoculating for 5h, reducing the exchange between the inoculating room and the outside air and the air entering the room after inoculating, and filtering and cleaning the air;
thirdly, setting of the reciprocating type rocking seedbed:
the reciprocating type rocking seedbed comprises the following specific setting steps:
(1) arranging the reciprocating type shaking tables in a straight line, placing the disposable sealed clean porous seedling raising tray on the shaking tables to be used as a culture container, and locking the culture container by using a mold clamp to form the reciprocating type shaking seedbed, wherein the shaking amplitude of the shaking tables is set to be 20mm, and the shaking frequency is set to be 60 rpm;
(2) reserving a half of the culture container in the step (1), tearing a seedling raising tray sealing film, and filling a liquid adventitious bud culture medium prepared in advance for later use; after about 9 days, according to the culture requirement, tearing off the other half of the seedling raising tray sealing film, and filling the seedling raising tray sealing film into a pre-prepared rooting culture medium for later use;
fourthly, selecting in vitro propagation materials:
selecting plants with good genetic stability and retaining the excellent characters of the Prunus cerasifera; in sunny days, taking the stem end with strong regeneration capacity and terminal bud as an in vitro propagation material, and storing the stem end in a cool and humid place for later use;
fifthly, processing the explants:
washing the selected in-vitro propagation materials with tap water, shearing the materials into 2-3 cm in size, adding washing powder for secondary deep washing if necessary, then putting the materials into 70% alcohol for disinfection for 10-25 s, then soaking the materials in sodium hypochlorite for 18min, and finally putting the materials into a sterile container for inoculation;
sixthly, differentiation of adventitious buds:
inoculating 2-3 cm stem segments with terminal buds to an adventitious bud culture medium, opening a reciprocating type shaking table for shallow shake culture, adding growth regulators 6-BA (1.0 mg/l) and NAA (0.05mg/l) to induce adventitious bud differentiation, culturing at 25 +/-3 ℃, wherein the illumination intensity is 1000lx to 6000lx, illuminating for 16h and 8h in darkness for one day, the illumination intensity is gradually enhanced along with organ differentiation, the humidity is 70-80% of relative humidity, and culturing for about 9d to ensure that green bud spots appear on the edges of explants;
seventhly, rooting induction:
transferring the plant to a rooting culture medium when the plant grows to 2.5-3.5 cm, opening a reciprocating type shaking table to perform shallow shaking culture, adding growth regulators IBA (1.0 mg/l) and IAA (1.0 mg/l) to induce rooting, wherein the culture temperature is 25 +/-3 ℃, the illumination intensity is 1000lx to 6000lx, the plant is illuminated for 16h and 8h in darkness in one day, the illumination intensity is continuously enhanced along with the differentiation of organs and the growth of seedlings, the humidity is 70-80% relative humidity, and culturing for 35d to obtain seedlings;
eighthly, transplanting the tissue culture seedlings:
hardening the rooted seedlings for 5 days, cutting off large leaves, transplanting the seedlings into a nutrition pot with dried and disinfected culture soil for propagation culture, culturing the domesticated tissue culture seedlings in a greenhouse for a period of time, and transplanting the domesticated tissue culture seedlings to open field for cultivation as required.
The tissue culture seedling method of the Prunus cistana, disclosed by the invention, has the beneficial effects that:
(1) the stem section with the terminal bud of the Prunus cerasifera is selected as a propagation material, and shallow layer shaking culture is realized through a reciprocating type shaking table, and the method of regular immersion effectively avoids the defects that the death of a culture is easily caused by the lack of oxygen in a liquid culture medium and the growth of a solid culture medium is slow;
(2) the traditional seedling culture method and the tissue culture seedling culture method are organically combined together, the materials are easy to obtain, the operation is convenient, the simplicity and the high efficiency are realized, the culture condition is controllable, the propagation can be carried out all the year round, and compared with the grafting and cutting propagation modes, a series of complicated labor such as weeding, fertilizing, watering, pest control and the like is saved;
(3) the method has the advantages of small floor area, short propagation period, high propagation coefficient, adventitious bud induction rate of 85.6 percent and rooting rate of 81.2 percent, can obtain more than 50 seedlings from 1 branch in one year, is favorable for highly intensive factory production, ensures that the Prunus cerasifera branches are rapidly propagated in large quantities, and has high survival rate.
Detailed Description
The following examples further illustrate the invention.
Example 1
Firstly, selection and requirements of an intelligent seedling raising greenhouse:
the intelligent seedling raising greenhouse is required to be closed and clean, the temperature, the humidity and the illumination are controllable, the room temperature is uniform and consistent, the heat insulation performance is realized, and an ultraviolet lamp, an air sterilizer or other sterilization equipment is arranged, so that the sterilization can be carried out at any time;
secondly, indoor fumigation and disinfection:
thoroughly fumigating and disinfecting an intelligent seedling raising greenhouse before inoculation, wherein the method comprises the steps of driving atomization by using 10-15 ml of hydrogen peroxide and 6-7.5 g of potassium permanganate per cubic meter, sealing a sterilization space for 36-72 hours, and exhausting air for 1 hour; opening an ultraviolet lamp and an air sterilizer for sterilization before inoculating for 5 h; after inoculation is started, exchange with outside air is reduced as much as possible, and air entering a room is filtered and cleaned;
thirdly, setting of the reciprocating type rocking seedbed:
the reciprocating type rocking seedbed comprises the following specific setting steps:
(1) arranging the reciprocating type shaking tables in a straight line, placing the disposable sealed clean porous seedling raising tray on the shaking tables to be used as a culture container, and locking the culture container by using a mold clamp to form the reciprocating type shaking seedbed, wherein the shaking amplitude of the shaking tables is set to be 20mm, and the shaking frequency is set to be 60 rpm;
(2) reserving a half of the culture container in the step (1), tearing a seedling raising tray sealing film, and filling a liquid adventitious bud culture medium prepared in advance for later use; after about 9 days, according to the culture requirement, tearing off the other half of the seedling raising tray sealing film, and filling the seedling raising tray sealing film into a pre-prepared rooting culture medium for later use; adventitious bud culture medium: the following substances in mg/L: 660 parts of ammonium nitrate, 760 parts of potassium nitrate, 176 parts of calcium chloride dihydrate, 148 parts of magnesium sulfate heptahydrate, 68 parts of monopotassium phosphate, 1.66 parts of potassium iodide, 12.4 parts of boric acid, 44.6 parts of manganese sulfate tetrahydrate, 17.2 parts of zinc sulfate heptahydrate, 0.5 part of sodium molybdate dihydrate, 0.05 part of copper sulfate pentahydrate, 0.05 part of cobalt chloride hexahydrate and 55.6 parts of ferric sulfate heptahydrate; 74.6 parts of disodium ethylene diamine tetraacetate, 200 parts of inositol, 1 part of nicotinic acid, 1 part of pyridoxine hydrochloride, 1 part of thiamine hydrochloride, 4 parts of glycine, 3500 parts of cane sugar and the balance of water, wherein the PH value is 5.7-5.8; rooting culture medium: the following substances in mg/L: 330 parts of ammonium nitrate, 380 parts of potassium nitrate, 88 parts of calcium chloride dihydrate, 74 parts of magnesium sulfate heptahydrate, 34 parts of monopotassium phosphate, 0.83 part of potassium iodide, 6.2 parts of boric acid, 22.3 parts of manganese sulfate tetrahydrate, 8.6 parts of zinc sulfate heptahydrate, 0.25 part of sodium molybdate dihydrate, 0.025 part of copper sulfate pentahydrate, 0.025 part of cobalt chloride hexahydrate and 27.8 parts of ferric sulfate heptahydrate; 37.3 parts of disodium ethylene diamine tetraacetate, 100 parts of inositol, 0.5 part of nicotinic acid, 0.5 part of pyridoxine hydrochloride, 0.5 part of thiamine hydrochloride, 2 parts of glycine, 3500 parts of cane sugar and the balance of water, wherein the pH value is 5.7-5.8;
fourthly, selecting in vitro propagation materials:
selecting plants with good genetic stability and retaining the excellent characters of the Prunus cerasifera; taking the stem end with apical bud with strong regeneration capability on the robust mother branch as an in vitro propagation material, and the material with wound or plant diseases and insect pests can not be taken; in addition, the material taking time is selected in a sunny day, the material is prevented from being taken in a rainy day or when dew is not dry, and the sheared propagation materials are stored in a cool and humid place for later use;
fifthly, processing the explants:
the specific process of the explant treatment comprises the following steps:
(1) removing unused parts of the collected propagation materials, and cleaning required parts; then, shearing the material into 2-3 cm;
(2) washing the propagation material subjected to primary cleaning for 3-5 min by using tap water, adding washing powder to thoroughly clean lipidated substances on the surface of the propagation material if necessary, and washing for 3-5 min by using the tap water;
(3) putting the propagation material subjected to deep cleaning into 70% alcohol for disinfection for 10-25 s, then soaking the propagation material in sodium hypochlorite for 18min, and finally putting the propagation material into a sterile container for inoculation; it should be noted that the alcohol infiltration time cannot be too long because the alcohol penetration is strong and is easy to kill cells;
sixthly, differentiation of adventitious buds:
taking stem segments with terminal buds cut into 2-3 cm, inoculating the stem segments to an adventitious bud culture medium, opening a reciprocating type shaking table for shallow shaking culture, adding growth regulators 6-BA (1.0 mg/l) and NAA (0.05mg/l) to induce the adventitious bud to differentiate, wherein the culture temperature is 25 +/-3 ℃, the illumination intensity is 1000lx to 6000lx, the illumination is 16h and 8h in darkness every day, the illumination intensity is gradually enhanced along with the differentiation of organs, the humidity is 70-80% relative humidity, and green bud spots appear on the edges of explants after about 9d of culture;
seventhly, rooting induction:
transferring the plant to a rooting culture medium when the plant grows to 2.5-3.5 cm, opening a reciprocating type shaking table to perform shallow shaking culture, adding growth regulators IBA (1.0 mg/l) and IAA (1.0 mg/l) to induce rooting, wherein the culture temperature is 25 +/-3 ℃, the illumination intensity is 1000lx to 6000lx, the plant is illuminated for 16h and 8h in darkness in one day, the illumination intensity is continuously enhanced along with the differentiation of organs and the growth of seedlings, the humidity is 70-80% relative humidity, and culturing for 35d to obtain seedlings;
eighthly, transplanting the tissue culture seedlings:
hardening the rooted seedlings for 5 days, cutting off large leaves, transplanting the seedlings into a nutrition pot with dried and disinfected culture soil for propagation culture, culturing the domesticated tissue culture seedlings in a greenhouse for a period of time, and transplanting the domesticated tissue culture seedlings to open field for cultivation as required.
The above-mentioned embodiments only express a certain implementation way of the present invention, and do not limit the patent scope, and it is obvious for those skilled in the art that on the basis of the above description, several variations and modifications can be made to the present invention, which all belong to the protection scope of the present invention; therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (6)
1. A tissue culture seedling method of Prunus cistana, which is characterized in that: thoroughly fumigating and sterilizing an intelligent seedling raising greenhouse, selecting stem sections with terminal buds of Prunus davidiana as in-vitro propagation materials, sterilizing the surfaces of the stem sections, cutting the stem sections with the terminal buds of 2-3 cm, placing the stem sections on a sterilized and sterilized reciprocating shaking seedbed, performing shallow layer shaking culture by using a liquid adventitious bud culture medium, adding a growth regulator 6-BA (1.0 mg/l) and NAA (0.05mg/l) for culture, and inducing adventitious bud differentiation; when the plant grows to 2.5-3.5 cm, transferring to a reciprocating shaking seedbed, carrying out shallow shaking culture by using a liquid rooting culture medium, adding growth regulators IBA (1.0 mg/l) and IAA (1.0 mg/l) to induce rooting, inoculating for 35d, hardening the rooted seedlings for 5d, cutting off larger leaves, and transferring to a nutrition pot for culture and propagation;
the adventitious bud culture medium: the following substances in mg/L: potassium nitrate 760, ammonium nitrate 660, calcium chloride dihydrate 176, magnesium sulfate heptahydrate 148, monopotassium phosphate 68, manganese sulfate tetrahydrate 44.6, zinc sulfate heptahydrate 17.2, boric acid 12.4, potassium iodide 1.66, cobalt chloride hexahydrate 0.05, copper sulfate pentahydrate 0.05, sodium molybdate dihydrate 0.5, disodium ethylene diamine tetraacetate 74.6, ferric sulfate heptahydrate 55.6, inositol 200, glycine 4, nicotinic acid 1, thiamine hydrochloride 1, pyridoxine hydrochloride 1, sucrose 3500 and the balance of water, wherein the pH is 5.7-5.8;
the rooting culture medium comprises: the following substances in mg/L: potassium nitrate 380, ammonium nitrate 330, calcium chloride dihydrate 88, magnesium sulfate heptahydrate 74, monopotassium phosphate 34, manganese sulfate tetrahydrate 22.3, zinc sulfate heptahydrate 8.6, boric acid 6.2, potassium iodide 0.83, cobalt chloride hexahydrate 0.025, copper sulfate pentahydrate 0.025, sodium molybdate dihydrate 0.25, disodium ethylene diamine tetraacetate 37.3, ferric sulfate heptahydrate 27.8, inositol 100, glycine 2, nicotinic acid 0.5, thiamine hydrochloride 0.5, pyridoxine hydrochloride 0.5, sucrose 3500 and the balance of water, wherein the pH value is 5.7-5.8.
2. The tissue culture seedling method of the Prunus cerasifera as claimed in claim 1, which is characterized in that: thoroughly fumigating and disinfecting an intelligent seedling raising greenhouse before inoculation, wherein the method comprises the steps of driving atomization by using the amount of 10-15 ml of hydrogen peroxide and 6-7.5 g of potassium permanganate per cubic meter, sealing a sterilization space for 36-72 hours, and exhausting air for 1 hour; and (3) opening an ultraviolet lamp and an air sterilizer for sterilization before inoculating for 5h, and filtering and cleaning the air entering the room.
3. The tissue culture seedling method of the Prunus cerasifera as claimed in claim 1, which is characterized in that: and (3) sterilizing the in-vitro propagation material, namely washing the stem end with the terminal bud with tap water, putting the stem end into 70% alcohol for disinfection for 15-25 s, and soaking the stem end with sodium hypochlorite for 18 min.
4. The tissue culture seedling method of the Prunus cerasifera as claimed in claim 1, which is characterized in that: inducing adventitious bud differentiation, selecting 2-3 cm stem segments with terminal buds, placing on a reciprocating shaking seedbed, performing shallow layer shake culture by using a liquid adventitious bud culture medium, and performing culture by adding a growth regulator 6-BA (1.0 mg/l) and an NAA (0.05 mg/l); when the plant grows to 2.5-3.5 cm, the plant is transferred to the other half of reciprocating shaking seedbed, the shallow layer shaking culture is carried out by using a liquid rooting culture medium, and the rooting is induced by adding a growth regulator IBA (1.0 mg/l) and an IAA (1.0 mg/l).
5. The tissue culture seedling method of the Prunus cerasifera as claimed in claim 1, which is characterized in that: the reciprocating type rocking seedbed comprises a reciprocating type rocking bed and a disposable sealed clean porous seedling raising tray, the tray surface is of a plastic structure, the working mode of the reciprocating type rocking bed is horizontal reciprocating, the vibration amplitude is 20mm, and the vibration frequency is 60 rpm.
6. The tissue culture seedling method of the Prunus cerasifera as claimed in claim 1, which is characterized in that: the culture and propagation conditions are characterized in that the culture and propagation conditions are carried out by reciprocating shaking a seedbed and carrying out shallow shaking culture on a liquid culture medium at the temperature of 25 +/-3 ℃, the illumination intensity of 1000lx to 6000lx and the relative humidity of 70-80%.
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