CN101642051A - Rapid propagation technology with open tissue culture for xanthoceras sorbifolia bunge - Google Patents
Rapid propagation technology with open tissue culture for xanthoceras sorbifolia bunge Download PDFInfo
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- CN101642051A CN101642051A CN200910088122A CN200910088122A CN101642051A CN 101642051 A CN101642051 A CN 101642051A CN 200910088122 A CN200910088122 A CN 200910088122A CN 200910088122 A CN200910088122 A CN 200910088122A CN 101642051 A CN101642051 A CN 101642051A
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- 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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/22—Improving land use; Improving water use or availability; Controlling erosion
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- 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
- Y02P60/40—Afforestation or reforestation
Abstract
The invention belongs to the plant tissue culture field, and relates to a rapid propagation technology with open tissue culture for xanthoceras sorbifolia bunge. In the technology, plant tissue culture technical steps for the xanthoceras sorbifolia bunge are provided, the propagation technology with open tissue culture for the xanthoceras sorbifolia bunge is improved by an open tissue culture technology, a simple open tissue culture technology for the xanthoceras sorbifolia bunge is created, a seriously sterile operating environment, high pressure sterilization and a super clean bench are notrequired, and plant tissue culture can be carried out in a natural and open bacteria environment by adopting a plastic cup instead of a tissue culture bottle, thus fundamentally simplifying tissue culture links and reducing tissue culture cost.
Description
One, technical field
The invention belongs to the Plant Tissue Breeding field, relate to a kind of rapid propagation technology with open tissue culture of yellow horn.
Two, background technology
Yellow horn (Xanthoceras sorbifolia Bunge.) has another name called pawpaw, Wendeng City's pavilion, monk's lamp hair road, be under the jurisdiction of Sapindaceae (Sapindaceae), yellow horn belongs to (Xanthoceras), is autogenus, deciduous tree or shrub, originate in northern China, be distributed in the Qinling Mountains, to the north of the accurate river, on the south the Inner Mongol, from the east of Liaoning, to Qinghai, reach Henan and the north, Jiangsu in the south.Yellow horn comprehensive utilization value height except that saline land or ponding waterlogged lowland, can both be grown everywhere, is the pioneer tree species of planting trees on barren hills; Well developed root system, root sprout tillers are strong, are the fine tree species that environment is transformed in maintenance waterborne and sand prevention; Yellow horn seed oil content 30%, kind benevolence oil content are the distinctive woody oleiferous plants plants of a kind of China up to 60%, and the title of northern oil tea is arranged; But its seed oil is one of four kinds of woody bioenergy plants of State Administration of Forestry's recommendation through changeing esterification reaction production biodiesel; Shinyleaf yellowhorn oil is rich in unsaturated fatty acid, and nutritive value is abundant, can produce high-grade edible health care oil; Its grain of wood densification, durable is the good raw material of furniture, farm implements and artifacts; The branch leaf is used as medicine and can treats rheumatic arthritis etc., has good medical value; Designs and varieties are various and good-looking, and flowering time is longer relatively, are precious afforestation seeds and nectariferous plants; The root yellow can replace dyestuff to dye cloth, so yellow horn is with a wide range of applications and DEVELOPMENT PROSPECT.
The Biological Principles of Plant Tissue Breeding is plant cell " totipotency " and plant " palingenesis " theory.1902, the famous botanist G.Haberlanch of Germany is according to the cytology theory, and the organ and the tissue that have proposed higher plant can constantly be cut apart, up to individual cells, and plant soma has continuous division and breeding under suitable condition, develops into the viewpoint of the potentiality of whole plant.Nineteen forty-three, American White chances on and forms a bud in tobacco healing tissue cultivates, and has confirmed the argument of G.Haberlanch.Under many scientists' effort, plant tissue culture technique has obtained developing rapidly, and its theory and method are tending towards improving and are ripe, and extensive use has produced huge economic benefit and social benefit.
The numerous and diverse sport technique segment of traditional group training process is all formulated around preventing to pollute, and control has been polluted into the primary technology in the group training, can not be reduced to the acceptable scope to pollution rate, just means that also this technology can not go on.And the factor that influence is polluted is varied, as kind, concentration, disinfecting time and sterilization method, medium and the vessel of the kind of explant, the season of drawing materials, time, preprocess method, disinfectant go out the requirement of mattress, operating personnel, working environment, the work quality of superclean bench etc. all with pollute closely related, therefore, make pollution problem very complicated, also be difficult to control, become one of major reason of restriction group training industrialization.
Pollution in the group training process mainly shows as the pollution of medium, and nutritious medium provides the suitable place that grows for bacterium, fungi.Therefore, promptly guarantee the plant tissue normal growth as long as medium is transformed into, have the medium of sterilization, antibacterial functions again, in a single day mushroom loses and grows the place, just can not work the mischief to group training process.Solved pollution problem, made group training process break away from strict gnotobasis, fundamentally simplification group training link, it is fully possible carrying out open tissue culture.And the key of transforming medium is to locate a kind of broad-spectrum germicide that can add medium to.
For a long time, common antibiotics, antibiotic, preservative are done a large amount of single or combinations added test in the medium to, experimental results show that, they plant tissue culture pollute prevent and treat aspect have certain effect, but wanting to find a kind of treatment combination of broad spectrum activity at different plant varieties is the comparison difficulty, antibiotic when medium often, the growth of plant tissue will be suppressed; The plant tissue normal growth just can't obtain satisfied antibacterial effect.This also is an obstacle that domestic and international experts and scholars generally run into, that can't go beyond.Its reason one is also not have a kind of antibiotic all effective to all bacteriums, and the drug effect phase is short; The 2nd, some antibiotic, preservative are under valid density, and its sterilization, antibiotic ion pair plant tissue directly produce injury, and some then produces salt damage.At the problem in the antimicrobial agent screening, have only select with plant tissue affine, friendly, the drug effect phase long (at least one growth cycle), do not produce salt damage and have the material of sterilization, antibacterial activity, be only the only way.For this reason, we use dialectical theory of traditional Chinese medical science, follow the herbal nature compatibility monarch, minister, help, make principle, from various plants, extract and have sterilization, antibacterial substance, developed successfully that to have press down giving birth to of broad spectrum activity sterilizing ability plain, and its valid density and using method have been done a large amount of investigative tests.
The open tissue culture of plant, open, inoculate in the aseptic environments relatively, medium by additional press down to give birth to usually replace autoclaving, therefore, adding pressing down of suitable concentration in medium, to give birth to element be vital.Our evidence,, as long as transfer room is carried out disinfection comparatively completely, press down in the medium and give birth to plain concentration more than 0.06%, just can effectively prevent under the open inoculation environment, owing to be scattering into the pollution that the fungal spore on the medium causes.
Subsequently, we done press down to give birth to plain to the test of medium artificial inoculation valid density,, along with pressing down the increase of giving birth to plain concentration, the killing action of fungi is strengthened gradually, give birth to plain concentration when being 0.15% when pressing down, the fungi surviving rate is 0.But in this scope, killing effect to some bacterium is not obvious, but concentration is raised at 0.19% o'clock from 0.15%, and the survival rate of bacterium descends rapidly, illustrate that pressing down the living plain concentration of effectively killing to bacterial mass is 0.19%, determine that thus it is 0.2% that pressing down of explant inoculation medium given birth to plain concentration.
Three, summary of the invention
The present invention utilizes open tissue culture technology that the tissue culture of shinyleaf yellowhorn propagation technique is improved, created a kind of easy open yellow horn tissue culture technique, break away from strict aseptic operating environment, do not need autoclaving and superclean bench, utilize plastic cup to replace tissue culture bottle, in natural, the open tissue culture of carrying out plant in the collarium border that has, fundamentally simplification group training link has reduced group training cost.
Four, embodiment
1, initial culture (explant induction, sterile system are set up)
Get the semi-lignified stem section of yellow horn rudiment bar at fine day, cut blade, return, rinse well repeatedly with flowing water and washing powder with the plastic film bag, the stem that material is trimmed to 3--4cm is disconnected, be with 1～2 axillalry bud, put into beaker, add washing powder, after sealing with gauze, flowing water flushing 1-2h, the stem after the flushing is disconnected, be soaked in concentration and be 4% press down and give birth in the cellulose solution.Medium MS0, wherein sugared 30g/L, agar powder 4g/L, other adds the certain density living plain autoclaving that replaces that presses down.Day temperature in the culturing room (26 ± 2) ℃, nocturnal temperature (20 ± 2) ℃, one day illumination time 16h, light source adopts Incandescent fluorescent lamp.Inoculate back 7 days axillalry buds and begin swollen rising and sprout, after 25 days, it is long that axillalry bud grows to 1-2cm.
2, successive transfer culture (enrichment culture, expand numerous cultivation)
The axillalry bud of initial culture is downcut, inserts on the subculture medium, stem section originally need not, through 5-6 successive transfer culture, form the individual bud structure of growing thickly of 5-10, under 28～30 ℃, be a subculture cycle with 20 days.Subculture medium is with MS+6BA (0.8mg/L)+KT (0.8mg/L)+NAA (0.01mg/L)+sugar (30mg/L)+press down to give birth to plain (4g/L) to be advisable.In the incubation, be best with scattered light, the thick shape of sprout is crossed strong direct light seedling and is easily worn out like this, the Ruo easy excessive growth of light bud excessively, and sprout is thin and delicate, easily vitrifying.
3, culture of rootage
Aseptic bud is through continuous successive transfer culture, when reaching certain radix, can carry out culture of rootage, to obtain whole plant.Specifically more sturdy in the subculture seedling, lignification simple bud is preferably downcut, and 20 buds of a bottle graft insert on the root media, and remaining green bud grafting is gone on the subculture medium.Root media is with 1/2MS+IBA (1.5mg/L)+NAA (0.5mg/L)+sugar (20g/L)+press down to give birth to plain (4g/L).Cut when taking root bud, utilize the 1.5cm terminal bud, need not second nodal bud, such bottle seedling of taking root is neat, goes out root rate height.Going out root time and temperature has much relations, during 28 ℃ of left and right sides of temperature, and the 6th day protruding dew of root original hase, can remove to the hardening canopy and accept the natural daylight hardening this moment, about 15 days, the blade diastole, cauline leaf is slightly redly, robust plant can obviously improve a bottle seedling quality like this, bag survival rate in the raising.
4, hardening and transplanting
It is 50% natural conditions lower refining seedling 15-20 days that the test tube song that will take root moves on to degree of shading, take out tissue cultivating seedling, rinse the medium of base portion well with running water, the liquor potassic permanganate of putting into 0.1-0.5g/L soaks 1-3min, with running water flushing one time, cover wet towel and preserve moisture again in order to transplanting.To nutritious bag, the nutritious bag of matrix is housed with the disinfecting solution of potassium permanganate of 3-5g/L earlier at test-tube seedling transplanting, the test tube seedling direct transplantation after will cleaning again is in nutritious bag.The normal root water of in time drenching after the transplanting.And cover plastic film.Open plastic film after one week gradually, note the control of temperature humidity simultaneously, can reach more than 90% with this transplanting survival rate.Seedling in nutritious bag grows to 15cm and can go out land for growing field crops, garden afforestation or cultivate to adopting the fringe maternal plant.
5, the selection of culture vessel
In the traditional group training, culture vessel need select the vial of high temperature high voltage resistant and polypropylene to seal film.And in open group training, adopt additional pressing down of medium to give birth to the plain autoclaving that replaces, therefore, the range of choice of culture vessel is bigger.Among the present invention, we select for use the disposable plastic cup as culture vessel, seal with the PE preservative film.Because it is better to cultivate cup light transmission and gas permeability, plant is in cultivating cup, and robust growth is broken up and taken root and obviously is better than traditional tissue culture mode.
6, the sterilization of inoculation utensil
In the traditional group training, the scissors of inoculation usefulness, tweezers, blade etc. all adopt autoclaving or alcolhol burner calcination sterilization, and in open group training, after the inoculation utensil is cleaned with 75% alcohol, be placed on 20% and press down living cellulose solution immersion 1h, seeded process is immersed in all the time to press down and gives birth in the cellulose solution, just can effectively prevent because the pollution that the inoculation utensil causes.To autoclaving, alcolhol burner calcination sterilization with press down and give birth to the plain inoculation utensil that soaks sterilization, a large amount of tradition inoculation comparative trials have been made.The living element that presses down of high concentration has stronger bactericidal action, sterilization effect to the inoculation utensil is remarkable, the high more sterilization effect of concentration is good more, when concentration is 20%, pollution rate has dropped to 3.3%, and the living plain immersion sterilization to the inoculation utensil that presses down with 20% can effectively prevent owing to inoculate the pollution that utensil causes.
7, inoculation method
In the transfer room of relative clean, the alcohol with 75% will inoculate table top and hand is wiped clean, and opens and seal one jiao of film, with sterilized inoculation utensil, plant is inoculated into medium, and then rim of a cup obturaged get final product.
8, indoor cultivation
Place culturing rack to cultivate the vegetable material that has connected, in the incubation, regularly culturing room is sterilized, seedling is polluted in cleaning in time, keeps the culture environment of a relative clean.
9, open group training and traditional group training sport technique segment contrast and cost analysis
Autoclaving and superclean bench inoculation have been saved in open group training, have simplified group training link, and open tissue culture mode and the contrast of traditional tissue culture mode sport technique segment see that Figure of description, Figure of description are open group of training and traditional group training flow process comparison diagram.
Because pressing down, preparation gives birth to plain Chinese medicine rich material resources, production technology is simple, explant and utensil sterilized solution can reuse, take all factors into consideration open group training and traditional group training cost, comprise capital construction, testing equipment and loss, charges for water and electricity are used, the wage for workmen, environment maintenance etc., open group training is compared with the traditional group training can save cost about 60%.
Claims (9)
1, a kind of rapid propagation technology with open tissue culture of yellow horn is characterized in that may further comprise the steps:
The selection of initial culture (explant induction, sterile system set up), successive transfer culture (enrichment culture, expand numerous cultivation), culture of rootage, hardening and transplanting, culture vessel, inoculate sterilization, inoculation method, the indoor cultivation of utensil.
2, a kind of rapid propagation technology with open tissue culture of yellow horn as claimed in claim 1 when it is characterized in that initial culture, is got the semi-lignified stem section of yellow horn rudiment bar at fine day, cut blade, return, rinse well repeatedly with flowing water and washing powder with the plastic film bag, the stem that material is trimmed to 3--4cm is disconnected, be with 1～2 axillalry bud, put into beaker, add washing powder, after sealing with gauze, flowing water flushing 1-2h, the stem after the flushing is disconnected, be soaked in concentration and be 4% press down and give birth in the cellulose solution.Medium MS0, wherein sugared 30g/L, agar powder 4g/L, other adds the certain density living plain autoclaving that replaces that presses down.Day temperature in the culturing room (26 ± 2) ℃, nocturnal temperature (20 ± 2) ℃, one day illumination time 16h, light source adopts Incandescent fluorescent lamp.Inoculate back 7 days axillalry buds and begin swollen rising and sprout, after 25 days, it is long that axillalry bud grows to 1-2cm.
3, a kind of rapid propagation technology with open tissue culture of yellow horn as claimed in claim 1, when it is characterized in that successive transfer culture (enrichment culture, expand numerous cultivation), the axillalry bud of initial culture is downcut, insert on the subculture medium, stem section originally need not, through 5-6 successive transfer culture, form the individual bud structure of growing thickly of 5-10, under 28～30 ℃, be a subculture cycle with 20 days.Subculture medium is with MS+6BA (0.8mg/L)+KT (0.8mg/L)+NAA (0.01mg/L)+sugar (30mg/L)+press down to give birth to plain (4g/L) to be advisable.In the incubation, be best with scattered light, the thick shape of sprout is crossed strong direct light seedling and is easily worn out like this, the Ruo easy excessive growth of light bud excessively, and sprout is thin and delicate, easily vitrifying.
4, a kind of rapid propagation technology with open tissue culture of yellow horn as claimed in claim 1, when it is characterized in that culture of rootage, aseptic bud is through continuous successive transfer culture, when reaching certain radix, can carry out culture of rootage, to obtain whole plant.Specifically more sturdy in the subculture seedling, lignification simple bud is preferably downcut, and 20 buds of a bottle graft insert on the root media, and remaining green bud grafting is gone on the subculture medium.Root media is with 1/2MS+IBA (1.5mg/L)+NAA (0.5mg/L)+sugar (20g/L)+press down to give birth to plain (4g/L).Cut when taking root bud, utilize the 1.5cm terminal bud, need not second nodal bud, such bottle seedling of taking root is neat, goes out root rate height.Going out root time and temperature has much relations, during 28 ℃ of left and right sides of temperature, and the 6th day protruding dew of root original hase, can remove to the hardening canopy and accept the natural daylight hardening this moment, about 15 days, the blade diastole, cauline leaf is slightly redly, robust plant can obviously improve a bottle seedling quality like this, bag survival rate in the raising.
5, a kind of rapid propagation technology with open tissue culture of yellow horn as claimed in claim 1, when it is characterized in that hardening and transplanting, it is 50% natural conditions lower refining seedling 15-20 days that the test-tube plantlet that will take root moves on to degree of shading, take out tissue cultivating seedling, rinse the medium of base portion well with running water, the liquor potassic permanganate of putting into 0.1-0.5g/L soaks 1-3min, again with running water flushing one time, covers wet towel and preserves moisture in order to transplanting.To nutritious bag, the nutritious bag of matrix is housed with the disinfecting solution of potassium permanganate of 3-5g/L earlier at test-tube seedling transplanting, the test tube seedling direct transplantation after will cleaning again is in nutritious bag.The normal root water of in time drenching after the transplanting.And cover plastic film.Open plastic film after one week gradually, note the control of temperature humidity simultaneously, can reach more than 90% with this transplanting survival rate.Seedling in nutritious bag grows to 15cm and can go out land for growing field crops, garden afforestation or cultivate to adopting the fringe maternal plant.
6, a kind of rapid propagation technology with open tissue culture of yellow horn as claimed in claim 1 is characterized in that the selection of culture vessel, and in the traditional group training, culture vessel need select the vial of high temperature high voltage resistant and polypropylene to seal film.And in open group training, adopt additional pressing down of medium to give birth to the plain autoclaving that replaces, therefore, the range of choice of culture vessel is bigger.Among the present invention, we select for use the disposable plastic cup as culture vessel, seal with the PE preservative film.Because it is better to cultivate cup light transmission and gas permeability, plant is in cultivating cup, and robust growth is broken up and taken root and obviously is better than traditional tissue culture mode.
7, a kind of rapid propagation technology with open tissue culture of yellow horn as claimed in claim 1, it is characterized in that inoculating the sterilization of utensil, in the traditional group training, the scissors of inoculation usefulness, tweezers, blade etc. all adopt autoclaving or alcolhol burner calcination sterilization, and in open group training, after the inoculation utensil was cleaned with 75% alcohol, being placed on 20% pressed down and gives birth to cellulose solution immersion 1h, seeded process is immersed in all the time to press down and gives birth in the cellulose solution, just can effectively prevent because the pollution that the inoculation utensil causes.To autoclaving, alcolhol burner calcination sterilization with press down and give birth to the plain inoculation utensil that soaks sterilization, a large amount of tradition inoculation comparative trials have been made.The living element that presses down of high concentration has stronger bactericidal action, sterilization effect to the inoculation utensil is remarkable, the high more sterilization effect of concentration is good more, when concentration is 20%, pollution rate has dropped to 3.3%, and the living plain immersion sterilization to the inoculation utensil that presses down with 20% can effectively prevent owing to inoculate the pollution that utensil causes.
8, a kind of rapid propagation technology with open tissue culture of yellow horn as claimed in claim 1, it is characterized in that inoculation method, in the transfer room of relative clean, alcohol with 75% will inoculate table top and hand is wiped clean, open and seal one jiao of film, with sterilized inoculation utensil, plant is inoculated into medium, and then rim of a cup obturaged gets final product.
9, a kind of rapid propagation technology with open tissue culture of yellow horn as claimed in claim 1, it is characterized in that the indoor cultivation method, place culturing rack to cultivate the vegetable material that has connected, in the incubation, regularly culturing room is sterilized, seedling is polluted in cleaning in time, keeps the culture environment of a relative clean.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102217543A (en) * | 2011-05-24 | 2011-10-19 | 崔刚 | Open plant tissue culture and factory rapid propagation method |
| CN102217547A (en) * | 2011-06-01 | 2011-10-19 | 甘肃农业大学 | Opening sealing method of glass culture vessel for callus culture |
| CN104285816A (en) * | 2014-11-03 | 2015-01-21 | 杨业容 | Rapid propagation method for xanthoceras sorbifolia bunge tissue during culturing |
| CN104322376A (en) * | 2014-11-25 | 2015-02-04 | 张勇 | Cultivation method of stem tissues of shiny-leaved yellowhorn |
| CN104604680A (en) * | 2015-01-19 | 2015-05-13 | 杭州雷威农业开发有限公司 | Growth medium capable of promoting seed germination and seedling growth of bletilla striata, formula of growth medium and preparation method |
| CN105580732A (en) * | 2015-12-11 | 2016-05-18 | 大连民族大学 | Shiny-leaved yellowhorn tissue culture rapid propagation method with root as explant and shiny-leaved yellowhorn adventitious bud cultured through method |
| CN106993532A (en) * | 2017-03-31 | 2017-08-01 | 中国林业科学研究院热带林业研究所 | A kind of open tissue culture method of yearning between lovers |
| CN113545292A (en) * | 2021-08-18 | 2021-10-26 | 山东丰沃农业有限公司 | Culture medium combination and culture method of xanthoceras sorbifolia seedling regeneration system |
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2009
- 2009-07-08 CN CN200910088122A patent/CN101642051A/en active Pending
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102217543A (en) * | 2011-05-24 | 2011-10-19 | 崔刚 | Open plant tissue culture and factory rapid propagation method |
| CN102217547A (en) * | 2011-06-01 | 2011-10-19 | 甘肃农业大学 | Opening sealing method of glass culture vessel for callus culture |
| CN104285816A (en) * | 2014-11-03 | 2015-01-21 | 杨业容 | Rapid propagation method for xanthoceras sorbifolia bunge tissue during culturing |
| CN104322376A (en) * | 2014-11-25 | 2015-02-04 | 张勇 | Cultivation method of stem tissues of shiny-leaved yellowhorn |
| CN104604680A (en) * | 2015-01-19 | 2015-05-13 | 杭州雷威农业开发有限公司 | Growth medium capable of promoting seed germination and seedling growth of bletilla striata, formula of growth medium and preparation method |
| CN105580732A (en) * | 2015-12-11 | 2016-05-18 | 大连民族大学 | Shiny-leaved yellowhorn tissue culture rapid propagation method with root as explant and shiny-leaved yellowhorn adventitious bud cultured through method |
| CN105580732B (en) * | 2015-12-11 | 2017-10-03 | 大连民族大学 | A kind of shiny-leaved yellowhorn tissue cultivation rapid breeding method and its shiny-leaved yellowhorn adventitious bud of culture for explant |
| CN106993532A (en) * | 2017-03-31 | 2017-08-01 | 中国林业科学研究院热带林业研究所 | A kind of open tissue culture method of yearning between lovers |
| CN106993532B (en) * | 2017-03-31 | 2019-10-22 | 中国林业科学研究院热带林业研究所 | A kind of open tissue culture method of yearning between lovers |
| CN113545292A (en) * | 2021-08-18 | 2021-10-26 | 山东丰沃农业有限公司 | Culture medium combination and culture method of xanthoceras sorbifolia seedling regeneration system |
| CN113545292B (en) * | 2021-08-18 | 2022-04-29 | 山东丰沃农业有限公司 | Culture medium combination and culture method of xanthoceras sorbifolia seedling regeneration system |
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