NL2034490A - Shoot Tip Culture of Strawberry Without Hormone - Google Patents

Shoot Tip Culture of Strawberry Without Hormone Download PDF

Info

Publication number
NL2034490A
NL2034490A NL2034490A NL2034490A NL2034490A NL 2034490 A NL2034490 A NL 2034490A NL 2034490 A NL2034490 A NL 2034490A NL 2034490 A NL2034490 A NL 2034490A NL 2034490 A NL2034490 A NL 2034490A
Authority
NL
Netherlands
Prior art keywords
strawberry
hormone
culture
hours
explants
Prior art date
Application number
NL2034490A
Other languages
Dutch (nl)
Inventor
Zhang Yujuan
He Jinyu
Du Xiaoqiu
Liu Lisha
Wang Mei
Li Defu
Original Assignee
Nanchong Acad Of Agricultural Sciences
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Nanchong Acad Of Agricultural Sciences filed Critical Nanchong Acad Of Agricultural Sciences
Publication of NL2034490A publication Critical patent/NL2034490A/en

Links

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
    • 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
    • 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
    • A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
    • A01H6/74—Rosaceae, e.g. strawberry, apple, almonds, pear, rose, blackberries or raspberries
    • A01H6/7409—Fragaria, i.e. strawberries
    • 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)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Botany (AREA)
  • Environmental Sciences (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

A shoot tip culture of strawberry without hormone is carried out in the following steps: (1) Preparation of MT liquid medium solution with a mass concentration of sucrose of 7-8%; (2) Adding liquid medium solution into the petridish to make liquid medium; (3) Strawberry stolon terminal buds of 4-6cm were clipped, the outer large leaves were peeled off, and explants were obtained by washing. (4) The explants were first disinfected with alcohol, then disinfected with sodium hypochlorite solution, and then washed with sterile water; (5) The leaves of the sterile explants were stripped and the shoot tips with a length of 0.3-0.5mm were cut; (6) lnoculated into liquid medium, and then placed in incubator for primary culture; (7) The shoot tips were transferred to MS solid medium and cultured until clumping buds and roots were differentiated. The method of the invention does not need to be limited by hormone concentration; Under high sugar concentration, the shoot tips were not easy to brown and the growth rate was moderate, which ensured the genetic stability of the varieties.

Description

Nanchong Academy of Agricultural Sciences 23/030 NL
Shoot Tip Culture of Strawberry Without Hormone
Technical Field
The invention belongs to the technical field of plant culture, in particular to a hormone- free culture method for strawberry shoot tips.
Background Technology
Shoot tip culture is widely used in rapid propagation of seedlings, virus-free seedling production and variety improvement. The plant is genetically stable after shoot tip tissue culture, which is especially suitable for species with difficulty in conventional breeding, trees with long childhood or species requiring detoxification. In 1960, Morel was the first to breed orchids from the shoot tips. By tissue culture of the shoot tips, he obtained virus-free plants, thus overcoming the problems of low seed propagation efficiency and variety degradation of orchids and promoting the prosperity of the orchid industry.
At present, MS medium or slightly modified medium is commonly used in shoot tip culture, and other substances such as nutrients and growth regulating substances are added to improve the seedling formation rate of shoot tip tissue culture. The carbon source of MS medium is usually 2-4% glucose or sucrose. The types and concentrations of plant hormones vary with different plant varieties. NAA is the most commonly used auxin, followed by IAA. Gibberellin is beneficial to shoot tip elongation and survival, requiring a low concentration, and too high concentration will have adverse effects.
The survival rate of shoot tip culture was positively correlated with shoot tip size. Shoot tips with the size of a few millimeters are preferred for rapid propagation, while shoot tip growth points smaller than 0.5mm are required for virus-free seedling culture. Shoot tip growth points refer to growth cones with 1-2 leaf primordiums. Diseases caused by viruses are different from fungal and bacterial diseases, which cannot be controlled by fungicides and antibiotics. The virus is not evenly distributed in the plant body. Because there is no vascular bundle, the virus can only be transmitted through the plasmata, which can not catch up with the speed of cell division and growth, so the growth point of the shoot tip almost contains no virus. shoot tip culture has good virus-free effect and stable offspring, which is widely used now. shoot tip culture of strawberry, which started in 1960s, can not only accelerate the propagation of excellent strawberry varieties for commercial production in a short time, but also save the excellent strawberry varieties seriously infected with virus.
Strawberry shoot tip culture usually uses MS medium combined with different concentrations of hormones. However, because different varieties of shoot tips have different demands on hormone types and concentrations, operators have to constantly explore and adjust hormone types and concentrations for shoot tip culture, which is time-consuming and laborious, which is not good for large-scale popularization.
Summary of the Invention
The purpose of the invention is to provide a hormone-free culture method for strawberry shoot tips. The high sugar liquid medium without any hormone is used to culture strawberry shoot tips, and strawberry seedlings can be directly obtained by cultivation of strawberry shoot tips.
The method of the invention is carried out according to the following steps: (1) MT (Murashige and Tucker) liquid culture-medium solution was prepared with sucrose concentration of 7-8%. (2) Add liquid medium solution to the petri dish, make liquid medium, seal with sealing film for use; (3) Strawberry stolon terminal buds of 4-6cm were clipped, the outer large leaves were peeled off, and explants were obtained by washing. (4) The explants were disinfected with alcohol for 20-30 seconds, then disinfected with sodium hypochlorite solution for 15-25 minutes, and then washed with sterile water to obtain sterile explants; (5) The stereopter was placed in an ultra-clean table, and the leaves of the sterile explants were peeled off under the stereopter, and the shoot tips with a length of 0.3-0.5mm were cut with a scalpel; (6) The shoot tips were inoculated into liquid medium and placed in an incubator for primary culture for 2-3 weeks; (7) After completion of primary culture, the obtained cultured shoot tips were transferred to a triangular flask containing MS solid medium, and then cultured in an incubator until clumping buds and roots were differentiated.
In Step (1) above, after sterilization, it is divided into 100ml/bottle.
In Step (2) above, the petri dish was a 60mm diameter glass petri dish after sterilization.
In Step (2) above, adding liquid medium solution to Petri dishes is adding liquid medium solution to several petri dishes on a super clean workbench.
In Step (2) above, 3ml was added to each petri dish.
In Step (3) above, washing means washing with running water.
In Step (4) above, the mass concentration of alcohol ranges from 70-75%.
In Step (4) above, the mass concentration of sodium hypochlorite solution is 1-1.5%.
In Step (4) above, sterile water was cleaned 4-5 times.
In Step (4) above, sterile explants were placed in petri dishes lined with filter paper before stripping the growth points of shoot tips.
In Step (6) above, the petri dish was placed horizontally on the separator of the incubator. The culture conditions of the incubator were as follows: The light intensity is 1600- 2200Lux, there are 16 hours of 40+0.5°C light and 8 hours of 30+0.5°C darkness within 24 hours; The 40+0.5°C treatment was used to passivate the virus
In Step (7) above, 30mLMS solid medium was added into each 100mL triangular bottle, and MS solid medium was inserted into the base of shoot tips after culture, so that the shoot tips after culture were continued to be cultured in a sterile environment. The culture conditions of the incubator were as follows: The culture condition of the incubator is: The light intensity is 1600-2200Lux, there are 16 hours of 25+0.5°C light and 8 hours of 25+0.5°C darkness within 24 hours.
The method of the invention does not need to be limited by hormone concentration;
The invention uses high sugar MT medium. Under high sugar concentration, the shoot tips are not easy to brown, the growth rate is moderate, and the callus will not be generated. The passivation of the virus at 40+0.5°C does not affect the growth of the shoot tips, ensuring the genetic stability of the varieties.
The invention can obtain regenerated seedlings of the three strawberry varieties of
Red Face Strawberry, Sweet Charlie Strawberry and Cream Strawberry, and significantly improve the efficiency of plant shoot tip culture, indicating that the medium is suitable for multiple strawberry varieties and can be widely used in different strawberry varieties in the future.
The main advantages of the invention are: (I) Do not need to be limited by hormone concentration:
Strawberry shoot tip culture usually uses MS medium combined with different concentrations of hormones. However, because different varieties of shoot tips have different demands on hormone types and concentrations, operators have to constantly explore and adjust hormone types and concentrations for shoot tip culture, which is time- consuming and laborious, which is not conducive to large-scale promotion. The invention uses high sugar MT medium, under high sugar concentration, the shoot tip is not easy to brown, the growth rate is moderate, and the callus will not be generated, thus ensuring the genetic stability of the variety. (Il) Virus can be passivated at the same time during shoot tip culture, which can significantly save production cost:
In previous studies, virus-free strawberry was treated by selecting healthy plants with no obvious disease and pure varieties. Plants were planted in a pot and treated at 40°C for 16 h, 35°C for 8 h and variable temperature for 4 weeks in an artificial climate chamber.
Then, shoot tip stripping and shoot tip culture were carried out on the new shoots. Heat treatment in the artificial climate chamber required a large amount of energy and high cost due to the large number of plants treated. Strawberry shoot tips below 0.5mm were cut and placed in 60mm petri dish with only 3ml liquid medium. Heat treatment was carried out in ordinary incubators. A 200L capacity incubator could process hundreds of different samples at the same time. A large amount of saving hardware facilities and operation costs, more in line with the current environmental requirements of energy conservation and emission reduction; (lll) Application Prospect:
Currently, MS medium + plant hormone is commonly used for strawberry shoot tip culture. The hormone types and concentrations of different varieties are not the same, which requires the operator to spend a lot of energy to explore the hormone formula for different varieties. The invention adopts high sugar MT medium for shoot tip culture of three strawberry varieties, namely, Red Face Strawberry, Sweet Charlie Strawberry and Cream
Strawberry, without using hormones, and can effectively obtain regenerated seedlings, and significantly improve the shoot tip culture efficiency of strawberry. The passivation virus was treated with day-night alternating high temperature during shoot tip culture, which saved the production unit of strawberry tissue culture seedlings from buying artificial climate chamber and saving a lot of power consumption. This method is beneficial to the large-scale production of strawberry tissue culture.
Brief Description of the Drawings
Figure 1 is a photograph of the shoot tip in Implementation method 1 of the invention and the appearance of the cultured shoot tip after 15 days of primary culture; In the figure, the left is the growth point of the shoot tip, and the right is the shoot tip after culture after 15 days of primary culture.
Figure 2 is a photograph of the appearance of a regenerated strawberry seedling from culture to differentiation into clumped buds and roots in Implementation method 1 of the invention.
Specific Implementation Methods
The varieties of strawberries used in the Implementation methods of the invention are
Red Face Strawberry, Sweet Charlie Strawberry and Cream Strawberry.
In the implementation methods of the invention, the survival rate of culture is more than 95%.
Implementation method 1
MT liquid medium solution was prepared with sucrose concentration of 7%. After sterilization, it was divided into 100ml/ bottle.
The liquid medium solution was added into several petri dishes on the ultra-clean 5 workbench, and the amount of each dish was 3ml. The liquid medium was prepared and sealed with a sealing film for later use. The petri dish was a 60mm diameter glass petri dish after sterilization.
Strawberry stolon top buds of 5cm were clipped, the outer large leaves were peeled off, and the explants were obtained by washing with running water.
The explants were disinfected with alcohol for 25s, then disinfected with sodium hypochlorite solution for 20min, and then washed with sterile water to obtain sterile explants.
The mass concentration of alcohol is 73%; The mass concentration of sodium hypochlorite solution was 1.3%. Sterile water cleaning times of 5 times; Place in a petri dish lined with filter paper and set aside.
The stereopter was placed in an ultra-clean table. After the sterile explants were stripped off under the stereopter, the shoot tip with a length of 0.5mm was cut with a scalpel under the stereopter.
The shoot tips were inoculated into liquid medium and placed in an incubator for primary culture for 15 days. The light intensity is 1600-2200Lux, there are 16 hours of 40+0.5°C light and 8 hours of 30+0.5°C darkness within 24 hours; The 40+0.5°C treatment was used to passivate the virus.
The appearance photos of shoot tips and cultured shoot tips were shown in Figure 1.
The cultured shoot tips were transferred to triangular bottles containing MS solid medium and cultured until clumping buds and roots were differentiated. Culture conditions:
The light intensity is 1600-2200Lux, there are 16 hours of 25+0.5°C light and 8 hours of 2510.5°C darkness within 24 hours; The appearance photos of regenerated strawberry seedlings obtained are shown in Figure 2.
Implementation method 2
The method is the same as that of Implementation method 1, with differences in: (1) Sucrose concentration in MT liquid medium solution was 8%; (2) Strawberry stolon terminal buds of 4cm were clipped; (3) The explants were disinfected with alcohol for 20s, then disinfected with sodium hypochlorite solution for 15min, and then washed with sterile water to obtain sterile explants;
The mass concentration of alcohol is 75%; The mass concentration of sodium hypochlorite solution was 1.5%. Sterile water cleaning times of 4 times; (4) Cut the shoot tip with a length of 0.4mm;
(5) Put them in the incubator for primary culture for 2 weeks; The light intensity of the incubator is 1600Lux; (6) The light intensity of the incubator was 1600Lux whn the tufted buds and roots were differentiated.
Implementation method 3
The method is the same as that of Implementation method 1, with differences in: (1) The sucrose concentration in MT liquid medium solution was 7.5%; (2) Strawberry stolon terminal buds of 6cm were clipped; (3) The explants were disinfected with alcohol for 30s, then disinfected with sodium hypochlorite solution for 25min, and then washed with sterile water to obtain sterile explants;
The mass concentration of alcohol is 70%; The mass concentration of sodium hypochlorite solution was 1%. Sterile water cleaning times of 4 times; (4) Cut the shoot tip with a length of 0.3mm; (5) Put them in the incubator for primary culture for 3 weeks; The light intensity of the incubator is 2200Lux; (6) The light intensity of the incubator was 2200Lux when the tufted buds and roots were differentiated.

Claims (7)

ConclusiesConclusions 1. Werkwijze voor het hormoonvrij kweken van stengeltoppen van aarbeien, met het kenmerk, dat deze de volgende stappen omvat: (1) het bereiden van een oplossing van MT kweekmedium met een sucrose- concentratie van 7-8%, die wordt gesteriliseerd en opnieuw wordt samengevoegd; (2) het toevoegen van de oplossing van het kweekmedium aan een petrischaal, waardoor een vloeibaar kweekmedium wordt bereid, dat wordt afgedicht met afdichtfolie voor later gebruik; (3) het knippen van aardbeistengeltoppen met een lengte van 4-6 cm, het verwijderen van de grote buitenste bladeren en het wassen van de stekken om explantaten te verkrijgen; (4) het desinfecteren van de explantaten met alcohol gedurende 20-30 seconden, vervolgens het desinfecteren met een oplossing van natriumhypochloriet gedurende 15-25 minuten en vervolgens het wassen met steriel water om steriele explantaten te verkrijgen; (5) het plaatsen van een dissectiemcroscoop (“stereopter’) in een laminaire flowkast (“ultra-clean table”) en het onder de dissectiemicroscoop verwijderen van de buitenste bladeren van het steriele explantaat, en het snijden van de stengeltoppen met een lengte van 0,3-0,5 mm met een dissectiemes; (6) het enten van de stengeltoppen in vloeibaar medium en het plaatsen in een incubator voor primaire kweek gedurende 2-3 weken; (7) het overbrengen van de verkregen gekweekte stengeltoppen na voltooiing van de primaire kweek naar een driehoekige fles met vast MS kweekmedium en vervolgens het kweken daarvan in een incubator om tot kiemknoppen en wortels te differentiëren.A method for cultivating strawberry stem tops without hormones, characterized in that it comprises the following steps: (1) preparing a solution of MT culture medium having a sucrose concentration of 7-8%, which is sterilized and reconstituted is merged; (2) adding the culture medium solution to a petri dish, thereby preparing a liquid culture medium, which is sealed with sealing film for later use; (3) clipping strawberry stem tips with a length of 4-6 cm, removing the large outer leaves and washing the cuttings to obtain explants; (4) disinfecting the explants with alcohol for 20-30 seconds, then disinfecting them with a solution of sodium hypochlorite for 15-25 minutes, and then washing them with sterile water to obtain sterile explants; (5) placing a dissecting microscope (“stereopter”) in a laminar flow cabinet (“ultra-clean table”) and removing the outer leaves of the sterile explant under the dissecting microscope, and cutting the stem tips to a length of 0.3-0.5 mm with a dissecting blade; (6) inoculating the stem tips in liquid medium and placing in a primary culture incubator for 2-3 weeks; (7) transferring the obtained grown stem tips after completion of the primary culture to a triangular flask of solid MS culture medium and then culturing it in an incubator to differentiate into buds and roots. 2. Werkwijze voor het hormoonvrij kweken van stengeltoppen van aarbeien volgens conclusie 1, met het kenmerk, dat in stap (2) de toegevoegde hoeveelheid aan iedere petrischaal 3 ml bedraagt.A method for the hormone-free cultivation of strawberry stem tops according to claim 1, characterized in that in step (2) the amount added to each petri dish is 3 ml. 3. Werkwijze voor het hormoonvrij kweken van stengeltoppen van aarbeien volgens conclusie 1, met het kenmerk, dat in stap (4) de massaconcentratie van de alcohol 70-75% bedraagt.A method for the hormone-free cultivation of strawberry stem tops according to claim 1, characterized in that in step (4) the mass concentration of the alcohol is 70-75%. 4. Werkwijze voor het hormoonvrij kweken van stengeltoppen van aarbeien volgens conclusie 1, met het kenmerk, dat in stap (4) de massaconcentratie van de natrium- hypochlorietoplossing 1-1.5% bedraagt.A method for the hormone-free cultivation of strawberry stem tops according to claim 1, characterized in that in step (4) the mass concentration of the sodium hypochlorite solution is 1-1.5%. 5. Werkwijze voor het harmoonvrij kweken van stengeltoppen van aarbeien volgens conclusie 1, met het kenmerk, dat in stap (4) de frequentie van het spoelen met steriel water 4-5 keer bedraagt.A method for the harmon-free cultivation of strawberry stem tops according to claim 1, characterized in that in step (4) the frequency of rinsing with sterile water is 4-5 times. 6. Werkwijze voor het hormoonvrij kweken van stengeltoppen van aarbeien volgens conclusie 1, met het kenmerk, dat in stap (6) de kweekomstandigheden van de incubator zijn: een lichtintensiteit van 1600-2200 Lux, 16 uur licht bij 40 + 0.5°C en 8 uur duisternis bij 30 £ 0.5°C binnen 24 uur; waarbij de behandeling bij 40 + 0.5°C wordt toegepast om virussen te deactiveren.A method for the hormone-free cultivation of strawberry stem tops according to claim 1, characterized in that in step (6) the cultivation conditions of the incubator are: a light intensity of 1600-2200 Lux, 16 hours of light at 40 + 0.5°C and 8 hours of darkness at 30 £ 0.5°C within 24 hours; wherein the treatment at 40 + 0.5°C is used to inactivate viruses. 7. Werkwijze voor het hormoonvrij kweken van stengeltoppen van aarbeien volgens conclusie 1, met het kenmerk, dat in stap (7) de kweekomstandigheden van de incubator zijn: een lichtintensiteit van 1600-2200 Lux, 16 uur licht bij 25 + 0.5°C en 8 uur duisternis bij 25 + 0.5°C binnen 24 uur.A method for the hormone-free cultivation of strawberry stem tops according to claim 1, characterized in that in step (7) the cultivation conditions of the incubator are: a light intensity of 1600-2200 Lux, 16 hours of light at 25 + 0.5°C and 8 hours of darkness at 25 + 0.5°C within 24 hours.
NL2034490A 2022-04-07 2023-04-03 Shoot Tip Culture of Strawberry Without Hormone NL2034490A (en)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202210359341.2A CN114788495B (en) 2022-04-07 2022-04-07 Hormone-free strawberry stem tip culture method

Publications (1)

Publication Number Publication Date
NL2034490A true NL2034490A (en) 2023-04-26

Family

ID=82462580

Family Applications (1)

Application Number Title Priority Date Filing Date
NL2034490A NL2034490A (en) 2022-04-07 2023-04-03 Shoot Tip Culture of Strawberry Without Hormone

Country Status (2)

Country Link
CN (1) CN114788495B (en)
NL (1) NL2034490A (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116686707A (en) * 2023-06-12 2023-09-05 江苏省农业科学院 Pretreatment method for plant stem tip tissue culture

Family Cites Families (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2996995B2 (en) * 1988-06-01 2000-01-11 ザ テキサス エイ アンド エム ユニヴァーシティ システム Transformation method of plant by shoot tip
JPH0690623A (en) * 1991-08-29 1994-04-05 Nippon Chemitec Kk Shoot tip grafting culture of oriental orchid
JPH05260869A (en) * 1992-02-10 1993-10-12 Iseki & Co Ltd Adventitious root of sweet potato, its inducing method and its culture method
CN100420369C (en) * 2006-04-30 2008-09-24 江汉大学 Method for obtaining hormone-free tissue culture detoxified seedling of chrysanthemum for tea use
CN102613086B (en) * 2012-03-31 2013-12-11 南京农业大学 Hormone-free tissue culture method for dendrobium candidum
CN103283595B (en) * 2013-05-17 2015-04-22 高红胜 Primary culture method of stem tip tissue of strawberry
CN103314863B (en) * 2013-07-16 2016-01-20 方倩 A kind of method of efficient acquisition tulip callus
CN103535277A (en) * 2013-09-18 2014-01-29 安徽省农业科学院园艺研究所 Method for efficiently producing clocasia escalenta Schott microsphere stems
CN103975858A (en) * 2014-05-29 2014-08-13 天津滨城天龙农业科技有限公司 Hormone-free culture medium for culturing high-quality butterfly orchid seedlings and application of hormone-free culture medium
CN113584072B (en) * 2021-07-29 2024-02-06 上海市农业科学院 Construction method of strawberry genetic transformation system

Also Published As

Publication number Publication date
CN114788495A (en) 2022-07-26
CN114788495B (en) 2023-04-14

Similar Documents

Publication Publication Date Title
CN102150624B (en) Tissue culture and rapid propagation method for pinellia tuber plant
CN111616052A (en) Rapid propagation and sugar-free rooting culture method and application of apple rootstock catalpa bungei
CN105052528A (en) Industrialized ramie seedling culturing method
CN110352853A (en) A method of improving Atractylis lancea test tube seedling quality and transplanting survival rate
CN102388800B (en) Light source control method for tissue culture of brassica napus
CN107155898A (en) A kind of dendrobium candidum carries out expanding numerous method using stem section thin slice
CN109220810A (en) A kind of method of tree peony embryo high-effective root-growing under aseptic condition
CN105265316B (en) A kind of allium plateau rapid propagation method
NL2034490A (en) Shoot Tip Culture of Strawberry Without Hormone
CN101695280B (en) Tissue culture and rapid propagation method of raspberries
CN101828527B (en) Method for controlling quick propagation light source for upland cotton
CN110476818A (en) A kind of auxiliary method for promoting tung oil tree stem with bud regeneration plant of physics
CN105475129A (en) Tissue-culture rapid propagation method for arundina graminifolia
US20230397559A1 (en) Method of hormone-free shoot tip culture for citrus
CN109042330A (en) A kind of method for tissue culture of spindle tree
CN104719168A (en) Method for culturing bletilla striata seedling by virtue of intermittent immersion type bioreactor
CN110476807B (en) Method for establishing sterile culture system of mature seed embryo of peony 'paeonia ostii' for oil
CN114557279B (en) Paphiopedilum purpureum non-symbiotic germination culture medium and culture method
CN100403882C (en) Cultivation of peony germinal dislocation
CN107683771A (en) A kind of method of the efficient sterile culture of tree peony embryo
CN106305419B (en) A kind of method of open culture sugar-cane tissue culture seedlings
CN104115748A (en) Culture method for rapid propagation of cotinus coggygria 'royal purple'
CN114467753B (en) Tissue culture method of silvery deer maple
CN111642396B (en) Culture medium and culture method suitable for linum sarmentosum anther and ovary
CN111972074B (en) Seed treatment method for early-maturing sweet cherry variety and method for sowing seedlings in current year