CN110432148A - Extend the method for carrot callus' storage life - Google Patents
Extend the method for carrot callus' storage life Download PDFInfo
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- CN110432148A CN110432148A CN201910827865.8A CN201910827865A CN110432148A CN 110432148 A CN110432148 A CN 110432148A CN 201910827865 A CN201910827865 A CN 201910827865A CN 110432148 A CN110432148 A CN 110432148A
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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
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- Developmental Biology & Embryology (AREA)
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Abstract
The invention discloses a kind of methods for extending carrot callus' storage life, comprising the following steps: step 1: choosing after current year raw carrot root tuber pre-processes, stripping and slicing is inoculated in induced medium, in dark condition culture to growing callus；Step 2: the callus grown being inoculated in and is saved in subculture medium, is placed under low temperature or condition of ultralow temperature and saves.A kind of method that carrot callus' recovery regrowth is also disclosed, the carrot callus that will be saved under low temperature or condition of ultralow temperature are inoculated on different recovery subculture mediums respectively, are cultivated under the conditions of 25-27 DEG C.It recovers again after callus low temperature of the invention or cryopreservation, storage life can be extended, reduce the number of squamous subculture, moreover it is possible to the relative growth rate after improving recovery.
Description
Technical field
The present invention relates to Plant Tissue Breeding and germplasm resource Techniques of preserving field, in particular to a kind of extension carrot is cured
The method of injured tissue storage life.
Background technique
Carrot (scientific name: Daucus carota L.var.sativa Hoffm) belongs to Umbelliferae 1 year or biennial herb
Plant, root thickness are strong, conico-acuminate, in orange red.It is one of global ten big vegetable crops, adaptable, is easily cultivated, plantation
It is very universal, it is dispersed throughout Asia, the multiple areas in Europe and America.Carrotene in carrot is the main source of vitamin A,
It can promote to grow, prevent bacterium infection, and there is protection epidermal tissue, on protection respiratory tract, alimentary canal, urinary system are planned as a whole
The function and effect of skin cell tissue；Carrot also contains Quercetin, and coronary blood flow can be increased by often eating, and promotes adrenal gland
Element synthesis, has the effect of decompression, anti-inflammatory, in view of the important nutritive value of carrot, people also have become its growing interest
One of the object of scientific research personnel's research, in recent years, scientific research personnel obtains carrot culture by way of Plant Tissue Breeding
Seedling or breeding material etc. are no longer rare.
The tissue cultures broad sense of plant is called in vitro culture, refer to isolated from plant the tissue to suit the requirements, organ or
Cell, protoplast etc. are cultivated under manual control condition by sterile working to obtain regenerated intact plant or life
Produce the technology with other products of economic value.Narrow sense refers to plant parts tissue, such as forming layer, parenchymal tissue, leaf
Meat tissue, root tuber etc. are cultivated, to obtain the technology of other products of regenerated intact plant or production with economic value.
But the basis for successfully carrying out Plant Tissue Breeding is and the existing callus there are high-quality and abundance callus
Source is to carry out saving by continuous squamous subculture getting mostly, and existing callus is all regular subculture when saving,
Heavy workload, and subculture number excessively also results in the varied in ploidy of material, is unfavorable for providing good callus at any time
To produce a large amount of benign systems, monoploid, polyploid or as scientific research material etc., and the preservation of existing carrot callus
It is no exception, perplexed by same technical problem.Therefore, how to extend the storage life of callus, reduce periodically after generation
Several and callus varied in ploidy probability, to solving, existing issue is very necessary.
Summary of the invention
It is an object of the invention to solve at least the above problems, and provide the advantages of at least will be described later.
It is a still further object of the present invention to provide a kind of methods for extending carrot callus' storage life, by low temperature or
Cryopreservation can extend the time of squamous subculture, reduce the number of regular subculture.
It is a still further object of the present invention to provide a kind of method of carrot callus' recovery regrowth, for low temperature or
The callus of cryopreservation, can on recovery subculture medium normal restoration ecosystem.
In order to realize object of the present invention and further advantage, a kind of extension carrot callus storage life is provided
Method, comprising the following steps:
Step 1: after choosing current year raw carrot root tuber pretreatment, stripping and slicing, then the carrot explant of stripping and slicing is inoculated in
Induced medium, in dark condition culture to growing callus；The induced medium be containing 0.1-0.2mg/L KT,
The MS culture medium of 0.2mg/L 2,4-D, 25-30g/L sucrose and 6.5-7.5g/L agar；
Step 2: the callus grown being inoculated in and is saved in subculture medium, low temperature or condition of ultralow temperature are placed in
Lower preservation, the preservation subculture medium be containing 0.1-0.2mg/L KT, 2 0.1-0.2mg/L, 4-D, 10-20g/L sucrose,
The MS culture medium of 25-30g/L mannitol and 50 μm of ol/L ABA, wherein when cryo-conservation, in the preservation subculture medium
On the basis of by 6.5-7.5g/L add agar.
Preferably, carrot selected in step 1 is the fresh Hu without wound, healthy growth uniformly and within harvesting 2 weeks
Radish, preservation under the conditions of 4 DEG C after harvesting；
Carrot preprocess method are as follows: will choose carrot clean after, with mass fraction be 75% alcoholic solution impregnate 30-
40s, then with mass fraction be 0.1%-0.2% mercuric chloride solution impregnate 10-15min, later use aseptic water washing 4-6 times, be placed in
It absorbs water on aseptic paper.
Preferably, cultivation temperature is 25-27 DEG C in step 1.
Preferably, low temperature is 4 DEG C；Ultralow temperature is -20 DEG C or -80 DEG C.
The present invention also provides a kind of method of carrot callus' recovery regrowth, the carrot callus are to adopt
The carrot callus saved with the method, the method for carrot callus' recovery regrowth specifically: will
The carrot callus saved under low temperature or condition of ultralow temperature are inoculated on different recovery subculture mediums, 25-27 respectively
It is cultivated under the conditions of DEG C.
Preferably, the recovery subculture medium of the carrot callus of cryo-conservation be containing 0.2mg/L KT,
The MS culture medium of 0.2-0.5mg/L 2,4-D, sucrose 20-30g/L and agar 6.5-7.5g/L；
The carrot callus of cryopreservation are first transferred to liquid rejuvenation culture medium according to method is melted fastly after defrosting
On, then be transferred in solid rejuvenation culture medium and cultivate, restoration ecosystem；The liquid rejuvenation culture medium be by fluid nutrient medium with it is sweet
0.4-1:1 is mixed oil by volume, the fluid nutrient medium be containing 0.2-0.5mg/L KT, 1.0mg/L NAA and
The MS culture medium of sucrose 20-30g/L；The solid rejuvenation culture medium is to contain 0.2-0.5mg/L KT, 1.0mg/L NAA, sugarcane
The MS culture medium of sugared 20-30g/L and agar 6.5-7.5g/L.
It is preferably, described to melt method fastly, specifically: the callus being inoculated into the liquid rejuvenation culture medium, first
It is put into 4 DEG C of refrigerators 6 hours, places into -20 DEG C of refrigerator overnights, be put into -80 DEG C of refrigerator long-term preservations later, taking-up when needing recovery
Afterwards, it is sequentially placed in 30 DEG C and 35 DEG C of water-bath each half an hour, completes to thaw.
The present invention is include at least the following beneficial effects:
For the present invention using the root tuber of carrot as source, Fiber differentiation grows up to callus, passes through screening and optimizing Fiber differentiation
Base and subculture medium, acquisition be suitable for the nutritional ingredient of long-term preservation is carried out under low temperature or ultralow temperature, while by low temperature or
After the callus of cryopreservation is inoculated into recovery subculture medium, can quickly restore normal growth state, be broken up
And grow up to test tube seedling.By the application to save callus required for Fiber differentiation, save subculture medium and recovery after
It for each component type of culture medium and its optimization of content, can significantly extend storage life, reduce the number of regular subculture, greatly
It is big to reduce manual labor amount, moreover it is possible to reduce the probability that multiple subculture causes varied in ploidy, and also not influence callus just
Often differentiation, growth, are conducive to provide good callus at any time, to produce a large amount of benign systems, monoploid, polyploid or work
Sufficient source is provided for scientific research material.
Further advantage, target and feature of the invention will be partially reflected by the following instructions, and part will also be by this
The research and practice of invention and be understood by the person skilled in the art.
Detailed description of the invention
Fig. 1 is that the present invention is stored at room temperature 2 months carrot callus of rear renewal cultivation；
The carrot callus that Fig. 2 is renewal cultivation 2 months after deepfreeze of the present invention saves；
The carrot callus that Fig. 3 is renewal cultivation 2 months after cryopreservation of the present invention；
Fig. 4 is that the carrot after cryopreservation recovery subculture of the present invention breaks up seedling.
Specific embodiment
The present invention is described in further detail below, to enable those skilled in the art's refer to the instruction text being capable of evidence
To implement.
It should be appreciated that such as " having ", "comprising" and " comprising " term used herein are not precluded one or more
The presence or addition of a other elements or combinations thereof.
Embodiment 1
Extend the medium component of carrot callus' storage life and the screening of temperature condition
Because Plant Tissue Breeding is relatively time-consuming and consumptive material, in order to save time with corresponding cost, the preservation of callus
It is necessary.Different nutrients is most important to the growth of plant callus, and therefore, the present invention is protecting
Before depositing, the squamous subculture based component saved to carrot callus is optimized, in routine preservation subculture medium (MS+
0.5mg/L NAA+0.3mg/L 2,4-D+7.0g/L agar) on the basis of adjust sucrose, mannitol and abscisic acid three kinds of substances
Content, by Orthogonal Experiment and Design at 9 formulas (being shown in Table 1), each each solution for preparing 200mL of formula.Each experimental material
It is repeated twice, the culture medium of every cuvette cartridge 20mL, respectively in room temperature (25 DEG C), (4 DEG C) of low temperature refrigerations and (- 20 DEG C of ultralow temperature
With -80 DEG C) freezing under cultivated, observe the growing state of each experimental material.
The callus of 1 orthogonal of table saves the composition of additive and corresponding amount in formula
(note: 1% indicates to add 10g sucrose or mannitol in every 1L MS culture medium；2% and 3% and so on)
The growing state of the culture medium that 2 orthogonal experiment of table is accordingly formulated at different temperatures
(note: growing state is the result for being inoculated with 1 month (30 days) and (60 days) 2 months statistics in table)
From Table 2, it can be seen that saving carrot callus under normal temperature, formula 3 and 6 is inhibiting growth result ratio
Preferably；With normal temperature phase ratio, low temperature (4 DEG C) general trend is all to grow more slow, and freezing conditions almost lose growth, due to
Icing is unfavorable for observing one by one, so being not described in detail result.
In conjunction with the growing state of room temperature and low temperature totality, low temperature and room temperature are in induced medium MS+0.2mg/L KT+
Inhibit life when addition sucrose 1-2%, mannitol 3% on the basis of 0.2mg/L 2,4-D+7.0g/L agar, 50 μm of ol/L of ABA
Long effect is best.Whether and above-mentioned substance addition can achieve the mesh for extending callus preservation under the conditions of freezen protective or not
's.It is observed that the material under normal temperature, which can be reserved for, does not have to subculture for 1 year or so, low temperature or refrigeration material can be reserved for long to 3 to 5
Year in addition it is longer.
Embodiment 2
1, the formula of test reagent used in the embodiment of the present invention is as follows:
MS culture medium mother liquor is prepared: the preparation routinely MS training of a great number of elements, microelement, molysite and organic substance mother liquor
Based formulas is supported, a large amount of to expand 10 times, remaining expands 100 times and prepares corresponding mother liquor, saves in 4 DEG C of refrigerators.
The preparation of auxin mother liquor: two kinds of auxin of NAA (methyl α-naphthyl acetate) and 2,4-D (dichlorphenoxyacetic acid) use 1M respectively
NaOH or the dissolution of 95% alcohol, use ultrapure water constant volume afterwards, make its concentration 1mg/mL, the mother liquor prepared saves in 4 DEG C of refrigerators.
The preparation of the mother liquor of the basic element of cell division: a small amount of 1M HCL is added in 6-BA (6-benzyl aminopurine), KT (kinetin)
Dissolution finally uses ultrapure water constant volume, makes its concentration 1mg mL-1, the mother liquor prepared saves in 4 DEG C of refrigerators.
Induced medium is containing 0.1mg/L KT, 2 0.2mg/L, the MS training of 4-D, 25g/L sucrose and 7.0g/L agar
Support base.
Saving subculture medium is to contain 0.1mg/L KT, 2 0.1mg/L, 4-D, 20g/L sucrose, 30g/L mannitol, 50
The MS culture medium of μm ol/L ABA and 7.0g/L agar.
Recovery subculture medium used in the carrot callus of cryo-conservation are as follows: contain 0.2mg/L KT, 0.2mg/L
2,4-D and sucrose 25g/L and 7.0g/L agar MS culture medium；
Liquid rejuvenation culture medium used in the carrot callus of cryopreservation presses body by fluid nutrient medium and glycerol
Product is mixed than 0.4-1:1, and the fluid nutrient medium is containing 0.2mg/L KT, 1.0mg/L NAA and sucrose 25g/L
MS culture medium；Solid rejuvenation culture medium used is to contain 0.2mg/L KT, 1.0mg/L NAA, sucrose 25g/L and agar
The MS culture medium of 7.0g/L.
2, extend the method for the method and carrot callus' recovery regrowth of carrot callus' storage life
(1) it chooses after the carrot in current year raw harvesting two weeks cleans, impregnates 30s with 75% alcoholic solution, then with 0.1%
Mercuric chloride solution impregnates 15min, uses aseptic water washing 4 times later, is placed on aseptic paper and absorbs water；Then stripping and slicing, then by stripping and slicing
Carrot explant be inoculated in induced medium, in 25 DEG C of dark condition cultures to growing callus；The Fiber differentiation
Base is to contain 0.1mg/L KT, 2 0.2mg/L, the MS culture medium of 4-D and 25g sucrose and 7.0g/L agar；
(2) callus grown is inoculated in and is saved in subculture medium, be placed in 4 DEG C of low temperature or condition of ultralow temperature
It is saved under (- 20 DEG C or -80 DEG C), the preservation subculture medium is to contain 0.1mg/L KT, 2 0.1mg/L, 4-D, 20g sugarcane
The MS culture medium of sugar, 30g mannitol and 50 μm of ol/L ABA, wherein need to save described after being commissioned to train when 4 DEG C of cryo-conservations
Agar is added by 7.0g/L on the basis of feeding base,；It saves under the conditions of -20 DEG C of low temperature or -80 DEG C of superfreeze without addition).
(3) carrot callus of cryo-conservation are inoculated on recovery subculture medium, observation period growth；Recovery subculture
Culture medium is to contain 0.2mg/L KT, 2 0.2mg/L, the MS culture medium of 4-D and sucrose 25g/L and agar 7.0g/L；
The carrot callus of cryopreservation be first transferred in liquid rejuvenation culture medium (fluid nutrient medium be containing
The MS culture medium of 0.2mg/L KT, 1.0mg/L NAA and sucrose 25g/L after fluid nutrient medium high pressure sterilization, in addition prepare matter
Glycerol and the sterilizing that score is 50%, 80% are measured, then match according to 3:7,4:6,5:5 by glycerol and fluid nutrient medium
To the mixed liquor of glycerol and culture medium to get the liquid rejuvenation culture medium), each formula is repeated twice.According to zooblast
The principle of recovery then after super-clean bench completes switching, is first put into 4 DEG C of refrigerator half a day, next puts on the basis of melting fastly
Enter -20 DEG C of refrigerator overnights, finally place into -80 DEG C of refrigerator overnights, is sequentially placed after taking out each in 30 DEG C and 35 DEG C of water-baths
It after half an hour, brings super-clean bench into and carries out fluid nutrient medium culture after a week, go to (the solid rejuvenation training of solid rejuvenation subculture medium
Supporting base is the MS culture medium containing 0.2mg/L KT, 1.0mg/L NAA, sucrose 25g/L and agar 7.0g/L), observation growth
Situation.As a result, it has been found that the carrot callus that ultralow temperature (freezing) saves are transferred to liquid rejuvenation after being commissioned to train through water-bath thawing
Support after recovering on base, find liquid medium within: glycerol is in the mixed culture medium of 3:7, whether containing 50% glycerol or
All there is brown stain in 80% glycerol, the growth of remaining 4:6 or 5:5 are all fine.Therefore, the carrot callus of freezen protective are multiple
The glycerol of addition 40% or 50%, is conducive to the restoration ecosystem of callus in the fluid nutrient medium of Soviet Union.
3, the recovery situation under condition of different temperatures
3.1 carrot callus saved at (25 DEG C) of normal temperature condition, change the type and content of hormone, and observation is different
Recovery subculture medium in carrot callus' increment variation；As shown in table 3, in 9 kinds of recovery subculture mediums, In
The increment of formula 4 and formula 5 is maximum after 2 months, shows that squamous subculture based formulas is MS+25g/L+1.0mg/L KT+0.1-
0.2mg/L 2,4-D+ agar 7.0g/L；The state of the carrot callus of room temperature culture 2 months is as shown in Figure 1.
Table 3 adds the variation of callus incrementss in hormon and its recovery subculture medium of content
(callus being stored at room temperature)
3.2 cryo-conservation carrot callus, change the type and content of hormone, observe different recovery subcultures
The variation of carrot callus' increment in culture medium；As shown in table 4, in 9 kinds of recovery subculture mediums, 1 is formulated after 2 months
It is maximum with the increment of formula 2, show that squamous subculture based formulas is MS+25g/L sucrose+0.2mg/L KT+0.2-0.5mg/L2,
4-D+ agar 7.0g/L；Squamous subculture 2 months carrot callus after the cryo-conservation recovery that Fig. 2 is shown.
Table 4 adds the variation of callus incrementss in hormon and its recovery subculture medium of content
(callus of cryo-conservation)
3.3 freezen protective carrot callus, change the type and ratio of hormone, observe different recovery subcultures
The variation of carrot callus' increment in culture medium；As shown in table 5, in 9 kinds of recovery subculture mediums, 3 are formulated after 2 months
It is maximum with the increment of formula 6, show that recovery squamous subculture based formulas is MS+25g/L sucrose+0.2-0.5mg/L KT+
(this is liquid resuscitation subculture medium to 1.0mg/L NAA, and solid recovery subculture medium adds 7.0g/L fine jade on this basis
Rouge)；Squamous subculture 2 months carrot callus after the Cryopreservation that Fig. 3 is shown.
Table 5 adds the variation of callus incrementss in hormon and its recovery subculture medium of content
(callus of cryo-conservation)
Embodiment 3
The carrot callus saved respectively under 3.1 room temperature, low temperature and the condition of ultralow temperature obtained according to embodiment 2,
The culture medium that the optimum obtained after optimization carries out recovery squamous subculture carries out recovery growth respectively, and culture calculates separately after 30 days
The relative survival of callus under different temperatures, the results are shown in Table 6, it will be apparent that, it recovers again after low temperature and cryopreservation
The relative survival of growth is higher than the relative survival for the callus being stored at room temperature, it may be possible to be easy since normal temperature condition saves
Cause nutriment exhaustion, the accumulation of moisture loss and metabolite, causes the appearance of partial differentiation proliferation phenomenon, be unfavorable for
Long-term preservation, and cryo-conservation and cryopreservation effectively inhibit differentiation and proliferation, and callus is remained at undifferentiated increasing
State is grown, the nutritional status required for suitable growth and differentiation always is allowed to, to be conducive to promote recovery and raising recovery
The relative survival of regrowth.
Above-mentioned experimental material be respectively adopted room temperature (25 DEG C, control), low temperature (4 DEG C), (- 80 DEG C) of ultralow temperature save 2 months
Carrot callus.
Recovery situation after 6 different temperatures of table saves
(note: after compareing to be stored at room temperature, the relative survival of room temperature culture；Test 1 is room temperature culture after cryo-conservation
Relative survival；Test 2 is the relative survival of room temperature culture after cryopreservation)
3.2 according to 3.1 training method, the carrot callus of cryopreservation are transformed into recovery subculture medium
After 2 months, then it is transformed into differential medium MS+25g/L sucrose+7.0mg/L KT+0.2mg/L 2,4-D+ agar 7.0g/L, In
25 DEG C, the case where irradiating 6-8h under 1200Lux illumination daily, observe restoration ecosystem, as shown in figure 4, to be transferred to differentiation training
Test tube seedling when 3 months after feeding base, illustrates that the carrot callus of cryopreservation being capable of normal restoration ecosystem.
Although the embodiments of the present invention have been disclosed as above, but its is not only in the description and the implementation listed
With it can be fully applied to various fields suitable for the present invention, for those skilled in the art, can be easily
Realize other modification, therefore without departing from the general concept defined in the claims and the equivalent scope, the present invention is simultaneously unlimited
In specific details and shown here as with attached drawing.
Claims (7)
1. a kind of method for extending carrot callus' storage life, which comprises the following steps:
Step 1: after choosing current year raw carrot root tuber pretreatment, stripping and slicing, then the carrot explant of stripping and slicing is inoculated in induction
Culture medium, in dark condition culture to growing callus；The induced medium is to contain 0.1-0.2mg/L KT, 0.2mg/
The MS culture medium of L 2,4-D and 25-30g/L sucrose and 6.5-7.5g/L agar；
Step 2: the callus grown being inoculated in and is saved in subculture medium, is placed under low temperature or condition of ultralow temperature and protects
It deposits, the preservation subculture medium is to contain 0.1-0.2mg/L KT, 2 0.1-0.2mg/L, 4-D, 10-20g/L sucrose, 25-
The MS culture medium of 30g/L mannitol and 50 μm of ol/L ABA, wherein when cryo-conservation, in the preservation subculture medium
On the basis of by 6.5-7.5g/L add agar.
2. extending the method for carrot callus' storage life as described in claim 1, which is characterized in that selected in step 1
Carrot be without wound, healthy growth uniformly and harvesting 2 weeks within fresh carrot root tuber, protected under the conditions of 4 DEG C after harvesting
Storage hiding；
Carrot root tuber preprocess method are as follows: will choose carrot clean after, with mass fraction be 75% alcoholic solution impregnate 30-
40s, then with mass fraction be 0.1%-0.2% mercuric chloride solution impregnate 10-15min, later use aseptic water washing 4-6 times, be placed in
Suck dry moisture on aseptic paper.
3. extending the method for carrot callus' storage life as described in claim 1, which is characterized in that cultivated in step 1
Temperature is 25-27 DEG C.
4. extending the method for carrot callus' storage life as described in claim 1, which is characterized in that low temperature is 4 DEG C；It is super
Low temperature is -20 DEG C or -80 DEG C.
5. a kind of method of carrot callus' recovery regrowth, which is characterized in that the carrot callus are to use
The carrot callus that the described in any item methods of claim 1-4 save, carrot callus' recovery regrowth
Method specifically: the carrot callus that will be saved under low temperature or condition of ultralow temperature, be inoculated in respectively different recoveries after
For on culture medium, cultivated under the conditions of 25-27 DEG C.
6. the method for carrot callus' recovery regrowth as claimed in claim 5, which is characterized in that the Hu of cryo-conservation
The recovery subculture medium of radish callus be containing 0.2mg/L KT, 2 0.2-0.5mg/L, 4-D, sucrose 20-30g/L and
The MS culture medium of agar 6.5-7.5g/L；
The carrot callus of cryopreservation are first transferred in liquid rejuvenation culture medium after defrosting according to method is melted fastly, then
It is transferred in solid rejuvenation culture medium and cultivates, restoration ecosystem；The liquid rejuvenation culture medium be by fluid nutrient medium and glycerol by
Volume ratio 0.4-1:1 is mixed, and the fluid nutrient medium is to contain 0.2-0.5mg/L KT, 1.0mg/L NAA and sucrose
The MS culture medium of 20-30g/L；The solid rejuvenation culture medium is to contain 0.2-0.5mg/L KT, 1.0mg/L NAA, sucrose 20-
The MS culture medium of 30g/L and agar 6.5-7.5g/L.
7. the method for carrot callus' recovery regrowth as claimed in claim 6, which is characterized in that the side of melting fastly
Method, specifically: the callus being inoculated into the liquid rejuvenation culture medium is first put into 4 DEG C of refrigerators 6 hours, places into -20 DEG C
Refrigerator overnight is put into -80 DEG C of refrigerator long-term preservations later, after needing to take out when recovery, is sequentially placed in 30 DEG C and 35 DEG C of water-baths
Pot each half an hour completes to thaw.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111387058A (en) * | 2020-05-12 | 2020-07-10 | 中国农业科学院蔬菜花卉研究所 | Ultralow-temperature preservation method and ultralow-temperature preservation equipment for garlic callus |
| CN115380823A (en) * | 2022-08-29 | 2022-11-25 | 安徽农业大学 | Method for delaying subculture growth rate of tissue culture seedlings of black-bone vegetables |
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