CN107410024A - A kind of abductive approach of avocado callus and the method for promoting its bud to break up - Google Patents
A kind of abductive approach of avocado callus and the method for promoting its bud to break up Download PDFInfo
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- CN107410024A CN107410024A CN201710345217.XA CN201710345217A CN107410024A CN 107410024 A CN107410024 A CN 107410024A CN 201710345217 A CN201710345217 A CN 201710345217A CN 107410024 A CN107410024 A CN 107410024A
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- avocado
- callus
- explant
- abductive approach
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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 provides a kind of abductive approach of avocado callus and the method for promoting its hair to break up.The abductive approach of the avocado callus, comprises the following steps：Avocado explant is inoculated into inducing culture, cultivated 4~30 days under conditions of 1200~1700Lx of intensity of illumination, 8~14h/ days photoperiod, 22~28 DEG C of temperature, obtain callus, wherein, the MS basal mediums that it is 5.8~6.5 containing 20~40g/L sucrose, 5~12g/L agar, 0~3.0mg/L methyl α-naphthyl acetates, the dichlorphenoxyacetic acids of 0.1~3.0mg/L 2,4, the benzyl aminoadenines of 0.1~3.0mg/L 6 and pH that the inducing culture, which is,.The abductive approach of avocado callus provided by the present invention is high using the benzyl aminoadenine of low concentration 6 and 2, the combination of 4 dichlorphenoxyacetic acid hormonal components, the inductivity of the avocado callus cultivated；The present invention is studied explant types such as blade, stem section, bud sections, it is found that the inductivity of stem section explant is up to more than 90%, for further successfully induction intact plant lays the foundation from now on.
Description
Technical field
The present invention relates to technical field of plant propagation, and in particular to a kind of abductive approach of avocado callus and promotion
The method of its bud differentiation.
Background technology
Avocado (Persea Americana Mill) belongs to the evergreen orchard fruit of Lauraceae Persea, and nutritive value enriches, and
With certain medicinal and health-care effect.Avocado tradition modes of reproduction breeding coefficient is low, and pest and disease damage etc. limits its plantation and development；One
Determine can solve this problem by tissue culture technique in degree.Tissue culture technique have profitable, modes of reproduction is more, culture
The advantages that cycle is short, this technology are studied and applied in fruit such as banana, papaya and strawberries.
Abroad, (zygote formed with Gamete Fusion by the male and female of plant is then with immature zygotic embryo by most scholars
Develop the sexual embryo formed) it is explant；Body embryo is induced by direct/indirect body embryogenesis path and realizes avocado plant again
It is raw.Carlos L ó pez Encina et al. with " Reed ", " Hass ", " Duke 7 ", " A10 " avocado kind prematurity zygote
Embryo is explant, inquires into the influence of different cultural methods and different cultivars to somatic embryo regeneration plant induction.As a result show ' Reed '
Avocado (Persea Americana Mill.cv.Reed) body embryo incidence is higher, and by two step cultivations (solid state rheology-
Liquid culture-solid state rheology) and add in the medium 0.4g/L proline or 1g/L glutamine be advantageous to body embryo into
Ripe and sprouting；But the probability of body embryo the two poles of the earth growth is still relatively low (bud and root grow simultaneously).Rohim inquires into embryo age, culture base class
Type and sucrose concentration find that embryo age is that 180d, MS are minimal medium and sucrose concentration is to the influence that body embryo is ripe and sprouts
During 30g/L, its best results.But it is that explant induces avocado by direct or indirect adventitious organogenesis with blade, stem section etc.
The correlative study of intact plant is less.At present, only see that Young is studied using " Lula " kind avocado blade, stem section as explant
The influence of single hormone kind, temperature and illumination to callus induction；But the differentiation of callus bud is not ground further
Study carefully.Barrera-Guerra et al. induces avocado intact plant using avocado stem section as explant by direct adventitious organogenesis,
Inquire into the influence of growth hormone species and its concentration to culture of rootage.It was found that indirect rooting method effect is preferable, but rooting rate is still
It is relatively low.
In China, the research on avocado tissue cultures is less, at present only Liang Gao, Peng Minzhang and He Bizhu et al. couple
It carried out the research of correlation.Liang Gao and He Bizhu et al. pass through direct organ using avocado bearing tree new life stem section as explant
A development ways induction Multiple Buds step of going forward side by side realizes avocado plant regeneration.Peng Mingzhang is using hypocotyl as explant evoked callus
And combine cryogenic freezing Techniques of preserving and inquire into the correlative factor for influenceing preserving seed.
At present, not yet reported research hormon species and its concentration and explant type to " Haas " avocado callus group
The influence of induction and bud differentiation is knitted, to filter out optimal inductive condition and explant.
The content of the invention
In order to overcome above-mentioned deficiency, the invention provides a kind of abductive approach of avocado callus and promote its bud point
The method of change.
The technical proposal for solving the technical problem of the invention is：
In a first aspect, the invention provides a kind of abductive approach of avocado callus, comprise the following steps：
Avocado explant is inoculated into inducing culture, in 1200~1700Lx of intensity of illumination, 8~14h/ of photoperiod
My god, cultivate 4~30 days under conditions of 22~28 DEG C of temperature, obtain callus, wherein, the inducing culture be containing 20~
40g/L sucrose, 5~12g/L agar, 0~3.0mg/L methyl α-naphthyl acetates, 0.1~3.0mg/L 2,4 dichlorophenoxyacetic acids, 0.1~
The MS basal mediums that 3.0mg/L 6- benzyls aminoadenines and pH are 5.8~6.5.
Preferably, the avocado explant include but is not limited to avocado blade, avocado petiole, avocado stem apex, avocado stem section,
One or more in avocado bud section.
It is further preferred that the avocado explant is avocado stem section.
Preferably, the kind of the avocado include but is not limited to " Haas " avocado, " Reed " avocado, " avocados of Duke 7 ",
One kind in " A10 " avocado.
It is further preferred that the kind of the avocado is " Haas " avocado.
Preferably, the cultivation time of the avocado callus is 6~14 days.
Preferably, the inducing culture be containing 20~40g/L sucrose, 5~12g/L agar, 0.1~3.0mg/L 2,
4- dichlorphenoxyacetic acids, 0.1~3.0mg/L6- benzyls aminoadenine and pH are 5.8~6.5 MS basal mediums.
It is further preferred that the inducing culture be containing 20~40g/L sucrose, 5~12g/L agar, 0.1~
2.0mg/L 6- benzyls aminoadenine, 0.1mg/L 2,4 dichlorophenoxyacetic acids and pH are 5.8~6.5 MS basal mediums.
It is further preferred that the inducing culture be containing 20~40g/L sucrose, 5~12g/L agar, 0.1~
2.0mg/L6- benzyls aminoadenine, 0.1mg/L 2,4 dichlorophenoxyacetic acids and pH are 5.8~6.5 MS basal mediums.
It is further preferred that the inducing culture is to contain 30g/L sucrose, 7g/L agar, 0.5mg/L 6- benzyl ammonia
Base adenine, 1.0mg/L 2,4 dichlorophenoxyacetic acids and pH are 5.8~6.5 MS basal mediums.
Preferably, the explant is that the avocado sample sterilization collected is made.
It is further preferred that the explant is to be made using following methods：
The avocado sample of collection, it is put on superclean bench, adds alcohol and carry out surface sterilization about 15~25s, with nothing
Bacterium water cleans 2~3 times, is transferred to HgCl2Sterilize about 4~5min in solution, then with aseptic water washing 4~5 times；By the oil after processing
Pears stem section is put on aseptic filter paper after suck dry moisture, and explant is made；Wherein, the concentration of the alcohol is not less than 70%；It is described
HgCl2The concentration of solution is 0.05~1%.
Preferably, in the abductive approach of the avocado callus, the inductivity of the avocado callus is not less than
75%.
Second aspect, present invention also offers a kind of method for promoting the differentiation of avocado callus bud, comprise the following steps：
The callus for taking the abductive approach of avocado callus as described in relation to the first aspect to cultivate, by the callus group of gained
Knit to be inoculated into bud differential medium and cultivate 30~50 days, the bud differential medium be containing 20~40g/L sucrose, 5~
12g/L agar, 0.1~3.0mg/L methyl α-naphthyl acetates, 0.1~3.0mg/L 6- benzyls aminoadenines and pH are 5.8~6.5 MS bases
Basal culture medium.
Preferably, in the method for promoting avocado callus bud to break up, the bud ratio of the avocado callus is not
Less than 35~60%.
By adopting the above-described technical solution, the present invention has the advantages that compared with prior art：
In the prior art, carry out evoked callus through being combined frequently with a variety of hormonal components, or use high-concentration naphthalene second
The combination of acid, 2,4- dichlorphenoxyacetic acids or two kinds of hormones carrys out evoked callus, and avocado callus group provided by the present invention
The abductive approach knitted, combined, cultivated using low concentration 6- benzyls aminoadenine and 2,4- dichlorphenoxyacetic acid hormonal components
Avocado callus inductivity it is high；The present invention is studied explant types such as blade, stem section, bud sections, finds stem
The inductivity of section explant is up to more than 90%, and bud ratio is high, for further successfully induction intact plant lays the foundation from now on.
Brief description of the drawings
Fig. 1 is the callus group of " Haas " avocado blade induced synthesis under the various concentrations NAA that the embodiment of the present invention is provided
The dry weight and fresh weight of plant seedlings knitted；
Fig. 2 is the callus of " Haas " avocado blade induced synthesis under the various concentrations 2,4-D that the embodiment of the present invention is provided
Organize dry weight and fresh weight of plant seedlings；
Fig. 3 is the callus group of " Haas " avocado blade induced synthesis under the various concentrations 6-BA that the embodiment of the present invention is provided
Knit dry weight and fresh weight of plant seedlings；
The callus for " Haas " avocado different parts explant induced synthesis that Fig. 4 embodiment of the present invention is provided is fresh and dried
Weight；
Fig. 5 is the growing state that the embodiment of the present invention provides " Haas " avocado callus.
Embodiment
Below by embodiment, the present invention is described in detail.
It is understood that the embodiment of the present invention is arranged to data, calculated using Excell softwares, average value is used
The integral level of each processing is represented, the difference of each parameter is analyzed using the ANOVA of SPSS19.0 statistical softwares.Callus induction
The calculation formula of rate, water content and moisture content is as follows：
Callus induction rate=(the explant sum for forming explant number/inoculation of callus) * 100%；
Water content (g)=fresh weight-dry weight；
Callus moisture content=(callus water content/callus fresh weight) * 100%；
Bud ratio=(budding callus block number/inoculation callus block number) * 100%.
The influence that the different single hormone kinds of embodiment 1 and its concentration are induced " Haas " avocado Callus of Leaf
The embodiments of the invention provide a kind of abductive approach of " Haas " avocado callus, comprise the following steps：
(1) explant sterilization：" Haas " avocado blade of collection, it is put on superclean bench, adds 75%
Alcohol carries out surface sterilization about 15~25s, with sterile water wash 2~3 times, is transferred to 0.1%HgCl2Sterilize about 4 in solution~
5min, then with aseptic water washing 4~5 times；Explant is put on aseptic filter paper after suck dry moisture, under the conditions of gnotobasis,
Blade afterbody and marginal portion are cut, stays close to master pulse part, is cut into 5*5mm sizes；Explant is made, it is standby；
(2) callus tissue culture：Explant obtained by step (1) is inoculated into addition respectively containing various concentrations
(0.2,0.4,0.6,0.8,1.0,2.0,3.0mg/L) methyl α-naphthyl acetate (NAA) and (0.2,0.4,0.6,0.8,1.0,2.0,3.0mg/
L) in the inducing culture of 2,4- dichlorphenoxyacetic acids (2,4-D), wherein, the inducing culture with MS basal mediums,
Also include 30g/L sucrose, 7g/L agar, pH is 6.0 ± 0.1.Often processing is inoculated with 30 explants or so, is repeated 3 times, every bottle connects
2~3 explants of kind.Callus is obtained after cultivating 30 days.Wherein, condition of culture is：Intensity of illumination 1700Lx, photoperiod
Cultivated in the tissue culture room of 14h/ days (daily 10h is dark), 25 ± 2 DEG C of temperature；Routine observation, count callus induction
Rate, dry weight and fresh weight of plant seedlings, screen most suitable callus inducing medium.
Analysis of experimental results：
(1) routine observation result：Blade explant is inoculated into the NAA containing various concentrations and 2,4-D Fiber differentiation
In base, it is inoculated with 2 weeks or sos, in incision appearance expanding and form macroscopic callus in various degree.Main distribution
At explant otch both ends, minority is scattered to be distributed in explant surface, and small volume, milky, quality is harder, and particle is presented more
Spherical shape (see Fig. 5 A, B).If incubation time is long, the easy browning of callus of formation is dead.
(2) hormon species and its concentration to the influence that " Haas " avocado Callus of Leaf induces referring to table 1 and
Fig. 1 and Fig. 2.
As shown in Table 1, the influence of variety classes hormone and its concentration to callus induction is notable.Without any hormone
Culture medium in, there is not any change in explant.And in the NAA containing various concentrations and 2,4-D culture medium, part
Can induced synthesis callus.Its inducing effect is preferable during addition 2,4-D, and inductivity is 10.6~45.3%, when 2,4-D concentration
For 1.0mg/L when its inductivity be up to 45.3%.Dropped afterwards as the increase callus induction rate of 2,4-D concentration first increases
It is low, illustrate that 2,4-D of high concentration is unfavorable for the induction of callus.In the medium add NAA when, its inductivity be 5.7~
20.2%；When NAA concentration is 1.0mg/L, its inductivity is up to 20.2%.With the increase of NAA concentration, its induction takes the lead in
Reduced after increase, illustrate that high concentration NAA is unfavorable for the induction of callus.
Find out the increase with hormone concentration from Fig. 1 and 2, the fresh weight and dry weight of the callus of formation first increase to drop afterwards
It is low.When 2,4-D and NAA concentration are 1.0mg/L, its fresh weight reaches maximum, respectively 0.83g and 0.37g.
Generally speaking, when adding single kind of growth hormone, " Haas " avocado Callus of Leaf inducing effect is not to manage very much
Think, may coordinate the basic element of cell division using when its effect can make moderate progress, this needs further research and inquirement.
The influence that the hormon species of table 1 and its concentration are induced " Haas " avocado Callus of Leaf
The influence that the different hormone combinations of embodiment 2 are induced " Haas " avocado Callus of Leaf
In order to further illustrate beneficial effects of the present invention, the step of repeating the embodiment of the present invention 1, by the embodiment of the present invention
Hormonal components in 1 the step of (2) in inducing culture are substituted for hormonal components as shown in table 2.
Analysis of experimental results：
(1) routine observation result：Blade is inoculated into respectively in 6 kinds of culture mediums, inoculation 6d or so, explant starts swollen
Greatly, the callus of yellowish white is formed in cutting part dedifferentiation.As incubation time extends, the callus of formation is gradual
Expand, slowly cover explant；At the latest form callus in inoculation 14d or so.The callus of formation is loose, light yellowish-white
Color, water content are more；The callus that part is formed does not form callus group along with flocculent white material (see figure C), part
The explant knitted expands in incision and browning is presented.Inoculation 30d or so starts to count and transferred, it is found that incubation time is long more
The easy browning of injured tissue.
(2) as shown in Table 2,0.1~2.0mg/L 6-BA, 1.0mg/L 2,4-D and 0.1mg/L are added in the medium
NAA can induce callus, and effect is best when 6-BA concentration is 0.5mg/L, and its inductivity is 55.8%；In MS+
Inducing effect in 0.5mg/L 6-BA+1.0mg/L 2,4-D+0.1mg/LNAA culture mediums is with MS+0.5mg/L 6-BA+
The inducing effect of 1.0mg/L 2,4-D culture mediums is suitable；And in MS+0.1mg/L 6-BA+1.0mg/L 2,4-D culture mediums more
Injured tissue inducing effect is poor, and the callus browning of induced synthesis is more serious.
As can be seen from Figure 3 with the increase of 6-BA concentration, the fresh weight of the callus of formation first increases to be reduced afterwards, explanation
High concentration 6-BA is unfavorable for the induction of callus.The moisture content of callus is 93~95.9%, when 6-BA concentration is
Its fresh weight is maximum during 0.5mg/L, is 3.82g, and is cultivated in MS+0.5mg/L 6-BA+1.0mg/L 2,4-D+0.1mg/LNAA
The fresh weight of the callus of induced synthesis is 0.315g in base, and both differences are little；Illustrate in MS+0.5mg/L 6-BA+
It is not apparent that 1.0mg/L 0.1mg/L NAA effects are added in 2,4-D culture mediums.
The influence that the different culture media of table 2 is induced " Haas " avocado Callus of Leaf
Influence of the different explant types of embodiment 3 to " Haas " avocado callus induction
In order to further illustrate beneficial effects of the present invention, the step of repeating the embodiment of the present invention 1, by the embodiment of the present invention
" avocado blade " is substituted for explant as shown in table 3 in 1 the step of (1).
(1) routine observation result：Avocado blade, petiole, stem apex, stem section (band/without effective bud point) are inoculated into MS+
In 0.5mg/L 6-BA+1.0mg/L 2,4-D minimal medium, time of callus induction, quality, color etc. are observed.Connect
Kind 4d or so explant starts to expand, and 7d or so can see macroscopic callus；The callus milky of formation,
It is loose, the callus that more (see Fig. 5 D-K) the part explant of water content is formed with white flock material (see Fig. 5 F, G, H,
K)
(2) as shown in Table 3, blade, petiole, stem apex, stem section (band/without effective bud point) can evoked callus, and
Influence of the explant type to callus induction is notable.The inducing effect of stem section is optimal, without the induction of effective bud point stem section
Rate is 98.3%, and the inductivity of the stem section with effective bud point is 72.7%；It is probably the callus group formed with effective bud point stem section
It is sporadicly to be distributed in explant surface to knit mostly, and is accompanied by except forming callus in addition to the elongation of axillary bud and grows therefore to disappear
Part nutrition is consumed.5 kinds of explant type inducing effects it is worst be stem apex, its inductivity be 20%, it may be possible to culture medium and
Condition of culture is unfavorable for its induced synthesis callus.
Fig. 4 understands to be existed by the callus mean fresh of blade, petiole, stem apex, stem section (band/without effective bud point) induced synthesis
2.25~3.415g, average dry weight is in 0.13~0.15g, and moisture content is 93.9~96.2%.When explant is without effective bud
Its fresh weight is maximum during point stem section, is 3.415g.
Influence of the different parts explant of table 4 to " Haas " avocado callus induction
Influence of the different callus tissue culture times of embodiment 4 to bud ratio
The embodiments of the invention provide a kind of method for improving avocado callus induction and growth, comprise the following steps：
(1) explant sterilization：" Haas " the avocado stem section (without effective bud point) of collection, superclean bench is put into
On, the alcohol for adding 75% carries out surface sterilization about 15~25s, with sterile water wash 2~3 times, is transferred to 0.1%HgCl2Solution
Middle sterilizing about 4~5min, then with aseptic water washing 4~5 times；Avocado stem section after processing is put into suck dry moisture on aseptic filter paper
Afterwards, explant is made, it is standby；
(2) callus tissue culture：Explant obtained by step (1) is inoculated into inducing culture respectively, wherein, it is described
Inducing culture is to contain 30g/L sucrose, 7g/L agar, 0.5mg/L methyl α-naphthyl acetates (NAA), 1.0mg/L 2,4 dichloro benzene oxygen second
The MS basal mediums that sour (2,4-D) and pH are 6.0 ± 0.1；Often processing is inoculated with 30 explants or so, is repeated 3 times, every bottle connects
2~3 explants of kind.Callus is obtained after cultivating 30 days.Wherein, condition of culture is：The illumination of intensity of illumination 1500Lx, 12h/
12h is dark, and temperature is to carry out dark culturing in 25 ± 2 DEG C of tissue culture room；Routine observation, statistics callus induction rate, fresh and dried
Weight, cultivate 5,7,10,14,18,21,30 days obtain callus respectively；
(3) bud differentiation culture：Callus obtained by step (2) is inoculated into bud differential medium respectively, the bud
Differential medium is to contain 30g/L sucrose, 7g/L agar, 0.5mg/L methyl α-naphthyl acetates (NAA), 1.0mg/L 6- benzyl aminoadenines
(6-BA) and pH are 6.0 ± 0.1 MS basal mediums；30 explants of often processing inoculation, are repeated 3 times, observe growing state
And in 40 days or so statistical results.
Analysis of experimental results：As shown in table 5, the callus obtained after cultivating 30 days carries out bud differentiation culture and browning occurs
It is dead with different degrees of hair, it is difficult to differentiation budding, and cultivate 6,10,14,18 days callus obtained, do not occur browning or
Browning degree is small, and bud ratio is far above the callus obtained after cultivating 21,30 days.
Influence of the different callus tissue culture times of table 5 to bud ratio
Influence of the 5 same inducing culture of embodiment to the callus induction of the explant of different cultivars
(1) explant sterilization：Gather the stem section of following kind fruit tree respectively (without effective bud point)：" Haas " oil
Pears, avocado, " Lula " avocado, " avocados of Duke 7 ", " Reed " avocado, are put on superclean bench, the alcohol for adding 75% enters
Row surface sterilization about 15~25s, with sterile water wash 2~3 times, it is transferred to 0.1%HgCl2Sterilize about 4~5min in solution, then uses
Aseptic water washing 4~5 times；Explant is put on aseptic filter paper after suck dry moisture, under the conditions of gnotobasis, blade afterbody
Cut with marginal portion, stay close to master pulse part, be cut into 5*5mm sizes；" Haas " avocado blade explant, avocado leaf is made
Piece, " Lula " avocado blade explant, " the avocado blades of Duke 7 " explant, " Reed " avocado blade explant, it is standby；
(2) callus tissue culture：Explant obtained by step (1) is inoculated into respectively in addition inducing culture, wherein,
The inducing culture is to contain 30g/L sucrose, 7g/L agar, 0.5mg/L methyl α-naphthyl acetates (NAA), 1.0mg/L 2,4 dichloro benzenes
The MS basal mediums that fluoroacetic acid (2,4-D) and pH are 6.0 ± 0.1.Often processing is inoculated with 30 explants or so, is repeated 3 times, often
Bottle 3 explants of inoculation.Callus is obtained after cultivating 30 days.Wherein, condition of culture is：Intensity of illumination 1700Lx, 14h light
According to/10h dark, temperature is to carry out dark culturing in 25 ± 2 DEG C of tissue culture room；Routine observation, count callus induction rate.
Analysis of experimental results：As shown in table 5, under same breeding condition, callus lures the explant of different cultivars
Conductance difference is obvious, and the inductivity highest of the present embodiment " Haas " avocado stem section explant, and the explant of other kinds
Inductivity is far below " Haas " avocado, and this absolutely proves that different genotype explant has difference to the sensitiveness of plant growth regulator
It is different, it is different from different explant type evoked callus abilities, may with it hormone-content it is relevant.
Influence of the 6 same inducing culture of table to the callus induction of the explant of different cultivars
Above-described is only the preferred embodiment of the present invention, it is noted that for one of ordinary skill in the art
For, without departing from the concept of the premise of the invention, various modifications and improvements can be made, these belong to the present invention
Protection domain.
Claims (10)
1. a kind of abductive approach of avocado callus, it is characterised in that comprise the following steps：
Avocado explant is inoculated into inducing culture, in 1200~1700Lx of intensity of illumination, 8~14h/ days photoperiod, temperature
Cultivated 4~30 days under conditions of 22~28 DEG C of degree, obtain callus, wherein, the inducing culture is to contain 20~40g/L
Sucrose, 5~12g/L agar, 0~3.0mg/L methyl α-naphthyl acetates, 0.1~3.0mg/L 2,4 dichlorophenoxyacetic acids, 0.1~3.0mg/L
The MS basal mediums that 6- benzyls aminoadenine and pH are 5.8~6.5.
2. the abductive approach of avocado callus as claimed in claim 1, it is characterised in that the avocado explant includes oil
One or more in Pear leaves, avocado petiole, avocado stem apex, avocado stem section, avocado bud section.
3. the abductive approach of avocado callus as claimed in claim 1, it is characterised in that the kind of the avocado includes
" Haas " avocado, " Reed " avocado, " one kind in the avocados of Duke 7 ", " A10 " avocado.
4. the abductive approach of avocado callus as claimed in claim 1, it is characterised in that the training of the avocado callus
The time is educated for 6~14 days.
5. the abductive approach of avocado callus as claimed in claim 1, it is characterised in that the inducing culture be containing
20~40g/L sucrose, 5~12g/L agar, 0.1~3.0mg/L 2,4 dichlorophenoxyacetic acids, 0.1~3.0mg/L6- benzyl amino
The MS basal mediums that adenine and pH are 5.8~6.5.
6. the abductive approach of avocado callus as claimed in claim 1, it is characterised in that the inducing culture be containing
20~40g/L sucrose, 5~12g/L agar, 0.1~2.0mg/L 6- benzyls aminoadenine, 0.1mg/L 2,4 dichloro benzene oxygen second
The MS basal mediums that acid and pH are 5.8~6.5.
7. the abductive approach of avocado callus as claimed in claim 1, it is characterised in that the avocado callus lures
In guiding method, the inductivity of the avocado callus is not less than 75%.
8. the abductive approach of avocado callus as claimed in claim 1, it is characterised in that the explant is to collect
Avocado sample sterilization be made.
A kind of 9. method for promoting the differentiation of avocado callus bud, it is characterised in that comprise the following steps：
The callus for taking the abductive approach of the avocado callus as described in claim 1~8 is any to cultivate, by being cured for gained
Injured tissue is inoculated into bud differential medium and cultivated 30~50 days, and the bud differential medium is to contain 20~40g/L sucrose, 5
~12g/L agar, 0.1~3.0mg/L methyl α-naphthyl acetates, 0.1~3.0mg/L 6- benzyls aminoadenines and pH are 5.8~6.5 MS
Basal medium.
10. promote the method for avocado callus bud differentiation as claimed in claim 9, it is characterised in that the avocado callus
The bud ratio of tissue is not less than 35~60%.
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| CN107410024B CN107410024B (en) | 2020-06-02 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108308030A (en) * | 2018-04-04 | 2018-07-24 | 中国热带农业科学院南亚热带作物研究所 | A method of promoting avocado stem section evoked callus and sterile bud |
| CN108496802A (en) * | 2018-04-17 | 2018-09-07 | 沈阳农业大学 | A kind of pear cv nanguo callus, cultural method and application |
| CN111543319A (en) * | 2020-04-21 | 2020-08-18 | 中国热带农业科学院海口实验站 | In-vitro rescue method for immature embryo of avocado |
| CN114711145A (en) * | 2022-05-10 | 2022-07-08 | 北京林业大学 | Tissue culture method of potentilla glandulifera |
-
2017
- 2017-05-16 CN CN201710345217.XA patent/CN107410024B/en not_active Expired - Fee Related
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108308030A (en) * | 2018-04-04 | 2018-07-24 | 中国热带农业科学院南亚热带作物研究所 | A method of promoting avocado stem section evoked callus and sterile bud |
| CN108496802A (en) * | 2018-04-17 | 2018-09-07 | 沈阳农业大学 | A kind of pear cv nanguo callus, cultural method and application |
| CN111543319A (en) * | 2020-04-21 | 2020-08-18 | 中国热带农业科学院海口实验站 | In-vitro rescue method for immature embryo of avocado |
| CN111543319B (en) * | 2020-04-21 | 2021-07-30 | 中国热带农业科学院海口实验站 | In-vitro rescue method for immature embryo of avocado |
| CN114711145A (en) * | 2022-05-10 | 2022-07-08 | 北京林业大学 | Tissue culture method of potentilla glandulifera |
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| CN107410024B (en) | 2020-06-02 |
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