CN109548652A - Tillandsia callus and its cultural method and application - Google Patents
Tillandsia callus and its cultural method and application Download PDFInfo
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- CN109548652A CN109548652A CN201811483960.2A CN201811483960A CN109548652A CN 109548652 A CN109548652 A CN 109548652A CN 201811483960 A CN201811483960 A CN 201811483960A CN 109548652 A CN109548652 A CN 109548652A
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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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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- Developmental Biology & Embryology (AREA)
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Abstract
The invention discloses a kind of cultural method of Tillandsia callus and Tillandsia quick-breeding methods.The cultural method of the Tillandsia callus includes the following steps have: obtaining the organization of root tips of Tillandsia, and carries out disinfection processing to the organization of root tips；The organization of root tips after will be sterile-processed is seeded to progress Fiber differentiation processing in induced medium, obtains callus；The callus is transferred to progress shoot proliferation culture processing in subculture multiplication medium.The explant material of Tillandsia callus culture method of the present invention easily obtains, zero injury basic to former plant；By the Fiber differentiation processing and shoot proliferation culture processing to organization of root tips, a large amount of callus can be obtained, and there is stable differentiation capability.Tillandsia quick-breeding method of the present invention can be realized the quick and a large amount of breeding of Tillandsia, and the Tillandsia character bred is stablized.
Description
Technical field
The invention belongs to field of plant tissue culture technique, and in particular to a kind of Tillandsia callus and its culture side
Method and application.
Background technique
Tillandsia is under the jurisdiction of pineapple family (Bromeliaceae), is monocotyledon, perennial herb.Pineapple family has 3
A subfamily, 50 categories including nearly 550 kinds and 90 mutation.3 subfamilies are respectively Honoka pineapple subfamily
(Pitcairnioideae), iron Lan Yake (Tillandsioideae) and pineapple subfamily (Bromelioideae).Wherein, fox
Leopard cat tail Tillandsia (Tillandsia Funkiana) is common one kind in Tillandsia, is that typical gas is raw or grow nonparasitically upon another plant
Plant.
The main nutrient and the organ of moisture of absorbing of Tillandsia is leaf surface covering " sunflower shape " scale structure.The knot
Structure can help Tillandsia directly to obtain nutrient and moisture from the air there are pollutant.And their root system is degenerated, and is only existed
Fixed function is played on supporter, does not need soil, so they do not have any contact with ground.
Tillandsia is can to admire the beauty of flowers and the ornamental plant of leaf.Tillandsia is various in style, and plant shape is graceful, decoration
Property is extremely strong.It can with individual character be pasted on ancient tree stake, rockwork and wall, or be placed in bamboo basket, shell, intersperse space, fashion
It is pure and fresh, rich in the Nature rustic charm；Composition potted landscape can also be spelled, be decorated screen, room can be allowed vigorous.In addition to this,
Tillandsia also has the advantages that viewing period long, and almost whole year can bloom；Tillandsia sets flower without cultivation matrix, without taking
Frame, sanitation and hygiene can arbitrarily be put, and be ideal decorative indoor plant.Tillandsia simultaneously can be used for ornamental plant
Scape, green landscape group scape are made, or holds Tillandsia plant special topic flower show in festivals or holidays.Introduced a fine variety in recent years to the country, rapidly at
New lover and novel indoor appreciation flowers for plant fan, step into common people house.
At present Tillandsia breeding mostly use traditional division propagation and cutting propagation greatly, it is also possible to tissue cultures and at
Seed in the ripe fruit pod that do not crack carries out sterile culture.Seminal propagation success rate is high, but when needing 5-8 by seed growth to finished product
Between, and seed production is few；Though plant fast propagation speed is fast, since Tillandsia blade is covered with scale to the disinfection band of explant
Carry out very big difficulty；It is sent out fastly using terminal bud or lateral bud as explant, can frequently result in mould contamination and failure；This external cause air phoenix
Pears differentiation degree is high, and evoked callus is also more difficult, therefore still belongs to the exploratory stage to the tissue cultures of Tillandsia at present
Therefore flowers manufacturer mostly uses division propagation at present.How effectively to breed Tillandsia is that those skilled in the art make great efforts to solve always
Technical problem certainly.
Summary of the invention
It is an object of the invention to overcome the above-mentioned deficiency of the prior art, a kind of Tillandsia callus and its training are provided
It is undesirable to obtain difficult and reproductive efficiency with the explant material that solution is currently used in Tillandsia breeding for the method for supporting and application
The technical issues of.
In order to achieve the above-mentioned object of the invention, an aspect of of the present present invention provides a kind of culture of Tillandsia callus
Method.The cultural method of the Tillandsia callus includes the following steps:
The organization of root tips of Tillandsia is obtained, and is carried out disinfection processing to the organization of root tips；
The organization of root tips after will be sterile-processed is seeded to progress Fiber differentiation processing in induced medium, is cured
Injured tissue；
The callus is transferred to progress shoot proliferation culture processing in subculture multiplication medium.
Another aspect of the present invention provides a kind of Tillandsia callus.The Tillandsia callus is by this
The cultural method culture of invention Tillandsia callus obtains.
Another aspect of the invention provides a kind of Tillandsia quick-breeding method.The Tillandsia quick-breeding method includes
Tillandsia callus of the present invention is subjected to the step of differentiation culture becomes seedling.
Compared with prior art, the cultural method of Tillandsia callus of the present invention uses the organization of root tips of Tillandsia
It as explant material, easily obtains, zero injury basic to former plant is disinfected simple and easy；On the other hand, by root
The Fiber differentiation processing and shoot proliferation culture processing of point tissue, can obtain a large amount of callus, and have stable differentiation
Ability.
Tillandsia callus of the present invention is obtained by the cultural method culture of Tillandsia callus of the present invention, because
This, the Tillandsia callus can mass propgation, and it is with stable differentiation capability.
Tillandsia quick-breeding method of the present invention, which directly carries out differentiation culture using Tillandsia callus of the present invention, to be become
Seedling, therefore, the Tillandsia quick-breeding method can be realized the quick and a large amount of breeding of Tillandsia, and the sky bred
Gas pineapple character is stablized.
Detailed description of the invention
Fig. 1 is the cultural method process flow chart of Tillandsia callus provided in an embodiment of the present invention.
Specific embodiment
In order to which technical problems, technical solutions and advantageous effects to be solved by the present invention are more clearly understood, below in conjunction with
Embodiment, the present invention will be described in further detail.It should be appreciated that specific embodiment described herein is only used to explain
The present invention is not intended to limit the present invention.
Unless otherwise defined, otherwise all technical terms and scientific terms used herein have and institute of the embodiment of the present invention
Belong to technical field those of ordinary skill and is generally understood identical meaning.If the definition stated in this part be incorporated by reference
The definition stated in the patent of this paper, patent application, the patent application of announcement and other publications is opposite or other aspects
Inconsistent, the definition listed in this part is prior to the definition in being totally incorporated herein by reference.
On the one hand, the embodiment of the present invention provides a kind of cultural method of Tillandsia callus.The Tillandsia is cured
The cultural method process flow of injured tissue is as shown in Figure 1, the cultural method includes the following steps:
S01. the disinfection of explant: obtaining the organization of root tips of Tillandsia, and carries out disinfection processing to the organization of root tips；
S02. calli induction: the organization of root tips after will be sterile-processed, which is seeded in induced medium, to be carried out
Fiber differentiation processing, obtains callus；
S03. the shoot proliferation culture of callus: by the callus be transferred in subculture multiplication medium carry out after
For Multiplying culture processing.
Wherein, it in above-mentioned steps S01, using the organization of root tips of Tillandsia as explant material, easily obtains, it is right
Former basic zero injury of plant, and disinfect simple and easy.In one embodiment, the organization of root tips is Tillandsia root system
The 0.5-2cm tip of a root.The preferably 0.5-2cm tip of a root of the vigorous root system of growth of Tillandsia.
In another embodiment, the disinfection treatment of the organization of root tips is included the following steps:
It is first cleaned with aqua sterilisa and 10-15min is impregnated to the organization of root tips, then be transferred to 0.1% HgCl2It is sterilized in solution
3-5min is handled, then aqua sterilisa rinses 3-5 times.
In addition, the Tillandsia can be common haretail uraria herb Tillandsia.
In above-mentioned steps S02, the organization of root tips, which is seeded in induced medium, carries out Fiber differentiation processing, and described
The cell of sharp tissue tissue part can occur cell division and be proliferated to form callus.In one embodiment, the Fiber differentiation
Base includes following component:
MS minimal medium, 2,4 dichlorophenoxyacetic acid 0.2-1.5mg/L, diethylamino ethanol caproate 0-2mg/L.
By adding suitable 2,4 dichlorophenoxyacetic acid and hexanoic acid diethylaminoethanol in the MS minimal medium
Ester can effectively induce the cell division of the organization of root tips to be proliferated to form callus.
Wherein, the organization of root tips in step S01 after disinfection treatment is placed in the induced medium and is induced
Culture can grow callus in organization of root tips.In one embodiment, the organization of root tips after disinfection treatment is placed in institute
Stating the progress Fiber differentiation time in induced medium is preferably 2-6 weeks.Condition of culture is preferred are as follows: and 25-30 DEG C of temperature, illumination 12~
16h/ days, 1500~2000lx of illuminance.In above-mentioned steps S03, callus described in step S03 is transferred to subculture and is increased
Grow the continuous multi-generation culture that culture medium realizes callus.In one embodiment, the subculture multiplication medium includes such as the following group
Point:
MS minimal medium, 6- benzyl aminoadenine 0.1-1.5mg/L, 2,4 dichlorophenoxyacetic acid 0.8-1.5mg/L.
By adding suitable 6- benzyl aminoadenine and 2,4 dichlorophenoxyacetic acid energy in the MS minimal medium
Big callus is measured in enough continuous multi-generation Multiplying cultures for effectively promoting to induce the callus, acquisition, and improves described be cured
The stable differentiation capability of injured tissue.
In a further embodiment, in the step of shoot proliferation culture is handled, every 30-40 days to the callus
Tissue transfer primary.Switching subculture multiplication medium used contains 6- benzyl aminoadenine and 2,4 dichloro benzene to be above-mentioned
The shoot proliferation culture of fluoroacetic acid.
Wherein, the step S02 callus cultivated is placed in the shoot proliferation culture and carries out shoot proliferation culture
Time can be adjusted and control according to the demand of callus, in one embodiment, shoot proliferation condition of culture are as follows: temperature
25-30 DEG C of degree, illumination 12~16h/ days, 1500~2000lx of illuminance.To improve the shoot proliferation of callus.
Therefore, the cultural method of the Tillandsia callus is using the organization of root tips of Tillandsia as explant material
Material, so that explant material is relatively easily obtained, does not injure former plant substantially, easily disinfects；On the other hand, by excellent
Change induced medium and shoot proliferation culture, realizes and the Fiber differentiation processing and shoot proliferation culture of organization of root tips are handled, from
And a large amount of callus can be obtained, and assign the callus stable differentiation capability.In addition, the Tillandsia callus
The cultural method condition of tissue is easily-controllable, and the Tillandsia callus character of acquisition is stablized, and differentiation capability is strong, and high-efficient.
On the other hand, on the basis of the cultural method of Tillandsia callus described above, the embodiment of the present invention is also
Provide a kind of Tillandsia callus.The training of Tillandsia callus Tillandsia callus by mentioned earlier
Method culture is supported to obtain.Therefore, the Tillandsia callus can mass propgation, and it is with stable differentiation potency
Power.
In another aspect, on the basis of Tillandsia callus described above and its cultural method, the embodiment of the present invention
Additionally provide a kind of Tillandsia quick-breeding method.The Tillandsia quick-breeding method includes by Tillandsia callus group described above
It knits and carries out the step of differentiation culture becomes seedling.Therefore, the Tillandsia quick-breeding method can be realized the quick of Tillandsia
And a large amount of breeding, and the Tillandsia character bred is stablized, and reduces costs.
Now by taking specific Tillandsia callus and its cultural method as an example, the present invention will be described in further detail.
Embodiment 1
Present embodiments provide a kind of Tillandsia callus and its cultural method.The Tillandsia callus training
Feeding method includes the following steps:
S11. the disinfection of explant:
Haretail uraria herb Tillandsia (Tillandsia Funkiana) eugonic root system is chosen, cutting length is 1.0cm
The tip of a root is cleaned with aqua sterilisa, is impregnated 15min, is transferred to 0.1% HgCl23min is disinfected in solution, aqua sterilisa rinses 5 times；
S12. calli induction:
The tip of a root after step S11 disinfection treatment is seeded on MS minimal medium, one tip of a root of every bottle of inoculation, and added
2,4- dichlorphenoxyacetic acid 1.0mg/L and diethylamino ethanol caproate 0.5mg/L carry out culture processing, 18-22 days root tips
Position cell division is proliferated to form callus, callus rate 80%；
S13. shoot proliferation culture:
The callus formed in step S12 is transferred to be transferred to and is added to 6- benzyl aminoadenine 0.5mg/L and 2,4-
Shoot proliferation culture is carried out in the MS minimal medium of dichlorphenoxyacetic acid 0.8mg/L, switching in every 30-40 days is primary, obtains big
Measure callus.
Embodiment 2
Present embodiments provide a kind of Tillandsia callus and its cultural method.The Tillandsia callus training
Feeding method includes the following steps:
S11. the disinfection of explant:
Haretail uraria herb Tillandsia (Tillandsia Funkiana) eugonic root system is chosen, cutting length is 0.5cm
The tip of a root is cleaned with aqua sterilisa, is impregnated 15min, is transferred to 0.1% HgCl24min is disinfected in solution, aqua sterilisa rinses 5 times；
S12. calli induction:
The tip of a root after step S11 disinfection treatment is seeded on MS minimal medium, one tip of a root of every bottle of inoculation, and added
2,4- dichlorphenoxyacetic acid 1.2mg/L and diethylamino ethanol caproate 0.5mg/L carry out culture processing, 15-20 days root tips
Position cell division is proliferated to form callus, callus rate 73%；
S13. shoot proliferation culture:
The callus formed in step S12 is transferred to be transferred to and is added to 6- benzyl aminoadenine 0.8mg/L and 2,4-
Shoot proliferation culture is carried out in the MS minimal medium of dichlorphenoxyacetic acid 1.2mg/L, switching in every 30-40 days is primary, obtains big
Measure callus.
Embodiment 3
Present embodiments provide a kind of Tillandsia callus and its cultural method.The Tillandsia callus training
Feeding method includes the following steps:
S11. the disinfection of explant:
Haretail uraria herb Tillandsia (TillandsiaFunkiana) eugonic root system is chosen, cutting length is 1.0cm
The tip of a root is cleaned with aqua sterilisa, is impregnated 10min, is transferred to 0.1% HgCl25min is disinfected in solution, aqua sterilisa rinses 5 times；
S12. calli induction:
The tip of a root after step S11 disinfection treatment is seeded on MS minimal medium, one tip of a root of every bottle of inoculation, and added
2,4- dichlorphenoxyacetic acid 1.2mg/L and diethylamino ethanol caproate 1.0mg/L carry out culture processing, 15-18 days root tips
Position cell division is proliferated to form callus, callus rate 92%；
S13. shoot proliferation culture:
The callus formed in step S12 is transferred to be transferred to and is added to 6- benzyl aminoadenine 0.4mg/L and 2,4-
Shoot proliferation culture is carried out in the MS minimal medium of dichlorphenoxyacetic acid 0.8mg/L, switching in every 30-40 days is primary, obtains big
Measure callus.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Made any modifications, equivalent replacements, and improvements etc., should all be included in the protection scope of the present invention within mind and principle.
Claims (9)
1. a kind of cultural method of Tillandsia callus, which is characterized in that the cultural method includes the following steps:
The organization of root tips of Tillandsia is obtained, and is carried out disinfection processing to the organization of root tips；
The organization of root tips after will be sterile-processed is seeded to progress Fiber differentiation processing in induced medium, obtains callus group
It knits；
The callus is transferred to progress shoot proliferation culture processing in subculture multiplication medium.
2. cultural method according to claim 1, which is characterized in that the induced medium includes following component:
MS minimal medium；
2,4 dichlorophenoxyacetic acid 0.2-1.5mg/L；
Diethylamino ethanol caproate 0-2mg/L.
3. cultural method according to claim 1, which is characterized in that the subculture multiplication medium includes following component:
MS minimal medium；
6- benzyl aminoadenine 0.1-1.5mg/L；
2,4 dichlorophenoxyacetic acid 0.8-1.5mg/L.
4. cultural method according to claim 3, which is characterized in that in the step of shoot proliferation culture is handled,
The callus transfer every 30-40 days primary.
5. cultural method according to claim 1-4, which is characterized in that the organization of root tips is Tillandsia root
The 0.5-2cm tip of a root of system.
6. cultural method according to claim 5, which is characterized in that the Tillandsia is haretail uraria herb Tillandsia.
7. -4,6 described in any item cultural methods according to claim 1, which is characterized in that carry out disinfection to the organization of root tips
Processing includes the following steps:
It is first cleaned with aqua sterilisa and 10-15min is impregnated to the organization of root tips, then be transferred to 0.1% HgCl2It is disinfected in solution
3-5min, then aqua sterilisa rinses 3-5 times.
8. a kind of Tillandsia callus, which is characterized in that the Tillandsia callus is by any institute of claim 1-7
The cultural method culture stated obtains.
9. a kind of Tillandsia quick-breeding method, which is characterized in that including by Tillandsia callus according to any one of claims 8 into
The step of row differentiation culture becomes seedling.
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