CN109006488A - A kind of red luxuriant pineapple breeding method based on using buds to propagate buds - Google Patents
A kind of red luxuriant pineapple breeding method based on using buds to propagate buds Download PDFInfo
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- CN109006488A CN109006488A CN201811213854.2A CN201811213854A CN109006488A CN 109006488 A CN109006488 A CN 109006488A CN 201811213854 A CN201811213854 A CN 201811213854A CN 109006488 A CN109006488 A CN 109006488A
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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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Abstract
The red luxuriant pineapple breeding method that the invention discloses a kind of based on using buds to propagate buds, use tissue cultures entirely green plant for material, after being acquired on callus, it is transplanted to Rooting and hardening-off culture in culture medium, after stem top cuts out top processing, strong sprout is moved into dark surrounds by the elongation culture of 40d stem again, stem length >=3cm plant is transferred to culture induction lateral bud under illumination condition to sprout, re-segmenting, which moves into, is conducive to squamous subculture in the culture medium that bud is sprouted and takes root, and finally obtains and the consistent regeneration plant of maternal plant character.The present invention obtains complete regenerated plant by way of in vitro culture, overcomes in previous red luxuriant pineapple tissue culture procedures, by callus again the organ genetic method of atomization, the high disadvantage of aberration rate ensure that the consistency between regeneration plant and maternal plant.
Description
Technical field
The invention belongs to red luxuriant pineapple Cultivating techniques fields, specifically, being related to a kind of red luxuriant phoenix based on using buds to propagate buds
Pear cultivating method.
Background technique
The prior art during establishing Propagation In Vitro system, can pass through callus using red luxuriant pineapple as material
Differentiation pathway again, the variation character of adventitious bud are up to 99%, and variation character is more and irregular, the predominantly complete green plant of variation type
With complete white plant, in addition there are also chimeric plants；The about complete green plant 43% of these three plant proportions, entirely white plant 38%,
Chimeric plant 19%, wherein chimeric character is widely different (Fig. 1).Therefore, to obtain red luxuriant pineapple Chimera Variety, pass through callus
The approach of regeneration adventitious bud is bred, and is worthless.
It is therefore desirable to provide a kind of new red luxuriant pineapple breeding method.
Summary of the invention
In view of this, the red luxuriant pineapple breeding method that the present invention provides a kind of based on using buds to propagate buds.
In order to solve the above-mentioned technical problem, the red luxuriant pineapple breeding method that the invention discloses a kind of based on using buds to propagate buds,
Using tissue cultures, green plant after acquiring on callus, is transplanted to Rooting and hardening-off culture in culture medium, in stem for material entirely
After top processing is cut out on top, then strong sprout is moved into dark surrounds and extends culture by 40d stem, stem length >=3cm plant is transferred to
Culture induction lateral bud is sprouted under illumination condition, and re-segmenting, which moves into, is conducive to squamous subculture in the culture medium that bud is sprouted and takes root, finally
It obtains and the consistent regeneration plant of maternal plant character.
Optionally, comprising the following steps:
Step 1, material select: acquisition passes through the regeneration that tissue cultures obtain and plants using red luxuriant pineapple stem section as explant
Strain is material；Plantlet is cut from callus, is vertically transferred on MS culture medium, strong seedling culture；It is long to 3cm- to seedling
4cm, stem length to 0.5cm, and several 2-4 items of taking root, it is spare；
Step 2 cuts out top processing: the plant after strong seedling culture, in stem top cutting, eliminates plant under super-clean bench
Apical dominance is transferred in MS culture medium and cultivates；
Step 3, dark culture induce internode elongation: the plant after strong sprout cuts out top is put into carton raising middle flask and carries out dark culture；
Plant internode elongation length 1cm-4cm after dark culture；
Step 4, illumination cultivation: after terminating the dark phase, stem elongation >=3cm plant is removed into carton, is made for a long time in black
Plant in dark situation carries out bud under illumination condition and sprouts induction；
Step 5, segmentation squamous subculture: by the plant after bud sprouts induction, it is cut into the segment saved with 2-3, respectively
For terminal bud section, middle section, band foundation section, vertically it is seeded on culture medium induces axillary bud sprouting respectively；
The formation of step 6, regeneration plant: after segmentation squamous subculture 1.5 months, axillary bud can be sprouted on section, meeting after 2 months
Start to take root in base portion, forms intact plant after 3-4 months；
Step 7, plantlet of transplant: intact plant is gently cut from maternal plant, is transplanted on MS culture medium or is directly transplanted
Into matrix, plantlet of transplant is completed.
Optionally, the plantlet in the step 1 is growing way health, the plant that the number of blade is 5-8 piece.
Optionally, the strong seedling culture condition in the step 1 is the strong seedling culture under conditions of 25 ± 2 DEG C, illumination 12h/d
28 days.
Optionally, in the step 2 in MS culture medium condition of culture are as follows: under conditions of 25 ± 2 DEG C, illumination 12h/d
Culture one week.
Optionally, the dark culture condition in the step 3 are as follows: 25 ± 2 DEG C of cultivation temperature；Incubation time 40d, during which every
10d takes out illumination 1h.
Optionally, it is illumination cultivation that bud in the step 4, which sprouts inductive condition, specifically: at 25 ± 2 DEG C, illumination
15d is cultivated under conditions of 12h/d.
Optionally, the culture medium of the segmentation squamous subculture in the step 5 is MS+BA3mg/l+NAA2mg/l.
Optionally, the matrix in the step 7 includes the river sand and coco bran that mass ratio is 1:1.
Compared with prior art, the present invention can be obtained including following technical effect:
1) present invention is auxiliary using method for tissue culture using the complete green plant differentiated again by callus as material
With suitable temperature, red luxuriant pineapple internode elongation is induced by dark culture, is segmented subculture on Optimal Medium, induction lateral bud is sprouted
Hair, takes root for forming intact plant, by normally cultivating the case where 0 sprouting, germination rate is made to be up to 62% on section.
2) present invention obtains complete regenerated plant by way of in vitro culture, overcomes previous red luxuriant pineapple tissue cultures
In the process, by callus again the organ genetic method of atomization, the high disadvantage of aberration rate ensure that regeneration plant and mother
Consistency between strain.
Certainly, it implements any of the products of the present invention it is not absolutely required to while reaching all the above technical effect.
Detailed description of the invention
The drawings described herein are used to provide a further understanding of the present invention, constitutes a part of the invention, this hair
Bright illustrative embodiments and their description are used to explain the present invention, and are not constituted improper limitations of the present invention.In the accompanying drawings:
Fig. 1 is the adventitious bud of different chimeric characters in background of invention；Wherein, A represents callus primary differentiation
Full green and whole white make a variation tissue-cultured seedling, B represents irregular chimera, and C represents the chimera (Phnom Penh) of leaf margin white, D generation
Table leaf margin green, middle part have the chimera (the golden heart) of white stripes, and E represents irregular chimera blade, and F represents irregular
Chimera blade；
Fig. 2 is the aspect graph for strengthening green seedling in the selection of material of the present invention；
Fig. 3 is the aspect graph of the complete green seedling of stem elongation after dark culture of the present invention；
Fig. 4 is strong sprout and excessive growth seedling comparison diagram after dark culture before dark culture of the present invention, wherein A represents strong sprout before dark culture,
B represents excessive growth seedling after dark culture；
Fig. 5 is the aspect graph of axil director normal bud, wherein axil director is big after A is segmented subculture after present invention segmentation subculture
The normal bud to take root, children tender normal bud when B is segmented subculture；
Fig. 6 is the aspect graph that the present invention saves upper axillary bud.
Specific embodiment
Carry out the embodiment that the present invention will be described in detail below in conjunction with embodiment, whereby to the present invention how application technology hand
Section solves technical problem and reaches the realization process of technical effect to fully understand and implement.
Embodiment 1
A kind of red luxuriant pineapple breeding method based on using buds to propagate buds, comprising the following steps: the culture in strong sprout stage is cut out at top
Reason, the processing of dark culture, the processing of illumination cultivation, segmentation subculture induce the culture of initial bud, strengthen bud culture and culture of rootage work
Sequence；Using tissue cultures, green plant after acquiring on callus, is transplanted to Rooting and hardening-off culture in culture medium for material entirely,
It moves into after stem top cuts out top processing, then by strong sprout by the elongation culture of 40d stem in dark surrounds, by stem length >=3cm plant
Culture induction lateral bud under illumination condition is transferred to sprout, re-segmenting, which moves into, is conducive to squamous subculture in the culture medium that bud is sprouted and takes root,
Finally acquisition and the consistent regeneration plant of maternal plant character, specific steps are as follows:
Step 1, material select: acquisition passes through the regeneration that tissue cultures obtain and plants using red luxuriant pineapple stem section as explant
Strain is material；The plantlet that growing way health, the number of blade about 5-8 piece are cut from callus, is vertically transferred on MS culture medium,
Strong seedling culture 28 days under conditions of 25 ± 2 DEG C, illumination 12h/d；It is about to seedling to 3cm-4cm, stem length is and raw about to 0.5cm
Radical 2-4 item, as shown in Fig. 2, spare；
Step 2 cuts out top processing: the plant after strong seedling culture, in stem top cutting, eliminates plant under super-clean bench
Apical dominance is transferred to MS culture medium, cultivates one week under conditions of 25 ± 2 DEG C, illumination 12h/d；
Step 3, dark culture induce internode elongation: the plant after strong sprout cuts out top is put into carton raising middle flask and carries out dark culture,
25 ± 2 DEG C of cultivation temperature；During which incubation time 40d takes out illumination 1h every 10d.Plant internode after dark culture is bright
Aobvious elongation, extended length about 1cm-4cm (Fig. 3), state at this time are conducive to the formation and development of axillary bud；Strong sprout before dark culture
Fig. 4 is seen with excessive growth seedling comparison after dark culture, and by Fig. 4 as it can be seen that after dark culture, plant internode has obvious elongation.
1 dark culture 40d stem length degree situation of table compares
| Stem extended length | 1cm | 2cm | 3cm | 4cm |
| Stem stretches ratio | 4% | 12% | 36% | 48% |
Note: stem elongation plant sum after plant number/dark culture of different stem extended lengths after the stem ratio of stretching=dark culture.
Step 4, illumination cultivation: after terminating the dark phase, stem elongation >=3cm plant is removed into carton, is made for a long time in black
Plant in dark situation is in normal health growth conditions, and 15d is cultivated under conditions of 25 ± 2 DEG C, illumination 12h/d, carries out bud
Sprout induction；
Step 5, segmentation squamous subculture: by the plant after illumination cultivation, a plant be cut into band 2-3 section it is small
Section, respectively terminal bud section, middle section, band foundation section, are vertically seeded in respectively on the culture medium of MS+BA3mg/l+NAA 2mg/l
Axillary bud sprouting is induced, wherein band foundation section axillary bud sprouting rate highest, up to 68%；Segmentation subculture axillary bud sprouting rate is shown in Table 2.
Table 2 is segmented subculture axillary bud sprouting rate
The formation of step 6, regeneration plant: after segmentation squamous subculture about 1.5 months, axillary bud can be sprouted on section, after 2 months
It can start to take root in base portion, form intact plant after 3-4 months；
3 regeneration plant of table sprouts number and germination rate
Step 7, plantlet of transplant: intact plant is gently cut from maternal plant, is transplanted on MS culture medium or is directly transplanted
Into the matrix containing river sand and coco bran that mass ratio is 1:1, i.e., viable, survival rate > 95%.
Above description has shown and described several preferred embodiments of invention, but as previously described, it should be understood that invention is not
It is confined to form disclosed herein, should not be regarded as an exclusion of other examples, and can be used for various other combinations, modification
And environment, and can be carried out within that scope of the inventive concept describe herein by the above teachings or related fields of technology or knowledge
Change.And changes and modifications made by those skilled in the art do not depart from the spirit and scope of invention, then it all should be in the appended power of invention
In the protection scope that benefit requires.
Claims (9)
1. a kind of red luxuriant pineapple breeding method based on using buds to propagate buds, which is characterized in that use tissue cultures entirely green plant for material
Material is transplanted to Rooting and hardening-off culture in culture medium after acquiring on callus, after stem top cuts out top processing, then by strong sprout
It moves into dark surrounds by the elongation culture of 40d stem, stem length >=3cm plant is transferred to culture induction lateral bud under illumination condition and is sprouted
Hair, re-segmenting, which moves into, is conducive to squamous subculture in the culture medium that bud is sprouted and takes root, and finally obtains and the consistent regeneration of maternal plant character
Plant.
2. red luxuriant pineapple breeding method according to claim 1, which comprises the following steps:
Step 1, material select: acquisition is by the regeneration plant that tissue cultures obtain using red luxuriant pineapple stem section as explant
Material；Plantlet is cut from callus, is vertically transferred on MS culture medium, strong seedling culture；It is long to 3cm-4cm, stem to seedling
It grows to 0.5cm, and several 2-4 items of taking root, it is spare；
Step 2 cuts out top processing: the plant after strong seedling culture, in stem top cutting, eliminates the top of plant under super-clean bench
Advantage is transferred in MS culture medium and cultivates；
Step 3, dark culture induce internode elongation: the plant after strong sprout cuts out top is put into carton raising middle flask and carries out dark culture；By
Plant internode elongation length 1cm-4cm after dark culture；
Step 4, illumination cultivation: after terminating the dark phase, removing carton for stem elongation >=3cm plant, makes for a long time in dark ring
Plant in border carries out bud under illumination condition and sprouts induction；
Step 5, segmentation squamous subculture: by the plant after bud sprouts induction, it is cut into the segment saved with 2-3, is respectively pushed up
Bud section, middle section, band foundation section, are vertically seeded on culture medium induce axillary bud sprouting respectively；
The formation of step 6, regeneration plant: after segmentation squamous subculture 1.5 months, axillary bud can be sprouted on section, can start after 2 months
It takes root in base portion, forms intact plant after 3-4 months；
Step 7, plantlet of transplant: intact plant is gently cut from maternal plant, is transplanted on MS culture medium or is directly transplanted to base
In matter, plantlet of transplant is completed.
3. red luxuriant pineapple breeding method according to claim 2, which is characterized in that the plantlet in the step 1 is length
Gesture health, the plant that the number of blade is 5-8 piece.
4. red luxuriant pineapple breeding method according to claim 2, which is characterized in that the strong seedling culture item in the step 1
Part is strong seedling culture 28 days under conditions of 25 ± 2 DEG C, illumination 12h/d.
5. red luxuriant pineapple breeding method according to claim 2, which is characterized in that in the step 2 in MS culture medium
Condition of culture are as follows: cultivated one week under conditions of 25 ± 2 DEG C, illumination 12h/d.
6. red luxuriant pineapple breeding method according to claim 2, which is characterized in that the dark culture condition in the step 3
Are as follows: 25 ± 2 DEG C of cultivation temperature；During which incubation time 40d takes out illumination 1h every 10d.
7. red luxuriant pineapple breeding method according to claim 2, which is characterized in that the bud in the step 4 sprouts induction
Condition is illumination cultivation, specifically: 15d is cultivated under conditions of 25 ± 2 DEG C, illumination 12h/d.
8. red luxuriant pineapple breeding method according to claim 2, which is characterized in that segmentation in the step 5 is after being commissioned to train
Feeding culture medium is MS+BA3mg/l+NAA 2mg/l.
9. red luxuriant pineapple breeding method according to claim 2, which is characterized in that the matrix in the step 7 includes matter
Amount is than the river sand and coco bran for 1:1.
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| CN201811213854.2A CN109006488A (en) | 2018-10-15 | 2018-10-15 | A kind of red luxuriant pineapple breeding method based on using buds to propagate buds |
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| CN201811213854.2A CN109006488A (en) | 2018-10-15 | 2018-10-15 | A kind of red luxuriant pineapple breeding method based on using buds to propagate buds |
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Citations (5)
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|---|---|---|---|---|
| WO2004052085A1 (en) * | 2002-12-06 | 2004-06-24 | Del Monte Fresh Produce Company | Transgenic pineapple plants with modified carotenoid levels and methods of their production |
| CN101803570A (en) * | 2010-03-26 | 2010-08-18 | 浙江传化生物技术有限公司 | Method for efficiently propagating torch pineapples by utilizing in-vitro leaves |
| WO2013043508A1 (en) * | 2011-09-20 | 2013-03-28 | The Texas A&M University System | Citrus shoot regeneration compositions, methods, and systems |
| CN105028194A (en) * | 2015-06-23 | 2015-11-11 | 青岛农业大学 | Tissue culture rapid propagation method of tillandsia |
| CN106613940A (en) * | 2016-10-05 | 2017-05-10 | 南宁邃丛赋语科技开发有限责任公司 | In-vitro rapid propagation method for billbergia pyramidalis |
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2018
- 2018-10-15 CN CN201811213854.2A patent/CN109006488A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004052085A1 (en) * | 2002-12-06 | 2004-06-24 | Del Monte Fresh Produce Company | Transgenic pineapple plants with modified carotenoid levels and methods of their production |
| CN101803570A (en) * | 2010-03-26 | 2010-08-18 | 浙江传化生物技术有限公司 | Method for efficiently propagating torch pineapples by utilizing in-vitro leaves |
| WO2013043508A1 (en) * | 2011-09-20 | 2013-03-28 | The Texas A&M University System | Citrus shoot regeneration compositions, methods, and systems |
| CN105028194A (en) * | 2015-06-23 | 2015-11-11 | 青岛农业大学 | Tissue culture rapid propagation method of tillandsia |
| CN106613940A (en) * | 2016-10-05 | 2017-05-10 | 南宁邃丛赋语科技开发有限责任公司 | In-vitro rapid propagation method for billbergia pyramidalis |
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| KISS E等: ""A novel method for rapid micropropagation of Pineapple"", 《HORTSCIENCE》 * |
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