CN107593449A - Induce the method that bud is formed in leaflet dragon bamboo test tube - Google Patents
Induce the method that bud is formed in leaflet dragon bamboo test tube Download PDFInfo
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- CN107593449A CN107593449A CN201711032751.1A CN201711032751A CN107593449A CN 107593449 A CN107593449 A CN 107593449A CN 201711032751 A CN201711032751 A CN 201711032751A CN 107593449 A CN107593449 A CN 107593449A
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Abstract
A kind of method for inducing and bud being formed in leaflet dragon bamboo test tube is provided, the sterile vegetative bud of growing thickly of leaflet dragon bamboo is randomly selected to be inoculated on Flower induction culture medium, every 21 days subcultures once, by 3 continuous Flower induction cultures and 1 renewal cultivation, had bud appearance at the 85th day or so.The leaflet dragon bamboo test tube Flower induction method of the present invention is simple to operate, it is time-consuming it is short, induced efficiency is high, reproducible, prediction will be bloomed to further investigate woody bamboo plant and florescence control provides research material, further manage the economic loss of bamboo grove to reduce scientific guidance is provided.
Description
Technical field：
The invention belongs to biological technical field, in particular it relates to which a kind of induce the side that bud is formed in leaflet dragon bamboo test tube
Method.
Background technology：
It is male that leaflet dragon bamboo (Dendrocalamus barbatus Hsueh et D.Z.Li) is under the jurisdiction of grass family Bambusoideae
Sinobambusa, it is large-scale Sympodial bamboos, is mainly distributed on the northern torrid zone of south of Yunnan, Vietnam, Burma and Laos and south subtropicses
Area.Leaflet dragon bamboo is a kind of excellent bamboo fibrid raw material, it has also become one of Economic house most with prospects.However,
Leaflet dragon bamboo Post flowering bamboo stalk is just withered, and bloomed even if indivedual bamboo clumps in bamboo grove in blocks also can cause full wafer bamboo grove to hold in several years
Death is spent, this brings about great losses to operator.Because bamboo large area is bloomed unobvious sign, and after death of blooming from
So renewal needs 8-10, and the practitioner to bamboo grove industry causes to perplex.
The foundation of bamboo test tube flowering experimental system is by for the physiological metabolism after understanding bamboo before flowering and gene table
The difference reached provides easy controllable experiment porch, so as to provide possibility for effective forecasting flower time, reduces and manages bamboo grove
Economic loss.
So far, there are no in the prior art be successfully established it is a kind of it is easy to operate, it is time-consuming it is short, induced efficiency is high, reproducible
Induction leaflet dragon bamboo test tube in form the report of bud.
The content of the invention：
It is contemplated that invention is a kind of to induce the method that bud is formed in leaflet dragon bamboo test tube, this method is simple to operate, consumption
When it is short, induced efficiency is high, reproducible, in 3 months can via leaflet dragon bamboo it is sterile grow thickly vegetative bud induction be spent
Bud.
In order to realize the above-mentioned purpose of the present invention, the invention provides following technical scheme：
It is explant to take leaflet dragon bamboo sterile test tube vegetative bud of growing thickly, every bottle connect 5 block bundle buds (3-5 buds/clump) carry out it is in vitro
Culture, cultivation temperature are 23 ± 2 DEG C, 40 μm of ol/m of intensity of illumination2S, light application time 12h/d.Minimal medium is MS, pH value
For 5.8, sucrose 3% is added, adds 5.6g/L agar solidified, 21 days are subculture cycle.First in MS+TDZ 1.0mg/L culture mediums
Continuous culture 3 times, then be transferred to culture medium MS+BA 2.0+NAA 0.2 and cultivate 1 time, culture 80-90 days to bud are grown.
Compared with prior art, the present invention possesses following excellent benefit：
The excellent benefit of the present invention is：Randomly select the sterile vegetative bud of growing thickly of leaflet dragon bamboo and be inoculated in Flower induction culture medium
On, every 21 days subcultures once, by 3 continuous Flower induction cultures and 1 renewal cultivation, in the 85th day or so existing bud
Occur.Leaflet dragon bamboo test tube Flower induction method that the present invention establishes is simple to operate, it is time-consuming it is short, induced efficiency is high, reproducible,
Prediction will be bloomed to further investigate woody bamboo plant and florescence control provides research material, be further to reduce to manage bamboo grove
Economic loss provides scientific guidance.
Brief description of the drawings
Fig. 1 is the growing state of clump bud during d group experiment process subcultures；
Fig. 2A is bud, and B is the gynoecium and stamen that dissection bud obtains.
Embodiment：
Further illustrate the essentiality content of the present invention with embodiments of the invention below, but this is not limited with this
Invention.
Embodiment 1：
1. sterile clonal foundation：
In 2009 in Dai Autonomous Prefecture of Xishuangbanna Mengla County collection leaflet dragon bamboo (Dendrocalamus
Barbatus) seed, the sterile clone of single seeded clump bud is built up by seed asepsis sprouting, and protected in the form of test tube clump bud
It is stored in Southwest China wildlife germplasm resource bank Vitro Plant storehouse.Concrete operations：Peel off collect leaflet dragon bamboo kind son
Exosper, with 75% alcohol surface sterilization 1min, then clean 2 times with distilled water, 5 minutes every time, then with 0.1% mercuric chloride
(HgCl2) solution disinfection 10min, sterile distilled water cleaning 5 times, it is enterprising that aseptic seed is then seeded in culture medium (1) 1/2MS
Row is sprouted.Minimal medium be MS (Murashige＆Skoog, 1962) (1/2MS is that MS culture medium a great number of elements halves, remaining
Components unchanged).
2nd, vegetative bud of growing thickly is bred：
The seedling that seed asepsis sprouting in step 1 obtains is forwarded to culture medium (2) MS+BA 2.0mg/L (unit is similarly hereinafter)
Clump bud propagation is carried out in+NAA 0.2, every 28 days subcultures once, obtain a large amount of vegetative buds of growing thickly, as follow-up after multiple subculture
The material of Flower induction.
3rd, the induction of bud and experiment process：
Flower induction culture medium：(3) MS+TDZ 1.0, (4) MS+TDZ 2.0；Recovery media：(2)MS+BA2.0+NAA
0.2.Above culture medium adds sucrose 3%, adds 5.6g/L agar solidified, Medium's PH Value 5.8.Condition of culture is：Temperature
23 ± 2 DEG C, 40 μm of ol/m of intensity of illumination2S, light application time 12h/d.The induction that five groups of experiments carry out bud is designed altogether：A. train
Support base (3) individually continuous squamous subculture；B. the independent continuous squamous subculture of culture medium (4)；C. continuously cultivated for 2 generations in (3) or (4)
It is transferred in (2) and cultivates again afterwards；D. culture in (2) is transferred to again after continuously cultivating for 3 generations in (3) or (4)；E. connect in (3) or (4)
It is transferred in (2) and cultivates again after 4 generations of continuous culture.Five groups of experiments, for a subculture cycle, observe the knot of subculture 6 times altogether with 21 days
Fruit.Each experiment process is at least inoculated with 10 bottles, and every bottle connects 5 block bundle buds (3-5 buds/clump), often handles 3 repetitions.It is real for every group
Test, since the 3rd subculture (the 43rd day), Multiple Buds were dissected every 3 days under Olympus SZ61 stereoscopes, until finding flower
Bud.
4th, the result of Flower induction：
Every group of experiment has the propagation of clump bud, and the proliferation rate of first time subculture is about 1：3-4, second of shoot proliferation rate
Increase, up to 1：5-6.Wherein, continuously cultivated in low concentration TDZ (1.0mg/L), vegetative bud of growing thickly can constantly increase
Grow, but clump bud becomes short and small, slowly albefaction.Culture more than 4 times in the high concentration TDZ (2.0mg/L), clump bud not only albefaction but also
Start dead.When being transferred in recovery media (2) continuous culture after continuously being cultivated 2-4 times in (3) or (4), nutrition of growing thickly
Bud continues to breed, and restoration ecosystem.Wherein, it is transferred to (2) after continuously cultivating 2 times, clump bud is grown tall and vanelets occurs, but is dissected not
See bud；It is transferred to after continuously cultivating 3 times, clump bud is grown tall in rosette-stape, and discovery bud is dissected after culture terminates in (2) for the first time
(see Fig. 1)；It is transferred to after continuously cultivating 4 times, albefaction clump bud slowly turns green and grown tall, but proliferation rate reduces, and Multiple Buds are extremely small and weak, pole
It is difficult to resolve and cuts open.Five groups of experiments the results are shown in Table 1.
The growing state of 1 five groups of experiment clump buds of table
5th, bud anatomical results：
Using the clump bud of d groups experiment induction as object, illustrate since the 3rd subculture (the 43rd day), in Olympus SZ61
Every 3 days results for dissecting vegetative bud of growing thickly under stereoscope.By being tracked dissection under stereoscope, opened from the 3rd subculture
Beginning terminates up to the 4th subculture, does not dissect bud, Multiple Buds now have been in rosette-stape fasciation.Continue to solve within 85th day
Cut open when growing thickly vegetative bud, discovery has bud appearance, continues to cultivate afterwards, bud ratio gradually increases.Terminate to the 6th subculture,
Anatomical results are shown, are clustered round in rosette-stape and vegetative bud is still mixed among bud together, and bud proportion is 70-90%,
Normal bud proportion is 60% or so, and lopsided bud is mainly manifested in stamen and gynoecium number is changed.Generally, just
Normal leaflet dragon bamboo stamen and gynoecium number are respectively 6 pieces 1 piece.The bud stamen number 6 pieces (60%) that group experiment obtains, 7
Piece (8.33%), 8 pieces (16.7%), 9 piece (8.33%) 10 pieces (8.33%), gynoecium have 2 pieces (8.33%) once in a while.By multiple
Squamous subculture, to normal bud, whole cycle is obtained it is about 85 days from nutrition vegetative bud of growing thickly.
To sum up experimental result, show that the best approach that bud is formed in induction leaflet dragon bamboo test tube is：Take leaflet dragon bamboo without
Bacterium test tube clump bud is explant, and every bottle connects 5 block bundle buds (3-5 buds/clump) and carries out cultured in vitro, and cultivation temperature is 23 ± 2 DEG C, illumination
40 μm of ol/m of intensity2S, light application time 12h/d.Minimal medium is MS, pH value 5.8, adds sucrose 3%, adds 5.6g/L
Agar solidified, every 21 days subcultures are once.It is first continuous in MS+TDZ 1.0mg/L culture mediums to cultivate 3 times, then it is transferred to culture medium MS+
BA 2.0+NAA 0.2 are cultivated 1 time, and at the 85th day or so, existing bud occurred., can be in 3 months via small based on the present invention
The sterile vegetative bud of growing thickly of leaf dragon bamboo obtains bud.
Claims (1)
1. induce the method that bud is formed in leaflet dragon bamboo test tube, it is characterised in that this method is to take leaflet dragon bamboo sterile test tube clump
Raw vegetative bud be explant, and every bottle connects 5 block bundle buds, is 3-5 buds/clump per block bundle bud, carries out cultured in vitro, cultivation temperature for 23 ±
2 DEG C, 40 μm of ol/m of intensity of illumination2S, light application time 12h/d, minimal medium MS, pH value 5.8, sucrose 3% is added,
Add 5.6g/L agar solidified, 21 days are subculture cycle, first continuous in MS+TDZ 1.0mg/L culture mediums to cultivate 3 times, then are transferred to
Culture medium MS+BA 2.0+NAA 0.2 are cultivated 80-90 days and grown to bud.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711032751.1A CN107593449B (en) | 2017-10-30 | 2017-10-30 | Method for inducing formation of flower buds in dendrocalamus parviflorus test tube |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711032751.1A CN107593449B (en) | 2017-10-30 | 2017-10-30 | Method for inducing formation of flower buds in dendrocalamus parviflorus test tube |
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| Publication Number | Publication Date |
|---|---|
| CN107593449A true CN107593449A (en) | 2018-01-19 |
| CN107593449B CN107593449B (en) | 2020-04-03 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201711032751.1A Active CN107593449B (en) | 2017-10-30 | 2017-10-30 | Method for inducing formation of flower buds in dendrocalamus parviflorus test tube |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116982559A (en) * | 2023-09-27 | 2023-11-03 | 中国科学院昆明植物研究所 | Primula forbesii test tube flowering induction medium and application thereof in test tube flowers |
-
2017
- 2017-10-30 CN CN201711032751.1A patent/CN107593449B/en active Active
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116982559A (en) * | 2023-09-27 | 2023-11-03 | 中国科学院昆明植物研究所 | Primula forbesii test tube flowering induction medium and application thereof in test tube flowers |
| CN116982559B (en) * | 2023-09-27 | 2023-12-29 | 中国科学院昆明植物研究所 | Test tube flowering induction culture method for primula sikkimensis |
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| CN107593449B (en) | 2020-04-03 |
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