CN104813943B - A kind of cultural method of hybrid cymbidium - Google Patents
A kind of cultural method of hybrid cymbidium Download PDFInfo
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- CN104813943B CN104813943B CN201510284906.5A CN201510284906A CN104813943B CN 104813943 B CN104813943 B CN 104813943B CN 201510284906 A CN201510284906 A CN 201510284906A CN 104813943 B CN104813943 B CN 104813943B
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Abstract
The invention discloses a kind of cultural method of hybrid cymbidium, it is therefore intended that and when solving at present using tissue culture technique breeding hybrid cymbidium, variation is inevitably produced, influences seedling quality, the problem of limiting protocorms reproduction speed.It comprises the following steps：Being attended to anything else using bud top, tissue induction protocorms, protocorms shoot proliferation, protocorms break up generation adventitious bud, adventitious bud strong seedling culture obtains seedling.The present invention can effectively solve the problem that the variation problem in hybrid cymbidium seeling industry, reduce the generation of variation, improve appreciation rate, at the same shorten the squamous subculture time more than half, in a disguised form improve the multiplication rates of protocorms.After measured, the present invention makes the proliferation rates of protocorms increase by 80 ~ 120%, and aberration rate reduces by 5 ~ 9%, efficiently solves the contradiction between hybrid cymbidium protocorms proliferation rates and aberration rate.Quick breeding and batch production large-scale production of the present invention for hybrid cymbidium seedling, has great importance, has preferable market prospects and application value.
Description
Technical field
The present invention relates to agriculture field, especially orchid cultivation field, specially a kind of cultural method of hybrid cymbidium.This
Invention can effectively solve the problem that the variation problem in hybrid cymbidium culture, have preferable effect.
Background technology
Hybrid cymbidium (Cymbidium grandifloruim) is that great Hua in Cymbidium (Cymbidium) grows nonparasitically upon another plant kind, and it has
There is an abundant gorgeous color, tall and big tall and straight spray, very large flower, the florescence of overlength, light delicate fragrance, noble makings,
It is deep to be liked by the people of other countries with high ornamental value and commodity value.
Hybrid cymbidium is transplanted extensively already on Japan, South Korea, the U.S., New Zealand, Europe, Taiwan and other places, and China great Hua
Orchid industry is also flourishing, and year, sales volume was million more than basin.With flourishing for hybrid cymbidium industry, traditional plant division is numerous
The needs of market can not be met by growing mode, have become hybrid cymbidium breeding through row industrial seedling rearing using tissue culture mode
Only way.
At present, in the industrial seedling rearing of hybrid cymbidium, mainly bred using tissue culture technique.Trained using tissue
The technology of supporting breeding hybrid cymbidium, can realize a large amount of, the quick breeding of seedling, and keep the character of kind.However, which
In incubation, variation is inevitably produced, influences seedling quality, or even job change character, meanwhile, variation also exists
The reproduction speed of protocorms is largely limited, loss is brought to the producer.
Therefore, there is an urgent need to a kind of new method, made a variation now with reducing or reducing hybrid cymbidium in tissue culture procedures
The generation of elephant.
The content of the invention
The goal of the invention of the present invention is：During for breeding hybrid cymbidium using tissue culture technique at present, unavoidably
Meeting produce variation, influence seedling quality, the problem of limiting protocorms reproduction speed, there is provided a kind of culture side of hybrid cymbidium
Method.The present invention can effectively solve the problem that the variation problem in hybrid cymbidium seeling industry, reduce the generation of variation, improve appreciation rate,
Shorten simultaneously the squamous subculture time more than half, in a disguised form improve the multiplication rates of protocorms.After measured, the present invention makes class former
The proliferation rates increase by 80~120% of bulb, aberration rate reduces by 5~9%, efficiently solves hybrid cymbidium protocorms increment speed
Contradiction between rate and aberration rate.Quick breeding and batch production large-scale production of the present invention for hybrid cymbidium seedling, has
Important meaning, there is preferable market prospects and application value.
To achieve these goals, the present invention adopts the following technical scheme that：
A kind of cultural method of hybrid cymbidium, comprises the following steps：
(1) protocorms are produced using hybrid cymbidium sprouting Stem tip induction；
(2) protocorms for inducing step 1 are inoculated in quiescent culture 14~16 days in liquid proliferated culture medium, obtain liquid
Body culture protocorms；
(3) the Liquid Culture protocorms for obtaining step 2 take out, and are inoculated with after cutting into solid multiplication medium culture 20
~30 days, obtain solid culture protocorms；
(4) quiescent culture in liquid proliferated culture medium will be inoculated in after solid culture protocorms cutting that step 3 obtains,
Step 2,3 are repeated in, after breeding is to required protocorms quantity, using solid multiplication medium culture 40~90 days, are obtained
To adventitious bud；
(5) adventitious bud for preparing step 4 is inoculated into Rooting and hardening-off culture base, is cultivated 90~120 days, is produced seedling；
In the step 2, condition of culture is：24~26 DEG C, 1500~2000LX of intensity of illumination of temperature, light application time is daily
15~18 hours, cultivate 14~16 days；
In the step 3, condition of culture is：24~26 DEG C, 1500~2000LX of intensity of illumination of temperature, light application time is daily
15~18 hours；
In the step 4, condition of culture is：24~26 DEG C, 1500~2000LX of intensity of illumination of temperature, light application time is daily
15~18 hours；
In the step 5, condition of culture is：24~26 DEG C, 2500~3000LX of intensity of illumination of temperature, light application time is daily
15~18 hours.
The constituent and concentration of the liquid proliferated culture medium be：500mg/L(NH4)2SO4, 250mg/L MgSO4·
7H2O, 525mg/L KNO3, 250mg/L KH2PO4, 200mg/L Ca3(PO4)2, 28mg/L Fe2(C4H4O6)3·2H2O,
7.5mg/L MnSO4·4H2O, 20000mg/L sugar；
The constituent and concentration of the solid multiplication culture medium be：500mg/L(NH4)2SO4, 250mg/L MgSO4·
7H2O, 250mg/L KH2PO4, 200mg/L Ca (NO3)2·4H2O, 28mg/L FeSO4·7H2O, 7.5mg/L MnSO4·
4H2O, 0.2mg/L 6-BA, 0.1mg/L NAA, 50000~100000mg/L banana, 4500mg/L agar, 20000mg/L sugar.
In the step 4, after breeding is to required protocorms quantity, using solid multiplication culture 60~90 days, obtain
Adventitious bud.
Adventitious bud prepared by step 4 is inoculated with cultivated 90~120 days in Rooting and hardening-off culture base after, obtain seedling, plant
Seedling can be transplanted into warmhouse booth plantation after hardening.The composition and concentration of the Rooting and hardening-off culture base be respectively：825mg/L
NH4NO3, 950mg/L KNO3, 220mg/L CaCl2·2H2O, 185mg/L MgSO4·7H2O, 85mg/L KH2PO4, 0.1mg/
L IBA, 100000mg/L bananas, 4500mg/L agar, 20000mg/L sugar.
In conventional tissue culture technology, protocorms easily produce variation, and aberration rate and increasing during shoot proliferation
It is directly proportional to be worth speed, therefore, the present invention provides a kind of cultural method of hybrid cymbidium, it mainly comprises the following steps：Utilize bud
Top is attended to anything else, and tissue induction protocorms, protocorms shoot proliferation, protocorms break up generation adventitious bud, adventitious bud strong sprout is trained
Support and obtain seedling.Compared with prior art, the present invention can significantly improve appreciation rate, reduce aberration rate.It has been found that using
For the method for the present invention in Liquid static culture, protocorms have broken growth polarity, and new protocorms are produced in spherical.
Applicant does contrast test using the common kind of multiple hybrid cymbidiums, the results showed that：Relative to prior art, this hair
Bright appreciation rate improves 80~120%, and aberration rate reduces by 5~9%.Meanwhile existing culture medium subculture once needs 30 days or so, and
The Liquid static culture subculture of the present invention once only needs 15~18 days, preferably 15 days, Subculture Time is effectively shortened, so as to become
The multiplication rate for improving protocorms of phase.In addition, banana is not added in the liquid proliferated culture medium of the present invention, so as to save
A part of production cost, the toxigenic capacity for reducing hybrid cymbidium, has great importance.
In summary, high-quality hybrid cymbidium seedling is cultivated using method of the invention, it is possible to quick, stable, for
Ensure seedling quality, promote the development of hybrid cymbidium, there is important progress meaning.
Embodiment
All features disclosed in this specification, or disclosed all methods or during the step of, except mutually exclusive
Feature and/or step beyond, can combine in any way.
Any feature disclosed in this specification, unless specifically stated otherwise, can be equivalent by other or with similar purpose
Alternative features are replaced.I.e., unless specifically stated otherwise, each feature is an example in a series of equivalent or similar characteristics
.
Embodiment 1
Fiery phoenix is one very popular kind of hybrid cymbidium, but it is during tissue culture, the propagation of protocorms
Speed is slow, and aberration rate is high, and effect of this technology in actual production is illustrated by taking this kind as an example.
Liquid proliferated culture medium is configured, the concentration of wherein each component is：500mg/L(NH4)2SO4、250mg/L MgSO4·
7H2O、525mg/L KNO3、250mg/L KH2PO4、200mg/L Ca3(PO4)2、28mg/L Fe2(C4H4O6)3·2H2O、
7.5mg/L MnSO4·4H2O, 20000mg/L sugar, the pH value of regulation liquid proliferated culture medium is 5.8, uses high pressure steam sterilization
Pot sterilizing 25 minutes or so, you can.
Solid multiplication culture medium is configured, the concentration of wherein each component is：500mg/L(NH4)2SO4、250mg/L MgSO4·
7H2O、250mg/L KH2PO4、200mg/L Ca(NO3)2·4H2O、28mg/L FeSO4·7H2O、7.5mg/L MnSO4·
4H2O, 0.2mg/L 6-BA, 0.1mg/L NAA, 50000~100000mg/L bananas, 4500mg/L agar, 20000mg/L sugar,
The pH value for adjusting solid multiplication culture medium is 5.8, is sterilized 25 minutes or so using high-pressure steam sterilizing pan, you can.
Rooting and hardening-off culture base is configured, the concentration of wherein each component is：825mg/L NH4NO3, 950mg/L KNO3,
220mg/L CaCl2·2H2O, 185mg/L MgSO4·7H2O, 85mg/L KH2PO4, 0.1mg/L IBA, 100000mg/L are fragrant
Any of several broadleaf plants, 4500mg/L agar, 20000mg/L sugar.
The incubation of fiery phoenix is as follows.
First, choose the protocorms of the fiery phoenix induced, more than diameter 0.3cm protocorms cutting into two,
It is inoculated in respectively in liquid proliferated culture medium and solid multiplication culture medium.Use Liquid Culture quiescent culture mode set code name as
A, this method are the method for present patent application；The mode code name of solid multiplication culture is set to B.The respectively inoculation 10 of two kinds of training methods
Bottle, 20 bulbs of every bottle of inoculation, and numbering is A1-A10, B1-B10 respectively.With the weight of every bottle of bulb of electronic balance scale.
A training methods：Liquid proliferated culture medium after inoculation is statically placed on culturing rack, culture room temperature maintains 25
DEG C, intensity of illumination 2000LX, daily 15 hours of light application time.After 15 days, the protocorms of cutting grow new class in incision
Protocorm, protocorms are distributed in peak green and radial.After new protocorms are continued into cutting, solid multiplication training is transferred to
Base culture is supported, the same Liquid Culture of condition of culture, new protocorms are grown after the 30th day.Again by new protocorms cutting, connect
Kind enters fluid nutrient medium quiescent culture 15 days, the culture of aforementioned liquids propagation base, solid multiplication base incubation is repeated, to the 60th
My god, terminate culture.
B training methods：Solid multiplication culture medium after inoculation is positioned on culturing rack, culture room temperature maintains 25
DEG C, intensity of illumination 2000LX, daily 15 hours of light application time.New protocorms are grown after 30 days.New protocorms are cut
After point, continue to be inoculated with into solid medium, continue culture 30 days.To the 60th day, terminate culture.
A, after two kinds of training methods of B cultivate 60 days, the situation of measurement protocorms propagation and variation.Measurement result is as follows
Shown in table 1, table 2.
The measurement result of the A training methods of table 1
| Numbering | Weight before culture | Weight after culture | Increase weight | Proliferation rate | Variation kind ball weight | Aberration rate |
| A1 | 3.911g | 13.656g | 9.745g | 249.2% | 1.598g | 11.7% |
| A2 | 3.845g | 13.258g | 9.413g | 244.8% | 0.993g | 7.5% |
| A3 | 4.135g | 15.145g | 11.010g | 266.3% | 1.327g | 8.8% |
| A4 | 3.523g | 14.025g | 10.502g | 298.1% | 1.654g | 11.8% |
| A5 | 4.552g | 14.987g | 10.435g | 229.2% | 1.025g | 6.8% |
| A6 | 3.014g | 13.125g | 10.111g | 335.5% | 0.765g | 5.8% |
| A7 | 3.985g | 14.156g | 10.171g | 255.2% | 1.145g | 8.1% |
| A8 | 3.651g | 14.268g | 10.617g | 290.8% | 1.695g | 11.9% |
| A9 | 4.256g | 15.079g | 10.823g | 254.3% | 1.753g | 11.6% |
| A10 | 4.856g | 15.675g | 10.819g | 222.8% | 1.114g | 7.1% |
| Average value | 3.973g | 14.337g | 10.364g | 260.9% | 1.307g | 9.1% |
The measurement result of the B training methods of table 2
| Numbering | Weight before culture | Weight after culture | Increase weight | Proliferation rate | Variation kind ball weight | Aberration rate |
| B1 | 4.581g | 10.225g | 5.644g | 123.2% | 1.520 | 14.9% |
| B2 | 3.176g | 9.647g | 6.471g | 203.7% | 1.325 | 13.7% |
| B3 | 3.259g | 9.364g | 6.105g | 187.3% | 1.844 | 19.7% |
| B4 | 3.478g | 9.977g | 6.499g | 186.9% | 1.536 | 15.4% |
| B5 | 5.002g | 10.372g | 5.370g | 107.4% | 1.379 | 13.3% |
| B6 | 3.921g | 9.191g | 5.270g | 134.4% | 1.628 | 17.7% |
| B7 | 4.868g | 9.358g | 4.490g | 92.2% | 1.723 | 18.4% |
| B8 | 3.074g | 8.949g | 5.875g | 191.1% | 1.963 | 21.9% |
| B9 | 3.368g | 9.652g | 6.257g | 185.8% | 1.364 | 14.1% |
| B10 | 3.951g | 9.455g | 5.504g | 139.3% | 1.337 | 14.1% |
| Average value | 3.867g | 9.619g | 5.452g | 141.0% | 1.562 | 16.2% |
After having measured, the protocorms that two kinds of training methods of A, B obtain all are inoculated in solid multiplication culture medium, 45 days
After grow adventitious bud, condition of culture temperature maintains 25 DEG C, intensity of illumination 2000LX, daily 15 hours of light application time.After 60 days
The bulb on adventitious bud is cut off, is inoculated into Rooting and hardening-off culture base.Condition of culture temperature maintains 25 DEG C, intensity of illumination
3000LX, daily 15 hours of light application time.After being cultivated 90 days in Rooting and hardening-off culture base, adventitious bud grows up to the strong of about 15cm length
Strong seedling.
Seedling is moved on into outdoor, bottle cap hardening is opened, is taken out after 1 day from bottle, and washes away root culture medium, is planted in temperature
In room.
After measured, the hybrid cymbidium seedling early growth of this example culture is healthy and strong, survival rate 100%.
Interpretation of result：By the as shown by data of table 1, table 2, A training method protocorms proliferation rates are higher by than B training method
119.9%, illustrate that the protocorms proliferation rate of A training methods is significantly higher than B training methods.A training methods protocorms make a variation
Rate lower than B training method 7.1%, illustrate that A training method protocorms aberration rates are substantially less than B training methods.
As a result show：The present invention can improve the appreciation rate of hybrid cymbidium protocorms, reduce its aberration rate, be that a kind of ratio is existing
In the conventional more preferable production method of hybrid cymbidium seeling industry.
The invention is not limited in foregoing embodiment.The present invention, which expands to, any in this manual to be disclosed
New feature or any new combination, and disclose any new method or process the step of or any new combination.
Claims (6)
1. a kind of cultural method of hybrid cymbidium, it is characterised in that comprise the following steps：
（1）Protocorms are produced using hybrid cymbidium sprouting Stem tip induction；
（2）The protocorms that step 1 is induced are inoculated in quiescent culture 14 ~ 16 days in liquid proliferated culture medium, obtain liquid training
Support protocorms；
（3）The Liquid Culture protocorms that step 2 is obtained take out, and are inoculated with after cutting into solid multiplication medium culture 20 ~ 30
My god, obtain solid culture protocorms；
（4）Quiescent culture in liquid proliferated culture medium will be inoculated in after solid culture protocorms cutting that step 3 obtains, successively
Repeat step 2,3, after breeding is to required protocorms quantity, using solid multiplication medium culture 40 ~ 90 days, it was indefinite to obtain
Bud；
（5）Adventitious bud prepared by step 4 is inoculated into Rooting and hardening-off culture base, is cultivated 90 ~ 120 days, is produced seedling；
In the step 2, condition of culture is：24 ~ 26 DEG C, 1500 ~ 2000LX of intensity of illumination of temperature, light application time daily 15 ~ 18
Hour, cultivate 14 ~ 16 days；
In the step 3, condition of culture is：24 ~ 26 DEG C, 1500 ~ 2000LX of intensity of illumination of temperature, light application time daily 15 ~ 18
Hour；
In the step 4, condition of culture is：24 ~ 26 DEG C, 1500 ~ 2000LX of intensity of illumination of temperature, light application time daily 15 ~ 18
Hour；
In the step 5, condition of culture is：24 ~ 26 DEG C, 2500 ~ 3000LX of intensity of illumination of temperature, light application time be daily 15 ~
18 hours；
The constituent and concentration of the liquid proliferated culture medium be：500mg/L (NH4)2SO4, 250mg/L MgSO4·7H2O,
525mg/L KNO3, 250mg/L KH2PO4, 200mg/L Ca3(PO4)2, 28mg/L Fe2(C4H4O6)3·2H2O, 7.5mg/L
MnSO4·4H2O, 20000mg/L sugar；
The constituent and concentration of the solid multiplication culture medium be：500mg/L (NH4)2SO4, 250mg/L MgSO4·7H2O,
250mg/L KH2PO4, 200mg/L Ca (NO3)2·4H2O, 28mg/L FeSO4·7H2O, 7.5mg/L MnSO4·4H2O,
0.2mg/L 6-BA, 0.1mg/L NAA, 50000 ~ 100000mg/L bananas, 4500mg/L agar, 20000mg/L sugar.
2. the cultural method of hybrid cymbidium according to claim 1, it is characterised in that in the step 2, condition of culture is：
25 DEG C, 1500 ~ 2000LX of intensity of illumination of temperature, daily 15 hours of light application time, cultivate 15 days.
3. the cultural method of hybrid cymbidium according to claim 1, it is characterised in that in the step 3,4, condition of culture
For：25 DEG C, 1500 ~ 2000LX of intensity of illumination of temperature, daily 15 hours of light application time.
4. the cultural method of hybrid cymbidium according to claim 1, it is characterised in that in the step 3, prepared by step 2
Liquid Culture protocorms take out, be inoculated with after cutting into solid multiplication culture medium quiescent culture 21 days, obtain solid culture class
Protocorm.
5. the cultural method of hybrid cymbidium according to claim 1, it is characterised in that in the step 4, when breeding is needed for
After protocorms quantity, using solid multiplication medium culture 60 ~ 90 days, adventitious bud is obtained.
6. the cultural method of hybrid cymbidium according to claim 1, it is characterised in that the adventitious bud for preparing step 4 is inoculated with
Enter after being cultivated 90 ~ 120 days in Rooting and hardening-off culture base, obtain seedling, seedling can be transplanted into warmhouse booth plantation after hardening.
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| CN109804927B (en) * | 2019-03-05 | 2020-06-05 | 湖北红杏生态环境科技有限公司 | In-vitro rapid propagation method of cymbidium hybridum |
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| US6020538A (en) * | 1998-05-01 | 2000-02-01 | Korea Kumho Petrochemical Co., Ltd. | Genetic transformation of orchids |
| CN1247022A (en) * | 1999-08-20 | 2000-03-15 | 华南师范大学 | Quickly in-vitro breeding method of macranthus Hui orchid |
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