CN109644872A - It is a kind of to hybridize the cultural method bloomed in sword-leaved cymbidium bottle - Google Patents
It is a kind of to hybridize the cultural method bloomed in sword-leaved cymbidium bottle Download PDFInfo
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- CN109644872A CN109644872A CN201910086076.3A CN201910086076A CN109644872A CN 109644872 A CN109644872 A CN 109644872A CN 201910086076 A CN201910086076 A CN 201910086076A CN 109644872 A CN109644872 A CN 109644872A
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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
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Abstract
The cultural method bloomed in a kind of hybridization sword-leaved cymbidium bottle, comprising the following steps: 1, choose big and full sword-leaved cymbidium hybridization fruit pod disinfection treatment；2, the fruit pod handled well incision is inoculated into initial culture base；3, the imperial root for choosing growing way health is forwarded to proliferated culture medium culture；4, differentiation seedling is gone to and continues to cultivate in root media；5, it goes in induced flowering culture medium and cultivates after 2~3 months, induced flowering culture medium are as follows: 1/2MS+NAA0.2mg/l+6-BA0.5mg/l+ spends No. 2 1~3g/L+KH of treasured2PO40.3~0.8g/L+ white granulated sugar 25g/L+ active carbon 0.5g/L+ peptone 1~3g/L+ carragheen, 7~8g/L.KH is added in induced flowering culture medium of the invention2PO4, largely promote the induction of bud, Flower induction rate is up to 80%~90%.
Description
Technical field
The present invention relates to technical field of plant culture, the cultural method specifically bloomed in a kind of hybridization sword-leaved cymbidium bottle.
Background technique
Sword-leaved cymbidium is ground plant, is born under sparse woods, in shrubbery, by mountain valley or in thick grass, 600-1800 meters of height above sea level.In originating in
State Anhui, Zhejiang, Jiangxi, Fujian, Taiwan, Hunan, Guangdong, Hainan, Guangxi, South-west Sichuan, Guizhou and Yunnan, it is widely distributed
In Southeast Asia and South Asia various countries, north to Japan.Breeding potential is extremely low under field conditions (factors) and blooms more difficult for orchid, therefore test tube is opened
Flower is also applied to more and more in orchid culture.
Application publication number is that the Chinese invention patent of CN 106171983A discloses a kind of culture side of sword-leaved cymbidium test tube flowering
Method, comprising the following steps: (1) material primary election: the sword-leaved cymbidium test tube seedling that selection no disease and pests harm grows fine, test tube seedling a height of 3~
6cm, root 3~6,2~3, leaf；(2) preculture: the sword-leaved cymbidium test tube seedling after selection is placed in initial culture base and is trained
Support, incubation time be 50~60d, cultivation temperature be 22~25 DEG C, light application time 14h/d, intensity of illumination be 1600~
2000lx, the initial culture base of replacement in every 10 days, the initial culture base are as follows: MS+1~3mg/L6-BA+0.1~0.2mg/L
NAA+28~32g/L sucrose+5~9g/L agar+0.1~0.2g/L active carbon；(3) Flower induction pre-processes: choosing preculture
Growth is consistent afterwards, growing way is healthy, the sword-leaved cymbidium seedling strain without insect pest pollution is placed in pre-culture medium and carries out preculture, and incubation time is
22~27d, cultivation temperature are 22~25 DEG C, light application time 14h/d, and intensity of illumination is 1600~2000lx, the preculture
Base are as follows: MS+2~4mg/L PP333+28~32g/L sucrose+5~9g/L agar+0.1~0.2g/L active carbon；(4) induction is opened
Flower: trimming Flower induction pretreated seedling strain root, be then placed in induced flowering culture medium and cultivated, and trains
Supporting the time is 100~120d, and cultivation temperature is 22~25 DEG C, light application time 14h/d, and intensity of illumination is 1600~2000lx,
The induced flowering culture medium of replacement in every 30 days, the induced flowering culture medium are as follows: MS+0.3~0.5mg/L TDZ+0.1~
0.2mg/L2,4-D+0.8~1.2mg/L ZT+28~32g/L sucrose+5~9g/L agar+0.1~0.2g/L active carbon.
The cultural method can promote the formation of bud, flower_bud formation is very fast by the combination between TDZ, 2,4-D and ZT
And bud is open normally, the florescence is long, grows fine, although test tube switch inductivity can be improved, its variability is big.And this is specially
Benefit is directly using test tube seedling as the culture materials of test tube flowering, and the test tube seedling screening time of excellent shape usually requires 5 months
The even longer time.
Summary of the invention
The present invention provides a kind of cultural method for hybridizing and blooming in sword-leaved cymbidium bottle, and main purpose is that existing sword-leaved cymbidium is overcome to try
Pipe bloom cultural method induced flowering variability is big, the defects of test tube seedling screening time is long.
The present invention adopts the following technical scheme:
It is a kind of to hybridize the cultural method bloomed in sword-leaved cymbidium bottle, comprising the following steps:
(1) material selection is with processing: it chooses big and full sword-leaved cymbidium and hybridizes fruit pod, handled using 0.1% mercuric chloride soaking disinfection, and
With in sodium thiosulfate and mercuric chloride raffinate；
(2) fruit pod is sowed: the fruit pod handled well being cut, is inoculated into initial culture base, and is placed on culturing room's culture, culturing room
Temperature is 22 ± 2 DEG C, and culture was to 4~6 months under half-light, the initial culture base are as follows: 1/2MS+NAA0.5~0.7mg/l+6-
BA0.1~0.2mg/l+ABT0.1~0.2mg/l+ white granulated sugar 25g/l+ active carbon 0.3~0.5g/L+ peptone 1~3g/L+ card
Draw 7~8g/L of glue；
(3) increment culture: the imperial root of growing way health is forwarded to proliferated culture medium and continues to cultivate in selecting step (2), culture
22 ± 2 DEG C of room temperature, intensity of illumination is 2000~2200lx, and light application time is 8~10h/d；
(4) culture of rootage: Multiplying culture is gone to after 3~4 months to be continued to cultivate in root media, cultivates 22 ± 2 DEG C of room temperature,
Intensity of illumination is 2000~2200lx, and light application time is 10~14h/d；
(5) induced flowering culture: culture of rootage goes in induced flowering culture medium after 2~3 months and cultivates 3~5 months, culturing room
22 ± 2 DEG C of temperature, intensity of illumination is 2000~2200lx, and light application time is 8~10h/d, the induced flowering culture medium are as follows: 1/
2MS+NAA0.2mg/l+6-BA0.5mg/l+ spends No. 2 1~3g/L+KH of treasured2PO40.3~0.8g/L+ white granulated sugar 25g/L+ active carbon
7~8g/L of 0.5g/L+ peptone 1~3g/L+ carragheen.
Specifically, the hybridization fruit pod that the sword-leaved cymbidium hybridization fruit pod is bright Yu Yujing Long Qidie.
Further, rise in value culture medium in the step (3) are as follows: 1/2MS+NAA0.2~0.5mg/l+6-BA3~5mg/l
7~8g/L of+white granulated sugar 30g/l+ active carbon 0.3g/L+ peptone 1~3g/L+ carragheen.
Further, root media in the step (4) are as follows: 1/2MS+NAA0.8~1mg/l+6-BA0~0.5mg/l
7~8g/L of+white granulated sugar 25g/L+ active carbon 0.5g/L+ peptone 1~3g/L+ carragheen.
Further, the initial culture base, the increment culture medium, the root media, induced flowering training
It supports and is added with riboflavin in base.
Further, the initial culture base, the increment culture medium, the root media and the induced flowering
The pH value of culture medium is adjusted to 5.4~5.6.
By the above-mentioned description of this invention it is found that the present invention is the advantages of hybridizing the cultural method bloomed in sword-leaved cymbidium bottle:
1, the present invention is directly sowed using hybrid seed and is cultivated in the medium, is then promoted it and is bloomed, can be in bottle directly
The separation situation for observing filial generation character, substantially reduces the screening time of merit.And it directlys adopt seed and broadcasts
Kind tissue culture, operability is stronger, substantially reduces cultivation screening time.
2, KH is added in induced flowering culture medium of the invention2PO4, wherein the concentration of P ion is 0.3~0.8g/L, can
Largely promote the induction of bud, Flower induction rate is up to 80%~90%.
3, all culture mediums of the invention do not add the additives such as banana, potato and coconut milk, but dense with peptone
1~3g/L is spent to replace, and is reduced the workload of culture medium preparation, is reduced culture medium cost, while decreasing pollution rate.
Detailed description of the invention
Fig. 1 is the figure of blooming of processing F in induced flowering culture of the present invention.
Specific embodiment
It elaborates with reference to embodiments to the present invention.
The cultural method bloomed in hybridization sword-leaved cymbidium bottle of the invention, comprising the following steps:
1, material selection is with processing: choosing big and full sword-leaved cymbidium and hybridizes fruit pod, is handled, be used in combination using 0.1% mercuric chloride soaking disinfection
In sodium thiosulfate and mercuric chloride raffinate.Wherein, the hybridization fruit pod that sword-leaved cymbidium hybridization fruit pod is bright Yu Yujing Long Qidie.
2, fruit pod is sowed: the fruit pod handled well being cut, is inoculated into initial culture base, and is placed on culturing room's culture, institute
State initial culture base are as follows: 1/2MS+NAA0.5~0.7mg/l+6-BA0.1~0.2mg/l+ABT0.1~0.2mg/l+ white granulated sugar
25g/l+ active carbon 0.3~0.5g/L+ peptone 1~3g/L+, 7~8g/L of carragheen, pH value 5.4~5.6.
Condition of culture: culture room temperature is 22 ± 2 DEG C, is cultivated 4~6 months under half-light.
The influence that the different initial culture bases of table 1 sprout fruit pod
| Processing number | Culture medium | It sows number (bottle) | Sprout time (day) |
| 1 | 1/2MS+NAA0.5mg/L+6BA0.1mg/L+ABT0.1mg/L+ white granulated sugar 25g/l+ active carbon 0.3g/L+ peptone 1g/L+ carragheen 7g/L | 20 | 110 |
| 2 | 1/2MS+NAA0.5mg/L+6BA0.2mg/L+ABT0.1mg/L+ white granulated sugar 25g/l+ active carbon 0.4g/L+ peptone 2g/L+ carragheen 7.5g/L | 20 | 112 |
| 3 | 1/2MS+NAA0.6mg/L+6-BA0.1mg/L+ABT0.1mg/L+ white granulated sugar 25g/l+ active carbon 0.4g/L+ peptone 2g/L+ carragheen 7g/L | 20 | 120 |
| 4 | 1/2MS+NAA0.6mg/L+6-BA0.2mg/L+ABT0.2mg/L+ white granulated sugar 25g/l+ active carbon 0.4g/L+ peptone 2g/L+ carragheen 7g/L | 20 | 100 |
| 5 | 1/2MS+NAA0.7mg/L+6-BA0.1mg/L+ABT0.2mg/L+ white granulated sugar 25g/l+ active carbon 0.5g/L+ peptone 3g/L+ carragheen 8g/L | 20 | 108 |
| 6 | 1/2MS+NAA0.7mg/L+6-BA0.2mg/L+ABT0.2mg/L+ white granulated sugar 25g/l+ active carbon 0.5g/L+ peptone 3g/L+ carragheen 8g/L | 20 | 110 |
| 7 | 1/2MS+NAA0.5mg/L+6-BA0.1mg/L+ white granulated sugar 25g/l+ active carbon 0.3g/L+ peptone 1g/L+ carragheen 7g/L | 20 | 150 |
| 8 | 1/2MS+NAA0.6mg/L+6-BA0.2mg/L+ white granulated sugar 25g/l+ active carbon 0.4g/L+ peptone 2g/L+ carragheen 7g/L | 20 | 165 |
| 9 | 1/2MS+NAA0.7mg/L+6-BA0.1mg/L+ white granulated sugar 25g/l+ active carbon 0.5g/L+ peptone 3g/L+ carragheen 8g/L | 20 | 160 |
From the point of view of table 1, sword-leaved cymbidium fruit pod is relatively early sprouted on the culture medium of processing numbers 4, and when observation in 100 days just has very beautiful
Long Gen in processing number 7,8,9 relatively late wants five wheat harvesting periods or so, and other upper sword-leaved cymbidium fruit pods of processing number for having addition ABT
Sprouting time differ with processing numbers 4 be not very greatly.It can thus be seen that ABT still have to the sprouting of sword-leaved cymbidium fruit pod it is biggish
It influences, it can promote the sprouting of seed earlier in sword-leaved cymbidium fruit pod.
3, increment culture
The imperial root of growing way health is forwarded to proliferated culture medium and continues to cultivate in selecting step (2).Rise in value culture medium are as follows: 1/
2MS+NAA0.2~0.5mg/l+6-BA3~5mg/l+ white granulated sugar 30g/l+ active carbon 0.3g/L+ peptone 1~3g/L+ OK a karaoke club
7~8g/L of glue.
Condition of culture: 22 ± 2 DEG C of room temperature of culture, intensity of illumination are 2000~2200lx, and light application time is 8~10h/d,
Light source is fluorescent lamp, is cultivated 3~4 months.
4, culture of rootage
Multiplying culture is gone to after 3~4 months to be continued to cultivate in root media.Root media are as follows: 1/2MS+NAA0.8~
7~8g/L of 1mg/l+6-BA0~0.5mg/l+ white granulated sugar 25g/L+ active carbon 0.5g/L+ peptone 1~3g/L+ carragheen.
Condition of culture: 22 ± 2 DEG C of room temperature of culture, intensity of illumination are 2000~2200lx, and light application time is 10~14h/
D, light source are fluorescent lamp, are cultivated 2~3 months.
5, induced flowering culture: culture of rootage goes in induced flowering culture medium after 2~3 months and cultivates 3~5 months, institute
State induced flowering culture medium are as follows: 1/2MS+NAA0.2mg/l+6-BA0.5mg/l+ spends No. 2 1~3g/L+KH of treasured2PO40.3~
7~8g/L of 0.8g/L+ white granulated sugar 25g/L+ active carbon 0.5g/L+ peptone 1~3g/L+ carragheen.
Condition of culture: 22 ± 2 DEG C of room temperature of culture, intensity of illumination are 2000~2200lx, and light application time is 8~10h/d,
Light source is fluorescent lamp, is cultivated 3~5 months.
Influence of the induced flowering culture medium of 2 heterogeneity of table and various concentration to cultivating seedling induced flowering
| Processing number | Culture medium | Inoculation number (strain) | Flower bud number (a) | Flower induction rate (%) |
| A | 1/2MS+NAA0.2mg/L+6-BA0.5mg/L+ spends No. 2 1g/L+ white granulated sugar 25g/L+ active carbon 0.5g/L+ peptone 1g/L+ carragheen 7g/L of treasured | 50 | 25 | 50 |
| B | 1/2MS+NAA0.2mg/L+6-BA0.5mg/LL+ spends No. 2 3g/L+ white granulated sugar 25g/L+ active carbon 0.5g/L+ peptone 3g/L+ carragheen 8g/L of treasured | 50 | 30 | 60 |
| C | 1/2MS+NAA0.2mg/L+6-BA0.5mg/L+ spends No. 2 1g/L+KH of treasured2PO40.3g/L+ white granulated sugar 25g/L+ active carbon 0.5g/L+ peptone 1g/L+ carragheen 7g/L | 50 | 40 | 80 |
| D | 1/2MS+NAA0.2mg/L+6-BA0.5mg/L+ spends No. 2 1g/L+KH of treasured2PO40.5g/L+ white granulated sugar 25g/L+ active carbon 0.5g/L+ peptone 1g/L+ carragheen 7g/L | 50 | 42 | 84 |
| E | 1/2MS+NAA0.2mg/L+6-BA0.5mg/L+ spends No. 2 1g/L+KH of treasured2PO40.8g/L+ white granulated sugar 25g/L+ active carbon 0.5g/L+ peptone 2g/L+ carragheen 7g/L | 50 | 43 | 86 |
| F | 1/2MS+NAA0.2mg/L+6-BA0.5mg/L+ spends No. 2 2g/L+KH of treasured2PO40.5g/L+ white granulated sugar 25g/L+ active carbon 0.5g/L+ peptone 3g/L+ carragheen 8g/L | 50 | 45 | 90 |
| G | 1/2MS+NAA0.2mg/L+6-BA0.5mg/L+ spends No. 2 3g/L+KH of treasured2PO40.5g/L+ white granulated sugar 25g/L+ active carbon 0.5g/L+ peptone 3g/L+ carragheen 8g/L | 50 | 43 | 86 |
| H | 1/2MS+NAA0.2mg/L+6-BA0.5mg/L+ spends No. 2 2g/L+KH of treasured2PO40.8g/L+ white granulated sugar 25g/L+ active carbon 0.5g/L+ peptone 3g/L+ carragheen 8g/L | 50 | 44 | 88 |
| I | 1/2MS+NAA0.2mg/L+6-BA0.5mg/L+ spends No. 2 3g/L+KH of treasured2PO40.8g/L+ white granulated sugar 25g/L+ active carbon 0.5g/L+ peptone 3g/L+ carragheen 8g/L | 50 | 42 | 84 |
From table 2 it can be seen that processing A, B do not add KH2PO4, Flower induction rate is significant lower；Processing C has added KH2PO4
Flower induction rate significantly improves, and while processing F is proliferated, its Flower induction rate reaches peak 90%(such as Fig. 1).Thus
As can be seen that adding KH in culture medium2PO4, the concentration of P ion is 0.3~0.8g/L, largely promotes luring for bud
It leads, Flower induction rate is up to 80%~90%.
A little riboflavin is added in the culture medium in above each stage, avoids medium browning.The PH of each stage culture medium
Value is adjusted to 5.6 or so.
The above is only a specific embodiment of the present invention, but the design concept of the present invention is not limited to this, all to utilize this
Design makes a non-material change to the present invention, and should all belong to behavior that violates the scope of protection of the present invention.
Claims (6)
1. the cultural method bloomed in a kind of hybridization sword-leaved cymbidium bottle, which comprises the following steps:
(1) material selection is with processing: it chooses big and full sword-leaved cymbidium and hybridizes fruit pod, handled using 0.1% mercuric chloride soaking disinfection, and
With in sodium thiosulfate and mercuric chloride raffinate；
(2) fruit pod is sowed: the fruit pod handled well being cut, is inoculated into initial culture base, and is placed on culturing room's culture, culturing room
Temperature is 22 ± 2 DEG C, and culture was to 4~6 months under half-light, the initial culture base are as follows: 1/2MS+NAA0.5~0.7mg/l+6-
BA0.1~0.2mg/l+ABT0.1~0.2mg/l+ white granulated sugar 25g/l+ active carbon 0.3~0.5g/L+ peptone 1~3g/L+ card
Draw 7~8g/L of glue；
(3) increment culture: the imperial root of growing way health is forwarded to proliferated culture medium and continues to cultivate in selecting step (2), culture
22 ± 2 DEG C of room temperature, intensity of illumination is 2000~2200lx, and light application time is 8~10h/d；
(4) culture of rootage: Multiplying culture is gone to after 3~4 months to be continued to cultivate in root media, cultivates 22 ± 2 DEG C of room temperature,
Intensity of illumination is 2000~2200lx, and light application time is 10~14h/d；
(5) induced flowering culture: culture of rootage goes in induced flowering culture medium after 2~3 months and cultivates 3~5 months, culturing room
22 ± 2 DEG C of temperature, intensity of illumination is 2000~2200lx, and light application time is 8~10h/d, the induced flowering culture medium are as follows: 1/
2MS+NAA0.2mg/l+6-BA0.5mg/l+ spends No. 2 1~3g/L+KH of treasured2PO40.3~0.8g/L+ white granulated sugar 25g/L+ active carbon
7~8g/L of 0.5g/L+ peptone 1~3g/L+ carragheen.
2. the cultural method bloomed in a kind of hybridization sword-leaved cymbidium bottle as described in claim 1, it is characterised in that: the sword-leaved cymbidium hybridization
Fruit pod is the hybridization fruit pod of bright Yu Yujing Long Qidie.
3. the cultural method bloomed in a kind of hybridization sword-leaved cymbidium bottle as described in claim 1, it is characterised in that: the step (3)
Middle increment culture medium are as follows: 1/2MS+NAA0.2~0.5mg/l+6-BA3~5mg/l+ white granulated sugar 30g/l+ active carbon 0.3g/L+ egg
7~8g/L of white 1~3g/L+ of peptone carragheen.
4. the cultural method bloomed in a kind of hybridization sword-leaved cymbidium bottle as described in claim 1, it is characterised in that: the step (4)
Middle root media are as follows: 1/2MS+NAA0.8~1mg/l+6-BA0~0.5mg/l+ white granulated sugar 25g/L+ active carbon 0.5g/L+ egg
7~8g/L of white 1~3g/L+ of peptone carragheen.
5. the cultural method bloomed in a kind of hybridization sword-leaved cymbidium bottle as described in claim 1, it is characterised in that: the Initial culture
Riboflavin is added in base, the increment culture medium, the root media and the induced flowering culture medium.
6. the cultural method bloomed in a kind of hybridization sword-leaved cymbidium bottle as described in claim 1, it is characterised in that: the Initial culture
Base, the pH value for rising in value culture medium, the root media and the induced flowering culture medium are adjusted to 5.4~5.6.
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| CN111642393B (en) * | 2020-04-30 | 2023-08-29 | 广西壮族自治区农业科学院 | Tissue culture method for inducing flowering in cymbidium bottle |
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