CN107232061A - A kind of special culture media and tissue culture method of rapid induction bletilla bulb - Google Patents
A kind of special culture media and tissue culture method of rapid induction bletilla bulb Download PDFInfo
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- CN107232061A CN107232061A CN201710552200.1A CN201710552200A CN107232061A CN 107232061 A CN107232061 A CN 107232061A CN 201710552200 A CN201710552200 A CN 201710552200A CN 107232061 A CN107232061 A CN 107232061A
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- bletilla
- bulb
- culture
- seed
- pseudobulb
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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
Abstract
The invention discloses a kind of special culture media of rapid induction bletilla bulb, the special culture media by bletilla seed germination medium and bulb are efficiently induced, strong seedling culture base is constituted, wherein：The formula of the bletilla seed germination medium is：1/2MS, NAA0.1mg/L, GA30.1mg/L, the 7.0g/L of agar 6.5, white sugar 20g/L, pH5.8 6.0；It is described and bulb is efficiently induced, the formula of strong seedling culture base is：1/2MS, NAA0.5~0.6mg/L, 6 BA0.6~0.8mg/L, PP3330.4~0.5mg/L, white sugar 30 45g/L, 25~40g/L of banana puree, activated carbon 0.1g/L, pH5.8~6.0.The invention also discloses the tissue culture method using the special culture media.This method simplifies the production link of bletilla pseudobulb tissue-cultured seedling, induce pseudobulb, pseudobulb fast-growth, and Rooting and hardening-off culture is completed in same step, it it is 90 days in from seed to required time is transplanted, shorten incubation time, pseudobulb differentiation rate, the fast-growth of pseudobulb are promoted, the transplanting survival rate of tissue-cultured seedling is improved.
Description
Technical field
The invention belongs to bletilla technical field of cultivation, and in particular to a kind of special culture media of rapid induction bletilla bulb and
Tissue culture method.
Background technology
Bletilla is that (Bletilla striata (Thunb.) Reichb.f., its pseudobulb is medicinal portion to orchid bletilla
Position, has effects that astringing to arrest bleeding, detumescence and promoting granulation, and now research finds bletilla effect in terms for the treatment of pulmonary tuberculosis, stomach trouble, tumour
Substantially, while the still chemical material of high-quality.But bletilla plant is destroyed by artificial excavation, raw mirror, and wild resource is very rare,
It has been listed in rare plant second class protection kind.
With the development need of bletilla value of exploiting and utilizing, its market demand is continuously increased, and wild resource ten
Divide breeding potential under rareness, bletilla natural conditions relatively low, so be badly in need of carrying out bletilla artificial propagation and the research of standardized cultivation,
So as to preferably protect bletilla wild resource, and ensure the market demand and the exploitation of bletilla.
Under the conditions of traditional cultivation, bletilla is based on division propagation, and its reproduction speed is slow, and breeding coefficient is extremely low, and easily leads
The accumulation and propagation for evil of causing a disease, it is difficult to the need for suitable for modern is planted.And seed amount is big in bletilla Fruit pod, but because seed knot
Structure is special, is difficult to sprout under natural conditions, so the rare bottleneck as its large-scale development of bletilla seedling.And with biological skill
The development of art, bletilla tissue culture technology has obtained very fast development, typically all passes through aseptic seeding, tufted seedling culture, strong seedling culture, life
Multiple links such as root culture, and it is work consuming, time-consuming, and tissue-cultured seedling bulb differentiation rate is not high under normal circumstances, the transplant survival of tissue-cultured seedling
Rate is not also high, so as to largely have impact on the production application of bletilla tissue-cultured seedling.So how to simplify tissue culture link, reduce into
This, promotes the differentiation of bulb, fast-growth to turn into the key factor of influence tissue-cultured seedling transplanting survival rate and production application.
201110119312.0 also disclose that a kind of method that bletilla pseudobulb is prepared in culture vessel and its special culture
Base, which describes the special culture media using bletilla seed as explant, by seed germination medium, pseudobulb inducing culture,
Pseudobulb proliferated culture medium and pseudobulb root media composition.Each of which step needs a specific culture medium, and it is trained
Foster mode has the defect such as work consuming, time-consuming as described above.
The content of the invention
For the defect of existing bletilla bulb tissue culture technology, it is an object of the invention to provide a kind of rapid induction bletilla squama
The special culture media and tissue culture method of stem, methods described are simple to operate, and production cost is low, and effect stability.
To reach above-mentioned purpose, the present invention provides following technical scheme：
1st, a kind of special culture media of rapid induction bletilla bulb, the special culture media is by bletilla seed germination medium
Efficiently induced with bulb and strong seedling culture base composition, wherein：
The formula of the bletilla seed germination medium is：1/2MS, NAA0.1mg/L, GA30.1mg/L, agar 6.5~
7.0g/L, white sugar 20g/L, pH5.8~6.0；Described and bulb is efficiently induced and the formula of strong seedling culture base is：1/2MS,
NAA0.5~0.6mg/L, 6-BA0.6~0.8mg/L, PP3330.4~0.5mg/L, 30~45g/L of white sugar, banana puree 25~
40g/L, activated carbon 0.1g/L, pH5.8-6.0.
It is preferred that, the bulb is efficiently induced and the formula of strong seedling culture base is：1/2MS, NAA0.6mg/L, 6-
BA0.8mg/L, PP3330.5mg/L, white sugar 40g/L, banana puree 25g/L, activated carbon 0.1g/L, pH5.8-6.0.
2nd, a kind of tissue culture method of rapid induction bletilla bulb, it is characterised in that comprise the following steps：
1) sterilization of bletilla seed：Uncracked bletilla seed mass fraction is washed into surface for 75% alcohol painting
Fruit pod, is then placed in 0.1% mercuric chloride 15~20min of immersion by 1-3min on superclean bench, then with aseptic water washing 3
~5 times, aseptic filter paper blots surface moisture in case inoculation；
2) under sterile condition of work, by step 1) the bletilla seed of sterilization is inoculated into kind described in claim 1
Cultivated in sub- germination medium；
3) when step 2) bulb that is forwarded to described in claim 1 when sprouting 0.8~1.2cm of height of seedling of the bletilla seed is high
Cultivated in effect induction and strong seedling culture base.
It is further preferred that step 2) condition of culture be：Illumination 1500~2000Lx, 10~12h/ days；Temperature, 24~
28℃。
It is further preferred that step 3) condition of culture be：Illumination 1500~2000Lx, 12~15h/ days；Temperature, 24~
28℃。
Further, the collection of bletilla seed described in the above method and storage practice are：Gather 9-10 month bletillas ripe, full
Full Fruit pod is placed in the place of shady and cool ventilation, spreading for cooling 2~3 weeks；When Fruit pod epidermis is changed into light brown, Fruit pod is loaded on kraft paper bag
In be stored in 4 DEG C of refrigerating chambers of refrigerator, it is ensured that used in bletilla tissue-cultured seedling whole year production.
NAA used is methyl α-naphthyl acetate, GA in the present invention3For gibberellin, 6-BA is 6- benzyl aminoadenines, PP333For multiple-effect
Azoles, reagent is that analysis is pure, and water is ultra-pure water.
The beneficial effects of the present invention are：Then this method is used using promoting seed quick-speed germination as first stage culture
Efficient culture medium, i.e., to promote the hormone 6- benzyl aminoadenines quickly broken up, adjust Furcation defects, the methyl α-naphthyl acetate of growth, and
Adjust plant hormone balance, improve the paclobutrazol of resistance, three kinds of hormones are formula, screen suitable concentration and ratio, make it
The preferably false seedling bulb of regulation and control bletilla tissue culture, cauline leaf, the differentiation and growth of root；And add the suitable nutrition original-white sugar of concentration and
Banana puree, has been effectively promoted the fast-growth of bulb, and appropriate activated carbon can preferably promote the growth of root, and can be effective
Some toxin produced in absorption bletilla growth.So, the cultural method simplifies the production link of bletilla pseudobulb tissue-cultured seedling,
Collect induction pseudobulb, pseudobulb fast-growth and Rooting and hardening-off culture to complete in a step, be 90 from seed to required time is transplanted
In it, incubation time is shortened, pseudobulb differentiation rate is improved, the fast-growth of pseudobulb is promoted, improves tissue-cultured seedling
Transplanting survival rate.
Brief description of the drawings
In order that the purpose of the present invention, technical scheme and beneficial effect are clearer, the present invention provides drawings described below and carried out
Explanation：
Fig. 1 is the diameter test chart of one plant of bletilla pseudobulb；
The plant height test chart of Fig. 2 bletilla pseudobulbs；
The plant height test chart of Fig. 3 bletilla pseudobulbs；
Fig. 4 is the bletilla pseudobulb growing state after culture dish culture 60 days；
Fig. 5 is the growing state after transplanting 20 days.
Embodiment
Below in conjunction with accompanying drawing, the preferred embodiments of the present invention are described in detail.
Following examples use ripe uncracked bletilla capsule and (adopted in Chongqing City's medicine plantation research institute experiment base
Ground), Fruit pod store method, micro- yellow Fruit pod is gathered in the 8-10 months, and ventilation is dried in the air 5-10 days indoors, treats that Fruit pod epidermis is changed into shallow
During brown, it is put into kraft envelope, is placed in 4 DEG C of fridge freshness retaining rooms, this method can effectively preserves seed vitality and be easy to week
The production in year.Culture medium autoclaving 17min under the conditions of 121 DEG C, it is standby.
Seed Fruit pod used is sterilized and is inoculated with the following manner in following examples：Uncracked Fruit pod flowing water is washed
Only, applied with 75% alcohol and wash 1~3min of surface, it is 0.1% that Fruit pod then is placed in into mass fraction on superclean bench
15~20min is soaked in mercuric chloride, then with aseptic water washing 3-5 time, aseptic filter paper blots surface moisture in case inoculation, is being inoculated with
The Fruit pod sterilized is cut off into osculum with sterile scissors in disk, dipping seed with aseptic inoculation ring is uniformly coated in germination medium
On.
The screening of bletilla axenic germination culture medium
According to the formula design shown in table 1, bletilla seed asepsis sprouting is observed, condition of culture is：Illumination
1500--2000Lx, 10-12h/ days；Temperature, 24--28 DEG C.Bletilla sprouting height of seedling, root hair situation are counted at 4 weeks as shown in table 2：
Table 1：The formula design of bletilla aseptic culture medium
Table 2：Bletilla seed germination and growth situation on different germination mediums
From table 2 it can be seen that in condition of culture, 1/2MS is good compared with MS culture effects under the same conditions, and several culture mediums
Contrast finds that (4) culture medium prescription is with the obvious advantage in terms of startup, seedling growth is sprouted, so more suitable bletilla is quickly sprouted
Hair needs, and preferably germination medium formula is：1/2MS+NAA0.1mg/L+GA30.1mg/L+ white sugar 20g/L.
Bulb efficiently induction and the screening of strong seedling culture base
High 0.8~1.2cm bulb is transferred to table 3 and enumerates progress switching culture, condition of culture in shown culture medium
For pH5.8-6.0；Illumination 1500--2000Lx, 12-15h/ days；Temperature, 24--28 DEG C；To the bulb growth to each culture medium
Situation is observed and counted, and is recorded in table 4.
Efficiently induction and the design of strong seedling culture based formulas of the bulb of table 3
The bulb of table 4 is efficiently induced, strong seedling culture base screening situation
As seen from Table 4, A formulas root growth is prosperous, have impact on the differentiation and growth of bulb；H culturing gene paclobutrazol concentration
It is higher, there is point evil substantially, and bulb, plant strain growth are all suppressed；D, E, F are formulated, and root growth is good, but bulb differentiation and
Growing way is poor, breaks up more very thin seedling；B, C, G are formulated, because of 6-BA, PP333Concentration and match more suitable, its bulb
Differentiation and growing way are preferable, wherein the most pronounced effects in terms of bulb induces differentiation, bulb growth, strong sprout are formulated with C, so suitably
Bletilla bulb is efficiently induced, the formula of strong seedling culture is 1/2MS, NAA0.6mg/L, 6-BA0.8mg/L, PP3330.5mg/L, in vain
Sugared 40g/L, banana puree 25g/L, activated carbon 0.1g/L.
The tissue-cultured seedling (switching 60 days) that further table 4 is cultivated is transplanted, and corkage hardening 1 week is transplanted to covering
On fertile soil 3-5cm ridge, and note sheltering from heat or light and moisturizing, statistics survival rate and growing way, are recorded in shown in table 5 when transplanting 20 days：
Table 5：The survival rate statistics of transplanted seedling
" _ ", represents long potential difference, and "+" represents that growing way is preferable.
As can be seen from Table 5, the transplanted seedling growth potential and situation of taking root of C formulas are preferable.
The bletilla bulb rapid induction tissue culture method of embodiment 1
Step 1：Fruit pod requirement, using light brown, ripe bletilla Fruit pod is become, the Fruit pod of routine preservation is easy to infection miscellaneous bacteria
And rupture, it is impossible to meet needed for bletilla tissue-cultured seedling whole year production, it is 4 DEG C of preservations to be encased using brown paper in refrigerator cold-storage；
Step 2：Uncracked Fruit pod is applied with 70% alcohol and washes surface 1-3min, then will fruit on superclean bench
Pod, which is placed in the mercuric chloride that mass fraction is 0.1%, soaks 15--20min, then with aseptic water washing 3--5 times, aseptic filter paper is blotted
Surface moisture is in case inoculation；On superclean bench, the Fruit pod sterilized is cut off into osculum with sterile scissors, aseptic inoculation ring is used
Dip seed to be uniformly coated on the culture medium of high-temperature sterilization, culture medium prescription is 1/2MS+NAA0.1mg/L+GA30.1mg/L+
Agar powder 6.5-7.0g/L+ white sugar 20g/L, pH5.8-6.0；Condition of culture is that condition of culture is illumination 1500--2000Lx,
10-12h/ days；Temperature, is cultivated 4 weeks or so by 24--28 DEG C, obtains band 1-2 piece leaflets, plant height 0.8-1.2cm seedling；
Step 3：Carry out switching culture, culture medium prescription 1/2MS+NAA0.6mg/L+6-BA0.8mg/L+PP3330.5mg/L
+ white sugar 40g/L+ banana puree 25g/L+ activated carbons 0.1g/L, pH5.8-6.0, sterilized rear use；The culture medium is bletilla pseudobulb
Induction, fast-growth, the strong seedling culture base taken root；Condition of culture：Illumination 1500--2000Lx, 12-15h/ days；Temperature, 24--
28℃；On switching culture medium, pseudobulb induction and fast-growth observation：Seedling transfer on pseudobulb efficient culture medium, passes through
Cultivate within 2-3 weeks, small pseudobulb has broken up, counted at 4-5 weeks, bulb differentiation rate is more than 90%, and pseudobulb growth is fast
Speed；During by 8 weeks, pseudobulb diameter be more than 0.4cm for more than 90%, plant height 6-10cm, coring is flourishing；Fig. 1 is one plant of bletilla
The diameter test chart of pseudobulb, as can be seen from Figure 1 its diameter reach 9.41mm, Fig. 2, Fig. 3 are the plant height of two plants of bletilla pseudobulbs
Test chart, Fig. 4 is the bletilla pseudobulb growing state after culture dish culture 60 days.
Step 4：Hardening, transplanting, cultivating seedling is moved on to from culturing room and hardening is opened under normal temperature environment 1 week, is cleaned, field planting
Onto the case face for transplanting seedlings ground, root water is watered, and with sunshade rate 40-60% sunshade net sunshade, per watering within 3-5 days 1 time, keep
Humidity.30 days or so, new root and young leaves were grown, and counted survival rate, and Fig. 5 is the growing state after transplanting 20 days.Use above method
Culture, respectively on March 25th, 2017, on April 22nd, 2017, the sunrise transplantation of seedlings in June 22 of on May 20th, 2017,2017, into
Motility rate statistics such as table 6 below：
Table 6：The transplanting survival rate of the tissue-cultured seedling of embodiment 1
As can be seen from Table 6, the young plant survival rate that each date transplants is more than 85%, and growth is quick；And May
Part, it is that transplanting survival rate is best on overburden soil with fertile soil 70%+ perlites 30%.
Further bletilla bulb tissue culture method described in embodiment 1 is contrasted with existing method, obtains as shown in table 7
Conclusion：
The comparison of the existing method of table 7 and the methods described of embodiment 1
In table " --- " indicate without
Can further it be reflected by table 7, this method simplifies the production link of bletilla pseudobulb tissue-cultured seedling, the false squama of induction
Stem, promotion pseudobulb fast-growth, and Rooting and hardening-off culture are completed in same step, are 90 days from seed to required time is transplanted
It is interior, incubation time is shortened, pseudobulb differentiation rate is improved, the fast-growth of pseudobulb is promoted, the shifting of tissue-cultured seedling is improved
Plant survival rate.
Finally illustrate, preferred embodiment above is merely illustrative of the technical solution of the present invention and unrestricted, although logical
Cross above preferred embodiment the present invention is described in detail, it is to be understood by those skilled in the art that can be
Various changes are made to it in form and in details, without departing from claims of the present invention limited range.
Claims (6)
1. a kind of special culture media of rapid induction bletilla bulb, it is characterised in that the special culture media is sprouted by bletilla seed
Culture medium and bulb efficiently induction and strong seedling culture base composition are sent out, wherein：
The formula of the bletilla seed germination medium is：1/2MS, NAA0.1mg/L, GA30.1mg/L, 6.5~7.0g/ of agar
L, white sugar 20g/L, pH5.8~6.0；
The bulb is efficiently induced and the formula of strong seedling culture base is：1/2MS, NAA0.5~0.6mg/L, 6-BA0.6~
0.8mg/L, PP3330.4~0.5mg/L, white sugar 30-45g/L, 25~40g/L of banana puree, activated carbon 0.1g/L, pH5.8~
6.0。
2. a kind of special culture media of rapid induction bletilla bulb according to claim 1, it is characterised in that the bulb is high
Effect is induced and the formula of strong seedling culture base is：1/2MS, NAA0.6mg/L, 6-BA0.8mg/L, PP3330.5mg/L, white sugar 40g/
L, banana puree 25g/L, activated carbon 0.1g/L, pH5.8-6.0.
3. a kind of tissue culture method of rapid induction bletilla bulb, it is characterised in that comprise the following steps：
1) sterilization of bletilla seed：Uncracked bletilla seed mass fraction is washed into surface 1- for 75% alcohol painting
Fruit pod, is then placed in 0.1% mercuric chloride 15~20min of immersion by 3min on superclean bench, then with aseptic water washing 3~
5 times, aseptic filter paper blots surface moisture in case inoculation；
2) under sterile condition of work, by step 1) seed that the bletilla seed of sterilization is inoculated into described in claim 1 sprouts
Send out in culture medium and cultivate；
3) when step 2) the bletilla seed sprouts the bulb being forwarded to during 0.8~1.2cm of height of seedling described in claim 1 and efficiently lures
Lead and strong seedling culture base in cultivate.
4. a kind of tissue culture method of rapid induction bletilla bulb according to claim 3, it is characterised in that step 2) culture
Condition is：Illumination 1500~2000Lx, 10~12h/ days；Temperature, 24~28 DEG C.
5. a kind of tissue culture method of rapid induction bletilla bulb according to claim 3, it is characterised in that step 3) culture
Condition is：Illumination 1500~2000Lx, 12~15h/ days；Temperature, 24~28 DEG C.
6. a kind of tissue culture method of rapid induction bletilla bulb according to claim 3, it is characterised in that the bletilla seed
Collection and storage practice be：Collection bletilla in 9~October is ripe, the place that full Fruit pod is placed in shady and cool ventilation, spreading for cooling 2~3 weeks；
When Fruit pod epidermis is changed into light brown, Fruit pod is loaded in kraft paper bag and is stored in 4 DEG C of refrigerating chambers of refrigerator, it is ensured that bletilla group
Train used in seedling whole year production.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108541593A (en) * | 2018-06-27 | 2018-09-18 | 芜湖东源新农村开发股份有限公司 | The method for tissue culture of white taro |
| CN109964815A (en) * | 2019-03-26 | 2019-07-05 | 成都大学 | A kind of bletilla striata aseptic seedling rapid induction Multiple Buds and fast numerous method of taking root |
| CN109964797A (en) * | 2019-03-22 | 2019-07-05 | 浙江大学 | The method for promoting bletilla pseudobulb to be proliferated/expand |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102246695A (en) * | 2011-05-10 | 2011-11-23 | 云南植物药业有限公司 | Method for preparing common bletilla pseudobulb in culture vessel and special culture media thereof |
-
2017
- 2017-07-07 CN CN201710552200.1A patent/CN107232061B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102246695A (en) * | 2011-05-10 | 2011-11-23 | 云南植物药业有限公司 | Method for preparing common bletilla pseudobulb in culture vessel and special culture media thereof |
Non-Patent Citations (3)
| Title |
|---|
| 张燕等: "不同培养基上白芨的种子萌发与幼苗形态发生", 《西北植物学报》 * |
| 王晓丽等: "活性炭和多效唑对垂花百合试管鳞茎膨大的影响", 《安徽农业科学》 * |
| 胡孔峰等: "《植物组织培养技术及应用》", 31 March 2006 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108541593A (en) * | 2018-06-27 | 2018-09-18 | 芜湖东源新农村开发股份有限公司 | The method for tissue culture of white taro |
| CN109964797A (en) * | 2019-03-22 | 2019-07-05 | 浙江大学 | The method for promoting bletilla pseudobulb to be proliferated/expand |
| CN109964815A (en) * | 2019-03-26 | 2019-07-05 | 成都大学 | A kind of bletilla striata aseptic seedling rapid induction Multiple Buds and fast numerous method of taking root |
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