CN106106167B - A kind of quick breeding method for tissue culture of oecoeclades falcata - Google Patents
A kind of quick breeding method for tissue culture of oecoeclades falcata Download PDFInfo
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- CN106106167B CN106106167B CN201610560836.6A CN201610560836A CN106106167B CN 106106167 B CN106106167 B CN 106106167B CN 201610560836 A CN201610560836 A CN 201610560836A CN 106106167 B CN106106167 B CN 106106167B
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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 invention discloses a kind of quick breeding method for tissue culture of oecoeclades falcata, choose explant of the blade as tissue culture of oecoeclades falcata；Bulbil Fiber differentiation is carried out to blade and forms protocorms；Protocorms are gradually differentiated to form adventitious bud by recycling generation and occurring repeatedly carry out Effective multiplication；Adventitious bud rooting culture carries out rooting culture after forming adventitious root, then normal culture.The quick breeding method for tissue culture of oecoeclades falcata of the present invention; solve the problems, such as that the breeding coefficient of oecoeclades falcata is low, growth rate is slow; the high-efficiency regeneration system of oecoeclades falcata is established, this method is not limited the industrial massive production, it can be achieved that high quality seedling by area, season, weather.
Description
Technical field
The invention belongs to technical field of plant propagation, specifically, being related to a kind of tissue-culturing quick-propagation side of oecoeclades falcata
Method.
Background technology
Oecoeclades falcata (Neofinetia falcate) is the perennial epiphytic orchid of orchid family oecoeclades falcata category (Neofinetia spp), strain
Type is small and exquisite, and blade leathery is verdant, and plump meat, flower pattern is peculiar, petal warp, like carrying out beautiful smart, pattern windward
Spotless white, the fragrance of a flower is charming, a burst of delicate fragrance, and flourishing aerial root is also striking；The root of oecoeclades falcata, stem, leaf and flower all have compared with
High ornamental value, and the polluter in environment can be absorbed, air is purified, taste is quiet and beautiful, smells comfortable pleasant, graceful bearing height
Refined uniqueness, also referred to as " pavilion is blue " " rich and honour blue ", also there is certain medical value.
Oecoeclades falcata has a large amount of cultivations in Japan, South Korea etc., and China's cultivation is less, but increasingly has been favored by people and look steadily
Mesh, market potential is huge, has a extensive future.If continuing to use traditional division propagation method, breeding coefficient is low, and growth rate is slow, much
The needs of people cannot be met；If being sowed with seed, there is ateliosis in seed again, it is difficult to sprout, variation variation is big, it is difficult to
Preserve the merit of parent；The breeding coefficient of oecoeclades falcata is low, becomes the bottleneck for restricting its research and development.
Invention content
In view of this, the present invention for oecoeclades falcata breeding coefficient is low, problem that growth rate is slow, provide a kind of oecoeclades falcata
Quick breeding method for tissue culture establishes the high-efficiency regeneration system of oecoeclades falcata, it can be achieved that the industrial massive of high quality seedling produces.
In order to solve the above-mentioned technical problem, the invention discloses a kind of quick breeding method for tissue culture of oecoeclades falcata, choose
Explant of the blade of oecoeclades falcata as tissue culture；Bulbil Fiber differentiation is carried out to blade and forms protocorms；To protocorms into
Row Multiplying culture, protocorms are gradually differentiated to form adventitious bud by recycling generation and occurring repeatedly carry out Effective multiplication；No
Normal bud and protocorms culture of rootage carry out rooting culture after forming adventitious root, then normal culture.
Further, following steps are specifically included：
Step 1, the selection and processing of explant
The blade for choosing oecoeclades falcata is starting material, as the explant of tissue culture after disinfection；
Step 2, the induction of protocorms
It is cultivated being inoculated on protocorms inducing culture MS+TDZ5.0mg/L by the explant of disinfection, leaf
Piece base portion gradually forms embryo callus, and embryo callus gradually forms protocorms；
Step 3, the proliferation of protocorms
The protocorms that step 2 is formed are transferred on protocorms proliferated culture medium MS+6BA2.0mg/L and are cultivated,
Protocorms occur to realize Effective multiplication by occurring repeatedly and recycling, and gradually form adventitious bud；
Step 4, seedling of taking root and rooting culture
The adventitious bud induced and protocorms are transferred on root media MS+IBA1.0mg/L and cultivated, 20
After~30 days, a plurality of meat adventitious root is grown, seedling of taking root becomes new plant, normally cultivated after hardening, slow seedling.
Further, the condition of culture of protocorms Fiber differentiation, protocorms Multiplying culture and culture of rootage is：Training
25 DEG C of temperature is supported, light application time is 12~14h/d, and intensity of illumination is 40~60u.Mol.m-2.s-1。
Further, protocorms Fiber differentiation, protocorms Multiplying culture and culture of rootage, minimal medium used
It is MS culture mediums, dosage of sucrose is 20~40g/L, and coagulator is agar powder, dosage is 6~7g/L, activated carbon 2g/L, coconut
Juice additive amount is 10%, medium pH 5.8.
Further, the hardening, slow seedling are specially：First in tissue culture chamber opening bottle cap hardening 3 days, it is then transported to temperature
Room or cool canopy, finishing scouring seedling 3 days；Then tissue-cultured seedling is taken out from culture bottle, cleans the agar on its surface, is transplanted to culture substrate
Middle carry out slow seedling, then normal culture.
Further, the culture substrate is moss.
Further, within the culture substrate water content 25%, relative air humidity 80~90%, shading 50%.
Compared with prior art, the present invention can be obtained including following technique effect：
(1) present invention is used as the explant of tissue culture by choosing the blade of oecoeclades falcata；Bulbil Fiber differentiation is carried out to blade
Form protocorms；Protocorms are gradually differentiated to form adventitious bud by recycling generation and occurring repeatedly carry out Effective multiplication；
Adventitious bud and protocorms culture of rootage carry out rooting culture after forming adventitious root, and then normal culture, solves the numerous of oecoeclades falcata
The problem that coefficient is low, growth rate is slow is grown, the high-efficiency regeneration system of oecoeclades falcata is established；
(2) numerous soon by leaf tissue culture progress oecoeclades falcata, it is not limited, it can be achieved that high quality seedling by area, season, weather
Industrial massive production, and inhereditary feature is with respect to stable and consistent；
(3) it is the genetic conversion system for improving oecoeclades falcata, establishes the platform of molecular breeding, heredity is carried out to it from molecular level
Improvement, further investigation are developed and used and are laid a good foundation.
Certainly, it implements any of the products of the present invention it is not absolutely required to while reaching all the above technique effect.
Description of the drawings
Attached drawing described herein is used to provide further understanding of the present invention, and constitutes the part of the present invention, this hair
Bright illustrative embodiments and their description are not constituted improper limitations of the present invention for explaining the present invention.In the accompanying drawings：
Fig. 1 is the tissue-culturing quick-propagation process comparison diagram of oecoeclades falcata of the embodiment of the present invention；In figure, A is oecoeclades falcata blade base
Portion's induced synthesis embryo callus and protocorms, B are the proliferation of protocorms, and C is that protocorms grow blade and indefinite
Bud, D are that protocorms grow fleshy root, and E is that oecoeclades falcata is taken root seedling, and F is oecoeclades falcata potting.
Specific implementation mode
Carry out the embodiment that the present invention will be described in detail below in conjunction with embodiment, thereby 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.
A kind of quick breeding method for tissue culture of oecoeclades falcata of the present invention, specifically includes following steps：
Step 1, the selection and processing of explant
The blade for choosing oecoeclades falcata is starting material, as the explant of tissue culture after disinfection；
Specifically, described sterilize is specially：Oecoeclades falcata blade is cleaned into 30min with liquid detergent solution, is during which constantly gently stirred
It mixes, then flowing water rinses 30~60min, is transferred to the above-mentioned oecoeclades falcata blade cleaned on sterilized superclean bench
In sterile beaker, the alcohol for being 75% with volumetric concentration impregnates 30~60s, during which constantly shakes beaker, pours out alcohol, then use matter
The mercuric chloride for measuring concentration 0.1% impregnates 3~5min, constantly gently shakes during immersion, after pouring out mercuric chloride, is passed through with sterile water wash
The blade 3 times or more of disinfection.
Step 2, the induction of protocorms
On superclean bench, the explant Jing Guo surface sterilization is transferred in aseptic inoculation disk, is inhaled with aseptic filter paper
Blade base is downwardly into protocorms inducing culture MS+ by dry explant surface moisture with sterilized scissors
TDZ5.0mg/L(TDZ：Thidiazuron) on cultivated, 20d rear blade base portions formed yellow embryo callus, after 20d
Afterwards, embryo callus is green by xanthochromia, forms small green ball, protocorms are formed, as shown in Fig. 1 (A)；
Condition of culture：25 DEG C of cultivation temperature, light application time are 12~14h/d, and intensity of illumination is 40~60u.Mol.m-2.s-1。
Protocorms (Protocorm-like body, PLB) are the intergrants of embryo's generation and allelotaxis, are a kind of
Complex tissue is very effective regeneration modes of reproduction, and protocorms can be proliferated, and occur repeatedly and recycle hair
Raw, protocorms have apparent " bipolarity ", " can sprout above, take root below " simultaneously, form complete plant.
Step 3, the proliferation of protocorms
The protocorms that step 2 is formed are transferred to protocorms proliferated culture medium MS+6BA2.0mg/L (6BA：Benzyl amino
Purine) on cultivated, protocorms by occur repeatedly and recycle occur realize Effective multiplication, as shown in Fig. 1 (B), and by
Blade and adventitious bud are gradually grown, as shown in Fig. 1 (C)；
Condition of culture：25 DEG C of cultivation temperature, light application time are 12~14h/d, and intensity of illumination is 40~60u.Mol.m-2.s-1。
Step 4, seedling of taking root and rooting culture
The adventitious bud induced and protocorms are transferred to root media MS+IBA1.0mg/L (IBA：Indolebutyric acid)
On cultivated, after 20~30 days, grow a plurality of meat adventitious root, as shown in Fig. 1 (D), seedling of taking root become new plant, such as
Shown in Fig. 1 (E), normally cultivated after hardening, slow seedling, as shown in Fig. 1 (F).
Condition of culture is：25 DEG C of cultivation temperature, light application time be 12~14h/d, intensity of illumination be 40~
60u.Mol.m-2.s-1。
Wherein, protocorms Fiber differentiation, protocorms Multiplying culture and culture of rootage, minimal medium used are
MS culture mediums, dosage of sucrose are 20~40g/L, and coagulator is agar powder, dosage is 6~7g/L, activated carbon 2g/L, and coconut milk adds
Dosage is 10%, medium pH 5.8.
Further, the hardening, slow seedling are specially：First in tissue culture chamber opening bottle cap hardening 3 days, it is then transported to temperature
Room or cool canopy, finishing scouring seedling 3 days；Then tissue-cultured seedling is taken out from culture bottle, cleans the agar on its surface, is transplanted to culture substrate
Middle carry out slow seedling, then normal culture.
Preferably, culture substrate is moss, and within culture substrate water content 25%, relative air humidity 80~90% hides
Light 50%.
The present invention generates protocorms using oecoeclades falcata blade base under the basic element of cell division of high concentration, and protocorms both may be used
The characteristics of to occur repeatedly and recycle generation, and become well established and vigorously developing high quality seedling because it can induce with bipolarity, with class
Protocorm is core, and protocorms become the target of regulation and control, is realized by regulating and controlling growth, differentiation and the development of protocorms excellent
The large-scale production of matter seedling, and inhereditary feature is with respect to stable and consistent.
Embodiment 1
Step 1, the selection and processing of explant
The blade for choosing oecoeclades falcata is starting material, as the explant of tissue culture after disinfection；
Step 2, the induction of protocorms
On superclean bench, the explant Jing Guo surface sterilization is transferred in aseptic inoculation disk, is inhaled with aseptic filter paper
Blade base is downwardly into protocorms inducing culture MS+ by dry explant surface moisture with sterilized scissors
TDZ5.0mg/L(TDZ：Thidiazuron) on cultivated, 20d rear blade base portions formed yellow embryo callus, after 20d
Afterwards, green by xanthochromia, small green ball is formed, protocorms are formed；
Step 3, the proliferation of protocorms
The protocorms that step 2 is formed are transferred to protocorms proliferated culture medium MS+6BA2.0mg/L (6BA：Benzyl amino
Purine) on cultivated, protocorms by occur repeatedly and recycle occur realize proliferation, gradually grow blade and adventitious bud；
Step 4, seedling of taking root and rooting culture
The adventitious bud induced and protocorms are transferred to root media MS+IBA1.0mg/L (IBA：Indolebutyric acid)
On cultivated, gradually grow a plurality of meat adventitious root, seedling of taking root becomes new plant, by plant first in tissue culture chamber opening bottle
Lid hardening 3 days is then transported to greenhouse or cool canopy, finishing scouring seedling 3 days；Then tissue-cultured seedling is taken out from culture bottle, cleans it
The agar on surface is transplanted in culture substrate (moss), culture substrate water content 25%, relative air humidity 80%, shading
50%, slow seedling is carried out, then normal culture.
Wherein, the condition of culture of protocorms Fiber differentiation, protocorms Multiplying culture and culture of rootage is：Culture temperature
25 DEG C, light application time 12h/d, intensity of illumination 60u.Mol.m of degree-2.s-1。
Wherein, protocorms Fiber differentiation, protocorms Multiplying culture and culture of rootage, minimal medium used are
MS culture mediums, dosage of sucrose 20g/L, coagulator are agar powder, dosage 6g/L, activated carbon 2g/L, and coconut milk additive amount is
10%, medium pH 5.8.
Embodiment 2
Step 1, the selection and processing of explant
The blade for choosing oecoeclades falcata is starting material, as the explant of tissue culture after disinfection；
Step 2, the induction of protocorms
On superclean bench, the explant Jing Guo surface sterilization is transferred in aseptic inoculation disk, is inhaled with aseptic filter paper
Blade base is downwardly into protocorms inducing culture MS+ by dry explant surface moisture with sterilized scissors
TDZ5.0mg/L(TDZ：Thidiazuron) on cultivated, 20d rear blade base portions formed yellow embryo callus, after 20d
Afterwards, green by xanthochromia, small green ball is formed, protocorms are formed；
Step 3, the proliferation of protocorms
The protocorms that step 2 is formed are transferred to protocorms proliferated culture medium MS+6BA2.0mg/L (6BA：Benzyl amino
Purine) on cultivated, protocorms by occur repeatedly and recycle occur realize proliferation, gradually grow blade and adventitious bud；
Step 4, seedling of taking root and rooting culture
The adventitious bud induced and protocorms are transferred to root media MS+IBA1.0mg/L (IBA：Indolebutyric acid)
On cultivated, after 20~30 days, grow a plurality of meat adventitious root, seedling of taking root becomes new plant, by plant first in tissue culture
Chamber opening bottle cap hardening 3 days is then transported to greenhouse or cool canopy, finishing scouring seedling 3 days；Then tissue-cultured seedling is taken from culture bottle
Go out, cleans the agar on its surface, be transplanted in culture substrate (moss), culture substrate water content 23%, relative air humidity
85%, shading 50% carries out slow seedling, then normal culture.
Wherein, the condition of culture of protocorms Fiber differentiation, protocorms Multiplying culture and culture of rootage is：Culture temperature
25 DEG C, light application time 14h/d, intensity of illumination 40u.Mol.m of degree-2.s-1。
Wherein, protocorms Fiber differentiation, protocorms Multiplying culture and culture of rootage, minimal medium used are
MS culture mediums, dosage of sucrose 30g/L, coagulator are agar powder, dosage 6.5g/L, activated carbon 2g/L, coconut milk additive amount
It is 10%, medium pH 5.8.
Embodiment 3
Step 1, the selection and processing of explant
The blade for choosing oecoeclades falcata is starting material, as the explant of tissue culture after disinfection；
Step 2, the induction of protocorms
On superclean bench, the explant Jing Guo surface sterilization is transferred in aseptic inoculation disk, is inhaled with aseptic filter paper
Blade base is downwardly into protocorms inducing culture MS+ by dry explant surface moisture with sterilized scissors
TDZ5.0mg/L(TDZ：Thidiazuron) on cultivated, 20d rear blade base portions formed yellow embryo callus, after 20d
Afterwards, green by xanthochromia, small green ball is formed, protocorms are formed；
Step 3, the proliferation of protocorms
The protocorms that step 2 is formed are transferred to protocorms proliferated culture medium MS+6BA2.0mg/L (6BA：Benzyl amino
Purine) on cultivated, protocorms by occur repeatedly and recycle occur realize proliferation, gradually grow blade and adventitious bud；
Step 4, seedling of taking root and rooting culture
The adventitious bud induced and protocorms are transferred to root media MS+IBA1.0mg/L (IBA：Indolebutyric acid)
On cultivated, after 20~30 days, grow a plurality of meat adventitious root, seedling of taking root becomes new plant, first in tissue culture chamber opening
Bottle cap hardening 3 days is then transported to greenhouse or cool canopy, finishing scouring seedling 3 days；Then tissue-cultured seedling is taken out from culture bottle, is cleaned
The agar on its surface is transplanted in culture substrate (moss), culture substrate water content 20%, relative air humidity 90%, shading
50%, slow seedling is carried out, then normal culture.
Wherein, the condition of culture of protocorms Fiber differentiation, protocorms Multiplying culture and culture of rootage is：Culture temperature
25 DEG C, light application time 13h/d, intensity of illumination 50u.Mol.m of degree-2.s-1。
Wherein, protocorms Fiber differentiation, protocorms Multiplying culture and culture of rootage, minimal medium used are
MS culture mediums, dosage of sucrose 40g/L, coagulator are agar powder, dosage 7g/L, activated carbon 2g/L, and coconut milk additive amount is
10%, medium pH 5.8.
Embodiment 4
Step 1, the selection and processing of explant
The blade for choosing oecoeclades falcata is starting material, as the explant of tissue culture after disinfection；
Step 2, the induction of protocorms
On superclean bench, the explant Jing Guo surface sterilization is transferred in aseptic inoculation disk, is inhaled with aseptic filter paper
Blade base is downwardly into protocorms inducing culture MS+ by dry explant surface moisture with sterilized scissors
TDZ5.0mg/L(TDZ：Thidiazuron) on cultivated, 20d rear blade base portions formed yellow embryo callus, after 20d
Afterwards, green by xanthochromia, small green ball is formed, protocorms are formed；
Step 3, the proliferation of protocorms
The protocorms that step 2 is formed are transferred to protocorms proliferated culture medium MS+6BA2.0mg/L (6BA：Benzyl amino
Purine) on cultivated, protocorms by occur repeatedly and recycle occur realize proliferation, gradually grow blade and adventitious bud；
Step 4, seedling of taking root and rooting culture
The adventitious bud induced and protocorms are transferred to root media MS+IBA1.0mg/L (IBA：Indolebutyric acid)
On cultivated, after 20~30 days, grow a plurality of meat adventitious root, seedling of taking root becomes new plant, first in tissue culture chamber opening
Bottle cap hardening 3 days is then transported to greenhouse or cool canopy, finishing scouring seedling 3 days；Then tissue-cultured seedling is taken out from culture bottle, is cleaned
The agar on its surface is transplanted in culture substrate (moss), culture substrate water content 18%, relative air humidity 85%, shading
50%, slow seedling is carried out, then normal culture.
Wherein, the condition of culture of protocorms Fiber differentiation, protocorms Multiplying culture and culture of rootage is：Culture temperature
25 DEG C, light application time 13h/d, intensity of illumination 55u.Mol.m of degree-2.s-1。
Wherein, protocorms Fiber differentiation, protocorms Multiplying culture and culture of rootage, minimal medium used are
MS culture mediums, dosage of sucrose 25g/L, coagulator are agar powder, dosage 6.8g/L, activated carbon 2g/L, coconut milk additive amount
It is 10%, medium pH 5.8.
Oecoeclades falcata leaf tissue culture is bred using the propagation method of protocorms of the present invention and uses traditional plant division numerous
The data comparison for growing the breeding coefficient that method breeds oecoeclades falcata is shown in Table 1：
1 propagation method of the present invention of table and influence of the Traditional breeding processes to oecoeclades falcata growth coefficient
As can be seen from Table 1, propagation method using the present invention breeds oecoeclades falcata, and geometry base is presented in growth coefficient
Numerical expression increases, significantly larger than traditional division propagation mode, numerous soon by leaf tissue culture progress oecoeclades falcata using the above method,
Breeding coefficient is high, and growth rate is fast, and the industrial massive life, it can be achieved that high quality seedling is not limited by area, season, weather
Production.
Some vocabulary has such as been used to censure special component or method in specification and claim.Art technology
Personnel are, it is to be appreciated that different regions may call the same ingredient with different nouns.This specification and claims are not
In such a way that the difference of title is used as and distinguishes ingredient.As the "comprising" of the specification in the whole text and claim mentioned in is
One open language, therefore should be construed to " including but not limited to "." substantially " refer to this field in receivable error range
Technical staff can solve the technical problem within a certain error range, basically reach the technique effect.Specification is follow-up
It is described as implementing the better embodiment of the present invention, so description is for the purpose of the rule for illustrating the present invention, not
To limit the scope of the present invention.Protection scope of the present invention is when subject to appended claims institute defender.
It should also be noted that, the terms "include", "comprise" or its any other variant are intended to nonexcludability
Including so that commodity or system including a series of elements include not only those elements, but also include not clear
The other element listed, or further include for this commodity or the intrinsic element of system.In the feelings not limited more
Under condition, the element that is limited by sentence "including a ...", it is not excluded that including the element commodity or system in also
There are other identical elements.
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, is not to be taken as excluding other embodiments, and can be used for various other combinations, modification
And environment, and can be carried out by the above teachings or related fields of technology or knowledge in the scope of the invention is set forth herein
Change.And changes and modifications made by those skilled in the art do not depart from the spirit and scope of invention, then should all be weighed appended by invention
In the protection domain that profit requires.
Claims (4)
1. a kind of quick breeding method for tissue culture of oecoeclades falcata, which is characterized in that choose the blade of oecoeclades falcata as the outer of tissue culture
Implant；Fiber differentiation is carried out to blade and forms protocorms；Protocorms occur and occur repeatedly effectively to be increased by recycling
It grows, and is gradually differentiated to form adventitious bud；Adventitious bud and protocorms culture of rootage carry out rooting culture after forming adventitious root, then
Normal culture；Specifically include following steps：
Step 1, the selection and processing of explant
The blade for choosing oecoeclades falcata is starting material, as the explant of tissue culture after disinfection；
Step 2, the induction of protocorms
It is cultivated being inoculated on protocorms inducing culture MS+TDZ5.0mg/L by the explant of disinfection, blade base
Portion gradually forms embryo callus, and embryo callus gradually forms protocorms；
Step 3, the proliferation of protocorms
The protocorms that step 2 is formed are transferred on protocorms proliferated culture medium MS+6-BA2.0mg/L and are cultivated, class
Protocorm occurs to realize Effective multiplication by occurring repeatedly and recycling, and gradually forms adventitious bud；
Step 4, seedling of taking root and rooting culture
The adventitious bud induced and protocorms are transferred on root media MS+IBA1.0mg/L and cultivated, 20~30
After it, a plurality of meat adventitious root is grown, seedling of taking root becomes new plant, normally cultivated after hardening, slow seedling；
Wherein, the condition of culture of protocorms Fiber differentiation, protocorms Multiplying culture and culture of rootage is：Cultivation temperature 25
DEG C, light application time is 12~14h/d, and intensity of illumination is 40~60 μm of olm-2·s-1；
Wherein, protocorms Fiber differentiation, protocorms Multiplying culture and culture of rootage, minimal medium used are MS trainings
Base is supported, dosage of sucrose is 20~40g/L, and coagulator is agar powder, dosage is 6~7g/L, activated carbon 2g/L, coconut milk additive amount
It is 10%, medium pH 5.8.
2. the quick breeding method for tissue culture of oecoeclades falcata as described in claim 1, which is characterized in that the hardening, slow seedling tool
Body is：First in tissue culture chamber opening bottle cap hardening 3 days, it is then transported to greenhouse or cool canopy, finishing scouring seedling 3 days；Then by tissue-cultured seedling
It is taken out from culture bottle, cleans the agar on its surface, be transplanted in culture substrate and carry out slow seedling, then normal culture.
3. the quick breeding method for tissue culture of oecoeclades falcata as claimed in claim 2, which is characterized in that the culture substrate is tongue
Moss.
4. the quick breeding method for tissue culture of oecoeclades falcata as claimed in claim 2 or claim 3, which is characterized in that the culture substrate
Within water content 25%, relative air humidity 80~90%, shading 50%.
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