CN110100737A - A kind of candidum tissue culturing rapid propagation method - Google Patents
A kind of candidum tissue culturing rapid propagation method Download PDFInfo
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- CN110100737A CN110100737A CN201910542585.2A CN201910542585A CN110100737A CN 110100737 A CN110100737 A CN 110100737A CN 201910542585 A CN201910542585 A CN 201910542585A CN 110100737 A CN110100737 A CN 110100737A
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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 candidum tissue culturing rapid propagation methods, comprising the following steps: the preparation of capsule seed, the induction of seed are sprouted and differentiation, the proliferation of seedling, strengthening seedling and rooting and transplanting.The present invention is mainly that three step strong sprout methods of the Fiber differentiation of seed, seedling proliferation culture, strengthening seedling and rooting culture is used to substitute candidum tissue culturing traditional four-step method of proliferating, seedling can effectively shorten two to three months production cycles, to reduce the time of each link, manpower, feed consumption cost in production；Within each time for expanding about the last fortnight in numerous link, the method for pure dark culture is all made of to cultivate, can effectively promote the speed of growth, using three generations's method of proliferating, so that the gene in maternal plant is more preferably replicated, ensure that the pure of seedling kind；The formula for improving culture medium, by increasing potash fertilizer and siliceous fertilizer, so that sturdy degree, the uniformity of bottle seedling growth are improved especially in the strong sprout stage, enhancing kind shoot survival percent promotes the market competitiveness of seedling.
Description
Technical field
The present invention relates to technical field of Chinese herbal medicine cultivation, specifically a kind of candidum tissue culturing rapid propagation method.
Background technique
Dendrobium candidum is a kind of traditional rare rare traditional Chinese medicine, because having antitumor, anti-aging, acoustic resistive band fatigue, expanding
The effects of opening blood vessel and enhancing human immunity, referred to as " help mesona ", rank first of " Chinese nine big mesonas ".With iron sheet
Dendrobium nobile health value is continuously developed, and medicinal material market is also gradually increasing the demand of dendrobium candidum, but due to excessively adopting for a long time
It plucks, Chinese wild resource is exhausted, and low with traditional vegetative manner breeding coefficient, is unable to satisfy commerial growing
Seedling needs, and tissue-cultured seedling has become the main source of seedling of current dendrobium candidum plantation at present.Currently to candidum tissue culturing
Fast breeding technique research is more, but most of there are medium components complicated, growing-seedling period compared with long, labor intensive is more, tissue-cultured seedling at
The problems such as this is high seriously constrains the industrialized development of dendrobium candidum.To sum up, it needs to improve the prior art.
Summary of the invention
It is fast the technical problem to be solved by the present invention is in view of the above shortcomings of the prior art, provide a kind of candidum tissue culturing
Fast propagation method, compared with prior art can shorter growing-seedling period and cost acquisition candidum tissue culturing using method of the invention
Seedling.
To achieve the above object, the present invention provides candidum tissue culturing rapid propagation method technical solution are as follows: a kind of iron sheet
Dendrobe tissue culture rapid propagation method, successively the following steps are included:
1) it, draws materials: selecting the uncracked mature capsule tied on the dendrobium candidum of robust growth, sterile washing lotion is used to wash
Only, drain rear spare；
2), seed induction sprout and differentiation: by the resulting capsule of step 1) it is sterilized after, cut off each 0 .3 in its head and the tail both ends~
It is longitudinally splitted after 0 .7cm, takes out seed, seed is uniformly inoculated into seed and is sprouted on differential medium, is carried out in tissue culture room
Culture；
Condition of culture is set are as follows: induction initial stage 13~15 days carries out dark culture, and temperature setting is 24~26 DEG C, humidity 60~
70%；Later period 43~47 days, illumination and dark culture alternate culture are carried out, temperature setting is 24~26 DEG C, humidity 60~70%, light
It is 8h/d, intensity of illumination 1000-1200lux, 16 hours dark cultures according to the time；Culture terminates to obtain protocorm；
3), the proliferation of seedling: step 2) protocorm obtained will be chosen that growing way is vigorous, uniform protocorm, carried out primary
Expand numerous, coefficient is set as 95~105 times, and uniformly switching carries out Multiplying culture in seedling proliferation culture medium；
Condition of culture is set are as follows: proliferation initial stage 13~15 days carries out dark culture, and temperature setting is 24~26 DEG C, humidity 60~
70%；Later period 45~75 days, illumination and dark culture alternate culture are carried out, temperature setting is 24~26 DEG C, humidity 60~70%, light
It is 10h/d, intensity of illumination 2000-2500lux, 14 hours dark cultures according to the time；Culture terminates to obtain tufted seedling；
4), strengthening seedling and rooting: the tufted seedling that step 3) Multiplying culture is obtained chooses vigorous, uniform, the long extremely proliferation bottle of growing way and binds
Tufted seedling is split from base portion and is expanded for the second time numerous by high (i.e. 2~3cm), and coefficient is set as 7-~9 times, uniformly transfer
Strengthening seedling and rooting culture is carried out on to strengthening seedling and rooting culture medium；
Condition of culture is set are as follows: strong sprout initial stage 13~15 days, dark culture is carried out, temperature setting is 24~26 DEG C, humidity 60~
70%；Later period 65~75 days, illumination and dark culture alternate culture are carried out, temperature setting is 24~26 DEG C, light application time 12h/
D, intensity of illumination 2800-3000lux, 12 hours dark cultures；Culture terminates to obtain strengthening seedling and rooting；
5), transplanting: the seedling length to strengthening seedling and rooting culture acquisition to 4~5cm high and grows root of the length more than or equal to 2cm extremely
When few 3 roots, acquisition can bottle outlet plant seedling, carry out the outdoor hardening of two weeks by a definite date, can bottle outlet plant seedling from tissue culture
It is taken out in bottle, root is removed using clear water and carries culture medium, can be obtained plantation seedling.
Candidum tissue culturing enrichment procedure in compared to the prior art, the present invention are arranged the proliferation of seedling by step 2
The three step strong sprout methods to four realize, and about two weeks were all made of pure dark training before being arranged in that dendrobium candidum is each and expanding numerous link
Feeding method is cultivated, and dark culture method therein is that inventor is obtained by long-term multiple groups bottle seedling culture correlation data, and inventor is led to
Cross it was found that expanding numerous link early period, the bottle seedling of dark culture has no effect on seedling early growth, by dark culture early period and later period illumination,
The combination culture of dark culture alternate culture, the speed of growth compared to the prior art in pure illumination, the comparison of dark culture alternate culture
Bottle seedling faster, and bottle seedling to grow into the comparison bottle seedling of neat rate and leaf color also than illumination, dark culture alternate culture more excellent,
In the early stage under dark culture state, seedling can freely be grown up inventor's discovery according to habit, in conjunction with later period illumination, secretly
The growth that alternate culture increases bottle seedling is cultivated, integral into together can more preferably, other compared to the prior art illumination, dark culture alternating
Culture, the light application time in the illumination in each step later period of the invention, dark culture alternate culture is obvious less, thus contracting
The electric quantity consumption of electricity consumption illumination is greatly reduced under the premise of the short growth cycle time again, reduces production cost.
The further embodiment of quick breeding by group culture method as dendrobium candidum of the present invention: the seed in the step 2)
Sprout differential medium are as follows: MS+NAA0.25~0.35mg/L+ sucrose 25~35g/L+, 5~10g/L+ of agar potato 15~
20g/L, pH 6.3.
The further embodiment of quick breeding by group culture method as dendrobium candidum of the present invention: the seedling in the step 3)
Proliferated culture medium are as follows: MS+NAA0.45~0.5mg/L+ sucrose 25~35g/L+ agar 5~10g/L, pH are 6.4~6.5.
The further embodiment of quick breeding by group culture method as dendrobium candidum of the present invention: the strong sprout in the step 4)
Root media are as follows: MS+NAA0.35mg/L+ sucrose 25~35g/L+ agar 5~10g/L+ banana 110~120g/L, pH are
6.1~6.3.
The further embodiment of quick breeding by group culture method as dendrobium candidum of the present invention: also add in the MS culture medium
Add 0.35~0.45g/L of potash fertilizer, 0.15~0.25g/L of siliceous fertilizer.
Compared with prior art, the beneficial effects of the present invention are: the present invention is mainly to use Fiber differentiation, the seedling of seed
Multiplying culture, strengthening seedling and rooting Multiplying culture three step strong sprout methods come realize dendrobium candidum tissue culture proliferation, compared with the prior art
In four generations so that five generations be proliferated production method, have the advantages that
1), seedling can effectively shorten two to three months production cycles, thus reduce in production the time of each link, manpower,
Feed consumption cost；
2), within each time for expanding about the last fortnight in numerous link, the method for pure dark culture is all made of to cultivate, can effectively be mentioned
The speed of growth is risen, and bottle seedling grows into neat rate and leaf color more preferably, significantly reduces use under the premise of guaranteeing excellent
Electricity further reduces production cost；
3), using three generations's method of proliferating, according to Heredity theory, so that the gene in maternal plant is more preferably replicated, reduce mostly generation
Mutation probability, ensure that the pure of seedling kind, while also increase the survival rate of seedling；
4) formula for, improving culture medium, by increasing potash fertilizer and siliceous fertilizer, so that especially in the strong sprout stage, bottle seedling is grown thick
Strong degree, uniformity are improved, and promote the market competitiveness of seedling.
Specific embodiment
Below in conjunction with specific method in the embodiment of the present invention, technical solution in the embodiment of the present invention carry out it is clear,
It is fully described by, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Base
Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts it is all its
His embodiment, shall fall within the protection scope of the present invention.
The uncracked mature capsule that selection is tied on the dendrobium candidum with a collection of robust growth is arranged to four groups and waits cultivating kind
Son carries out tissue-cultured seedling culture, wherein using embodiment two, comparative example two of the method for the present invention.
Embodiment 1:
A kind of candidum tissue culturing rapid propagation method, includes the following steps:
1) it, draws materials: selecting the uncracked mature capsule in 120 days tied on the dendrobium candidum of this batch of robust growth, used nothing
Bacterium washing lotion cleans, it is rear spare to drain；
2), seed induction sprout and differentiation: by the resulting capsule of step 1) it is sterilized after, cut off each 0 .3 in its head and the tail both ends~
It is longitudinally splitted after 0 .7cm, takes out seed, 1 seed of embodiment that seed is uniformly inoculated into setting is sprouted on differential medium,
It is cultivated in tissue culture room；
Example 1 group trains the condition of culture of room setting are as follows: at 14 days of induction initial stage, dark culture is carried out, temperature setting is 25 DEG C,
Humidity 60%；Later time carries out illumination and dark culture alternate culture, and temperature setting is 25 DEG C, humidity 60%, and light application time is
8h/d, intensity of illumination 1000lux, 16 hours dark cultures；Protocorm is being obtained by culture in 60 days in total；
Sprouting differential medium therein is prepared by MS+NAA0.3mg/L+ sucrose 30g/L+ fine jade 5g/L+ potato 15g/L, pH setting
It is 6.3.
3), the proliferation of seedling: step 2) protocorm obtained will be chosen that growing way is vigorous, uniform protocorm, carried out
Once expand numerous, coefficient is set as 100 times, shifts spoon for protocorm seedling bale-out by dedicated protocorm, and transfer in seedling
It is proliferated in bottle, uniformly spills and carry out Multiplying culture in seedling proliferation culture medium in bottle；
It is provided with condition of culture are as follows: be proliferated 14 days of initial stage, carry out dark culture, temperature setting is 25 DEG C, humidity 60%；Later period
Illumination and dark culture alternate culture are carried out, temperature setting is 25 DEG C, humidity 60%, light application time 10h/d, and intensity of illumination is
2000lux, 14 hours dark cultures；The culture in 45 to 65 days of later period time-consuming terminates to obtain tufted seedling；
Seedling proliferation culture medium therein is pressed: the formula of MS+NAA0.5mg/L+ sucrose 35g/L+ agar 5g/L is prepared, pH setting
It is 6.5.
4), strengthening seedling and rooting: the tufted seedling that step 3) Multiplying culture is obtained, choose growing way it is vigorous, uniform, long to 2~
Tufted seedling is split from base portion and is expanded for the second time numerous by 3cm, and coefficient is set as 8 times, is uniformly transferred to equipped with strengthening seedling and rooting
It is upper in the strengthening seedling and rooting proliferation bottle of culture medium to carry out strengthening seedling and rooting culture；
The condition of culture being provided with are as follows: 14 days of strong sprout initial stage carry out dark culture, and temperature setting is 25 DEG C, humidity 60%；Afterwards
Time phase carries out illumination and dark culture alternate culture, and temperature setting is 25 DEG C, light application time 12h/d, and intensity of illumination is
2800lux, 12 hours dark cultures；The culture of time-consuming 65 days of later period obtains strengthening seedling and rooting；
Strengthening seedling and rooting culture medium therein is pressed: MS+NAA0.35mg/L+ sucrose 30g/L+ agar 10g/L+ banana 120g/L's matches
Side is prepared, and pH is set as 6.3
5), transplant: the seedling obtained according to step 3) strengthening seedling and rooting culture is grown to 4~5cm high, major part substantially grows length
Root three or more more than or equal to 2cm, acquisition can bottle outlet plant seedling, can bottle outlet plantation seedling taken out from tissue culture bottle, adopt
Root is removed with clear water and carries culture medium, can be obtained plantation seedling.
Wherein the basic used time of embodiment 1 reaches 93% in the plantation seedling planting percent that 198 day time completed to obtain, residue kind
Seedling also complies with requirement after carrying out a few days culture again, and planting percent is 99% in total, and whole seedling leaf color is bright and new beautiful, erects
Straight elevator pulls out, and growth curvature phenomenon is unobvious.
Embodiment 2:
A kind of candidum tissue culturing rapid propagation method, includes the following steps:
1) it, draws materials: selecting the uncracked mature capsule in 120 days tied on the dendrobium candidum of robust growth, used sterile wash
Liquid cleans, it is rear spare to drain；
2), seed induction sprout and differentiation: by the resulting capsule of step 1) it is sterilized after, cut off each 0 .3 in its head and the tail both ends~
It is longitudinally splitted after 0 .7cm, takes out seed, the seed that seed is uniformly inoculated into embodiment 2 is sprouted on differential medium, in group
It is cultivated training interior；
The wherein condition of culture of 2 tissue culture room of embodiment setting are as follows: 15 day time is arranged in induction initial stage, carries out dark culture, and temperature is set
24 DEG C are set to, humidity 70%；Stage carries out illumination and dark culture alternate culture, and temperature setting is 25 DEG C, humidity 70%, illumination
Time is 8h/d, intensity of illumination 1200lux, 16 hours dark cultures；Culture in time-consuming 44 days terminates to obtain protocorm；
Sprouting differential medium therein is prepared by MS+NAA0.25mg/L+ sugarcane 25g/L+ agar 10g/L+ potato 20g/L, and pH is set
It is set to 6.3.
3), the proliferation of seedling: step 2) protocorm obtained will be chosen that growing way is vigorous, uniform protocorm, carried out
Once expand numerous, coefficient is set as 95 times, and uniformly switching carries out Multiplying culture in seedling proliferation culture medium；
The condition of culture being provided with are as follows: proliferation initial stage is set as 15 days, carries out dark culture, and temperature setting is 25 DEG C, humidity
70%；Later time is set as 53 days, carries out illumination and dark culture alternate culture, and temperature setting is 25 DEG C, humidity 70%, illumination
Time is 10h/d, intensity of illumination 2500lux, 14 hours dark cultures；Culture terminates to obtain tufted seedling；
Seedling proliferation culture medium therein is pressed: the formula of MS+NAA0.45mg/L+ sucrose 35g/L+ agar 10g/L is prepared, and pH is set
It is set to 6.4.
4), strengthening seedling and rooting: the tufted seedling that step 3) Multiplying culture is obtained, choose growing way it is vigorous, uniform, long to 2~
Tufted seedling is split from base portion and is expanded for the second time numerous by 3cm, and coefficient is set as 8 times, is uniformly transferred to strengthening seedling and rooting culture
Strengthening seedling and rooting culture is carried out on base；
The condition of culture being provided with are as follows: strong sprout initial stage 15 days, carry out dark culture, temperature setting is 25 DEG C, humidity 70%；Later period
Setting 62 days carries out illumination and dark culture alternate culture, and temperature setting is 24~26 DEG C, light application time 12h/d, intensity of illumination
For 3000lux, 12 hours dark cultures；Culture terminates to obtain strengthening seedling and rooting；
Strengthening seedling and rooting culture medium therein is pressed: MS+NAA0.35mg/L+ sucrose 35g/L+ agar 10g/L+ banana 120g/L's matches
Side is prepared, and pH is set as 6.3.
5), transplant: the seedling obtained according to step 3) strengthening seedling and rooting culture is grown to 4~5cm high, major part substantially to be grown
When length is more than or equal to root at least 2 of 2cm, acquisition can bottle outlet plant seedling, can bottle outlet plantation seedling taken from tissue culture bottle
Out, root is removed using clear water and carries culture medium, can be obtained plantation seedling.
Potash fertilizer 0.3g/L, siliceous fertilizer 0.2g/L are also added in MS culture medium therein.
Wherein the basic used time of embodiment 2 reaches 94% in the plantation seedling planting percent that 204 day time completed to obtain, residue kind
Seedling also complies with requirement after carrying out a few days culture again, and planting percent is 99% in total, and whole seedling leaf color is bright and new beautiful, vertically
Tall and straight, growth curvature phenomenon is unobvious, and resulting single seedling weight is generally more than the seedling in embodiment 1.
Comparative example 1:
Compared with Example 1, the time of the setting condition of culture mid-early stage in step 2) to step 4) does not use dark treatment,
Directly whole process is cultivated by the way of illumination and dark culture alternate culture, other are same as Example 1.
Protocorm was obtained within the used time 66 days in total in step 2), in step 3) used time 78 days in total, in step 4) used time in total
90 days, 234 days total used times are added up to complete the seedling of 84% seedling, the seedling of subsequent completion 94%, and the seedling in comparative example 1 is curved
Qu Xianxiang presence is more obvious, and leaf color is more dim, and seedling is ineffective.
Comparative example 2:
Compared with Example 2, the time of the setting condition of culture mid-early stage in step 2) to step 4) does not use dark treatment,
Directly whole process is cultivated by the way of illumination and dark culture alternate culture, other are same as Example 2.
Protocorm was obtained within the used time 67 days in total in step 2), in step 3) used time 82 days in total, in step 4) used time in total
92 days, 241 days total used times are added up to complete the seedling of 85% seedling, the seedling of subsequent completion 95%, and the seedling in comparative example 2 is curved
Qu Xianxiang presence is more obvious, and leaf color is more dim, and seedling is ineffective, and the seedling quality of relative contrast's example 1 is mentioned
It rises.
As can be seen from the above results, the present invention connects by using the progress early period dark culture in each Multiplying culture step
The mode of nearly two week, can effectively shorten the growth time of seedlings of Dendrobium officinale, and whole planting percent is higher, and kind is planted seedlings
Condition it is more preferable, more market value, in addition adding potash fertilizer and siliceous fertilizer in the medium also can effectively promote strong sprout Growing Quality.
Candidum tissue culturing rapid propagation method disclosed by the invention, the tissue-cultured seedling cultivated is healthy and strong, transplanting survival rate
Height, quality are stablized, may advantageously facilitate the production-scale expansion of dendrobium candidum, have a vast market foreground.It is disclosed by the invention
Candidum tissue culturing rapid propagation method, is bred using tissue culture technology, and proliferation efficiency is very high, can be realized iron sheet stone
The large-scale production of dry measure used in former times, it is easy to operate, it is easy to spread.
In addition, it should be understood that although this specification is described in terms of embodiments, but not each embodiment is only wrapped
Containing an independent technical solution, this description of the specification is merely for the sake of clarity, and those skilled in the art should
It considers the specification as a whole, the technical solutions in the various embodiments may also be suitably combined, forms those skilled in the art
The other embodiments being understood that.
Claims (5)
1. a kind of candidum tissue culturing rapid propagation method, it is characterised in that: successively the following steps are included: 1), materials: select life
Knot 90 days or more uncracked mature capsules, are used sterile washing lotion to clean, drain standby on long healthy and strong dendrobium candidum
With；
2), seed induction sprout and differentiation: by the resulting capsule of step 1) it is sterilized after, cut off each 0 .3 in its head and the tail both ends~
It is longitudinally splitted after 0 .7cm, takes out seed, seed is uniformly inoculated into seed and is sprouted on differential medium, is carried out in tissue culture room
Culture；
Condition of culture is set are as follows: induction initial stage 10~15 days carries out dark culture, and temperature setting is 24~26 DEG C, humidity 60~
70%；Later period 43~47 days, illumination and dark culture alternate culture are carried out, temperature setting is 24~26 DEG C, humidity 60~70%, light
It is 8h/d, intensity of illumination 1000-1200lux, 16 hours dark cultures according to the time；Culture terminates to obtain protocorm；
3), the proliferation of seedling: step 2) protocorm obtained will be chosen that growing way is vigorous, uniform protocorm, carried out primary
Expand numerous, coefficient is set as 95~105 times, and uniformly switching carries out Multiplying culture in seedling proliferation culture medium；
Condition of culture is set are as follows: proliferation initial stage 10~15 days carries out dark culture, and temperature setting is 24~26 DEG C, humidity 60~
70%；Later period 45~75 days, illumination and dark culture alternate culture are carried out, temperature setting is 24~26 DEG C, humidity 60~70%, light
It is 10h/d, intensity of illumination 2000-2500lux, 14 hours dark cultures according to the time；Culture terminates to obtain tufted seedling；
4), strengthening seedling and rooting: the tufted seedling that step 3) Multiplying culture is obtained, selection growing way is vigorous, uniform, long to 2~3cm, will
Tufted seedling is split from base portion to be expanded numerous for the second time, and coefficient is set as 7-~10 times, is uniformly transferred to strengthening seedling and rooting culture
Strengthening seedling and rooting culture is carried out on base；
Condition of culture is set are as follows: strong sprout initial stage 13~15 days, dark culture is carried out, temperature setting is 24~26 DEG C, humidity 60~
70%；Later period 65~75 days, illumination and dark culture alternate culture are carried out, temperature setting is 24~26 DEG C, light application time 12h/
D, intensity of illumination 2800-3000lux, 12 hours dark cultures；Culture terminates to obtain strengthening seedling and rooting；
5), transplanting: the seedling length to strengthening seedling and rooting culture acquisition to 4~5cm high and grows root of the length more than or equal to 2cm extremely
At few 3, acquisition can bottle outlet plantation seedling will, then pass through the outdoor hardening of two weeks to one month, tissue-cultured seedling allowed to fit in bottle
After answering natural environment, bottle plantation seedling takes out from tissue culture bottle, removes root using clear water and carries culture medium, can be obtained strong
Strong, high survival rate plantation seedling.
2. a kind of candidum tissue culturing rapid propagation method according to claim 1, it is characterised in that: in the step 2)
Seed sprout differential medium are as follows: MS+NAA0.25~0.35mg/L+ sucrose 25~35g/L+, 5~10g/L+ of agar potato 15
~20g/L, pH 6.3.
3. a kind of candidum tissue culturing rapid propagation method according to claim 1, it is characterised in that: in the step 3)
Seedling proliferation culture medium are as follows: MS+NAA0.45~0.5mg/L+ sucrose 25~35g/L+ agar 5~10g/L, pH be 6.4~
6.5。
4. a kind of candidum tissue culturing rapid propagation method according to claim 1, it is characterised in that: in the step 4)
Strengthening seedling and rooting culture medium are as follows: 110~120g/ of MS+NAA0.35mg/L+ sucrose 25~35g/L+ agar 5~10g/L+ banana
L, pH are 6.3~6.5.
5. according to a kind of any candidum tissue culturing rapid propagation method of claim 2-4, it is characterised in that: the MS
0.35~0.45g/L of potash fertilizer, 0.15~0.25g/L of siliceous fertilizer are also added in culture medium.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116171855A (en) * | 2023-02-09 | 2023-05-30 | 云南山里红生物科技有限公司 | Dendrobium officinale tissue culture method and culture medium formula thereof |
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