CN109105265A - A kind of stem section induction Multiple Buds rapid propagation method of dendrobium candidum - Google Patents
A kind of stem section induction Multiple Buds rapid propagation method of dendrobium candidum Download PDFInfo
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- CN109105265A CN109105265A CN201811333144.3A CN201811333144A CN109105265A CN 109105265 A CN109105265 A CN 109105265A CN 201811333144 A CN201811333144 A CN 201811333144A CN 109105265 A CN109105265 A CN 109105265A
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- stem section
- dendrobium candidum
- multiple buds
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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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- 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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Abstract
The invention discloses a kind of stem sections of dendrobium candidum to induce Multiple Buds rapid propagation method, the following steps are included: Step 1: stem section explant is proliferated: taking that annual dendrobium candidum is sturdy, good plant, blade is cut with sharp scalpel, it is rinsed with flowing water, soft bristle tooth brush cleans down stem section, the crust for gently peelling off stem section white does explant with stem section；It is impregnated 30 seconds, aseptic water washing 3~4 times with 70~75% alcohol again；With white water treatment 10 minutes of 10 times, aseptic water washing 3~4 times；Then it is impregnated and is shaken 10~15 minutes with 0.1% mercuric chloride solution, using the stem section of dendrobium candidum defect individual as explant, establish a set of more complete stem section induction Multiple Buds tissue training rapid propagation system, stem section clone technology has germination early, rootage duration is short, and reproduction speed is fast, is not limited by geographical environment, weather conditions, not by the season limit of acquisition seed, it can be achieved that anniversary clone's production.
Description
Technical field
The invention belongs to dendrobium candidum quick breeding technology fields, and in particular to a kind of stem section induction of dendrobium candidum is grown thickly
Bud rapid propagation method.
Background technique
Dendrobium candidum, nickname iron sheet orchid, ribbed hedyotis herb, are the perennial herbaceous plant that grows nonparasitically upon another plant of orchid family Dendrobium, because its stem is in
Iron green, so dendrobium candidum of gaining the name.Dendrobium candidum is had long enjoyed a good reputation with its high medical value, have eliminating the phlegm, it is anti-oxidant, mention
High body's immunity, hypoglycemic and anticancer and other effects.Dendrobium nobile product --- the Tiepi Fengdou to circulate currently on the market, be by
Dendrobium candidum after working process is economic value and the high rare Chinese herbal medicine of medical value simply.Dendrobium candidum is dendrobium nobile
Superfine product in platymiscium, it gains the name because epidermis is green in iron.Since thousands of years always with the quilts such as ginseng, ganoderma lucidum, cordyceps sinensis
It is classified as top grade Chinese medicine, is known as in the Tang Dynasty medicine classical " Taoist Scriptures " " first of Chinese nine big mesonas ".Because its growth cycle is slow,
Extremely difficult stock, wild resource is endangered, and the rare and endangered medicinal plant of focused protection is classified as within 1987 by country.
Dendrobium candidum is the superfine product in dendrobium nobile, has unique nourishing Yin and promoting production of body fluid effect, by ancient Chinese medicine doctor and medicine ancient books and records
High praise.The version Pharmacopoeias of the People's Republic of China in 2010 of implementation have increased the single-row mark of dendrobium candidum newly from October 1st, 2010
Standard, having rewritten dendrobium candidum does not have the history of national standard.In new edition pharmacopeia, increase dendrobium candidum indentification by TLC,
Mannose and glucose peaks area when inspection items such as impurity, moisture, total ash, extract, and formulated polysaccharide and sweet dew
Sugared content measuring method makes the identification with specificity and quality controllable content assaying method.New standard can not only
Enough detect the true and false of dendrobium candidum, moreover it is possible to the further quality of control and identification dendrobium candidum.It is obtained by tissue cultures
The approach for obtaining dendrobium candidum regeneration plant mainly has: with carrying out after mature seed disinfection, aseptic seeding generates seedling or formation is former
Bulb is simultaneously proliferated, and then differentiation and development is at intact plant, is suitable for that explant (stem section, stem apex, young shoot, blade etc.) is formed with inducing
Intend protocorm or Multiple Buds to realize quick breeding, to obtain a large amount of regeneration plants.
Under natural conditions, for dendrobium candidum because seed lacks endosperm, fertility is extremely low, along with the mankind are for a long time without section
Digging processed, wild resource is increasingly exhausted, the dendrobium candidum to circulate currently on the market, is substantially artificial cultivation kind, but it is produced
Amount is not able to satisfy medicinal demand now still, in order to protect existing Wild ornamental resources, keeps the provenance of dendrobium candidum to stablize and supplies
It answers, increasing to the fast numerous research of candidum tissue culturing, the present invention directly induces Multiple Buds using stem section, does not need by luring
Protocorm differentiation adventitious bud is led, the good characteristic of hereditary female parent can be stablized, then reduces the probability that offspring morphs.
Summary of the invention
The technical problem to be solved by the present invention is to overcome the existing defects, and the stem section induction for providing a kind of dendrobium candidum is grown thickly
Bud rapid propagation method, to solve under natural conditions mentioned above in the background art, dendrobium candidum is numerous because seed lacks endosperm
It is extremely low to grow ability, adds the mankind's uncontrolled digging for a long time, wild resource is increasingly exhausted, the iron sheet to circulate currently on the market
Dendrobium nobile is substantially artificial cultivation kind, but its yield is not able to satisfy medicinal demand now still, in order to protect existing wild species
Matter resource keeps the provenance of dendrobium candidum to stablize supply, numerous research fast to candidum tissue culturing it is increasing the problem of.
To achieve the above object, the invention provides the following technical scheme: a kind of stem section induction Multiple Buds of dendrobium candidum are fast
Fast propagation method, comprising the following steps:
Step 1: stem section explant is proliferated: taking that annual dendrobium candidum is sturdy, good plant, with sharp scalpel
Blade is cut, is rinsed with flowing water, soft bristle tooth brush cleans down stem section, gently peels off the crust of stem section white, does explant with stem section
Body；It is impregnated 30 seconds, aseptic water washing 3~4 times with 70~75% alcohol again；It is sterile with white water treatment 10 minutes of 10 times
Water rinses 3~4 times；Then it is impregnated and is shaken 10~15 minutes with 0.1% mercuric chloride solution, with aseptic water washing 3~4 times, dried in the air
Solid carbon dioxide point, cuts stem section both ends, each stem section 1.5cm or so, 2 stipes of band, every bottle of culture medium inoculated one with sterile scalpel
A stem section, avoids cross contamination, and the stem section after disinfection is inoculated into Primary culture base: spend No. 1+BA1~5mg/L+KT1 of treasured~
5mg/L+NAA0.1~2mg/L+ potato juice 5000~15000mg/L+ coconut palm cream 10% (V/V), dark culture one week, then one
Lamp culture, daily illumination 12 hours, 25 ± 2 DEG C of temperature.By culture in 3 months, explant starting success rate reached 70%, one
There are 2~3 budlets to grow in a stem section.
Step 2: Multiple Buds strong sprout: the Multiple Buds that stem section is induced are inoculated into culture medium: spending precious No. 1+potato juice
5000~15000mg/L+ banana 5000~15000mg/L+ active carbon 1g/L, 2 lamp cultures, 25 ± 2 DEG C of temperature, daily illumination
After 12 hours, 3 months, seedling grows to 7cm, may be used as female bottle switchback proliferation.
Step 3: stem section is proliferated: the leaf of dendrobium candidum and root are cut, each 2 section of stem section band lies in culture medium:
Spend No. 1+BA1~5mg/L+KT1~5mg/L+NAA0.1 of treasured~2mg/L+ potato juice 5000~15000mg/L+ coconut palm cream 10%
(V/V) a lamp culture, daily illumination 12 hours, 25 ± 2 DEG C of temperature.By culture in 3 months, there are 3~4 in a stem section
Budlet grows.
Step 4: Multiple Buds are taken root: the proliferation clump bud that stem section is induced, be inoculated into spend precious No. 1+potato juice 5000~
15000mg/L+ banana 5000~15000mg/L+ active carbon 1g/L, 2 lamp cultures, 25 ± 2 DEG C of temperature, daily illumination 12 is small
When, after 4 months, seedling grows to 10cm, there is 8~10 leaves, and a young plant has 3~4 roots, 3~8cm of root long, rooting rate up to 99%,
Bottle seedling uniformity reaches 95%, the bottle seedling that then will be taken root, lower to wash seed plant to greenhouse refining, bottle outlet.
Preferably, adding a drop dish washing liquid in the step 1 in 10 times of bleaching water is spreader-sticker.
Preferably, adding a drop dish washing liquid in the step 1 in 0.1% mercuric chloride solution is spreader-sticker.
Preferably, sterile water is the cooling sterile water by high temperature sterilization production after cold water heat is opened in the step 1.
Preferably, sterile scalpel is made in the step 1 of stainless steel material.
Preferably, lamp is LED plant growth lamp in the step 2.
Preferably, BA is 6- benzyl aminoadenine in the step 3.
Preferably, NAA is methyl α-naphthyl acetate in the step 3.
Preferably, KT is kinetin in the step 3.
Compared with prior art, the present invention provides a kind of stem sections of dendrobium candidum to induce Multiple Buds rapid propagation method,
Have it is following the utility model has the advantages that
1, the present invention establishes a set of more complete stem section induced bundle using the stem section of dendrobium candidum defect individual as explant
Tissue of sprouting trains rapid propagation system, and stem section clone technology has germination early, and rootage duration is short, and reproduction speed is fast, not by geographical environment,
Weather conditions limitation, not by the season limit of acquisition seed, it can be achieved that anniversary clone's production.
2, dendrobium candidum of the present invention generally uses protocorm approach, Protocorm Multiplication, protocorm differentiation, multistep of taking root stream
Journey；From cell to micro- seedling, seedling, strong sprout, the period about 1 year, present invention proliferation, two steps of taking root, using two kinds of culture mediums,
Simplify production routine, can emerge from stem section induced bud to taking root bottle outlet 7 months.
3, the present invention is maintained maternal characteristic, subtracted by the optimization of cutting method, culture medium prescription using using buds to propagate buds approach
The variation of few dendrobium candidum, the uniformity for improving bottle seedling, make the uniformity of bottle seedling reach 95%.
It is not directed to part in the device to be the same as those in the prior art or can be realized by using the prior art, structure of the invention
It is scientific and reasonable, it is safe and convenient to use, it provides a great help for people.
Detailed description of the invention
Attached drawing is used to provide further understanding of the present invention, and constitutes part of specification, with reality of the invention
It applies example to be used to explain the present invention together, not be construed as limiting the invention, in the accompanying drawings:
Fig. 1 is that a kind of stem section of dendrobium candidum proposed by the present invention induces Multiple Buds rapid propagation method Technology Roadmap.
Specific embodiment
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Site preparation description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts every other
Embodiment shall fall within the protection scope of the present invention.
Referring to Fig. 1, the present invention provides a kind of technical solution: a kind of stem section induction Multiple Buds of dendrobium candidum are quickly bred
Method, comprising the following steps:
Step 1: stem section explant is proliferated: taking that annual dendrobium candidum is sturdy, good plant, with sharp scalpel
Blade is cut, is rinsed with flowing water, soft bristle tooth brush cleans down stem section, gently peels off the crust of stem section white, does explant with stem section
Body；It is impregnated 30 seconds, aseptic water washing 3~4 times with 70~75% alcohol again；It is sterile with white water treatment 10 minutes of 10 times
Water rinses 3~4 times；Then it is impregnated and is shaken 10~15 minutes with 0.1% mercuric chloride solution, with aseptic water washing 3~4 times, dried in the air
Solid carbon dioxide point, cuts stem section both ends, each stem section 1.5cm or so, 2 stipes of band, every bottle of culture medium inoculated one with sterile scalpel
A stem section, avoids cross contamination, and the stem section after disinfection is inoculated into Primary culture base: spend No. 1+BA1~5mg/L+KT1 of treasured~
5mg/L+NAA0.1~2mg/L+ potato juice 5000~15000mg/L+ coconut palm cream 10% (V/V), dark culture one week, then one
Lamp culture, daily illumination 12 hours, 25 ± 2 DEG C of temperature.By culture in 3 months, explant starting success rate reached 70%, one
There are 2~3 budlets to grow in a stem section.
Step 2: Multiple Buds strong sprout: the Multiple Buds that stem section is induced are inoculated into culture medium: spending precious No. 1+potato juice
5000~15000mg/L+ banana 5000~15000mg/L+ active carbon 1g/L, 2 lamp cultures, 25 ± 2 DEG C of temperature, daily illumination
After 12 hours, 3 months, seedling grows to 7cm, may be used as female bottle switchback proliferation.
Step 3: stem section is proliferated: the leaf of dendrobium candidum and root are cut, each 2 section of stem section band lies in culture medium:
Spend No. 1+BA1~5mg/L+KT1~5mg/L+NAA0.1 of treasured~2mg/L+ potato juice 5000~15000mg/L+ coconut palm cream 10%
(V/V) a lamp culture, daily illumination 12 hours, 25 ± 2 DEG C of temperature.By culture in 3 months, there are 3~4 in a stem section
Budlet grows.
Step 4: Multiple Buds are taken root: the proliferation clump bud that stem section is induced, be inoculated into spend precious No. 1+potato juice 5000~
15000mg/L+ banana 5000~15000mg/L+ active carbon 1g/L, 2 lamp cultures, 25 ± 2 DEG C of temperature, daily illumination 12 is small
When, after 4 months, seedling grows to 10cm, there is 8~10 leaves, and a young plant has 3~4 roots, 3~8cm of root long, rooting rate up to 99%,
Bottle seedling uniformity reaches 95%, the bottle seedling that then will be taken root, lower to wash seed plant to greenhouse refining, bottle outlet.
In the present invention, it is preferred that adding a drop dish washing liquid in step 1 in 10 times of bleaching water is spreader-sticker.
In the present invention, it is preferred that adding a drop dish washing liquid in step 1 in 0.1% mercuric chloride solution is spreader-sticker.
In the present invention, it is preferred that sterile water is cooling by the sterile of high temperature sterilization production after cold water heat is opened in step 1
Water.
In the present invention, it is preferred that sterile scalpel is made in step 1 of stainless steel material.
In the present invention, it is preferred that lamp is LED plant growth lamp in step 2.
In the present invention, it is preferred that BA is 6- benzyl aminoadenine in step 3.
In the present invention, it is preferred that NAA is methyl α-naphthyl acetate in step 3.
In the present invention, it is preferred that KT is kinetin in step 3.
Embodiment one
A kind of stem section induction Multiple Buds rapid propagation method of dendrobium candidum, comprising the following steps:
Step 1: stem section explant is proliferated: taking that annual dendrobium candidum is sturdy, good plant, with sharp scalpel
Blade is cut, is rinsed with flowing water, soft bristle tooth brush cleans down stem section, gently peels off the crust of stem section white, does explant with stem section
Body；It is impregnated 30 seconds, aseptic water washing 3~4 times with 70~75% alcohol again；With white water treatment 5 minutes of 10 times, sterile water
It rinses 1~3 time；Then it is impregnated and is shaken 5~10 minutes with 0.1% mercuric chloride solution, with aseptic water washing 1~3 time, dry water
Point, stem section both ends, each stem section 1.5cm or so, 2 stipes of band, one stem of every bottle of culture medium inoculated are cut with sterile scalpel
Section, avoids cross contamination, the stem section after disinfection is inoculated into Primary culture base: spending No. 1+BA1~5mg/L+KT1~5mg/L+ of treasured
NAA0.1~2mg/L+ potato juice 5000~8000mg/L+ coconut palm cream 5% (V/V), dark culture one week, a then lamp culture,
Daily illumination 12 hours, 25 ± 2 DEG C of temperature.By culture in 3 months, explant starting success rate reached 70%, a stem section
On there are 2~3 budlets to grow.
Step 2: Multiple Buds strong sprout: the Multiple Buds that stem section is induced are inoculated into culture medium: spending precious No. 1+potato juice
5000~8000mg/L+ banana 5000~8000mg/L+ active carbon 0.9g/L, 2 lamp cultures, 25 ± 2 DEG C of temperature, daily illumination
After 12 hours, 3 months, seedling grows to 7cm, may be used as female bottle switchback proliferation.
Step 3: stem section is proliferated: the leaf of dendrobium candidum and root are cut, each 2 section of stem section band lies in culture medium:
Spend No. 1+BA1~5mg/L+KT1~5mg/L+NAA0.1 of treasured~5% (V/ of 2mg/L+ potato juice 5000~8000mg/L+ coconut palm cream
V) lamp a culture, daily illumination 12 hours, 25 ± 2 DEG C of temperature.By culture in 3 months, have in a stem section 3~4 small
Bud is grown.
Step 4: Multiple Buds are taken root: the proliferation clump bud that stem section is induced, be inoculated into spend precious No. 1+potato juice 5000~
8000mg/L+ banana 5000~8000mg/L+ active carbon 0.9g/L, 2 lamp cultures, 25 ± 2 DEG C of temperature, daily illumination 12 is small
When, after 4 months, seedling grows to 10cm, there is 8~10 leaves, and a young plant has 3~4 roots, 3~8cm of root long, rooting rate up to 85%,
Bottle seedling uniformity reaches 95%, the bottle seedling that then will be taken root, lower to wash seed plant to greenhouse refining, bottle outlet.
In the present invention, it is preferred that adding a drop dish washing liquid in step 1 in 10 times of bleaching water is spreader-sticker.
In the present invention, it is preferred that adding a drop dish washing liquid in step 1 in 0.1% mercuric chloride solution is spreader-sticker.
In the present invention, it is preferred that sterile water is cooling by the sterile of high temperature sterilization production after cold water heat is opened in step 1
Water.
In the present invention, it is preferred that sterile scalpel is made in step 1 of stainless steel material.
In the present invention, it is preferred that lamp is LED plant growth lamp in step 2.
In the present invention, it is preferred that BA is 6- benzyl aminoadenine in step 3.
In the present invention, it is preferred that NAA is methyl α-naphthyl acetate in step 3.
In the present invention, it is preferred that KT is kinetin in step 3.
Embodiment two
A kind of stem section induction Multiple Buds rapid propagation method of dendrobium candidum, comprising the following steps:
Step 1: stem section explant is proliferated: taking that annual dendrobium candidum is sturdy, good plant, with sharp scalpel
Blade is cut, is rinsed with flowing water, soft bristle tooth brush cleans down stem section, gently peels off the crust of stem section white, does explant with stem section
Body；It is impregnated 30 seconds, aseptic water washing 3~4 times with 70~75% alcohol again；It is sterile with white water treatment 10 minutes of 10 times
Water rinses 3~4 times；Then it is impregnated and is shaken 10~15 minutes with 0.1% mercuric chloride solution, with aseptic water washing 3~4 times, dried in the air
Solid carbon dioxide point, cuts stem section both ends, each stem section 1.5cm or so, 2 stipes of band, every bottle of culture medium inoculated one with sterile scalpel
A stem section, avoids cross contamination, and the stem section after disinfection is inoculated into Primary culture base: spend No. 1+BA1~5mg/L+KT1 of treasured~
5mg/L+NAA0.1~2mg/L+ potato juice 5000~15000mg/L+ coconut palm cream 10% (V/V), dark culture one week, then one
Lamp culture, daily illumination 12 hours, 25 ± 2 DEG C of temperature.By culture in 3 months, explant starting success rate reached 70%, one
There are 2~3 budlets to grow in a stem section.
Step 2: Multiple Buds strong sprout: the Multiple Buds that stem section is induced are inoculated into culture medium: spending precious No. 1+potato juice
5000~15000mg/L+ banana 5000~15000mg/L+ active carbon 1g/L, 2 lamp cultures, 25 ± 2 DEG C of temperature, daily illumination
After 12 hours, 3 months, seedling grows to 7cm, may be used as female bottle switchback proliferation.
Step 3: stem section is proliferated: the leaf of dendrobium candidum and root are cut, each 2 section of stem section band lies in culture medium:
Spend No. 1+BA1~5mg/L+KT1~5mg/L+NAA0.1 of treasured~2mg/L+ potato juice 5000~15000mg/L+ coconut palm cream 10%
(V/V) a lamp culture, daily illumination 12 hours, 25 ± 2 DEG C of temperature.By culture in 3 months, there are 3~4 in a stem section
Budlet grows.
Step 4: Multiple Buds are taken root: the proliferation clump bud that stem section is induced, be inoculated into spend precious No. 1+potato juice 5000~
15000mg/L+ banana 5000~15000mg/L+ active carbon 1g/L, 2 lamp cultures, 25 ± 2 DEG C of temperature, daily illumination 12 is small
When, after 4 months, seedling grows to 10cm, there is 8~10 leaves, and a young plant has 3~4 roots, 3~8cm of root long, rooting rate up to 99%,
Bottle seedling uniformity reaches 95%, the bottle seedling that then will be taken root, lower to wash seed plant to greenhouse refining, bottle outlet.
In the present invention, it is preferred that adding a drop dish washing liquid in step 1 in 10 times of bleaching water is spreader-sticker.
In the present invention, it is preferred that adding a drop dish washing liquid in step 1 in 0.1% mercuric chloride solution is spreader-sticker.
In the present invention, it is preferred that sterile water is cooling by the sterile of high temperature sterilization production after cold water heat is opened in step 1
Water.
In the present invention, it is preferred that sterile scalpel is made in step 1 of stainless steel material.
In the present invention, it is preferred that lamp is LED plant growth lamp in step 2.
In the present invention, it is preferred that BA is 6- benzyl aminoadenine in step 3.
In the present invention, it is preferred that NAA is methyl α-naphthyl acetate in step 3.
In the present invention, it is preferred that KT is kinetin in step 3.
Embodiment three
A kind of stem section induction Multiple Buds rapid propagation method of dendrobium candidum, comprising the following steps:
Step 1: stem section explant is proliferated: taking that annual dendrobium candidum is sturdy, good plant, with sharp scalpel
Blade is cut, is rinsed with flowing water, soft bristle tooth brush cleans down stem section, gently peels off the crust of stem section white, does explant with stem section
Body；It is impregnated 30 seconds, aseptic water washing 3~4 times with 70~75% alcohol again；It is sterile with white water treatment 15 minutes of 10 times
Water rinses 4~5 times；Then it is impregnated and is shaken 15~20 minutes with 0.1% mercuric chloride solution, with aseptic water washing 4~5 times, dried in the air
Solid carbon dioxide point, cuts stem section both ends, each stem section 1.5cm or so, 2 stipes of band, every bottle of culture medium inoculated one with sterile scalpel
A stem section, avoids cross contamination, and the stem section after disinfection is inoculated into Primary culture base: spend No. 1+BA1~5mg/L+KT1 of treasured~
5mg/L+NAA0.1~2mg/L+ potato juice 11000~15000mg/L+ coconut palm cream 15% (V/V), dark culture one week, then one
Branch lamp culture, daily illumination 12 hours, 25 ± 2 DEG C of temperature.By culture in 3 months, explant starting success rate reached 70%,
There are 2~3 budlets to grow in one stem section.
Step 2: Multiple Buds strong sprout: the Multiple Buds that stem section is induced are inoculated into culture medium: spending precious No. 1+potato juice
11000~15000mg/L+ banana 11000~15000mg/L+ active carbon 1.1g/L, 2 lamp cultures, 25 ± 2 DEG C of temperature, daily
After illumination 12 hours, 3 months, seedling grows to 7cm, may be used as female bottle switchback proliferation.
Step 3: stem section is proliferated: the leaf of dendrobium candidum and root are cut, each 2 section of stem section band lies in culture medium:
Spend No. 1+BA1~5mg/L+KT1~5mg/L+NAA0.1 of treasured~2mg/L+ potato juice 11000~15000mg/L+ coconut palm cream 15%
(V/V) a lamp culture, daily illumination 12 hours, 25 ± 2 DEG C of temperature.By culture in 3 months, there are 3~4 in a stem section
Budlet grows.
Step 4: Multiple Buds are taken root: the proliferation clump bud that stem section is induced, be inoculated into spend precious No. 1+potato juice 11000~
15000mg/L+ banana 11000~15000mg/L+ active carbon 1.1g/L, 2 lamp cultures, 25 ± 2 DEG C of temperature, daily illumination 12
Hour, after 4 months, seedling grows to 10cm, there is 8~10 leaves, and a young plant has 3~4 roots, 3~8cm of root long, and rooting rate reaches
95%, bottle seedling uniformity reaches 95%, the bottle seedling that then will be taken root, lower to wash seed plant to greenhouse refining, bottle outlet.
In the present invention, it is preferred that adding a drop dish washing liquid in step 1 in 10 times of bleaching water is spreader-sticker.
In the present invention, it is preferred that adding a drop dish washing liquid in step 1 in 0.1% mercuric chloride solution is spreader-sticker.
In the present invention, it is preferred that sterile water is cooling by the sterile of high temperature sterilization production after cold water heat is opened in step 1
Water.
In the present invention, it is preferred that sterile scalpel is made in step 1 of stainless steel material.
In the present invention, it is preferred that lamp is LED plant growth lamp in step 2.
In the present invention, it is preferred that BA is 6- benzyl aminoadenine in step 3.
In the present invention, it is preferred that NAA is methyl α-naphthyl acetate in step 3.
In the present invention, it is preferred that KT is kinetin in step 3.
The present invention establishes a set of more complete stem section induction and grows thickly using the stem section of dendrobium candidum defect individual as explant
Bud tissue trains rapid propagation system, and stem section clone technology has germination early, and rootage duration is short, and reproduction speed is fast, not by geographical environment, gas
Time condition limitation, not by the season limit of acquisition seed, it can be achieved that anniversary clone's production.
Dendrobium candidum of the present invention generally uses protocorm approach, Protocorm Multiplication, protocorm differentiation, multistep of taking root process；
From cell to micro- seedling, seedling, strong sprout, the period about 1 year, present invention proliferation, two steps of taking root used two kinds of culture mediums, letter
Change production routine, can emerge from stem section induced bud to taking root bottle outlet 7 months.
The present invention maintains maternal characteristic using using buds to propagate buds approach by the optimization of cutting method, culture medium prescription, reduces
The variation of dendrobium candidum, the uniformity for improving bottle seedling, make the uniformity of bottle seedling reach 95%.
It is not directed to part in the device to be the same as those in the prior art or can be realized by using the prior art, structure of the invention
It is scientific and reasonable, it is safe and convenient to use, it provides a great help for people.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
A variety of variations, modification, replacement can be carried out to these embodiments without departing from the principles and spirit of the present invention by understanding
And modification, the scope of the present invention is defined by the appended.
Claims (9)
1. a kind of stem section of dendrobium candidum induces Multiple Buds rapid propagation method, which comprises the following steps:
Step 1: stem section explant is proliferated: taking that annual dendrobium candidum is sturdy, good plant, cut with sharp scalpel
Blade is rinsed with flowing water, and soft bristle tooth brush cleans down stem section, gently peels off the crust of stem section white, does explant with stem section；Again
It is impregnated 30 seconds, aseptic water washing 3~4 times with 70~75% alcohol；With white water treatment 10 minutes of 10 times, aseptic water washing
3~4 times；Then it is impregnated and is shaken 10~15 minutes with 0.1% mercuric chloride solution, with aseptic water washing 3~4 times, dry moisture,
Stem section both ends, each stem section 1.5cm or so are cut with sterile scalpel, 2 stipes of band, one stem section of every bottle of culture medium inoculated,
Cross contamination is avoided, the stem section after disinfection is inoculated into Primary culture base: spending No. 1+BA1~5mg/L+KT1~5mg/L+ of treasured
NAA0.1~2mg/L+ potato juice 5000~15000mg/L+ coconut palm cream 10% (V/V), dark culture one week, a then lamp training
It supports, daily illumination 12 hours, 25 ± 2 DEG C of temperature.By culture in 3 months, explant starting success rate reached 70%, a stem
There are 2~3 budlets to grow in section.
Step 2: Multiple Buds strong sprout: the Multiple Buds that stem section is induced are inoculated into culture medium: spend precious No. 1+potato juice 5000~
15000mg/L+ banana 5000~15000mg/L+ active carbon 1g/L, 2 lamp cultures, 25 ± 2 DEG C of temperature, daily illumination 12 is small
When, after 3 months, seedling grows to 7cm, may be used as female bottle switchback proliferation.
Step 3: stem section is proliferated: the leaf of dendrobium candidum and root being cut, each 2 section of stem section band lies in culture medium: spending treasured 1
Number+BA1~5mg/L+KT1~5mg/L+NAA0.1~2mg/L+ potato juice 5000~15000mg/L+ coconut palm cream 10% (V/V)
One lamp culture, daily illumination 12 hours, 25 ± 2 DEG C of temperature.By culture in 3 months, there are 3~4 budlets in a stem section
It grows.
Step 4: Multiple Buds are taken root: the proliferation clump bud that stem section is induced, be inoculated into spend precious No. 1+potato juice 5000~
15000mg/L+ banana 5000~15000mg/L+ active carbon 1g/L, 2 lamp cultures, 25 ± 2 DEG C of temperature, daily illumination 12 is small
When, after 4 months, seedling grows to 10cm, there is 8~10 leaves, and a young plant has 3~4 roots, 3~8cm of root long, rooting rate up to 99%,
Bottle seedling uniformity reaches 95%, the bottle seedling that then will be taken root, lower to wash seed plant to greenhouse refining, bottle outlet.
2. a kind of stem section of dendrobium candidum according to claim 1 induces Multiple Buds rapid propagation method, it is characterised in that:
Adding a drop dish washing liquid in the step 1 in 10 times of bleaching water is spreader-sticker.
3. a kind of stem section of dendrobium candidum according to claim 1 induces Multiple Buds rapid propagation method, it is characterised in that:
Adding a drop dish washing liquid in the step 1 in 0.1% mercuric chloride solution is spreader-sticker.
4. a kind of stem section of dendrobium candidum according to claim 1 induces Multiple Buds rapid propagation method, it is characterised in that:
Sterile water is the cooling sterile water by high temperature sterilization production after cold water heat is opened in the step 1.
5. a kind of stem section of dendrobium candidum according to claim 1 induces Multiple Buds rapid propagation method, it is characterised in that:
Sterile scalpel is made in the step 1 of stainless steel material.
6. a kind of stem section of dendrobium candidum according to claim 1 induces Multiple Buds rapid propagation method, it is characterised in that:
Lamp is LED plant growth lamp in the step 2.
7. a kind of stem section of dendrobium candidum according to claim 1 induces Multiple Buds rapid propagation method, it is characterised in that:
BA is 6- benzyl aminoadenine in the step 3.
8. a kind of stem section of dendrobium candidum according to claim 1 induces Multiple Buds rapid propagation method, it is characterised in that:
NAA is methyl α-naphthyl acetate in the step 3.
9. a kind of stem section of dendrobium candidum according to claim 1 induces Multiple Buds rapid propagation method, it is characterised in that:
KT is kinetin in the step 3.
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| CN103477976A (en) * | 2013-07-12 | 2014-01-01 | 广东省农业科学院作物研究所 | Stem tissue culture seedling method of dendrobium candidum |
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| US20160143236A1 (en) * | 2013-07-08 | 2016-05-26 | South China Botanical Garden, Chinese Academy Of Sciences | Dendrobium in vitro crossbreeding method |
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| CN103053425A (en) * | 2013-01-22 | 2013-04-24 | 杨宝明 | Rapid propagation method for tissue cultivation of dendrobium candidum stem |
| US20160143236A1 (en) * | 2013-07-08 | 2016-05-26 | South China Botanical Garden, Chinese Academy Of Sciences | Dendrobium in vitro crossbreeding method |
| CN103477976A (en) * | 2013-07-12 | 2014-01-01 | 广东省农业科学院作物研究所 | Stem tissue culture seedling method of dendrobium candidum |
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