CN102972299B - Rapid propagation method for dendrobium officinale stem tissue culture - Google Patents
Rapid propagation method for dendrobium officinale stem tissue culture Download PDFInfo
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Abstract
The invention discloses a rapid propagation method for dendrobium officinale stem tissue culture, and belongs to the technical field of biology. According to the rapid propagation method, stem sections (above the third section from the root) of dendrobium officinale with the growth period more than one year are used for carrying out rapid propagation for the tissue culture. The rapid propagation method comprises the following steps of: preparation of implants except the stems, sterilization, induction culture, subculture, strong seedling culture, rooting culture and light culture hardening-seedling. The rapid propagation method for the dendrobium officinale stem tissue culture, provided by the invention, has the advantages of easiness of material obtaining, large material amount and no limitation on the seasons; the maternal biological property can be kept well; a culture technology is simple and easy to operate and according to the method, the low pollution rate and the rapid propagation speed can be realized; and the rapid propagation method for the dendrobium officinale stem tissue culture has the important meanings on resource regeneration and continuous use of scarce medicinal plants in imminent danger.
Description
Technical field
The invention belongs to biological technical field, relate to a kind of plant cultivation method, relate in particular to dendrobium candidum stem segment tissue culture fast breeding method.
Background technology
Dendrobium candidum (Dendrobium officinale Kimura et Migo), belong to the orchid family epiphytic orchid class, its medicinal material is commonly called as Tiepi Fengdou, claims again ribbed hedyotis herb, be a kind of traditional rare Chinese medicine, there is effect of nourishing Yin and clearing heat, the beneficial stomach that promotes the production of body fluid, the improving eyesight that moistens the lung and relieve the cough, wets one's whistle.Modern medicine study shows, the compositions such as the polysaccharide of dendrobium candidum, alkaloid, amino acid have the effect that improves immunologic function, suppresses thrombosis, suppresses tumour and delay senility.Because Seeds of Dendrobium Candidum is tiny; there is after ripening in embryo; when fruit maturation; the growth of embryo is no more than the ball-type phase, and seed lacks endosperm, and needs the existence of fungal component; therefore under natural conditions, reproduction rate is extremely low; add uncontrolled excavating all the year round, natural resources is day by day exhausted, is listed in special-protection-by-the-State medicinal plant, Chinese Precious, Rare, Endangered second class protection plant and the world's two class protective plants.
The method that dendrobium candidum often adopts plant division, cuttage etc. to nourish and generate aborning, but the low problem of reproduction coefficient is difficult to solve.Since the vegetative propagation technique that utilizes stem of noble dendrobium base point tissue to cultivate virus-free plant from nineteen sixty Frenchman Gmorel is founded, stem of noble dendrobium tissue culture technique is progressively deepened, and stem of noble dendrobium tissue cultivating and seedling progressively tends to industrialization especially in recent years.There are some researches show the particularity due to the stem of noble dendrobium, in the tissue of the stem of noble dendrobium is cultivated, the selection of explant is very crucial factor, and different draws materials position and period, and the result of cultivation is also different.In current research, the in vitro tissue culture method of dendrobium candidum, normally taking ripe or immature seed and tender stem apex, axillalry bud or the young stem stem section of children as explant.Although seed has the advantages such as simple to operate, survival rate is high, and quantity amplification is rapid as explant, the difficulty that explant material obtains, but become great limiting factor, and likely produce variation with the offspring of seminal propagation; Taking the tender stem apex of children, axillalry bud or young stem stem section as material, can produce very bad impact to the growth of plant again, be also subject to the restriction in season simultaneously.The present invention carries out tissue-culturing rapid propagation taking the dendrobium candidum in more than 1 year vegetative period from adult state stem sections more than root three joints as explant material, overcome the defect of above-mentioned two kinds of explant materials simultaneously, not only quantity of material is large, and all can draw materials in any season, at present there are no relevant report.
Summary of the invention
The object of the invention is to organize training explant material in vitro for dendrobium candidum and obtain difficulty, a kind of dendrobium candidum stem segment tissue culture fast breeding method is provided, the method is carried out tissue-culturing rapid propagation taking the adult state stem section of the dendrobium candidum in more than 1 year vegetative period as explant material, there is quantity of material and greatly, be not subject to seasonal restrictions advantage, can increase reproductive speed and output simultaneously, reduce group training production cost, significant to resources of medicinal plant regeneration in short supply in imminent danger and sustainable utilization.
The technical solution used in the present invention is:
A kind of dendrobium candidum stem segment tissue culture fast breeding method, comprises the following steps:
1. the preparation of stem explants: get the state dendrobium candidum stem section of growing up, remove blade, rinse well with flowing water; The dendrobium candidum that described adult state dendrobium candidum stem section is more than 1 year vegetative period is from stem sections more than root three joints;
2. the sterilization of stem explants: the stem explants cleaning up is taken out, put in beaker, mercuric chloride solution with 0.1% soaks 5 minutes, with aseptic water washing 4～5 times, put on superclean bench, to be cut into length be 1～2cm, little stem section with 2～3 joints, then soak 3 minutes with 0.1% mercuric chloride solution, with aseptic water washing 4～5 times, for subsequent use;
3. induction is cultivated: the little stem section disinfecting is inoculated in aseptic inducing culture, this culture medium prescription is: MS+6-BA0.8～1.2mg/L+NAA0.8～1.2mg/L, condition of culture: 24～26 DEG C of temperature, fluorescent lamp irradiate 24 hours/day, intensity of illumination 800～1200lx, cultivate after 40 days, stem section goes out sprouting internode director;
4. propagation is cultivated for the first time: sprouting is transferred to for the first time in proliferated culture medium, the formula of proliferated culture medium is for the first time: MS+6-BA4.8～5.2mg/L+NAA0.8～1.2mg/L, 24～26 DEG C of temperature, fluorescent lamp irradiate 24 hours/day, intensity of illumination 800～1200lx, cultivate after 30 days, grow in incision the first generation Multiple Buds that about 0.5cm is high, the propagation multiple of sprouting is more than 20 times;
5. propagation is cultivated for the second time: will after first generation Multiple Buds cutting, be transferred to for the second time in proliferated culture medium, the formula of proliferated culture medium is for the second time: MS+6-BA3.8～4.2mg/L+NAA0.8～1.2mg/L, 24～26 DEG C of temperature, fluorescent lamp irradiate 24 hours/day, intensity of illumination 800～1200lx, cultivate 30～40 days, grow in incision the second generation Multiple Buds that about 0.5cm is high, the propagation multiple of sprouting reaches more than 10 times;
6. propagation is cultivated for the third time: will after second generation Multiple Buds cutting, be transferred to for the third time in proliferated culture medium, the formula of proliferated culture medium is for the third time: MS+6-BA1.5～2.5mg/L+NAA0.8～1.2mg/L, 24～26 DEG C of temperature, fluorescent lamp irradiate 24 hours/day, intensity of illumination 800～1200lx, cultivate 30～40 days, grow in incision the third generation Multiple Buds that 1cm is high, the propagation multiple of sprouting reaches more than 5 times, and starts to have the growth of adventive root;
7. strong seedling culture: will transfer in strong seedling culture base after the cutting of third generation Multiple Buds, this culture medium prescription is: MS+NAA1.0～1.5mg/L, condition of culture: 24～26 DEG C of temperature, fluorescent lamp irradiate 16 hours/day, intensity of illumination 2000～2400lx, cultivate 30～45 days, form the high indefinite bud of 2～3cm;
8. culture of rootage: indefinite bud is inoculated in root media, this culture medium prescription is: MS+NAA0.8～1.2mg/L, condition of culture: 24～26 DEG C of temperature, fluorescent lamp irradiate 16 hours/day, intensity of illumination 2000～2400lx, cultivate 60～75 days, every young plant forms 2～3 roots, long 2～the 4cm of root, diameter 1mm;
9. light training hardening: the seedling of having taken root is proceeded to booth together with blake bottle, is at 25～30 DEG C in temperature, places 7 days, removes bottle stopper, hardening 5～7 days, can take out seedling to transplant.
Above-described MS medium is to be to use at present the most general medium, has higher inorganic salt concentration, can ensure the mineral nutrition that tissue growth is required, can also accelerate the growth of callus.BA is: 6-benzyladenine is a kind of basic element of cell division.NAA is: α-naphthaleneacetic acid is a kind of auximone.
Taking the tender stem apex of children, axillalry bud or young stem stem section as explant material, because plant cell enlivens, add the plant growth regulator that concentration is lower to break up by rapid induction, but the dendrobium candidum stem section of the state of growing up, cell division speed slows down, this stem section is organized training as explant, and the axillary bud sprouting rate of younger tender stem apex, axillalry bud or young stem stem section is low, plant growth regulator add bad control.In test, find, the formula of proliferated culture medium is: when MS+6-BA4.8～5.2mg/L+NAA0.8～1.2mg/L, callus is too much, and reproduction speed is fast, but reaches more than 6 months when strong sprout, and output is crossed weak meticulous obsolete seedling; The formula of proliferated culture medium is: when MS+6-BA1.5～2.5mg/L+NAA0.8～1.2mg/L, result propagation is too slow, and each cycle 2～3 seedlings, do not reach fast numerous object; And taking proliferated culture medium: MS+6-BA3.8-4.2mg/L+NAA0.8～1.2mg/L as buffering, and in conjunction with the first two proliferation culture medium formula, can make the propagation multiple of sprouting reach 10000 times, form the healthy and strong Multiple Buds that about 1cm is high, shorten cultivation period, and can go out qualified dendrobium candidum training seedling by quickly breeding, for preparing strong sprout.
The invention has the beneficial effects as follows:
1. the present invention, taking the dendrobium candidum in more than 1 year vegetative period from the above stem section of root three joints as explant material carries out tissue-culturing rapid propagation, has advantages of and draws materials easy, quantity of material greatly, be not subject to seasonal restrictions.
2. the present invention adopts the stem section training method with blastogenesis bud, induction time section, and generating process is simple, does not have the forfeiture of tissue or the basic plant structure of differentiation, can keep preferably maternal biological character, thereby ensure the medical value of dendrobium candidum.
3. culture technique is simple, the sterilization method pollution rate adopting is low, reproduction speed fast (propagation multiple can reach 10000 times), cycle short (7～8 months), can increase reproductive speed and output, reduce group training production cost, significant to resources of medicinal plant regeneration in short supply in imminent danger and sustainable utilization.
Embodiment
Below in conjunction with embodiment, the invention will be further described, and so that its beneficial effect to be described, but the present invention is limited to absolutely not these examples.
Embodiment 1
Dendrobium candidum stem segment tissue culture fast breeding method, comprises the following steps:
1. the preparation of stem explants: the dendrobium candidum of getting 1 year vegetative period, from stem sections more than root three joints, is removed blade, rinses well with flowing water;
2. the sterilization of stem explants: the stem explants cleaning up is taken out, put in beaker, mercuric chloride solution with 0.1% soaks 5 minutes, with aseptic water washing 5 times, put on superclean bench, to be cut into length be 1～2cm, little stem section with 2～3 joints, then soak 3 minutes with 0.1% mercuric chloride solution, with aseptic water washing 5 times, for subsequent use;
3. induction is cultivated: the little stem section disinfecting is inoculated in aseptic inducing culture, this culture medium prescription is: MS+6-BA0.8mg/L+NAA0.8mg/L, condition of culture: 24～26 DEG C of temperature, fluorescent lamp irradiate 24 hours/day, intensity of illumination 800～1200lx, cultivate 40 days, stem section goes out internode director the sprouting that about 0.5cm is high;
4. propagation is cultivated for the first time: sprouting is transferred to for the first time in proliferated culture medium, the formula of proliferated culture medium is for the first time: MS+6-BA4.8mg/L+NAA0.8mg/L, 24～26 DEG C of temperature, fluorescent lamp irradiate 24 hours/day, intensity of illumination 800～1200lx, cultivate 30 days, grow in incision the first generation Multiple Buds that about 0.5cm is high, the propagation multiple of sprouting reaches 20 times;
5. propagation is cultivated for the second time: will after first generation Multiple Buds cutting, be transferred to for the second time in proliferated culture medium, the formula of proliferated culture medium is for the second time: MS+6-BA3.8mg/L+NAA0.8mg/L, 24～26 DEG C of temperature, fluorescent lamp irradiate 24 hours/day, intensity of illumination 800～1200lx, cultivate 30 days, grow in incision the second generation Multiple Buds that about 0.5cm is high, the propagation multiple of sprouting reaches 10 times;
6. propagation is cultivated for the third time: will after second generation Multiple Buds cutting, be transferred to for the third time in proliferated culture medium, the formula of proliferated culture medium is for the third time: MS+6-BA1.5mg/L+NAA0.8mg/L, 24～26 DEG C of temperature, fluorescent lamp irradiate 24 hours/day, intensity of illumination 800～1200lx, cultivate 40 days, grow in incision the third generation Multiple Buds that 1cm is high, the propagation multiple of sprouting reaches 5 times, and starts to have the growth of adventive root;
7. strong seedling culture: will transfer in strong seedling culture base after the cutting of third generation Multiple Buds, this culture medium prescription is: MS+NAA1mg/L medium, condition of culture: 24～26 DEG C of temperature, fluorescent lamp irradiate 16 hours/day, intensity of illumination 2000～2400lx, cultivate 30 days, form the high indefinite bud of 2～3cm;
8. culture of rootage: indefinite bud is inoculated in root media, this culture medium prescription is: MS+NAA0.8mg/L, condition of culture: 24～26 DEG C of temperature, fluorescent lamp irradiate 16 hours/day, intensity of illumination 2000～2400lx, cultivate 60 days, every young plant forms 2～3 roots, long 2～the 4cm of root, diameter 1mm;
9. light training hardening: the seedling of having taken root is proceeded to booth together with blake bottle, is at 25～30 DEG C in temperature, places 7 days, removes bottle stopper, hardening 7 days, can take out seedling to transplant.
Embodiment 2
Dendrobium candidum stem segment tissue culture fast breeding method, comprises the following steps:
1. the preparation of stem explants: the dendrobium candidum of getting 2 year vegetative period, from stem sections more than root three joints, is removed blade, rinses well with flowing water;
2. the sterilization of stem explants: the stem explants cleaning up is taken out, put in beaker, mercuric chloride solution with 0.1% soaks 5 minutes, with aseptic water washing 4 times, put on superclean bench, to be cut into length be 1～2cm, little stem section with 2～3 joints, then soak 3 minutes with 0.1% mercuric chloride solution, with aseptic water washing 4 times, for subsequent use;
3. induction is cultivated: the little stem section disinfecting is inoculated in aseptic inducing culture, this culture medium prescription is: MS+6-BA1.0mg/L+NAA1.0mg/L, condition of culture: 24～26 DEG C of temperature, fluorescent lamp irradiate 24 hours/day, intensity of illumination 800～1200lx, cultivate 40 days, stem section goes out internode director the sprouting that about 0.5cm is high;
4. propagation is cultivated for the first time: sprouting is transferred to for the first time in proliferated culture medium, the formula of proliferated culture medium is for the first time: MS+6-BA5.0mg/L+NAA1.0mg/L, 24～26 DEG C of temperature, fluorescent lamp irradiate 24 hours/day, intensity of illumination 800～1200lx, cultivate 30 days, grow in incision the first generation Multiple Buds that about 0.5cm is high, the propagation multiple of sprouting reaches 100 times;
5. propagation is cultivated for the second time: will after first generation Multiple Buds cutting, be transferred to for the second time in proliferated culture medium, the formula of proliferated culture medium is for the second time: MS+6-BA4.0mg/L+NAA1.0mg/L, 24～26 DEG C of temperature, fluorescent lamp irradiate 24 hours/day, intensity of illumination 800～1200lx, cultivate 35 days, grow in incision the second generation Multiple Buds that about 0.5cm is high, the propagation multiple of sprouting reaches 10 times;
6. propagation is cultivated for the third time: will after second generation Multiple Buds cutting, be transferred to for the third time in proliferated culture medium, the formula of proliferated culture medium is for the third time: MS+6-BA1.8mg/L+NAA1.0mg/L, 24～26 DEG C of temperature, fluorescent lamp irradiate 24 hours/day, intensity of illumination 800～1200lx, cultivate 30 days, grow in incision the third generation Multiple Buds that 1cm is high, the propagation multiple of sprouting reaches 10 times, and starts to have the growth of adventive root;
7. strong seedling culture: will transfer in strong seedling culture base after the cutting of third generation Multiple Buds, this culture medium prescription is: MS+NAA1.2mg/L medium, condition of culture: 24～26 DEG C of temperature, fluorescent lamp irradiate 16 hours/day, intensity of illumination 2000～2400lx, cultivate 45 days, form the high indefinite bud of 2～3cm;
8. culture of rootage: indefinite bud is inoculated in root media, this culture medium prescription is: MS+NAA1.0mg/L, condition of culture: 24～26 DEG C of temperature, fluorescent lamp irradiate 16 hours/day, intensity of illumination 2000～2400lx, cultivate 75 days, every young plant forms 2～3 roots, long 2～the 4cm of root, diameter 1mm;
9. light training hardening: the seedling of having taken root is proceeded to booth together with blake bottle, is at 25～30 DEG C in temperature, places 7 days, removes bottle stopper, hardening 5 days, can take out seedling to transplant.
Embodiment 3
Dendrobium candidum stem segment tissue culture fast breeding method, comprises the following steps:
1. the preparation of stem explants: the dendrobium candidum of getting 3 year vegetative period, from stem sections more than root three joints, is removed blade, rinses well with flowing water;
2. the sterilization of stem explants: the stem explants cleaning up is taken out, put in beaker, mercuric chloride solution with 0.1% soaks 5 minutes, with aseptic water washing 4 times, put on superclean bench, to be cut into length be 1～2cm, little stem section with 2～3 joints, then soak 3 minutes with 0.1% mercuric chloride solution, with aseptic water washing 5 times, for subsequent use;
3. induction is cultivated: the little stem section disinfecting is inoculated in aseptic inducing culture, this culture medium prescription is: MS+6-BA1.2mg/L+NAA1.2mg/L, condition of culture: 24～26 DEG C of temperature, fluorescent lamp irradiate 24 hours/day, intensity of illumination 800～1200lx, cultivate 40 days, stem section goes out internode director the sprouting that about 0.5cm is high;
4. propagation is cultivated for the first time: sprouting is transferred to for the first time in proliferated culture medium, the formula of proliferated culture medium is for the first time: MS+6-BA5.2mg/L+NAA1.2mg/L, 24～26 DEG C of temperature, fluorescent lamp irradiate 24 hours/day, intensity of illumination 800～1200lx, cultivate 30 days, grow in incision the first generation Multiple Buds that about 0.5cm is high, the propagation multiple of sprouting reaches 50 times;
5. propagation is cultivated for the second time: will after first generation Multiple Buds cutting, be transferred to for the second time in proliferated culture medium, the formula of proliferated culture medium is for the second time: MS+6-BA4.2mg/L+NAA1.2mg/L, 24～26 DEG C of temperature, fluorescent lamp irradiate 24 hours/day, intensity of illumination 800～1200lx, cultivate 40 days, grow in incision the second generation Multiple Buds that about 0.5cm is high, the propagation multiple of sprouting reaches 15 times;
6. propagation is cultivated for the third time: will after second generation Multiple Buds cutting, be transferred to for the third time in proliferated culture medium, the formula of proliferated culture medium is for the third time: MS+6-BA1.5mg/L+NAA0.8mg/L, 24～26 DEG C of temperature, fluorescent lamp irradiate 24 hours/day, intensity of illumination 800～1200lx, cultivate 35 days, grow in incision the third generation Multiple Buds that 1cm is high, the propagation multiple of sprouting reaches 8 times, and starts to have the growth of adventive root;
7. strong seedling culture: will transfer in strong seedling culture base after the cutting of third generation Multiple Buds, this culture medium prescription is: MS+NAA1.5mg/L medium, condition of culture: 24～26 DEG C of temperature, fluorescent lamp irradiate 16 hours/day, intensity of illumination 2000～2400lx, cultivate 40 days, form the high indefinite bud of 2～3cm;
8. culture of rootage: indefinite bud is inoculated in root media, this culture medium prescription is: MS+NAA0.8mg/L, condition of culture: 24～26 DEG C of temperature, fluorescent lamp irradiate 16 hours/day, intensity of illumination 2000～2400lx, cultivate 60 days, every young plant forms 2～3 roots, long 2～the 4cm of root, diameter 1mm;
9. light training hardening: the seedling of having taken root is proceeded to booth together with blake bottle, is at 25～30 DEG C in temperature, places 7 days, removes bottle stopper, hardening 6 days, can take out seedling to transplant.
Claims (1)
1. a dendrobium candidum stem segment tissue culture fast breeding method, is characterized in that, comprises the following steps:
(1) preparation of stem explants: get the state dendrobium candidum stem section of growing up, remove blade, rinse well with flowing water;
The dendrobium candidum that described adult state dendrobium candidum stem section is more than 1 year vegetative period is from stem sections more than root three joints;
(2) sterilization of stem explants: the stem explants cleaning up is taken out, put in beaker, mercuric chloride solution with 0.1% soaks 5 minutes, with aseptic water washing 4～5 times, put on superclean bench, to be cut into length be 1～2cm, little stem section with 2～3 joints, then soak 3 minutes with 0.1% mercuric chloride solution, with aseptic water washing 4～5 times, for subsequent use;
(3) induction is cultivated: the little stem section disinfecting is inoculated in aseptic inducing culture, this culture medium prescription is: MS+6-BA0.8～1.2mg/L+NAA0.8～1.2mg/L, condition of culture: 24～26 DEG C of temperature, fluorescent lamp irradiate 24 hours/day, intensity of illumination 800～1200lx, cultivate after 40 days, stem section goes out sprouting internode director;
(4) propagation is cultivated for the first time: sprouting is transferred to for the first time in proliferated culture medium, the formula of proliferated culture medium is for the first time: MS+6-BA4.8～5.2mg/L+NAA0.8～1.2mg/L, 24～26 DEG C of temperature, fluorescent lamp irradiate 24 hours/day, intensity of illumination 800～1200lx, cultivate after 30 days, grow first generation Multiple Buds in incision;
(5) propagation is cultivated for the second time: will after first generation Multiple Buds cutting, be transferred to for the second time in proliferated culture medium, the formula of proliferated culture medium is for the second time: MS+6-BA3.8～4.2mg/L+NAA0.8～1.2mg/L, 24～26 DEG C of temperature, fluorescent lamp irradiate 24 hours/day, intensity of illumination 800～1200lx, cultivate 30～40 days, grow second generation Multiple Buds in incision;
(6) propagation is cultivated for the third time: will after second generation Multiple Buds cutting, be transferred to for the third time in proliferated culture medium, the formula of proliferated culture medium is for the third time: MS+6-BA1.5～2.5mg/L+NAA0.8～1.2mg/L, 24～26 DEG C of temperature, fluorescent lamp irradiate 24 hours/day, intensity of illumination 800～1200lx, cultivate 30～40 days, grow in incision the third generation Multiple Buds that 1cm is high, and start to have adventive root growth;
(7) strong seedling culture: will transfer in strong seedling culture base after the cutting of third generation Multiple Buds, this culture medium prescription is: MS+NAA1.0～1.5mg/L, condition of culture: 24～26 DEG C of temperature, fluorescent lamp irradiate 16 hours/day, intensity of illumination 2000～2400lx, cultivate 30～45 days, form the high indefinite bud of 2～3cm;
(8) culture of rootage: indefinite bud is inoculated in root media, this culture medium prescription is: MS+NAA0.8～1.2mg/L, condition of culture: 24～26 DEG C of temperature, fluorescent lamp irradiate 16 hours/day, intensity of illumination 2000～2400lx, cultivate 60～75 days, every young plant forms 2～3 roots, long 2～the 4cm of root, diameter 1mm;
(9) light training hardening: the seedling of having taken root is proceeded to booth together with blake bottle, is at 25～30 DEG C in temperature, places 7 days, removes bottle stopper, hardening 5～7 days, can take out seedling to transplant.
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| CN103609447A (en) * | 2013-11-27 | 2014-03-05 | 苏州田园农业技术开发有限公司 | Propagation method of Dendrobium officinale |
| CN104429961B (en) * | 2014-12-08 | 2016-08-24 | 河源市裕森农林发展有限公司 | A kind of Fast-propagation tissue culture method of Herba Dendrobii |
| CN106258981A (en) * | 2016-08-31 | 2017-01-04 | 耿马四方生物科技开发有限责任公司 | Candidum tissue culturing factorial seedling-culturing method |
| CN107347649A (en) * | 2017-08-11 | 2017-11-17 | 佛山科学技术学院 | A kind of cultural method of dendrobium candidum explant |
| CN109984035A (en) * | 2019-03-22 | 2019-07-09 | 扬州雅典娜园艺科技开发有限公司 | A method of establishing HERBA DENDROBII regenerating system |
| CN115176608B (en) * | 2022-06-30 | 2024-02-13 | 贵州省林业科学研究院 | Rapid seedling method for dendrobium candidum stem node induced buds |
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| CN101258835B (en) * | 2008-04-23 | 2010-06-16 | 昆明理工大学 | Fast reproducing method for high quality seedling of dendrobium officinale |
| CN102369881B (en) * | 2011-09-22 | 2014-05-07 | 澄思源生物科技（上海）有限公司 | Rapid propagation technique of Dendrobium candidum axillary buds |
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