CN104255457A - Rapid propagation method for tissue culture of ilex nitidissima - Google Patents
Rapid propagation method for tissue culture of ilex nitidissima Download PDFInfo
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- CN104255457A CN104255457A CN201410462993.4A CN201410462993A CN104255457A CN 104255457 A CN104255457 A CN 104255457A CN 201410462993 A CN201410462993 A CN 201410462993A CN 104255457 A CN104255457 A CN 104255457A
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Abstract
The invention discloses a rapid propagation method for tissue culture of ilex nitidissima. The method comprises the steps of obtaining sterile terminal buds, inducing cluster buds, carrying out propagation on the cluster buds, carrying out rooting culture, exercising seedlings, etc. The tissue culture technical method can be used for carrying out tissue culture micro-propagation on the ilex nitidissima, so that the technical problems that the ilex nitidissima is high in contamination rate, low in breeding rate and the like can be solved, and the requirements on high multiplication coefficient and high survival rate can be met; therefore, an artificial cultivation method with short period can be provided for the industrialized production of the ilex nitidissima.
Description
Technical field
The present invention relates to a kind of tissue culture quick propagation culturing method of Ilex viridis Champ, belong to biological technical field.
Background technology
Ilex viridis Champ,
ilex nitidissima C. J. Tseng, Aquifoliaceae, evergreen dungarunga, originates in Jiangxi, China northeast, Southern Hunan and northeast of Guangxi and south, Gao Keda 6 meters; Blade keratin, back side tool gland point or nothing, tip shape of tail is gradually sharp; Inflorescence clusters, each branch tool 3 flower of male inflorescence, each branch tool 1 flower of female inflorescence, rare tool 2 ~ 5 flower; Spend 4 radixes, rare 5 or 6 radixes; Spherical or the cushion of abortion ovary, the short beak of tool; Fruit is spherical, diameter about 5 millimeters, time ripe, red, the striped of tool protuberance, endocarp keratin.Really 7 ~ October of phase.Be born in the hillside thick forest of height above sea level 960 ~ 1250 meters, sparse woods or shaw.For the root leaf of holly plant Ilex viridis Champ.Bark is resinous.Effect: removing pattogenic heat from the blood and toxic material from the body, to remove obstruction in channels to relieve pain.Cure mainly: control hot carbuncle toxin, dysentery, fall wink frustrate, hemostasis is had a pain, beriberoid pyretic arthralgia.Meridian distribution of property and flavor: bitter, micro-pungent, cool, enters the heart, liver two warp, and the main routine that adopts is planted at present, and tissue culture method can obtain a large amount of Ilex viridis Champ seedling fast, keeps the stability of genetic character, overcomes the limitation of region.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of tissue culture quick propagation culturing method of Ilex viridis Champ, growth coefficient is reached good by the preparation-obtained Ilex viridis Champ of the method, the requirement that survival rate is high, can produce for its industrialization the artificial culturing method providing a kind of cycle short.
Technical problem to be solved by this invention is realized by following scheme:
Get Ilex viridis Champ terminal bud, cut off unnecessary blade, retain a leaflet, remove dirt with hairbrush, add bleaching powder and soak 3 minutes, running water 0.5 hour, use 10% antiformin solution-treated 6min, aseptic water washing 5-7 time, cuts otch, and the aseptic bud grafting kind of acquisition enters B
5+ NAA0.1mg/L+KT0.5mg/L+2,4-D0.1mg/L+300 μ g/g benzyl ammonia carries out inducing clumping bud, and condition of culture adopts the lower cultivation by force of the 800lx low light level, sucrose 30g/L, agar 6.5g/L, pH value 5.8, temperature 25 DEG C, photophase 8h, dark phase 16h, the Ilex viridis Champ Multiple Buds derived is transferred in B
5+ 6-BA1.5mg/L+IBA0.1mg/L carries out adventitious buds proliferation cultivation, and condition of culture is illumination 2000lx, sucrose 30g/L, agar 6.5g/L, pH value 5.8, temperature 27 DEG C, photophase 12h, dark phase 12h, and after the Multiple Buds process of propagation, access is containing 1/2 B
5carry out root induction in the medium of+IBA0.05mg/L+AC1.0g/L, condition of culture illumination 6000lx, photophase 16h, dark phase 8h, the test-tube plantlet being about 5cm takes out plants in count's mud stone 525
#: in the matrix of vermiculite=3:2, spray foliage fertilizer after three weeks, humidity 85%, temperature 25 DEG C, hide double-layer films, light transmittance remains on about 35%.
The Ilex viridis Champ group training seedling adopting the present invention to prepare is with short production cycle, reproduction rate is high, rooting rate 92%.
Below in conjunction with embodiment, the present invention is further elaborated, but the scope of protection of present invention is not limited to following embodiments.
Embodiment
Embodiment 1
Get Ilex viridis Champ terminal bud, cut off unnecessary blade, retain a leaflet, remove dirt with hairbrush, add bleaching powder and soak 3 minutes, running water 0.5 hour, use 10% antiformin solution-treated 6min, aseptic water washing 5-7 time, cuts otch, and the aseptic bud grafting kind of acquisition enters B
5+ NAA0.1mg/L+KT0.5mg/L+2,4-D0.1mg/L+300 μ g/g benzyl ammonia carries out inducing clumping bud, and condition of culture adopts the lower cultivation by force of the 800lx low light level, sucrose 30g/L, agar 6.5g/L, pH value 5.8, temperature 25 DEG C, photophase 8h, dark phase 16h, the Ilex viridis Champ Multiple Buds derived is transferred in B
5+ 6-BA1.0mg/L+IBA0.05mg/L carries out adventitious buds proliferation cultivation, and condition of culture is illumination 2000lx, sucrose 30g/L, agar 6.5g/L, pH value 5.8, temperature 27 DEG C, photophase 12h, dark phase 12h, and after the Multiple Buds process of propagation, access is containing 1/2 B
5carry out root induction in the medium of+IBA0.05mg/L+AC0.5g/L, condition of culture illumination 6000lx, photophase 16h, dark phase 8h, the test-tube plantlet being about 5cm takes out plants in count's mud stone 525
#: in the matrix of vermiculite=3:2, spray foliage fertilizer after three weeks, humidity 85%, temperature 25 DEG C, hide double-layer films, light transmittance remains on about 35%, and rooting rate is 86%.
Embodiment 2
Get Ilex viridis Champ terminal bud, cut off unnecessary blade, retain a leaflet, remove dirt with hairbrush, add bleaching powder and soak 3 minutes, running water 0.5 hour, use 10% antiformin solution-treated 6min, aseptic water washing 5-7 time, cuts otch, and the aseptic bud grafting kind of acquisition enters B
5+ NAA0.1mg/L+KT0.5mg/L+2,4-D0.1mg/L+300 μ g/g benzyl ammonia carries out inducing clumping bud, and condition of culture adopts the lower cultivation by force of the 800lx low light level, sucrose 30g/L, agar 6.5g/L, pH value 5.8, temperature 25 DEG C, photophase 8h, dark phase 16h, the Ilex viridis Champ Multiple Buds derived is transferred in B
5+ 6-BA1.5mg/L+IBA0.1mg/L carries out adventitious buds proliferation cultivation, and condition of culture is illumination 2000lx, sucrose 30g/L, agar 6.5g/L, pH value 5.8, temperature 27 DEG C, photophase 12h, dark phase 12h, and after the Multiple Buds process of propagation, access is containing 1/2 B
5carry out root induction in the medium of+IBA0.05mg/L+AC1.5g/L, condition of culture illumination 6000lx, photophase 16h, dark phase 8h, the test-tube plantlet being about 5cm takes out plants in count's mud stone 525
#: in the matrix of vermiculite=3:2, spray foliage fertilizer after three weeks, humidity 85%, temperature 25 DEG C, hide double-layer films, light transmittance remains on about 35%, and rooting rate is 87%.
Embodiment 3
Get Ilex viridis Champ terminal bud, cut off unnecessary blade, retain a leaflet, remove dirt with hairbrush, add bleaching powder and soak 3 minutes, running water 0.5 hour, use 10% antiformin solution-treated 6min, aseptic water washing 5-7 time, cuts otch, and the aseptic bud grafting kind of acquisition enters B
5+ NAA0.1mg/L+KT0.5mg/L+2,4-D0.1mg/L+300 μ g/g benzyl ammonia carries out inducing clumping bud, and condition of culture adopts the lower cultivation by force of the 800lx low light level, sucrose 30g/L, agar 6.5g/L, pH value 5.8, temperature 25 DEG C, photophase 8h, dark phase 16h, the Ilex viridis Champ Multiple Buds derived is transferred in B
5+ 6-BA1.5mg/L+IBA0.05mg/L carries out adventitious buds proliferation cultivation, and condition of culture is illumination 2000lx, sucrose 30g/L, agar 6.5g/L, pH value 5.8, temperature 27 DEG C, photophase 12h, dark phase 12h, and after the Multiple Buds process of propagation, access is containing 1/2 B
5carry out root induction in the medium of+IBA0.05mg/L+AC1.0g/L, condition of culture illumination 6000lx, photophase 16h, dark phase 8h, the test-tube plantlet being about 5cm takes out plants in count's mud stone 525
#: in the matrix of vermiculite=3:2, spray foliage fertilizer after three weeks, humidity 85%, temperature 25 DEG C, hide double-layer films, light transmittance remains on about 35%, and rooting rate is 89%.
Claims (6)
1. a tissue culture quick propagation culturing method for Ilex viridis Champ, its step is as follows:
(1) acquisition of aseptic terminal bud: get the Ilex viridis Champ terminal bud that children is tender, to its disinfection;
(2) bud inducement is cultivated: aseptic bud grafting kind step (1) obtained enters B
5+ NAA0.1mg/L+KT0.5mg/L+2,4-D0.1mg/L+300 μ g/g benzyl ammonia carries out inducing clumping bud;
(3) adventitious buds proliferation is cultivated: the Ilex viridis Champ Multiple Buds derived in step (2) is transferred in B
5+ 6-BA1-1.5mg/L+IBA0.1mg/L carries out adventitious buds proliferation cultivation;
(4) root induction: after the Multiple Buds process of breed step (3), access is containing 1/2 B
5root induction is carried out in the medium of+IBA0.05-0.1mg/L+AC0.5-1.5g/L;
(5) hardening: the test-tube plantlet after taking root is planted on seedbed, adds up survival rate after month.
2. the tissue culture quick propagation culturing method of a kind of Ilex viridis Champ according to claim 1, it is characterized in that: the sterilization method of Ilex viridis Champ terminal bud is for getting Ilex viridis Champ terminal bud, cut off unnecessary blade, retain a leaflet, remove dirt with hairbrush, add bleaching powder and soak 3 minutes, running water 0.5 hour, use 10% antiformin solution-treated 6min, aseptic water washing 5-7 time, cuts otch.
3. the tissue culture quick propagation culturing method of a kind of Ilex viridis Champ according to claim 1, is characterized in that: add benzyl ammonia in medium, effectively can control the generation of bacterium.
4. the tissue culture quick propagation culturing method of a kind of Ilex viridis Champ according to claim 1, it is characterized in that: add adsorbent A C in root media, AC is conducive to hestening rooting, and can prevent the generation of browning.
5. the tissue culture quick propagation culturing method of a kind of Ilex viridis Champ according to claim 1, is characterized in that: inducing culturing condition adopts the lower cultivation by force of the 800lx low light level, sucrose 30g/L, agar 6.5g/L, pH value 5.8, temperature 25 DEG C, photophase 8h, dark phase 16h, Multiplying culture condition is illumination 2000lx, sucrose 30g/L, agar 6.5g/L, pH value 5.8, temperature 27 DEG C, photophase 12h, dark phase 12h, culture of rootage condition illumination 6000lx, photophase 16h, dark phase 8h.
6. the tissue culture quick propagation culturing method of a kind of Ilex viridis Champ according to claim 1, is characterized in that: the medium of seedling bed of hardening selects import count mud stone 525
#: vermiculite=3:2, sprayed foliage fertilizer after three weeks, humidity 85%, temperature 25 DEG C, and hide double-layer films, light transmittance remains on about 35%.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106258952A (en) * | 2015-12-02 | 2017-01-04 | 香港中文大学深圳研究院 | The method utilizing Caulis et Folium Ammopiptanthi Mongolici children tender side root induction Caulis et Folium Ammopiptanthi Mongolici Callus formation |
| CN107173231A (en) * | 2017-07-03 | 2017-09-19 | 甘肃省科学院生物研究所 | A kind of method of the Mongolian Ammopiptanthus mongolicus rapid propagation in vitro of rare or endangered species |
| CN108029559A (en) * | 2017-12-29 | 2018-05-15 | 南京林业大学 | A kind of method of quickly breeding bearberry tissue-cultured seedling |
| CN112493124A (en) * | 2020-11-09 | 2021-03-16 | 江苏农林职业技术学院 | Culture medium and culture method for promoting subculture bud differentiation of ilex chinensis |
| CN115067140A (en) * | 2022-07-20 | 2022-09-20 | 惠州市林业科学研究所(惠州植物园管理服务中心) | Rapid seedling raising method for ilex latifolia seeds |
| CN115250909A (en) * | 2022-06-14 | 2022-11-01 | 江苏青好景观园艺有限公司 | Cultivation method of Stefin holly tissue culture seedling |
-
2014
- 2014-09-12 CN CN201410462993.4A patent/CN104255457A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106258952A (en) * | 2015-12-02 | 2017-01-04 | 香港中文大学深圳研究院 | The method utilizing Caulis et Folium Ammopiptanthi Mongolici children tender side root induction Caulis et Folium Ammopiptanthi Mongolici Callus formation |
| CN106258952B (en) * | 2015-12-02 | 2018-11-09 | 香港中文大学深圳研究院 | The method formed using the tender lateral root induction Ammopiptanthus mongolicus callus of Ammopiptanthus mongolicus children |
| CN107173231A (en) * | 2017-07-03 | 2017-09-19 | 甘肃省科学院生物研究所 | A kind of method of the Mongolian Ammopiptanthus mongolicus rapid propagation in vitro of rare or endangered species |
| CN108029559A (en) * | 2017-12-29 | 2018-05-15 | 南京林业大学 | A kind of method of quickly breeding bearberry tissue-cultured seedling |
| CN112493124A (en) * | 2020-11-09 | 2021-03-16 | 江苏农林职业技术学院 | Culture medium and culture method for promoting subculture bud differentiation of ilex chinensis |
| CN115250909A (en) * | 2022-06-14 | 2022-11-01 | 江苏青好景观园艺有限公司 | Cultivation method of Stefin holly tissue culture seedling |
| CN115067140A (en) * | 2022-07-20 | 2022-09-20 | 惠州市林业科学研究所(惠州植物园管理服务中心) | Rapid seedling raising method for ilex latifolia seeds |
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Application publication date: 20150107 |