CN104920225A - Acer negundo 'Aurea' tissue culture rapid propagation method - Google Patents
Acer negundo 'Aurea' tissue culture rapid propagation method Download PDFInfo
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- CN104920225A CN104920225A CN201510411373.2A CN201510411373A CN104920225A CN 104920225 A CN104920225 A CN 104920225A CN 201510411373 A CN201510411373 A CN 201510411373A CN 104920225 A CN104920225 A CN 104920225A
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Abstract
The invention discloses an Acer negundo 'Aurea' tissue culture rapid propagation method. The method includes the following steps that newly grown twigs about 5 cm in length are collected after spring dormant buds sprouting, the twigs are inoculated into an MS culture medium into which 6- benzylaminopurine with the concentration ranging from 0.5 mg/L -1.0 mg/L is added after the surfaces of the twigs are sterilized, and axillary buds are induced to sprout; obtained tender stems are clipped into segments ranging from 2 cm to 3 cm in length and inoculated into the MS culture medium, and enrichment culture is conducted; when single buds growing strongly and ranging from 3 cm to 4 cm in length are inoculated into the 1/2 MS culture medium, rooting culture is conducted; when the roots grow to 3 cm to 6 cm, a bottle cover is opened, and seedlings are transplanted 3 days after hardening-seedling is conducted. According to the method, the detached material axillary buds of the Acer negundo 'Aurea' can be promoted to sprout quickly by adjusting the culture medium components and controlling the culture conditions, test pipe seedling stem segments are induced to directly generate adventitious buds without a callus, a great number of fibrous roots exist, the transplant survival rate reaches 85%, and an effective way is provided for industrialized seedling production of the Acer negundo 'Aurea'.
Description
Technical field
The present invention relates to method for plant tissue culture, be specifically related to a kind of tissue culture and rapid propagation method of gold leaf Acer negundo. L.
Background technology
Gold leaf Acer negundo. L (Acer negundo ' Aurea ') belongs to Aceraceae (Aceraceae) Acer (Acer) seeds, it is the cultivar of Acer negundo. L, deciduous tree is the excellent ornamental tree species that China introduced from Europe in recent years.Gold leaf Acer negundo. L growth potential is vigorous, and sprig is smooth, golden yellow; Imparipinnate leaf is to life, and leaf is comparatively large, and leaf look soft, and spring is golden yellow, fades to yellow green, has high ornamental value.Gold leaf Acer negundo. L Bioclimatic analysis is wide, resistance, and drought-resistant, cold resistant (can resistance to-40-45 DEG C of low temperature), resistance to slight alkaline land, resistance to flue dust, root sprout tillers is strong.The suitable natural disposition stronger because of it and higher ornamental value, Landscape Trees cultivation is all done in China North China, northeast, northwest and East China, and supply falls short of demand for existing market.
In production, the main modes of reproduction of gold leaf Acer negundo. L is propagation by grafiting.Propagation by grafiting survival rate is higher, but the improper later stage tree vigo(u)r that can affect of Root-stock selection grows, and reproduction coefficient is lower, input cost is larger.Tissue culture technique is one of numerous soon main path of breeding, and reproduction coefficient is high, makes a variation little.Report about gold leaf Acer negundo. L tissue cultures is little, and just to the pre-test of its group training performance, the tissue cultures with other Aceraceae seeds is the same, there is sprouting and starts the defect slow, growth coefficient is low, poor-performing of taking root, survival rate are low.
Summary of the invention
The invention provides a kind of tissue culture and rapid propagation method of gold leaf Acer negundo. L, overcome in the group training cultivation of gold leaf Acer negundo. L and start the deficiency slow, growth coefficient is low, poor performance of taking root, survival rate are low.
For achieving the above object, the technical scheme that the present invention takes is:
A tissue culture and rapid propagation method for gold leaf Acer negundo. L, comprises the steps:
S1, after sleeping bud stripping, gather the newborn spray of about 5cm spring, be inoculated in the MS medium of 6-benzylaminopurine (6-BA) of additional 0.5-1.0mg/L after surface sterilization, in 20 ± 2 DEG C-25 ± 2 DEG C, 14 h light, cultivate under intensity of illumination 2000lx condition, induction axillary bud sprouting is to the tender stem of 6-8cm length;
S2, get step S1 obtain tender stem, be cut into the sections of 2-3cm with 2 petioles, be inoculated in the MS medium of additional 0.5mg/L 6-benzylaminopurine (6-BA), 0.01-0.05mg/L Thidiazuron (TDZ), 0.05mg/L methyl α-naphthyl acetate (NAA), in 20 ± 2 DEG C-25 ± 2 DEG C, 14 h light, under intensity of illumination 2000lx condition, carry out Multiplying culture;
S3, the robust growth of selecting step S2 gained, the simple bud of long 3-4cm are inoculated in the 1/2MS medium containing 0.01-0.05mg/L indolebutyric acid (IBA), in 20 ± 2 DEG C-25 ± 2 DEG C, 14 h light, under intensity of illumination 2000lx condition, carry out culture of rootage.
S4, when root grows to 3-6cm, bottle cap is opened, hardening is after 3 days, seedling is pressed from both sides out gently with tweezers, in flowing water, rinse root medium gently, transplant in the disposal plastic cup that matrix is housed, be placed in plastics valve bag by every for plastic cup 10, room temperature hardening, after 30 days, grows potted plant on seedling of taking root in the matrix containing perlite, vermiculite and rural area soil.
Wherein, 6g/L agar powder, 30g/L sucrose is also comprised in the medium in described step S1.
Wherein, in described step S2, proliferated culture medium also comprises 6g/L agar powder, 30g/L sucrose, 0.25g/L MES (MES), 0.2g/L l-glutamine.
Wherein, in described step S3, root media also comprises 7g/L agar powder, 15g/L sucrose, 0.25g/L MES (MES).
Wherein, in described step S4 disposal plastic cup matrix by perlite and peat soil in 1: 1 ratio mix.
Wherein, in described step S4, the ratio of perlite, vermiculite, rural area soil is 3: 2: 5.
The present invention has following beneficial effect:
Solve sprouting in Aceraceae groups of tree species training process and start the problem slow, growth coefficient is low, taking root property difference, transplanting survival rate are low.By the adjustment of nutrient media components and the control of condition of culture, comparatively fast can start gold leaf Acer negundo. L To body material axillary bud sprouting, induction test-tube plantlet stem section directly produces the more indefinite bud of number without callus, average each explant propagation bud subnumber can reach 5.8, adventitious bud rooting rate reaches 100%, and main root is flourishing, and fibrous root quantity is many, transplanting survival rate reaches 85%, for gold leaf Acer negundo. L factorial seedling growth provides an effective way.
Embodiment
In order to make objects and advantages of the present invention clearly understand, below in conjunction with embodiment, the present invention is further elaborated.Should be appreciated that specific embodiment described herein only in order to explain the present invention, be not intended to limit the present invention.
Embodiment 1
The newborn spray of about 5cm is gathered after sleeping bud stripping spring, be inoculated in the MS medium containing the 6-BA (0,0.5,1.0mg/L) of variable concentrations gradient, 6g/L agar powder, 30g/L sucrose after surface sterilization, in 25 ± 2 DEG C, 14 h light, cultivate under intensity of illumination 2000lx condition, induction axillary bud sprouting.Statistics axillary bud sprouting time and sprouting stem section state.Result shows, 25 days, 15 days, 12 days are respectively containing the axillary bud sprouting time of explant in the medium of 0mg/L, 0.5mg/L, 1.0mg/L 6-BA, add 6-BA and can shift to an earlier date axillalry bud start-up time, normal containing the tender bulbous state sprouted in 0.5-1.0mg/L 6-BA medium, blade is open and flat, sprouts tender stem and extends speed.
Embodiment 2
The newborn spray of about 5cm is gathered after sleeping bud stripping spring, be inoculated in the MS medium containing 0.5mg/L 6-BA, 6g/L agar powder, 30g/L sucrose after surface sterilization, in 25 ± 2 DEG C, 14 h light, cultivate under intensity of illumination 2000lx condition, induction axillary bud sprouting.By the tender stem that axillary bud sprouting obtains, be cut into the sections of 2-3cm with 2 petioles, be seeded in containing 0.5mg/L 6BA, 0.05mg/L NAA, variable concentrations gradient TDZ (0,0.01,0.05,0.1mg/L), 6g/L agar powder, 30g/L sucrose, 0.25g/L MES, 0.2g/L l-glutamine MS medium in, in 25 ± 2 DEG C, 14 h light, cultivate under intensity of illumination 2000lx condition and carry out Multiplying culture.Cultivate and add up proliferation times after 50 days.Result shows, in the medium containing 0mg/L, 0.01mg/L, 0.05mg/L, 0.1mg/L TDZ, the bud subnumber of average each stem section propagation is respectively 2,4.8,5.8,2.6.First three bud growth of breeding under planting concentration is normal, and the bud tubbiness of breeding under rear a kind of concentration, distortion, explant incision callusization is serious.
Embodiment 3
The newborn spray of about 5cm is gathered after sleeping bud stripping spring, be inoculated in the MS medium containing 0.5mg/L 6-BA, 6g/L agar powder, 30g/L sucrose after surface sterilization, in 25 ± 2 DEG C, 14 h light, cultivate under intensity of illumination 2000lx condition, induction axillary bud sprouting.By the tender stem that axillary bud sprouting obtains, be cut into the sections of 2-3cm with 2 petioles, be seeded in the MS medium containing 0.5mg/L 6BA, 0.05mg/L NAA, 0.05mg/L TDZ, 6g/L agar powder, 30g/L sucrose, 0.25g/L MES, 0.2g/L l-glutamine, in 25 ± 2 DEG C, 14 h light, cultivate under intensity of illumination 2000lx condition and carry out Multiplying culture.Robust growth propagation obtained, the simple bud of long 3-4cm are seeded in the IBA of 1/2MS annex variable concentrations gradient (0.01,0.05,0.1mg/L) medium respectively, and carry out culture of rootage in the NAA of 1/2MS annex variable concentrations gradient (0.01,0.05,0.1mg/L) medium, also comprise 7g/L agar powder, 15g/L sucrose, 0.25g/L MES in various medium.Condition of culture 25 ± 2 DEG C, 14 h light, intensity of illumination 2000lx.Statistics rootage duration, the rooting rate after 50 days, observed and recorded are taken root situation.Result shows, the adventitious bud rooting cultivated is slow in containing the root media of NAA, root is short, thin and delicate, and main root is not obvious, and rootage duration is 21 days the earliest, maximum rooting rate 62%, the long 1.54cm of average root.Containing 0.01,0.05, the indefinite bud cultivated in 0.1mg/L IBA root media the earliest rootage duration be respectively 15,11,11 days, rooting rate all reaches 100%, and average root is long is respectively 4.2cm, 4.8cm, 5.1cm; The indefinite bud root growth cultivated in medium containing 0.01-0.05mg/L IBA is fast, root is sturdy, and main root is obvious, fibrous root is many, occurs callus lines containing the Gen Yujing junction, indefinite bud lower end of cultivating in 0.1mg/L IBA medium.
Embodiment 4
The newborn spray of about 5cm is gathered after sleeping bud stripping spring, be inoculated in the MS medium containing 0.5mg/L 6-BA, 6g/L agar powder, 30g/L sucrose after surface sterilization, in 25 ± 2 DEG C, 14 h light, cultivate under intensity of illumination 2000lx condition, induction axillary bud sprouting.By the tender stem that axillary bud sprouting obtains, be cut into the sections of 2-3cm with 2 petioles, be seeded in the MS medium containing 0.5mg/L 6BA, 0.05mg/L NAA, 0.05mg/L TDZ, 6g/L agar powder, 30g/L sucrose, 0.25g/L MES, 0.2g/L l-glutamine, in 25 ± 2 DEG C, 14 h light, cultivate under intensity of illumination 2000l x condition and carry out Multiplying culture.Robust growth propagation obtained, the simple bud of long 3-4cm are seeded in the 1/2MS medium containing 0.05mg/L IBA, 7g/L agar powder, 15g/L sucrose, 0.25g/L MES, in 25 ± 2 DEG C, and 14 h light, culture of rootage under intensity of illumination 2000lx condition.When root grows to 3-6cm, bottle cap is opened, hardening 3 days.First press from both sides out seedling gently with tweezers, root medium is rinsed gently in flowing water, transplant to matrix (perlite: peat soil=1: in disposal plastic cup 1) is housed, plastics valve bag is placed in by every for plastic cup 10, room temperature hardening is after 30 days, investigation survival rate, grows potted plant on seedling of taking root in the matrix of perlite+vermiculite+rural area soil (ratio 3: 2: 5).Result shows, carries out the tissue-culturing rapid propagation of gold leaf Acer negundo. L by this program, and after the explant inoculation that field gathers, axillalry bud starts and can start after 15 days in cultivation; Start the sterilizable material proliferation times average out to 5.8 obtained, reach as high as 6.2; Adventitious bud rooting rate reaches 100%, and the sturdy prosperity of root system, transplanting survival rate reaches 78%.
Embodiment 5
The newborn spray of about 5cm is gathered after sleeping bud stripping spring, be inoculated in the MS medium containing 0.5mg/L 6-BA, 6g/L agar powder, 30g/L sucrose after surface sterilization, in 20 ± 2 DEG C, 14 h light, cultivate under intensity of illumination 2000l x condition, induction axillary bud sprouting.By the tender stem that axillary bud sprouting obtains, be cut into the sections of 2-3cm with 2 petioles, be seeded in the MS medium containing 0.5mg/L 6BA, 0.05mg/L NAA, 0.05mg/L TDZ, 6g/L agar powder, 30g/L sucrose, 0.25g/L MES, 0.2g/L l-glutamine, in 20 ± 2 DEG C, 14 h light, cultivate under intensity of illumination 2000lx condition and carry out Multiplying culture.Robust growth propagation obtained, the simple bud of long 3-4cm are seeded in the 1/2MS medium containing 0.05mg/L IBA, 7g/L agar powder, 15g/L sucrose, 0.25g/L MES, in 20 ± 2 DEG C, and 14 h light, culture of rootage under intensity of illumination 2000l x condition.When root grows to 3-6cm, bottle cap is opened, hardening 3 days.First press from both sides out seedling gently with tweezers, root medium is rinsed gently in flowing water, transplant to matrix (perlite: peat soil=1: in disposal plastic cup 1) is housed, plastics valve bag is placed in by every for plastic cup 10, room temperature hardening is after 30 days, investigation survival rate, grows potted plant on seedling of taking root in the matrix of perlite+vermiculite+rural area soil (ratio 3: 2: 5).The present embodiment difference unique with embodiment 4 is that cultivation temperature is different.Result shows, the test-tube plantlet cultivated under 25 ± 2 DEG C of conditions, and the phase is as replaced medium not in time after incubation, and lower plate blade there will be slight chlorosis albinism, affects transplanting survival rate; After cultivation temperature is down to 20 ± 2 DEG C, effectively can stop test-tube plantlet lower blade chlorosis albinism, improve transplanting survival rate, the transplanting survival rate at this temperature is 85%.Two kinds of temperature are not remarkable in bud start-up time, proliferation times, aspect of performance of taking root impact.
In sum, by regulating the ratio of hormone in medium, control condition of culture, significantly can shift to an earlier date the bud start-up time of gold leaf Acer negundo. L tissue cultures, improve test-tube plantlet growth coefficient, increase performance of taking root, promote transplanting survival rate, the average each explant propagation bud subnumber of cultivation method of the present invention can reach 5.8, adventitious bud rooting rate reaches 100%, and main root is flourishing, and fibrous root quantity is many, transplanting survival rate reaches 85%, for gold leaf Acer negundo. L factorial seedling growth provides an effective way.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (6)
1. a tissue culture and rapid propagation method for gold leaf Acer negundo. L, is characterized in that, comprises the steps:
S1, after sleeping bud stripping, gather the newborn spray of about 5cm spring, be inoculated in the MS medium of the 6-benzylaminopurine of additional 0.5-1.0mg/L after surface sterilization, in 20 ± 2 DEG C-25 ± 2 DEG C, 14 h light, cultivate under intensity of illumination 2000lx condition, induction axillary bud sprouting is to the tender stem of 6-8cm length;
S2, get step S1 obtain tender stem, be cut into the sections of 2-3cm with 2 petioles, be inoculated in the MS medium of additional 0.5mg/L 6-benzylaminopurine, 0.01-0.05mg/L Thidiazuron, 0.05mg/L methyl α-naphthyl acetate, in 20 ± 2 DEG C-25 ± 2 DEG C, 14 h light, under intensity of illumination 2000lx condition, carry out Multiplying culture;
S3, the robust growth of selecting step S2 gained, the simple bud of long 3-4cm are inoculated in the 1/2MS medium containing 0.01-0.05mg/L indolebutyric acid, and in 20 ± 2 DEG C-25 ± 2 DEG C, 14 h light, under intensity of illumination 2000lx condition, carry out culture of rootage.
S4, when root grows to 3-6cm, bottle cap is opened, hardening is after 3 days, seedling is pressed from both sides out gently with tweezers, in flowing water, rinse root medium gently, transplant in the disposal plastic cup that matrix is housed, be placed in plastics valve bag by every for plastic cup 10, room temperature hardening, after 30 days, grows potted plant on seedling of taking root in the matrix containing perlite, vermiculite and rural area soil.
2. gold leaf Acer negundo. L tissue culture and rapid propagation method according to claim 1, is characterized in that, also comprises 6g/L agar powder, 30g/L sucrose in the medium in described step S1.
3. gold leaf Acer negundo. L tissue culture and rapid propagation method according to claim 1, is characterized in that, in described step S2, proliferated culture medium also comprises 6g/L agar powder, 30g/L sucrose, 0.25g/L MES, 0.2g/L l-glutamine.
4. gold leaf Acer negundo. L tissue culture and rapid propagation method according to claim 1, is characterized in that, in described step S3, root media also comprises 7g/L agar powder, 15g/L sucrose, 0.25g/L MES.
5. gold leaf Acer negundo. L tissue culture and rapid propagation method according to claim 1, is characterized in that, in described step S4 disposal plastic cup matrix by perlite and peat soil in 1: 1 ratio mix.
6. gold leaf Acer negundo. L tissue culture and rapid propagation method according to claim 1, is characterized in that, in described step S4, the ratio of perlite, vermiculite, rural area soil is 3: 2: 5.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106305427A (en) * | 2016-08-30 | 2017-01-11 | 长江三峡生态园林有限公司 | Tissue culture and rapid propagation method for Acer ginnala |
| CN107455261A (en) * | 2017-09-15 | 2017-12-12 | 安徽农业大学 | A kind of method of Acer saccharum tissue culture quick breeding |
| CN114711143A (en) * | 2022-04-21 | 2022-07-08 | 东北林业大学 | Method for asexually and rapidly propagating acer truncatum seedlings through tissue culture |
| CN116267604A (en) * | 2023-02-18 | 2023-06-23 | 西北农林科技大学 | Method for obtaining acer truncatum aseptic seedlings and directly inducing aseptic buds |
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| CN104488703A (en) * | 2014-11-25 | 2015-04-08 | 巴中七彩林业科技有限公司 | Acer negundo aurea tissue culture rapid propagation method |
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| CN104488703A (en) * | 2014-11-25 | 2015-04-08 | 巴中七彩林业科技有限公司 | Acer negundo aurea tissue culture rapid propagation method |
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| 李艳敏等: "金叶复叶械组织培养技术研究", 《河南农业科学》 * |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106305427A (en) * | 2016-08-30 | 2017-01-11 | 长江三峡生态园林有限公司 | Tissue culture and rapid propagation method for Acer ginnala |
| CN106305427B (en) * | 2016-08-30 | 2018-04-10 | 长江三峡生态园林有限公司 | A kind of method of Amur maple tissue-culturing quick-propagation |
| CN107455261A (en) * | 2017-09-15 | 2017-12-12 | 安徽农业大学 | A kind of method of Acer saccharum tissue culture quick breeding |
| CN114711143A (en) * | 2022-04-21 | 2022-07-08 | 东北林业大学 | Method for asexually and rapidly propagating acer truncatum seedlings through tissue culture |
| CN114711143B (en) * | 2022-04-21 | 2023-08-08 | 东北林业大学 | Method for asexually and rapidly propagating Acer ginnala Maxim seedlings through tissue culture |
| CN116267604A (en) * | 2023-02-18 | 2023-06-23 | 西北农林科技大学 | Method for obtaining acer truncatum aseptic seedlings and directly inducing aseptic buds |
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