CN104663435A - Method for tissue culture and rapid propagation of acer palmatum hogyoku - Google Patents
Method for tissue culture and rapid propagation of acer palmatum hogyoku Download PDFInfo
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- CN104663435A CN104663435A CN201510064320.8A CN201510064320A CN104663435A CN 104663435 A CN104663435 A CN 104663435A CN 201510064320 A CN201510064320 A CN 201510064320A CN 104663435 A CN104663435 A CN 104663435A
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Abstract
The invention discloses a method for tissue culture and rapid propagation of acer palmatum hogyoku, belonging to the technical field of plant tissue culture. The method for the tissue culture and rapid propagation of the acer palmatum hogyoku comprises the following steps: sterilizing an explant, and carrying out culture, proliferation culture and rooting culture. The method for the tissue culture and rapid propagation of the acer palmatum hogyoku has the advantages that a tissue culture and rapid propagation system is established, a used culture medium adopts a simple formula, the culture medium is low in cost, a culture process is simple and convenient, the acer palmatum hogyoku propagation coefficient is high, and technical support is provided for industrial seedling culture and deep processing of the acer palmatum hogyoku.
Description
Technical field
The invention belongs to field of plant tissue culture technique, relate to a kind of tissue culture method, be specifically related to a kind of tissue culture and rapid propagation method of precious jade.
Background technology
Precious jade (
acer palmatum ' Hogyoku ') be a mutation of Aceraceae Acer Acer palmatum, its tree-like grace, autumn, leaf was bright-coloured darkorange, spring and summer leaf be darker emerald green.Precious jade plant is strong, resistance, and branch is dense, and growth is very fast, and one-tenth plant height in age reaches 6m, is easy to pruning shaping.Precious jade is as the good Acer palmatum new varieties of visual effect, and its bottle green and autumn leaves uniqueness orange very attractive, has very high ornamental value, can be used for garden, Chi Pan, road center isolation strip make scape, park irrigation, urban look etc.But precious jade quantity is few, generally adopts propagation by grafiting, its growth cycle is long, and cuttage is not easy to take root, and reproduction speed is comparatively slow, and reproduction rate is low, is difficult to the demand meeting market, not easily obtains economic benefit.
Therefore, the reproductive efficiency how improving precious jade becomes a key technology of precious jade resource provisioning needs solution.Plant Tissue Breeding is the one rapidly and efficiently propagation method of plant large-scale production.At present about the tissue culture technology of Acer palmatum, as number of patent application 201310338739.9 relates to a kind of Novel method for tissue culture rapid propagation of acer paimatum, it comprises the following steps: S1. cuts explant: gather Acer palmatum tender stem segments, cuts into the stem section that 2 ~ 3cm is long; S2. sterilizing: explant is put into superclean bench, with alcohol and mercuric chloride sterilizing; S3. Primary culture: the explant after sterilizing is inoculated in Primary culture base and cultivates, Primary culture base is WPM+GA30.1mg/L+TDZ0.05mg/L+ sucrose 30g/L+ agar 5g/L; S4. culture of rootage: choosing stem length after Primary culture is the Acer palmatum stem with bud of 1 ~ 2cm, is transferred in root media and cultivates after being cut by axillalry bud, root media is 1/2WPM+NAA0.05mg/L+ sucrose 25g/L+ agar 5g/L.But the training of the group of precious jade does not have relevant report, therefore, be badly in need of, to the research of precious jade group culturation rapid propagating technology, setting up precious jade tissue-culturing rapid propagation system and obtaining precious jade aseptic seedling.
Summary of the invention
The present invention utilizes tissue culture technique to carry out precious jade explant and cultivates acquisition precious jade aseptic seedling, provide a kind of tissue culture and rapid propagation method of precious jade, group training used medium formula is simple, group training simple flow, reproduction coefficient is higher, for precious jade factorial seedling growth and deep processing thereof provide technical support.
The present invention adopts the tissue culture technique of plant, utilizes precious jade stem explants to induce axillary buds formation, then induces axillary buds formation adventitious buds proliferation, and cutting Multiple Buds is taken root.
Object of the present invention is achieved through the following technical solutions: a kind of tissue culture and rapid propagation method of precious jade, and it comprises the following steps:
(1) explant sterilization: get the then raw edible tender branch of precious jade without damage by disease and insect, remove blade, stem section is cut into 2 ~ 3cm stem with bud, be placed on superclean bench, with 75% alcohol sterilizing 6 ~ 10s, 0.1% mercuric chloride sterilizing 6 ~ 10min, aseptic water washing 4 ~ 6 times;
(2) Primary culture: be inoculated into by the stem with bud after sterilizing in Primary culture base, cultivates 2 ~ 3 weeks in culturing room, and axillalry bud extends 2 ~ 4cm;
(3) Multiplying culture: cut by axillalry bud and be inoculated in proliferated culture medium, cultivates 4 ~ 6 weeks in culturing room, and Multiple Buds generates, plant height 1.5 ~ 3cm, and growth coefficient is 2 ~ 3;
(4) culture of rootage: the Multiple Buds choosing robust growth, is cut into stem-segment with single bud and is transferred in root media, cultivates 3 ~ 4 weeks, obtain the precious jade aseptic seedling of taking root in culturing room.
Described Primary culture base is B6+0.1mg/LIAA+0.05mg/LTDZ+ sucrose 30g/L+ agar 5g/L, pH 6.3 ~ 6.7.
Described proliferated culture medium is B6+0.1mg/LNAA+0.1mg/LTDZ+0.8mg/L6-BA+50mg/LLH+50mg/L Coconut Juice.
Described root media is 1/4B6+0.02mg/LIAA+0.05mg/LNAA+0.01mg/LIBA.
Cultivating indoor conditions described in step (2), step (3) and step (4) is: intensity of illumination 2000 ~ 3000LX, light application time 14 ~ 18h/d, temperature 24 ~ 28 DEG C.
The present invention has following beneficial effect: the present invention establishes tissue-culturing rapid propagation system, the culture medium prescription used is simple, culture medium cost is low, cultivates simple flow, improves the efficiency of precious jade breeding, reproduction coefficient is higher is 2 ~ 3, can obtain the precious jade aseptic seedling that genetic character is consistent fast; The present invention utilizes tissue culture technique, carries out explant cultivation and obtains precious jade aseptic seedling, by the impact of seasonal climate change, natural calamity, for precious jade industrialization nursery and deep processing thereof provide technical support.
Accompanying drawing explanation
Fig. 1 is axillary bud sprouting in Primary culture of the present invention;
Fig. 2 is the Multiple Buds generated in Multiplying culture of the present invention;
Fig. 3 is the precious jade aseptic seedling that culture of rootage of the present invention obtains;
Fig. 4 is the root system of the precious jade aseptic seedling that culture of rootage of the present invention obtains.
Embodiment
Below in conjunction with embodiment, the present invention will be further described, and protection scope of the present invention is not limited to the following stated.
embodiment 1:a tissue culture and rapid propagation method for precious jade, it comprises the following steps:
(1) explant sterilization: get the then raw edible tender branch of precious jade without damage by disease and insect, remove blade, stem section is cut into 2cm stem with bud, be placed on superclean bench, with 75% alcohol sterilizing 6s, 0.1% mercuric chloride sterilizing 10min, aseptic water washing 6 times;
(2) Primary culture: the stem with bud after sterilizing is inoculated in Primary culture base, be 2000LX in intensity of illumination, light application time is 18h/d, temperature is cultivate 2 weeks in the culturing room of 24 DEG C, axillalry bud average elongation 2cm, wherein, Primary culture base is: B6+0.1mg/LIAA+0.05mg/LTDZ+ sucrose 30g/L+ agar 5g/L, pH 6.3;
(3) Multiplying culture: axillalry bud is cut and is inoculated in proliferated culture medium, cultivation indoor conditions remains unchanged, cultivate 4 weeks, Multiple Buds generates, strain mean height 1.5cm, growth coefficient is 2, and wherein, proliferated culture medium is B6+0.1mg/LNAA+0.1mg/LTDZ+0.8mg/L6-BA+50mg/LLH+ 50mg/L Coconut Juice;
(4) culture of rootage: the Multiple Buds choosing robust growth, being cut into stem-segment with single bud is transferred in root media, cultivation indoor conditions remains unchanged, cultivate 3 weeks, obtain the precious jade aseptic seedling of taking root, wherein, root media is: 1/4B6+ 0.02mg/LIAA+0.05mg/LNAA+0.01mg/LIBA's.
embodiment 2:a tissue culture and rapid propagation method for precious jade, it comprises the following steps:
(1) explant sterilization: get the then raw edible tender branch of precious jade without damage by disease and insect, remove blade, stem section is cut into 3cm stem with bud, be placed on superclean bench, with 75% alcohol sterilizing 10s, 0.1% mercuric chloride sterilizing 6min, aseptic water washing 4 times;
(2) Primary culture: the stem with bud after sterilizing is inoculated in Primary culture base, be 3000LX in intensity of illumination, light application time is 14h/d, temperature is cultivate 3 weeks in the culturing room of 28 DEG C, axillalry bud average elongation 4cm, wherein Primary culture base is: B6+0.1mg/LIAA+0.05mg/LTDZ+ sucrose 30g/L+ agar 5g/L, pH 6.7;
(3) Multiplying culture: axillalry bud is cut and is inoculated in proliferated culture medium, cultivation indoor conditions remains unchanged, cultivate 6 weeks, Multiple Buds generates, strain mean height 3.0cm, growth coefficient is 3, and wherein, proliferated culture medium is B6+0.1mg/LNAA+0.1mg/LTDZ+0.8mg/L6-BA+50mg/LLH+ 50mg/L Coconut Juice;
(4) culture of rootage: the Multiple Buds choosing robust growth, being cut into stem-segment with single bud is transferred in root media, cultivation indoor conditions remains unchanged, cultivate 4 weeks, obtain the precious jade aseptic seedling of taking root, wherein, root media is: 1/4B6+ 0.02mg/LIAA+0.05mg/LNAA+0.01mg/LIBA's.
embodiment 3:a tissue culture and rapid propagation method for precious jade, it comprises the following steps:
(1) explant sterilization: get the then raw edible tender branch of precious jade without damage by disease and insect, remove blade, stem section is cut into 3cm stem with bud, be placed on superclean bench, with 75% alcohol sterilizing 7s, 0.1% mercuric chloride sterilizing 8min, aseptic water washing 5 times;
(2) Primary culture: the stem with bud after sterilizing is inoculated in Primary culture base, be 2300LX in intensity of illumination, light application time is 16h/d, temperature is cultivate 16 days in the culturing room of 25 DEG C, axillalry bud average elongation 2.8cm, wherein, Primary culture base is: B6+0.1mg/LIAA+0.05mg/LTDZ+ sucrose 30g/L+ agar 5g/L, pH 6.5;
(3) Multiplying culture: axillalry bud is cut and is inoculated in proliferated culture medium, cultivation indoor conditions remains unchanged, cultivate 5 weeks, Multiple Buds generates, strain mean height 2.3cm, growth coefficient is 2.3, and wherein, proliferated culture medium is B6+0.1mg/LNAA+ 0.1mg/LTDZ+0.8mg/L6-BA+50mg/LLH+ 50mg/L Coconut Juice;
(4) culture of rootage: the Multiple Buds choosing robust growth, being cut into stem-segment with single bud is transferred in root media, cultivation indoor conditions remains unchanged, cultivate 24 days, obtain the precious jade aseptic seedling of taking root, wherein, root media is: 1/4B6+0.02mg/LIAA+0.05mg/LNAA+0.01mg/LIBA.
embodiment 4:a tissue culture and rapid propagation method for precious jade, it comprises the following steps:
(1) explant sterilization: get the then raw edible tender branch of precious jade without damage by disease and insect, remove blade, stem section is cut into 2.5cm stem with bud, be placed on superclean bench, with 75% alcohol sterilizing 8s, 0.1% mercuric chloride sterilizing 7min, aseptic water washing 6 times;
(2) Primary culture: the stem with bud after sterilizing is inoculated in Primary culture base, be 2600LX in intensity of illumination, light application time is 15h/d, temperature is cultivate 20 days in the culturing room of 27 DEG C, axillalry bud average elongation 3.6cm, wherein, Primary culture base is B6+0.1mg/LIAA+0.05mg/LTDZ+ sucrose 30g/L+ agar 5g/L, pH 6.6;
(3) Multiplying culture: axillalry bud is cut and is inoculated in proliferated culture medium, cultivation indoor conditions remains unchanged, cultivate 5 weeks, Multiple Buds generates, strain mean height 2.5cm, growth coefficient is 2.6, and wherein, proliferated culture medium is B6+0.1mg/LNAA+ 0.1mg/LTDZ+0.8mg/L6-BA+50mg/LLH+ 50mg/L Coconut Juice;
(4) culture of rootage: the Multiple Buds choosing robust growth, being cut into stem-segment with single bud is transferred in root media, cultivation indoor conditions remains unchanged, cultivate 25 days, obtain the precious jade aseptic seedling of taking root, wherein, root media is: 1/4B6+0.02mg/LIAA+0.05mg/LNAA+0.01mg/LIBA's.
Draw from embodiment 1 ~ 4: the present invention utilizes plant tissue culture technique, set up tissue-culturing rapid propagation system, culture medium prescription used is simple, and growth coefficient is 2 ~ 3, can obtain a large amount of precious jade aseptic seedling in the short time.
Claims (5)
1. a tissue culture and rapid propagation method for precious jade, is characterized in that, it comprises the following steps:
(1) explant sterilization: get the then raw edible tender branch of precious jade without damage by disease and insect, remove blade, stem section is cut into 2 ~ 3cm stem with bud, be placed on superclean bench, with 75% alcohol sterilizing 6 ~ 10s, 0.1% mercuric chloride sterilizing 6 ~ 10min, aseptic water washing 4 ~ 6 times;
(2) Primary culture: be inoculated into by the stem with bud after sterilizing in Primary culture base, cultivates 2 ~ 3 weeks in culturing room, and axillalry bud extends 2 ~ 4cm;
(3) Multiplying culture: cut by axillalry bud and be inoculated in proliferated culture medium, cultivates 4 ~ 6 weeks in culturing room, and Multiple Buds generates, plant height 1.5 ~ 3cm;
(4) culture of rootage: the Multiple Buds choosing robust growth, is cut into stem-segment with single bud and is transferred in root media, cultivates 3 ~ 4 weeks, obtain the precious jade aseptic seedling of taking root in culturing room.
2. the tissue culture and rapid propagation method of a kind of precious jade according to claim 1, is characterized in that, described Primary culture base is B6+0.1mg/LIAA+0.05mg/LTDZ+ sucrose 30g/L+ agar 5g/L, pH 6.3 ~ 6.7.
3. the tissue culture and rapid propagation method of a kind of precious jade according to claim 1, is characterized in that, described proliferated culture medium is B6+0.1mg/LNAA+0.1mg/LTDZ+0.8mg/L6-BA+50mg/LLH+50mg/L Coconut Juice.
4. the tissue culture and rapid propagation method of a kind of precious jade according to claim 1, is characterized in that, described root media is 1/4B6+0.02mg/LIAA+0.05mg/LNAA+0.01mg/LIBA.
5. the tissue culture and rapid propagation method of a kind of precious jade according to claim 1, it is characterized in that, cultivating indoor conditions described in step (2), step (3) and step (4) is: intensity of illumination 2000 ~ 3000LX, light application time 14 ~ 18h/d, temperature 24 ~ 28 DEG C.
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Cited By (3)
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| CN105409782A (en) * | 2015-12-30 | 2016-03-23 | 四川禾木本业农林科技有限公司 | Tissue culture fast propagation method of acer palmatum 'Ki-hachijo' |
| CN105638460A (en) * | 2015-12-30 | 2016-06-08 | 四川禾木本业农林科技有限公司 | Tissue-culture rapid propagation method for Acer palmatum Butterfly |
| CN108935105A (en) * | 2018-08-24 | 2018-12-07 | 云南省农业科学院药用植物研究所 | A kind of limacia sagittata tissue culture and rapid propagation method |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105409782A (en) * | 2015-12-30 | 2016-03-23 | 四川禾木本业农林科技有限公司 | Tissue culture fast propagation method of acer palmatum 'Ki-hachijo' |
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| CN105409782B (en) * | 2015-12-30 | 2017-10-20 | 四川七彩林业开发有限公司 | A kind of yellow eight zhang tissue culture and rapid propagation method of mountain red autumnal leaves |
| CN108935105A (en) * | 2018-08-24 | 2018-12-07 | 云南省农业科学院药用植物研究所 | A kind of limacia sagittata tissue culture and rapid propagation method |
| CN108935105B (en) * | 2018-08-24 | 2021-08-27 | 云南省农业科学院药用植物研究所 | Tissue culture and rapid propagation method for Tinospora sinensis |
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Address after: 636611 six Huai Shu Village, Dongyu Town, Nanjiang County, Bazhong, Sichuan Patentee after: Sichuan Qicai Forestry Co., Ltd. Address before: 636611 six Huai Shu Village, Dongyu Town, Nanjiang County, Bazhong, Sichuan Patentee before: SICHUAN QICAI FORESTRY INDUSTRY DEVELOPMENT CO., LTD. |
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Address after: 635600 Tissue Culture Workshop Building of Rare and Colorful Leaf Plant Varieties in Guangwu Mountain, Sanshe, Changtan Village, Zhengzheng Town, Nanjiang County, Bazhong City, Sichuan Province Patentee after: Sichuan Qicai Forestry Co., Ltd. Address before: 636611 six Huai Shu Village, Dongyu Town, Nanjiang County, Bazhong, Sichuan Patentee before: Sichuan Qicai Forestry Co., Ltd. |