CN103636492B - Rhododendron hybrides cosmopolitan tissue culture rapid propagation method - Google Patents
Rhododendron hybrides cosmopolitan tissue culture rapid propagation method Download PDFInfo
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- CN103636492B CN103636492B CN201310563498.8A CN201310563498A CN103636492B CN 103636492 B CN103636492 B CN 103636492B CN 201310563498 A CN201310563498 A CN 201310563498A CN 103636492 B CN103636492 B CN 103636492B
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Abstract
The invention relates to a rhododendron hybrides cosmopolitan tissue culture rapid propagation method which comprises the following steps: A. explant treatment; B. culture initiation; C. multiple shoot culture; D. strong seedling culture, that is, transferring and inoculating aseptic seedlings obtained by the multiple shoot culture to a strong seedling culture medium, and culturing for 75 days at a day temperature of 25+/-1 DEG C, a night temperature of 22+/-1 DEG C, and an illumination intensity of 50 [mu]mol.m<-2>.s<-1> and with illumination for 12 h/day; E. rooting culture, that is, culturing for 90 days at a day temperature of 25+/-1 DEG C, a night temperature of 22+/-1 DEG C, and an illumination intensity of 50 [mu]mol.m<-2>.s<-1> and with illumination for 12 h/day. The method of the invention is not influenced by time and changes of season or environment, greatly increases the propagation rate, and improves the ornamental value and commercial value of rhododendron hybrides cosmopolitan.
Description
Technical field
The present invention relates to a kind of alpine rose flour gold butterfly tissue culture quick propagation culturing method, belong to field of plant growing technology, be mainly used in breeding in enormous quantities of alpine rose new varieties.
Background technology
Alpine rose flour gold butterfly (Rhododendron hybrides " Cosmopolitan ") is the cuculiform hybrid cultivar of Cuculidae, evergreen shrubs and dungarunga, pollen is red, and edge takes off for white powder, with the great red spot of a brown, plant is dense, and plant type is unfolded, leaf green light, cold-resistant and heat-resisting, do not fall leaves for-23 DEG C, premunition is strong, the ornamental value that tool is high.The propagation method such as alpine rose propagation method application seed propagation, cottage propagation, propagation by layering, propagation by grafiting, but these technology limit by several factors such as season, maternal plant materials, and survival rate is extremely low, is difficult to carry out large-scale production.Alpine rose tissue cultures only relates to several kinds such as delavay rhododendron (Rhododendron delavayi Franch), capitate rhododendron branchlet and leaf (RhododendronParvifolium Adoms), Rhododendron aureum georgi (Rhododendron Chrysantiuum pall), not yet reports the research of the whole breeding system of alpine rose flour gold butterfly tissue cultures both at home and abroad.
Summary of the invention
The object of the present invention is to provide a kind of not by the alpine rose flour gold butterfly method for tissue culture of extraneous natural environment restriction, to improve proliferative speed and commercial value type.
This alpine rose flour gold butterfly tissue culture quick propagation culturing method, comprises the steps:
A, explant process: choosing healthy and strong harmless alpine rose flour gold butterfly stem with bud is explant, first uses the alcohol-pickled 30s of 75%, after aseptic water washing 3 times, then carry out sterilizing 3min with 0.1% mercuric chloride, aseptic water washing 3 times;
B, Primary culture: the explant of sterilization treatment is inoculated into Primary culture base, day temperature is 25 ± 1 DEG C, night 22 ± 1 DEG C, and intensity of illumination is 50 μm of olm-2s-1, illumination every day 12h, incubation time 30 days;
Above-mentioned Primary culture base for mother liquor, sees attached list 1 with Read medium.Add IBA0.2mg/L, TDZ0.5mg/L, sucrose 25mg/L, agar 6.5mg/L, adjust pH=5.3, for subsequent use after 121 DEG C of autoclaving 20min.
C, adventitious shoots culture: the aseptic seedling that Primary culture grows is transferred to adventitious shoots culture base, and day temperature is 25 ± 1 DEG C, night 22 ± 1 DEG C, and intensity of illumination is 50 μm of olm-2s-1, illumination every day 12h, incubation time 75 days;
D, strong seedling culture: the aseptic seedling of adventitious shoots culture is transferred to strong seedling culture base, day temperature is 25 ± 1 DEG C, night 22 ± 1 DEG C, and intensity of illumination is 50 μm of olm-2s-1, illumination every day 12h, incubation time 75 days;
E, culture of rootage: day temperature is 25 ± 1 DEG C, night 22 ± 1 DEG C, intensity of illumination is 50 μm of olm-2s-1, illumination every day 12h, incubation time 90 days.
Described alpine rose flour gold butterfly tissue culture quick propagation culturing method, described in step D, strong seedling culture base is with WPM medium for mother liquor, sees attached list 2.Add sucrose 25mg/L, agar 6.5mg/L, adjust pH=5.3, for subsequent use after 121 DEG C of autoclaving 20min.
Described alpine rose flour gold butterfly tissue culture quick propagation culturing method, the root media described in step e for mother liquor, sees attached list 3 with Anderson medium.Add NAA0.2mg/L, IBA0.2mg/L, sucrose 25mg/L, agar 6.5mg/L, pH=5.3, for subsequent use after 121 DEG C of autoclaving 20min.
The present invention and utilize tissue culture propagation to study alpine rose flour gold butterfly, by the time and season environment change, and greatly improve proliferative speed, improve ornamental value and the commercial value of alpine rose flour gold butterfly.Protection quality germplasm, excavation are created to new germ plasm and had great significance, and application prospect is very wide.
Embodiment
Method step of the present invention is described in detail below in conjunction with instantiation,
Described alpine rose flour gold butterfly kind is introduced from Ghent, Belgium in November, 09, and afterwards nearly 4 years of Shijiazhuang City agricultural and forest science research institute domestication, this alpine rose flour gold butterfly tissue culture quick propagation culturing method, comprises the steps:
A, explant process: choosing healthy and strong harmless alpine rose flour gold butterfly stem with bud is explant, first uses the alcohol-pickled 30s of 75%, after aseptic water washing 3 times, then carry out sterilizing 3min with 0.1% mercuric chloride, aseptic water washing 3 times;
B, Primary culture: the explant of sterilization treatment is inoculated into Primary culture base, day temperature is 25 ± 1 DEG C, night 22 ± 1 DEG C, and intensity of illumination is 50 μm of olm-2s-1, illumination every day 12h, incubation time 30 days;
Above-mentioned Primary culture base for mother liquor, sees attached list 1 with Read medium.Add IBA0.2mg/L, TDZ0.5mg/L, sucrose 25mg/L, agar 6.5mg/L, adjust pH=5.3, for subsequent use after 121 DEG C of autoclaving 20min.
C, adventitious shoots culture: the aseptic seedling that Primary culture grows is transferred to adventitious shoots culture base, and day temperature is 25 ± 1 DEG C, night 22 ± 1 DEG C, and intensity of illumination is 50 μm of olm-2s-1, illumination every day 12h, incubation time 75 days;
Above-mentioned adventitious shoots culture base for mother liquor, sees attached list 2 with WPM medium.Add 2ip2.5mg/L, NAA0.1mg/L, sucrose 25mg/L, agar 6.5mg/L, adjust pH=5.3, for subsequent use after 121 DEG C of autoclaving 20min.
D, strong seedling culture: the aseptic seedling of adventitious shoots culture is transferred to strong seedling culture base, day temperature is 25 ± 1 DEG C, night 22 ± 1 DEG C, and intensity of illumination is 50 μm of olm-2s-1, illumination every day 12h, incubation time 75 days;
Above-mentioned strong seedling culture base for mother liquor, sees attached list 2 with WPM medium.Add sucrose 25mg/L, agar 6.5mg/L, adjust pH=5.3, for subsequent use after 121 DEG C of autoclaving 20min.
E, culture of rootage: day temperature is 25 ± 1 DEG C, night 22 ± 1 DEG C, intensity of illumination is 50 μm of olm-2s-1, illumination every day 12h, incubation time 90 days.
Above-mentioned root media for mother liquor, sees attached list 3 with Anderson medium.Add NAA0.2mg/L, IBA0.2mg/L, sucrose 25mg/L, agar 6.5mg/L, pH=5.3, for subsequent use after 121 DEG C of autoclaving 20min.
Subordinate list 1:Read medium mother liquor
Attention: the water content examining various chemical composition before preparation
Subordinate list 2:WPM medium mother liquor
Attention: the water content examining various chemical composition before preparation
Subordinate list 3:Anderson medium mother liquor
Attention: the water content examining various chemical composition before preparation
Claims (1)
1. an alpine rose flour gold butterfly tissue culture quick propagation culturing method, comprises the steps:
A, explant process: choosing healthy and strong harmless alpine rose flour gold butterfly stem with bud is explant, first uses the alcohol-pickled 30s of 75%, after aseptic water washing 3 times, then carry out sterilizing 3min with 0.1% mercuric chloride, aseptic water washing 3 times;
B, Primary culture: the explant of sterilization treatment is inoculated into Primary culture base, day temperature is 25 ± 1 DEG C, night 22 ± 1 DEG C, and intensity of illumination is 50 μm of olm
-2s
-1, illumination every day 12h, incubation time 30 days; Described Primary culture base for mother liquor, sees attached list 1:Read medium mother liquor with Read medium
Note adding IBA 0.2mg/L, TDZ0.5mg/L, sucrose 25mg/L, agar 6.5mg/L, adjust pH=5.3, for subsequent use after 121 DEG C of autoclaving 20min;
C, adventitious shoots culture: the aseptic seedling that Primary culture grows is transferred to adventitious shoots culture base, and day temperature is 25 ± 1 DEG C, night 22 ± 1 DEG C, and intensity of illumination is 50 μm of olm
-2s
-1, illumination every day 12h, incubation time 75 days; Described adventitious shoots culture base for mother liquor, sees attached list 2:WPM medium mother liquor with WPM medium
Add 2ip2.5mg/L, NAA0.1mg/L, sucrose 25mg/L, agar 6.5mg/L, adjust pH=5.3, for subsequent use after 121 DEG C of autoclaving 20min;
D, strong seedling culture: the aseptic seedling of adventitious shoots culture is transferred to strong seedling culture base, day temperature is 25 ± 1 DEG C, night 22 ± 1 DEG C, and intensity of illumination is 50 μm of olm
-2s
-1, illumination every day 12h, incubation time 75 days; Described strong seedling culture base for mother liquor, adds sucrose 25mg/L, agar 6.5mg/L with WPM medium, adjusts pH=5.3, for subsequent use after 121 DEG C of autoclaving 20min;
E, culture of rootage: day temperature is 25 ± 1 DEG C, night 22 ± 1 DEG C, intensity of illumination is 50 μm of olm
-2s
-1, illumination every day 12h, incubation time 90 days; Described root media for mother liquor, sees attached list 3:Anderson medium mother liquor with Anderson medium
Add NAA0.2mg/L, IBA 0.2mg/L, sucrose 25mg/L, agar 6.5mg/L, pH=5.3, for subsequent use after 121 DEG C of autoclaving 20min.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105850737B (en) * | 2016-04-08 | 2018-01-19 | 石家庄市农林科学研究院 | A kind of alpine rose " young beauty " chromosome doubling method |
| CN107242139B (en) * | 2017-08-15 | 2024-04-19 | 石家庄市农林科学研究院 | Induction method of rhododendron alpine 'Pink butterfly' somatic embryo |
| CN107494274B (en) * | 2017-10-12 | 2019-10-22 | 云南省农业科学院花卉研究所 | A kind of stripping bud sterilization method of alpine rose explant |
| CN109349109A (en) * | 2018-11-23 | 2019-02-19 | 云南省农业科学院花卉研究所 | Shaping methods in the bottle of alpine rose young beauty |
| CN109496854A (en) * | 2018-11-29 | 2019-03-22 | 四川农业大学 | A kind of pediment cuckoo seed tissue culture and rapid propagation method |
| CN109937885A (en) * | 2019-05-07 | 2019-06-28 | 贵州师范大学 | A kind of sterile seedling fast reproducing method of wrinkle leaf cuckoo |
| CN113100060B (en) * | 2021-04-20 | 2022-08-05 | 重庆市风景园林科学研究院 | Tissue culture propagation method for alpine rhododendron |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1568670A (en) * | 2004-04-26 | 2005-01-26 | 上海市闵行区农业科学研究所 | Tissue culture and tachytelic propagating method for alpine rhododendron |
| CN101984787A (en) * | 2010-11-10 | 2011-03-16 | 天津滨海国际花卉科技园区股份有限公司 | Method for strengthening tissue culture seedlings of rhododendron lapponicum |
| CN102090262A (en) * | 2010-12-13 | 2011-06-15 | 江苏省农业科学院 | Method for grafting and propagating rhododendron hybrids |
| CN102919133A (en) * | 2012-12-03 | 2013-02-13 | 江苏省农业科学院 | Technical method for promoting tissue culture rooting of alpine rose amethyst |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| JP2970277B2 (en) * | 1992-12-25 | 1999-11-02 | 王子製紙株式会社 | A method for promoting rooting of cultured shoots in Rhododendron plants |
| JPH06189643A (en) * | 1992-12-25 | 1994-07-12 | New Oji Paper Co Ltd | Large-scale multiplication of rhododendron |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1568670A (en) * | 2004-04-26 | 2005-01-26 | 上海市闵行区农业科学研究所 | Tissue culture and tachytelic propagating method for alpine rhododendron |
| CN101984787A (en) * | 2010-11-10 | 2011-03-16 | 天津滨海国际花卉科技园区股份有限公司 | Method for strengthening tissue culture seedlings of rhododendron lapponicum |
| CN102090262A (en) * | 2010-12-13 | 2011-06-15 | 江苏省农业科学院 | Method for grafting and propagating rhododendron hybrids |
| CN102919133A (en) * | 2012-12-03 | 2013-02-13 | 江苏省农业科学院 | Technical method for promoting tissue culture rooting of alpine rose amethyst |
Non-Patent Citations (1)
| Title |
|---|
| 不同基因型高山杜鹃茎段离体培养及工厂化繁育体系优化;姜泽盛等;《山东农业科学》;20130228;第45卷(第2期);第56页第3段 * |
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Effective date of registration: 20180116 Address after: 050041 Shijiazhuang Shengli Street, No. 479, Hebei Co-patentee after: SHIJIAZHUANG SHENZHOU FLOWER RESEARCH INSTITUTE CO., LTD. Patentee after: Shijiazhuang Academy of Agriculture and Forestry Sciences Address before: 050021 Cang Xing street, Shijiazhuang, Hebei Province, No. 16 Co-patentee before: SHIJIAZHUANG SHENZHOU FLOWER RESEARCH INSTITUTE Patentee before: Shijiazhuang Academy of Agriculture and Forestry Sciences |
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