CN102577968A - Adventitious bud induction method for tree peony - Google Patents
Adventitious bud induction method for tree peony Download PDFInfo
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- CN102577968A CN102577968A CN2012100567643A CN201210056764A CN102577968A CN 102577968 A CN102577968 A CN 102577968A CN 2012100567643 A CN2012100567643 A CN 2012100567643A CN 201210056764 A CN201210056764 A CN 201210056764A CN 102577968 A CN102577968 A CN 102577968A
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- tree peony
- adventitious bud
- explant
- naa
- inducing method
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Abstract
The invention relates to an adventitious bud induction method for tree peony, which takes NAA and 6-BA as inducer and receptacle as explant to perform induced cultivation. The method takes the naphthylacetic acid (NAA) and 6-benzyl purine (6-BA) as the inducer, and takes the receptacle as the explant, the receptacle is abundant in nutrient content, and high in enzymes activity, the regeneration rate of the adventitious bud is higher, and can reach 100-490.91%, and the vitrifaction phenomenon is not serious, so that the scale production or the gene transformation can be carried out.
Description
Background technology
The present invention relates to the breeding method of a kind of tree peony, relate in particular to a kind of tree peony adventitious bud inducing method.
Technical field
Tree peony is China's national flower, and large flower and brilliant color is elegant and poised, also generally like for common people, but tree peony mainly leans on plant division or propagation by grafiting, and reproduction coefficient is low, and reproduction speed is slow, can not satisfy actual needs.At present, for the fast numerous research of tree peony, majority is that employing stem apex or sleeping bud are explant; This method through cell dedifferentiation and differentiation again, does not belong to micro cuttage, and reproduction coefficient is low; The seedling that obtains is mainly low because of take root difficulty and transplanting survival rate, is difficult to apply aborning.
Existing document adopts the approach of adventitious shoot regeneration, but explant is generally selected mature embryo, seed or seedling for use.Li Yuhua (2005) has set up tree peony ' Feng Dan is white ' regeneration of cotyledons system; Xiao Xiaopeng (2008) is that explant has successfully been set up fast traditional font system with the manyflower melastoma herb seed; Liu Lei (2009) has set up the complete regenerating system of tree peony ' Feng Dan is white ' mature embryo through callus approach differentiation indefinite bud, and aforesaid way all derives from the regenerating system of seed, has variation, can not keep the characteristic of the original kind of tree peony.
Beruto, it is explant that et al (2004) adopts peony petal and filigree, and only the callus of petal or petal generation is induced the regeneration that has obtained indefinite bud down at TDZ, and this kind mode vitrification phenomenon is more serious, is difficult to form the indefinite bud of completion.
Summary of the invention
The object of the present invention is to provide the high tree peony adventitious bud inducing method of a kind of regeneration rate.
To achieve these goals, technical scheme of the present invention has adopted a kind of tree peony adventitious bud inducing method, is derivant with NAA and 6-BA, is that explant carries out induction culturing with the holder.
Tree peony adventitious bud inducing method of the present invention specifically may further comprise the steps: with the tender bud of tree peony children, remove sepal after, with 70% alcohol-pickled 15-25sec, clean once, after soak soaks 8-20min, aseptic washing 3-5 time, 2-4min at every turn; Inoculate as explant with holder, be inoculated into the MS medium and add 1.5-2.5mgL
-16-benzyl purine 6-BA+0.1-0.3mgL
-1Methyl NAA, sucrose 25-35gL
-1, agar 7-8gL
-1, pH transfers to 5.8, illumination every day 14h, and intensity of illumination 3000lux, 25 ℃ of temperature, fresh culture is gone in switching in per 30 days.
Described fresh culture is to have added 1.5-2.5mgL in the MS medium
-16-benzyl purine 6-BA+0.1-0.3mgL
-1Methyl NAA, sucrose 25-35gL
-1, agar 7-8gL
-1
Described cleaning is cleaned for adopting sterile water.
Described soak is 0.1% HgCl
2Or 2% clorox.
The present invention adopts methyl (NAA) and 6-benzyl purine (6-BA) for inducing, and is explant with the holder, and the holder nutrient content is abundant; Enzyme is active high, and the adventitious shoot regeneration rate is higher, can reach 100-490.91%; And vitrification phenomenon is not serious, can modelling production or genetic transformation.
Embodiment
Embodiment 1
The method concrete steps of present embodiment are: select the tender bud of tree peony children, remove sepal after, 70% alcohol-pickled 20sec, aseptic then washing once, the HgCl through 0.1%
2After soaking 10min, aseptic washing 4 times, each 3min.Put on the blotting paper and be divided into 4 parts to the holder of pie, inoculate as explant; Be inoculated into the MS medium and add 2.0mgL-16-BA+0.2mgL-1NAA, sucrose 30gL-1, agar 7.5gL-1, pH transfers to 5.8; Illumination every day 14h, intensity of illumination 3000lux, 25 ℃ of temperature; Fresh culture is gone in switching in per 30 days, and through the inducing culture in 7 weeks, the adventitious bud inducing production rate of part tree peony kind reaches 100%.
Embodiment 2
The method concrete steps of present embodiment are: with the tender bud of tree peony children, remove sepal after,, clean once the HgCl of warp 0.1% with 70% alcohol-pickled 15sec
2After soaking 15min, aseptic washing 3 times, each 2min; Inoculate as explant with holder, be inoculated into the MS medium and add 1.5mgL
-16-benzyl purine 6-BA+0.1mgL
-1Methyl NAA, sucrose 25gL
-1, agar 7gL
-1, pH transfers to 5.8, illumination every day 14h, and intensity of illumination 3000lux, 25 ℃ of temperature, fresh culture is gone in switching in per 30 days.Through the inducing culture in 7 weeks, the adventitious bud inducing production rate of part tree peony kind reaches 330%.
Embodiment 3
The method concrete steps of present embodiment are: with the tender bud of tree peony children, remove sepal after, with 70% alcohol-pickled 25sec, clean once, after soak soaks 20min, aseptic washing 5 times, 4min at every turn; Inoculate as explant with holder, be inoculated into the MS medium and add 2.5mgL
-16-benzyl purine 6-BA+0.3mgL
-1Methyl NAA, sucrose 35gL
-1, agar 8gL
-1, pH transfers to 5.8, illumination every day 14h, and intensity of illumination 3000lux, 25 ℃ of temperature, fresh culture is gone in switching in per 30 days.Through the inducing culture in 7 weeks, the adventitious bud inducing production rate of part tree peony kind reaches 490.91%.
Claims (5)
1. tree peony adventitious bud inducing method, it is characterized in that: with NAA and 6-BA is derivant, is that explant carries out induction culturing with the holder.
2. tree peony adventitious bud inducing method according to claim 1 is characterized in that: may further comprise the steps: with the tender bud of tree peony children, remove sepal after; With 70% alcohol-pickled 15-25sec, clean once, after soak soaks 8-20min; Aseptic washing 3-5 time, each 2-4min; Inoculate as explant with holder, be inoculated into the MS medium and add 1.5-2.5mgL
-16-benzyl purine 6-BA+0.1-0.3mgL
-1Methyl NAA, sucrose 25-35gL
-1, agar 7-8gL
-1, pH transfers to 5.8, illumination every day 14h, and intensity of illumination 3000lux, 25 ℃ of temperature, fresh culture is gone in switching in per 30 days.
3. tree peony adventitious bud inducing method according to claim 2 is characterized in that: described fresh culture is to have added 1.5-2.5mgL in the MS medium
-16-benzyl purine 6-BA+0.1-0.3mgL
-1Methyl NAA, sucrose 25-35gL
-1, agar 7-8gL
-1
4. tree peony adventitious bud inducing method according to claim 2 is characterized in that: described cleaning is cleaned for adopting sterile water.
5. tree peony adventitious bud inducing method according to claim 2 is characterized in that: described soak is 0.1% HgCl
2Or 2% clorox.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN 201210056764 CN102577968B (en) | 2012-03-06 | 2012-03-06 | Adventitious bud induction method for tree peony |
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| Application Number | Priority Date | Filing Date | Title |
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| CN 201210056764 CN102577968B (en) | 2012-03-06 | 2012-03-06 | Adventitious bud induction method for tree peony |
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| CN102577968A true CN102577968A (en) | 2012-07-18 |
| CN102577968B CN102577968B (en) | 2013-10-16 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103120128A (en) * | 2013-02-28 | 2013-05-29 | 河南科技大学 | Method for avoiding paeonia suffruticosa tissue culture seedling browning and vitrifying |
| CN103270949A (en) * | 2013-05-22 | 2013-09-04 | 河南省农业科学院 | Novel peony tissue culture rooting method |
| CN104285787A (en) * | 2014-09-12 | 2015-01-21 | 河南科技学院 | Method for inducing peony receptacle callus tissues and differentiating adventitious buds |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102144563A (en) * | 2009-05-11 | 2011-08-10 | 中国林业科学研究院林业研究所 | Method for inducing calluses of peony anther |
| CN102172220A (en) * | 2009-05-11 | 2011-09-07 | 中国林业科学研究院林业研究所 | Method for inducing calluses of stems and leaves of tree peony |
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2012
- 2012-03-06 CN CN 201210056764 patent/CN102577968B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102144563A (en) * | 2009-05-11 | 2011-08-10 | 中国林业科学研究院林业研究所 | Method for inducing calluses of peony anther |
| CN102172220A (en) * | 2009-05-11 | 2011-09-07 | 中国林业科学研究院林业研究所 | Method for inducing calluses of stems and leaves of tree peony |
Non-Patent Citations (2)
| Title |
|---|
| 何宽远: "菊花花瓣与花托再生体系的建立", 《农技服务》, vol. 27, no. 5, 15 May 2010 (2010-05-15), pages 640 * |
| 高昌勇: "不同牡丹外植体诱导愈伤组织的研究", 《安徽农业科学》, vol. 35, no. 34, 10 December 2007 (2007-12-10), pages 11036 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103120128A (en) * | 2013-02-28 | 2013-05-29 | 河南科技大学 | Method for avoiding paeonia suffruticosa tissue culture seedling browning and vitrifying |
| CN103270949A (en) * | 2013-05-22 | 2013-09-04 | 河南省农业科学院 | Novel peony tissue culture rooting method |
| CN103270949B (en) * | 2013-05-22 | 2015-03-11 | 河南省农业科学院 | Novel peony tissue culture rooting method |
| CN104285787A (en) * | 2014-09-12 | 2015-01-21 | 河南科技学院 | Method for inducing peony receptacle callus tissues and differentiating adventitious buds |
| CN104285787B (en) * | 2014-09-12 | 2017-05-17 | 河南科技学院 | Method for inducing peony receptacle callus tissues and differentiating adventitious buds |
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| Publication number | Publication date |
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| CN102577968B (en) | 2013-10-16 |
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Inventor after: Wu Zhengjing Inventor after: Wu Zufeng Inventor after: Li Dongsheng Inventor after: Gao Wen Inventor after: Liu Yanan Inventor after: Du Xiaohong Inventor after: Hu Canli Inventor after: Jin Xiangli Inventor after: Ren Zhimin Inventor before: Wu Zhengjing Inventor before: Wu Zufeng Inventor before: Li Dongsheng Inventor before: Gao Wen Inventor before: Liu Yanan Inventor before: Du Xiaohong Inventor before: Hu Canli Inventor before: Jin Xiangli Inventor before: Ren Zhimin |
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