CN102172220A - Method for inducing calluses of stems and leaves of tree peony - Google Patents
Method for inducing calluses of stems and leaves of tree peony Download PDFInfo
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- CN102172220A CN102172220A CN 201110046908 CN201110046908A CN102172220A CN 102172220 A CN102172220 A CN 102172220A CN 201110046908 CN201110046908 CN 201110046908 CN 201110046908 A CN201110046908 A CN 201110046908A CN 102172220 A CN102172220 A CN 102172220A
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Abstract
The invention discloses a method for producing calluses of stems and leaves of a tree peony. In the method, the calluses are induced by using an MS (Murashige and Skoog) culture medium; and a 6-BA (Benzyl Aminopurine) cytokinin, an NAA (Naphthyl Acetic Acid) plant growth regulator, a 2,4-D herbicide and other materials are mixed in the MS culture medium. Specific culture medium schemes and hormone constitutions are provided for explants, such as stem sections and leaves of the tree peony; and the optimal material obtaining period of the calluses is elaborated. Because the inducing rates of the calluses of the tree peony are both more than 80%, a better inducing effect is realized.
Description
The application divides an application for following application:
Application number: 200910083744.3; Denomination of invention: tree peony callus induction method; The applying date: 2009.5.11.
Technical field
The present invention relates to a kind of callus culture method, relate in particular to a kind of tree peony cauline leaf callus induction method.
Background technology
Tree peony is one of China's ten big famous flowers, also is the national flower of China, and the good reputation of " king in spending " is arranged.The tree peony tissue culture mainly is to utilize bud to induce plant and utilize explant to induce by the callus approach, and the callus induction of tree peony is an important stage for regeneration plant, higher callus induction rate lays a good foundation for the later stage differentiation of calli, and how obtaining the kind of higher callus induction rate and type of culture medium, hormone and concentration and condition of culture has confidential relation.
Callus culture is a kind of modal cultivation form, and except that shoot apical meristem cultivation and a part of organ culture, other several plant tissue culture forms finally all will experience callus could produce plant.Callus also usually is the source of suspension cultured cells and protoplast in addition.Callus is divided into embryo callus and non-embryonic callus tissue, and embryo callus is meant the callus of somatic embryo, through being divided into the tender plant of children.
In the prior art, the tree peony Study on tissue culture has had long history, research contents also mainly concentrates on fields such as explant selection, growth regulatory substance proportioning and culture medium prescription, mainly be to utilize stem apex direct development to become plant, and utilize the explant induction callus, utilize the research of callus differentiation generation plant less relatively then, and inducing of callus is the precondition that the callus differentiation produces plant.
There is following shortcoming at least in above-mentioned prior art:
There are callus induction difficulty, rooting rate and problems such as difficulty, brownization, vitrifying are transplanted in of low quality, the domestication of taking root.
Summary of the invention
The purpose of this invention is to provide the high tree peony cauline leaf callus induction method of a kind of callus induction rate.
The objective of the invention is to be achieved through the following technical solutions:
Tree peony cauline leaf callus induction method of the present invention, comprise and choose the tree peony explant that adopt the MS medium to carry out inducing of callus, described tree peony explant comprises stem, be the period of drawing materials: at the beginning of 4 months by the end of March spring, get tender stem section of children or the young tender stem section that is formed by seed sprouting;
The MS medium of stem comprises: MS+NAA0.05mg/L+6-BA2.0mg/L+2,4-D1.0mg/L.
Described tree peony explant can also comprise blade, and be the period of drawing materials: at the beginning of 4 months by the end of March spring, get tender blade of children or the young tender blade that is formed by seed sprouting;
The MS medium of blade comprises: MS+6-BA0.1mg/L+2,4-D0.5mg/L.
As seen from the above technical solution provided by the invention, tree peony cauline leaf callus induction method of the present invention, owing to adopt the MS medium to carry out inducing of tree peony cauline leaf callus, be furnished with the 6-BA basic element of cell division, NAA plant growth regulator, 2 in the MS medium, materials such as 4-D weed killer herbicide.Choose the tree peony explant in suitable period and suitable medium, the callus induction rate height.
Embodiment
Tree peony cauline leaf callus induction method of the present invention, its preferable embodiment are to comprise and choose the tree peony explant that employing MS medium carries out inducing of callus, is furnished with following one or more materials in the described MS medium:
The 6-BA basic element of cell division, NAA plant growth regulator, 2, the 4-D weed killer herbicide.
Before carrying out the inducing of callus, can cultivate pre-treatment earlier:
At first, selected tree peony explant is carried out 4 ℃ of low temperature treatment;
Then, utilize 2%NaClO sterilization 30min, 70% alcohol disinfecting 30s, aseptic water washing 3-5 time.
Specific embodiment:
With different explants is example, adopts different medium and different condition of culture to reach the best effect of inducing.Be that object is set forth respectively below with the different explants:
Inducing of embryo callus:
Cultivation period: annual September, after seed forms;
Cultivate pre-treatment: utilize the method that low temperature (4 ℃) is handled and gibberellin treatment combines, inoculate after utilizing conventional sterilization method (2%NaClO sterilize 30min, 70% alcohol disinfecting 30s, aseptic water washing 3-5 time) to sterilize;
Best medium: MS+NAA0.1mg/L+6-BA1.0mg/L;
Callus induction rate reaches 100%.
Stem section callus induction:
Draw materials period: get the tender stem section of children, at the beginning of 4 months by the end of March spring, the perhaps young tender stem section that forms by seed sprouting;
Cultivate pre-treatment: utilize conventional sterilizing methods (2%NaClO sterilize 30min, 70% alcohol disinfecting 30s, aseptic water washing 3-5 time) to carry out disinfection;
Best medium: MS+NAA0.05mg/L+6-BA2.0mg/L+2,4-D1.0mg/L;
Callus induction rate is more than 80%.
The blade callus induction:
Draw materials period: get the tender blade of children, at the beginning of 4 months by the end of March spring, the perhaps young tender blade that forms by seed sprouting;
Cultivate pre-treatment: utilize conventional sterilizing methods (2%NaClO sterilize 30min, 70% alcohol disinfecting 30s, aseptic water washing 3-5 time) to carry out disinfection;
Optimal medium: MS+6-BA0.1mg/L+2,4-D0.5mg/L;
About callus induction rate 80%.
The petal callus induction:
Draw materials period: in the monokaryon mid-term of flower pesticide;
Cultivate pre-treatment: after handling 8d under 4 ℃ of conditions, carrying out disinfection is seeded to medium to utilize conventional sterilizing methods (2%NaClO sterilize 30min, 70% alcohol disinfecting 30s, aseptic water washing 3-5 time);
Optimal medium: MS+2,4-D1.0mg/L+6-BA2mg/L+NAA0.1mg/L;
About callus induction rate 60%.
Anther callus is induced:
Draw materials period: the monokaryon at flower pesticide is drawn materials mid-term;
Cultivate pre-treatment: after handling 8d under 4 ℃ of conditions, carrying out disinfection is seeded to medium to utilize conventional sterilizing methods (2%NaClO sterilize 30min, 70% alcohol disinfecting 30s, aseptic water washing 3-5 time);
Optimal medium: MS+2,4-D2.0mg/L+6-BA1.5mg/L+NAA1.0mg/L, sucrose concentration are 6%;
The inductivity about 50% of callus.
Among the present invention,, clear and definite medium scheme and hormone combinations are arranged all at different explants.And to having carried out comprehensive elaboration the period of drawing materials of the best of callus.The inductivity of tree peony callus has reached and has induced effect preferably all more than 80%; Petal and anther callus are induced also and have been reached more than 50%.
The above; only for the preferable embodiment of the present invention, but protection scope of the present invention is not limited thereto, and anyly is familiar with those skilled in the art in the technical scope that the present invention discloses; the variation that can expect easily or replacement all should be encompassed within protection scope of the present invention.
Claims (3)
1. tree peony cauline leaf callus induction method, it is characterized in that, comprise and choose the tree peony explant, adopt the MS medium to carry out inducing of callus, described tree peony explant comprises stem, be the period of drawing materials: at the beginning of 4 months by the end of March spring, get tender stem section of children or the young tender stem section that is formed by seed sprouting;
The MS medium of stem comprises: MS+NAA0.05mg/L+6-BA2.0mg/L+2,4-D1.0mg/L.
2. tree peony cauline leaf callus induction method according to claim 1 is characterized in that described tree peony explant also comprises blade, and be the period of drawing materials: at the beginning of 4 months by the end of March spring, get tender blade of children or the young tender blade that is formed by seed sprouting;
The MS medium of blade comprises: MS+6-BA0.1mg/L+2,4-D0.5mg/L.
3. tree peony cauline leaf callus induction method according to claim 1 and 2 is characterized in that, comprises the cultivation pre-treatment:
At first, selected tree peony explant is carried out 4 ℃ of low temperature treatment;
Then, utilize 2%NaClO sterilization 30min, 70% alcohol disinfecting 30s, aseptic water washing 3-5 time.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110046908 CN102172220B (en) | 2009-05-11 | 2009-05-11 | Method for inducing calluses of stems and leaves of tree peony |
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| Application Number | Priority Date | Filing Date | Title |
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| CN 201110046908 CN102172220B (en) | 2009-05-11 | 2009-05-11 | Method for inducing calluses of stems and leaves of tree peony |
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| Application Number | Title | Priority Date | Filing Date |
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| CN 200910083744 Division CN101548647B (en) | 2009-05-11 | 2009-05-11 | Tree peony callus induction method |
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| CN102172220A true CN102172220A (en) | 2011-09-07 |
| CN102172220B CN102172220B (en) | 2012-09-26 |
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| CN 201110046908 Expired - Fee Related CN102172220B (en) | 2009-05-11 | 2009-05-11 | Method for inducing calluses of stems and leaves of tree peony |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102577968A (en) * | 2012-03-06 | 2012-07-18 | 河南科技大学 | Adventitious bud induction method for tree peony |
| 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 |
| CN103355165A (en) * | 2012-04-09 | 2013-10-23 | 上海植物园 | Culture method of peony embryonic callus as well as culture medium |
| CN105917979A (en) * | 2016-06-16 | 2016-09-07 | 成都森洁商贸有限公司 | Operation method for grafting peonies |
| CN107593446A (en) * | 2017-10-19 | 2018-01-19 | 寿县正阳油用牡丹种植专业合作社 | A kind of fast breeding method of the induction and regeneration of tree peony |
| CN109602006A (en) * | 2019-01-09 | 2019-04-12 | 山东贝世康生物科技有限公司 | A kind of culture method of purification and application containing tree peony Stem Cell Activity object |
| CN116602212A (en) * | 2023-06-06 | 2023-08-18 | 北京市农林科学院 | Paeonia plant in-vitro culture and proliferation method and special culture medium |
-
2009
- 2009-05-11 CN CN 201110046908 patent/CN102172220B/en not_active Expired - Fee Related
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102577968A (en) * | 2012-03-06 | 2012-07-18 | 河南科技大学 | Adventitious bud induction method for tree peony |
| CN102577968B (en) * | 2012-03-06 | 2013-10-16 | 河南科技大学 | Adventitious bud induction method for tree peony |
| CN103355165A (en) * | 2012-04-09 | 2013-10-23 | 上海植物园 | Culture method of peony embryonic callus as well as culture medium |
| CN103355165B (en) * | 2012-04-09 | 2015-04-15 | 上海植物园 | Culture method of peony embryonic callus as well as culture medium |
| 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 |
| CN105917979A (en) * | 2016-06-16 | 2016-09-07 | 成都森洁商贸有限公司 | Operation method for grafting peonies |
| CN107593446A (en) * | 2017-10-19 | 2018-01-19 | 寿县正阳油用牡丹种植专业合作社 | A kind of fast breeding method of the induction and regeneration of tree peony |
| CN109602006A (en) * | 2019-01-09 | 2019-04-12 | 山东贝世康生物科技有限公司 | A kind of culture method of purification and application containing tree peony Stem Cell Activity object |
| CN116602212A (en) * | 2023-06-06 | 2023-08-18 | 北京市农林科学院 | Paeonia plant in-vitro culture and proliferation method and special culture medium |
| CN116602212B (en) * | 2023-06-06 | 2024-05-03 | 北京市农林科学院 | Paeonia plant in-vitro culture and proliferation method and special culture medium |
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| Publication number | Publication date |
|---|---|
| CN102172220B (en) | 2012-09-26 |
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