CN102144563B - Method for inducing calluses of peony anther - Google Patents
Method for inducing calluses of peony anther Download PDFInfo
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- CN102144563B CN102144563B CN 201110046751 CN201110046751A CN102144563B CN 102144563 B CN102144563 B CN 102144563B CN 201110046751 CN201110046751 CN 201110046751 CN 201110046751 A CN201110046751 A CN 201110046751A CN 102144563 B CN102144563 B CN 102144563B
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- callus
- tree peony
- calluses
- anther
- peony
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Abstract
The invention discloses a method for inducing calluses of peony anther. A murashige and skoog (MS) culture medium is adopted to induce the calluses; and substances such as 6-benzylamino adenine (BA) cytokinin, a naphthylacetic acid (NAA) plant growth regulator, a 2,4-D herbicide and the like are prepared in the MS culture medium. The definite culture medium scheme and hormone combination are provided for a peony anther explant. The optimal material taking period of the calluses is presented. The inductivity of peony calluses is over 80 percent, and the better induction effect is achieved; and the inductivity of the calluses of the anther is over 50 percent.
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 flower pesticide 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 be utilize bud induce plant with utilize explant to induce through 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 kind and the concentration and the condition of culture that how to obtain higher callus induction rate and type of culture medium, hormone have confidential relation.
Callus culture is a kind of modal cultivation form, and except that the shoot apical meristem cultivation with 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 is to utilize stem apex direct development to become plant, and utilizes 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 flower pesticide callus induction method of a kind of callus induction rate.
The objective of the invention is to realize through following technical scheme:
Tree peony flower pesticide callus induction method of the present invention comprises and chooses the tree peony explant, adopts the MS medium to carry out inducing of callus, and said tree peony explant is a flower pesticide, draws materials period: the monokaryon at flower pesticide is drawn materials mid-term;
Said MS medium comprises: MS+2,4-D2.0mg/L+6-BA1.5mg/L+NAA1.0mg/L comprises that also concentration is 6% sucrose.
Technical scheme by the invention described above provides can be found out; Tree peony flower pesticide callus induction method of the present invention; Owing to adopt the MS medium to carry out inducing of tree peony anther 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.And the phase is drawn materials the callus induction rate height in due course.
Embodiment
Tree peony flower pesticide 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 said 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%NaCl0 sterilization 30min, 70% alcohol disinfecting 30s, aseptic water washing 3-5 time.
Specific embodiment:
With different explants is example, adopts different culture medium and different culture condition 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 low temperature (4 ℃) to handle the method that combines with gibberellin treatment, inoculate after utilizing conventional sterilization method (2%NaCl0 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 young tender stem section that perhaps forms by seed sprouting;
Cultivate pre-treatment: utilize conventional sterilizing methods (2%NaCl0 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 young tender blade that perhaps forms by seed sprouting;
Cultivate pre-treatment: utilize conventional sterilizing methods (2%NaCl0 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%NaCl0 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%NaCl0 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 to 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; Be merely the preferable embodiment of the present invention, but protection scope of the present invention is not limited thereto, any technical staff who is familiar with the present technique field is 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 (1)
1. a tree peony flower pesticide callus induction method is characterized in that, comprises and chooses the tree peony explant, adopts the MS medium to carry out inducing of callus, and said tree peony explant is a flower pesticide, draws materials period: the monokaryon at flower pesticide is drawn materials mid-term;
Said MS medium comprises: MS+2, and 4-D2.0mg/L+6-BA1.5mg/L+NAA1.0mg/L comprises that also concentration is 6% sucrose;
This method also 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 201110046751 CN102144563B (en) | 2009-05-11 | 2009-05-11 | Method for inducing calluses of peony anther |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110046751 CN102144563B (en) | 2009-05-11 | 2009-05-11 | Method for inducing calluses of peony anther |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200910083744 Division CN101548647B (en) | 2009-05-11 | 2009-05-11 | Tree peony callus induction method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102144563A CN102144563A (en) | 2011-08-10 |
| CN102144563B true CN102144563B (en) | 2012-09-26 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN 201110046751 Expired - Fee Related CN102144563B (en) | 2009-05-11 | 2009-05-11 | Method for inducing calluses of peony anther |
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102577968B (en) * | 2012-03-06 | 2013-10-16 | 河南科技大学 | Adventitious bud induction method for tree peony |
| CN105331573A (en) * | 2015-09-25 | 2016-02-17 | 天津科技大学 | Method of preparing haploid cells of Althaea rosea |
| CN106613981A (en) * | 2016-12-23 | 2017-05-10 | 江苏农林职业技术学院 | Paeonia rockii primary culture callus induction method |
| CN108834892A (en) * | 2018-06-15 | 2018-11-20 | 浙江农林大学暨阳学院 | A kind of Chinese herbaceous peony callus induction method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1768576A (en) * | 2004-11-03 | 2006-05-10 | 中国科学院上海生命科学研究院 | Peony standardized quick-breeding method |
| CN1778168A (en) * | 2005-10-12 | 2006-05-31 | 北京林业大学 | Cultivation of peony germinal dislocation |
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- 2009-05-11 CN CN 201110046751 patent/CN102144563B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1768576A (en) * | 2004-11-03 | 2006-05-10 | 中国科学院上海生命科学研究院 | Peony standardized quick-breeding method |
| CN1778168A (en) * | 2005-10-12 | 2006-05-31 | 北京林业大学 | Cultivation of peony germinal dislocation |
Non-Patent Citations (4)
| Title |
|---|
| 李丽霞等.应用正交设计方法筛选牡丹(paeonia suffruticosa)愈伤诱导培养基的研究.《烟台大学学报(自然科学与工程版)》.2005,(第01期), * |
| 李艳敏等.牡丹离体培养与快速繁殖研究进展.《西南林学院学报》.2004,(第01期), * |
| 王军娥等.牡丹愈伤组织诱导与分化技术的优化研究.《西北农业学报》.2008,(第05期), * |
| 陈怡平等.紫斑牡丹休眠地下芽在组织培养条件下的发育研究.《西北植物学报》.2003,(第02期), * |
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| CN102144563A (en) | 2011-08-10 |
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