CN102172220B - 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 PDF

Info

Publication number
CN102172220B
CN102172220B CN 201110046908 CN201110046908A CN102172220B CN 102172220 B CN102172220 B CN 102172220B CN 201110046908 CN201110046908 CN 201110046908 CN 201110046908 A CN201110046908 A CN 201110046908A CN 102172220 B CN102172220 B CN 102172220B
Authority
CN
China
Prior art keywords
tree peony
callus
medium
blade
calluses
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Fee Related
Application number
CN 201110046908
Other languages
Chinese (zh)
Other versions
CN102172220A (en
Inventor
王雁
朱向涛
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Research Institute of Forestry of Chinese Academy of Forestry
Original Assignee
Research Institute of Forestry of Chinese Academy of Forestry
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Research Institute of Forestry of Chinese Academy of Forestry filed Critical Research Institute of Forestry of Chinese Academy of Forestry
Priority to CN 201110046908 priority Critical patent/CN102172220B/en
Publication of CN102172220A publication Critical patent/CN102172220A/en
Application granted granted Critical
Publication of CN102172220B publication Critical patent/CN102172220B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Landscapes

  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

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

Tree peony cauline leaf callus induction method
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 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 cauline leaf callus induction method of a kind of callus induction rate.
The objective of the invention is to realize through following technical scheme:
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, said tree peony explant comprises stem; Be the period of drawing materials: at the beginning of 4 months by the end of March spring, and the young tender stem section of getting the tender stem section of children or forming by seed sprouting;
The MS medium of stem comprises: MS+NAA0.05mg/L+6-BA2.0mg/L+2,4-D1.0mg/L.
Said 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, and the young tender blade of getting the tender blade of children or forming by seed sprouting;
The MS medium of blade comprises: MS+6-BA0.1mg/L+2,4-D0.5mg/L.
Technical scheme by the invention described above provides can be found out; 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, callus induction rate is high.
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 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%NaClO 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%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 young tender stem section that perhaps 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 young tender blade that perhaps 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 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 cauline leaf 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 comprises stem or blade;
Be the period of drawing materials of said stem: at the beginning of 4 months by the end of March spring, and the young tender stem section of getting the tender stem section of children or forming by seed sprouting;
The MS medium of stem comprises: MS+NAA0.05mg/L+6-BA2.0mg/L+2,4-D1.0mg/L;
Be the period of drawing materials of said blade: at the beginning of 4 months by the end of March spring, and the young tender blade of getting the tender blade of children or forming by seed sprouting;
The MS medium of blade comprises: MS+6-BA0.1mg/L+2,4-D0.5mg/L;
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.
CN 201110046908 2009-05-11 2009-05-11 Method for inducing calluses of stems and leaves of tree peony Expired - Fee Related CN102172220B (en)

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

Applications Claiming Priority (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

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
CN102172220A CN102172220A (en) 2011-09-07
CN102172220B true CN102172220B (en) 2012-09-26

Family

ID=44515615

Family Applications (1)

Application Number Title Priority Date Filing Date
CN 201110046908 Expired - Fee Related CN102172220B (en) 2009-05-11 2009-05-11 Method for inducing calluses of stems and leaves of tree peony

Country Status (1)

Country Link
CN (1) CN102172220B (en)

Families Citing this family (7)

* Cited by examiner, † Cited by third party
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
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
CN109602006B (en) * 2019-01-09 2022-07-01 山东贝世康生物科技有限公司 Culture purification method and application of active substance containing peony stem cells
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

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
倪德祥等.菊花器官培养的研究 Ⅰ.菊花叶组织培养中的器官发生.《西北植物学报》.1987,(第03期),
倪德祥等.菊花器官培养的研究 Ⅰ.菊花叶组织培养中的器官发生.《西北植物学报》.1987,(第03期), *
张玉芳.牡丹愈伤组织诱导的初步研究.《中国优秀硕士学位论文全文数据库(农业科技辑)》.2009,(第4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》.(第02期), *

Also Published As

Publication number Publication date
CN102172220A (en) 2011-09-07

Similar Documents

Publication Publication Date Title
CN101548647B (en) Tree peony callus induction method
CN102172220B (en) Method for inducing calluses of stems and leaves of tree peony
Dagla Plant tissue culture: Historical developments and applied aspects
Park et al. Improved shoot organogenesis of gloxinia ('Sinningia speciosa') using silver nitrate and putrescine treatment
Mohan et al. Plant regeneration in pigeonpea [Cajanus cajan (L.) Millsp.] by organogenesis
CN102550405A (en) Breeding method of poplar haploid
CN103444523B (en) Method for quickly introducing embryonic callus through anther to regenerate plant
CN104304030B (en) A kind of propagation method of oriental hybrid lily
Lin et al. In vitro flowering of green and albino Dendrocalamus latiflorus
CN102144563B (en) Method for inducing calluses of peony anther
CN103155862B (en) Sinocalamus latiflorus flower pesticide inductor embryo the method obtaining regeneration plant
CN106106178B (en) A kind of method for tissue culture of candy iris
Arora et al. Factors affecting somation embryogenesis in long term callus cultures of'safed musli'(Chlorophytum borivilianum), an endangered wonder herb
Li et al. Highly effective organogenesis and somatic embryogenesis of Clivia
CN108464242A (en) A kind of notable direct method for generation of Lilium tenuifolium somatic embryo for shortening induction time
ES2384876T3 (en) Highly efficient transformation and regeneration procedure of plant cells in suspension
CN109220809B (en) Koelreuteria paniculata somatic embryogenesis and plant regeneration culture method
CN101836589B (en) Method of rapid propagation of populus
Zhao et al. Rescue and in vitro culture of herbaceous peony immature embryos by organogenesis
Kumar Somatic embryogenesis and high frequency plantlet regeneration in callus cultures of Thevetia peruviana
CN102144564B (en) Method for inducing callus of peony petals
CN100442971C (en) Induction of peony embryoid
CN104719161B (en) Method for obtaining African daisy regeneration plant through inducing somatic embryo
CN103314849A (en) Wild tomato Solanum sitiens somatic embryo induction method and somatic embryo genetic transformation method
CN102511397A (en) Method for inducing populus calli and induction culture medium

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20120926

Termination date: 20170511

CF01 Termination of patent right due to non-payment of annual fee