CN107711499A - A kind of BA of different time points 6 processing suppresses the method that training seedling adventitious root occurs - Google Patents
A kind of BA of different time points 6 processing suppresses the method that training seedling adventitious root occurs Download PDFInfo
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- CN107711499A CN107711499A CN201710989922.3A CN201710989922A CN107711499A CN 107711499 A CN107711499 A CN 107711499A CN 201710989922 A CN201710989922 A CN 201710989922A CN 107711499 A CN107711499 A CN 107711499A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
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Abstract
The invention belongs to industrial crops planting technology field, discloses a kind of BA of different time points 6 processing and suppresses the method that training seedling adventitious root occurs, is transferred in 3d and 7d and suppresses to cultivate in root media；6 BA concentration for the treatment of 2mg/L, are cultivated to 20d；Apple rootstock M26 tissue-cultured seedling needs tissue-cultured seedling after breeding culture medium culture is expanded one month before handling, the apple rootstock M26 tissue-cultured seedling that selection height reaches 1.5cm takes out from expansion breeding culture medium, is transferred to root media.The present invention can determine the problem of CK suppresses the period that adventitious root occurs, and suppress adventitious root genesis mechanism for CK, to instructing actual production tool to be of great significance.Occur mainly be transferred to the inhibitory action for suppressing root media 3 7d, CK to adventitious root dependent on the generation for suppressing adventitious root primordia present invention discover that CK suppresses adventitious root.
Description
Technical field
The invention belongs to industrial crops planting technology field, more particularly to a kind of different time points 6-BA processing to suppress training seedling
The method that adventitious root occurs.
Background technology
Adventitious root is the root that plant is occurred by improper sequential and happening part is indefinite, occurred in cane, leaf or
The organs such as hypocotyl.The generation of adventitious root be mostly because the organ or tissue of plant come to harm, hormone or biotic etc.
Stimulate, and by cell division, differentiation and development be root restriction by root initial body, then developed and formed by root restriction.Group in this research
Training seedling is probably to receive injured and hormone signal, adventitious root to produce from stem section.According to the research of forefathers, adventitious root hair
Life can be divided into three phases：Root induction stage, adventitious root primordia formation stages and adventitious root produce the stage.6-BA is
The basic element of cell division of first man work synthesis, 6-BA poor mobilities in plant, its physiological function is in treatment site and attached
Closely play a role, be advantageous to improve the accuracy of experiment with 6-BA processing tissue-cultured seedling.Many researchs have shown that the basic element of cell division
Suppress the generation of adventitious root, but basic element of cell division inhibitory action from which of indefinite root development is not known in period.Herein
In research, determine that inhibitory action can be by reducing the cell in this stage from which of indefinite root development in period for the basic element of cell division
Divide cellulose content to promote adventitious root to produce, be advantageous to the vegetative propagation of dwarfing rootstock, cultivate the apple rootstock material easily taken root.
And existing technology is all concentrated on and promoted in adventitious root mechanism, the method for inducing adventitious root is relatively simple.
In summary, the problem of prior art is present be：Existing technology concentrate on promote adventitious root generation, method compared with
To be single.
The content of the invention
The problem of existing for prior art, it is indefinite to suppress training seedling the invention provides a kind of different time points 6-BA processing
The method that root occurs.
The present invention is achieved in that a kind of different time points 6-BA processing suppresses the method that training seedling adventitious root occurs, institute
State the method that different time points suppress the adventitious root generation of training seedling using artificial synthesized basic element of cell division 6-BA processing, apple rootstock
M26 tissue-cultured seedling is transferred in 3d and 7d to be suppressed to cultivate in root media；6-BA concentration for the treatment of 2mg/L, are cultivated to 20d.
Further, the apple rootstock M26 tissue-cultured seedling Initial cultures：In the mother of big Tanaka's growth selection stalwartness no disease and pests harm
Strain, annual shoot is taken March, place Greenhouse Water Culture, the clip tender shoots when the bud length on branch is to 1.5-2cm, removal is unfolded
Blade, by stem apex in running water undershoot 6-8h, bud is taken out and is placed in wide-mouth bottle, carry out disinfection inoculation in superclean bench；
Material carries out disinfection processing, first with 75% ethanol postincubation 2s, is subsequently placed in 0.1% mercuric chloride solution processing 7min, then with sterilizing
Distilled water flushing 4~5 times, bud is cleaned up.Tender shoots after sterilization is inoculated into MS+6-BA 0.6mg/L+IBA 0.1mg/L+
PVP。
Further, the apple rootstock M26 tissue-cultured seedling squamous subcultures, by M26 tissue-cultured seedling from primary in superclean bench
Media transfer is to subculture medium.
Further, tissue-cultured seedling is needed to expand breeding culture medium culture one month before the apple rootstock M26 tissue-cultured seedling processing
Afterwards, choose height and reach 1.5cm apple rootstock M26 tissue-cultured seedling and take out from expanding in breeding culture medium, be transferred to root media.
Further, the expansion breeding culture medium be MS culture mediums, by IBA 0.1mg/L, 6-BA 0.6mg/L, agar 8g/L with
Sucrose 30g/L is formed, pH 5.8.
Further, the root media is by 1/2MS culture mediums, IBA 1mg/L compositions, pH 5.8.
Further, root media is suppressed by 1/2MS culture mediums, IBA 1mg/L, 6-BA 2mg/L compositions, pH 5.8.
Further, the apple rootstock M26 tissue-cultured seedling culturing room illumination 16h/d, illumination 2000lux, temperature are 25 DEG C, are put down
Equal humidity is 70% -80%.
Another object of the present invention is to provide a kind of side for suppressing apple rootstock M26 tissue-cultured seedling adventitious root and occurring
Method, be advantageous to the dwarfing rootstock that seed selection is easily taken root, play an important roll to breeding for China's Dwarf Stocks For Apple Trees.
The present invention can determine the problem of CK suppresses the period that adventitious root occurs, and suppress adventitious root genesis mechanism for CK, short
Change stock mainly to be bred by vegetative manner, the inhibiting mechanism that adventitious root of the present invention occurs, be advantageous to seed selection appearance
The dwarfing rootstock easily taken root, to instructing actual production tool to be of great significance.Present invention discover that it is transferred to suppression life in 3d
Cultivated in root culture medium, suppress to contain 2mg/L6-BA in root media, there is no the generation of adventitious root after cultivating 20d；7d is transferred to
Suppress there is part adventitious root to produce after cultivating 20d in root media.Occur mainly turning present invention demonstrates that CK suppresses adventitious root
Move to and suppress root media 3-7d.
Brief description of the drawings
Fig. 1 is that different time points 6-BA processing provided in an embodiment of the present invention suppresses the method flow that training seedling adventitious root occurs
Figure.
Fig. 2 is that different time points 6-BA processing rooting rate provided in an embodiment of the present invention counts schematic diagram, variance analysis P<
0.05。
Fig. 3 is different time points 6-BA processing adventitious root number statistical schematic diagram provided in an embodiment of the present invention, and difference is divided
Analyse P<0.05.
Fig. 4 is different time points 6-BA processing adventitious roots length statistics schematic diagram provided in an embodiment of the present invention, and difference is divided
Analyse P<0.05.
Embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
Different time points 6-BA processing provided by the invention suppresses the method that apple rootstock M26 tissue-cultured seedling adventitious root occurs,
M26 is a kind of common dwarfing rootstock, has the rich fruit of premature labor, admittedly property and winter resistance it is stronger the advantages that, in recent years, M26 performances
Good cultivation prospect.
The application principle of the present invention is explained in detail below in conjunction with the accompanying drawings.
As shown in figure 1, different time points 6-BA processing provided in an embodiment of the present invention suppresses the side that training seedling adventitious root occurs
Method comprises the following steps：
S101：Apple rootstock M26 tissue-cultured seedling is illumination 16h/d, illumination expanding breeding culture medium culture one month, condition of culture
2000lux, temperature are 25 DEG C, and medial humidity is 70-80%；
S102：The apple rootstock M26 tissue-cultured seedling that selection height reaches 1.5cm takes out from expansion breeding culture medium, is transferred to and contains
1mg/L IBA root medias；
S103：3d (or 7d) is cultivated in root media it is transferred to IBA containing 1mg/L and 2mg/L6-BA and suppresses training of taking root
Support in base and cultivate to 20d.
In a preferred embodiment of the invention：Apple rootstock M26 tissue-cultured seedling Initial cultures：It is healthy and strong in big Tanaka's growth selection
The maternal plant of no disease and pests harm, annual shoot is taken March, place Greenhouse Water Culture, clip is tender when the bud length on branch is to 1.5-2cm
Blade is unfolded in bud, removal, by stem apex in running water undershoot 6-8h, bud is taken out and is placed in wide-mouth bottle, is entered in superclean bench
Row sterilization inoculation.Material need to carry out disinfection processing, first with 75% ethanol postincubation 2s, be subsequently placed in the processing of 0.1% mercuric chloride solution
7min, then rinsed 4~5 times with sterile purified water, bud is cleaned up.Tender shoots after sterilization is inoculated into MS+6-BA 0.6mg/L+
IBA 0.1mg/L+PVP (polyvinylpyrrolidone) 500mg/L culture medium (every liter of culture medium needs 30g sucrose, 8g agar).For
Cross pollution is avoided, each blake bottle is only inoculated with 1 bud, is cultivated in putting it between tissue cultures.
In a preferred embodiment of the invention：Expansion breeding culture medium is MS culture mediums, by IBA0.1mg/L, 6-BA0.6mg/L,
Agar 8g/L and sucrose 30g/L compositions, pH 5.8；Root media is made up of 1/2MS culture mediums, IBA 1mg/L, and pH is
5.8；Suppress root media by 1/2MS culture mediums, IBA 1mg/L, 6-BA2mg/L compositions, pH 5.8.
The tissue culture culture medium prescription of table 1
Fig. 2 different time points 6-BA processing rooting rate statistics, variance analysis P<0.05.
Rooting rate calculation formula is：(the tissue-cultured seedling number for the tissue-cultured seedling number/whole taken root) × 100.
Fig. 3 different time points 6-BA handles adventitious root number statistical, variance analysis P<0.05.
Fig. 4 different time points 6-BA processing adventitious root length statistics, variance analysis P<0.05.
The application principle of the present invention is further described with reference to specific embodiment.
Embodiment 1：The Initial culture of apple rootstock M26 tissue-cultured seedling, in the mother of big Tanaka's growth selection stalwartness no disease and pests harm
Strain, annual shoot is taken March, place Greenhouse Water Culture, the clip tender shoots when the bud length on branch is to 1.5-2cm, removal is unfolded
Blade, by stem apex in running water undershoot 6-8h, bud is taken out and is placed in wide-mouth bottle, carry out disinfection inoculation in superclean bench.
Material need to carry out disinfection processing, first with 75% ethanol postincubation 2s, be subsequently placed in 0.1% mercuric chloride solution processing 7min, then with going out
Bacterium distilled water flushing 4~5 times, bud is cleaned up.Tender shoots after sterilization is inoculated into MS+6-BA 0.6mg/L+IBA 0.1mg/L
+ PVP (polyvinylpyrrolidone) 500mg/L culture medium (every liter of culture medium needs 30g sucrose, 8g agar).It is dirty to avoid intersecting
Dye, each blake bottle is only inoculated with 1 bud, is cultivated in putting it between tissue cultures.
Embodiment 2：Apple rootstock M26 tissue culture plant inoculations connect in expanding breeding culture medium in each blake bottle of breeding culture medium is expanded
3 plants of apple rootstock M26 tissue-cultured seedling, cultivate one month, it is MS culture mediums to expand breeding culture medium formula, by IBA0.1mg/L, 6-
BA0.6mg/L, agar 8g/L and sucrose 30g/L are formed, pH 5.8；Apple rootstock M26 tissue-cultured seedling is expanding breeding culture medium culture one
After individual month, apple rootstock M26 tissue-cultured seedling bottom callus is cut in superclean bench, the apple rootstock M26 groups to expanding numerous acquisition
Train seedling and carry out plant division, the apple rootstock M26 tissue-cultured seedling that selection height reaches 1.5cm is transferred to root media.Root media
By 1/2MS culture mediums, IBA 1mg/L compositions, pH 5.8；Suppression is transferred in root media after culture 3d and 7d to take root
Cultivated in culture medium；Suppress root media by 1/2MS culture mediums, IBA 1mg/L, 6-BA2mg/L compositions, pH 5.8.
Embodiment 3：Culture (3d 6-BA in suppression root media are transferred to after 3d and 7d is cultivated in root media
With 7d 6-BA), cultivate to 20d and count rooting rate, rooting rate is as shown in Fig. 2 3d is transferred to the tissue culture suppressed in root media
Seedling rooting rate is that the tissue-cultured seedling rooting rate that zero, 7d is transferred in suppression root media is 48.2%.The adventitious root number of statistics
As shown in figure 3,3d, which is transferred to the tissue-cultured seedling suppressed in root media, does not have the generation of adventitious root, and 7d is transferred to suppression and taken root
Tissue-cultured seedling in culture medium generates a small amount of adventitious root.The adventitious root length of statistics is as shown in figure 4,7d is transferred to suppression takes root
Tissue-cultured seedling of the tissue-cultured seedling adventitious root length also than being cultivated in root media in culture medium is short.As a result show after 3d at 6-BA
The generation of reason complete inhibition adventitious root, 6-BA process parts suppress the generation of adventitious root after 7d.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
All any modification, equivalent and improvement made within refreshing and principle etc., should be included in the scope of the protection.
Claims (7)
1. a kind of different time points 6-BA processing suppresses the method that training seedling adventitious root occurs, it is characterised in that the different time
Point 6-BA processing suppresses the method apple rootstock M26 tissue-cultured seedling that training seedling adventitious root occurs and is transferred to suppression culture of rootage in 3d and 7d
Cultivated in base；6-BA concentration for the treatment of 2mg/L, are cultivated to 20d.
2. different time points 6-BA processing as claimed in claim 1 suppresses the method that training seedling adventitious root occurs, it is characterised in that
Initial culture is needed before the apple rootstock M26 tissue-cultured seedling processing：In the maternal plant of big Tanaka's growth selection stalwartness no disease and pests harm,
Annual shoot is taken March, places Greenhouse Water Culture, the clip tender shoots when the bud length on branch is to 1.5-2cm, leaf is unfolded in removal
Piece；Stem apex takes out in running water undershoot 6-8h, bud and is placed in wide-mouth bottle, and carry out disinfection inoculation in superclean bench.
3. different time points 6-BA processing as claimed in claim 2 suppresses the method that training seedling adventitious root occurs, it is characterised in that
The sterilization inoculation processing specifically includes：With 75% ethanol postincubation 2s, 0.1% mercuric chloride solution processing 7min is placed in, then with sterilizing
Distilled water flushing 4~5 times, bud is cleaned up；Tender shoots is inoculated into MS+6-BA 0.6mg/L+IBA 0.1mg/L+PVP after sterilization
500mg/L culture medium；Tissue-cultured seedling chooses the apple rootstock M26 that height reaches 1.5cm after breeding culture medium culture is expanded one month
Tissue-cultured seedling takes out from expansion breeding culture medium, is transferred to root media.
4. different time points 6-BA processing as claimed in claim 2 suppresses the method that training seedling adventitious root occurs, it is characterised in that
The expansion breeding culture medium is MS culture mediums, is made up of IBA 0.1mg/L, 6-BA 0.6mg/L, agar 8g/L and sucrose 30g/L,
PH is 5.8.
5. different time points 6-BA processing as claimed in claim 2 suppresses the method that training seedling adventitious root occurs, it is characterised in that
The root media is by 1/2MS culture mediums, IBA 1mg/L compositions, pH 5.8.
6. different time points 6-BA processing as claimed in claim 1 suppresses the method that training seedling adventitious root occurs, it is characterised in that
Suppress root media by 1/2MS culture mediums, IBA 1mg/L, 6-BA 2mg/L compositions, pH 5.8.
7. different time points 6-BA processing as claimed in claim 1 suppresses the method that training seedling adventitious root occurs, it is characterised in that
The apple rootstock M26 tissue-cultured seedling culturing room illumination 16h/d, illumination 2000lux, temperature are 25 DEG C, and humidity is 70% -80%.
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| CN201710989922.3A CN107711499A (en) | 2017-10-23 | 2017-10-23 | A kind of BA of different time points 6 processing suppresses the method that training seedling adventitious root occurs |
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| CN201710989922.3A CN107711499A (en) | 2017-10-23 | 2017-10-23 | A kind of BA of different time points 6 processing suppresses the method that training seedling adventitious root occurs |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115011610A (en) * | 2022-06-17 | 2022-09-06 | 西北农林科技大学 | Application of MdTCP17 and MdWOX11 in interaction regulation of MdLBD29 gene expression and adventitious root generation |
| CN116064572A (en) * | 2022-08-11 | 2023-05-05 | 西北农林科技大学 | MdWOX11 gene and protein for promoting adventitious root development and application thereof |
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| AU7323000A (en) * | 1999-08-26 | 2001-04-10 | Plant Research International B.V. | Conditional inhibition of vegetative propagation |
| EP2275564A1 (en) * | 2009-07-17 | 2011-01-19 | Freie Universität Berlin | Means and method for the production of transgenic plants that are resistant to clubroot |
| CN106134997A (en) * | 2016-07-06 | 2016-11-23 | 中国农业科学院郑州果树研究所 | The group training fast seedling-cultivating method of apple rootstock M26 |
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2017
- 2017-10-23 CN CN201710989922.3A patent/CN107711499A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU7323000A (en) * | 1999-08-26 | 2001-04-10 | Plant Research International B.V. | Conditional inhibition of vegetative propagation |
| EP2275564A1 (en) * | 2009-07-17 | 2011-01-19 | Freie Universität Berlin | Means and method for the production of transgenic plants that are resistant to clubroot |
| CN106134997A (en) * | 2016-07-06 | 2016-11-23 | 中国农业科学院郑州果树研究所 | The group training fast seedling-cultivating method of apple rootstock M26 |
Non-Patent Citations (4)
| Title |
|---|
| GEERT-JAN DE KLERK ET AL: "The formation of adventitious roots:new concepts,new possibilities", 《IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY. PLANT》 * |
| GEERT-JAN DE KLERK 等: "Timing of the phases in adventitious root formation in apple microcuttings", 《JOURNAL OF EXPERIMENTAL BOTANY》 * |
| 余亮: "苹果砧木M9和M26快繁体系的建立及移栽生理研究", 《中国优秀硕士学位论文数据库（硕士）》 * |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115011610A (en) * | 2022-06-17 | 2022-09-06 | 西北农林科技大学 | Application of MdTCP17 and MdWOX11 in interaction regulation of MdLBD29 gene expression and adventitious root generation |
| CN115011610B (en) * | 2022-06-17 | 2023-06-23 | 西北农林科技大学 | Application of MdTCP17 and MdWOX11 in interaction regulation and control of MdLBD29 gene expression and adventitious root generation |
| CN116064572A (en) * | 2022-08-11 | 2023-05-05 | 西北农林科技大学 | MdWOX11 gene and protein for promoting adventitious root development and application thereof |
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