CN105359982A - Armeniaca sibirica tissue culture medium, preparation method and application thereof - Google Patents
Armeniaca sibirica tissue culture medium, preparation method and application thereof Download PDFInfo
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- CN105359982A CN105359982A CN201510932389.8A CN201510932389A CN105359982A CN 105359982 A CN105359982 A CN 105359982A CN 201510932389 A CN201510932389 A CN 201510932389A CN 105359982 A CN105359982 A CN 105359982A
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- ansu apricot
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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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- 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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Abstract
The invention relates to the field of plant tissue culture, in particular to an armeniaca sibirica tissue culture medium and application thereof. The rotting culture medium comprises a 1/2MS culture medium, agar with the concentration being 4-8 g/L, cane sugar with the concentration being 25-50 g/L, a plant growth regulating agent IBA with the concentration being 2-4 mg/L and 1500-3000-fold almond shell wood vinegar. A novel, simple and efficient armeniaca sibirica tissue culture seedling rooting method is built, armeniaca sibirica tissue culture seedlings good in rooting performance and high in rooting yield are obtained through quick and efficient induction, large-scale rooting quick induction of the armeniaca sibirica tissue culture seedlings is achieved under the condition of manual control, armeniaca sibirica tissue culture seedlings rooting induction efficiency is greatly improved, and the seedling culture process is greatly promoted.
Description
Technical field
The present invention relates to field of plant tissue culture, particularly a kind of ansu apricot tissue culture medium (TCM), its preparation method and application.
Background technology
Ansu apricot (Armeniacasibirica (L.) Lam) is rose family apricot platymiscium, is the woody grain and oil economic tree integrating ecological benefits, Social benefit and economic benefit.Ansu apricot producing region is classified as the emphasis nonwood forest trees of economic development, is the important guarantee of economic development, increasing peasant income.But the development of ansu apricot industry is subject to the obstruction of kind and cultivation management shortcoming.
The reform of ansu apricot planting system is being prepared in Pingquan County, Hebei province, and introducing good strains of seeds is necessary condition, but it is high to buy new varieties seedling cost, and does not carry out introduction and Experiment in locality, Chengde, there is certain risk; Although graft technology can realize the change of kind, but because grafting affinity, stock are on factors such as the impacts of grafting, whether the ansu apricot nursery stock after grafting can be consistent with original maternal character, also there is not more deep research, and become new varieties to be unpractical whole for existing wild apricot grafting yet.Therefore, the nursery stock excellent according to the planting environment selection traits of locality, producing region is needed to breed on a large scale for production.
In three northern areas of China, the invasion and attack often meeting cold current in early spring are produced in ansu apricot cultivation, cause the significantly underproduction, select cold-resistant, that high yield, late flowering kind meet this area planting environment.Each ansu apricot main producing region wild apricot type is enriched, wherein have cold-resistant, high yield, evening bloom, the kind of strong stress resistance, if by its culture and utility, greatly will promote the development of ansu apricot industry, but these wild resource quantity are few, and skewness, how obtaining a large amount of excellent nursery stock consistent with above-mentioned maternal plant merit is at short notice put the first road difficult problem in face of ansu apricot industry development is wanted.Plant clonal reproduction technology is for herein is provided a more reliable approach.Utilize plant clonal reproduction technology can obtain a large amount of nursery stock consistent with maternal plant proterties at short notice, and with low cost, not being subject to seasonal restrictions, is the Main Means of modern plant nursery.
If under condition of tissue culture, with the excellent ansu apricot individual plant of economical character for material, set up its high-efficient cloning breeding system, thus realize its factorial seedling growth, then can carry out throughout the year, greatly can improve the efficiency of ansu apricot breeding, nursery, shorten the progress of work.
CN103392597A discloses a kind of method for tissue culture of North American begonia, comprise Initial culture, squamous subculture, culture of rootage and acclimatization and transplants, shoot is given birth to then for explant with North American begonia, after aseptic inoculation cultivates a period of time on Initial culture base, squamous subculture is carried out by being seeded on subculture medium after the North American begonia seedling cutting on Initial culture base, be transferred to when tufted seedling grows to 2-3cm and root media carry out root induction and form seedling of taking root, seedling of taking root obtains regeneration plant through hardening domestication.The method of described tissue cultures has had more successful experience, but ansu apricot is more difficult takes root, and the method is but not too applicable to ansu apricot.
Liu little Lei etc. are published in the regeneration describing apricot in " tissue cultures of Siberia apricot " of Plant Physiology Communications the 44th volume the 3rd phase and cultivate, but what it adopted is callus regeneration seedling, what the report " Plant Regeneration from Cultured Protoplasts of Armeniaca vulgaris var. ansu " of Ma Fengwang adopted is protoplast regeneration seedling, not by parent collected specimens, through Initial culture and squamous subculture, the variation whether these in vitro seedlings itself there occurs on gene is not come to a conclusion, and does not meet the object of setting up a factory nursery system.
Summary of the invention
The present invention utilizes tissue culture technique, provides a kind of ansu apricot tissue culture medium (TCM), its preparation method and application, and the rooting rate of the ansu apricot tissue cultures come by Initial culture by this method has brought up to more than 95%.
For reaching above-mentioned purpose, the present invention adopts following technical scheme:
First aspect, the invention provides a kind of root media of ansu apricot tissue cultures, and described root media comprises 1/2MS medium, the agar of 4-8g/L, the sucrose of 25-50g/L, plant growth regulator IBA2-4mg/L and 15 Pericarppium Armeniacae Amarum pyroligneous liquor.
In the present invention, described ansu apricot shell pyroligneous liquor contributes to ansu apricot and takes root, and improves rooting rate, coordinates plant growth regulator, makes ansu apricot rooting rate to reach more than 95%, the long 1.68-1.98cm of average root, mean elements 4.2-5.78 bar.
Use 1/2MS medium, add the agar of 4-8g/L, 30g/L sucrose, the sucrose of 25-50g/L, plant growth regulator IBA2.0-4.0mg/L, pH value is adjusted to 5.8, at 121 DEG C after sterilizing 17min, add dilution 1500-2500 doubly, wild almond shell pyroligneous liquor after sterilizing, for subsequent use after it solidifies; To be seeded in the root media prepared through Initial culture, ansu apricot tissue culturing seedling after squamous subculture 2-3 generation, observe ansu apricot plantlet in vitro and to take root situation.Namely can be observed root after 7 days to grow, observe the situation of taking root of ansu apricot plantlet in vitro at any time.
Described agar can be 4g/L, 5g/L, 6g/L, 7g/L or 8g/L.
Described sucrose can be 25g/L, 26g/L, 28g/L, 30g/L, 32g/L, 33g/L, 35g/L, 36g/L, 38g/L, 40g/L, 42g/L, 45g/L, 46g/L, 48g/L or 50g/L.
Described plant growth regulator IBA can be 2mg/L, 2.5mg/L, 3mg/L, 3.5mg/L or 4mg/L.
Preferably, described root media is made up of the agar of 1/2MS medium, 5g/L, the sucrose of 30g/L, a plant growth regulator IBA2.0-3.0mg/L and 1500-3000 times Pericarppium Armeniacae Amarum pyroligneous liquor.Preferably, the pH value of described root media is 5.5-6, such as, can be 5.5,5.6,5.7,5.8,5.9 or 6, is preferably 5.8.
Second aspect, the invention provides a kind of preparation method of root media of ansu apricot tissue cultures, comprises the steps:
(1) prepare minimal medium of taking root, use 1/2MS medium, add 5.0g/L agar, 30g/L sucrose, plant growth regulator IBA2.5-3.0mg/L, pH value is adjusted to 5.8, and at 121 DEG C, sterilizing 17min is for subsequent use;
(2) adding after the pyroligneous liquor dilution 1500-3000 times of sterilizing of being made by ansu apricot shell takes root in minimal medium, for subsequent use after it solidifies, and both obtains ansu apricot plantlet in vitro root media;
(3) by through Initial culture, squamous subculture 2-3 generation after ansu apricot tissue culturing seedling be seeded in the root media (2) prepared.
The third aspect, the invention provides a kind of method of ansu apricot tissue cultures, comprises the steps: ansu apricot explant to carry out pretreatment, Initial culture, squamous subculture and culture of rootage;
Wherein, described culture of rootage adopts root media as described in relation to the first aspect.
Preferably, described ansu apricot explant is selected from stem section and/or the stem apex of ansu apricot.
The time of described Initial culture is 23-35 days, such as, can be 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days or 35 days, is preferably 26-32 days.
Preferably, the algebraically of described squamous subculture is 2-3 generation.
Preferably, the time of described squamous subculture is 15-20 days, such as, can be 15 days, 16 days, 17 days, 18 days, 19 days or 20 days.
In the present invention, the time of ansu apricot tissue cultures is short, and whole incubation only needs 2 months, shortens incubation time compared to additive method.
Preferably, described Initial culture base, subculture medium and root media all carried out sterilizing before cultivation ansu apricot.
Preferably, the temperature of described sterilizing is 110-125 DEG C, can be such as 110 DEG C, 111 DEG C, 112 DEG C, 113 DEG C, 115 DEG C, 116 DEG C, 118 DEG C, 120 DEG C, 121 DEG C, 122 DEG C, 123 DEG C or 125 DEG C, be preferably 115-123 DEG C, more preferably 121 DEG C.
Preferably, described sterilization time is 10-25min, such as, can be 10min, 12min, 13min, 15min, 16min, 18min, 19min, 20min, 21min, 22min, 23min or 25min, is preferably 15-20min, more preferably 17min.
Compared with prior art, tool of the present invention has the following advantages:
(1) rooting rate of the ansu apricot plantlet in vitro of the method cultivation of ansu apricot tissue cultures of the present invention can reach more than 95%, the long 1.68-1.98cm of average root, mean elements 4.2-5.78 bar, the rooting rate cultivating ansu apricot compared to other modes has exceeded one times, and incubation time is short, whole process only needs 2 months, shortens the time than what cultivate ansu apricot in other modes;
(2) ansu apricot in the method for ansu apricot tissue cultures of the present invention is all by parent collected specimens, can obtain a large amount of excellent nursery stock consistent with maternal plant merit, can bloom in cold-resistant, high yield, evening, strong stress resistance;
(3) the present invention establishes a kind of easy, efficient group of training ansu apricot plantlet in vitro and to take root new method, fast, efficiently can induce the ansu apricot plantlet in vitro plant obtaining taking root, rooting rate is high, can under manual control condition, realize the scale of ansu apricot plantlet in vitro to take root rapid induction, greatly improve ansu apricot plantlet in vitro root induction efficiency and breeding process;
(4) the present invention can provide basis for the factorial seedling growth of ansu apricot improved seeds, and application prospect is boundless, significant for development mountain area ansu apricot industry, economic development and increasing peasant income.
Embodiment
For further setting forth the technological means and effect thereof that the present invention takes, further illustrate technical scheme of the present invention below in conjunction with the preferred embodiments of the present invention, but the present invention is not confined in scope of embodiments.
Embodiment
(1) first prepare ansu apricot to take root minimal medium, use 1/2MS medium, add 5.0g/L agar, 30g/L sucrose, plant growth regulator IBA2.5-3.0mg/L, pH value is adjusted to 5.8, and at 121 DEG C, sterilizing 17min is for subsequent use;
(2) the pyroligneous liquor dilution 1500-2500 made by ansu apricot shell doubly, adds after sterilizing and takes root in minimal medium, for subsequent use after it solidifies, and both obtains ansu apricot plantlet in vitro root media;
(3) will be seeded in the root media (2) prepared on superclean bench through pretreatment, Initial culture, ansu apricot tissue culturing seedling after squamous subculture 2-3 generation, observe ansu apricot plantlet in vitro and to take root situation.
Namely can be observed root after general 7 days to grow, recorded the situation of taking root of ansu apricot plantlet in vitro at any time, after the phenomenon that occurs taking root 15 days, the rooting rate of ansu apricot plantlet in vitro can reach more than 95%, the long 1.68-1.98cm of average root, mean elements 4.2-5.78 bar.
In sum, the rooting rate of the ansu apricot plantlet in vitro of the method cultivation of ansu apricot tissue cultures of the present invention is high, the long 1.68-1.98cm of average root, mean elements 4.2-5.78 bar, and the rooting rate cultivating ansu apricot compared to other modes has exceeded one times; And the present invention establishes a kind of easy, efficient group of training ansu apricot plantlet in vitro takes root new method; growth cycle is short; only need just can cultivate for 2 months to take root; the time is shortened than additive method; efficiently can induce the ansu apricot plantlet in vitro plant obtaining taking root, rooting rate is high; under manual control condition, realize the scale of ansu apricot plantlet in vitro and to take root rapid induction, greatly improve ansu apricot plantlet in vitro root induction efficiency and breeding process.
Applicant states, the present invention illustrates method detailed of the present invention by above-described embodiment, but the present invention is not limited to above-mentioned method detailed, does not namely mean that the present invention must rely on above-mentioned method detailed and could implement.Person of ordinary skill in the field should understand, any improvement in the present invention, to equivalence replacement and the interpolation of auxiliary element, the concrete way choice etc. of each raw material of product of the present invention, all drops within protection scope of the present invention and open scope.
Claims (9)
1. a root media for ansu apricot tissue cultures, is characterized in that, described root media comprises 1/2MS medium, the agar of 4-8g/L, the sucrose of 25-50g/L, plant growth regulator IBA2-4mg/L and 1500-3000 times Pericarppium Armeniacae Amarum pyroligneous liquor.
2. method according to claim 1, is characterized in that, described root media is made up of the agar of 1/2MS medium, 5g/L, the sucrose of 30g/L, a plant growth regulator IBA2.0-3.0mg/L and 1500-3000 times Pericarppium Armeniacae Amarum pyroligneous liquor.
3. method according to claim 1 and 2, is characterized in that, the pH value of described root media is 5.5-6, is preferably 5.8.
4. a preparation method for the root media of ansu apricot tissue cultures, is characterized in that, comprises the steps:
(1) by 1/2MS medium, add agar by formula ratio, sucrose, plant growth regulator IBA, adjust ph, sterilizing is for subsequent use;
(2) adding after sterilizing after the pyroligneous liquor dilution of being made by ansu apricot shell takes root in minimal medium, for subsequent use after it solidifies, and both obtains the root media of ansu apricot tissue cultures.
5. a method for ansu apricot tissue cultures, is characterized in that, comprises the steps: ansu apricot explant to carry out pretreatment, Initial culture, squamous subculture and culture of rootage;
Wherein, described culture of rootage adopts the root media according to any one of claim 1-3.
6. method according to claim 5, is characterized in that, described ansu apricot explant is selected from stem section and/or the stem apex of ansu apricot.
7. the method according to claim 5 or 6, is characterized in that, the time of described Initial culture is 23-35 days, is preferably 26-32 days.
8. the method according to any one of claim 5-7, is characterized in that, the algebraically of described squamous subculture is 2-3 generation;
Preferably, the time of described squamous subculture is 15-20 days.
9. the method according to any one of claim 5-8, is characterized in that, uses after described Initial culture base, subculture medium and root media all need sterilizing;
Preferably, the temperature of described sterilizing is 110-125 DEG C, is preferably 115-123 DEG C, more preferably 121 DEG C;
Preferably, described sterilization time is 10-25min, is preferably 15-20min, more preferably 17min.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106718893A (en) * | 2016-12-06 | 2017-05-31 | 内蒙古农业大学 | The method that Siberia apricot somatic embryo occurs |
| CN106962194A (en) * | 2017-04-11 | 2017-07-21 | 乌鲁木齐天格尔农业科技开发有限公司 | The method for tissue culture of Chinese bush cherry |
| CN109287491A (en) * | 2018-11-29 | 2019-02-01 | 钟天路 | A kind of almond tree tissue culture and rapid propagation method |
| CN115885848A (en) * | 2022-11-16 | 2023-04-04 | 北京林业大学 | Method for inducing proliferation and rooting of cluster buds of wild apricot |
| CN117305217A (en) * | 2023-10-24 | 2023-12-29 | 临沂市农业科学院 | Optimized stem tip tissue culture medium, culture method and application |
-
2015
- 2015-12-15 CN CN201510932389.8A patent/CN105359982B/en not_active Expired - Fee Related
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| 施琳: "山杏壳木醋液有效化学成分及生物活性研究", 《西北农林科技大学硕士学位论文》 * |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106718893A (en) * | 2016-12-06 | 2017-05-31 | 内蒙古农业大学 | The method that Siberia apricot somatic embryo occurs |
| CN106718893B (en) * | 2016-12-06 | 2018-09-14 | 内蒙古农业大学 | The method that Siberia apricot somatic embryo occurs |
| CN106962194A (en) * | 2017-04-11 | 2017-07-21 | 乌鲁木齐天格尔农业科技开发有限公司 | The method for tissue culture of Chinese bush cherry |
| CN109287491A (en) * | 2018-11-29 | 2019-02-01 | 钟天路 | A kind of almond tree tissue culture and rapid propagation method |
| CN115885848A (en) * | 2022-11-16 | 2023-04-04 | 北京林业大学 | Method for inducing proliferation and rooting of cluster buds of wild apricot |
| CN115885848B (en) * | 2022-11-16 | 2023-09-19 | 北京林业大学 | Method for inducing proliferation and rooting of cluster buds of wild apricots |
| CN117305217A (en) * | 2023-10-24 | 2023-12-29 | 临沂市农业科学院 | Optimized stem tip tissue culture medium, culture method and application |
| CN117305217B (en) * | 2023-10-24 | 2024-08-06 | 临沂市农业科学院 | Optimized stem tip tissue culture medium, culture method and application |
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