CN105981651A - Subculture method of Cornus walteri Wanger. tissue culture seedlings - Google Patents
Subculture method of Cornus walteri Wanger. tissue culture seedlings Download PDFInfo
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- CN105981651A CN105981651A CN201510847105.5A CN201510847105A CN105981651A CN 105981651 A CN105981651 A CN 105981651A CN 201510847105 A CN201510847105 A CN 201510847105A CN 105981651 A CN105981651 A CN 105981651A
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- subculture
- cultured seedling
- tissue cultured
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- hair
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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
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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
Abstract
The invention discloses a subculture method of Cornus walteri Wanger. tissue culture seedlings. The subculture method includes: inoculating the bud tussocks, smaller than 1cm, of the sterile Cornus walteri Wanger. tissue culture seedlings satisfying subculture requirements into a proliferation culture medium, inoculating robust buds, not smaller than 1cm, of the tissue culture seedlings into an elongation culture medium, wherein the proliferation requirements of proliferation expanding propagation and high branches and robust seedlings are satisfied at the same. The subculture method has the advantages that the subculture method which is high in operability and combines proliferation and elongation growth is built by using different culture media, the average proliferation coefficient is above 10, average plant height is above 3.5cm, the tissue culture seedlings grow well, proliferation coefficient is stable after successive culture, and the subculture method is applicable to the large-scale production of the Cornus walteri Wanger. tissue culture seedlings and good in economic and social benefits.
Description
Technical field
The present invention relates to field of plant tissue culture, the subculture method of a kind of mao of tissue cultured seedling.
Background technology
Hair (Cornus walteri Wanger.) belongs to Fructus Corni Como and belongs to, and has another name called bicycle beam wood.Deciduous tree, up to 8-15m.
Liaoning is played in northeast, south to Hunan, southwest to Yunnan, Guizhou, east from Jiangsu, Zhejiang, west to Gansu, Qinghai all have distribution,
Wherein Shandong, Henan, Shaanxi, Shanxi distribution are more concentrated.Hair well developed root system, adaptive faculty is strong, requires soil not tight,
All suitable growth on the deserted mountain barren hill of China's most area.Hair is not only excellent commerical tree species, is also excellent simultaneously
Urban afforestation and water and soil conservation seeds, the good seeds of the biodiesel that develops forestry especially.Comospore meat and kernel all containing oils and fats,
Fruit oil content 31.8%-41.3%, its oil both edibles, can particularly can be made into biodiesel raw material as the raw material of industry again
Oil.
Due to the development and utilization being worth the fruit oils and fats of hair, development hair seeds also need to a large amount of high-quality clone kind at present
Seedling, and be provided that on market that the seedling of afforestation is also nowhere near, and quality is irregular differs.Cross unhairing propagation method with seed
Nursery is main, but the offspring of seminal propagation easily loses original merit, and seedling forestation typically takes 6-8 and starts knot
Really, yield height differs；The grafting general 2-3 that afforests starts result, but relatively costly and lack excellent strain scion；Cuttage is numerous
Growing the most relatively difficult, rooting rate is low, the most all cannot meet the demand of production.
In modern seedling-raising technique, plant tissue culture fast breeding technique can be bred by rapid, high volume at short notice, and it is consistent to obtain genotype
Good seed, it is adaptable to the factorial praluction of hair seeds nursery stock.But the research in terms of hair tissue culture rarely has at present
Report, is only limitted to initial culture, and still suffers from many problems.Therefore a kind of applicable hair tissue cultured seedling successive transfer culture is needed especially
Method, to solve the technical problem of above-mentioned existing existence.
Summary of the invention
It is an object of the invention to solve that hair tissue cultured seedling successive transfer culture growth coefficient is low, elongation growth slowly without problems such as branches of tall trees,
Optimization culture based formulas, it is provided that the subculture method of the high hair tissue cultured seedling with elongation growth of a kind of growth coefficient.
The subculture method of a kind of mao of tissue cultured seedling of the present invention, is achieved through the following technical solutions, comprises the following steps:
(1) enrichment culture: be up to the aseptic tissue cultured seedling of hair of subculture requirement, on superclean bench, the bud clump of ＜ 1cm cut
Being divided into little Cong to be inoculated in proliferated culture medium, condition of culture is 22-28 DEG C, intensity of illumination 1800-2500lx, illumination every day 12-16h,
Multiple Buds is formed after 1-2 subculture cycle；
(2) elongation cultivate: be up to the aseptic tissue cultured seedling of hair of subculture requirement, on superclean bench by tissue cultured seedling >=1cm
Strong bud branch be cut into >=0.5cm and at least containing the stem section of a pair bud, be inoculated in elongation medium, condition of culture is 22-28 DEG C,
Intensity of illumination 1800-2500lx, illumination every day 12-16h, available branches of tall trees strong sprout after 2-3 subculture cycle.
Above-described proliferation culture medium formula is: DKW+6-BA 0.1-1.0mg/L+IBA0.1-0.5mg/L+ sucrose 30g/L+
Agar powder 6.5g/L, pH are adjusted to 5.8-6.5.
Above-described elongation medium formula is: DKW+6-BA 0.2-1.0mg/L+NAA0.05-0.5mg/L+GA30.5-1.0
Mg/L+ sucrose 30g/L+ agar powder 6.5g/L, pH is adjusted to 5.8-6.5.
The subculture cycle of above-described hair subculture tissue cultured seedling is 20d.
The average proliferation coefficient of above-described hair subculture tissue cultured seedling is more than 10, and the hair tissue cultured seedling carrying out elongation cultivation is average
Every strain is containing more than 1.36 branches of tall trees (branch of plant height ＞ 2.5cm), and average plant height is more than 3.5cm.
The invention have the benefit that the present invention passes through coordinate plant growth regulator type and concentration, study it and hair group is trained
The impact of Seedling subculture multiplication, filters out corresponding culture medium prescription, provides equilibrium for hair tissue cultured seedling difference purpose successive transfer culture and closes
The nutritional need of reason, proliferated culture medium can realize the differentiation expanding propagation of bud, improve growth coefficient；Elongation medium can be effective
Promotion internode elongation, it is possible to quickly form branches of tall trees；Bud clump and the strong bud branch difference of >=1cm to hair tissue cultured seedling ＜ 1cm
Use the culture medium of correspondence, meet the propagation requirement in propagation expanding propagation and branches of tall trees strong sprouts；Establish a set of workable, propagation stretch
Long growth combines the successive transfer culture technology of hair tissue cultured seedling, and growth coefficient reaches more than 10, average more than plant height 3.5cm, and group
Seedlings cultivating grows fine, stable through too much culture growth coefficient, can be applicable in mao tissue cultured seedling large-scale production, has good
Economic benefit and social benefit.
Detailed description of the invention
Following example are to be discussed further the present invention, are not limitations of the present invention.
Embodiment 1:
The subculture method of a kind of mao of tissue cultured seedling of the present invention, comprises the following steps:
(1) enrichment culture: be up to the aseptic tissue cultured seedling of hair of subculture requirement, on superclean bench, the bud clump of ＜ 1cm cut
It is divided into little Cong and is inoculated in the proliferated culture medium of DKW+6-BA 0.25mg/L+IBA0.1mg/L+ sucrose 30g/L+ agar powder 6.5g/L
In, condition of culture is 25 DEG C, intensity of illumination 1800lx, illumination every day 14h, grows thickly through 1 subculture multiplication week after date formation
Bud, average proliferation coefficient 10.27；
(2) elongation cultivate: be up to the aseptic tissue cultured seedling of hair of subculture requirement, on superclean bench by tissue cultured seedling >=1cm
Strong bud branch be cut into >=0.5cm and at least containing the stem section of a pair bud, be inoculated in DKW+6-BA0.25mg/L
+NAA0.3mg/L+GA3In the elongation medium of 0.5mg/L+ sucrose 30g/L+ agar powder 6.5g/L, condition of culture is 25 DEG C,
Intensity of illumination 2500lx, illumination every day 15h, 2 subculture multiplication week after date can get branches of tall trees strong sprouts, average every strain is containing 1.36
Branches of tall trees (branch of plant height ＞ 2.5cm), average plant height is 3.51cm, growth coefficient average out to 10.18.
Embodiment 2:
The subculture method of a kind of mao of tissue cultured seedling of the present invention, comprises the following steps:
(1) enrichment culture: be up to the aseptic tissue cultured seedling of hair of subculture requirement, on superclean bench, the bud clump of ＜ 1cm cut
It is divided in the proliferated culture medium that little Cong is inoculated in DKW+6-BA1mg/L+IBA0.5mg/L+ sucrose 30g/L+ agar powder 6.5g/L,
Condition of culture is 25 DEG C, intensity of illumination 2000lx, illumination every day 14h, through 2 subculture multiplication week after date formation Multiple Buds,
Average proliferation coefficient 12.33；
(2) elongation cultivate: be up to the aseptic tissue cultured seedling of hair of subculture requirement, on superclean bench by tissue cultured seedling >=1cm
Strong bud branch be cut into >=0.5cm and at least containing the stem section of a pair bud, be inoculated in DKW+6-BA0.5mg/L
+NAA0.4mg/L+GA3In the elongation medium of 0.8mg/L+ sucrose 30g/L+ agar powder 6.5g/L, condition of culture is 25 DEG C,
Intensity of illumination 2500lx, illumination every day 15h, 2 subculture multiplication week after date can get branches of tall trees strong sprouts, average every strain is containing 1.77
Branches of tall trees (branch of plant height ＞ 2.5cm), average plant height is 3.83cm, growth coefficient average out to 12.35.
Embodiment 3:
The subculture method of a kind of mao of tissue cultured seedling of the present invention, comprises the following steps:
(1) enrichment culture: be up to the aseptic tissue cultured seedling of hair of subculture requirement, on superclean bench, the bud clump of ＜ 1cm cut
It is divided into little Cong or simple bud is inoculated in the propagation of DKW+6-BA 0.5mg/L+IBA0.8mg/L+ sucrose 30g/L+ agar powder 6.5g/L
In culture medium, condition of culture is 25 DEG C, intensity of illumination 1800lx, illumination every day 14h, through 2 subculture multiplication week after date shape
Become Multiple Buds, average proliferation coefficient 14.18；
(2) elongation cultivate: be up to the aseptic tissue cultured seedling of hair of subculture requirement, on superclean bench by tissue cultured seedling >=1cm
Strong bud branch be cut into >=0.5cm and at least containing the stem section of a pair bud, be inoculated in DKW+6-BA1mg/L+NAA0.5
mg/L+GA3In the elongation medium of 1mg/L+ sucrose 30g/L+ agar powder 6.5g/L, condition of culture is 25 DEG C, intensity of illumination
2500lx, illumination every day 15h, 3 subculture multiplication week after date can get branches of tall trees strong sprouts, average every strain is containing 2.04 branches of tall trees (plant heights
The branch of ＞ 2.5cm), average plant height is 4.07cm, growth coefficient average out to 13.38.
Above example is only to be described the preferred embodiment of the present invention, is not defined the scope of the present invention,
Technical scheme is carried out various deformation and improvement, all under other any spirit without departing from the present invention and principle
Should fall in the protection domain that claims of the present invention determines.
Claims (6)
1. the subculture method of a hair tissue cultured seedling, it is characterised in that comprise the following steps:
(1) enrichment culture: be up to the aseptic tissue cultured seedling of hair of subculture requirement, on superclean bench, the bud clump of ＜ 1cm cut
Being divided into little Cong to be inoculated in proliferated culture medium, condition of culture is 22～28 DEG C, intensity of illumination 1800～2500lx, illumination every day 12～
16h, forms Multiple Buds after 1-2 subculture cycle；
(2) elongation cultivate: be up to the aseptic tissue cultured seedling of hair of subculture requirement, on superclean bench by tissue cultured seedling >=1cm
Strong bud branch be cut into >=0.5cm and at least containing the stem section of a pair bud, be inoculated in elongation medium, condition of culture is
22～28 DEG C, intensity of illumination 1800～2500lx, illumination every day 12～16h, available branches of tall trees strong sprout after 2-3 subculture cycle.
The subculture method of the most according to claim 1 mao of tissue cultured seedling, it is characterised in that: institute in step (1) (2)
The subculture cycle stated is 20d.
The subculture method of the most according to claim 1 mao of tissue cultured seedling, it is characterised in that: described in step (1)
Proliferation culture medium formula is: DKW+6-BA 0.1～1.0mg/L+IBA0.1～0.5mg/L+ sucrose 30g/L+ agar powder 6.5g/L,
PH is adjusted to 5.8～6.5.
The subculture method of the most according to claim 1 mao of tissue cultured seedling, it is characterised in that: increase described in step (1)
Growing the hair tissue cultured seedling average proliferation coefficient that culture medium obtains is more than 10.
The subculture method of the most according to claim 1 mao of tissue cultured seedling, it is characterised in that: described in step (2)
Elongation medium formula is: DKW+6-BA 0.2～1.0mg/L+NAA0.05～0.5mg/L+GA30.5～1.0mg/L+ sucrose
30g/L+ agar powder 6.5g/L, pH are adjusted to 5.8～6.5.
The subculture method of the most according to claim 1 mao of tissue cultured seedling, it is characterised in that: step is stretched described in (2)
The hair tissue cultured seedling average proliferation coefficient that long culture medium obtains is more than 10, and average every strain is containing 1.36 branches of tall trees (plant height ＞ 2.5cm
Branch) more than, average plant height is more than 3.5cm.
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Cited By (2)
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| CN108575762A (en) * | 2018-07-18 | 2018-09-28 | 苏州枫彩生态科技集团有限公司 | A method of improving red branch wood tissue-cultured seedling proliferation rate |
| CN112715368A (en) * | 2021-02-04 | 2021-04-30 | 山西省林业和草原科学研究院 | Cornus walteri stem induced plant regeneration culture medium and tissue culture and rapid propagation method thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108575762A (en) * | 2018-07-18 | 2018-09-28 | 苏州枫彩生态科技集团有限公司 | A method of improving red branch wood tissue-cultured seedling proliferation rate |
| CN112715368A (en) * | 2021-02-04 | 2021-04-30 | 山西省林业和草原科学研究院 | Cornus walteri stem induced plant regeneration culture medium and tissue culture and rapid propagation method thereof |
| CN112715368B (en) * | 2021-02-04 | 2022-06-28 | 山西省林业和草原科学研究院 | Cornus walteri stem induced plant regeneration culture medium and tissue culture and rapid propagation method thereof |
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