CN108575749A - Atractylis lancea Aseptic seedling culture base and apply its Atractylis lancea Aseptic seedling culture technique - Google Patents
Atractylis lancea Aseptic seedling culture base and apply its Atractylis lancea Aseptic seedling culture technique Download PDFInfo
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- CN108575749A CN108575749A CN201810370488.5A CN201810370488A CN108575749A CN 108575749 A CN108575749 A CN 108575749A CN 201810370488 A CN201810370488 A CN 201810370488A CN 108575749 A CN108575749 A CN 108575749A
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- explant
- aseptic seedling
- atractylis lancea
- seedling 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
- A01H4/001—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
Abstract
The invention discloses a kind of Atractylis lancea Aseptic seedling culture base and apply its Atractylis lancea Aseptic seedling culture technique, Atractylis lancea Aseptic seedling culture base is comprising MS minimal mediums and adds 6 benayl aminopurines and methyl α-naphthyl acetate in MS minimal mediums respectively, wherein, 6 1.3~1.7mg of benayl aminopurine, 0.9~1.1mg of methyl α-naphthyl acetate are added in every liter of MS minimal medium.The Atractylis lancea Aseptic seedling culture base of the present invention improves the proliferation rate of explant, and average single bud proliferation quantity also increased, also shorten the seedling time of explant, improve economic benefit.
Description
Technical field
The present invention relates to a kind of Atractylis lancea Aseptic seedling culture base and apply its Atractylis lancea Aseptic seedling culture technique.
Background technology
Atractylis lancea is composite family rhizoma atractylodis number herbaceos perennial, and dry rhizome is famous traditional Chinese medicine, Genuine producing area
Rhizome main active for Region in Maoshan, Jiangsu Province, Atractylis lancea has Atisine chloride Atractydin, β-cineol etc..With drying damp and strengthening spleen, wind-dispelling
The effects that cold dispelling, cures mainly abdominal fullness and distention, diarrhea, oedema, arthralgia pain due to rheumatism, anemofrigid cold, incoordination between the spleen and the stomach, diarrhea, syndrome of dampness-heat, arthritis with fixed pain caused by dampness
With day disease, heat temperature, dysentery, when row diseases caused by external factors, the inferior disease of yellowish-white band.
In recent years, in the extensive application of the industries such as prepared slices of Chinese crude drugs factory, Chinese patent drug, market demand increasingly increases Atractylis lancea,
In planting process, Atractylis lancea seminal propagation germination percentage is low, and there are this quality deterioration, yield to reduce for root division, and disease aggravates
The problems such as.In existing Atractylis lancea group culturation rapid propagating technology, some scholars are set out by callus, induce Atractylis lancea budlet, but
In actual mechanical process, it there is a possibility that the easy browning of callus, increase the problems such as legacy variation.
Invention content
The technical problem to be solved by the present invention is to overcome the deficiencies of existing technologies, a kind of Atractylis lancea Aseptic seedling culture is provided
Base, it improves the proliferation rate of explant, and average single bud proliferation quantity also increased, when also having shortened the seedling of explant
Between, improve economic benefit.
In order to solve the above-mentioned technical problem, the technical scheme is that：A kind of Atractylis lancea Aseptic seedling culture base, it includes
MS minimal mediums and 6-benzyl aminopurine and methyl α-naphthyl acetate in MS minimal mediums are added respectively, wherein every liter of MS is basic
1.3~1.7mg of 6-benzyl aminopurine, 0.9~1.1mg of methyl α-naphthyl acetate are added in culture medium.
Further, MS minimal mediums add comprising MS culture mediums and respectively sucrose and agar in MS culture mediums,
In, 25~35g of sucrose is added in every liter of MS culture medium, the PH of agar 4.26~6.26g, MS minimal medium is 5.6~6.
The present invention also provides a kind of Atractylis lancea Aseptic seedling culture techniques, it applies Atractylis lancea Aseptic seedling culture base.
Further, include in processing step：
(1) explant samples：Take the bud on Atractylis lancea root as explant；
(2) explant sterilizes；
(3) explant is handled：Peel off the crust of explant volume surrounding browning；
(4) explant culture：By treated in step (3), explant is seeded on Atractylis lancea Aseptic seedling culture base, and
It is placed in incubator and carries out the Fiber differentiation of aseptic seedling.
Further, also include between step (1) and step (2)：Explant is rinsed well with tap water, with tap water
The process banister brush of flushing removes the outer layer bud skin of bud, then clean with pure water rinsing.
Further, in step (2), during carrying out disinfection to explant, first the explant rinsed well is positioned over
Mass concentration is to impregnate 1min in 75% alcoholic solution, and constantly concussion alcoholic solution is to increase connecing for explant and alcoholic solution
Contacting surface is accumulated, after explant is transferred in sterile water rinses 3~4 times, then with mass concentration be 0.1% mercuric chloride solution soaking disinfection 22
~28min, and mercuric chloride solution is constantly shaken, finally explant is transferred in sterile water again and is rinsed 3~5 times, that is, is completed to explant
The disinfection of body.
Further, in step (4), the condition of culture in incubator is：
Temperature：26.5~27.5 DEG C；
Light application time：12h；
Intensity of illumination is：3000Lx.
Further, fluorescent tube is cultivated by plant in incubator to complete the condition of culture of light application time and intensity of illumination.
After using above-mentioned technical proposal, the present invention is to be added to the 6-benzyl aminopurine of appropriate weight and the MS of methyl α-naphthyl acetate
Minimal medium improves bud appreciation rate as Atractylis lancea Aseptic seedling culture base, and the increment quantity of the bud of single explant is also big
It is big to increase, and increment bud growth is vigorous, the subculture for being conducive to the later stage is taken root, and has also been shortened the seedling time of explant, has been improved
Economic benefit；The Atractylis lancea Aseptic seedling culture technique of the present invention is using the radical bud of Atractylis lancea as explant, convenient material drawing, Atractylis lancea
Radical bud is half pack arrangement, the disinfecting time appropriate for extending disinfectant, the growing point inside bud can't browning lose growth energy
Pollution rate greatly reduces in power, improves conventional efficient.
Specific implementation mode
In order that the present invention can be more clearly and readily understood, below according to specific embodiment, to the present invention make into
One step is described in detail.
Embodiment one
A kind of Atractylis lancea Aseptic seedling culture base, it includes MS minimal mediums and adds respectively in MS minimal mediums
6-benzyl aminopurine and methyl α-naphthyl acetate, wherein 6-benzyl aminopurine 1.3mg, methyl α-naphthyl acetate are added in every liter of MS minimal medium
0.9mg。
MS minimal mediums are comprising MS culture mediums and add sucrose and agar in MS culture mediums respectively, wherein every liter
Sucrose 25g is added in MS culture mediums, the PH of agar 4.26g, MS minimal medium is 5.6.
A kind of Atractylis lancea Aseptic seedling culture technique, it applies Atractylis lancea Aseptic seedling culture base.
Include in processing step：
(1) explant samples：Take the bud on Atractylis lancea root as explant；
(2) explant sterilizes；
(3) explant is handled：Peel off the crust of explant volume surrounding browning；
(4) explant culture：By treated in step (3), explant is seeded on Atractylis lancea Aseptic seedling culture base, and
It is placed in incubator and carries out the Fiber differentiation of aseptic seedling.
Also include between step (1) and step (2)：Explant is rinsed well with tap water, what is rinsed with tap water
Process banister brush removes the outer layer bud skin of bud, then clean with pure water rinsing.
In step (2), during carrying out disinfection to explant, the explant rinsed well is first positioned over mass concentration
To impregnate 1min in 75% alcoholic solution, and constantly concussion alcoholic solution to be to increase the contact area of explant and alcoholic solution,
Explant is transferred in sterile water afterwards and is rinsed 3 times, then with mass concentration be 0.1% mercuric chloride solution soaking disinfection 22min, and constantly
Mercuric chloride solution is shaken, finally explant is transferred in sterile water again and is rinsed 3 times, that is, completes the disinfection to explant.
In step (4), the condition of culture in incubator is：
Temperature：26.5℃；
Light application time：12h；
Intensity of illumination is：3000Lx.
Fluorescent tube is cultivated to complete the condition of culture of light application time and intensity of illumination by plant in incubator.In the present embodiment
In, it is that T5 plants cultivate fluorescent tube that plant, which cultivates fluorescent tube, but not limited to this.
Embodiment two
A kind of Atractylis lancea Aseptic seedling culture base, it includes MS minimal mediums and adds respectively in MS minimal mediums
6-benzyl aminopurine and methyl α-naphthyl acetate, wherein 6-benzyl aminopurine 1.5mg, methyl α-naphthyl acetate are added in every liter of MS minimal medium
1.0mg。
MS minimal mediums are comprising MS culture mediums and add sucrose and agar in MS culture mediums respectively, wherein every liter
Sucrose 30g is added in MS culture mediums, the PH of agar 5.26g, MS minimal medium is 5.8.
A kind of Atractylis lancea Aseptic seedling culture technique, it applies Atractylis lancea Aseptic seedling culture base.
Include in processing step：
(1) explant samples：Take the bud on Atractylis lancea root as explant；
(2) explant sterilizes；
(3) explant is handled：Peel off the crust of explant volume surrounding browning；
(4) explant culture：By treated in step (3), explant is seeded on Atractylis lancea Aseptic seedling culture base, and
It is placed in incubator and carries out the Fiber differentiation of aseptic seedling.
Also include between step (1) and step (2)：Explant is rinsed well with tap water, what is rinsed with tap water
Process banister brush removes the outer layer bud skin of bud, then clean with pure water rinsing.
In step (2), during carrying out disinfection to explant, the explant rinsed well is first positioned over mass concentration
To impregnate 1min in 75% alcoholic solution, and constantly concussion alcoholic solution to be to increase the contact area of explant and alcoholic solution,
Explant is transferred in sterile water afterwards and is rinsed 4 times, then with mass concentration be 0.1% mercuric chloride solution soaking disinfection 25min, and constantly
Mercuric chloride solution is shaken, finally explant is transferred in sterile water again and is rinsed 5 times, that is, completes the disinfection to explant.
In step (4), the condition of culture in incubator is：
Temperature：27℃；
Light application time：12h；
Intensity of illumination is：3000Lx.
Fluorescent tube is cultivated to complete the condition of culture of light application time and intensity of illumination by plant in incubator.In the present embodiment
In, it is that T5 plants cultivate fluorescent tube that plant, which cultivates fluorescent tube, but not limited to this.
Embodiment three
A kind of Atractylis lancea Aseptic seedling culture base, it includes MS minimal mediums and adds respectively in MS minimal mediums
6-benzyl aminopurine and methyl α-naphthyl acetate, wherein 6-benzyl aminopurine 1.7mg, methyl α-naphthyl acetate are added in every liter of MS minimal medium
1.1mg。
MS minimal mediums are comprising MS culture mediums and add sucrose and agar in MS culture mediums respectively, wherein every liter
Sucrose 35g is added in MS culture mediums, the PH of agar 6.26g, MS minimal medium is 6.
A kind of Atractylis lancea Aseptic seedling culture technique, it applies Atractylis lancea Aseptic seedling culture base.
Include in processing step：
(1) explant samples：Take the bud on Atractylis lancea root as explant；
(2) explant sterilizes；
(3) explant is handled：Peel off the crust of explant volume surrounding browning；
(4) explant culture：By treated in step (3), explant is seeded on Atractylis lancea Aseptic seedling culture base, and
It is placed in incubator and carries out the Fiber differentiation of aseptic seedling.
Also include between step (1) and step (2)：Explant is rinsed well with tap water, what is rinsed with tap water
Process banister brush removes the outer layer bud skin of bud, then clean with pure water rinsing.
In step (2), during carrying out disinfection to explant, the explant rinsed well is first positioned over mass concentration
To impregnate 1min in 75% alcoholic solution, and constantly concussion alcoholic solution to be to increase the contact area of explant and alcoholic solution,
Explant is transferred in sterile water afterwards and is rinsed 4 times, then with mass concentration be 0.1% mercuric chloride solution soaking disinfection 28min, and constantly
Mercuric chloride solution is shaken, finally explant is transferred in sterile water again and is rinsed 5 times, that is, completes the disinfection to explant.
In step (4), the condition of culture in incubator is：
Temperature：27.5℃；
Light application time：12h；
Intensity of illumination is：3000Lx.
Fluorescent tube is cultivated to complete the condition of culture of light application time and intensity of illumination by plant in incubator.In the present embodiment
In, it is that T5 plants cultivate fluorescent tube that plant, which cultivates fluorescent tube, but not limited to this.
Such as table 1, in specific experiment, single use 6-benzyl aminopurine is that external source adds hormone first, when every liter of MS base
When the weighing less than 1.5mg of 6-benzyl aminopurine added in basal culture medium, bud is slow-growing, and the seedling time is longer, after seedling
Growing way is weaker；When the weight of the 6-benzyl aminopurine added in every liter of MS minimal medium reaches 1.5mg, bud growth fraction is very fast,
Bud appreciation rate also reaches highest, and bud growth of rising in value is vigorous；When the 6-benzyl aminopurine added in every liter of MS minimal medium
When weight is higher than 1.5mg, although there is vitrification phenomenon, is unfavorable for later stage culture of rootage also than very fast in bud growth after seedling.
But it was found that, when individually adding 6-benzyl aminopurine, the bud increasing quantity of single explant is seldom always.
Table 1：The 6-benzyl aminopurine of addition different weight trains Atractylis lancea radical bud aseptic seedling in every liter of MS minimal medium
Foster influence
Note：+ poor, ++ preferably, +++ it is vigorous
Such as table 2, in order to improve micropropagation efficiency, be added to suitable methyl α-naphthyl acetate, the methyl α-naphthyl acetate of low content to the proliferation of bud not
Obviously, when the weight of methyl α-naphthyl acetate in every liter of MS minimal medium reaches 1.0mg, bud proliferation rate highest, average single bud proliferation
Quantity is also most, and bud seedling speed is also most fast.When concentration is further added by, bud proliferation rate and proliferation quantity no longer rise, and bud at
Seedling speed starts slack-off, and blade starts yellowing.
Table 2：The 6-benzyl aminopurine of addition 1.5mg and the methyl α-naphthyl acetate of different weight are grey to thatch in every liter of MS minimal medium
The influence of art radical bud Aseptic seedling culture
Note：+ poor, ++ preferably, +++ it is vigorous
The present invention is to be added to the 6-benzyl aminopurine of appropriate weight and the MS minimal mediums of methyl α-naphthyl acetate as Atractylis lancea
Aseptic seedling culture base improves bud appreciation rate, and the increment quantity of the bud of single explant also greatly increases, and bud growth of rising in value is prosperous
It contains, the subculture for being conducive to the later stage takes root, also shortened the seedling time of explant, improved economic benefit；The thatch of the present invention is grey
Art Aseptic seedling culture technique is using the radical bud of Atractylis lancea as explant, convenient material drawing, and the radical bud of Atractylis lancea is half pack arrangement, appropriate
The disinfecting time for extending disinfectant, growing point inside bud can't browning lose growth ability, pollution rate greatly reduces,
Improve conventional efficient.
Particular embodiments described above, pair present invention solves the technical problem that, technical solution and advantageous effect carry out
It is further described, it should be understood that the above is only a specific embodiment of the present invention, is not limited to this
Invention, all within the spirits and principles of the present invention, any modification, equivalent substitution, improvement and etc. done should be included in this hair
Within bright protection domain.
Claims (8)
1. a kind of Atractylis lancea Aseptic seedling culture base, it includes MS minimal mediums, it is characterised in that：Also include to be added respectively in MS
6-benzyl aminopurine in minimal medium and methyl α-naphthyl acetate, wherein 6-benzyl aminopurine 1.3 is added in every liter of MS minimal medium
~1.7mg, 0.9~1.1mg of methyl α-naphthyl acetate.
2. Atractylis lancea Aseptic seedling culture base according to claim 1, it is characterised in that：MS minimal mediums are cultivated comprising MS
Base and sucrose and agar in MS culture mediums are added respectively, wherein 25~35g of sucrose, agar are added in every liter of MS culture medium
The PH of 4.26~6.26g, MS minimal medium is 5.6~6.
3. a kind of Atractylis lancea Aseptic seedling culture technique, it is characterised in that：Its application such as claim 1~2 any one of them thatch
Rhizoma atractylodis Aseptic seedling culture base.
4. Atractylis lancea Aseptic seedling culture technique according to claim 3, which is characterized in that include in processing step：
(1) explant samples：Take the bud on Atractylis lancea root as explant；
(2) explant sterilizes；
(3) explant is handled：Peel off the crust of explant volume surrounding browning；
(4) explant culture：By treated in step (3), explant is seeded on Atractylis lancea Aseptic seedling culture base, is placed in
The Fiber differentiation of aseptic seedling is carried out in incubator.
5. requiring the Atractylis lancea Aseptic seedling culture technique described in 4 according to complete clever power, which is characterized in that step (1) and step (2) it
Between also include：Explant is rinsed well with tap water, removes the outer layer bud of bud in the process banister brush rinsed with tap water
Skin is then clean with pure water rinsing.
6. Atractylis lancea Aseptic seedling culture technique according to claim 4, it is characterised in that：In step (2), to explant into
During row disinfection, the explant rinsed well is first positioned over mass concentration to impregnate 1min in 75% alcoholic solution, and
Constantly concussion alcoholic solution to increase the contact area of explant and alcoholic solution, after explant be transferred in sterile water rinse 3
~4 times, then with mass concentration be 0.1% mercuric chloride solution 22~28min of soaking disinfection, and mercuric chloride solution is constantly shaken, finally again
Explant is transferred in sterile water and is rinsed 3~5 times, that is, completes the disinfection to explant.
7. Atractylis lancea Aseptic seedling culture technique according to claim 4, it is characterised in that：In step (4), in incubator
Condition of culture is：
Temperature：26.5~27.5 DEG C；
Light application time：12h；
Intensity of illumination is：3000Lx.
8. Atractylis lancea Aseptic seedling culture technique according to claim 7, it is characterised in that：It is cultivated by plant in incubator
Fluorescent tube completes the condition of culture of light application time and intensity of illumination.
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| CN201810370488.5A CN108575749A (en) | 2018-04-24 | 2018-04-24 | Atractylis lancea Aseptic seedling culture base and apply its Atractylis lancea Aseptic seedling culture technique |
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| CN201810370488.5A CN108575749A (en) | 2018-04-24 | 2018-04-24 | Atractylis lancea Aseptic seedling culture base and apply its Atractylis lancea Aseptic seedling culture technique |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2021036472A1 (en) * | 2019-08-26 | 2021-03-04 | 南京农业大学 | Method for improving quality and transplanting survival rate of atractylodes lancea test-tube seedlings |
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| WO2021036472A1 (en) * | 2019-08-26 | 2021-03-04 | 南京农业大学 | Method for improving quality and transplanting survival rate of atractylodes lancea test-tube seedlings |
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Application publication date: 20180928 |