CN109042337A - A method of utilizing potato leaf bud callus fast seedling growing - Google Patents
A method of utilizing potato leaf bud callus fast seedling growing Download PDFInfo
- Publication number
- CN109042337A CN109042337A CN201811155517.2A CN201811155517A CN109042337A CN 109042337 A CN109042337 A CN 109042337A CN 201811155517 A CN201811155517 A CN 201811155517A CN 109042337 A CN109042337 A CN 109042337A
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- China
- Prior art keywords
- culture medium
- culture
- leaf bud
- illumination
- potato
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Classifications
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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
-
- 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 method using potato leaf bud callus fast seedling growing, include the following steps: to prepare culture medium 1 and culture medium 2；Potato leaf bud is taken, washes, is impregnated with ethanol water, is taken out, then impregnated with liquor natrii hypochloritis, is taken out, with aseptic water washing, drying；Leaf bud is uniformly placed on culture medium 1, illumination, culture obtains irregular shape callus, and replacement culture medium 1 is cultivated, and blade generates filament shape root, it with scalpel cut-off blade, is laid in culture medium 1, illumination, cultivates, it is transferred to culture medium 2, illumination again, culture obtains a large amount of potato seedlings.Method seedling raise period of the invention is short, reduces the process of repeatedly replacement culture medium, the 300ml volume culture bottle containing 50ml culture medium, which can obtain 60-80 plants of potato seedlings, can be quickly obtained a large amount of seedling.
Description
Technical field
The invention belongs to molecular biology and agricultural technology field, and in particular to a kind of quick using potato leaf bud callus
The method of nursery.
Background technique
Potato category Solanaceae Solanum, annual herb plant, the big nutritional ingredient of yield are high.China be five big staple foods it
One, it is the third-largest important cereal crops in the whole world, therefore the fast seedling growing technology of potato is most important.Existing seedling-raising technique
Mainly there are seed seedling-raising and potato seed nursery, but both of which has what temperature sensitive, growing-seedling period was grown, cannot achieve
Cultivation seedling rapidly and efficiently.It is quick to potato because of a kind of method using potato leaf bud callus fast seedling growing of the invention
Nursery is particularly significant.
Summary of the invention
It is a kind of using potato the purpose of the present invention is overcoming the deficiencies of the prior art and provideLeaf budCallus fast seedling growing
Method.
Technical solution of the present invention is summarized as follows:
A method of using potato leaf bud callus fast seedling growing, include the following steps:
(1) add distilled water to 1L in 4.75gMS culture medium, 30g sucrose and 7g agar, mixing sterilizes, is cooled to 50 DEG C,
1mg methyl α-naphthyl acetate and 1mg6- benzyl aminoadenine is added, obtains culture medium 1；By 4.75gMS culture medium, 30g sucrose and 7g agar
Add distilled water to 1L, mixes, sterilizing is cooled to 50 DEG C, and 0.5mg methyl α-naphthyl acetate and 0.25mg6- benzyl aminoadenine is added, obtains
Culture medium 2；
(2) potato leaf bud is taken, it is water-soluble for 75% ethyl alcohol with volumetric concentration in sterile with distilled water flushing 3-4 times
Liquid impregnates 30-35s, takes out, then impregnates 8-15min with the liquor natrii hypochloritis that distilled water dilutes 1 times, takes out, is rushed with sterile water
It washes 2-3 times, is dried up in sterile；
(3) leaf bud that step (2) obtain uniformly is swung in the culture bottle equipped with culture medium 1, illumination, 23-27 DEG C of culture
Obtain irregular shape callus, replacement culture medium 1 is cultivated 17-26 days, blade generation filament shape root, with scalpel cut-off blade at
Within the scope of 1cm × 1cm, be laid in culture medium 1, illumination, 23-27 DEG C culture 12-18 days, then be transferred to culture medium 2, illumination, 23-
27 DEG C of cultures, obtain potato seedling.
Advantages of the present invention: method seedling raise period of the invention is short, reduces the process of repeatedly replacement culture medium, (contains
The 300ml volume culture bottle of 50ml culture medium can obtain 60-80 plants of potato seedlings) a large amount of seedling can be quickly obtained.
Specific embodiment
Liquor natrii hypochloritis is commercially available, and in terms of effective chlorine 10%.
The MS culture medium of each embodiment is free from sucrose.
The present invention is further illustrated combined with specific embodiments below.
Embodiment 1
A method of using potato leaf bud callus fast seedling growing, include the following steps:
(1) add distilled water to 1L in 4.75gMS culture medium, 30g sucrose and 7g agar, mixing sterilizes, is cooled to 50 DEG C,
1mg methyl α-naphthyl acetate and 1mg6- benzyl aminoadenine is added, obtains culture medium 1；By 4.75gMS culture medium, 30g sucrose and 7g agar
Add distilled water to 1L, mixes, sterilizing is cooled to 50 DEG C, and 0.5mg methyl α-naphthyl acetate and 0.25mg6- benzyl aminoadenine is added, obtains
Culture medium 2；
(2) potato leaf bud is taken, with distilled water flushing 4 times, it is 75% ethanol water that volumetric concentration is used in sterile
33s is impregnated, is taken out, then impregnates 10min with the liquor natrii hypochloritis that distilled water dilutes 1 times, is taken out, with aseptic water washing 3 times, in
It is dried up in sterile；
(3) leaf bud that step (2) obtain uniformly is swung in the culture bottle equipped with culture medium 1, illumination, 25 DEG C of cultures obtain
Irregular shape callus, replacement culture medium 1 are cultivated 20 days, and blade generates filament shape root, with scalpel cut-off blade at 1cm × 1cm
In range, 4 are taken to be laid in culture medium 1, illumination, 25 DEG C are cultivated 15 days, then are transferred to culture medium 2 (50ml culture medium 2 is installed in
In the culture bottle of 300ml volume), illumination, 25 DEG C of cultures (12 days) obtain (79 plants) a large amount of potato seedlings.
Embodiment 2
A method of using potato leaf bud callus fast seedling growing, include the following steps:
(1) with 1 step of embodiment (1)；
(2) potato leaf bud is taken, with distilled water flushing 3 times, it is 75% ethanol water that volumetric concentration is used in sterile
30s is impregnated, is taken out, then impregnates 8min with the liquor natrii hypochloritis that distilled water dilutes 1 times, is taken out, with aseptic water washing 2 times, in
It is dried up in sterile；
(3) leaf bud that step (2) obtain uniformly is swung in the culture bottle equipped with culture medium 1, illumination, 23 DEG C of cultures obtain
Irregular shape callus, replacement culture medium 1 are cultivated 26 days, and blade generates filament shape root, with scalpel cut-off blade at 1cm × 1cm
In range, 4 are taken to be laid in culture medium 1, illumination, 23 DEG C are cultivated 18 days, then are transferred to culture medium 2 (50ml culture medium 2 is installed in
In the culture bottle of 300ml volume), illumination, 23 DEG C of cultures (18 days) obtain (68 plants) a large amount of potato seedlings.
Embodiment 3
A method of using potato leaf bud callus fast seedling growing, include the following steps:
(1) with 1 step of embodiment (1)；
(2) potato leaf bud is taken, with distilled water flushing 4 times, it is 75% ethanol water that volumetric concentration is used in sterile
35s is impregnated, is taken out, then impregnates 15min with the liquor natrii hypochloritis that distilled water dilutes 1 times, is taken out, with aseptic water washing 3 times, in
It is dried up in sterile；
(3) leaf bud that step (2) obtain uniformly is swung in the culture bottle equipped with culture medium 1, illumination, 27 DEG C of cultures obtain
Irregular shape callus, replacement culture medium 1 are cultivated 17 days, and blade generates filament shape root, with scalpel cut-off blade at 1cm × 1cm
In range, 4 are taken to be laid in culture medium 1, illumination, 27 DEG C are cultivated 12 days, then are transferred to culture medium 2 (50ml culture medium 2 is installed in
In the culture bottle of 300ml volume), illumination, 27 DEG C of cultures (12 days) obtain a large amount of (74 plants) potato seedlings.
The above content is being further elaborated with for the invention, is not intended to limit the invention creation, all in this hair
Made any modifications, equivalent replacements, and improvements etc., should be included in the invention within the spirit and principle of bright creation
Within protection scope.
Claims (1)
1. a kind of method using potato leaf bud callus fast seedling growing, it is characterized in that including the following steps:
(1) add distilled water to 1L in 4.75gMS culture medium, 30g sucrose and 7g agar, mix, sterilizing is cooled to 50 DEG C, is added
1mg methyl α-naphthyl acetate and 1mg6- benzyl aminoadenine, obtain culture medium 1；4.75gMS culture medium, 30g sucrose and 7g agar are added into steaming
Distilled water mixes, sterilizing is cooled to 50 DEG C, and 0.5mg methyl α-naphthyl acetate and 0.25mg6- benzyl aminoadenine is added, is cultivated to 1L
Base 2；
(2) potato leaf bud is taken, with distilled water flushing 3-4 times, it is the leaching of 75% ethanol water that volumetric concentration is used in sterile
30-35s is steeped, is taken out, then impregnates 8-15min with the liquor natrii hypochloritis that distilled water dilutes 1 times, is taken out, with aseptic water washing 2-3
It is secondary, it is dried up in sterile；
(3) leaf bud that step (2) obtain uniformly is swung in the culture bottle equipped with culture medium 1, illumination, 23-27 DEG C of culture obtains
Irregular shape callus, replacement culture medium 1 cultivate 17-26 days, blade generation filament shape root, with scalpel cut-off blade at 1cm ×
Within the scope of 1cm, be laid in culture medium 1, illumination, 23-27 DEG C culture 12-18 days, then be transferred to culture medium 2, illumination, 23-27 DEG C
Culture, obtains a large amount of potato seedlings.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811155517.2A CN109042337A (en) | 2018-09-30 | 2018-09-30 | A method of utilizing potato leaf bud callus fast seedling growing |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811155517.2A CN109042337A (en) | 2018-09-30 | 2018-09-30 | A method of utilizing potato leaf bud callus fast seedling growing |
Publications (1)
| Publication Number | Publication Date |
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| CN109042337A true CN109042337A (en) | 2018-12-21 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811155517.2A Pending CN109042337A (en) | 2018-09-30 | 2018-09-30 | A method of utilizing potato leaf bud callus fast seedling growing |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH04287623A (en) * | 1991-03-15 | 1992-10-13 | Mitsui Petrochem Ind Ltd | Method for proliferating bulbs by plant tissue culture |
| CN101790935A (en) * | 2010-03-31 | 2010-08-04 | 四川农业大学 | Potato isolated culture one-step seedling culture medium and optimization method and seedling method thereof |
| CN102138531A (en) * | 2011-04-26 | 2011-08-03 | 马宗新 | Rapid propagation technology and culture medium composition of Fuyang sweet potato 24 virus-free plantlet |
| CN102577960A (en) * | 2012-02-27 | 2012-07-18 | 江阴沃土农业生物科技开发有限公司 | Tissue culture method of potatoes |
| CN106035083A (en) * | 2016-06-01 | 2016-10-26 | 遵义县华富农业生物科技有限公司 | Tissue culture method for rapid detoxification of purple potatoes |
-
2018
- 2018-09-30 CN CN201811155517.2A patent/CN109042337A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH04287623A (en) * | 1991-03-15 | 1992-10-13 | Mitsui Petrochem Ind Ltd | Method for proliferating bulbs by plant tissue culture |
| CN101790935A (en) * | 2010-03-31 | 2010-08-04 | 四川农业大学 | Potato isolated culture one-step seedling culture medium and optimization method and seedling method thereof |
| CN102138531A (en) * | 2011-04-26 | 2011-08-03 | 马宗新 | Rapid propagation technology and culture medium composition of Fuyang sweet potato 24 virus-free plantlet |
| CN102577960A (en) * | 2012-02-27 | 2012-07-18 | 江阴沃土农业生物科技开发有限公司 | Tissue culture method of potatoes |
| CN106035083A (en) * | 2016-06-01 | 2016-10-26 | 遵义县华富农业生物科技有限公司 | Tissue culture method for rapid detoxification of purple potatoes |
Non-Patent Citations (4)
| Title |
|---|
| AHMET METIN KUMLAY等: ""Callus induction, shoot proliferation and root regeneration of potato ( Solanum tuberosum L.) stem node and leaf explants under long-day conditions"", 《BIOTECHNOLOGY & BIOTECHNOLOGICAL EQUIPMENT》 * |
| 张翔龙: "《中学生物知识表解》", 30 September 1986, 重庆出版社 * |
| 方贯娜等: ""马铃薯愈伤组织再生体系的研究进展"", 《中国马铃薯》 * |
| 李义: "《植物快繁技术》", 31 October 2016, 四川大学出版社 * |
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