CN108450332A - A kind of Chinese toon stem section method for tissue culture - Google Patents
A kind of Chinese toon stem section method for tissue culture Download PDFInfo
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- CN108450332A CN108450332A CN201810336967.5A CN201810336967A CN108450332A CN 108450332 A CN108450332 A CN 108450332A CN 201810336967 A CN201810336967 A CN 201810336967A CN 108450332 A CN108450332 A CN 108450332A
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- China
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- stem section
- culture
- chinese toon
- seedling
- 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
-
- 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 present invention provides a kind of Chinese toon stem section method for tissue culture, belongs to field of plant tissue culture.A kind of Chinese toon stem section method for tissue culture provided washes seedling by explant selection, explant processing, stem section proliferation, lateral bud sprouting, strong plantlets and rootage, hardening, bottle outlet, sterilisation step carries out tissue cultivating and seedling.The present invention realizes the tissue cultures high-volume nursery of Chinese toon, and the Chinese toon shape and the speed of growth of cultivation are presented unanimously, improves Chinese toon stem section tissue culture of sprout growth situation, improves plantation survival rate.
Description
Technical field
The invention belongs to field of plant tissue culture, and in particular to a kind of Chinese toon stem section method for tissue culture.
Background technology
Chinese toon also known as Chinese toon bud, fragrant pile crown, bright red Ailanthus altissima, Chinese toon day etc. also troat seedling in Anhui province；Root has two layers
Skin, also known as the bark of the root or stem of the tree of heaven, it is native to China, it is distributed in the extensive area in the Changjiang river north and south, is Meliaceae.Deciduous tree, dioecism, leaf
In even number pinnate compound leaf, panicle, hermaphrodite flower white, fruit is oval capsule, aliform seed, and seed can be bred.Tree
Body is tall and big, except for Chinese toon bud in addition to edible and the preferred seeds of afforestation.Ancient times Chinese toon is referred to as Chinese toon, and tree-of-heaven is referred to as the tree of heaven.Chinese
For a long time Cheng Xi, Han dynasty just spread all on both sides of the Changjiang River edible Chinese toon.Chinese toon bud is full of nutrition, and have dietary function, cure mainly catch cold,
Arthralgia pain due to rheumatism, stomachache, dysentery etc..In actual production, the conventional method of production Chinese toon seedling is, 1, bred with root system, but by
Yu Genliang is very little, and nursery amount is relatively low, and can only breed primary, developmental retardation every year；2, with seminal propagation seedling, can make single plant it
Between external morphology and outer shape and the speed of growth inconsistency is presented, cause quality, nutrient content difference too big.
Invention content
Present invention aim to address above-mentioned deficiencies, provide a kind of Chinese toon stem section method for tissue culture.
To achieve the above object, technical solution of the invention is：A kind of Chinese toon stem section method for tissue culture, feature
It is to carry out as follows：
The first step, explant are chosen：The breeding maternal plant of growth selection stalwartness is shadowed branch or side in calm bright day gas clip
Branch carries out refrigeration moisturizing and takes back laboratory treatment at once.
Second step, explant processing：Adopt the vegetable material come and remove no part, it would be desirable to set water tap
Under, it sets in beaker and is rinsed about 1 hour with flowing water, dry the moisture content on surface, wrapped up with clean-keeping paper；Treated, and explant is first
It with sterile water wash 2 ~ 3 times, is impregnated 30 seconds with 75% alcohol, afterwards with sterile water wash 2~3 times, then with the mercuric chloride leaching of 0.1% concentration
Steep 20min, then with sterile water wash 4~5 times, it is spare.
Third walks, stem section proliferation：Aseptically, the Chinese toon stem section disinfected is clamped with tweezers, first with dissection
Knife cuts off blade, then is cut off from germination point lower section about 1.5cm or so, top 0.5cm or so, and each stem section 1.5 is retained
~2.0cm long, access fill in the culture bottle for increasing stem culture medium, and each culture bottle accesses 2 ~ 3 germination points；By the culture of inoculation
Bottle is placed in temperature, light irradiation time be 14 hours/day, intensity of illumination be 2000lux~
It is cultivated under the conditions of 2500lux, promotes stem section growth.
4th step, lateral bud are sprouted：Stem section elongated after growth is transferred, same length stem section is cut and is put into and fill side
In the culture bottle of bud germination medium, each culture bottle connects 2 ~ 3 germination points, and it is 23~27 DEG C that the culture bottle of inoculation, which is placed in temperature,
Clean environment in, cultivated under the conditions of light irradiation time is 14 hours/day, intensity of illumination is 2000lux~2500lux, make it
Sprout lateral bud from base contact.
5th step, strong plantlets and rootage：It is 3~4cm length to select sprout health, free of contamination lateral bud or terminal bud, length, is put
Enter in root media, is 2000lux~2500lux in intensity of illumination, light irradiation time is 14 hours/day, culture 30 days~45
It.
6th step, hardening：Seedling is taken exercise under 15 DEG C~35 DEG C of temperature, illumination 2000lux~20000lux range of condition
After 15 days~20 days.
7th step, bottle outlet wash seedling：Bottleneck is opened, the seedling after hardening is gently pressed from both sides out with tweezers, uses flowing water
Under cleaned, clean root attachment culture medium.
8th step, disinfection：It will be grown in cleaned seedling transfer to sterile-processed soil.
The increasing stem medium component group by weight is divided into：The increasing stem medium component group by weight is divided into：MS
Culture medium is 1000 parts, gibberellin is 0.5 part of mixing, and it is 5.8 to add NaOH and adjust pH value；The lateral bud germination medium
Ingredient group by weight is divided into：MS culture mediums are 1000 parts, methyl α-naphthyl acetate is 1 part, the basic element of cell division is 0.5 part of mixing, are added
It is 5.8 that NaOH, which adjusts pH value,；The culture of rootage based component group by weight is divided into：MS culture mediums are 500 parts, indolebutyric acid is
1 part, sucrose be 30 parts, add NaOH adjust pH value be 5.8.
A kind of Chinese toon stem section method for tissue culture of the offer of the present invention is passed through by long-term experiment and real work tissue culture
It tests, by the mating reaction of culture medium each component and each step, realizes the tissue cultures high-volume nursery of Chinese toon, improve and educate
Seedling efficiency, the Chinese toon shape and the speed of growth of cultivation present consistent, and the Chinese toon quality that grows up to, nutrient content are high and consistent；Chinese toon
The bottle seedling stem section speed of growth is fast, and the lateral bud quantity of sprouting is more, and the seedling time is short, and young shoot cannot directly be formed by overcoming callus
Defect improves Chinese toon stem section tissue cultures growth of seedling situation, improves plantation survival rate, rooting rate have currently reached 60% with
On.
Specific implementation mode
A kind of Chinese toon stem section method for tissue culture of the offer of the present invention is specifically carried out by following examples.
The first step, explant are chosen：The breeding maternal plant of growth selection stalwartness, calm bright day gas clip shadow branch or
Side shoot carries out refrigeration moisturizing and takes back laboratory treatment at once.
Second step, explant processing：Adopt the vegetable material come and remove no part, it would be desirable to set water tap
Under, it sets in beaker and is rinsed about 1 hour with flowing water, dry the moisture content on surface, wrapped up with clean-keeping paper；Treated, and explant is first
It with sterile water wash 2 ~ 3 times, is impregnated 30 seconds with 75% alcohol, afterwards with sterile water wash 2~3 times, then with the mercuric chloride leaching of 0.1% concentration
Steep 20min, then with sterile water wash 4~5 times, it is spare.
Third walks, stem section proliferation：Aseptically, the Chinese toon stem section disinfected is clamped with tweezers, first with dissection
Knife cuts off blade, then is cut off from germination point lower section about 1.5cm or so, top 0.5cm or so, and each stem section 1.5 is retained
~2.0cm long, access fill in the culture bottle for increasing stem culture medium, and each culture bottle accesses 2 ~ 3 germination points；By the culture of inoculation
Bottle is placed in temperature, light irradiation time be 14 hours/day, intensity of illumination be 2000lux~
It is cultivated under the conditions of 2500lux, promotes stem section growth.
4th step, lateral bud are sprouted：Stem section elongated after growth is transferred, same length stem section is cut and is put into and fill side
In the culture bottle of bud germination medium, each culture bottle connects 2 ~ 3 germination points, and it is 23~27 DEG C that the culture bottle of inoculation, which is placed in temperature,
Clean environment in, cultivated under the conditions of light irradiation time is 14 hours/day, intensity of illumination is 2000lux~2500lux, make it
Sprout lateral bud from base contact.
5th step, strong plantlets and rootage：It is 3~4cm length to select sprout health, free of contamination lateral bud or terminal bud, length, is put
Enter in root media, is 2000lux~2500lux in intensity of illumination, light irradiation time is 14 hours/day, culture 30 days~45
It.
6th step, hardening：Seedling is taken exercise under 15 DEG C~35 DEG C of temperature, illumination 2000lux~20000lux range of condition
After 15 days~20 days.
7th step, bottle outlet wash seedling：Bottleneck is opened, the seedling after hardening is gently pressed from both sides out with tweezers, uses flowing water
Under cleaned, clean root attachment culture medium.
8th step, disinfection：It will be grown in cleaned seedling transfer to sterile-processed soil.
The increasing stem medium component group by weight is divided into：The increasing stem medium component group by weight is divided into：MS
Culture medium is 1000 parts, gibberellin is 0.5 part of mixing, and it is 5.8 to add NaOH and adjust pH value；The lateral bud germination medium
Ingredient group by weight is divided into：MS culture mediums are 1000 parts, methyl α-naphthyl acetate is 1 part, the basic element of cell division is 0.5 part of mixing, are added
It is 5.8 that NaOH, which adjusts pH value,；The culture of rootage based component group by weight is divided into：MS culture mediums are 500 parts, indolebutyric acid is
1 part, sucrose be 30 parts, add NaOH adjust pH value be 5.8.
The cultivation results of Shucheng County, Anhui Province Agricultural Technological Garden tissue culture planting base：Chinese toon tissue culture method provided by the invention
The Chinese toon bottle seedling stem section speed of growth cultivated is fast, will start within 2~3 days to grow, the lateral bud quantity of sprouting is more, and quantity is 5~6
A, the seedling time is short, and rooting rate has currently reached 60% or more；Young shoot defect cannot directly be formed by overcoming callus, be improved
Plant survival rate.
Above-described embodiment is the technical approach and technical characterstic for illustrating the present invention, in the art common the purpose is to allow
Technical staff cans understand the content of the present invention and implement it accordingly, and it is not intended to limit the scope of the present invention.Every
The equivalent changes or modifications made according to the essence of the content of present invention should all cover within the scope of the present invention.
Claims (2)
1. a kind of Chinese toon stem section method for tissue culture, it is characterised in that：It carries out as follows：
The first step, explant are chosen：The breeding maternal plant of growth selection stalwartness is shadowed branch or side in calm bright day gas clip
Branch carries out refrigeration moisturizing and takes back laboratory treatment at once；
Second step, explant processing：Adopt the vegetable material come and remove no part, it would be desirable to set under water tap, set
It is rinsed about 1 hour with flowing water in beaker, dries the moisture content on surface, wrapped up with clean-keeping paper；Treated, and explant first uses nothing
Bacterium water cleans 2 ~ 3 times, is impregnated 30 seconds with 75% alcohol, uses sterile water wash afterwards 2~3 times, then is impregnated with the mercuric chloride of 0.1% concentration
20min, then with sterile water wash 4~5 times, it is spare；
Third walks, stem section proliferation：Aseptically, the Chinese toon stem section disinfected is clamped with tweezers, it first will with scalpel
Blade is cut off, then from germination point lower section about 1.5cm or so, top 0.5cm or so cut off, retain each stem section 1.5~
2.0cm long, access fill in the culture bottle for increasing stem culture medium, and each culture bottle accesses 2 ~ 3 germination points；By the culture bottle of inoculation
Being placed in temperature is in 23~27 DEG C of clean environment, light irradiation time be 14 hours/day, intensity of illumination be 2000lux~
It is cultivated under the conditions of 2500lux, promotes stem section growth；
4th step, lateral bud are sprouted：Stem section elongated after growth is transferred, same length stem section is cut and is put into and fill lateral bud and sprout
In the culture bottle for sending out culture medium, each culture bottle connects 2 ~ 3 germination points, and it is 23~27 DEG C clean that the culture bottle of inoculation, which is placed in temperature,
In net environment, is cultivated under the conditions of light irradiation time is 14 hours/day, intensity of illumination is 2000lux~2500lux, make it from base
Portion's contact sprouts lateral bud；
5th step, strong plantlets and rootage：It is 3~4cm length to select sprout health, free of contamination lateral bud or terminal bud, length, is put into life
It is 2000lux~2500lux in intensity of illumination, light irradiation time is 14 hours/day, is cultivated 30 days~45 days in root culture medium；
6th step, hardening：Seedling is taken exercise 15 days under 15 DEG C~35 DEG C of temperature, illumination 2000lux~20000lux range of condition
After~20 days；
7th step, bottle outlet wash seedling：Open bottleneck, the seedling after hardening is gently pressed from both sides out with tweezers, under flowing water into
Row cleaning, cleans the culture medium of root attachment；
8th step, disinfection：It will be grown in cleaned seedling transfer to sterile-processed soil.
2. a kind of Chinese toon stem section method for tissue culture as described in claim 1, it is characterised in that：The increasing stem culture medium at
Group by weight is divided to be divided into：MS culture mediums are 1000 parts, gibberellin is 0.5 part of mixing, and it is 5.8 to add NaOH and adjust pH value；Institute
The lateral bud germination medium ingredient group by weight stated is divided into：MS culture mediums are 1000 parts, methyl α-naphthyl acetate is 1 part, the basic element of cell division is
0.5 part of mixing, it is 5.8 to add NaOH and adjust pH value；The culture of rootage based component group by weight is divided into：MS culture mediums are
500 parts, indolebutyric acid be 1 part, sucrose is 30 parts, add NaOH adjust pH value be 5.8.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810336967.5A CN108450332A (en) | 2018-04-16 | 2018-04-16 | A kind of Chinese toon stem section method for tissue culture |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810336967.5A CN108450332A (en) | 2018-04-16 | 2018-04-16 | A kind of Chinese toon stem section method for tissue culture |
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| CN108450332A true CN108450332A (en) | 2018-08-28 |
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| CN201810336967.5A Pending CN108450332A (en) | 2018-04-16 | 2018-04-16 | A kind of Chinese toon stem section method for tissue culture |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110583487A (en) * | 2019-10-24 | 2019-12-20 | 安徽东方金桥农林科技股份有限公司 | Tissue culture method of Taihe Chinese toon |
| CN111194694A (en) * | 2020-03-04 | 2020-05-26 | 聊城大学 | Tissue culture rapid propagation method of new plant variety chat toona sinensis |
Citations (5)
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| CN101491215A (en) * | 2009-02-19 | 2009-07-29 | 周玉玲 | Chinese toon tissue-culture quick propagation technique and culture medium proportion |
| CN102138532A (en) * | 2011-04-26 | 2011-08-03 | 马宗新 | Technique for breeding seedlings of Taihe tribute Chinese toon by tissue culture |
| CN102577944A (en) * | 2011-12-21 | 2012-07-18 | 广西金宏昕生物科技有限公司 | Method for high-efficient culture of clonal tissue culture seedlings of Chinese toons |
| CN103621400A (en) * | 2012-08-20 | 2014-03-12 | 王宝雪 | Toona sinensis bud soilless cultivation method |
| CN104756866A (en) * | 2015-03-18 | 2015-07-08 | 胡莹莹 | Cuttage rapid propagation method of test-tube plantlet of toona sinensis |
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2018
- 2018-04-16 CN CN201810336967.5A patent/CN108450332A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101491215A (en) * | 2009-02-19 | 2009-07-29 | 周玉玲 | Chinese toon tissue-culture quick propagation technique and culture medium proportion |
| CN102138532A (en) * | 2011-04-26 | 2011-08-03 | 马宗新 | Technique for breeding seedlings of Taihe tribute Chinese toon by tissue culture |
| CN102577944A (en) * | 2011-12-21 | 2012-07-18 | 广西金宏昕生物科技有限公司 | Method for high-efficient culture of clonal tissue culture seedlings of Chinese toons |
| CN103621400A (en) * | 2012-08-20 | 2014-03-12 | 王宝雪 | Toona sinensis bud soilless cultivation method |
| CN104756866A (en) * | 2015-03-18 | 2015-07-08 | 胡莹莹 | Cuttage rapid propagation method of test-tube plantlet of toona sinensis |
Non-Patent Citations (1)
| Title |
|---|
| 许丽琼 涂炳坤: "香椿茎段组织培养和再生技术研究", 《华中农业大学学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110583487A (en) * | 2019-10-24 | 2019-12-20 | 安徽东方金桥农林科技股份有限公司 | Tissue culture method of Taihe Chinese toon |
| CN111194694A (en) * | 2020-03-04 | 2020-05-26 | 聊城大学 | Tissue culture rapid propagation method of new plant variety chat toona sinensis |
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Application publication date: 20180828 |