CN108064696A - A kind of blueberry tissue cultural method - Google Patents
A kind of blueberry tissue cultural method Download PDFInfo
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- CN108064696A CN108064696A CN201810049740.2A CN201810049740A CN108064696A CN 108064696 A CN108064696 A CN 108064696A CN 201810049740 A CN201810049740 A CN 201810049740A CN 108064696 A CN108064696 A CN 108064696A
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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 discloses a kind of blueberry tissue cultural methods, it is first inoculated in the first culture medium as explant using shoot stem with bud and carries out adventitious bud induction culture, it is inoculated into after the completion of adventitious bud induction culture in the second culture medium and carries out Multiplying culture, it is inoculated into after the completion of Multiplying culture in the 3rd culture medium and carries out culture of rootage, the tissue-cultured seedling that will finally take root is placed at room temperature in daylight hardening 79 days, transplanting is into vermiculite and perlite mixed-matrix after being sterilized with 1000 times of carbendazim solutions, epiphragma or spraying and moisturizing after transplanting.Breeding culture is carried out to blueberry using the method for the present invention, success rate is up to 83.4%, and tissue-cultured seedling is grown, and survival rate is up to 93.8%, and well developed root system, robust plant, growing way are strong.
Description
Technical field
The present invention relates to plant tissue Cultivating techniques fields, and in particular to a kind of blueberry tissue cultural method.
Background technology
Blueberry also known as cowberry, blue berry, Ericaceae cowberry platymiscium, are perennial fallen leaves or evergreen shrubs fruit tree.Its
Berry contains abundant anthocyanidin, and having prevents cerebral nervous system aging, anticancer, softening blood vessel, enhancing human organism's immunity etc.
Function regards as one of five big healthy food by international food and agricultural organization, possesses the good reputation of " gold berry ".Blueberry is as a kind of new
Type fruit, economic value and healthy nutritive value are higher, have wide exploitation prospect.In recent years, blueberry cultivated area
It is growing, but conventional cuttage and seedling culture mode cultivation cycle is long, of high cost, reproductive efficiency is low, and tissue culture technique can be shown
The cultivation cycle for shortening blueberry seedling is write, breeding coefficient is high, can cultivate virus-free plant.
The content of the invention
In view of the above shortcomings of the prior art, the object of the present invention is to provide a kind of blueberry tissue cultural methods.
The present invention adopts the technical scheme that：
A kind of blueberry tissue cultural method, comprises the following steps：
(1) explant is chosen, chooses and gives birth to shoot stem with bud then, rinsed well with tap water and use aseptic water washing again
2-3 times, 10-20s then is impregnated with 75% alcohol, is rinsed well with tap water and uses aseptic water washing again 2-3 times, Ran Houyong
0.1% mercury chloride or 2.0% hypochlorous acid, which are received, impregnates 7-10min, finally with aseptic water washing 2-3 times；
(2) explant is inoculated into the first culture medium and carries out adventitious bud induction culture, first culture medium is：1/
2MS、ZT 1mg/L、IBA0.5mg/L、NAA0.05mg/L；
(3) it is inoculated into after the completion of adventitious bud induction culture in the second culture medium and carries out Multiplying culture, second culture medium
For：1/2MS、BA0.5mg/L、6-BA1mg/L、NAA0.05mg/L；
(4) it is inoculated into after the completion of Multiplying culture in the 3rd culture medium and carries out culture of rootage, the 3rd culture medium is：1/
4MS、IBA 0.5mg/L、ABT0.1mg/L、NAA0.1mg/L；
(5) tissue-cultured seedling that will take root is placed at room temperature in daylight hardening 7-9 days, is moved after being sterilized with 1000 times of carbendazim solutions
It plants into vermiculite and perlite mixed-matrix, epiphragma or spraying and moisturizing after transplanting, the mass ratio of the vermiculite and perlite is 1:
1。
Further, the condition of culture is：Day temperature is 24 ± 1 DEG C, 20 ± 1 DEG C of night temperatures, intensity of illumination
For 1500-2000lx, daily light application time is 12-14h.
Further, the adventitious bud induction culture time is 25-30d.
Further, the Multiplying culture time is 25-30d.
Further, the culture of rootage time is 10-15d.
It is an advantage of the invention that：Breeding culture is carried out to blueberry using the method for the present invention, success rate is up to 83.4%, group
Training seedling is grown, and survival rate is up to 93.8%, and well developed root system, robust plant, growing way are strong.
Specific embodiment
For a further understanding of the present invention, the preferred embodiment of the invention is described with reference to embodiment, still
It should be appreciated that these descriptions are simply for the feature and advantage that further illustrate the present invention rather than to the claims in the present invention
Limitation.
Embodiment 1
A kind of blueberry tissue cultural method, comprises the following steps：
(1) explant is chosen, chooses and gives birth to shoot stem with bud then, rinsed well with tap water and use aseptic water washing again
2 times, 10s then is impregnated with 75% alcohol, is rinsed well with tap water and uses aseptic water washing again 2 times, then with 0.1% chlorination
Mercury impregnates 7min, finally with aseptic water washing 2 times；
(2) bring explant into laboratory to cultivate, condition of culture is：Day temperature is 23 DEG C, 19 DEG C of night temperatures,
Intensity of illumination is 1500lx, and daily light application time is 12h, and explant is inoculated into progress adventitious bud in the first culture medium first lures
Culture 25d is led, first culture medium is：1/2MS、ZT 1mg/L、IBA0.5mg/L、NAA0.05mg/L；
(3) it is inoculated into after the completion of adventitious bud induction culture in the second culture medium and carries out Multiplying culture 25d, second culture
Base is：1/2MS、BA0.5mg/L、6-BA1mg/L、NAA0.05mg/L；
(4) progress culture of rootage 10d, the 3rd culture medium in the 3rd culture medium are inoculated into after the completion of Multiplying culture is：
1/4MS、IBA 0.5mg/L、ABT0.1mg/L、NAA0.1mg/L；
(5) tissue-cultured seedling that will take root is placed at room temperature in daylight hardening 7 days, is transplanted after being sterilized with 1000 times of carbendazim solutions
Into vermiculite and perlite mixed-matrix, epiphragma or spraying and moisturizing after transplanting, the mass ratio of the vermiculite and perlite is 1:1.
The present embodiment explant success rate is 80.3%, and tissue culture shoot survival percent is 91.5%.
Embodiment 2
A kind of blueberry tissue cultural method, comprises the following steps：
(1) explant is chosen, chooses and gives birth to shoot stem with bud then, rinsed well with tap water and use aseptic water washing again
3 times, 15s then is impregnated with 75% alcohol, is rinsed well with tap water and uses aseptic water washing again 3 times, then with 0.1% chlorination
Mercury impregnates 8min, finally with aseptic water washing 3 times；
(2) bring explant into laboratory to cultivate, condition of culture is：Day temperature is 24 DEG C, 20 DEG C of night temperatures,
Intensity of illumination is 1800lx, and daily light application time is 13h, and explant is inoculated into progress adventitious bud in the first culture medium first lures
Culture 28d is led, first culture medium is：1/2MS、ZT 1mg/L、IBA0.5mg/L、NAA0.05mg/L；
(3) it is inoculated into after the completion of adventitious bud induction culture in the second culture medium and carries out Multiplying culture 28d, second culture
Base is：1/2MS、BA0.5mg/L、6-BA1mg/L、NAA0.05mg/L；
(4) progress culture of rootage 13d, the 3rd culture medium in the 3rd culture medium are inoculated into after the completion of Multiplying culture is：
1/4MS、IBA 0.5mg/L、ABT0.1mg/L、NAA0.1mg/L；
(5) tissue-cultured seedling that will take root is placed at room temperature in daylight hardening 8 days, is transplanted after being sterilized with 1000 times of carbendazim solutions
Into vermiculite and perlite mixed-matrix, epiphragma or spraying and moisturizing after transplanting, the mass ratio of the vermiculite and perlite is 1:1.
The present embodiment explant success rate is 82.8%, and tissue culture shoot survival percent is 93.5%.
Embodiment 3
A kind of blueberry tissue cultural method, comprises the following steps：
(1) explant is chosen, chooses and gives birth to shoot stem with bud then, rinsed well with tap water and use aseptic water washing again
3 times, 20s then is impregnated with 75% alcohol, is rinsed well with tap water and uses aseptic water washing again 3 times, then with 2.0% chlorine
Acid, which is received, impregnates 10min, finally with aseptic water washing 3 times；
(2) bring explant into laboratory to cultivate, condition of culture is：Day temperature is 25 DEG C, 21 DEG C of night temperatures,
Intensity of illumination is 2000lx, and daily light application time is 14h, and explant is inoculated into progress adventitious bud in the first culture medium first lures
Culture 30d is led, first culture medium is：1/2MS、ZT 1mg/L、IBA0.5mg/L、NAA0.05mg/L；
(3) it is inoculated into after the completion of adventitious bud induction culture in the second culture medium and carries out Multiplying culture 30d, second culture
Base is：1/2MS、BA0.5mg/L、6-BA1mg/L、NAA0.05mg/L；
(4) progress culture of rootage 15d, the 3rd culture medium in the 3rd culture medium are inoculated into after the completion of Multiplying culture is：
1/4MS、IBA 0.5mg/L、ABT0.1mg/L、NAA0.1mg/L；
(5) tissue-cultured seedling that will take root is placed at room temperature in daylight hardening 9 days, is transplanted after being sterilized with 1000 times of carbendazim solutions
Into vermiculite and perlite mixed-matrix, epiphragma or spraying and moisturizing after transplanting, the mass ratio of the vermiculite and perlite is 1:1.
The present embodiment explant success rate is 82.1%, and tissue culture shoot survival percent is 92.8%.
Claims (5)
1. a kind of blueberry tissue cultural method, it is characterised in that comprise the following steps：
(1) explant is chosen, chooses and gives birth to shoot stem with bud then, is rinsed well with tap water and uses aseptic water washing 2-3 again
Time, 10-20s then is impregnated with 75% alcohol, is rinsed well with tap water and uses aseptic water washing again 2-3 times, then with 0.1%
Mercury chloride or 2.0% hypochlorous acid, which are received, impregnates 7-10min, finally with aseptic water washing 2-3 times；
(2) explant is inoculated into the first culture medium and carries out adventitious bud induction culture, first culture medium is：1/2MS、ZT
1mg/L、IBA0.5mg/L、NAA0.05mg/L；
(3) it is inoculated into after the completion of adventitious bud induction culture in the second culture medium and carries out Multiplying culture, second culture medium is：1/
2MS、BA0.5mg/L、6-BA1mg/L、NAA0.05mg/L；
(4) it is inoculated into after the completion of Multiplying culture in the 3rd culture medium and carries out culture of rootage, the 3rd culture medium is：1/4MS、
IBA 0.5mg/L、ABT0.1mg/L、NAA0.1mg/L；
(5) tissue-cultured seedling that will take root is placed at room temperature in daylight hardening 7-9 days, transplanted after being sterilized with 1000 times of carbendazim solutions to
In vermiculite and perlite mixed-matrix, epiphragma or spraying and moisturizing after transplanting, the mass ratio of the vermiculite and perlite is 1:1.
2. blueberry tissue cultural method according to claim 1, it is characterised in that the condition of culture is：Day temperature
For 24 ± 1 DEG C, 20 ± 1 DEG C of night temperatures, intensity of illumination 1500-2000lx, daily light application time is 12-14h.
3. blueberry tissue cultural method according to claim 1, it is characterised in that the adventitious bud induction culture time is 25-
30d。
4. blueberry tissue cultural method according to claim 1, it is characterised in that the Multiplying culture time is 25-30d.
5. blueberry tissue cultural method according to claim 1, it is characterised in that the culture of rootage time is 10-15d.
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| CN201810049740.2A CN108064696A (en) | 2018-01-18 | 2018-01-18 | A kind of blueberry tissue cultural method |
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| CN201810049740.2A CN108064696A (en) | 2018-01-18 | 2018-01-18 | A kind of blueberry tissue cultural method |
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| CN108064696A true CN108064696A (en) | 2018-05-25 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114287341A (en) * | 2021-11-15 | 2022-04-08 | 安徽师范大学 | Method for culturing blueberry tissue |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20120102300A (en) * | 2011-03-08 | 2012-09-18 | 충청북도 (관리부서:충청북도 농업기술원) | Method for plant formation of blueberry cv. bluegold, elizabeth, darrow, woodard or tifblue through apical meristem culture |
| CN106342688A (en) * | 2016-08-29 | 2017-01-25 | 盛林蓝莓集团股份有限公司 | Virus-free seedling method of blueberry germchit |
-
2018
- 2018-01-18 CN CN201810049740.2A patent/CN108064696A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20120102300A (en) * | 2011-03-08 | 2012-09-18 | 충청북도 (관리부서:충청북도 농업기술원) | Method for plant formation of blueberry cv. bluegold, elizabeth, darrow, woodard or tifblue through apical meristem culture |
| CN106342688A (en) * | 2016-08-29 | 2017-01-25 | 盛林蓝莓集团股份有限公司 | Virus-free seedling method of blueberry germchit |
Non-Patent Citations (5)
| Title |
|---|
| 姜冬等: "蓝莓的组织培养及快速繁殖", 《中国林副特产》 * |
| 宁志怨等: "蓝葛丛生芽的诱导及植株再生", 《分子植物育种》 * |
| 杨春梅等: "江南越桔组织培养和快繁技术研究", 《西南农业学报》 * |
| 樊庆杰等: "北高灌蓝莓的离体快速繁殖研究", 《中国新技术新产品》 * |
| 陈平芬等: "南高丛蓝莓良种薄雾组培快繁技术研究", 《现代农业科技》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114287341A (en) * | 2021-11-15 | 2022-04-08 | 安徽师范大学 | Method for culturing blueberry tissue |
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