CN107667866A - The cultural method of common calla tissue-cultured seedling - Google Patents
The cultural method of common calla tissue-cultured seedling Download PDFInfo
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- CN107667866A CN107667866A CN201711119709.3A CN201711119709A CN107667866A CN 107667866 A CN107667866 A CN 107667866A CN 201711119709 A CN201711119709 A CN 201711119709A CN 107667866 A CN107667866 A CN 107667866A
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- cultured seedling
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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 cultural method of common calla tissue-cultured seedling, comprise the following steps：Step 1: it is explant to choose common calla main bud or lateral bud, after use quality fraction is 65% alcohol disinfecting, with aseptic water washing 12 minutes, then it is placed in the liquor natrii hypochloritis that mass fraction is 15 25% and sterilizes 5 minutes, after aseptic water washing 46 minutes, explant is obtained；Step 2: explant is placed in into culture in inducing culture extremely forms adventitious bud；Step 3: it is numerous that adventitious bud is moved into progress subculture expansion on subculture medium；Step 4: after the budlet on subculture medium is grown to more than 2cm, move it to and culture of rootage and strong sprout are carried out in Rooting and hardening-off culture base；Step 5: when the tissue-cultured seedling length on Rooting and hardening-off culture base to 23 leaves, hardening 47 days, when growing 35 leaves to training tissue culture seedling, you can transplanted.It is an object of the invention to provide a kind of cultural method of common calla tissue-cultured seedling.
Description
Technical field
The present invention relates to field of plant tissue culture.A kind of it is more particularly related to training of common calla tissue-cultured seedling
The method of supporting.
Background technology
Common calla originates in South Africa and is also known as Alocasia ordora, water arum, Alocasia amazonica, Araeceae horseshoe Nelumbo, is flowering bulb, meat
Stem tuber is loose, is taken root at section, well developed root system is sturdy, and its flower pattern is spadix, is wrapped in the Buddhist flame of the different color such as yellow, red, powder
In bud, different cultivars system is formed.Calla lily both considerable leaf or considerable flower, have higher ornamental value.Common calla it is numerous
Usual two methods are grown, traditional method is carried out using the mitogenetic method of stem tuber, and this method breeding is slow, survival rate is low, the production cycle
Long, easily variation, breeding coefficient are small；Another method is group culturation rapid propagating technology, but because different kinds is in induction, subculture expansion
The numerous, stage of taking root requires different culture mediums, therefore each designs and varieties are required for the special training of itself suitable fast numerous needs
Support base.
In a broad sense, Plant Tissue Breeding concept is called cultured in vitro, refers to the tissue isolated and suited the requirements from plant.
Organ or cell, protoplast etc., by sterile working, cultivated under manual control condition to obtain the complete plant of regeneration
The technology of other products of strain or production with economic value.And in the narrow sense, Plant Tissue Breeding concept refers to each with plant
Portion of tissue, such as forming layer, parenchymal tissue, mesophyll tissue, endosperm carry out culture and obtain regeneration plant, also refer in incubation
In the culture of callus is produced from each organ, callus again by being differentiated to form aftergrowth again.
The content of the invention
It is an object of the invention to provide a kind of cultural method of common calla tissue-cultured seedling.
In order to realize according to object of the present invention and further advantage, there is provided a kind of culture side of common calla tissue-cultured seedling
Method, comprise the following steps：
Step 1: it is explant to choose common calla main bud or lateral bud, use quality fraction is use after 65% alcohol disinfecting
Aseptic water washing 1-2 minutes, then be placed in the liquor natrii hypochloritis that mass fraction is 15-25% and sterilize 5 minutes, rushed with sterilized water
After washing 4-6 minutes, explant is obtained；
Step 2: the explant is placed in into culture in inducing culture extremely forms adventitious bud, wherein, the Fiber differentiation
Base includes：MS, 2-3mg/L 6-benzyladenine, 0.1-0.2mg/L methyl α-naphthyl acetate, 0.2-0.6mg/L gibberellin, 20g/L
The agar of sucrose and 12g/L；
Step 3: by the adventitious bud move on subculture medium carry out subculture expand it is numerous, wherein, the subculture medium bag
Include：MS, 2-3mg/L 6-benzyladenine, 0.2-0.6mg/L gibberellin, 0.5-2.0mg/L Methotrexate, 22g/L sugarcanes
The agar of sugar and 15g/L；
Step 4: after the budlet on the subculture medium is grown to more than 2cm, move it in Rooting and hardening-off culture base
Culture of rootage and strong sprout are carried out, wherein, the Rooting and hardening-off culture base includes：1/2MS, 3-5mg/L 6-benzyladenine,
0.4-0.6mg/L methyl α-naphthyl acetate, 0.2-0.4mg/L gibberellin, 30g/L sucrose and 18g/L agar；
Step 5: when the tissue-cultured seedling length on the Rooting and hardening-off culture base to 2-3 piece leaves, hardening 4-7 days, to tissue-cultured seedling
When hardening grows 3-5 piece leaves, you can transplanted.
Preferably, in the cultural method of described common calla tissue-cultured seedling, when culture is to adventitious bud in the step 2, its
Cultivation temperature is 23-28 DEG C, intensity of illumination 1200-1800lux, and light application time is 8-10 hours/day.
Preferably, in the cultural method of described common calla tissue-cultured seedling, when progress subculture expands numerous in the step 3, its
Cultivation temperature is 23-28 DEG C, intensity of illumination 1200-2000lux, and light application time is 8-12 hours/day.
Preferably, in the cultural method of described common calla tissue-cultured seedling, the step 4 carries out culture of rootage and strong sprout
When, its cultivation temperature is 23-27 DEG C, intensity of illumination 1200-2000lux, and light application time is 8-12 hours/day.
Preferably, in the cultural method of described common calla tissue-cultured seedling, the inducing culture, subculture medium and life
The initial pH value of root strong seedling culture base is 5-6.
The present invention common calla tissue-cultured seedling cultural method improve common calla outside shade cultivation survival rate, will take root and
Strong sprout merges culture, shortens cultivation cycle, has saved cost, improved economic benefit.
Further advantage, target and the feature of the present invention embodies part by following explanation, and part will also be by this
The research and practice of invention and be understood by the person skilled in the art.
Embodiment
<Embodiment 1>
A kind of cultural method of common calla tissue-cultured seedling, comprises the following steps：
Step 1: it is explant to choose common calla main bud or lateral bud, use quality fraction is use after 65% alcohol disinfecting
Aseptic water washing 1 minute, then be placed in the liquor natrii hypochloritis that mass fraction is 15% and sterilize 5 minutes, divided with aseptic water washing 4
Zhong Hou, obtain explant；
Step 2: the explant is placed in into culture in inducing culture extremely forms adventitious bud, wherein, cultivation temperature 23
DEG C, intensity of illumination 1200lux, light application time is 8 hours/day, and the inducing culture includes：MS, 2mg/L 6- benzyl glands
Purine, 0.1mg/L methyl α-naphthyl acetate, 0.2mg/L gibberellin, 20g/L sucrose and 12g/L agar；
Step 3: by the adventitious bud move on subculture medium carry out subculture expand it is numerous, wherein, cultivation temperature be 23 DEG C,
Intensity of illumination is 1200lux, and light application time is 8 hours/day, and the subculture medium includes：MS, 2mg/L 6- benzyl glands are fast
Purine, 0.2mg/L gibberellin, 0.5mg/L Methotrexate, 22g/L sucrose and 15g/L agar；
Step 4: after the budlet on the subculture medium is grown to more than 2cm, move it in Rooting and hardening-off culture base
Culture of rootage and strong sprout are carried out, wherein, cultivation temperature is 23 DEG C, intensity of illumination 1200lux, and light application time is 8 hours/day,
The Rooting and hardening-off culture base includes：1/2MS, 3mg/L 6-benzyladenine, 0.4mg/L methyl α-naphthyl acetate, 0.2mg/L it is red
The agar of mycin, 30g/L sucrose and 18g/L；
Step 5: when the tissue-cultured seedling length on the Rooting and hardening-off culture base to 2 leaves, hardening 4 days, to training tissue culture seedling
When growing 3 leaves, you can transplanted.
Wherein, the initial pH value of the inducing culture, subculture medium and Rooting and hardening-off culture base is 5.
<Embodiment 2>
A kind of cultural method of common calla tissue-cultured seedling, comprises the following steps：
Step 1: it is explant to choose common calla main bud or lateral bud, use quality fraction is use after 65% alcohol disinfecting
Aseptic water washing 1 minute, then be placed in the liquor natrii hypochloritis that mass fraction is 20% and sterilize 5 minutes, divided with aseptic water washing 5
Zhong Hou, obtain explant；
Step 2: the explant is placed in into culture in inducing culture extremely forms adventitious bud, wherein, cultivation temperature 25
DEG C, intensity of illumination 1600lux, light application time is 9 hours/day, and the inducing culture includes：MS, 2.5mg/L 6- benzyls
Adenine, 0.15mg/L methyl α-naphthyl acetate, 0.4mg/L gibberellin, 20g/L sucrose and 12g/L agar；
Step 3: by the adventitious bud move on subculture medium carry out subculture expand it is numerous, wherein, cultivation temperature be 25 DEG C,
Intensity of illumination is 1600lux, and light application time is 10 hours/day, and the subculture medium includes：MS, 2.5mg/L 6- benzyl glands
Purine, 0.4mg/L gibberellin, 1.5mg/L Methotrexate, 22g/L sucrose and 15g/L agar；
Step 4: after the budlet on the subculture medium is grown to more than 2cm, move it in Rooting and hardening-off culture base
Culture of rootage and strong sprout are carried out, wherein, cultivation temperature is 25 DEG C, intensity of illumination 1600lux, and light application time is 10 hours/day,
The Rooting and hardening-off culture base includes：1/2MS, 4mg/L 6-benzyladenine, 0.5mg/L methyl α-naphthyl acetate, 0.3mg/L it is red
The agar of mycin, 30g/L sucrose and 18g/L；
Step 5: when the tissue-cultured seedling length on the Rooting and hardening-off culture base to 2 leaves, hardening 6 days, to training tissue culture seedling
When growing 4 leaves, you can transplanted.
Wherein, the initial pH value of the inducing culture, subculture medium and Rooting and hardening-off culture base is 5.
<Embodiment 3>
A kind of cultural method of common calla tissue-cultured seedling, comprises the following steps：
Step 1: it is explant to choose common calla main bud or lateral bud, use quality fraction is use after 65% alcohol disinfecting
Aseptic water washing 2 minutes, then be placed in the liquor natrii hypochloritis that mass fraction is 25% and sterilize 5 minutes, divided with aseptic water washing 6
Zhong Hou, obtain explant；
Step 2: the explant is placed in into culture in inducing culture extremely forms adventitious bud, wherein, cultivation temperature 28
DEG C, intensity of illumination 1800lux, light application time is 10 hours/day, and the inducing culture includes：MS, 3mg/L 6- benzyls
Adenine, 0.2mg/L methyl α-naphthyl acetate, 0.6mg/L gibberellin, 20g/L sucrose and 12g/L agar；
Step 3: by the adventitious bud move on subculture medium carry out subculture expand it is numerous, wherein, cultivation temperature be 28 DEG C,
Intensity of illumination is 2000lux, and light application time is 12 hours/day, and the subculture medium includes：MS, 3mg/L 6- benzyl glands are fast
Purine, 0.6mg/L gibberellin, 2.0mg/L Methotrexate, 22g/L sucrose and 15g/L agar；
Step 4: after the budlet on the subculture medium is grown to more than 2cm, move it in Rooting and hardening-off culture base
Culture of rootage and strong sprout are carried out, wherein, cultivation temperature is 27 DEG C, intensity of illumination 2000lux, and light application time is 12 hours/day,
The Rooting and hardening-off culture base includes：1/2MS, 5mg/L 6-benzyladenine, 0.6mg/L methyl α-naphthyl acetate, 0.4mg/L it is red
The agar of mycin, 30g/L sucrose and 18g/L；
Step 5: when the tissue-cultured seedling length on the Rooting and hardening-off culture base to 3 leaves, hardening 7 days, to training tissue culture seedling
When growing 5 leaves, you can transplanted.
Wherein, the initial pH value of the inducing culture, subculture medium and Rooting and hardening-off culture base is 6.
In order to illustrate the effect of the cultural method of the common calla tissue-cultured seedling of the present invention, present inventor uses same product
The common calla of kind, A groups are using the common calla tissue-cultured seedling of the method culture of embodiment 1, the common calla that B groups are cultivated using embodiment 2
Tissue-cultured seedling, C groups use the common calla tissue-cultured seedling that embodiment 3 is cultivated, and D groups use the common calla tissue-cultured seedling of outsourcing, are tried in same
Test and plant on the ground, plant means and management method all same, experimental result are shown in Table 1：
The germination percentage and cultivation cycle of the tissue-cultured seedling of table 1
| Group | Tissue-cultured seedling germination percentage/% | Tissue-cultured seedling cultivation cycle/day |
| A | 95.3 | 42 |
| B | 96.1 | 41 |
| C | 95.7 | 45 |
| D | 92.3 | 62 |
As shown in Table 1, using the present invention common calla tissue-cultured seedling cultural method common calla tissue-cultured seedling relative to outsourcing
Common calla tissue-cultured seedling, the germination percentage of tissue-cultured seedling dramatically increases, while the cultivation cycle of tissue-cultured seedling also significantly shortens.
Although embodiment of the present invention is disclosed as above, it is not restricted in specification and embodiment listed
With it can be applied to various suitable the field of the invention completely, can be easily for those skilled in the art
Other modification is realized, therefore under the universal limited without departing substantially from claim and equivalency range, it is of the invention and unlimited
In specific details and shown here as the embodiment with description.
Claims (5)
1. a kind of cultural method of common calla tissue-cultured seedling, it is characterised in that comprise the following steps：
Step 1: it is explant to choose common calla main bud or lateral bud, after use quality fraction is 65% alcohol disinfecting, use is sterile
Water rinses 1-2 minutes, then is placed in the liquor natrii hypochloritis that mass fraction is 15-25% and sterilizes 5 minutes, with aseptic water washing 4-
After 6 minutes, explant is obtained；
Step 2: the explant is placed in into culture in inducing culture extremely forms adventitious bud, wherein, the inducing culture bag
Include：MS, 2-3mg/L 6-benzyladenine, 0.1-0.2mg/L methyl α-naphthyl acetate, 0.2-0.6mg/L gibberellin, 20g/L sucrose
With 12g/L agar；
Step 3: by the adventitious bud move on subculture medium carry out subculture expand it is numerous, wherein, the subculture medium includes：
MS, 2-3mg/L 6-benzyladenine, 0.2-0.6mg/L gibberellin, 0.5-2.0mg/L Methotrexate, 22g/L sucrose
With 15g/L agar；
Step 4: after the budlet on the subculture medium is grown to more than 2cm, move it in Rooting and hardening-off culture base and carry out
Culture of rootage and strong sprout, wherein, the Rooting and hardening-off culture base includes：1/2MS, 3-5mg/L 6-benzyladenine, 0.4-
0.6mg/L methyl α-naphthyl acetate, 0.2-0.4mg/L gibberellin, 30g/L sucrose and 18g/L agar；
Step 5: when the tissue-cultured seedling length on the Rooting and hardening-off culture base to 2-3 piece leaves, hardening 4-7 days, to training tissue culture seedling
When growing 3-5 piece leaves, you can transplanted.
2. the cultural method of common calla tissue-cultured seedling as claimed in claim 1, it is characterised in that culture is not in the step 2
During normal bud, its cultivation temperature is 23-28 DEG C, intensity of illumination 1200-1800lux, and light application time is 8-10 hours/day.
3. the cultural method of common calla tissue-cultured seedling as claimed in claim 1, it is characterised in that subculture is carried out in the step 3
When expanding numerous, its cultivation temperature is 23-28 DEG C, intensity of illumination 1200-2000lux, and light application time is 8-12 hours/day.
4. the cultural method of common calla tissue-cultured seedling as claimed in claim 1, it is characterised in that the step 4 carries out training of taking root
Cultivation temperature during foster and strong sprout is 23-27 DEG C, intensity of illumination 1200-2000lux, and light application time is 8-12 hours/day
Under the conditions of cultivate 15-20 days.
5. the cultural method of common calla tissue-cultured seedling as claimed in claim 1, it is characterised in that the inducing culture, subculture
The initial pH value of culture medium and Rooting and hardening-off culture base is 5-6.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109526751A (en) * | 2018-12-29 | 2019-03-29 | 贵州大学 | A kind of common calla bud eye method for tissue culture |
| CN110972951A (en) * | 2019-12-24 | 2020-04-10 | 云南农业大学 | Method for rejuvenating and breaking dormancy of calla tissue culture seedlings |
| CN112042532A (en) * | 2019-10-18 | 2020-12-08 | 江苏省中国科学院植物研究所 | Culture medium for inducing calluses to generate and application of culture medium in establishment of calluses regeneration system |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104041407A (en) * | 2013-08-08 | 2014-09-17 | 欧中（福建）植物技术有限公司 | Tissue culture rapid propagation of dark purple Calla lily |
-
2017
- 2017-11-14 CN CN201711119709.3A patent/CN107667866A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104041407A (en) * | 2013-08-08 | 2014-09-17 | 欧中（福建）植物技术有限公司 | Tissue culture rapid propagation of dark purple Calla lily |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109526751A (en) * | 2018-12-29 | 2019-03-29 | 贵州大学 | A kind of common calla bud eye method for tissue culture |
| CN112042532A (en) * | 2019-10-18 | 2020-12-08 | 江苏省中国科学院植物研究所 | Culture medium for inducing calluses to generate and application of culture medium in establishment of calluses regeneration system |
| CN112042532B (en) * | 2019-10-18 | 2022-01-11 | 江苏省中国科学院植物研究所 | Culture medium for inducing calluses to generate and application of culture medium in establishment of calluses regeneration system |
| CN110972951A (en) * | 2019-12-24 | 2020-04-10 | 云南农业大学 | Method for rejuvenating and breaking dormancy of calla tissue culture seedlings |
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