CN105941161A - Tissue culture method of phalaenopsis - Google Patents
Tissue culture method of phalaenopsis Download PDFInfo
- Publication number
- CN105941161A CN105941161A CN201610468943.6A CN201610468943A CN105941161A CN 105941161 A CN105941161 A CN 105941161A CN 201610468943 A CN201610468943 A CN 201610468943A CN 105941161 A CN105941161 A CN 105941161A
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- iris
- 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
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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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- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a tissue culture method of phalaenopsis. In the steps, the inoculation density and seeding density can generate a 'group effect' so as to accelerate seed germination; a sterile germination culture medium consists of tryptone, banana juice, active carbon and agar in a ratio and can provide nutrition for the seed growth and promote the seed germination; hormone is added at the time of germination, and the number of live seedlings can be increased; an organic additive is added during germination, wherein the organic additive is 24% sucrose, and a promotion effect can be realized on the phalaenopsis seed germination, protocorm formation and blade growth. By adopting the tissue culture method, the cultured phalaenopsis has the advantages of high germination rate and sound growth and also has huge market potential and broad prospect.
Description
Technical field
The invention belongs to field of flower culture, it is more particularly related to the method for tissue culture of a kind of iris.
Background technology
Iris, the orchid family Phalaenopsis, it it is single stem torrid zone epiphytic orchid, it is distributed in Asia, subtropical and tropical zones, Oceania, is grown on the tree dry and wet stone in dark and damp foggy high temperature forest more, because its flower-shape is unique, pattern is gorgeous, florescence is long, becomes one of flowers most popular on current flowers market international, domestic, have the laudatory title of " Cymbidium ensifolium (L.) Sw. queen ".By aseptic seeding and tissue culture, it is thus achieved that a large amount of elite plant, fine quality feature can be kept, be by the key of extensive commodity production and breeding of new variety.The method for tissue culture of iris also exists slow and that seedling quantity is the highest problem of germinateing at present.
Summary of the invention
Problem to be solved by this invention is to provide the method for tissue culture of a kind of iris.
To achieve these goals, the technical scheme that the present invention takes is:
The method for tissue culture of a kind of iris, comprises the steps:
(1) collect seed
First choose after pollination 95-105 days iris fruit pod, selected fruit pod is that outer smooth is full, and color is from green to yellow；
(2) inoculation
First clean capsule surface with the liquid detergent solution of dilution, then rinse, 10-20min is soaked again with 8--12%NaClO, then on superclean bench, with 75% alcohol disinfecting 1-3min, and sterilize 3min with 0.05-0.15% mercuric chloride, with aseptic water washing 2-4 time, take out capsule to be placed in aseptic culture dish, cut capsule and seed is moved in the culture bottle having sterilized water, be shaken gently for being allowed to be uniformly dispersed；
(3) sprout
Use seed asepsis sprouting culture medium that seed is sprouted, after inoculation 10-20 days, embryo germination, proceed to half female bottle after 20 days and cultivate, generation, subculture and root culture 3 stage at the beginning of point form band root seedling；
(4) seedling exercising and transplanting
Can transplant by bottle outlet when 3-5 sheet leaf, 2-4 bar root occurs in seedling；
(5) selection of medium is transplanted
The mixture that medium is sphagna, coconut palm bran, fragment of brick and Linesless charcoal transplanted is substrate, and the ratio of described sphagna, coconut palm bran, fragment of brick and Linesless charcoal is 0.5:0.5:0.3:1-1:1:0.5:2；
(6) temperature treatment
Temperature in sowing time, nursery stage controls at 20-24 DEG C, and the temperature of Seedling Stage controls at 16-18 DEG C.
Preferably, in described step (2), the density of inoculation is 200-300 grain/m2。
Preferably, in described step (3), the composition of axenic germination culture medium includes tryptone, bananas juice, activated carbon and agar.
Preferably, the content of described tryptone, bananas juice, activated carbon and agar is respectively 1-3g/l, 50-150g/l, 0.5-1.5g/l, 8-10g/l.
Preferably, while described step (3) is sprouted, add hormone, described hormone consist of the basic element of cell division BA and NAA, the proportioning of described BA Yu NAA is 4-6mg/l and 0.04-0.06mg/l.
Preferably, adding organic additive in described step (3) while sprouting, described organic additive is sucrose, and the concentration of described sucrose is 2-4%.
Beneficial effect: the invention provides the method for tissue culture of a kind of iris, the density of described step inoculation, thickness of sowing can produce " population effect ", so that the germination of seed speeds, the composition of described axenic germination culture medium includes tryptone, bananas juice, activated carbon and agar and proportioning can provide the nutrition required for seed growth, promote the sprouting of seed, hormone is added while described sprouting, number of seedling amount can be made to increase, organic additive is added while described sprouting, described organic additive is sucrose, the concentration of described sucrose is 2-4%, can be to iris germination, protocorm is formed and leaf growth has facilitation.The iris using this kind of method for tissue culture to cultivate out has germination percentage height and grows excellent advantage, and market potential is huge, has a extensive future.
Detailed description of the invention
Embodiment
1
:
The method for tissue culture of a kind of iris, comprises the steps:
(1) collect seed
First choose pollination latter 95 days iris fruit pod, selected fruit pod is that outer smooth is full, and color is from green to yellow；
(2) inoculation
First clean capsule surface with the liquid detergent solution of dilution, then rinse, 10min is soaked again, then on superclean bench, with 75% alcohol disinfecting 1min with 8%NaClO, and with 0.05% mercuric chloride sterilization 3min, with aseptic water washing 2 times, take out capsule and be placed in aseptic culture dish, cut capsule and seed is moved in the culture bottle having sterilized water, being shaken gently for being allowed to be uniformly dispersed, the density of inoculation is 200/m2；
(3) sprout
Use seed asepsis sprouting culture medium that seed is sprouted, inoculate latter 10 days, embryo germination, proceed to half female bottle after 20 days cultivate, generation at the beginning of Fen, subculture and root culture 3 stage form band root seedling, the composition of described axenic germination culture medium and proportioning are tryptone, bananas juice, the content of activated carbon and agar is respectively 1g/l, 50g/l, 0.5g/l, 8g/l, hormone is added while described sprouting, described hormone consist of the basic element of cell division BA and NAA, the proportioning of described BA Yu NAA is 4mg/l and 0.04mg/l, organic additive is added while described sprouting, described organic additive is sucrose, the concentration of described sucrose is 2%；
(4) seedling exercising and transplanting
Can transplant by bottle outlet when 3 leaves, 2 roots occurs in seedling；
(5) selection of medium is transplanted
The mixture that medium is sphagna, coconut palm bran, fragment of brick and Linesless charcoal transplanted is substrate, and the ratio of described sphagna, coconut palm bran, fragment of brick and Linesless charcoal is 0.5:0.5:0.3:1；
(6) temperature treatment
Temperature in sowing time, nursery stage controls at 20 DEG C, and the temperature of Seedling Stage controls at 16 DEG C.
Embodiment
2
:
The method for tissue culture of a kind of iris, comprises the steps:
(1) collect seed
First choose pollination latter 100 days iris fruit pod, selected fruit pod is that outer smooth is full, and color is from green to yellow；
(2) inoculation
First clean capsule surface with the liquid detergent solution of dilution, then rinse, 15min is soaked again, then on superclean bench, with 75% alcohol disinfecting 2min with 10%NaClO, and with 0.10% mercuric chloride sterilization 3min, with aseptic water washing 3 times, take out capsule and be placed in aseptic culture dish, cut capsule and seed is moved in the culture bottle having sterilized water, being shaken gently for being allowed to be uniformly dispersed, the density of inoculation is 250/m2；
(3) sprout
Use seed asepsis sprouting culture medium that seed is sprouted, inoculate latter 15 days, embryo germination, proceed to half female bottle after 20 days cultivate, generation at the beginning of Fen, subculture and root culture 3 stage form band root seedling, the composition of described axenic germination culture medium and proportioning are tryptone, bananas juice, the content of activated carbon and agar is respectively 2g/l, 100g/l, 1.0g/l, 9g/l, hormone is added while described sprouting, described hormone consist of the basic element of cell division BA and NAA, the proportioning of described BA Yu NAA is 5mg/l and 0.05mg/l, organic additive is added while described sprouting, described organic additive is sucrose, the concentration of described sucrose is 3%；
(4) seedling exercising and transplanting
Can transplant by bottle outlet when 4 leaves, 3 roots occurs in seedling；
(5) selection of medium is transplanted
The mixture that medium is sphagna, coconut palm bran, fragment of brick and Linesless charcoal transplanted is substrate, and the ratio of described sphagna, coconut palm bran, fragment of brick and Linesless charcoal is 0.75:0.75:0.4:1.5；
(6) temperature treatment
Temperature in sowing time, nursery stage controls at 22 DEG C, and the temperature of Seedling Stage controls at 18 DEG C.
Embodiment
3
:
The method for tissue culture of a kind of iris, comprises the steps:
(1) collect seed
First choose pollination latter 105 days iris fruit pod, selected fruit pod is that outer smooth is full, and color is from green to yellow；
(2) inoculation
First clean capsule surface with the liquid detergent solution of dilution, then rinse, 20min is soaked again, then on superclean bench, with 75% alcohol disinfecting 3min with 12%NaClO, and with 0.15% mercuric chloride sterilization 3min, with aseptic water washing 4 times, take out capsule and be placed in aseptic culture dish, cut capsule and seed is moved in the culture bottle having sterilized water, being shaken gently for being allowed to be uniformly dispersed, the density of inoculation is 300/m2；
(3) sprout
Use seed asepsis sprouting culture medium that seed is sprouted, inoculate latter 1020 days, embryo germination, proceed to half female bottle after 20 days cultivate, generation at the beginning of Fen, subculture and root culture 3 stage form band root seedling, the composition of described axenic germination culture medium and proportioning are tryptone, bananas juice, the content of activated carbon and agar is respectively 3g/l, 150g/l, 1.5g/l, 10g/l, hormone is added while described sprouting, described hormone consist of the basic element of cell division BA and NAA, the proportioning of described BA Yu NAA is 6mg/l and 0.06mg/l, organic additive is added while described sprouting, described organic additive is sucrose, the concentration of described sucrose is 4%；
(4) seedling exercising and transplanting
Can transplant by bottle outlet when 5 leaves, 4 roots occurs in seedling；
(5) selection of medium is transplanted
The mixture that medium is sphagna, coconut palm bran, fragment of brick and Linesless charcoal transplanted is substrate, and the ratio of described sphagna, coconut palm bran, fragment of brick and Linesless charcoal is 1:1:0.5:2；
(6) temperature treatment
Temperature in sowing time, nursery stage controls at 24 DEG C, and the temperature of Seedling Stage controls at 18 DEG C.
After above method, taking out sample respectively, measurement result is as follows:
| Detection project | Embodiment 1 | Embodiment 2 | Embodiment 3 | Existing index |
| Germinate (d) | 3 | 1 | 2 | 5 |
| Seed germination rate (%) | 97 | 99 | 96 | 90 |
| Leaf growth | Hurry up | Hurry up | Hurry up | Comparatively fast |
Can draw according to above table data, when embodiment 2 parameter, the iris after tissue culture is higher than the seed germination rate of the iris after prior art tissue culture, and germinating time is short, leaf growth than prior art tissue culture is fast, is now more beneficial for the tissue culture of iris.
The invention provides the method for tissue culture of a kind of iris, the density of described step inoculation, thickness of sowing can produce " population effect ", so that the germination of seed speeds, the composition of described axenic germination culture medium includes tryptone, bananas juice, activated carbon and agar and proportioning can provide the nutrition required for seed growth, promote the sprouting of seed, hormone is added while described sprouting, number of seedling amount can be made to increase, organic additive is added while described sprouting, described organic additive is sucrose, the concentration of described sucrose is 2-4%, can be to iris germination, protocorm is formed and leaf growth has facilitation.The iris using this kind of method for tissue culture to cultivate out has germination percentage height and grows excellent advantage, and market potential is huge, has a extensive future.
The foregoing is only embodiments of the invention; not thereby the scope of the claims of the present invention is limited; every equivalent structure utilizing description of the invention content to be made or equivalence flow process conversion; or directly or indirectly it is used in other relevant technical fields, the most in like manner it is included in the scope of patent protection of the present invention.
Claims (6)
1. the method for tissue culture of an iris, it is characterised in that comprise the steps:
(1) collect seed
First choose after pollination 95-105 days iris fruit pod, selected fruit pod is that outer smooth is full, and color is from green to yellow；
(2) inoculation
First clean capsule surface with the liquid detergent solution of dilution, then rinse, 10-20min is soaked again with 8--12%NaClO, then on superclean bench, with 75% alcohol disinfecting 1-3min, and sterilize 3min with 0.05-0.15% mercuric chloride, with aseptic water washing 2-4 time, take out capsule to be placed in aseptic culture dish, cut capsule and seed is moved in the culture bottle having sterilized water, be shaken gently for being allowed to be uniformly dispersed；
(3) sprout
Use seed asepsis sprouting culture medium that seed is sprouted, after inoculation 10-20 days, embryo germination, proceed to half female bottle after 20 days and cultivate, generation, subculture and root culture 3 stage at the beginning of point form band root seedling；
(4) seedling exercising and transplanting
Can transplant by bottle outlet when 3-5 sheet leaf, 2-4 bar root occurs in seedling
；
(5) selection of medium is transplanted
The mixture that medium is sphagna, coconut palm bran, fragment of brick and Linesless charcoal transplanted is substrate, and the ratio of described sphagna, coconut palm bran, fragment of brick and Linesless charcoal is 0.5:0.5:0.3:1-1:1:0.5:2；
(6) temperature treatment
Temperature in sowing time, nursery stage controls
20-24 DEG C, the temperature of Seedling Stage controls
16-18 ℃。
2. according to the method for tissue culture of a kind of iris described in claim 1, it is characterised in that: in described step (2), the density of inoculation is 200-300 grain/m2。
3. according to the method for tissue culture of a kind of iris described in claim 1, it is characterised in that: in described step (3), the composition of axenic germination culture medium includes tryptone, bananas juice, activated carbon and agar.
4. according to the method for tissue culture of a kind of iris described in claim 3, it is characterised in that: the content of described tryptone, bananas juice, activated carbon and agar is respectively 1-3g/l, 50-150g/l, 0.5-1.5g/l, 8-10g/l.
5. according to the method for tissue culture of a kind of iris described in claim 1, it is characterized in that: while described step (3) is sprouted, add hormone, described hormone consist of the basic element of cell division BA and NAA, the proportioning of described BA Yu NAA is 4-6mg/l and 0.04-0.06mg/l.
6. according to the method for tissue culture of a kind of iris described in claim 1, it is characterised in that: adding organic additive while sprouting in described step (3), described organic additive is sucrose, and the concentration of described sucrose is 2-4%.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610468943.6A CN105941161A (en) | 2016-06-25 | 2016-06-25 | Tissue culture method of phalaenopsis |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610468943.6A CN105941161A (en) | 2016-06-25 | 2016-06-25 | Tissue culture method of phalaenopsis |
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| CN201610468943.6A Withdrawn CN105941161A (en) | 2016-06-25 | 2016-06-25 | Tissue culture method of phalaenopsis |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108496753A (en) * | 2018-03-07 | 2018-09-07 | 山东省烟台市农业科学研究院 | A method of delaying iris cultivation matrix acidification corruption |
| CN112425509A (en) * | 2020-12-17 | 2021-03-02 | 青州市亚泰农业科技有限公司 | Sterile culture medium for waxy phalaenopsis industrial tissue culture |
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| CN102283110A (en) * | 2010-09-29 | 2011-12-21 | 深圳市农科植物克隆种苗有限公司 | Method for rapidly propagating moth orchids through inducing cluster buds by leaves |
| CN102613076A (en) * | 2012-03-26 | 2012-08-01 | 吴海红 | Vegetative propagation method for butterfly orchid |
| CN103988780A (en) * | 2014-05-23 | 2014-08-20 | 中国长江三峡集团公司 | Method for directly inducing adventitious buds of butterfly orchid |
| CN104686362A (en) * | 2015-03-24 | 2015-06-10 | 芜湖东源新农村开发股份有限公司 | Butterfly orchid tissue culturing method |
| CN105918134A (en) * | 2016-06-15 | 2016-09-07 | 安徽菲扬农业科技有限公司 | Tissue culture method of Phalaenopsis |
-
2016
- 2016-06-25 CN CN201610468943.6A patent/CN105941161A/en not_active Withdrawn
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102283110A (en) * | 2010-09-29 | 2011-12-21 | 深圳市农科植物克隆种苗有限公司 | Method for rapidly propagating moth orchids through inducing cluster buds by leaves |
| CN102613076A (en) * | 2012-03-26 | 2012-08-01 | 吴海红 | Vegetative propagation method for butterfly orchid |
| CN103988780A (en) * | 2014-05-23 | 2014-08-20 | 中国长江三峡集团公司 | Method for directly inducing adventitious buds of butterfly orchid |
| CN104686362A (en) * | 2015-03-24 | 2015-06-10 | 芜湖东源新农村开发股份有限公司 | Butterfly orchid tissue culturing method |
| CN105918134A (en) * | 2016-06-15 | 2016-09-07 | 安徽菲扬农业科技有限公司 | Tissue culture method of Phalaenopsis |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108496753A (en) * | 2018-03-07 | 2018-09-07 | 山东省烟台市农业科学研究院 | A method of delaying iris cultivation matrix acidification corruption |
| CN112425509A (en) * | 2020-12-17 | 2021-03-02 | 青州市亚泰农业科技有限公司 | Sterile culture medium for waxy phalaenopsis industrial tissue culture |
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