CN106106153A - A kind of method for tissue culture of iris - Google Patents
A kind of method for tissue culture of iris Download PDFInfo
- Publication number
- CN106106153A CN106106153A CN201610475650.0A CN201610475650A CN106106153A CN 106106153 A CN106106153 A CN 106106153A CN 201610475650 A CN201610475650 A CN 201610475650A CN 106106153 A CN106106153 A CN 106106153A
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- iris
- tissue culture
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- culture
- germination
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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
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- Life Sciences & Earth Sciences (AREA)
- 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 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.
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 is single stem torrid zone epiphytic orchid, is distributed in Asia, Oceania Perenniporia martius
Area, is grown on the tree dry and wet stone in dark and damp foggy high temperature forest more, and because its flower-shape is unique, pattern is gorgeous, and florescence is long,
Become one of flowers most popular on current flowers market international, domestic, have the laudatory title of " Cymbidium ensifolium (L.) Sw. queen ".Broadcast by aseptic
Plant and tissue culture, it is thus achieved that a large amount of elite plant, fine quality feature can be kept, be by extensive commodity production and new product
Plant the key of selection-breeding.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 by green
Turn Huang；
(2) inoculation
First clean capsule surface with the liquid detergent solution of dilution, then rinse, then soak 10-20min with 8--12%NaClO, so
After on superclean bench, with 75% alcohol disinfecting 1-3min, and sterilize 3min with 0.05-0.15% mercuric chloride, use aseptic water washing
2-4 time, take out capsule and be placed in aseptic culture dish, cut capsule and seed is moved in the culture bottle having sterilized water, shake gently
Move and be 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 after 20 days female
Bottle is cultivated, and 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, described sphagna, coconut palm bran, fragment of brick and Linesless charcoal
Ratio 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) composition of axenic germination culture medium include tryptone, bananas juice, activated carbon and
Agar.
Preferably, the content of described tryptone, bananas juice, activated carbon and agar be 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 basic element of cell division BA with
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, institute
The concentration stating 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, sowing
Density can produce " population effect ", so that the germination of seed speeds, the composition of described axenic germination culture medium includes pancreas egg
White peptone, bananas juice, activated carbon and agar and proportioning can provide the nutrition required for seed growth, promote the sprouting of seed,
While described sprouting add hormone, number of seedling amount can be made to increase, while described sprouting interpolation organic additive, described in have
Machine additive is sucrose, and the concentration of described sucrose is 2-4%, can form iris germination, protocorm and blade life
Long have facilitation.The iris using this kind of method for tissue culture to cultivate out has germination percentage height and grows excellent
Advantage, 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, then soak 10min with 8%NaClO, then ultra-clean
On workbench, with 75% alcohol disinfecting 1min, and with 0.05% mercuric chloride sterilization 3min, with aseptic water washing 2 times, take out capsule and be placed on
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, inoculation
Density be 200/m2；
(3) sprout
Use seed asepsis sprouting culture medium that seed is sprouted, inoculate latter 10 days, embryo germination, after 20 days, proceed to half female bottle training
Supporting, generation, subculture and root culture 3 stage at the beginning of point form band root seedling, and composition and the proportioning of described axenic germination culture medium are
The content of tryptone, bananas juice, activated carbon and agar is respectively 1g/l, 50g/l, 0.5g/l, 8g/l, while described sprouting
Add hormone, 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, adds organic additive while described sprouting, described organic additive is sucrose, and 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, described sphagna, coconut palm bran, fragment of brick and Linesless charcoal
Ratio 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 by green turn
Yellow；
(2) inoculation
First clean capsule surface with the liquid detergent solution of dilution, then rinse, then soak 15min with 10%NaClO, then super
On clean workbench, with 75% alcohol disinfecting 2min, and sterilizing 3min with 0.10% mercuric chloride, with aseptic water washing 3 times, taking-up capsule is put
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, connect
The density planted is 250/m2；
(3) sprout
Use seed asepsis sprouting culture medium that seed is sprouted, inoculate latter 15 days, embryo germination, after 20 days, proceed to half female bottle training
Supporting, generation, subculture and root culture 3 stage at the beginning of point form band root seedling, and composition and the proportioning of described axenic germination culture medium are
The content of tryptone, bananas juice, activated carbon and agar is respectively 2g/l, 100g/l, 1.0g/l, 9g/l, described sprouting same
Shi Tianjia hormone, described hormone consist of the basic element of cell division BA and NAA, the proportioning of described BA Yu NAA be 5mg/l and
0.05mg/l, adds organic additive while described sprouting, described organic additive is sucrose, and 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, described sphagna, coconut palm bran, fragment of brick and Linesless charcoal
Ratio 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 by green turn
Yellow；
(2) inoculation
First clean capsule surface with the liquid detergent solution of dilution, then rinse, then soak 20min with 12%NaClO, then super
On clean workbench, with 75% alcohol disinfecting 3min, and sterilizing 3min with 0.15% mercuric chloride, with aseptic water washing 4 times, taking-up capsule is put
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, connect
The density planted is 300/m2；
(3) sprout
Use seed asepsis sprouting culture medium that seed is sprouted, inoculate latter 1020 days, embryo germination, after 20 days, proceed to half female bottle
Cultivating, generation, subculture and root culture 3 stage at the beginning of point form band root seedling, the composition of described axenic germination culture medium and proportioning
Content for tryptone, bananas juice, activated carbon and agar is respectively 3g/l, 150g/l, 1.5g/l, 10g/l, described sprouting
Add hormone simultaneously, described hormone consist of the basic element of cell division BA and NAA, the proportioning of described BA Yu NAA be 6mg/l and
0.06mg/l, adds organic additive while described sprouting, described organic additive is sucrose, and 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, described sphagna, coconut palm bran, fragment of brick and Linesless charcoal
Ratio 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 than prior art tissue
The seed germination rate of the iris after cultivation is high, and germinating time is short, and leaf growth than prior art tissue culture is fast, now
It is 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 is permissible
Producing " population effect ", so that the germination of seed speeds, the composition of described axenic germination culture medium includes tryptone, Fructus Musae
Juice, activated carbon and agar and proportioning can provide the nutrition required for seed growth, promote the sprouting of seed, described sprouting
Adding hormone simultaneously, number of seedling amount can be made to increase, add organic additive while described sprouting, described organic additive is
Sucrose, the concentration of described sucrose is 2-4%, iris germination, protocorm formation and leaf growth can be had promotion
Effect.The iris using this kind of method for tissue culture to cultivate out has germination percentage height and grows excellent advantage, market
Have a high potential, have a extensive future.
The foregoing is only embodiments of the invention, not thereby limit the scope of the claims of the present invention, every utilize this
Equivalent structure or equivalence flow process that bright description is made convert, or are directly or indirectly used in other relevant technology necks
Territory, is the most in like manner 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 by green
Turn Huang；
(2) inoculation
First clean capsule surface with the liquid detergent solution of dilution, then rinse, then soak 10-20min with 8--12%NaClO, so
After on superclean bench, with 75% alcohol disinfecting 1-3min, and sterilize 3min with 0.05-0.15% mercuric chloride, use aseptic water washing
2-4 time, take out capsule and be placed in aseptic culture dish, cut capsule and seed is moved in the culture bottle having sterilized water, shake gently
Move and be 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 after 20 days female
Bottle is cultivated, and 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, described sphagna, coconut palm bran, fragment of brick and Linesless charcoal
Ratio 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.
2. according to the method for tissue culture of a kind of iris described in claim 1, it is characterised in that: described step connects in (2)
The density planted 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: nothing in described step (3)
The composition of bacterium germination 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: described tryptone, perfume (or spice)
The content of any of several broadleaf plants 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 characterised in that: described step is sprouted in (3)
While Faing 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: described step is sprouted in (3)
Adding organic additive while Faing, 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 |
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| CN201610475650.0A CN106106153A (en) | 2016-06-27 | 2016-06-27 | A kind of method for tissue culture of iris |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610475650.0A CN106106153A (en) | 2016-06-27 | 2016-06-27 | A kind of method for tissue culture of iris |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107047316A (en) * | 2017-06-06 | 2017-08-18 | 郑州市农林科学研究所 | A kind of iris tissue culture method and culture medium |
| CN113826549A (en) * | 2021-09-01 | 2021-12-24 | 海南大学 | Ornamental dendrobium crossbreeding method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102845311A (en) * | 2012-10-11 | 2013-01-02 | 山东鑫秋种业科技有限公司 | Method for preparing phalaenopsis plant tissue medium |
| CN104770294A (en) * | 2015-04-02 | 2015-07-15 | 河南省农业科学院 | Breeding method using protocorm based on germinated phalaenopsis seeds as receptor |
-
2016
- 2016-06-27 CN CN201610475650.0A patent/CN106106153A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102845311A (en) * | 2012-10-11 | 2013-01-02 | 山东鑫秋种业科技有限公司 | Method for preparing phalaenopsis plant tissue medium |
| CN104770294A (en) * | 2015-04-02 | 2015-07-15 | 河南省农业科学院 | Breeding method using protocorm based on germinated phalaenopsis seeds as receptor |
Non-Patent Citations (2)
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| 俞继英等: "蝴蝶兰无菌播种与组织培养研究进展", 《林业科技开发》 * |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107047316A (en) * | 2017-06-06 | 2017-08-18 | 郑州市农林科学研究所 | A kind of iris tissue culture method and culture medium |
| CN107047316B (en) * | 2017-06-06 | 2019-05-24 | 郑州市农林科学研究所 | A kind of iris tissue culture method and culture medium |
| CN113826549A (en) * | 2021-09-01 | 2021-12-24 | 海南大学 | Ornamental dendrobium crossbreeding method |
| CN113826549B (en) * | 2021-09-01 | 2022-06-28 | 海南大学 | Ornamental dendrobium crossbreeding method |
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