CN101869066A - Tissue culture method of African agapanthus - Google Patents
Tissue culture method of African agapanthus Download PDFInfo
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- CN101869066A CN101869066A CN200910049974A CN200910049974A CN101869066A CN 101869066 A CN101869066 A CN 101869066A CN 200910049974 A CN200910049974 A CN 200910049974A CN 200910049974 A CN200910049974 A CN 200910049974A CN 101869066 A CN101869066 A CN 101869066A
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- 244000045257 Agapanthus umbellatus Species 0.000 title abstract 3
- 238000012136 culture method Methods 0.000 title abstract 2
- 239000000463 material Substances 0.000 claims abstract description 7
- 230000004069 differentiation Effects 0.000 claims abstract description 6
- 241000499929 Agapanthus Species 0.000 claims description 24
- 238000000034 method Methods 0.000 claims description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 15
- 241000196324 Embryophyta Species 0.000 claims description 14
- 239000001963 growth medium Substances 0.000 claims description 14
- 230000001939 inductive effect Effects 0.000 claims description 14
- 229920001817 Agar Polymers 0.000 claims description 10
- 239000008272 agar Substances 0.000 claims description 10
- 238000007654 immersion Methods 0.000 claims description 8
- 239000002609 medium Substances 0.000 claims description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 5
- 206010020649 Hyperkeratosis Diseases 0.000 claims description 5
- 241000283986 Lepus Species 0.000 claims description 5
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 5
- 229930006000 Sucrose Natural products 0.000 claims description 5
- 238000011010 flushing procedure Methods 0.000 claims description 5
- 238000005286 illumination Methods 0.000 claims description 5
- 230000006698 induction Effects 0.000 claims description 5
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 claims description 5
- 229910052753 mercury Inorganic materials 0.000 claims description 5
- 230000000442 meristematic effect Effects 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 5
- 239000005720 sucrose Substances 0.000 claims description 5
- 230000004083 survival effect Effects 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 5
- 238000010923 batch production Methods 0.000 abstract description 2
- 238000005516 engineering process Methods 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 1
- 206010020880 Hypertrophy Diseases 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- -1 iginates Species 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
Abstract
The invention relates to a tissue culture method of African agapanthus, comprising the following steps of: acquisition of a sterile material, differentiation and propagation of corms, strong seedling culture of corms, rooting culture, seedling hardening, transplanting, and the like. Compared with the prior art, the invention greatly improves the propagation speed of the African agapanthus and the neatness of seedlings and can realize industrial large-batch production of the seedlings.
Description
Technical field
The present invention relates to the method for tissue culture of a plant species, especially relate to the method for tissue culture of a kind of Afriocan agapanthus.
Background technology
Afriocan agapanthus is an Amaryllidaceae Agapanthus plant, originates in a Peru and a Brazilian band, and existing cultivation is comparatively extensive.Afriocan agapanthus is a herbaceos perennial, and the bulb hypertrophy is subsphaeroidal, and diameter 5-7cm, crust are light green or yellowish-brown.The blade both sides are to life, band shape, and tip is point gradually, and 6-8 piece altogether, blade bears more than after spending.The top is blossomed 2-4, and flower is very large, and the pattern purple is good presbyopic glasses and garden material.As the new varieties of external introduction, Afriocan agapanthus introduce a fine variety negligible amounts, it is slow to plant the ball plant division, the seedling supply is restricted.
Summary of the invention
Purpose of the present invention is exactly the method for tissue culture that a kind of Afriocan agapanthus of the regularity that improves reproduction speed and seedling is provided in order to overcome the defective that above-mentioned prior art exists.
Purpose of the present invention can be achieved through the following technical solutions:
The method for tissue culture of Afriocan agapanthus is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
When Afriocan agapanthus blooms, win the petal of not blooming, with behind the running water flushing 1-3h on ultra-clean work team workbench, utilize the alcohol immersion 10-50s of mass concentration successively for 60-75%, volumetric concentration is that the mercury of 0.5-2 ‰ soaks 10-30min, use aseptic water washing 4-6 time again, utilize aseptic filter paper to blot the moisture on petal surface after, petal is inoculated on the petal inducing culture;
(2) differentiation of bulb and propagation
Petal is inoculated on the petal inducing culture, 1-3 after week petal begin to expand, white projection appears, 3-5 is visible bud meristematic tissue after week, cultivates 1-2 month again, has bottom set to generate, to carry out enrichment culture in the bottom set cutting-out immigration bulb proliferated culture medium, the callus of bulb base portion is more, but bulb grows well bad phenomenon such as no vitrifying;
(3) bulb strong seedling culture
The bulb that induces on the bulb proliferated culture medium gone on the strong seedling culture base grow, can grow up to the bulb of 0.2-0.5cm after 20-40 days;
(4) culture of rootage
Get the bulb of 0.2-0.5cm, root induction in the root media is gone in switching, and the seedling base section dissolves the root original hase of white after 10-20 days, can grow to 2-4cm after 30-35 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 90-100%;
(5) hardening and transplanting
Culture of rootage 30-35 days, when root system grows to 0.5-1cm, select the aseptic seedling of well developed root system, robust growth, in indoor uncork hardening 1-3 days, then seedling is taken out, clean the agar of root, plant in the seedbed in greenhouse and tamed 30-50 days, can transplant outdoor go up basin or plant, give rich water quality management, final transplanting survival rate is 90-100%.
Described petal inducing culture comprises MS+6-BA1.0-5.0mg/L+NAA0.1-0.5mg/L.
The preferred MS+6-BA5.0mg/L+NAA0.5mg/L of described petal inducing culture.
Described bulb proliferated culture medium comprises MS+6-BA0.5-4.0mg/L+NAA0.1-0.3mg/L.
The preferred MS+6-BA2.0mg/L+NAA0.2mg/L of described bulb proliferated culture medium.
Described strong seedling culture base comprises MS+6-BA0.5-2.0mg/L+NAA0.1-0.3mg/L.
The preferred MS+6-BA1.0mg/L+NAA0.1mg/L of described strong seedling culture base.
Described root media comprises MS+NAA0.1-0.3mg/L.
The preferred MS+NAA0.1mg/L of described root media.
Described medium also comprises sucrose 20-40g/L, agar powder 4-8g/L, medium pH 5.5-6.0, cultivation temperature 24-26 ℃, illumination 1500-2500lx.
Compared with prior art, the present invention has greatly improved the reproduction speed of Afriocan agapanthus and the regularity of seedling by tissue culture technology, can realize the batch production production in enormous quantities of growing seedlings.
Embodiment
The present invention is described in detail below in conjunction with specific embodiment.
Embodiment 1
(1) acquisition of sterilizable material
When Afriocan agapanthus blooms, win the petal of not blooming, with behind the running water flushing 1h on ultra-clean work team workbench, utilizing mass concentration successively is 60% alcohol immersion 10s, volumetric concentration is 0.5 ‰ mercury immersion 10min, aseptic water washing 4 times, blot surface moisture with aseptic filter paper after, petal is inoculated on the axillalry bud inducing culture that comprises MS+6-BA1.0mg/L+NAA0.1mg/L;
(2) differentiation of bulb and propagation
Petal is inoculated on the petal inducing culture, petal begins to expand after 1 week, white projection appears, the visible bud meristematic tissue in 3 week backs was cultivated 1 month again, had bottom set to generate, bottom set downcut moved in the bulb proliferated culture medium that comprises MS+6-BA0.5mg/L+NAA0.1mg/L carry out enrichment culture, the callus of bulb base portion is more, but bulb grows well bad phenomenon such as no vitrifying;
(3) bulb strong seedling culture
To go on the strong seedling culture base that comprises MS+6-BA0.5mg/L+NAA0.1mg/L at the bulb that induces on the bulb proliferated culture medium and grow, can grow up to the bulb of 0.2cm after 20 days;
(4) culture of rootage
Get the bulb of 0.2cm, root induction in the root media of going into to comprise MS+NAA0.1mg/L of transferring, the seedling base section dissolves the root original hase of white after 10 days, can grow to 2cm after 30 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 90%;
(5) hardening and transplanting
Culture of rootage 30 days, when root system grows to 0.5cm, select the aseptic seedling of well developed root system, robust growth, in indoor uncork hardening 1 day, then seedling is taken out, clean the agar of root, plant in the seedbed in greenhouse and tamed 30 days, can transplant outdoor go up basin or plant, give rich water quality management, final transplanting survival rate is 90%.
The medium of above-mentioned various situations also comprises sucrose 20g/L, agar powder 4g/L, pH=5.5,24 ℃ of cultivation temperature, illumination 1500lx.
Embodiment 2
(1) acquisition of sterilizable material
When Afriocan agapanthus blooms, win the petal of not blooming, with behind the running water flushing 1h on ultra-clean work team workbench, utilizing mass concentration successively is 75% alcohol immersion 30s, volumetric concentration is 1 ‰ mercury immersion 15min, aseptic water washing 5 times, blot surface moisture with aseptic filter paper after, petal is inoculated on the axillalry bud inducing culture that comprises MS+6-BA5.0mg/L+NAA0.5mg/L;
(2) differentiation of bulb and propagation
Petal is inoculated on the petal inducing culture, petal begins to expand after 3 weeks, white projection appears, the visible bud meristematic tissue in 4 week backs is cultivated 1 first quarter moon again, has bottom set to generate, bottom set downcut moved in the bulb proliferated culture medium that comprises MS+6-BA2.0mg/L+NAA0.2mg/L carry out enrichment culture, the callus of bulb base portion is more, but bulb grows well bad phenomenon such as no vitrifying;
(3) bulb strong seedling culture
To go on the strong seedling culture base that comprises MS+6-BA1.0mg/L+NAA0.1mg/L at the bulb that induces on the bulb proliferated culture medium and grow, can grow up to the bulb of 0.5cm after 30 days;
(4) culture of rootage
Get the bulb of 0.5cm, root induction in the root media of going into to comprise MS+NAA0.1mg/L of transferring, the seedling base section dissolves the root original hase of white after 15 days, can grow to 4cm after 30 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 100%;
(5) hardening and transplanting
Culture of rootage 30 days, when root system grows to 1cm, select the aseptic seedling of well developed root system, robust growth, in indoor uncork hardening 3 days, then seedling is taken out, clean the agar of root, plant in the seedbed in greenhouse and tamed 40 days, can transplant outdoor go up basin or plant, give rich water quality management, final transplanting survival rate is 100%.
The medium of above-mentioned various situations also comprises sucrose 30g/L, agar powder 6g/L, pH=5.8,25 ℃ of cultivation temperature, illumination 2000lx.
Embodiment 3
(1) acquisition of sterilizable material
When Afriocan agapanthus blooms, win the petal of not blooming, with behind the running water flushing 3h on ultra-clean work team workbench, utilizing mass concentration successively is 75% alcohol immersion 50s, volumetric concentration is 2 ‰ mercury immersion 30min, aseptic water washing 6 times, blot surface moisture with aseptic filter paper after, petal is inoculated on the axillalry bud inducing culture that comprises MS+6-BA5.0mg/L+NAA0.5mg/L;
(2) differentiation of bulb and propagation
Petal is inoculated on the petal inducing culture, petal begins to expand after 3 weeks, white projection appears, the visible bud meristematic tissue in 5 week backs was cultivated 2 months again, had bottom set to generate, bottom set downcut moved in the bulb proliferated culture medium that comprises MS+6-BA4.0mg/L+NAA0.3mg/L carry out enrichment culture, the callus of bulb base portion is more, but bulb grows well bad phenomenon such as no vitrifying;
(3) bulb strong seedling culture
To go on the strong seedling culture base that comprises MS+6-BA2.0mg/L+NAA0.2mg/L at the bulb that induces on the bulb proliferated culture medium and grow, can grow up to the bulb of 0.4cm after 40 days;
(4) culture of rootage
Get the bulb of 0.4cm, root induction in the root media of going into to comprise MS+NAA0.3mg/L of transferring, the seedling base section dissolves the root original hase of white after 20 days, can grow to 3cm after 35 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 95%;
(5) hardening and transplanting
Culture of rootage 35 days, when root system grows to 0.8cm, select the aseptic seedling of well developed root system, robust growth, in indoor uncork hardening 3 days, then seedling is taken out, clean the agar of root, plant in the seedbed in greenhouse and tamed 50 days, can transplant outdoor go up basin or plant, give rich water quality management, final transplanting survival rate is 94%.
The medium of above-mentioned various situations also comprises sucrose 40g/L, agar powder 8g/L, pH=6.0,26 ℃ of cultivation temperature, illumination 2500lx.
Claims (10)
1. the method for tissue culture of Afriocan agapanthus is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
When Afriocan agapanthus blooms, win the petal of not blooming, with behind the running water flushing 1-3h on ultra-clean work team workbench, utilize the alcohol immersion 10-50s of mass concentration successively for 60-75%, volumetric concentration is that the mercury of 0.5-2 ‰ soaks 10-30min, use aseptic water washing 4-6 time again, utilize aseptic filter paper to blot the moisture on petal surface after, petal is inoculated on the petal inducing culture;
(2) differentiation of bulb and propagation
Petal is inoculated on the petal inducing culture, 1-3 after week petal begin to expand, white projection appears, 3-5 is visible bud meristematic tissue after week, cultivates 1-2 month again, has bottom set to generate, to carry out enrichment culture in the bottom set cutting-out immigration bulb proliferated culture medium, the callus of bulb base portion is more, but bulb grows well bad phenomenon such as no vitrifying;
(3) bulb strong seedling culture
The bulb that induces on the bulb proliferated culture medium gone on the strong seedling culture base grow, can grow up to the bulb of 0.2-0.5cm after 20-40 days;
(4) culture of rootage
Get the bulb of 0.2-0.5cm, root induction in the root media is gone in switching, and the seedling base section dissolves the root original hase of white after 10-20 days, can grow to 2-4cm after 30-35 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 90-100%;
(5) hardening and transplanting
Culture of rootage 30-35 days, when root system grows to 0.5-1cm, select the aseptic seedling of well developed root system, robust growth, in indoor uncork hardening 1-3 days, then seedling is taken out, clean the agar of root, plant in the seedbed in greenhouse and tamed 30-50 days, can transplant outdoor go up basin or plant, give rich water quality management, final transplanting survival rate is 90-100%.
2. the method for tissue culture of Afriocan agapanthus according to claim 1 is characterized in that, described petal inducing culture comprises MS+6-BA1.0-5.0mg/L+NAA0.1-0.5mg/L.
3. the method for tissue culture of Afriocan agapanthus according to claim 2 is characterized in that, the preferred MS+6-BA5.0mg/L+NAA0.5mg/L of described petal inducing culture.
4. the method for tissue culture of Afriocan agapanthus according to claim 1 is characterized in that, described bulb proliferated culture medium comprises MS+6-BA0.5-4.0mg/L+NAA0.1-0.3mg/L.
5. the method for tissue culture of Afriocan agapanthus according to claim 4 is characterized in that, the preferred MS+6-BA2.0mg/L+NAA0.2mg/L of described bulb proliferated culture medium.
6. the method for tissue culture of Afriocan agapanthus according to claim 1 is characterized in that, described strong seedling culture base comprises MS+6-BA0.5-2.0mg/L+NAA0.1-0.3mg/L.
7. the method for tissue culture of Afriocan agapanthus according to claim 6 is characterized in that, the preferred MS+6-BA1.0mg/L+NAA0.1mg/L of described strong seedling culture base.
8. the method for tissue culture of Afriocan agapanthus according to claim 1 is characterized in that, described root media comprises MS+NAA0.1-0.3mg/L.
9. the method for tissue culture of Afriocan agapanthus according to claim 8 is characterized in that, the preferred MS+NAA0.1mg/L of described root media.
10. according to the method for tissue culture of claim 2 or 4 or 6 or 8 described Afriocan agapanthus, it is characterized in that described medium also comprises sucrose 20-40g/L, agar powder 4-8g/L, medium pH 5.5-6.0, cultivation temperature 24-26 ℃, illumination 1500-2500lx.
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| CN2009100499748A CN101869066B (en) | 2009-04-24 | 2009-04-24 | Tissue culture method of African agapanthus |
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| CN2009100499748A CN101869066B (en) | 2009-04-24 | 2009-04-24 | Tissue culture method of African agapanthus |
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| CN101869066A true CN101869066A (en) | 2010-10-27 |
| CN101869066B CN101869066B (en) | 2012-01-04 |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102823582A (en) * | 2012-09-18 | 2012-12-19 | 上海交通大学 | Vitrification ultralow-temperature preserving method for agapanthus embryogenic callus |
| CN103947548A (en) * | 2014-04-11 | 2014-07-30 | 宁波城市职业技术学院 | Method for establishing agapanthus high-frequency regeneration system |
| CN104170818A (en) * | 2014-09-15 | 2014-12-03 | 上海交通大学 | Method for optimizing vitrified cryopreservation effect of agapanthus embryonic calluses |
| CN107155883A (en) * | 2017-05-05 | 2017-09-15 | 江苏东郁园林科技有限公司 | A kind of Afriocan agapanthus tissue culture of sprout mating system |
| CN109220791A (en) * | 2018-09-12 | 2019-01-18 | 上海市农业科学院 | A kind of tissue culture method using bulb breeding Hipeastrum vittalum (L Her.) Herb.- Amaryllisvittata Ait |
| CN114532227A (en) * | 2022-03-07 | 2022-05-27 | 上海交通大学 | Method for inducing and proliferating calluses of agapanthus radicis roots and tips |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1961654A (en) * | 2006-11-21 | 2007-05-16 | 上海光兆植物速生技术有限公司 | Agipanthus tissue cultivation quick-propagation method |
| CN101129131B (en) * | 2007-09-27 | 2011-03-23 | 上海交通大学 | Non-asepsis seedling re-cutting seedling-producing method of hippeastrum |
-
2009
- 2009-04-24 CN CN2009100499748A patent/CN101869066B/en not_active Expired - Fee Related
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102823582A (en) * | 2012-09-18 | 2012-12-19 | 上海交通大学 | Vitrification ultralow-temperature preserving method for agapanthus embryogenic callus |
| CN103947548A (en) * | 2014-04-11 | 2014-07-30 | 宁波城市职业技术学院 | Method for establishing agapanthus high-frequency regeneration system |
| CN104170818A (en) * | 2014-09-15 | 2014-12-03 | 上海交通大学 | Method for optimizing vitrified cryopreservation effect of agapanthus embryonic calluses |
| CN104170818B (en) * | 2014-09-15 | 2015-08-26 | 上海交通大学 | A kind of method optimizing Agipanthus embryo callus vitrification ultra-low temperature preservation effect |
| CN107155883A (en) * | 2017-05-05 | 2017-09-15 | 江苏东郁园林科技有限公司 | A kind of Afriocan agapanthus tissue culture of sprout mating system |
| CN107155883B (en) * | 2017-05-05 | 2019-06-11 | 江苏东郁植物科技有限公司 | A kind of Afriocan agapanthus tissue culture of sprout mating system |
| CN109220791A (en) * | 2018-09-12 | 2019-01-18 | 上海市农业科学院 | A kind of tissue culture method using bulb breeding Hipeastrum vittalum (L Her.) Herb.- Amaryllisvittata Ait |
| CN109220791B (en) * | 2018-09-12 | 2021-10-08 | 上海市农业科学院 | Tissue culture method for breeding hippeastrum rutilum by using bulbs |
| CN114532227A (en) * | 2022-03-07 | 2022-05-27 | 上海交通大学 | Method for inducing and proliferating calluses of agapanthus radicis roots and tips |
| CN114532227B (en) * | 2022-03-07 | 2022-11-11 | 上海交通大学 | Method for inducing and proliferating calluses of agapanthus radicis roots and tips |
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| CN101869066B (en) | 2012-01-04 |
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Effective date of registration: 20140103 Address after: 201114 Pujiang village, Pujiang Town, Shanghai, Minhang District Patentee after: Shanghai Shangfang Landscape Plant Institute Patentee after: State Grid Shanghai Municipal Electric Power Company Patentee after: Shanghai Urban Power Supply Design Co., Ltd. Address before: 201114 Pujiang village, Pujiang Town, Shanghai, Minhang District Patentee before: Shanghai Shangfang Landscape Plant Institute |
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Correction item: Patentee Correct: Shanghai Main Garden Plant Research Institute Co., Ltd.|201114. Pujiang village, Pujiang Town, Shanghai, Minhang District|Shanghai Shanghai Municipal Electric Power Company power supply design Co., Ltd. False: Shanghai Main Garden Plant Research Institute|201114. Pujiang village, Pujiang Town, Shanghai, Minhang District|Shanghai Shanghai Municipal Electric Power Company power supply design Co., Ltd. Number: 05 Volume: 30 |
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Free format text: CORRECT: PATENTEE; FROM: SHANGHAI SHANGFANG LANDSCAPE PLANT INSTITUTE:201114 MINHANG, SHANGHAI; STATE GRID SHANGHAI ELECTRIC POWER COMPANY, POWER SUPPLY DESIGN (SHANGHAI) CO., LTD. TO: SHANGHAI SHANGFANG LANDSCAPE PLANT RESEARCH INSTITUTE CO., LTD.:201114 MINHANG, SHANGHAI; STATE GRID SHANGHAI ELECTRIC POWER COMPANY, POWER SUPPLY DESIGN (SHANGHAI) CO., LTD. |
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