CN101869069B - Tissue culturing method of luosifu crocosmia crocosmiflora - Google Patents
Tissue culturing method of luosifu crocosmia crocosmiflora Download PDFInfo
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- CN101869069B CN101869069B CN2009100499771A CN200910049977A CN101869069B CN 101869069 B CN101869069 B CN 101869069B CN 2009100499771 A CN2009100499771 A CN 2009100499771A CN 200910049977 A CN200910049977 A CN 200910049977A CN 101869069 B CN101869069 B CN 101869069B
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Abstract
The invention relates to a tissue cultivating method of luosifu crocosmia crocosmiflora, comprising the steps of acquisition of sterile materials, differentiation and proliferation of buds, cultivation of strong-seedling of adventitious bud, rooting cultivation, hardening seedling, transplanting and the like. Compared with the prior art, the invention greatly improves the reproduction speed of luosifu crocosmia crocosmiflora and the uniformity of seedlings, and can realize industrialized large-scale production of 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 luosifu crocosmia crocosmiflora.
Background technology
Luosifu crocosmia crocosmiflora is an Iridaceae Crocosmia crocosmiflora (Nichols.) N. E. Br-Tritonia crocosmaeflore Lemoine plant, originates in the south, Africa, is perennial herb, and bulb and stolon are arranged, and bulb is oblate, and brown fiber plasma membrane is arranged outward.The high about 50cm of terrestrial stem often has branch, and leaf is wire sword shape, and flower is funnel-form for orange red, and the multi-disc bouquet is arranged, and is fourth contact awl inflorescence, from pale yellowish green leafage, extracts out, and height is straggly, reasonable organization, and perianth tube is thin and slightly crooked, and sliver is carried out.The adolescence of Mars flower is midsummer, and it is without cease that the flowers are in blossom, is the good material of arranging flower border, flower bed.But as the external new varieties of introducing, luosifu crocosmia crocosmiflora is introduced a fine variety negligible amounts, and plant division is slow, and the seedling supply is restricted.
Summary of the invention
The object of the invention is exactly the method for tissue culture that a kind of luosifu crocosmia crocosmiflora of the regularity that improves reproduction speed and seedling is provided in order to overcome the defective that above-mentioned prior art exists.
The object of the invention can be realized through following technical scheme:
The method for tissue culture of luosifu crocosmia crocosmiflora is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
Take the other bottom set of the big bulb of luosifu crocosmia crocosmiflora in spring, remove the stem sheet of coated outside, wash 1-3h with running water; On superclean bench, utilize the alcohol immersion 10-50s of mass concentration for 70-75% successively again, volumetric concentration is that the mercury of 0.5-2 ‰ soaks 10-30min; Use aseptic water washing 4-6 time again; After utilizing aseptic filter paper to blot the moisture on bottom set surface, bottom set is cut into the sections of the band axillalry bud of 0.5-2cm, sections is inoculated on the bottom set inducing culture;
(2) differentiation of bud and propagation
Sections is inoculated in that 1-3 is after week on the bottom set inducing culture, and its base portion begins to expand, and the yellow green projection occurs; 3-4 can see the bud meristematic tissue on the sections after week, cultivates 1-2 month again, has the bottom set of 0.1-0.3cm size to grow; With carrying out enrichment culture in the bottom set cutting-out immigration bulb proliferated culture medium; The callus of bulb base portion is more, but bottom set still breeds comparatively fast, and it is good to grow;
(3) bottom set strong seedling culture
The bottom set that in the bottom set inducing culture, induces is clump shape and distributes, and every clump has 4-6 ball, bottom set is divided into Xiao Cong again; Every clump has 1-3 bud; To divide subcaespitose bottom set to go on the strong seedling culture base then, bottom set expands rapidly, can grow 0.3-0.5cm after 20-30 days;
(4) culture of rootage
Get the bottom set of 0.3-0.5cm, root induction in the root media is gone in switching, and the base section of bottom set seedling dissolves the root original hase of white after 8-15 days, can grow to 2-4cm after 25-35 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 90-100%;
(5) refining seedling and transplanting
At culture of rootage 30-35 days, when root system grows to 2-3cm, select the aseptic seedlings of well developed root system, robust growth; Refine seedling 1-3 days in indoor uncork, then seedling is taken out, clean the agar of its root; Plant in the seedbed in greenhouse domestication 25-40 days, can transplant outdoorly, and give rich water quality management.
Described bottom set inducing culture comprises MS+6-BA0.1-5.0mg/L+NAA0.1-0.5mg/L.
The preferred MS+6-BA5.0mg/L+NAA0.5mg/L of described bottom set inducing culture.
Described bulb proliferated culture medium comprises MS+6-BA0.5-5.0mg/L.
The preferred MS+6-BA3.0mg/L of described bulb proliferated culture medium.
Described strong seedling culture base comprises MS+6-BA1.0-3.0mg/L.
The preferred MS+6-BA2.0mg/L of described strong seedling culture base.
Described root media comprises MS+NAA0.05-0.2mg/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 luosifu crocosmia crocosmiflora and the regularity of seedling through tissue culture technique, can realize the batch production production in enormous quantities of growing seedlings.
Embodiment
Below in conjunction with specific embodiment the present invention is elaborated.
Buy and execute example 1
(1) acquisition of sterilizable material
Take the other bottom set of the big bulb of luosifu crocosmia crocosmiflora in spring, remove the stem sheet of coated outside after, with behind the running water flushing 1h on superclean bench; Utilizing mass concentration successively is 70% alcohol immersion 10s; Volumetric concentration is that 0.5 ‰ mercury soaks 10min, uses aseptic water washing again 4 times, utilize aseptic filter paper to blot the moisture on bottom set surface after; Bulb is cut into the sections of the band axillalry bud of 0.5cm, is inoculated on the bottom set inducing culture that comprises MS+6-BA0.1mg/L+NAA0.1mg/L;
(2) differentiation of bud and propagation
Be inoculated on the bottom set inducing culture, the base portion of sections begins to expand after 1 week, the yellow green projection occurs; The visible bud meristematic tissue in 3 week backs was cultivated 1 month again, had the bottom set of 0.1cm to grow; Bottom set downcut moved in the bulb proliferated culture medium that comprises MS+6-BA0.5mg/L carry out enrichment culture; The callus of bulb base portion is more, but bottom set still breeds comparatively fast, and it is good to grow;
(3) bulb strong seedling culture
In the bottom set that in the bottom set inducing culture, induces, every clump has 4 balls, bottom set is divided into Xiao Cong again; Every clump has 1 bud; To divide subcaespitose bottom set to go on the strong seedling culture base that comprises MS+6-BA1.0mg/L then, bottom set expands rapidly, can grow 0.3cm after 25 days;
(4) culture of rootage
Get the bottom set of 0.3cm, root induction in the root media of going into to comprise MS+NAA0.05mg/L of transferring, the seedling base section dissolves the root original hase of white after 8 days, can grow to 2cm after 25 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 90%;
(5) refining seedling and transplanting
Culture of rootage 30 days when root system grows to 2cm, is selected the aseptic seedling of well developed root system, robust growth, in indoor uncork refining seedling 1 day, then seedling is taken out, and cleans the agar of root, plants in the seedbed in greenhouse domestication 25 days, can transplant outdoorly, and gives rich water quality management.
The medium of above-mentioned various situation 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
Take the other bottom set of the big bulb of luosifu crocosmia crocosmiflora in spring, remove the stem sheet of coated outside after, with behind the running water flushing 2h on superclean bench; Utilizing mass concentration successively is 75% alcohol immersion 30s; Volumetric concentration is that 1 ‰ mercury soaks 15min, uses aseptic water washing again 5 times, utilize aseptic filter paper to blot the moisture on bottom set surface after; Bulb is cut into the sections of the band axillalry bud of 1cm, is inoculated on the bottom set inducing culture that comprises MS+6-BA5.0mg/L+NAA0.5mg/L;
(2) differentiation of bud and propagation
Be inoculated on the bottom set inducing culture, the base portion of sections begins to expand after 2 weeks, the yellow green projection occurs; The visible bud meristematic tissue in 3 week backs was cultivated 1 month again, had the bottom set of 0.3cm to grow; Bottom set downcut moved in the bulb proliferated culture medium that comprises MS+6-BA3mg/L carry out enrichment culture; The callus of bulb base portion is more, but bottom set still breeds comparatively fast, and it is good to grow;
(3) bulb strong seedling culture
In the bottom set that in the bottom set inducing culture, induces, every clump has 6 balls, bottom set is divided into Xiao Cong again; Every clump has 3 buds; To divide subcaespitose bottom set to go on the strong seedling culture base that comprises MS+6-BA2.0mg/L then, bottom set expands rapidly, can grow 0.5cm after 20 days;
(4) culture of rootage
Get the bottom set 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 10 days, can grow to 3cm after 30 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 100%;
(5) refining seedling and transplanting
Culture of rootage 30 days when root system grows to 3cm, is selected the aseptic seedling of well developed root system, robust growth, in indoor uncork refining seedling 3 days, then seedling is taken out, and cleans the agar of root, plants in the seedbed in greenhouse domestication 30 days, can transplant outdoorly, and gives rich water quality management.
The medium of above-mentioned various situation 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
Take the other bottom set of the big bulb of luosifu crocosmia crocosmiflora in spring, remove the stem sheet of coated outside after, with behind the running water flushing 3h on superclean bench; Utilizing mass concentration successively is 75% alcohol immersion 50s; Volumetric concentration is that 2 ‰ mercury soaks 30min, uses aseptic water washing again 6 times, utilize aseptic filter paper to blot the moisture on bottom set surface after; Bulb is cut into the sections of the band axillalry bud of 2cm, is inoculated on the bottom set inducing culture that comprises MS+6-BA5.0mg/L+NAA0.5mg/L;
(2) differentiation of bud and propagation
Be inoculated on the bottom set inducing culture, the base portion of sections begins to expand after 3 weeks, the yellow green projection occurs; The visible bud meristematic tissue in 4 week backs was cultivated 2 months again, had the bottom set of 0.2cm to grow; Bottom set downcut moved in the bulb proliferated culture medium that comprises MS+6-BA3mg/L carry out enrichment culture; The callus of bulb base portion is more, but bottom set still breeds comparatively fast, and it is good to grow;
(3) bulb strong seedling culture
In the bottom set that in the bottom set inducing culture, induces, every clump has 5 balls, bottom set is divided into Xiao Cong again; Every clump has 2 buds; To divide subcaespitose bottom set to go on the strong seedling culture base that comprises MS+6-BA3.0mg/L then, bottom set expands rapidly, can grow 0.4cm after 30 days;
(4) culture of rootage
Get the bottom set of 0.4cm, root induction in the root media of going into to comprise MS+NAA0.2mg/L of transferring, the seedling base section dissolves the root original hase of white after 15 days, can grow to 4cm after 35 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 96%;
(5) refining seedling and transplanting
Culture of rootage 35 days when root system grows to 3cm, is selected the aseptic seedling of well developed root system, robust growth, in indoor uncork refining seedling 3 days, then seedling is taken out, and cleans the agar of root, plants in the seedbed in greenhouse domestication 40 days, can transplant outdoorly, and gives rich water quality management.
The medium of above-mentioned various situation also comprises sucrose 40g/L, agar powder 8g/L, pH=6.0,26 ℃ of cultivation temperature, illumination 2500lx.
Claims (5)
1. the method for tissue culture of luosifu crocosmia crocosmiflora is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
Take the other bottom set of the big bulb of luosifu crocosmia crocosmiflora in spring, remove the stem sheet of coated outside, wash 1-3h with running water; On superclean bench, utilize the alcohol immersion 10-50s of mass concentration for 70-75% successively again, volumetric concentration is that the mercury of 0.5-2 ‰ soaks 10-30min; Use aseptic water washing 4-6 time again; After utilizing aseptic filter paper to blot the moisture on bottom set surface, bottom set is cut into the sections of the band axillalry bud of 0.5-2cm, sections is inoculated on the bottom set inducing culture;
(2) differentiation of bud and propagation
Sections is inoculated in that 1-3 is after week on the bottom set inducing culture, and its base portion begins to expand, and the yellow green projection occurs; 3-4 sees the bud meristematic tissue on the sections after week, cultivated 1-2 month again, has the bottom set of 0.1-0.3cm size to grow; With carrying out enrichment culture in the bottom set cutting-out immigration bulb proliferated culture medium; The callus of bulb base portion is more, but bottom set still breeds comparatively fast, and it is good to grow;
(3) bottom set strong seedling culture
The bottom set that in the bottom set inducing culture, induces is clump shape and distributes, and every clump has 4-6 ball, bottom set is divided into Xiao Cong again; Every clump has 1-3 bud; To divide subcaespitose bottom set to go on the strong seedling culture base then, bottom set expands rapidly, and is long to 0.3-0.5cm after 20-30 days;
(4) culture of rootage
Get the bottom set of 0.3-0.5cm, root induction in the root media is gone in switching, and the base section of bottom set seedling dissolves the root original hase of white after 8-15 days, grows to 2-4cm after 25-35 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 90-100%;
(5) refining seedling and transplanting
At culture of rootage 30-35 days, when root system grows to 2-3cm, select the aseptic seedlings of well developed root system, robust growth; Refine seedling 1-3 days in indoor uncork, then seedling is taken out, clean the agar of its root; Plant in the seedbed in greenhouse domestication 25-40 days, promptly transplant outdoorly, and give rich water quality management;
Described bottom set inducing culture comprises MS+6-BA0.1-5.0mg/L+NAA0.1-0.5mg/L;
Described bulb proliferated culture medium comprises MS+6-BA3.0mg/L;
Described strong seedling culture base comprises MS+6-BA1.0-3.0mg/L;
Described root media comprises MS+NAA0.05-0.2mg/L.
2. the method for tissue culture of luosifu crocosmia crocosmiflora according to claim 1 is characterized in that, described bottom set inducing culture comprises MS+6-BA5.0mg/L+NAA0.5mg/L.
3. the method for tissue culture of luosifu crocosmia crocosmiflora according to claim 1 is characterized in that, described strong seedling culture base comprises MS+6-BA2.0mg/L.
4. the method for tissue culture of luosifu crocosmia crocosmiflora according to claim 1 is characterized in that, described root media comprises MS+NAA0.1mg/L.
5. the method for tissue culture of luosifu crocosmia crocosmiflora according to claim 1 is characterized in that, each 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 respectively.
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| CN2009100499771A CN101869069B (en) | 2009-04-24 | 2009-04-24 | Tissue culturing method of luosifu crocosmia crocosmiflora |
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| CN2009100499771A CN101869069B (en) | 2009-04-24 | 2009-04-24 | Tissue culturing method of luosifu crocosmia crocosmiflora |
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| 刘正兰, 刘军.雄黄兰组织培养和快繁技术研究.《四川农业大学学报》.2008,第26卷(第1期),45-47. * |
| 樊璐,等.火星花的引种栽培与繁殖.《中国园艺学会球根花卉分会2008年会暨球根花卉产业发展研讨会论文集》.2008,52-54. * |
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Owner name: SHANGHAI SHANGFANG LANDSCAPE PLANT RESEARCH INSTIT Free format text: FORMER OWNER: SHANGHAI SHANGFANG LANDSCAPE PLANT INSTITUTE Effective date: 20140102 Owner name: STATE GRID SHANGHAI ELECTRIC POWER COMPANY POWER S Effective date: 20140102 |
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Effective date of registration: 20140102 Address after: 201114 Pujiang village, Pujiang Town, Shanghai, Minhang District Patentee after: SHANGHAI SHANGFANG GARDEN PLANT RESEARCH INSTITUTE CO., LTD. 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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