CN104488709A - Method for culturing bulb tissues of tulbaghia violacea floral leaf - Google Patents
Method for culturing bulb tissues of tulbaghia violacea floral leaf Download PDFInfo
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- CN104488709A CN104488709A CN201410727844.6A CN201410727844A CN104488709A CN 104488709 A CN104488709 A CN 104488709A CN 201410727844 A CN201410727844 A CN 201410727844A CN 104488709 A CN104488709 A CN 104488709A
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- 240000009239 Tulbaghia violacea Species 0.000 title claims abstract description 48
- 235000000430 Tulbaghia violacea Nutrition 0.000 title claims abstract description 48
- 238000000034 method Methods 0.000 title claims abstract description 30
- 238000012258 culturing Methods 0.000 title abstract description 4
- 230000006698 induction Effects 0.000 claims abstract description 24
- 238000005286 illumination Methods 0.000 claims abstract description 17
- 230000001939 inductive effect Effects 0.000 claims abstract description 11
- 230000001954 sterilising effect Effects 0.000 claims abstract description 7
- 239000000843 powder Substances 0.000 claims abstract description 5
- 229920001817 Agar Polymers 0.000 claims abstract description 4
- 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 abstract description 4
- 229930006000 Sucrose Natural products 0.000 claims abstract description 4
- 239000008272 agar Substances 0.000 claims abstract description 4
- 239000002609 medium Substances 0.000 claims description 19
- 230000035755 proliferation Effects 0.000 claims description 15
- 241000196324 Embryophyta Species 0.000 claims description 13
- 210000001519 tissue Anatomy 0.000 claims description 10
- 238000000338 in vitro Methods 0.000 claims description 9
- 239000012869 germination medium Substances 0.000 claims description 6
- 238000011081 inoculation Methods 0.000 claims description 6
- 238000004659 sterilization and disinfection Methods 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 238000005520 cutting process Methods 0.000 claims description 5
- 230000003020 moisturizing effect Effects 0.000 claims description 5
- 239000000701 coagulant Substances 0.000 claims description 3
- 210000002615 epidermis Anatomy 0.000 claims description 3
- 239000011159 matrix material Substances 0.000 claims description 3
- 239000006870 ms-medium Substances 0.000 claims description 3
- 238000012856 packing Methods 0.000 claims description 3
- 239000005720 sucrose Substances 0.000 claims description 3
- 238000012545 processing Methods 0.000 claims description 2
- 238000004161 plant tissue culture Methods 0.000 abstract description 5
- 230000001902 propagating effect Effects 0.000 abstract description 2
- 230000004083 survival effect Effects 0.000 abstract description 2
- 239000001963 growth medium Substances 0.000 abstract 7
- 230000035784 germination Effects 0.000 abstract 1
- 238000011031 large-scale manufacturing process Methods 0.000 abstract 1
- 239000012883 rooting culture medium Substances 0.000 abstract 1
- 229960004793 sucrose Drugs 0.000 abstract 1
- 206010020649 Hyperkeratosis Diseases 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 230000008929 regeneration Effects 0.000 description 3
- 238000011069 regeneration method Methods 0.000 description 3
- 230000007226 seed germination Effects 0.000 description 3
- 239000007858 starting material Substances 0.000 description 3
- 238000013459 approach Methods 0.000 description 2
- RCTYPNKXASFOBE-UHFFFAOYSA-M chloromercury Chemical compound [Hg]Cl RCTYPNKXASFOBE-UHFFFAOYSA-M 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 239000003599 detergent Substances 0.000 description 2
- 230000000763 evoking effect Effects 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 230000008774 maternal effect Effects 0.000 description 2
- 230000000384 rearing effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000011435 rock Substances 0.000 description 2
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- 240000006108 Allium ampeloprasum Species 0.000 description 1
- 235000005254 Allium ampeloprasum Nutrition 0.000 description 1
- 235000018714 Allium canadense Nutrition 0.000 description 1
- 235000005336 Allium ursinum Nutrition 0.000 description 1
- 244000109568 Allium vineale Species 0.000 description 1
- 235000005534 Allium vineale ssp. compactum Nutrition 0.000 description 1
- 235000003686 Allium vineale ssp. vineale Nutrition 0.000 description 1
- 241000234270 Amaryllidaceae Species 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 241000006479 Cyme Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241001634093 Lilium nanum Species 0.000 description 1
- 244000223014 Syzygium aromaticum Species 0.000 description 1
- 235000016639 Syzygium aromaticum Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000012090 tissue culture technique Methods 0.000 description 1
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 description 1
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention relates to a method for culturing bulb tissues of a tulbaghia violacea floral leaf and belongs to the field of plant tissue culture. The method comprises the following steps: preparing a culture medium; selecting and sterilizing an explant; germinating a bulb; inducing and propagating an adventitious bud; inducing rooting; and acclimating and transplanting a tissue culture seedling. A basic culture medium is an MS culture medium which contains 25g/L to 35g/L of cane sugar and 8g/L to 9g/L of agar powder; the pH of the culture medium is 5.6 to 5.8; a bulb germination culture medium contains 0.1mg/L to 0.5mg/L of MS+6-BA and 0.1mg/L to 0.2mg/L of NAA; an adventitious bud induction culture medium contains 3.0mg/L to 6.0mg/L of MS+6-BA and 0.05mg/L to 0.5mg/L of NAA; an adventitious bud propagation culture medium contains 0.5mg/L to 2.0mg/L of MS+6-BA and 0.05mg/L to 0.2mg/L of NAA; a rooting culture medium contains MS and 0.01mg/L to 0.5mg/L of IBA or 1/2 MS and 0.01mg/L to 0.5mg/L of IBA; the culture conditions in each stage are that the culture temperature is 25+/-2 DEG C; the illumination time is 2h/d to16h/d; the illumination intensity is 40mu.mol.m<-2>.s<-1> to 60mu.mol.m<-2>.s<-1>. The method disclosed by the invention has the advantages that the rapidly-propagated tulbaghia violacea floral leaf system seedling is high in propagation speed, strong, uniform, low in variation and high in transplanting survival rate, and the method is suitable for large-scale production of tulbaghia violacea floral leaf seedlings.
Description
Technical field
The present invention relates to plant tissue culture technique, be specifically related to a kind of method of floral leaf tulbaghia violacea bulb tissue cultures.
Background technology
Tulbaghia violacea (Tulbaghia violacea), the wild garlic that has another name called, African Lilium nanum Klotz. Et Garcke, belong to Amaryllidaceae herbaceos perennial, flowering bulb, plant height 30 ~ 50cm, the cylindrical clove of tool.Scape height about 40 ~ 60cm, the raw cyme in top opens purple powder little Hua, and the florescence is long, and actual cultivated species is often cultivated in flakes, and for excellent ground is by view flowers, complete stool all has leek taste, edible simultaneously.General tulbaghia violacea blade is entirely green, and the full edge of the present invention floral leaf used tulbaghia violacea blade has white lace, can enrich tulbaghia violacea germplasm resource storehouse, and flower is same with conventional tulbaghia violacea, but difficult solid, and whole plant growing way will be weaker than the entirely green tulbaghia violacea of blade.
The tradition breeding of floral leaf tulbaghia violacea is identical with traditional tulbaghia violacea, based on scale cuttage, plant division hyperplasia etc., but this method reproductive efficiency is low, according to Sun Lei etc. about floral leaf tulbaghia violacea patented technology (CN101869026A), utilize division propagation technology, floral leaf tulbaghia violacea offspring 2 is only propagation about 1 times, time less for germ plasm resource, is difficult to obtain a large amount of seedling at short notice.The tulbaghia violacea quick reproduction technique utilizing plant tissue culture technique to carry out the complete green kind of common blade has been reported, wish that will bravely waits (CN102144549B) to utilize bud for starting material, by callus of induce, then carrying out callus differentiation acquisition protocorm thus obtaining tulbaghia violacea regeneration plant; Chen Jianhua (CN101869068B) etc. also achieves tulbaghia violacea group culturation rapid propagating technology patent; In addition, He Yueqiu etc. (2010) research is also starting material with bud, is obtained the regeneration plant of tulbaghia violacea by callus of induce approach.And directly obtained the report of propagation adventitious bud technique by the fast numerous means of plant tissue culture about floral leaf tulbaghia violacea, have not yet to see.
Summary of the invention
The object of the invention is to the deficiency overcoming prior art existence, and a kind of method of floral leaf tulbaghia violacea bulb tissue cultures is provided.The sterilizable material that floral leaf tulbaghia violacea utilizes bulb induce to obtain, sets up good floral leaf tulbaghia violacea tissue culture regeneration system by direct evoking adventive bud approach, for floral leaf tulbaghia violacea excellent plantation expansion is numerous, rearing new variety etc. carries out technical support.
The object of the invention is to have come by following technical solution.The method of this floral leaf tulbaghia violacea bulb tissue cultures, the method comprises the steps:
1), the choosing and processing of explant: from floral leaf purple tender root, bulb is connected root and cut, cut off upper blade and bottom root, retain bulb length 2 ~ 3cm, peel off outer surface 1 layer of epidermis;
2), bulb is sprouted: on superclean bench, floral leaf tulbaghia violacea bulb through surface sterilization is being transferred in aseptic inoculation dish, the surface of the seed moisture is sucked with aseptic filter paper, after slightly cutting two end portions tissue, bulb is inoculated in germination medium MS+6-BA 0.1 ~ 0.5mg/L+NAA 0.1 ~ 0.2mg/L and cultivates, cultivation temperature 25 ± 2 DEG C, light application time is 12 ~ 16h/d, and intensity of illumination is 40 ~ 60 μm of olm
-2s
-1;
3) adventitious bud inducing: grow young leaves by what obtain, the aseptic seedling that 3 ~ 5cm is high is inoculated into adventitious bud induction culture base MS+6-BA 3.0 ~ 6.0mg/L+NAA 0.05 ~ 0.5mg/L after being divided into two in the middle of bulb on aseptic working platform and cutting open, carry out the induction of indefinite bud, cultivation temperature 25 ± 2 DEG C, light application time is 12 ~ 16h/d, and intensity of illumination is 40 ~ 60 μm of olm
-2s
-1;
4) adventitious bud proliferation: on aseptic working platform, induction is obtained cluster, indefinite bud that 2 ~ 3cm is high from base portion with 2 ~ 3 disjunctor indefinite buds for inoculation unit is separated, be inoculated in adventitious bud proliferation medium and breed, adventitious bud proliferation medium is MS+6-BA 0.5 ~ 2.0mg/L+NAA 0.05 ~ 0.2mg/L, cultivation temperature 25 ± 2 DEG C, light application time is 12 ~ 16h/d, and intensity of illumination is 40 ~ 60 μm of olm
-2s
-1;
5), adventitious bud rooting induction: on aseptic working platform, Seedling height is cut into single to the indefinite bud of growing thickly of 3 ~ 4cm from base portion, the induction of adventive root is carried out in access root media, root media is MS+IBA 0.01 ~ 0.5mg/L or 1/2MS+IBA 0.01 ~ 0.5mg/L, cultivation temperature 25 ± 2 DEG C, light application time is 12 ~ 16h/d, and intensity of illumination is 40 ~ 60 μm of olm
-2s
-1;
6), plantlet in vitro domestication and transplanting: after indefinite bud base portion grows the adventive root of 2 ~ 32 ~ 3cm, seedling of taking root moves to greenhouse, scattered light opens bottle cap after cultivating 5d, cultivate 1 ~ 2d again, the plant that taken root by floral leaf tulbaghia violacea is carefully taken out from blake bottle, cleans the medium on plantlet in vitro surface with running water, by floral leaf tulbaghia violacea plantlet in vitro, plant in matrix, 15d is cultivated in moisturizing more than 90%, then normally cultivates in greenhouse;
Minimal medium selects MS medium, and dosage of sucrose is 25 ~ 30g/L, and coagulating agent is agar powder, and consumption is 8 ~ 9g/L, is adjusted to 5.6 ~ 5.8 before medium pH packing; Bulb germination medium is MS+6-BA 0.1 ~ 0.5mg/L+NAA 0.1 ~ 0.2mg/L; Adventitious bud induction culture base is MS+6-BA 3.0 ~ 6.0mg/L+NAA 0.05 ~ 0.5mg/L; Adventitious bud proliferation medium is MS+6-BA 0.5 ~ 2.0mg/L+NAA 0.05 ~ 0.2mg/L; Root media is MS+IBA 0.01 ~ 0.5mg/L or 1/2MS+IBA 0.01 ~ 0.5mg/L; Bulb is sprouted, adventitious bud inducing and propagation, adventitious bud rooting condition of culture are that cultivation temperature 25 ± 2 DEG C, light application time is 12 ~ 16h/d, and intensity of illumination is 40 ~ 60 μm of olm
-2s
-1.
Before the selection of initial explant and process, bulb sprouting, adventitious bud inducing, adventitious bud proliferation, root induction, transplanting domestication process and after transplanting moisturizing more than 90% cultivate 15d.
Beneficial effect of the present invention is:
1, floral leaf tulbaghia violacea quick reproduction technique is carried out by tissue culture technique, produce by the restriction of area, season, weather and maternal plant growth year, be convenient to factorial seedling growth, can produce according to order, production time and scale controlled, for applying of floral leaf tulbaghia violacea provides sufficient high quality seedling guarantee.
2, reproduction coefficient reaches 2.5 ~ 3.0, rooting rate 100%, transplanting survival rate 100%, reaches the requirement that seedling factorial seedling growth is produced.
3, the present invention adopts the Reproduction methods of the direct evoking adventive bud of floral leaf tulbaghia violacea bulb, and without more organizing wound induction and callus induction link, greatly can reduce the generation of progeny variation in plant tissue culture process, seedling can keep maternal merit to greatest extent.
4, be conducive to floral leaf tulbaghia violacea fine individual plant, new varieties tissue-culturing rapid propagation Establishing is used for reference, promote.
5, set up good tissue-culturing rapid propagation system, for utilizing Plant Biotechnology from now on, the research work such as rearing new variety, genetic improvement is carried out to floral leaf tulbaghia violacea and lay the foundation.
Embodiment
Below by embodiment, the present invention is further elaborated, and help is understood the present invention by embodiment better, but the present invention is not limited only to following embodiment.
The method of this floral leaf tulbaghia violacea bulb tissue cultures, the method comprises the steps:
1, medium and condition of culture: bulb sprouting, adventitious bud inducing, adventitious bud proliferation propagation, adventitious bud rooting induction minimal medium are MS medium, dosage of sucrose is 25 ~ 30g/L, coagulating agent is agar powder, and consumption is 8 ~ 9g/L, is adjusted to 5.6 ~ 5.8 before medium pH packing; Bulb germination medium is MS+6-BA 0.1 ~ 0.5mg/L+NAA 0.1 ~ 0.2mg/L; Adventitious bud induction culture base is MS+6-BA 3.0 ~ 6.0mg/L+NAA 0.05 ~ 0.5mg/L; Adventitious bud proliferation medium is MS+6-BA 0.5 ~ 2.0mg/L+NAA 0.05 ~ 0.2mg/L; Root media is MS+IBA 0.01 ~ 0.5mg/L or 1/2MS+IBA 0.01 ~ 0.5mg/L; Bulb is sprouted, adventitious bud inducing and propagation, adventitious bud rooting condition of culture are that cultivation temperature 25 ± 2 DEG C, light application time is 12 ~ 16h/d, and intensity of illumination is 40 ~ 60 μm of olm
-2s
-1.
2, the process of explant and sterilization: choose floral leaf tulbaghia violacea bulb as starting material, from floral leaf purple tender root, bulb is connected root to cut, cut off upper blade and bottom root, retain bulb length 2 ~ 3cm, peel off outer surface 1 layer of epidermis, the explant liquid detergent solution cleaning 30min handled well, period constantly stirs gently, then running water is through the liquid detergent cleaning solution of clean bulb remained on surface, on the superclean bench of sterilization, the floral leaf tulbaghia violacea bulb cleaned up is being transferred in aseptic conical flask, with 70 ~ 75% alcohol-pickled 60s, period constantly rocks conical flask gently, removal is attached to floral leaf tulbaghia violacea the surface of the seed bubble, pour out 75% alcohol, use 0.1%HgCl again
2solution soaks seed 10 ~ 15min, or is that 1%NaClO solution soaks seed 20 ~ 30min with effective chlorine density, constantly rocks conical flask gently, remove and be attached to floral leaf tulbaghia violacea bulb blibbing, pour out 0.1%HgCl between soak period
2solution or 1%NaClO solution, with the bulb 4 ~ 5 time of sterile water wash through surface sterilization process, a small amount of sterile water of the seed after process soaks, for subsequent use.
3, seed germination: on superclean bench, floral leaf tulbaghia violacea seed through surface sterilization is transferred in aseptic inoculation dish, the surface of the seed moisture is blotted with aseptic filter paper, be inoculated in seed germination medium MS+6-BA 0.1 ~ 0.5mg/L+NAA 0.1 ~ 0.2mg/L and carry out seed germination, cultivation temperature 25 ± 2 DEG C, light application time is 12 ~ 16h/d, and intensity of illumination is 40 ~ 60 μm of olm
-2s
-1.
4, adventitious bud inducing: grow young leaves by what obtain, the aseptic seedling that 3 ~ 5cm is high is inoculated into adventitious bud induction culture base MS+6-BA 3.0 ~ 6.0mg/L+NAA 0.05 ~ 0.5mg/L after being divided into two in the middle of bulb on aseptic working platform and cutting open, carry out the induction of indefinite bud, cultivation temperature 25 ± 2 DEG C, light application time is 12 ~ 16h/d, and intensity of illumination is 40 ~ 60 μm of olm
-2s
-1.
5, adventitious bud proliferation: on aseptic working platform, induction is obtained cluster, indefinite bud that 2 ~ 3cm is high from base portion with 2 ~ 3 disjunctor indefinite buds for inoculation unit is separated, be inoculated in adventitious bud proliferation medium and breed, adventitious bud proliferation medium is MS+6-BA 0.5 ~ 2.0mg/L+NAA 0.05 ~ 0.2mg/L, cultivation temperature 25 ± 2 DEG C, light application time is 12 ~ 16h/d, and intensity of illumination is 40 ~ 60 μm of olm
-2s
-1.
6, adventitious bud rooting induction: on aseptic working platform, Seedling height is cut into single to the indefinite bud of growing thickly of 3 ~ 4cm from base portion, the induction of adventive root is carried out in access root media, root media is MS+IBA 0.01 ~ 0.5mg/L or 1/2MS+IBA 0.01 ~ 0.5mg/L, cultivation temperature 25 ± 2 DEG C, light application time is 12 ~ 16h/d, and intensity of illumination is 40 ~ 60 μm of olm
-2s
-1.
7, plantlet in vitro domestication and transplanting: after indefinite bud base portion grows the adventive root of 2 ~ 32 ~ 3cm, seedling of taking root moves to greenhouse, scattered light opens bottle cap after cultivating 5d, cultivate 1 ~ 2d again, the plant that taken root by floral leaf tulbaghia violacea is carefully taken out from blake bottle, cleans the medium on plantlet in vitro surface with running water, by floral leaf tulbaghia violacea plantlet in vitro, plant in matrix, 15d is cultivated in moisturizing more than 90%, then normally cultivates in greenhouse.
Claims (2)
1. a method for floral leaf tulbaghia violacea bulb tissue cultures, is characterized in that: the method comprises the steps:
1), the choosing and processing of explant: from floral leaf purple tender root, bulb is connected root and cut, cut off upper blade and bottom root, retain bulb length 2 ~ 3cm, peel off outer surface 1 layer of epidermis;
2), bulb is sprouted: on superclean bench, floral leaf tulbaghia violacea bulb through surface sterilization is being transferred in aseptic inoculation dish, the surface of the seed moisture is sucked with aseptic filter paper, after slightly cutting two end portions tissue, bulb is inoculated in germination medium MS+6-BA 0.1 ~ 0.5mg/L+NAA 0.1 ~ 0.2mg/L and cultivates, cultivation temperature 25 ± 2 DEG C, light application time is 12 ~ 16h/d, and intensity of illumination is 40 ~ 60 μm of olm
-2s
-1;
3) adventitious bud inducing: grow young leaves by what obtain, the aseptic seedling that 3 ~ 5cm is high is inoculated into adventitious bud induction culture base MS+6-BA 3.0 ~ 6.0mg/L+NAA 0.05 ~ 0.5mg/L after being divided into two in the middle of bulb on aseptic working platform and cutting open, carry out the induction of indefinite bud, cultivation temperature 25 ± 2 DEG C, light application time is 12 ~ 16h/d, and intensity of illumination is 40 ~ 60 μm of olm
-2s
-1;
4) adventitious bud proliferation: on aseptic working platform, induction is obtained cluster, indefinite bud that 2 ~ 3cm is high from base portion with 2 ~ 3 disjunctor indefinite buds for inoculation unit is separated, be inoculated in adventitious bud proliferation medium and breed, adventitious bud proliferation medium is MS+6-BA 0.5 ~ 2.0mg/L+NAA 0.05 ~ 0.2mg/L, cultivation temperature 25 ± 2 DEG C, light application time is 12 ~ 16h/d, and intensity of illumination is 40 ~ 60 μm of olm
-2s
-1;
5), adventitious bud rooting induction: on aseptic working platform, Seedling height is cut into single to the indefinite bud of growing thickly of 3 ~ 4cm from base portion, the induction of adventive root is carried out in access root media, root media is MS+IBA 0.01 ~ 0.5mg/L or 1/2MS+IBA 0.01 ~ 0.5mg/L, cultivation temperature 25 ± 2 DEG C, light application time is 12 ~ 16h/d, and intensity of illumination is 40 ~ 60 μm of olm
-2s
-1;
6), plantlet in vitro domestication and transplanting: after indefinite bud base portion grows the adventive root of 2 ~ 32 ~ 3cm, seedling of taking root moves to greenhouse, scattered light opens bottle cap after cultivating 5d, cultivate 1 ~ 2d again, the plant that taken root by floral leaf tulbaghia violacea is carefully taken out from blake bottle, cleans the medium on plantlet in vitro surface with running water, by floral leaf tulbaghia violacea plantlet in vitro, plant in matrix, 15d is cultivated in moisturizing more than 90%, then normally cultivates in greenhouse;
Minimal medium selects MS medium, and dosage of sucrose is 25 ~ 30g/L, and coagulating agent is agar powder, and consumption is 8 ~ 9g/L, is adjusted to 5.6 ~ 5.8 before medium pH packing; Bulb germination medium is MS+6-BA 0.1 ~ 0.5mg/L+NAA 0.1 ~ 0.2mg/L; Adventitious bud induction culture base is MS+6-BA 3.0 ~ 6.0mg/L+NAA 0.05 ~ 0.5mg/L; Adventitious bud proliferation medium is MS+6-BA 0.5 ~ 2.0mg/L+NAA 0.05 ~ 0.2mg/L; Root media is MS+IBA 0.01 ~ 0.5mg/L or 1/2MS+IBA 0.01 ~ 0.5mg/L; Bulb is sprouted, adventitious bud inducing and propagation, adventitious bud rooting condition of culture are that cultivation temperature 25 ± 2 DEG C, light application time is 12 ~ 16h/d, and intensity of illumination is 40 ~ 60 μm of olm
-2s
-1.
2. the method for floral leaf tulbaghia violacea bulb tissue cultures according to claim 1, is characterized in that: before the selection of initial explant and process, bulb sprouting, adventitious bud inducing, adventitious bud proliferation, root induction, transplanting domestication process and after transplanting moisturizing more than 90% cultivate 15d.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111264390A (en) * | 2020-03-16 | 2020-06-12 | 北华大学 | Method for regenerating plant by somatic cells of allium victorialis |
| CN112616666A (en) * | 2020-12-29 | 2021-04-09 | 浙江省农业科学院 | Tissue culture and rapid propagation method of Convolvulus verticillata |
| CN115474552A (en) * | 2022-11-02 | 2022-12-16 | 浙江省农业科学院 | Tissue culture method for yellow She Monian hemp plant |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101869068B (en) * | 2009-04-24 | 2012-01-04 | 上海上房园林植物研究所 | Tissue culture method for tulbaghia violacea |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101869068B (en) * | 2009-04-24 | 2012-01-04 | 上海上房园林植物研究所 | Tissue culture method for tulbaghia violacea |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111264390A (en) * | 2020-03-16 | 2020-06-12 | 北华大学 | Method for regenerating plant by somatic cells of allium victorialis |
| CN112616666A (en) * | 2020-12-29 | 2021-04-09 | 浙江省农业科学院 | Tissue culture and rapid propagation method of Convolvulus verticillata |
| CN112616666B (en) * | 2020-12-29 | 2021-12-07 | 浙江省农业科学院 | Tissue culture and rapid propagation method of Convolvulus verticillata |
| CN115474552A (en) * | 2022-11-02 | 2022-12-16 | 浙江省农业科学院 | Tissue culture method for yellow She Monian hemp plant |
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| CN104488709B (en) | 2016-04-20 |
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