CN1918972A - Angelica tissue culture breeding method - Google Patents
Angelica tissue culture breeding method Download PDFInfo
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- CN1918972A CN1918972A CN 200510021504 CN200510021504A CN1918972A CN 1918972 A CN1918972 A CN 1918972A CN 200510021504 CN200510021504 CN 200510021504 CN 200510021504 A CN200510021504 A CN 200510021504A CN 1918972 A CN1918972 A CN 1918972A
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Abstract
The invention discloses an angelica tissue culturing and breeding method, which comprises the following steps: adopting angelica root as external growing bulk, sterilizing, grafting to induce callus, breeding, proceeding bud and root induction, forming entire plant sprout, planting the entire sprout in the substrate, adopting MS culture medium as base and 6-furfuryl adenine, 6-benzyl adenine, 2, 4-dichlorophenoxyacetic acid, fruitone, sucrose and active carbon as auxiliary compositions.
Description
Technical field:
The present invention relates to biological cultivation and breeding, particularly a kind of angelica tissue culture breeding method.
Background technology:
Notopterygium root is the perennial natural crude drugs of Umbelliferae (Umbelliferae), is the endemic plant species of China, is middle one of the medicinal material the most frequently used in Qiang's medicine of hiding.Notopterygium root is used as medicine with root-like stock and root, and flavor is hot, warm in nature, and effects such as diffusing exterior cold, wind-damp dispelling, sharp joint are arranged, cure mainly flu rheumatism, fever and headache, skin disturb itch, diseases such as geomantic omen edema, skin sore.China goes through an edition pharmacopeia notopterygium root is all recorded, according to one one of Pharmacopoeia of the People's Republic of China version in 2005, dry rhizome and root for Umbelliferae (Umbelliferae) herbaceos perennial notopterygium root Notopterygium incisum Ting ex H.T.Chang and notopterygium forbesii N.forbesiiBoissieu, developing history is long, and the medicinal material commodity have: silkworm Qiang, RHIZOMA SEU RADIX NOTOPTERYGII, a Qiang, bar Qiang and the king of Qiang.Sichuan is the genuine producing region of notopterygium root, and notopterygium root is produced based on N.incisum in the river, the pin whole nation and outlet, ground such as main product Ganzi, Aba, Liangshan Mountain San Zhou high altitude mountainous area more than 2700 meters.
For a long time; because its source relies on wild excavating fully; germ plasm resource is destroyed seriously, reserves sharply reduce, and along with domestic and international increase to the notopterygium root demand, imbalance between supply and demand becomes increasingly conspicuous especially in recent years; interests are ordered about in addition; a large amount of predatorinesses are excavated, and notopterygium root is become gradually be on the verge of plant, face the species forfeiture; the danger of resource exhaustion, carry out germplasm protection and enlarge notopterygium root breed imperative.
Plant tissue culture technique is that (cytothesis that certain individual organ or tissue has broken up becomes the genetic potential of complete individuality to the totipotency of utilizing cell.), get cell mass or tissue on the plant individual, by the medium of artificial preparation, make these cell masses or tissue form thousands of plant.It is significant aspect following three that rapid propagation in vitro is carried out in utilization group training: (1) reproduction speed is fast, is not subjected to the weather factor affecting, all can breed indoor throughout the year; (2) be convenient to keep good strains of seeds; (3) very important meaning is arranged aspect the good mutant or the good first generation of hybrid preserving and breed, but a strain has the notopterygium root mutant of clear superiority or offspring's large tracts of land after tissue culture of a tool clear superiority is promoted.
Other high mountain natural crude drugs, as existing bibliographical information such as the tissue culture of rhodiola root etc., but wild notopterygium root tissue culture is not appeared in the newspapers as yet.From the explant of field acquisition, its surface is microorganism such as breed bacteria and fungi easily, and is that have even long to organization internal (endophyte); In addition, the contained quinones of notopterygium root, aldehydes matter are more, and very easy oxidation browning is therefore all very crucial to the selection and the sterilization method of explant material.The present invention adopts method for tissue culture first, carries out the work of wild notopterygium root germplasm in-vitro propagate, and this carries out wild notopterygium root genetic improvement, promotes the factorial seedling growth of notopterygium root that a new way is provided for saving notopterygium root material in imminent danger.
Summary of the invention
The purpose of this invention is to provide and a kind ofly carry out notopterygium root quality saving and fast seedling-breeding method by tissue culture.
Technical scheme of the present invention is:
A kind of angelica tissue culture breeding method, it is characterized in that: the radical bud that sprouts with notopterygium root is an explant, after sterile-processed, inoculation is induced and is formed callus, breed, lure bud to cultivate and culture of rootage through callus again, grow up to complete plant seedling, complete seedling is transplanted in the seedling medium, grows up to normal notopterygium root plant.
Described angelica tissue culture breeding method comprises the following steps:
The selection of A, explant and disinfecting: the radical bud that sprouts with notopterygium root is an explant, with flowing water flushing 3～5 minutes, be 75% alcohol-pickled sterilization 45～60 seconds with concentration again, with aseptic deionized water flushing 2～3 times, it is 0.1%～0.2% mercuric chloride that explant after will washing again places concentration, soak sterilization 10～14 minutes, aseptic deionized water flushing 2～3 times, standby;
B, explant inoculation: will be cut into 0.1～0.3cm through explant standby in the steps A
3Fritter, be seeded on the callus inducing medium, incubation time is 30～35 days, expands the formation callus in the radical bud incision;
C, callus propagation: the callus that obtains among the step B is inoculated on the callus proliferated culture medium, and callus was bred 25～35 days, grew the callus of yellow, densification;
The bud of D, callus is induced: the callus that obtains among the step C is inoculated into lures on the bud medium, cultivated 35～45 days, the notopterygium root seedling occurs growing thickly;
E, culture of rootage: the notopterygium root seedling plant division of growing thickly with obtaining among the step D, change in the root media, to cultivate 20～30 days, the seedling base portion the short root of white occurs and grows complete root system;
F, plantlet of transplant: before the transplanting, to containing the seedling medium sterilization of vermiculite, peat, to seedling medium, need not hardening can directly survive, and grows up to normal notopterygium root plant with the complete little seedling direct transplantation that obtains in the step e.
Described explant is disinfected and is adopted 75% alcohol-pickled sterilization 45～60 seconds and 0.1%～0.2% mercuric chloride to soak dual twice sterilization method of 10～14 minutes of sterilization.
The callus of described densification is meant the callus that non-particulate shape is dispersed in.
The basic source plant of described notopterygium root is Umbelliferae herbaceos perennial notopterygium root Notopterygiumincisum Ting ex H.T.Chang and notopterygium forbesii N.forbesii Boissieu.
Described callus inducing medium is: MS, KT 0.1～2.5mg/L, 2, and 4-D 0.5～1.5mg/L, NAA 0.2～0.6mg/L, wherein: MS is conventional minimal medium Murashige﹠amp; Skoog1962, KT are 6-furfuryl group adenine (kinetin), 2, and 4-D is a 2,4 dichlorophenoxyacetic acid, NAA is a methyl.
Described callus proliferated culture medium is: MS, KT 0.5～2.5mg/L, 2, and 4-D 0.5～1.0mg/L, wherein: MS is conventional minimal medium Murashige﹠amp; Skoog 1962, and KT is a 6-furfuryl group adenine, and 2,4-D is a 2,4 dichlorophenoxyacetic acid.
The described bud medium that lures is: MS, 6-BA 1.5～4.5mg/L, NAA 0.2～0.5mg/L, and wherein: MS is conventional minimal medium Murashige﹠amp; Skoog 1962, and 6-BA is a 6-benzyladenine, and NAA is a methyl.
Described root media is: 1/2MS, NAA 0.01～0.05mg/L, 2～3% sucrose, 0.8% agar, and wherein: MS is conventional minimal medium Murashige﹠amp; Skoog 1962, and the macroelement concentration in this routine minimal medium is reduced by half obtains 1/2MS, and NAA is a methyl.
Described callus inducing medium, callus proliferated culture medium and luring all adds 0.7～1.0% agar, 3% sucrose and 0.1～0.5% active carbon in the bud medium.
The composition of described MS medium is routinely: ammonium nitrate (NH
4NO
3) 1650mg/L, potassium nitrate (KNO
3) 900mg/L, magnesium sulfate (MgSO
47H2O) 370mg/L, potassium dihydrogen phosphate (KH
2PO
4) 170mg/L, calcium chloride (CaCl
22H
2O) 440mg/L, manganese sulphate (MnSO
44H
2O) 22.3mg/L, zinc sulphate (ZnSO
47H
2O) 8.6mg/L, boric acid (H
3BO
3) 6.2mg/L, potassium iodide (KI) 0.83mg/L, sodium molybdate (Na
2MoO
42H
2O) 0.25mg/L, copper sulphate (CuSO
45H
2O) 0.025mg/L, cobalt chloride (CoCl
26H
2O) 0.025mg/L, ferrous sulfate (FeSO
47H
2O) 27.8mg/L, Na
2EDTA2H
2O 37.3mg/L, glycine 2mg/L, thiamine hydrochloride 0.1mg/L, puridoxine hydrochloride 0.5mg/L, nicotinic acid 0.5mg/L, inositol 100mg/L.
Described complete seedling is meant the differentiation of having finished bud and root, and has grown vanelets, can independently carry out the immature plant of autophyting growth.
The physicochemical property of described seedling medium is that total porosity is 75%, and the content of organic matter is 20%.
The invention has the advantages that: the present invention utilizes the tissue culture fast breeding of exsomatizing significant aspect following three: (1) reproduction speed is fast, is not subjected to the weather factor affecting, all can breed indoor throughout the year, promptly starts fast; (2) be convenient to keep good strains of seeds, promptly synchronism is good; (3) very important meaning is arranged aspect the good mutant or the good first generation of hybrid preserving and breed, but a strain has the notopterygium root mutant of clear superiority or offspring's large tracts of land after tissue culture of a tool clear superiority is promoted.
The used medium of the present invention brings out short, fast, the propagation frequency height of emerging of time, and it is neat especially to emerge, and need not hardening can directly survive, and easy operating carries out large-scale production.
The present invention adopts method for tissue culture first, carry out the work of wild notopterygium root germplasm in-vitro propagate, this is for saving notopterygium root germ plasm resource in imminent danger, carry out wild notopterygium root genetic improvement, promote the factorial seedling growth of notopterygium root that a new way is provided, the method for tissue culture of the high mutantion line of a kind of artificial propagation, germ plasm resource {in vitro} conservation, utilization and the seed selection pharmaceutical ingredient content that can carry out notopterygium root is provided.
Embodiment
Embodiment 1
A kind of angelica tissue culture breeding method comprises the following steps:
A, get the radical bud that notopterygium root Notopterygium incisum Ting ex H.T. Chang sprouts and make explant, with flowing water flushing 3～5 minutes, sterilized 50 seconds with 75% alcohol again, with aseptic deionized water flushing 2～3 times, add 0.1% mercuric chloride again, sterilized 14 minutes, aseptic deionized water flushing 2 times uses the aseptic filter paper suck dry moisture standby at last;
B, explant inoculation: on superclean bench, will be cut into 0.1～0.3cm through the explant of steps A
3Fritter, be seeded on the callus inducing medium, this medium is: MS+KT2.0mg/L+2,4-D 0.5mg/L+NAA 0.2mg/L, add 0.8% agar, 3% sucrose and 0.1% active carbon in the medium, after cultivating about 35 days, as seen expand the formation callus in the radical bud incision;
Described superclean bench, model are HS-1300, produce by SuZhou Antai Air Tech Co., Ltd. of Su Jing group,
Described MS is conventional minimal medium Murashige﹠amp; The abbreviation of Skoog 1962, KT are 6-furfuryl group adenine, 2, and 4-D is a 2,4 dichlorophenoxyacetic acid, NAA is a methyl;
C, callus propagation: the callus that obtains among the step B is inoculated on the callus proliferated culture medium, this medium is MS+KT 1.5mg/L+2,4-D 1.0mg/L, add 0.8% agar, 3% sucrose and 0.1% active carbon in the medium, behind the enrichment culture 30～32 days, grow the callus of yellow, densification;
The bud of D, callus is induced: the callus that obtains among the step C is inoculated into lures on the bud medium, this medium is MS+6-BA 4.0mg/L+NAA 0.2mg/L, add 0.8% agar, 3% sucrose and 0.1% active carbon in the medium, cultivate after 35～45 days, the notopterygium root seedling occurs growing thickly;
E, culture of rootage: when the notopterygium root seedling of growing thickly among the step D is grown to 2～3 centimetres, change root media over to, this medium is 1/2MS+NAA 0.01mg/L, adds 2% sucrose, 0.8% agar in the medium, cultivate after 20～30 days, the seedling base portion the short root of white occurs and grows complete root system;
Cultivation temperature and illumination condition: temperature is 21 ± 2 ℃, and intensity of illumination is 1600Lux, and light application time is 14 hours;
F, plantlet of transplant: before the transplanting, to containing the seedling medium sterilization of vermiculite, peat, the complete little seedling direct transplantation that obtains in the step e to seedling medium, be need not hardening and can directly survive and grow up to normal notopterygium root plant, generally survival rate is more than 80%.
The physicochemical property of described seedling medium is: total porosity is 75%, and the content of organic matter is 20%.
Embodiment 2
A kind of angelica tissue culture breeding method comprises the following steps:
A, get the radical bud that notopterygium root Notopterygium incisum Ting ex H.T.Chang sprouts and make explant, with flowing water flushing 3～5 minutes, sterilized 50 seconds with 75% alcohol again, with aseptic deionized water flushing 3 times, add 0.15% mercuric chloride again, sterilized 12 minutes, aseptic deionized water flushing 3 times uses the aseptic filter paper suck dry moisture standby at last;
B, explant inoculation: on superclean bench, will be cut into 0.1～0.3cm through the explant of steps A
3Fritter, be seeded on the callus inducing medium, this medium is: MS+KT1.0mg/L+2,4-D 0.8mg/L+NAA 0.4mg/L, add 1.0% agar, 3% sucrose and 0.5% active carbon in the medium, cultivate after 30～35 days, expand the formation callus in the radical bud incision;
Described superclean bench, model are HS-1300, are produced by SuZhou Antai Air Tech Co., Ltd. of Su Jing group;
Described MS is conventional minimal medium Murashige﹠amp; The abbreviation of Skoog 1962, KT are 6-furfuryl group adenine, 2, and 4-D is a 2,4 dichlorophenoxyacetic acid, NAA is a methyl;
C, callus propagation: the callus that obtains among the step B is inoculated on the callus proliferated culture medium, this medium is MS+KT 1.0mg/L+2,4-D 0.7mg/L, add 1.0% agar, 3% sucrose and 0.5% active carbon in the medium, behind the enrichment culture 35 days, grow the callus of yellow, densification;
The bud of D, callus is induced: the callus that obtains among the step C is inoculated into lures on the bud medium, this medium is MS+6-BA 3.0mg/L+NAA 0.3mg/l, add 1.0% agar, 3% sucrose and 0.5% active carbon in the medium, cultivate after 35～40 days,, the notopterygium root seedling occurs growing thickly;
E, culture of rootage: etc. the notopterygium root seedling of growing thickly among the step D when growing to 2～3 centimetres, change in the root media, this medium is 1/2MS+NAA 0.03mg/L, add 2% sucrose, 0.8% agar in the medium, cultivate after 20～30 days, the seedling base portion the short root of white occurs and grows complete root system;
Cultivation temperature and illumination condition: temperature is 21 ± 2 ℃, and intensity of illumination is 1600Lux, and light application time is 14 hours;
F, plantlet of transplant: the complete little seedling direct transplantation that obtains in the step e to the sterile-processed seedling medium that contains vermiculite, peat, is grown up to normal notopterygium root plant.
The physicochemical property of described seedling medium is: total porosity is 75%, and the content of organic matter is 20%.
Embodiment 3
A kind of angelica tissue culture breeding method comprises the following steps:
A, get the radical bud that notopterygium root Notopterygium incisum Ting ex H.T.Chang sprouts and make explant, with flowing water flushing 3～5 minutes, sterilized 45 seconds with 75% alcohol again, with aseptic deionized water flushing 2 times, add 0.2% mercuric chloride again, sterilized 12 minutes, aseptic deionized water flushing 3 times uses the aseptic filter paper suck dry moisture standby at last;
B, explant inoculation: on superclean bench, will be cut into 0.1～0.3cm through the explant of steps A
3Fritter, be seeded on the callus inducing medium, this medium is: MS+KT0.2mg/L+2,4-D 1.0mg/L+NAA 0.5mg/L, in medium, add 0.8% agar, 3% sucrose and 0.2% active carbon, cultivate after 30～35 days, expand the formation callus in the radical bud incision;
Described MS is conventional minimal medium Murashige﹠amp; The abbreviation of Skoog 1962, KT are 6-furfuryl group adenine, 2, and 4-D is a 2,4 dichlorophenoxyacetic acid, NAA is a methyl;
C, callus propagation: the callus that obtains among the step B is inoculated on the callus proliferated culture medium, this medium is MS+KT 2.0mg/L+2,4-D 1.0mg/L, add 0.8% agar, 3% sucrose and 0.2% active carbon in the medium, behind the enrichment culture 30～35 days, grow the callus of yellow, densification;
The bud of D, callus is induced: the callus that obtains among the step C is inoculated into lures on the bud medium, this medium is MS+6-BA 4.0mg/L+NAA 0.5mg/L, add 0.8% agar, 3% sucrose and 0.2% active carbon in the medium, cultivation through 35～40 days, the notopterygium root seedling occurs growing thickly;
E, culture of rootage: etc. the notopterygium root seedling of growing thickly among the step D when growing to 2～3 centimetres, change in the root media, this medium is 1/2MS+NAA 0.02mg/L, add 2% sucrose, 0.8% agar in the medium, cultivate after 20～30 days, the seedling base portion the short root of white occurs and grows complete root system;
Cultivation temperature and illumination condition: temperature is 21 ± 2 ℃, and intensity of illumination is 1600Lux, and light application time is 14 hours;
F, plantlet of transplant: before the transplanting, to containing the seedling medium sterilization of vermiculite, peat, the complete little seedling direct transplantation that obtains in the step e to seedling medium, is grown up to normal notopterygium root plant.
The physicochemical property of described seedling medium is: total porosity is 75%, and the content of organic matter is 20%.
Embodiment 4
A kind of angelica tissue culture breeding method comprises the following steps:
A, get the radical bud that notopterygium root Notopterygium forbesii Boissieu sprouts and make explant, with flowing water flushing 3～5 minutes, sterilized 55 seconds with 75% alcohol again, with aseptic deionized water flushing 3 times, add 0.15% mercuric chloride again, sterilized 12 minutes, aseptic deionized water flushing 3 times uses the aseptic filter paper suck dry moisture standby at last;
B, explant inoculation: on superclean bench, will be cut into 0.1-0.3cm through the explant of steps A
3Fritter, be seeded on the callus inducing medium, this medium is: MS+KT1.5mg/L+2,4-D 1.5mg/L+NAA 0.5mg/L, in medium, add 0.9% agar, 3% sucrose and 0.3% active carbon, cultivate after 30～35 days, expand the formation callus in the radical bud incision;
Described superclean bench, model are HS-1300, are produced by SuZhou Antai Air Tech Co., Ltd. of Su Jing group;
Described MS is conventional minimal medium Murashige﹠amp; The abbreviation of Skoog 1962, KT are 6-furfuryl group adenine, 2, and 4-D is a 2,4 dichlorophenoxyacetic acid, NAA is a methyl;
C, callus propagation: the callus that obtains among the step B is inoculated on the callus proliferated culture medium, this medium is MS+KT 2.5mg/L+2,4-D 0.9mg/L, in medium, add 0.9% agar, 3% sucrose and 0.3% active carbon, behind the enrichment culture 25～30 days, grow the callus of yellow, densification;
The bud of D, callus is induced: the callus that obtains among the step C is inoculated into lures on the bud medium, this medium is MS+6-BA 3.5mg/L+NAA 0.4mg/L, in medium, add 0.9% agar, 3% sucrose and 0.3% active carbon, cultivate after 35～45 days, the notopterygium root seedling occurs growing thickly;
E, culture of rootage: when the notopterygium root seedling of growing thickly among the step D is grown to 2～3 centimetres, change in the root media, this medium is 1/2MS+NAA 0.02mg/L, adds 2% sucrose, 0.8% agar in the medium, cultivated 20～30 days, the seedling base portion the short root of white occurs and grows complete root system;
Cultivation temperature and illumination condition: temperature is 21 ± 2 ℃; Intensity of illumination is 1600Lux; Light application time is 14 hours;
F, plantlet of transplant: the complete little seedling direct transplantation that obtains in the step e to the sterile-processed seedling medium that contains vermiculite, peat, is grown up to normal notopterygium root plant.
The physicochemical property of described seedling medium is: total porosity is 75%, and the content of organic matter is 20%.
Embodiment 5
A kind of angelica tissue culture breeding method comprises the following steps:
A, get the radical bud that notopterygium root Notopterygium forbesii Boissieu sprouts and make explant, with flowing water flushing 3～5 minutes, sterilized 45 seconds with 75% alcohol again, with aseptic deionized water flushing 2 times, add 0.2% mercuric chloride again, sterilized 12 minutes, aseptic deionized water flushing 3 times uses the aseptic filter paper suck dry moisture standby at last;
B, explant inoculation: on superclean bench, will be cut into 0.1～0.3cm through the explant of steps A
3Fritter, be seeded on the callus inducing medium, this medium is: MS+KT0.5mg/L+2,4-D 1.0mg/L+NAA 0.5mg/L, in medium, add 0.8% agar, 3% sucrose and 0.2% active carbon, cultivate after 30～35 days, expand the formation callus in the radical bud incision:
Described MS is conventional minimal medium Murashige﹠amp; The abbreviation of Skoog 1962, KT are 6-furfuryl group adenine, 2, and 4-D is a 2,4 dichlorophenoxyacetic acid, NAA is a methyl;
C, callus propagation: the callus that obtains among the step B is inoculated on the callus proliferated culture medium, this medium is MS+KT 2.0mg/L+2,4-D 1.0mg/L, in medium, add 0.8% agar, 3% sucrose and 0.2% active carbon, behind the enrichment culture 25～35 days, grow the callus of yellow, densification;
The bud of D, callus is induced: the callus that obtains among the step C is inoculated into lures on the bud medium, this medium is MS+6-BA 4.0mg/L+NAA 0.5mg/L, in medium, add 0.8% agar, 3% sucrose and 0.2% active carbon, cultivate after 35～40 days, the notopterygium root seedling occurs growing thickly;
E, culture of rootage: etc. the notopterygium root seedling of growing thickly among the step D when growing to 2～3 centimetres, change in the root media, this medium is 1/2MS+NAA 0.02mg/L, add 2% sucrose, 0.8% agar in the medium, cultivate after 20～30 days, the seedling base portion the short root of white occurs and grows complete root system;
Cultivation temperature and illumination condition: temperature is 21 ± 2 ℃; Intensity of illumination is 1600Lux; Light application time is 14 hours;
F, plantlet of transplant: the complete little seedling direct transplantation that obtains in the step e to the sterile-processed seedling medium that contains vermiculite, peat, is grown up to normal notopterygium root plant.
The physicochemical property of described seedling medium is: total porosity is 75%, and the content of organic matter is 20%.
Claims (9)
1, a kind of angelica tissue culture breeding method, it is characterized in that: the radical bud that sprouts with notopterygium root is an explant, after sterile-processed, inoculation is induced and is formed callus, breed, lure bud to cultivate and culture of rootage through callus again, form complete plant seedling, complete seedling is transplanted in the seedling medium, grows up to normal notopterygium root plant.
2, a kind of angelica tissue culture breeding method according to claim 1, it is characterized in that: described angelica tissue culture breeding method comprises the following steps:
The selection of A, explant and disinfecting: the radical bud that sprouts with notopterygium root is an explant, with flowing water flushing 3～5 minutes, be 75% alcohol-pickled sterilization 45～60 seconds with concentration again, with aseptic deionized water flushing 2～3 times, it is 0.1%～0.2% mercuric chloride that explant after will washing again places concentration, soak sterilization 10～14 minutes, aseptic deionized water flushing 2～3 times, standby;
B, explant inoculation: will be cut into 0.1～0.3cm through explant standby in the steps A
3Fritter, be seeded on the callus inducing medium, incubation time is 30～35 days, expands the formation callus in the radical bud incision;
C, callus propagation: the callus that obtains among the step B is inoculated on the callus proliferated culture medium, and callus was bred 25～35 days, grew the callus of yellow, densification;
The bud of D, callus is induced: the callus that obtains among the step C is inoculated into lures on the bud medium, cultivated 35～45 days, the notopterygium root seedling occurs growing thickly;
E, culture of rootage: the notopterygium root seedling plant division of growing thickly with obtaining among the step D, change in the root media, to cultivate 20～30 days, the seedling base portion the short root of white occurs and grows complete root system;
F, plantlet of transplant: before the transplanting, to containing the seedling medium sterilization of vermiculite, peat, to seedling medium, need not hardening can directly survive, and grows up to normal notopterygium root plant with the complete little seedling direct transplantation that obtains in the step e.
3, a kind of angelica tissue culture breeding method according to claim 1 and 2 is characterized in that: the basic source plant of described notopterygium root is Umbelliferae herbaceos perennial notopterygium root Notopterygiumincisum Ting ex H.T.Chang and notopterygium forbesii N.forbesii Boissieu.
4, a kind of angelica tissue culture breeding method according to claim 2, it is characterized in that: described callus inducing medium is MS, KT 0.1～2.5mg/L, 2,4-D 0.5～1.5mg/L, NAA 0.2～0.6mg/L, wherein: MS is conventional minimal medium Murashige﹠amp; Skoog 1962, and KT is a 6-furfuryl group adenine, and 2,4-D is a 2,4 dichlorophenoxyacetic acid, NAA is a methyl.
5, a kind of angelica tissue culture breeding method according to claim 2, it is characterized in that: described callus proliferated culture medium is: MS, KT 0.5～2.5mg/L, 2,4-D 0.5～1.0mg/L, wherein: MS is conventional minimal medium Murashige﹠amp; Skoog 1962, and KT is a 6-furfuryl group adenine, and 2,4-D is a 2,4 dichlorophenoxyacetic acid.
6, a kind of angelica tissue culture breeding method according to claim 2 is characterized in that: the described bud medium that lures is: MS, 6-BA 1.5～4.5mg/L, NAA 0.2～0.5mg/L, and wherein: MS is conventional minimal medium Murashige﹠amp; Skoog 1962, and 6-BA is a 6-benzyladenine, and NAA is a methyl.
7, a kind of angelica tissue culture breeding method according to claim 2, it is characterized in that: described root media is: 1/2MS, NAA 0.01～0.05mg/L, 2～3% sucrose, 0.8% agar, and wherein: MS is conventional minimal medium Murashige﹠amp; Skoog 1962, and the macroelement concentration in this routine minimal medium is reduced by half obtains 1/2MS, and NAA is a methyl.
8, a kind of angelica tissue culture breeding method according to claim 2 is characterized in that: described callus inducing medium, callus proliferated culture medium and lure the bud medium all to add 0.7～1.0% agar, 3% sucrose and 0.1～0.5% active carbon.
9, a kind of angelica tissue culture breeding method according to claim 1 and 2 is characterized in that: the physicochemical property of described seedling medium is that total porosity is 75%, and the content of organic matter is 20%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2005100215042A CN1918972B (en) | 2005-08-22 | 2005-08-22 | Angelica tissue culture breeding method |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102948322A (en) * | 2012-11-19 | 2013-03-06 | 四川新荷花中药饮片股份有限公司 | Seed propagation method for quickly obtaining healthy notopterygium incisum Ting ex H. T. Chang seedlings in abundance |
| CN107567967A (en) * | 2017-09-28 | 2018-01-12 | 惠州市支持网知识产权运营有限公司 | A kind of breeding method of notopterygium root |
| CN108719072A (en) * | 2018-06-21 | 2018-11-02 | 四川千草生物技术股份有限公司 | Rhizoma Et Radix Notopterygii seed tissue culture fast breeding technology |
| CN109652360A (en) * | 2019-01-11 | 2019-04-19 | 成都大学 | A kind of Rhizoma Et Radix Notopterygii cell culture processes obtaining high biological yield |
-
2005
- 2005-08-22 CN CN2005100215042A patent/CN1918972B/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102948322A (en) * | 2012-11-19 | 2013-03-06 | 四川新荷花中药饮片股份有限公司 | Seed propagation method for quickly obtaining healthy notopterygium incisum Ting ex H. T. Chang seedlings in abundance |
| CN102948322B (en) * | 2012-11-19 | 2014-04-16 | 四川新荷花中药饮片股份有限公司 | Seed propagation method for quickly obtaining healthy notopterygium incisum Ting ex H. T. Chang seedlings in abundance |
| CN107567967A (en) * | 2017-09-28 | 2018-01-12 | 惠州市支持网知识产权运营有限公司 | A kind of breeding method of notopterygium root |
| CN108719072A (en) * | 2018-06-21 | 2018-11-02 | 四川千草生物技术股份有限公司 | Rhizoma Et Radix Notopterygii seed tissue culture fast breeding technology |
| CN109652360A (en) * | 2019-01-11 | 2019-04-19 | 成都大学 | A kind of Rhizoma Et Radix Notopterygii cell culture processes obtaining high biological yield |
| CN109652360B (en) * | 2019-01-11 | 2022-04-15 | 成都大学 | Notopterygium incisum cell culture method for obtaining high biological yield |
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| CN1918972B (en) | 2010-10-27 |
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