CN104145817B - Angelica keiskei leaf tissue cultivates fast breeding technique - Google Patents
Angelica keiskei leaf tissue cultivates fast breeding technique Download PDFInfo
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- CN104145817B CN104145817B CN201410369167.5A CN201410369167A CN104145817B CN 104145817 B CN104145817 B CN 104145817B CN 201410369167 A CN201410369167 A CN 201410369167A CN 104145817 B CN104145817 B CN 104145817B
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Abstract
The invention discloses a kind of Angelica keiskei leaf tissue and cultivate fast breeding technique, choose Angelica keiskei blade, with running water, 75% alcoholic solution soaks, and aseptic water washing, puts into 0.1% mercuric chloride solution and sterilize, use aseptic water washing more afterwards; Stripping and slicing; Kind is cultivated 20-30 days paddle cutout directors and is gone out callus group in callus inducing medium; Again callus is rolled into a ball stripping and slicing, be forwarded on callus proliferation medium and cultivate 20-30 days to callus proliferation; By the callus stripping and slicing of propagation, transfer and in inducing clumping bud medium, to carry out cultivation differentiation in 30 days produce Multiple Buds; The callus of Multiple Buds base portion is excised, is divided into single bud, bud is inoculated in cultivation in root media and obtains regeneration plant seedling in 20 days; Domestication is practiced seedling and is transplanted into after one week in the coconut husk soil of sterilization treatment until seedling survives.The invention has the beneficial effects as follows that Angelica keiskei blade does propagating materials, cost is low, and material source is wide.
Description
Technical field
The invention belongs to technical field of plant cultivation, relate to Angelica keiskei leaf tissue and cultivate fast breeding technique.
Background technology
Angelica keiskei medicine-food two-purpose, the value especially in medicinal is known as extremely potential emerging vegetables by all circles especially.China introduced Angelica keiskei from Japan before more than 20 year, and domestic at present only have several company just to introduce a fine variety, and area under cultivation is very little, and raw material is very high in international market price, and demand is large.In process of production, namely the usual 1-2 after planting of Angelica keiskei plant enters breeding period, dead immediately after blossoming and bearing fruit.The Angelica keiskei florescence continues longer, and fruit ripening rate is lower, and when gathering, kernel maturing differs, and its fruit is diachenium, causes the percentage of seedgermination of Angelica keiskei extremely low, not easily cultivates on a large scale.Nowadays Angelica keiskei seedling breed and culture technique has become the bottleneck of restriction Angelica keiskei and processed goods development thereof, and in document about the seedling breeding of Angelica keiskei and cultivation report very few, cause holding at high price of Angelica keiskei and products thereof on market.
For expanding the industrial production scale of Angelica keiskei, there were some research reports in Angelica keiskei tissue cultures once.At present, the result of study delivered is divided into three kinds: one to be that the researchs such as Li Jia find that the petiole of Angelica keiskei and blade are all not suitable for carrying out tissue cultures as explant, just utilize stem with bud to carry out the cultivations such as the induction and differentiation of callus; Two is that the reports such as Lv Xiuli utilize Angelica keiskei seed to obtain the cultural method of aseptic seedling and then differentiation and proliferation; Three is that Guo Zhiyou etc. reports and utilizes petiole and blade to carry out the method for tissue cultures as explant.
This research is compared with the research of above-mentioned three, although be all the breeding of method utilizing the means of tissue cultures to build Angelica keiskei, but has great difference with three.First, compare with the research of Li Jia etc., blade has been broken in this research can not carry out the conclusion of inducing as explant, and the blade that stem with bud and this research institute select is compared, and material has restricted greatly; Secondly, study with Lv Xiuli etc. and compare, the material restricted gender of this research reduces greatly; Finally, compare with the result of study of Guo Zhiyou etc., although it is similar to draw materials, based on this result of study display MS, medium effect is better, and the report of Guo Zhiyou thinks N
6basal medium is applicable to carrying out culture effect obviously, and the basal medium that therefore both select has great difference.
Summary of the invention
The object of the present invention is to provide a kind of Angelica keiskei leaf tissue to cultivate fast breeding technique, solve Angelica keiskei percentage of seedgermination low, raw material deficiency, expensive problem on market.
The technical solution adopted in the present invention is carried out according to following steps:
(1) choose Angelica keiskei blade, with running water, put on aseptic operating platform for subsequent use;
(2) soak with 75% alcoholic solution, aseptic water washing 3 times, puts into 0.1% mercuric chloride solution sterilization, uses aseptic water washing afterwards again 5 times;
(3) blot the water of blade with aseptic filter paper, then cut;
(4) by blade inoculation in callus inducing medium, cultivate under being placed in 24 DEG C of incubator 2000lx light intensity, illumination condition is h light every day 16,8 h dark, cultivates 20-30 days paddle cutout directors and goes out callus group;
(5) again callus is rolled into a ball stripping and slicing, be forwarded on callus proliferation medium, cultivation temperature 25 DEG C, illumination condition is 16 h light, 8 h dark, intensity of illumination 3000Lx, cultivates 20-30 days to callus proliferation;
(6) will the callus stripping and slicing of propagation, transfer and to cultivate in inducing clumping bud medium, cultivation temperature 25 DEG C, illumination condition is 16 h light, 8 h dark, intensity of illumination 3000Lx, cultivates 30 days differentiation generation Multiple Buds;
(7) excised by the callus of Multiple Buds base portion, be divided into single bud, be inoculated in by bud in root media, cultivation temperature 25 DEG C, illumination condition is 16 h light, 8 h dark, intensity of illumination 3000Lx, cultivates and obtains regeneration plant seedling in 20 days;
(8) healthy and strong seedling is chosen in tissue culture bottle, move to natural environmental condition lower 3 days, open the domestication in 3 days of tissue culture bottle lid and practice seedling, be transplanted into after one week in the coconut husk soil of sterilization treatment through domestication hardening, on cover film, water, keep air humidity, film is opened gradually, until seedling survives after 1 week.
Further, described step 3, step 5, step 6 stripping and slicing are 1cm
2.
Further, described callus inducing medium is MS+1mg/L6-BA+3mg/LNAA; Described callus proliferation medium is MS+6-BA2mg/L+2,4-D1mg/L; Described inducing clumping bud medium is MS+6-BA1mg/L+NAA0.5mg/L; Described root media is 1/2MS+0.02mg/LNAA.
The invention has the beneficial effects as follows that Angelica keiskei blade does propagating materials, cost is low, and material source is wider.
Embodiment
Below in conjunction with embodiment, the present invention is described in detail.
The present invention adopts medium based on MS, configures the combination of different plant growth substance:
Callus inducing medium: MS+1mg/L6-BA+3mg/LNAA;
Callus proliferation medium: MS+6-BA2mg/L+2,4-D1mg/L;
Inducing clumping bud medium: MS+6-BA1mg/L+NAA0.5mg/L;
Root induction: 1/2MS+0.02mg/LNAA.
The present invention carries out according to following steps:
(1) take blade as explant, the preferred upper end of blade, young a little tender blade, but avoid gathering young leaves, after running water 15-20min, put on aseptic operating platform for subsequent use.
(2) at transfer room, take out after superclean bench soaks vegetable material 30s with 75% alcoholic solution, aseptic water washing 3 times, put into 0.1% mercuric chloride solution and to sterilize 7min, aseptic water washing 7 times.
(3) blot the water of blade with aseptic filter paper, be then cut into 1cm
2size.
(4) by blade inoculation in callus inducing medium, cultivate under being placed in 24 DEG C of incubator 2000lx light intensity, illumination condition is h light every day 16,8 h dark, cultivates within 20-30 days, to go out callus to paddle cutout director and roll into a ball and increase.
(5) again callus group is evenly cut into 1cm
2, be forwarded on callus proliferation medium, cultivation temperature about 25 DEG C, illumination condition is 16 h light, 8 h dark, intensity of illumination 3000Lx, cultivates 20-30 days callus and can breed to 6cm
2left and right.
(6) callus of propagation is cut into 1cm
2size, transfers and to cultivate in inducing clumping bud medium, cultivation temperature about 25 DEG C, illumination condition is 16 h light, 8 h dark, intensity of illumination 3000Lx, cultivate callus after 15-20 days and break up generation Multiple Buds again, Multiple Buds well-grown after 30 days.
(7) callus of Multiple Buds base portion is excised, be divided into single bud, bud is inoculated in root media, cultivation temperature about 25 DEG C, illumination condition is 16 h light, 8 h dark, intensity of illumination 3000Lx, cultivate the fractionated generation root of bastem after 20 days, well-grown, obtains regeneration plant seedling.
(8) healthy and strong seedling is chosen in tissue culture bottle, move to natural environmental condition lower 3 days, open the domestication in 3 days of tissue culture bottle lid and practice seedling, be transplanted in the coconut husk soil of sterilization treatment after one week through domestication hardening, above cover film, note watering, keep air humidity, open film gradually after 1 week, and reduce frequency of watering, survival rate can reach more than 94%.
The present invention cultivates the novel breeding of method building Angelica keiskei by leaf tissue, broken the production bottleneck caused because percentage of seedgermination is low, for large-scale production provides condition.
The above is only to better embodiment of the present invention, not any pro forma restriction is done to the present invention, every any simple modification done above embodiment according to technical spirit of the present invention, equivalent variations and modification, all belong in the scope of technical solution of the present invention.
Claims (2)
1. Angelica keiskei leaf tissue cultivates quick-breeding method, it is characterized in that carrying out according to following steps:
(1) choose Angelica keiskei blade, with running water, put on aseptic operating platform for subsequent use;
(2) soak with 75% alcoholic solution, aseptic water washing, puts into 0.1% mercuric chloride solution and sterilizes, use aseptic water washing more afterwards;
(3) water of blade is blotted with aseptic filter paper, then stripping and slicing;
(4) by blade inoculation in callus inducing medium, cultivate under being placed in 24 DEG C of incubator 2000Lx light intensity, illumination condition is h light every day 16,8 h dark, cultivates 20-30 days paddle cutout directors and goes out callus group;
(5) again callus is rolled into a ball stripping and slicing, be forwarded on callus proliferation medium, cultivation temperature 25 DEG C, illumination condition is 16 h light, 8 h dark, intensity of illumination 3000Lx, cultivates 20-30 days to callus proliferation;
(6) will the callus stripping and slicing of propagation, transfer and to cultivate in inducing clumping bud medium, cultivation temperature 25 DEG C, illumination condition is 16 h light, 8 h dark, intensity of illumination 3000Lx, cultivates 30 days differentiation generation Multiple Buds;
(7) excised by the callus of Multiple Buds base portion, be divided into single bud, be inoculated in by bud in root media, cultivation temperature 25 DEG C, illumination condition is 16 h light, 8 h dark, intensity of illumination 3000Lx, cultivates and obtains regeneration plant seedling in 20 days;
(8) choose healthy and strong seedling in tissue culture bottle, move to natural environmental condition lower 3 days, open tissue culture bottle lid 3 days domestication hardenings, be transplanted in the coconut husk soil of sterilization treatment after one week through domestication hardening, above cover film, water, keep air humidity, open film gradually after 1 week, until seedling survives;
Described callus inducing medium is MS+1mg/L6-BA+3mg/LNAA; Described callus proliferation medium is MS+6-BA2mg/L+2,4-D1mg/L; Described inducing clumping bud medium is MS+6-BA1mg/L+NAA0.5mg/L; Described root media is 1/2MS+0.02mg/LNAA.
2. cultivate quick-breeding method according to Angelica keiskei leaf tissue described in claim 1, it is characterized in that: described step (3), step (5), step (6) stripping and slicing are 1cm
2.
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| CN104145817B true CN104145817B (en) | 2016-03-16 |
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| CN104996027B (en) * | 2015-06-23 | 2017-03-22 | 上海市园林科学研究所 | Method for increasing seed-germination rate of angelica keiskei |
| CN107896555A (en) * | 2017-11-20 | 2018-04-13 | 焦杰彪 | A kind of method for promoting Angelica Keiskei Seed sprouting using agitating device |
| CN114190273A (en) * | 2021-10-26 | 2022-03-18 | 中南民族大学 | Rapid propagation method of angelica keiskei |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102657084A (en) * | 2012-05-07 | 2012-09-12 | 上海交通大学 | Method for quickly propagating angelica keiskei koidzmi |
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| JP2007037535A (en) * | 2005-07-01 | 2007-02-15 | Takara Bio Inc | Tissue culture method of angelica keiskei |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102657084A (en) * | 2012-05-07 | 2012-09-12 | 上海交通大学 | Method for quickly propagating angelica keiskei koidzmi |
Non-Patent Citations (2)
| Title |
|---|
| 明日叶叶的愈伤组织诱导和快速繁殖;郭治友等;《广东农业科学》;20101231(第7期);第70-72页 * |
| 明日叶的组织培养与快速繁殖;李佳等;《植物生理学通讯》;20061231;第42卷(第6期);第1142页 * |
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