CN102668985B - Rapid reproduction method for angelica keiskei koidzumi - Google Patents
Rapid reproduction method for angelica keiskei koidzumi Download PDFInfo
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- CN102668985B CN102668985B CN 201210168597 CN201210168597A CN102668985B CN 102668985 B CN102668985 B CN 102668985B CN 201210168597 CN201210168597 CN 201210168597 CN 201210168597 A CN201210168597 A CN 201210168597A CN 102668985 B CN102668985 B CN 102668985B
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- angelica keiskei
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Abstract
The invention discloses a rapid reproduction method for angelica keiskei koidzumi. The rapid reproduction method includes taking leaf blades of the angelica keiskei koidzumi as an explant, directly inducting and culturing the explant in vitro without a callus stage to produce cluster buds, and using the cluster buds to induct roots to grow into complete plant. According to the rapid reproduction method for the angelica keiskei koidzumi, the callus process is avoided, induction time is saved, induced mutation rate is reduced, raw material basis of development of medical value of the angelica keiskei koidzumi is provided, and application prospects are good.
Description
Technical field
The present invention relates to a kind of Angelica keiskei method for quickly breeding, being specifically related to a kind of blade of take Angelica keiskei is explant, induces the method for direct generation Multiple Buds on medium, belongs to field of plant tissue culture technique.
Background technology
Angelica keiskei (Angelica Keiskei Miq.Koidz), another name tomorrow leaf, eight zhang grass, originate in Japanese Ba Zhang island and Izu Islands.The frequent locals of edible Angelica keiskei, long-lived person is a lot, and to advanced age still very healthy, can work, so be called again long-life vegetable the samely with the young man.In recent years, the many compositions in Angelica keiskei are little by little understood, and are also therefore attracted attention, and wherein contain very abundant various vitamins, mineral matter and trace element, and these nutrient components are very balanced.The precious active component chalcone contained in Angelica keiskei has antitumor, anti-oxidant and removes the effects such as free radical, anti-gastric-ulcer, antibiotic and anti-inflammatory, so high edibility and the medical value of Angelica keiskei are subject to people's attention day by day.
Angelica keiskei is perennial herb, plant height 50-100cm; The leaf alternate, trifoliolate leaf, leaf margin tool serration; Floral white, give birth on the umbel top, and germination rate is low, and this also just makes tissue culture technique play great role in the breeding of Angelica keiskei.
Chinese patent (CN1930952) has been reported a kind of tissue cultivation and method for quickly breeding of Angelica keiskei, the method comprises: (1) adopts the stem with bud of Angelica keiskei as explant, carry out inoculated and cultured after sterilization, obtain callus, induce the shape bud of growing thickly; (2) propagation in medium by Multiple Buds, breed approximately 3 times by it; (3) will breed the Multiple Buds cutting of gained, and move and receive short its in root media and take root, obtain complete regeneration plant; (4) by Transplantation of Regenerated Plantlets in Nutrition Soil, obtain the transplanted seedling of Angelica keiskei.
The inventor finds by great many of experiments, and the method for inducing the blade of Angelica keiskei directly to produce Multiple Buds has reduced the evoked callus link, saves induction time, reduces the induced mutation rate, has good application prospect.
Summary of the invention
The purpose of this invention is to provide a kind of simple, quick method of inducing Angelica keiskei high frequency generation Multiple Buds that also can reduce the induced mutation rate, to overcome the main deficiency of prior art.
Implementation procedure of the present invention is as follows:
A kind of Angelica keiskei method for quickly breeding, its blade that utilizes Angelica keiskei is as explant, in vitro, without the callus stage, directly induces to cultivate to produce Multiple Buds, recycles afterwards inducing clumping bud and takes root and grow up to whole plant
.
Above-mentioned Angelica keiskei method for quickly breeding specifically comprises the following steps:
(1) draw materials: get fresh blade on the Angelica keiskei strain;
(2) sterilizing: blade HgCl
2soak sterilizing;
(3) inoculate and induce: the blade of the Angelica keiskei after sterilizing being cut into to the square fragment of 0.5～1.0cm, being seeded on the MS medium that induced bundle sprouts, cultivate 20d and produce green bud point, continue to cultivate 10～15d differentiation and grow up to Multiple Buds;
(4) take root: treat that Multiple Buds grows to 2～3cm, cut, insert in the MS root media that contains NAA 0.5mg/L and carry out culture of rootage.
In above-mentioned steps (2), with flowing water, rinse blade 3～5min, then use alcohol-pickled 30～60s of 70%, then use 0.1% HgCl
2soak sterilizing 5～7 min, do not stop to rock the bubble reduced on explant, make blade be immersed in sufficiently sterilised in mercuric chloride fully, finally use aseptic water washing 6～8 times.
In above-mentioned steps (3), the MS medium that described induced bundle is sprouted consists of: add 1 mg/L 2 in the MS medium, 4-D and 0.2 mg/L 6-BA.
In above-mentioned steps (3) and (4) process, condition of culture is: intensity of illumination is 40 μ molm
-2s
-1, light application time 16h, temperature is 25 ℃ ± 2 ℃.
Advantage of the present invention and good effect: the present invention directly induces the blade of Angelica keiskei to produce Multiple Buds, avoid first producing this process of callus, saved induction time, reduced the induced mutation rate, for the exploitation of Angelica keiskei medical value provides the raw material basis, there is good application prospect.
The accompanying drawing explanation
Fig. 1 is the bud point picture (20d) of observing under stereomicroscope;
Fig. 2 is the Multiple Buds picture (30d) of observing under stereomicroscope;
Fig. 3 is for directly growing the picture of Multiple Buds at different leaves explant edge.
Embodiment
(1) draw materials: on well-grown Angelica keiskei strain, get comparatively fresh intact leaf as explant.
(2) sterilization: rinse blade 4 min with flowing water, then on superclean bench with 70% alcohol-pickled 25 s, then use 0.1% HgCl
2soak sterilizing 6 minutes, do not stop to rock, purpose is to reduce the bubble on explant, and blade is immersed in mercuric chloride fully, and sterilization fully, is finally used aseptic water washing 7 times.
(3) inoculate and induce: the blade of the Angelica keiskei that disinfects is cut into to the square fragment of 0.5～1.0 centimetre, be seeded in and contain 1 mg/L 2, on the MS inducing culture of 4-D and 0.2 mg/L6-BA, 20 d produce a large amount of green bud points (Fig. 1), continue to cultivate, bud point directly grows up to Multiple Buds (Fig. 2).Through statistics of data acquisition, the direct bud ratio of Angelica keiskei explant has reached (Fig. 3) more than 90%.The condition of culture of inducing clumping bud is: intensity of illumination 40 μ molm
-2s
-1, light application time 16h, temperature is 25 ℃ ± 2 ℃.
(5) take root: treat that indefinite bud grows to 2-3cm, will induce the indefinite bud obtained to cut, insert in the root induction medium of MS+ NAA 1mg/L and cultivated, temperature be that 25 ℃ ± 2 ℃, light intensity are 40 μ molm
-2s
-1, light application time 16h condition, rooting rate is 100%.Cultivate 2-3 and can obtain a large amount of groups of training seedlings after week.
Claims (1)
1. an Angelica keiskei method for quickly breeding is characterized in that comprising the following steps:
(1) draw materials: get fresh blade on the Angelica keiskei strain;
(2) sterilizing: rinse blade 3～5min with flowing water, then use alcohol-pickled 30～60s of 70%, then use 0.1% HgCl
2soak sterilizing 5～7 min, do not stop to rock the bubble reduced on explant, make blade be immersed in sufficiently sterilised in mercuric chloride fully, finally use aseptic water washing 6～8 times;
(3) inoculate and induce: the blade of the Angelica keiskei after sterilizing is cut into to the square fragment of 0.5～1.0cm, is seeded on the MS medium that induced bundle sprouts, cultivate 20d and produce green bud point, continue to cultivate 10～15d differentiation and grow up to Multiple Buds,
The MS medium that induced bundle is sprouted consists of: add 1 mg/L 2 in the MS medium, 4-D and 0.2 mg/L 6-BA; Condition of culture is: intensity of illumination is 40 μ molm
-2s
-1, light application time 16h, temperature is 25 ℃ ± 2 ℃;
(4) take root: treat that Multiple Buds grows to 2～3cm, cut, insert in the MS root media that contains NAA 0.5mg/L and carry out culture of rootage, condition of culture is: intensity of illumination is 40 μ molm
-2s
-1, light application time 16h, temperature is 25 ℃ ± 2 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201210168597 CN102668985B (en) | 2012-05-28 | 2012-05-28 | Rapid reproduction method for angelica keiskei koidzumi |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201210168597 CN102668985B (en) | 2012-05-28 | 2012-05-28 | Rapid reproduction method for angelica keiskei koidzumi |
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| CN102668985A CN102668985A (en) | 2012-09-19 |
| CN102668985B true CN102668985B (en) | 2013-12-25 |
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Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103404435A (en) * | 2013-08-06 | 2013-11-27 | 合肥市金山园林工程有限公司 | Tissue culture method of purslane |
| CN104126509B (en) * | 2014-07-28 | 2016-08-17 | 贵州师范大学 | Tomorrow base of leaf section tissue culture rapid propagating technology |
| CN104145689A (en) * | 2014-08-26 | 2014-11-19 | 河北农业大学 | Cuttage and rapid propagation method for angelica keiskei |
| 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 |
| CN114190273A (en) * | 2021-10-26 | 2022-03-18 | 中南民族大学 | Rapid propagation method of angelica keiskei |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2007037535A (en) * | 2005-07-01 | 2007-02-15 | Takara Bio Inc | Tissue culture method of angelica keiskei |
| CN1934927A (en) * | 2006-05-09 | 2007-03-28 | 但修伍 | Angelica keiskei koidzmi culture method |
| CN100431407C (en) * | 2006-09-28 | 2008-11-12 | 南京大学 | Crastinus leaf tissue culturing and fast propagation process |
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