CN102668985A

CN102668985A - Rapid reproduction method for angelica keiskei koidzumi

Info

Publication number: CN102668985A
Application number: CN2012101685971A
Authority: CN
Inventors: 黄萱; 芦恒; 刘梦昕; 曾洁; 黄凤智; 李颖
Original assignee: Northwest University
Current assignee: Northwest University
Priority date: 2012-05-28
Filing date: 2012-05-28
Publication date: 2012-09-19
Anticipated expiration: 2032-05-28
Also published as: CN102668985B

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

A kind of Angelica keiskei method for quickly breeding

Technical field

The present invention relates to a kind of Angelica keiskei method for quickly breeding, being specifically related to a kind of blade with Angelica keiskei is explant, on medium, induces the grow thickly method of bud of direct generation, 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 the eight zhang islands and the Izu Islands of Japan.The frequent locals of edible Angelica keiskei, long-lived person is a lot, and to advanced age still very healthy, can work with the young man, so be called long-life vegetable again the samely.In recent years, the many compositions in the Angelica keiskei are little by little understood, and are also therefore attracted attention, and wherein contain profuse various vitamin, mineral matter and trace element, and these nutrient components are very balanced.The precious active component chalcone that contains in the Angelica keiskei has antitumor, anti-oxidant and removes effects such as free radical, anti-gastric-ulcer, antibiotic and anti-inflammatory, so the high edibility of Angelica keiskei and medical value receive people's attention day by day.

Angelica keiskei is a 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 culture and method for quickly breeding of Angelica keiskei, and this method comprises: (1) adopts the stem with bud of Angelica keiskei as explant, after sterilization, carries out inoculated and cultured, obtains callus, induces the shape bud of growing thickly; (2), it is bred about 3 times through the propagation of bud in medium of growing thickly; (3) will breed the bud cutting of growing thickly of gained, and move and receive in the root media short its and take root, obtain complete regeneration plant; (4) regeneration plant is transplanted in the nutrition soil, obtains the transplanted seedling of Angelica keiskei.

The inventor finds through a large amount of experiments, induces the direct method that produces the bud of growing thickly of blade of Angelica keiskei to reduce the evoked callus link, practices thrift induction time, reduces the induced mutation rate, has good application prospects.

Summary of the invention

The purpose of this invention is to provide a kind of Angelica keiskei high frequency of inducing simple, quick and that can reduce the induced mutation rate and produce the method for the bud of growing thickly, to overcome the main deficiency of prior art.

Implementation procedure of the present invention is following:

A kind of Angelica keiskei method for quickly breeding, its blade that utilizes Angelica keiskei produce grow thickly bud without direct inducing culture of callus stage as explant under isolated condition, utilize inducing clumping bud to take root afterwards again and grow up to whole plant .

Above-mentioned Angelica keiskei method for quickly breeding specifically may further comprise the steps:

(1) draws materials: become to get fresh blade in the strain at Angelica keiskei;

(2) sterilization: blade is used HgCl ₂Soak sterilization;

(3) inoculate and induce: the blade of the Angelica keiskei after will sterilizing is cut into the square fragment of 0.5～1.0cm, is seeded on the MS medium that induced bundle sprouts, and cultivates 20d and produces green bud point, continues to cultivate 10～15d differentiation and grows up to the bud of growing thickly;

(4) take root: bud to be grown thickly grows to 2～3cm, cuts, and inserts in the MS root media that contains NAA 0.5mg/L and carries out culture of rootage.

In the above-mentioned steps (2),, use alcohol-pickled 30～60s of 70% then, use 0.1% HgCl again with flowing water flushing blade 3～5min ₂Soak sterilization 5～7 min, do not stop to rock the bubble that reduces on the explant, make blade be immersed in sufficiently sterilised in the mercuric chloride fully, use aseptic water washing at last 6～8 times.

In the above-mentioned steps (3), the MS medium that said induced bundle is sprouted consists of: in the MS medium, add 1 mg/L 2,4-D and 0.2 mg/L 6-BA.

Condition of culture is in above-mentioned steps (3) and (4) process: 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 the bud of growing thickly; Avoid first this process of generation callus, practiced thrift induction time, reduced the induced mutation rate; For the exploitation of Angelica keiskei medical value provides the raw material basis, has good application prospects.

Description of drawings

The bud point picture (20d) that Fig. 1 observes down for stereomicroscope;

The bud picture (30d) of growing thickly that Fig. 2 observes down for stereomicroscope;

Fig. 3 is for directly growing the picture of the bud of growing thickly at different leaf explants edge.

Embodiment

Embodiment 1

(1) draws materials: become to get in the strain comparatively fresh intact leaf as explant at well-grown Angelica keiskei.

(2) sterilization: with flowing water flushing blade 4 min, then on superclean bench with 70% alcohol-pickled 25 s, use 0.1% HgCl again ₂Soak sterilization 6 minutes, do not stop to rock, purpose is to reduce the bubble on the explant, and blade is immersed in the mercuric chloride fully, and sterilization is fully used aseptic water washing 7 times at last.

(3) inoculate and induce: the blade of the Angelica keiskei that disinfects is cut into 0.5～1.0 centimetre square fragment; 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, and bud point directly grows up to the bud of growing thickly (Fig. 2).Through the data acquisition statistics, 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, with inducing the indefinite bud that obtains to cut, insert in the root induction medium of MS+ NAA 1mg/L and cultivate that temperature is 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 tissue cultivating seedling after week.

Claims

1. Angelica keiskei method for quickly breeding is characterized in that: the blade that utilizes Angelica keiskei produces grow thickly bud without direct inducing culture of callus stage as explant under isolated condition, utilize inducing clumping bud to take root afterwards again and grow up to whole plant.

2. according to the said Angelica keiskei method for quickly breeding of claim 1, it is characterized in that may further comprise the steps:

(2) sterilization: blade is used HgCl ₂Soak sterilization;

3. according to the said Angelica keiskei method for quickly breeding of claim 2, it is characterized in that: in the step (2),, use alcohol-pickled 30～60s of 70% then, use 0.1% HgCl again with flowing water flushing blade 3～5min ₂Soak sterilization 5～7 min, do not stop to rock the bubble that reduces on the explant, make blade be immersed in sufficiently sterilised in the mercuric chloride fully, use aseptic water washing at last 6～8 times.

4. according to the said Angelica keiskei method for quickly breeding of claim 2, it is characterized in that: in the step (3), the MS medium that said induced bundle is sprouted consists of: in the MS medium, add 1 mg/L 2,4-D and 0.2 mg/L 6-BA.

5. the method for breeding fast according to the said Angelica keiskei of claim 2 is characterized in that: condition of culture is in step (3) and (4): intensity of illumination is 40 μ molm ^-2S ^-1, light application time 16h, temperature is 25 ℃ ± 2 ℃.