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Method for inducing generation of fritillaria cirrhosa embryoids

An embryoid body and Fritillary fritillary technology, which is applied in the field of inducing the embryoid body production of Fritillaria fritillary, can solve the problems of difficulty in ensuring the stability of the original species, long induction and culture period, and high cost, and achieves low cost and operability. Strong, low investment effect

Active Publication Date: 2013-12-25
薛刚
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This culture method not only has the problems of long induction culture period and high cost, but also has the problem that the culture material is prone to cell variation during the long-term culture process, making it difficult to ensure the stability of the original species
At present, there is no report of obtaining regenerated plants by cultivating Fritillaria fritillaria embryoids

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1: a kind of method of inducing fritillary fritillary embryoid body to produce, it comprises the following steps:

[0021] S1: Select the young leaf sheaths of Fritillaria sichuanensis plants as explants, wash them with running water, disinfect them with 70%-75% alcohol for 30-60 seconds, and then disinfect them with 0.1%-0.15% mercury chloride 4 to 10 minutes, finally shake and wash with sterile water 3 to 6 times, dry the water on the surface of the material with sterilized filter paper after washing, and inoculate the treated leaf sheath in the medium MS+6-BA 0.5 to 2.0mg· L -1 +2,4-D 1~2mg·L -1 +IAA 0.1~1mg·L -1 +20~60g·L of sucrose -1 +Agar 5.0~7.0 g·L -1 The culture was carried out in the culture medium, and the culture temperature adopted was 16-20°C, the light was 6-14 hours per day, and the light intensity was 1000-2000 lx to induce the callus;

[0022] S2: Select the white and light yellow callus treated in step S1, cut into 0.5-2.5cm in size...

Embodiment 2

[0024] Embodiment 2: a kind of method of inducing fritillary fritillary embryoid body to produce, it comprises the following steps:

[0025] S1: Select young leaf sheaths of Fritillaria sichuanensis plants as explants, wash them with running water, disinfect them with 70% alcohol for 30 seconds, then disinfect them with 0.1% mercury chloride for 4 minutes, and finally shake them with sterile water Wash 3 times, blot the water on the surface of the material with sterilized filter paper after washing, and inoculate the treated leaf sheath in medium MS+6-BA 0.5mg L -1 +2,4-D 1mg·L -1 +IAA 0.1mg·L -1 + sucrose 20g·L -1 +Agar 5.0g·L -1 The culture was carried out in , and the culture temperature adopted was 16°C, the light was 6h per day, and the light intensity was 1000lx to induce the callus;

[0026] S2: Select the white and light yellow callus treated in step S1, cut into 0.5-2.5cm in size 2 , and then insert MS+cefotaxime sodium 200mg·L -1 Cultured in medium for 40 day...

Embodiment 3

[0028] Embodiment 3: a kind of method of inducing fritillary fritillary embryoid body to produce, it comprises the following steps:

[0029] S1: Select the young leaf sheaths of Fritillaria sichuanensis plants as explants, wash them with running water, disinfect them with 75% alcohol for 60 seconds, then disinfect them with 0.15% mercury liter for 10 minutes, and finally shake them with sterile water Wash 6 times, blot the water on the surface of the material with sterilized filter paper after washing, and inoculate the treated leaf sheath in medium MS+6-BA 2.0 mg·L -1 +2,4-D 2mg·L -1 +IAA 1mg·L -1 + sucrose 60g·L -1 +Agar 7.0g·L -1 The culture was carried out in the culture medium, and the culture temperature adopted was 20°C, the light was 14h per day, and the light intensity was 2000lx to induce the callus;

[0030]S2: Select the white and light yellow callus treated in step S1, cut into 2.5cm in size 2 , and then insert MS+cefotaxime sodium 600mg·L -1 Cultured in m...

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Abstract

The invention discloses a method for inducing generation of fritillaria cirrhosa embryoids, belonging to the technical field of biologics. The method comprises the following steps of selecting tender leaf sheath of a fritillaria cirrhosa plant as an explant, sterilizing, subsequently carrying out callus induction, grafting the callus into an MS (Mass Spectrometry)+cefotaxime sodium culture medium, culturing for 50 days to grow embryoids on the surface of the callus, further cutting down the embryoids taking 1-3 embryoids as one group, grafting into a rapid growth culture medium with 0.1-1mg.L<-1> MS+6-BA+100-450mg.L<-1> cefotaxime sodium, culturing for 60 days under the condition that the culture temperature is 15-25 DEG C, the embryoids are shined for 6-20 hours every day, and the shining intensity is 800-2,500lx, so as to grow the embryoids into green small fritillaria cirrhosa plants. According to the method, the fritillaria cirrhosa embryoids can be induced by utilizing antibiotic chemical factors of a certain concentration, and a novel way is provided for breeding of the fritillaria cirrhosa embryoids.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for inducing the embryoid body of Fritillaria fringiosa to produce. Background technique [0002] Fritillaria cirrhosa D.Don is a perennial herb of Liliaceae. Its bulbs are used as medicine. It grows at an altitude of 3500m~4500m. It is mainly produced in Sichuan, Tibet, Yunnan and other provinces. Chuan Fritillaria likes cold and cool climate conditions, and has the characteristics of cold resistance, humidity, high temperature, and shade. Chuan Fritillaria is an important good medicine for treating cough. It is used for symptoms such as lung heat dry cough, dry cough with little phlegm, yin deficiency and labor cough, and expectoration with blood. Fritillary fritillary plants grow in alpine shrubs or grasslands at an altitude of 2800-4700m, and mainly rely on seeds for sexual reproduction. However, because of the morphology and physiological after-ripening cha...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 薛刚王跃华余强王晓蓉
Owner 薛刚
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