A method for controlling endophytic bacteria contamination in tissue culture of Taibai Fritillaria

A technology of taibai fritillary and tissue culture, applied in the biological field, can solve problems such as low proportion and infection of endophytes, and achieve the effects of reducing production cost, controlling pollution rate, and fast bulb reproduction and growth.

Active Publication Date: 2019-06-07
SICHUAN ACAD OF CHINESE MEDICINE SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the prior art, the bulbs cultivated by this method have a very high probability of infecting endophytes during the cultivation process, and the ratio of successfully carrying out the transfer and continuing cultivation is low.

Method used

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  • A method for controlling endophytic bacteria contamination in tissue culture of Taibai Fritillaria
  • A method for controlling endophytic bacteria contamination in tissue culture of Taibai Fritillaria
  • A method for controlling endophytic bacteria contamination in tissue culture of Taibai Fritillaria

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Embodiment 1

[0028] 1. Explant selection and pretreatment:

[0029] Excavate the fresh bulbs of Taibai Fritillaria from March to April as explants, remove the rotten and browned scales on the outer layer, and cut off the root system; wash the soil on the surface of the bulbs, soak them in a washing powder solution with a concentration of 1g / L for 5 minutes, Scrub the surface gently with a brush, rinse with running water for 1 hour, then rinse with sterile deionized water for 5 times, blot dry with filter paper, and set aside;

[0030] 2. Disinfection treatment of explants: put the treated explants into a jar, and pour 0.1% mercuric chloride solution at normal temperature until the explants are submerged. Stir the explants with a magnetic stirrer at a low speed for 6-8 minutes, wipe the surface of the jar with 75% alcohol and put it in an ultra-clean workbench. Under aseptic conditions, rinse the bulbs with sterile water for 3-5 times, 3-5 minutes each time , and finally dry it with steril...

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Abstract

The invention relates to the field of biotechnology, in particular to a method for controlling endophytic bacteria contamination in tissue culture of Taibai Fritillaria. The control method of the present invention comprises: S1, explant selection and pretreatment: select the fresh bulb of Taibai Fritillaria, soak in washing powder solution, wipe, rinse, filter paper blot, standby; S2, explant disinfection treatment: pretreatment The final explants were submerged in mercuric chloride solution, stirred, placed on a clean bench, rinsed, dried, and set aside; S3, small bulb induction differentiation: split the explant scales obtained by S2 treatment, and cut them longitudinally into 2 ~4mm wide small pieces, inoculated on the culture medium added with penicillin and carbendazim; S4, subculture: subculture once every 20 days; S5 small bulb proliferation culture: transfer the healthy small bulbs obtained in S4 cultured on medium without the addition of any sterilant. The method for controlling endophytic bacteria pollution in tissue culture of Taibai Fritillaria of the present invention has a control rate of endophytic bacteria pollution of up to 96.5%.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for controlling endophyte pollution in the tissue culture process of Fritillaria taibai. Background technique [0002] Fritillaria delavayi Franch. Fritillaria przewalskii Maxim. Fritillaria delavayi Franch. Dry bulbs of Fritillaria taipaiensis P.Y.Li or Fritillaria unibracteata Hsiao et K.C.Hsia var wabuensis (Y.Tang et S.C.Yue) Z.D.Liu, S.C.Chen. It is one of the bulk medicinal materials produced in precious Sichuan. It is mainly used for diseases such as lung heat and dry cough, dry cough with little phlegm, labor cough due to yin deficiency, and bloody sputum. The clinical demand is huge. [0003] Taibai Fritillaria was included in the 2010 edition of "Chinese Pharmacopoeia" as a new source. It has the characteristics of less wild resources, strong adaptability, and normal growth and development at low altitudes. It is the best variety suitable for home cultivation amon...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 胡平夏燕莉周先建杨玉霞陈铁柱方清茂郭俊霞吴萍李青苗舒光明
Owner SICHUAN ACAD OF CHINESE MEDICINE SCI
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