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Method for culturing bletilla striata seedling by virtue of intermittent immersion type bioreactor

A technology of bioreactor and bletilla striata, which is applied in the field of cultivating bletilla striata seedlings by using intermittent immersion bioreactors, can solve the problems of unreported and public application of bletilla striata seedlings, shorten the cultivation period, improve the automation level, and improve plant proliferation efficiency effect

Active Publication Date: 2015-06-24
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In view of utilizing intermittent immersion culture method to carry out the production of Bletilla striata seedlings and there is no report and open application, therefore, it is necessary to develop a kind of method utilizing intermittent immersion bioreactor to cultivate Bletilla striata seedlings

Method used

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  • Method for culturing bletilla striata seedling by virtue of intermittent immersion type bioreactor
  • Method for culturing bletilla striata seedling by virtue of intermittent immersion type bioreactor

Examples

Experimental program
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Effect test

Embodiment 1

[0049] Embodiment 1: as figure 1 as shown,

[0050] (1) Pre-preparation steps: Sterilization of reactors and transparent containers: bandage the reactors and transparent containers, sterilize them with high temperature and high pressure and moist heat for later use; the reactor is a gap immersion bioreactor, and the transparent container is an Erlenmeyer flask;

[0051] (2) Seed sterilization step: perform aseptic treatment on fine seed pods of Bletilla striata;

[0052] Surface decontamination: rinse with tap water for 30 minutes; surface disinfection: then soak the cleaned Bletilla striata seed pods in 70~75% (v / v) alcohol for 30 (s) in an ultra-clean workbench, and then soak them in sterile water Rinse 2-3 times; surface sterilization: soak in mercuric chloride with a concentration of 0.1% (g / v) for 8 minutes, rinse 5-7 times with sterile water; Operation Cut a small mouth from one end of the bletilla striata seed pod;

[0053] (3) Seed germination steps: According to t...

Embodiment 2

[0061] Utilize intermittent immersion bioreactor to cultivate bletilla striata seedlings, concrete steps are as shown in embodiment 1, what differ with it is that semi-solid culture is adopted during transition multiplication culture, and the selected medium formula is 1 / 2MS+NAA. .5mg / L+50g / L potato liquid+30g / L sucrose+agar 3g / L, adjust the pH to 5.9, the formula of proliferation and rooting culture medium is 1 / 2MS+NAA1.0mg / L+50g / L potato liquid+30g / L sucrose, adjust the pH to 5.9, the culture conditions are light intensity 1500~2000lx, light time 12h / d, temperature 24±1℃. In one culture cycle, the adaptation period of Bletilla striata seedlings in the intermittent immersion bioreactor was shortened to 0 days, so that the proliferation efficiency of the reactor reached more than 1:30.

Embodiment 3

[0063] Utilize the bioreactor to cultivate Bletilla striata seedlings, the specific steps are as shown in Example 1, the difference is that it is a semi-solid culture that is adopted during the transitional multiplication culture, and the selected medium formula is 1 / 2MS+NAA0.5mg / L+50g / L potato liquid+30g / L sucrose+agar 2g / L, adjust the pH to 6.0, the culture medium for pseudobulb expansion is MS+NAA1.0mg / L+50g / L potato liquid+banana juice 40g / L+straw Juice 60g / L, adjust pH6.0, the adaptation period of Bletilla striata seedlings in the intermittent immersion bioreactor is shortened to 0.3 days, in a culture period, the pseudobulbs of Bletilla striata seedlings expand about 2 ~ 3 times, and the seedling survival rate obtained Above 90%.

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Abstract

The invention belongs to the field of traditional Chinese medicinal materials and particularly relates to a method for culturing a bletilla striata seedling by virtue of an intermittent immersion type bioreactor. The method sequentially comprises the steps the following steps: seed sterilization, seed germination, enrichment culture, pseudobulb generation culture and pseudobulb expansion culture. The method for culturing the bletilla striata seedling by virtue of the intermittent immersion type bioreactor is carried out under aseptic conditions; the enrichment culture step comprises a transition enrichment culture step and a multiplication and rooting culture step. By virtue of the technical scheme, the enrichment culture process is divided into the transition enrichment culture process and the multiplication and rooting culture process, and in the transition enrichment culture process, a long adaptation phenomenon occurring from a solid culture process to an intermittent immersion type bioreactor culture process is eliminated or shortened by virtue of a bottle shaking manner, so that lots of bletilla striata seedlings for production are rapidly obtained and then are subjected to pseudobulb expansion culture to produce seedlings with pseudobulbs.

Description

technical field [0001] The invention belongs to the technical field of medicinal plant cultivation, and in particular relates to a method for cultivating bletilla striata seedlings by using an intermittent immersion bioreactor. Background technique [0002] Bletilla striata (Bletilla striata (Thunb.) Reichb.f.) is a perennial herbaceous plant of the genus Bletilla striata in the family Orchidaceae, also known as "Bletilla striata", Lianji grass, etc. Its dried tubers have the effects of astringent and hemostasis, detumescence and muscle growth, and are often used to treat hemoptysis, hematemesis, and chapped skin. Studies have shown that Bletilla striata has antibacterial and antitumor effects. Bletilla striata is not only used in clinical Chinese medicine, but also widely used in daily chemical products, such as cosmetics, advanced ceramics manufacturing, textiles, rubber production, etc. [0003] With the continuous expansion of the application of bletilla striata, i...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 陈集双宋莲张本厚蒋海侠
Owner NANJING UNIV OF TECH
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