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Method for cultivating bletilla striata seedlings by using intermittent immersion bioreactor

A technology of bioreactor and bletilla striata, which is applied in the field of cultivating bletilla striata seedlings using intermittent immersion bioreactors, can solve the problems of unreported and public application of bletilla striata seedlings, shorten the cultivation period, reduce the pollution rate, and improve the automation level Effect

Active Publication Date: 2018-01-12
NANJING TECH UNIV
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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 cultivating bletilla striata seedlings by using intermittent immersion bioreactor
  • Method for cultivating bletilla striata seedlings by using intermittent immersion bioreactor

Examples

Experimental program
Comparison scheme
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 th...

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, specifically relates to the technical field of medicinal plant cultivation, and in particular to a method for cultivating Bletilla striata seedlings using an intermittent immersion bioreactor; the method for cultivating Bletilla striata seedlings using an intermittent immersion bioreactor, Including seed sterilization, seed germination, multiplication and cultivation, pseudobulb generation and cultivation and pseudobulb expansion and cultivation steps, the method for rapidly cultivating Bletilla striata seedlings using intermittent immersion bioreactors is carried out under aseptic conditions, and the multiplication and cultivation includes transition Proliferation culture and Proliferation and rooting culture. Through the above technical scheme, the proliferation culture process is divided into two processes of transitional proliferation culture and proliferation and rooting culture, and the transitional proliferation culture eliminates or shortens the long adaptation period from solid culture to intermittent immersion bioreactor culture by means of shake flasks phenomenon, thereby rapidly obtaining a large amount of bletilla striata seedlings for production, and then obtaining the method for the pseudobulb seedlings through the cultivation of pseudobulb expansion.

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, its d...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 陈集双宋莲张本厚蒋海侠
Owner NANJING TECH UNIV
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