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Induction culture method of bletilla striata callus

A callus induction and callus technology, applied in horticultural methods, botanical equipment and methods, gardening, etc., can solve the problems of cumbersome, unfavorable adjustment, easy to damage callus, etc., and achieve the effect of simplifying the cultivation steps

Pending Publication Date: 2019-06-14
ZUNYI MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above operations are very cumbersome. The process of taking out the callus from the solid induction medium and subsequent treatment is easy to damage the callus. Because the callus has a growth core when it grows, once it is destroyed, it is difficult for the callus to resume division and growth.
Moreover, the choice of solid proliferation medium is not conducive to the formation of well-dispersed free small cell clusters by callus, and it is difficult to provide callus with uniform properties for subsequent induction and differentiation culture.
In addition, the use of solid induction medium and subculture medium is not conducive to the adjustment and control of the levels of various nutrients and hormones in the medium, and is not conducive to the optimization and improvement of the induction culture method and process.

Method used

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  • Induction culture method of bletilla striata callus
  • Induction culture method of bletilla striata callus
  • Induction culture method of bletilla striata callus

Examples

Experimental program
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Effect test

Embodiment 1

[0047] Step 1. Take uncracked Baiji and mature capsules, rinse them under running water for 20 minutes, peel off Baiji capsules, and pour Baiji seeds into a sterilized 10ml EP tube. Put the EP tube in the ultra-clean workbench and open it, soak it with 75% alcohol for 30s, and shake it constantly, then rinse the seeds with sterile water for 3 times, then add 0.1% Tween-80 containing two drops to the EP tube Mercury chloride solution, soak the seeds for 20s, shake constantly during the soaking process, and finally rinse the seeds with sterile water for 5 times, dry the moisture on the surface of the seeds with absorbent paper, and set aside;

[0048] Step 2, transfer the obtained sterile white and seeds into the induction medium, the pH value of the induction medium is 6.0, the induction culture temperature is 25°C, and the way of dark culture is adopted, and the rotation speed is 120r / min. The induction medium consists of the following components: MS liquid medium, 0.480mg / L 6...

Embodiment 2

[0052] The operation method of this example is basically the same as that of Example 1, the difference is that in Step 2 and Step 3, the induction medium and subculture medium used are composed of the following components: MS liquid medium, 0.760 mg / L 6-BA, 1.635mg / L 2,4-D, 0.631mg / L NAA, 30g / L sucrose.

[0053] In this example, a large amount of callus appeared in the induction medium after 32 days of induction culture, and the induction rate of callus in this example was 91.69%. After 20 days of subculture (see figure 2 ), observe that the callus obviously expands than the callus before 20 days after subculture in suspension culture, the structure is more loose, the callus is rarely browned, the medium is clear, and the calculated callus proliferation rate is 6.28%.

Embodiment 3

[0055] The operation method of this example is basically the same as that of Example 1, except that in Step 2 and Step 3, the induction medium and subculture medium used are composed of the following components: MS liquid medium, 1.000 mg / L 6-BA, 2.000mg / L 2,4-D, 1.000mg / L NAA, 20g / L sucrose.

[0056] In this example, a large amount of callus appeared in the induction medium after 36 days of induction culture, and the induction rate of callus in this example was 90.90%. After 20 days of subculture, it was observed that the callus after suspension culture was significantly larger than the callus before 20 days, the structure was more loose, the callus was rarely browned, the medium was clear, and the calculated callus proliferation The rate is 6.17%.

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Abstract

The invention relates to the technical field of traditional Chinese medicine research, and relates to a callus culture method, in particular to an induction culture method of bletilla striata callus,which comprises callus induction culture of bletilla striata seeds and subculture of callus, wherein both the callus induction culture and the subculture adopt a suspension culture mode, and an MS liquid culture medium containing a plant growth regulator is used. The plant growth regulator is 6- benzylaminopurine, 2, 4-dichlorophenoxyacetic acid and naphthaleneacetic acid. The induction culture method of bletilla striata callus can facilitate the process of researching the optimal growth and differentiation conditions of the bletilla striata callus, reduce the influence of toxic secondary metabolites on the white and the callus, and provide a relatively consistent bletilla striata callus cell group and the like. The method can be applied to industrial production of secondary metabolites produced by suspension culture of bletilla striata callus.

Description

technical field [0001] The invention relates to the technical field of research on traditional Chinese medicinal materials, in particular to a callus culture method, in particular to a callus induction culture method. Background technique [0002] Bletilla striata (Thunb.) Rchb.f. is a perennial herbal medicinal plant of the genus Bletilla striata in the family Orchidaceae. It is mainly produced in the Yangtze River Basin, Gansu, Shaanxi and other places in my country. Baiji is a traditional Chinese medicinal material with extensive medicinal value. The pharmacopoeias of the past dynasties recorded that it has the effects of astringent and hemostasis, detumescence and granulation, and can be used to treat hemoptysis, hematemesis, traumatic bleeding, sore swelling, chapped skin and other diseases. Modern pharmacological studies have shown that Baiji has antibacterial, anti-oxidant, anti-tumor, anti-peptic ulcer, and immune regulation effects. In addition to its medicinal ap...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 徐德林上官艳妮李树刚何毅怀李林潘胤池刘厚伯李杰
Owner ZUNYI MEDICAL UNIVERSITY
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