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Method for increasing content of vallesiachotamine in suspension cells of Catharanthus roseus by using mixed elicitor of methyl jasmonate and silver thiosulfate

A technology of methyl jasmonate and silver thiosulfate, applied in the field of bioengineering, can solve the problems of cell contamination, time-consuming replacement of culture medium, inconvenient application, etc., and achieves the effects of easy operation and high content

Inactive Publication Date: 2016-11-23
于荣敏
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  • Application Information

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Problems solved by technology

However, the two-step method is inconvenient to apply in industrial production, and it is time-consuming and easy to cause cell contamination when changing the medium

Method used

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  • Method for increasing content of vallesiachotamine in suspension cells of Catharanthus roseus by using mixed elicitor of methyl jasmonate and silver thiosulfate
  • Method for increasing content of vallesiachotamine in suspension cells of Catharanthus roseus by using mixed elicitor of methyl jasmonate and silver thiosulfate
  • Method for increasing content of vallesiachotamine in suspension cells of Catharanthus roseus by using mixed elicitor of methyl jasmonate and silver thiosulfate

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Experimental program
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Embodiment Construction

[0009] Experimental Materials

[0010] Periwinkle suspension cell culture system.

[0011] Preparation of culture medium

[0012] Prepare B5+IAA according to conventional methods 0.5 +6-BA 1.0 Medium.

[0013] Preparation of elicitor

[0014] Put 0.924 g Na 2 S 2 o 3 ·5H 2 O was dissolved in 50 ml deionized water, and then 0.16 g AgNO 3 Dissolve in 50 ml deionized water. Then put AgNO 3 The solution was slowly poured into the sodium thiosulfate solution, and stirred rapidly while pouring to obtain a silver thiosulfate solution with a concentration of 9.2 μmol / L. Weigh 39.2 mg of methyl jasmonate and dissolve it in 10 ml of absolute ethanol to obtain a solution of methyl jasmonate with a concentration of 0.018 μmol / L.

[0015] training method

[0016] Each implementation is in B5+IAA respectively 0.5 +6-BA 1.0 In the culture medium, cultivated under light conditions for 12 days, and added different amounts of elicitors according to different concentration require...

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Abstract

The invention relates to a method for promoting synthesis of vallesiachotamine from suspension cells of Catharanthus roseus through synergism of a mixed elicitor of methyl jasmonate and silver thiosulfate. The cells of Catharanthus roseus is subjected to illumination culture in a B5+IAA<0.5>+6-BA<1.0> medium at a temperature of 25 DEG C for 12 days; then the mixed elicitor of methyl jasmonate and silver thiosulfate is added; culture is continued under same culture conditions for 1 to 7 days, and then harvesting is carried out; and alkaloids are extracted from a culture product, and changes of the content of vallesiachotamine are detected by using a HPLC method. Results show that after addition of the mixed elicitor of methyl jasmonate and silver thiosulfate, generation of vallesiachotamine by secondary metabolites in the suspension cells of Catharanthus roseus is substantially promoted; and the content of vallesiachotamine reaches a maximal value in 5 days, being 75.2 times of the content of a control group. The induction action of the mixed elicitor is better than the induction action of an individual elicitor; and it is showed that the mixed elicitor of methyl jasmonate and silver thiosulfate can cooperatively promote synthesis of vallesiachotamine by the secondary metabolites in the cells of Catharanthus roseus.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a method for increasing the yield of syringine by adding mixed elicitor methyl jasmonate and silver thiosulfate to the suspension cell culture medium of periwinkle flowers. Background technique [0002] Periwinkle( Catharanthus roseus ) is an important medicinal plant of the Apocynaceae family, native to the Mediterranean coast, India, and tropical America. The history of China's cultivation of periwinkle is not long. It is mainly cultivated in the south of the Yangtze River, and it is more common in provinces (autonomous regions) such as Guangdong, Guangxi, and Yunnan. Vinca plants contain more than 130 alkaloids, most of which have pharmacological activity, vinblastine and vincristine are currently one of the most widely used natural plant anti-tumor drugs; serpentine (serpentine) ) can be used to treat diseases such as hypertension and arrhythmia; ajmalicine has a sedat...

Claims

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Application Information

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IPC IPC(8): A01H1/00
Inventor 于荣敏刘金伟张文锦
Owner 于荣敏
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