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Method for increasing geniposide content in Gardenia jasminoides cells

A technology of geniposide and cells, applied in the biological field, can solve problems such as late start of research

Active Publication Date: 2022-03-01
CHANGSHA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

As early as 1989, Japanese scientists carried out the research on the production of secondary metabolites by gardenia tissue culture, but domestic research on the primary metabolites and secondary metabolites obtained by Gardenia callus started relatively late
There is no report on the production of geniposide by cell culture

Method used

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  • Method for increasing geniposide content in Gardenia jasminoides cells
  • Method for increasing geniposide content in Gardenia jasminoides cells
  • Method for increasing geniposide content in Gardenia jasminoides cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Production of geniposide by large-scale culture of isolated cells of Gardenia jasminoides

[0026] 1. Induced polyploid cells

[0027] In mid-August, fresh immature gardenia fruits were picked and removed to get the seeds and disinfected. The seed coats of the sterilized seeds were removed through aseptic operation, and the peeled gardenia kernels were cut in half by a cross-section with a scalpel. Soak in 0.15% colchicine solution for 8 hours, take it out and rinse it with sterile water for more than five minutes before use;

[0028] 2. Induction of polyploid callus

[0029] Inoculating the gardenia kernels treated in step 1) on a dedifferentiation medium, and culturing them at room temperature at 25°C for 10 days to obtain callus containing polyploid cells;

[0030] Wherein, the dedifferentiation medium is a medium obtained by adding 0.5 mg / L 2,4-D, 0.25 mg / L 6-BA, 1.0 mg / L coenzyme A, and 30 mg / L sucrose to MS medium.

[0031] 3. Subculture

[0032] The...

Embodiment 2

[0037] Example 2 Production of geniposide by large-scale culture of in vitro cells from Gardenia jasminoides

[0038] 1. Induced polyploid cells

[0039] Harvest immature fresh gardenia fruits in mid-August for disinfection treatment, soak them in 0.5% colchicine solution for 7 days, remove the fruit and get their seeds through aseptic operation, peel off the seed coat and peel the peeled gardenia kernels with a scalpel Cut in half according to the cross section and set aside.

[0040] 2. Induction of polyploid callus

[0041] inoculating the gardenia kernels treated in step 1) on the dedifferentiation medium, and culturing at room temperature at about 25°C for 10 days to obtain callus containing polyploid cells;

[0042] Wherein the dedifferentiation medium is a medium obtained by adding 0.5mg / L of 2,4-D, 0.25mg / L6-BA, 2.0mg / L of coenzyme A, and 40mg / L of sucrose in MS medium;

[0043] 3. Subculture

[0044] The polyploid cell line obtained in step 2) is inoculated in MS ...

Embodiment 3

[0049] Determination of the content of geniposide in the callus of Gardenia jasminoides in embodiment 3

[0050] The content of geniposide in the cultures of Example 1 and Implementation 2 was determined by high performance liquid chromatography (HPLC), and compared with the content of geniposide in Gardenia jasminoides, the results are shown in the table below.

[0051] Table 1 Control Experiment Results (n=3)

[0052]

[0053] After the four samples were cultured by the method provided by the present invention, the content of geniposide in the obtained tissues all increased significantly, wherein samples I-IV had a higher increase rate according to the method of Example 1, and samples No. IV were according to the method of Example 2. method is higher.

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Abstract

Gardenia contains many kinds of biologically active substances. The active ingredients of gardenia are recognized at home and abroad as iridoid substances, of which the highest content is geniposide, also known as geniposide. Since gardenia is the only natural source of gardeniaside from gardenia fruit, the acquisition of gardeniaside is completely dependent on gardenia fruit. Screening gardenia-producing cells through plant cell engineering for large-scale culture is an effective way to solve the shortage of gardenia sources. one of the ways. So far, there is no report on the production of geniposide by cell culture. The invention effectively resolves the contradiction between the growth of gardenia cells and the decrease of the content of gardeniaside, and can obtain fresh cells rich in gardeniaside in a short period of time, and the content of gardeniaside in the cultured cells is the same as that of gardenia in the seed cells. Several times the content of glycosides, and is not affected and limited by natural conditions.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a method for increasing the content of geniposide in gardenia kernel cells by adopting plant cell culture technology. Background technique [0002] Gardenia is the dry and mature fruit of Gardenia jasmi-noides Ellis, a Rubiaceae plant. It has the functions of purging fire and irritability, clearing heat and promoting dampness, cooling blood and detoxification, reducing swelling and relieving pain. Geniposide is the secondary metabolite of Gardenia, Its chemical structure is the glucoside of iridoids. Gardenia contains more than 40 kinds of biologically active substances, mainly iridoids and volatile oils, and the active ingredients of Gardenia recognized at home and abroad are iridoids, of which the highest content is geniposide, also known as It is geniposide, and its aglycon is genipin. [0003] Zhu Wenpei et al. used gardenia fruits of different maturity as s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04C12P17/16
CPCC12N5/04C12P17/162
Inventor 陈建荣刘芳唐伟卓唐映红赵晨曦
Owner CHANGSHA UNIVERSITY
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