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Application of plant NIA1 gene in increasing content of flavone and content of lactone of ginkgoes

A gene and plant technology, applied in the fields of plant genetic improvement, botanical equipment and methods, and applications, can solve problems such as rarely seen research reports, and achieve the effect of increasing the content of total flavonoids and total ginkgolides

Inactive Publication Date: 2016-03-02
HANGZHOU NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the lack of molecular means for compound regulation of secondary metabolism at the level of the cell as a whole, there are few relevant research reports so far.

Method used

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  • Application of plant NIA1 gene in increasing content of flavone and content of lactone of ginkgoes
  • Application of plant NIA1 gene in increasing content of flavone and content of lactone of ginkgoes
  • Application of plant NIA1 gene in increasing content of flavone and content of lactone of ginkgoes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Construction of transgenic Ginkgo cell line with NIA1 gene

[0026] (1) NIA1 gene: Using the Arabidopsis NIA1 gene sequence in public databases such as GeneBank as a template, amplify the NIA1 gene from Arabidopsis or entrust a professional bioengineering technology company (Shanghai Sangon Bioengineering Technology Service Co., Ltd.) to synthesize it Arabidopsis NIA1 gene (shown in SEQ ID NO.1).

[0027] (2) Digest the NIA1 gene with EcoRI+BamHI, recover the digested fragment, and ligate it with the large fragment of the plasmid pBIN19 (purchased from NTCC Type Culture Collection Center) digested with EcoRI+BamHI at 16°C overnight to obtain a new plasmid pBIN19 -NIA1.

[0028] The reaction system is as follows:

[0029] Table 1 pBIN19, NIA1 ligation reaction system

[0030]

[0031] (3) Plasmid pCHS (purchased from NTCC Type Culture Collection Center) was digested with HindⅢ+XbaⅠ, and the small fragment containing the CaMV35S promoter was recovered, an...

Embodiment 2

[0042] Example 2: Promotion of NIA1 gene to the synthesis of flavonoids and ginkgolides in ginkgo cells

[0043] (1) Using the NIA1 gene transgenic ginkgo suspension cell line obtained in Example 1 as material, inoculate it with 0.5mg / LNAA, 1.0mg / L6-BA, 0.1mg / L2, 4-D, 30g at a ratio of 0.05mg / ml In the MS liquid medium (without agar) of / L sucrose, shake culture (140rpm) in the dark under 28 ℃;

[0044] (2) Using non-genetically transformed ginkgo cells as materials, carry out suspension subculture according to the steps in Example 1 of the above-mentioned method to obtain the wild-type suspension cell line of Ginkgo biloba, and carry out according to the same culture conditions as step (1). to cultivate;

[0045] (3) Sampling at different periods of cell growth, and measuring the total flavonoid content according to the HPLC method reported in the literature (Wang Jing; Zou Yujia; Tang Huacheng; Li Dong. High performance liquid chromatography (HPLC) measures the content of g...

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Abstract

The invention discloses application of a plant NIA1 gene in increasing the content of flavone and the content of lactone of ginkgoes. The NIA1 gene of arabidopsis is transferred into gingko cells and stably expressed through an agrobacterium tumefaciens mediated transformation method, after culturing is carried out for 35 days, the total flavone content and total lactone content in NIA1-modified gingko cells are increased by 103.5% and 78.4% respectively compared with those in non-genetically-modified gingko cells, and thus significant application meaning is achieved on simultaneously increasing the content of flavone and the content of lactone in gingkoes.

Description

(1) Technical field [0001] The invention relates to an application of a plant NIA1 gene, in particular to an application of a plant NIA1 gene to simultaneously promote the synthesis and accumulation of flavonoids and ginkgolides in ginkgo cells. (2) Background technology [0002] Ginko Biloba Extract (GBE) is a product obtained by extracting the active ingredients from Ginkgo biloba leaves as raw material with an appropriate solvent. GBE is widely used in pharmaceuticals, health products, food additives, functional beverages, cosmetics and other fields. This product is one of the most successful cases of herbal medicine developed by modern science and technology. [0003] The medicinal value and application of Ginkgo biloba extract are extremely extensive. In addition to significantly antagonizing PAF receptors, it can also play a role in anti-inflammation, anti-allergy, dilating blood vessels, protecting cardiovascular and cerebrovascular, improving peripheral blood circu...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/84C12N5/10
Inventor 徐茂军蒋智芳杨燕君许名峰叶昱郑耀奇邱蓉蓉
Owner HANGZHOU NORMAL UNIVERSITY
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