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Transgenic method for improving salvianolic acid B content in root of red-rooted salvia

A salvianolic acid and transgenic technology, applied in genetic engineering, plant genetic improvement, botanical equipment and methods, etc., can solve the problem that a single genetic modification is difficult to work

Inactive Publication Date: 2013-07-10
SHAANXI NORMAL UNIV
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  • Abstract
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  • Claims
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Problems solved by technology

[0004] The plant metabolic pathway is a multi-step reaction involving a variety of enzymes, which is affected by factors such as development and environment, and it is sometimes difficult to modify a single gene

Method used

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  • Transgenic method for improving salvianolic acid B content in root of red-rooted salvia
  • Transgenic method for improving salvianolic acid B content in root of red-rooted salvia
  • Transgenic method for improving salvianolic acid B content in root of red-rooted salvia

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Embodiment 1

[0028] The experimental method that does not indicate specific condition in this embodiment, operates according to known method, such as Sambrook etc. " molecular cloning: laboratory handbook " (New York: Cold Spring Harbor Laboratory Press, 1989) described condition Or operate under the conditions recommended by the manufacturer.

[0029] 1. Cloning of full-length cDNA of Arabidopsis anthocyanin production transcription factor 1 gene

[0030] (1) Extraction of Arabidopsis total RNA

[0031]Select fresh and robust whole plants of Arabidopsis thaliana seedlings, and use the Biozol total RNA extraction kit produced by Hangzhou Biozol Technology Co., Ltd. to isolate and extract their total RNA. The extraction method refers to the Biozol Total RNA Extraction Kit (Cat. # BSC51S1) instructions.

[0032] (a) Preparatory work for extracting total RNA from Arabidopsis thaliana

[0033] All kinds of glass instruments, gun tips, and centrifuge tubes used were washed, soaked in a 0.1...

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Abstract

The invention discloses a transgenic method for improving salvianolic acid B content in root of red-rooted salvia, which comprises the following steps of: obtaining cDNA overall length of arabidopsis anthocyanidin generation transcription factor 1 gene through gene cloning; constructing a plant expression vector containing the genes; transforming agrobactrium tumefaciens EHA105 by using the vector so as to obtain an agrobactrium tumefaciens strain containing the genes; transforming the root of red-rooted salvia by using the agrobactrium tumefaciens strain, and culturing to obtain a transgenicplant of the root of red-rooted salvia detected through a polymerase chain reaction; detecting the expression of the arabidopsis anthocyanidin generation transcription factor 1 gene in the transgenicroot of red-rooted salvia through a semi-quantitative reverse transcription-polymerase chain reaction; and performing high performance liquid chromatography on the salvianolic acid B content in the transgenic root of red-rooted salvia subjected to the gene expression, and screening to obtain a transgenic plant of the root of red-rooted salvia with the improved salvianolic acid B content. Due to the obtained transgenic plant of the root of red-rooted salvia, the salvianolic acid B content is 73.27mg / g in dry roots when the transgenic plant of the root of red-rooted salvia grows for 165 days, wherein the salvianolic acid B content is two times of the salvianolic acid B content in non-transformed common dry roots of the root of red-rooted salvia.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering of medicinal plants, and in particular relates to a method for transforming Arabidopsis thaliana anthocyanin production transcription factor 1 gene in salvia miltiorrhiza to increase the content of salvianolic acid B. Background technique [0002] Salvia miltiorrhiza (Salvia miltiorrhiza Bunge) is the dry root and rhizome of Salvia miltiorrhiza Bunge, a commonly used traditional medicinal material in my country. It has significant clinical therapeutic effects on cardiovascular and cerebrovascular diseases, cancer and various inflammations. The main active ingredients of Danshen are divided into two categories: one is fat-soluble tanshinone compounds, and the other is water-soluble phenolic acids, mainly including danshensu, rosmarinic acid and salvianolic acid B. The water-soluble compounds of Salvia miltiorrhiza have attracted more and more people's attention because of their consiste...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/82C12Q1/68G01N30/89A01H5/00
Inventor 王喆之张媛
Owner SHAANXI NORMAL UNIV
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