Method for cultivating tobacco rich in anthocyanin and application thereof

An anthocyanin and tobacco technology, applied in the fields of plant molecular biotechnology and genetic engineering, to reduce health hazards

Active Publication Date: 2013-04-17
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most smokers are so addicted to smoking that they cannot avoid smoking-related diseases by quitting

Method used

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  • Method for cultivating tobacco rich in anthocyanin and application thereof
  • Method for cultivating tobacco rich in anthocyanin and application thereof
  • Method for cultivating tobacco rich in anthocyanin and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Gene function prediction

[0026] (1) Experimental method

[0027] 1. Carrier construction

[0028] First construct an expression vector containing a single gene. According to confirmed MrMYB1 (SEQ: NO. 1) and MrbHLH1 (SEQ: NO. 2) sequence full-length designed primers containing restriction endonuclease sites SEQ: NO. 3 and SEQ: NO. 4 and SEQ: NO. 5 and SEQ: NO. 6, PCR amplification The open reading frames of the two genes were amplified, and then the amplified products and the expression plasmid pGreenII SK were treated with corresponding endonucleases, and finally the corresponding PCR products and plasmids were connected to contain MrMYB1 or MrbHLH1 Construction of expression vectors.

[0029] Then build with MrMYB1 and MrbHLH1 Bivalent expression vector: According to the requirements of fusion PCR technology for primers and the sequence information of expression plasmid pGreenII SK (SEQ: NO. 7), design primers SEQ: NO. 8 and SEQ: NO. 9 (amplify...

Embodiment 2

[0037] Embodiment 2: Tobacco transgenic technology

[0038] (1) Experimental method

[0039] 1. Agrobacterium liquid preparation

[0040] will contain MrMYB1 - MrbHLH1 The GV3101::pSoup Agrobacterium glycerol strain of the bivalent expression plasmid was streaked onto LB solid medium containing 25 μg / ml gentamicin, 2.5 μg / ml tetracycline and 50 μg / ml kanamycin, 28 o C cultured for 48 h; pick a small amount of colony and spread it on a new LB solid medium containing 25 μg / ml gentamicin, 5 μg / ml tetracycline and 50 μg / ml kanamycin, 28 o C culture for 24 h; scrape a few well-growing colonies, and culture them with 30 ml LB liquid containing 25 μg / ml gentamycin, 5 μg / ml tetracycline and 50 μg / ml kanamycin based on 28 o Cultivate at 100 rpm for 24 h; collect colonies by centrifugation at room temperature and 5000 rpm, suspend with 10 – 20 ml of tobacco transgenic liquid (MS + 2% sucrose + NV, pH=6) and set aside for later use.

[0041] 2. Preparation of sterile tobacco tis...

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Abstract

The invention provides a method for cultivating tobacco rich in anthocyanin and an application thereof. The method comprises the following steps of recombining two confirmed genes, namely MrMYB1 and MrbHLH1 which can be used for regulating and controlling anthocyanin synthesis in red bayberry fruits, establishing an expression plasmid, screening genes capable of regulating and controlling anthocyanin synthesis by using a tobacco leaf transient expression technique, converting genes with confirmed functions into the tobacco, and cultivating tobacco rich in anthocyanin. The invention provides a method for cultivating tobacco rich in anthocyanin, so that health hazard caused by smoking is alleviated; and the method is also applicable to MYB and bHLH genes in other fruits besides red bayberry.

Description

technical field [0001] The invention belongs to the fields of plant molecular biotechnology and genetic engineering, and relates to a method for cultivating anthocyanin-rich tobacco and its application. Background technique [0002] Smoking seriously affects human health. Smoking can not only induce cardiovascular disease, chronic lung disease, and cancer, but also increase the incidence of infectious diseases, directly leading to an increase in the incidence of severe influenza, invasive pneumonia, and tuberculosis; causing numerous reproductive complications, including infertility, Spontaneous miscarriage, low baby weight and premature death and other syndromes; make the body produce insulin resistance, which is an important factor in the onset of diabetes, and will accelerate the process of vascular disease in smokers; it will also affect the recovery of the body after surgery, including delaying Wound healing and increase the chance of wound infection, etc. [0003] Wi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84A01H5/00
Inventor 孙崇德徐昌杰殷学仁刘晓芬李鲜张波陈昆松
Owner ZHEJIANG UNIV
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