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A method for obtaining hairy roots with high anthocyanin content in plants

A high anthocyanin and hairy root technology, applied in the field of plants, can solve problems such as difficulty in construction work, sexual hybridization of hairy roots, and difficulty in obtaining hairy roots

Active Publication Date: 2016-01-06
CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Hairy roots cannot be sexually hybridized, and it is difficult to construct the overexpression of two genes in one vector, so it is difficult to obtain plant hairy roots with high anthocyanin content

Method used

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Examples

Experimental program
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Embodiment Construction

[0014] This specific embodiment adopts following technical scheme: the steps that it obtains are as follows:

[0015] (1), design primers comprising AttB1adapter and AttB2adapter, connect the MYB genes AmROS1 and AmDEL that regulate anthocyanin synthesis and metabolism in plants to the vector pDONR207 through the BP reaction of Gateway cloning; then use the LR reaction of Gateway cloning to separate AmROS1 and AmDEL Ligated into vector pJAM1502 (kanamycin resistance) and vector pK7WG2D (spectinomycin resistance) to obtain pJAM1502::AmROS1 and pK7WG2D::AmDEL;

[0016] (2), using the method of electroporation to transform PJAM1502::AmROS1 and pK7WG2D::AmDEL into Agrobacterium rhizogenes (rifampicin resistance), and select by rifampicin, kanamycin and spectinomycin antibiotics, Agrobacterium rhizogenes to obtain pJAM1502::AmROS1 and pK7WG2D::AmDEL;

[0017] (3), tobacco germinates and grows on the aseptic medium for 19 days, cuts the tobacco leaves into small squares in an ultra...

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Abstract

The invention discloses a method for obtaining plant hairy roots with high anthocyanin content, and relates to the technical field of plants. The method comprises the following steps of respectively constructing an MYB gene and a bHLH gene AmDEL for regulating anthocyanin anabolism into a carrier PJAM 1502 and a carrier pK7WG2D to obtain PJAM1502::AmROS1 and pK7WG2D::AmDEL; converting PJAM1502::AmROS1 and pK7WG2D::AmDEL into agrobacterium rhizogenes to obtain the agrobacterium rhizogenes containing plasmids PJAM1502::AmROS1 and pK7WG2D::AmDEL; and inducing out hairy roots with high anthocyanin content after utilizing the converted agrobacterium rhizogenes to infect plant tissue organs for co-culturing and screening-culturing. The method disclosed by the invention simultaneously expresses transcription factor MYB gene and bHLH gene for regulating anthocyanin anabolism to obtain the special plant hairy roots with high anthocyanin content.

Description

Technical field: [0001] The invention relates to a method for obtaining hairy roots with high anthocyanin content of plants, which belongs to the technical field of plants. Background technique: [0002] Anthocyanins are unique and ubiquitous water-soluble pigments of a class of terrestrial plants, giving plant organs red, pink, blue and purple colors. As a food, anthocyanins are not only excellent food additives, but also have anti-arrhythmia, anti-cancer functions, and reduce the incidence of cardiovascular diseases, obesity, and diabetes. [0003] Anthocyanins belong to flavonoids in phenolic compounds. The basic structural unit is the 2-phenylbenzopyran-type cation, that is, the flower color motif. The difference in the number of hydroxyl groups, methylation, glycosylation number, type and connection position of the flower color motif forms more than 200 different compounds. These anthocyanins with different structures will show different colors and have different hea...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82A01H5/06
Inventor 刘宝龙张怀刚刘登才张波陈文杰沈裕虎魏乐
Owner CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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