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Method for establishing agrobacterium rhizogene mediated peanut root inducing system, and application thereof

A technique for establishing Agrobacterium rhizogenes and a method, which is applied in the field of establishment of a peanut root induction system, and can solve the problems of not transforming foreign genes through Agrobacterium rhizogenes, infecting Agrobacterium rhizogenes, etc.

Active Publication Date: 2011-11-30
北京中植科创生物技术有限公司
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

[0003] The application of Agrobacterium rhizogenes on peanuts is mainly to infect wild Agrobacterium rhizogenes, and to extract resveratrol in peanut roots in the induced root system, and there is no report on the transformation of exogenous genes by Agrobacterium rhizogenes

Method used

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  • Method for establishing agrobacterium rhizogene mediated peanut root inducing system, and application thereof
  • Method for establishing agrobacterium rhizogene mediated peanut root inducing system, and application thereof
  • Method for establishing agrobacterium rhizogene mediated peanut root inducing system, and application thereof

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Embodiment Construction

[0066] LB solid medium, LB liquid medium,

[0067] The co-culture medium is MS basal medium, and the content of acetosyringone is 5 μmol l -1 -100mol l -1 .

[0068] The subculture medium was MS basal medium.

[0069] The above-mentioned medium can be commercially available, or can be prepared by oneself.

[0070] The vector pGFPGUSplus, Agrobacterium rhizogenes K599, is stored in the applicant's laboratory and can be freely distributed to the outside world. pCAMBIA2300, commercially available.

[0071] Peanuts are any commercially available variety.

[0072] 1. Agrobacterium rhizogenes-mediated microinjection infecting peanuts

[0073] 1) Shell the peanuts, place in 75% ethanol for 1 min, discard ethanol, place in 0.1% mercuric chloride for 4 min, wash with sterile water for 3 times, place on sterile paper until dry;

[0074] 2) Separate the two cotyledons of the peanut with a sterile scalpel, place the cotyledon containing the complete embryo on the basic medium, and ...

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Abstract

The invention relates to a method for establishing an agrobacterium rhizogene mediated peanut root inducing system, and application thereof, belonging to the field of biotechnology. According to the invention, the peanut root system is induced by using agrobacterium rhizogene and a microinjection method to obtain a chimeric plant; simultaneously, the method of the system is optimized, i.e. the peanut is infected by performing microinjection on the epicotyl as an explant; the highest conversion efficiency is 61%; and simultaneously, the bioactivities of cry8Ea1 and cry8Ha1 genes optimized by codon on the larvae of holotrichia parallela are verified. The invention successfully establishes a peanut root inducing system, provides a quick verification method for inducing a peanut plant from an extraneous gene, and provides a technical support for further acquiring genetically modified peanut plants.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for establishing a peanut root induction system mediated by Agrobacterium rhizogenes and its application. Background technique [0002] Agrobacterium rhizogenes is a Gram-negative bacterium that causes hairy roots of plants (Moore L., Warren G., and Strobel G. Involvement of a plasma in the hairy root disease of plants caused by Agrobacterium rhizogenes.Plasmid , 1979, 2:617-26.), and synthesized opines. The large plasmid in Agrobacterium rhizogenes determines the type of opine synthesized by infected plants. According to the difference of secreted opine, Agrobacterium rhizogenes can be divided into cucumine, isocucumine, mannopine and agropine type 4 kind. The rol gene in Agrobacterium rhizogenes was integrated into the plant genome and expressed, causing the infected tissue to differentiate into a large number of roots. Due to its high infection efficiency and wide rang...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/06A01H5/00
Inventor 耿丽丽束长龙张杰宋福平黄大昉
Owner 北京中植科创生物技术有限公司
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