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Method for rapid and efficient genetic transformation of brachypodium distachyon by inflorescence dip dyeing

A technology of Brachypodium biscarp and genetic transformation, applied in the field of rapid and efficient genetic transformation of Brachypodium biscarp, can solve problems such as somatic mutation, and achieve the effects of eliminating somatic mutation, good genetic stability and high transformation efficiency.

Inactive Publication Date: 2020-01-10
SHANGHAI CHENSHAN BOTANICAL GARDEN
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AI Technical Summary

Benefits of technology

This patented new technique allows scientists to modify crops without having to grow them from scratch or damaging their roots during conventional methods like crop rotation techniques that involve growing seedlings on soil. It uses an agro-bacillus bacteria called Silverman' s disease virus (Agrobacterum) which causes tiny holes within its DNA when it infects specific plant cells. These small holes allow nutrients such as nitrates to flow through while preventing other substances from entering these areas causing damage. By doing this, researchers could study how different types of genes affect growth conditions and potentially improve agricultural production.

Problems solved by technology

This patents describes methods for transforming brachyphyllum vegetabile called Loxyspiraceae, specifically Grapefruit borer(GFB). These technical problem addressed by these inventors relates to improving agronomic performance through gene modification technology.

Method used

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  • Method for rapid and efficient genetic transformation of brachypodium distachyon by inflorescence dip dyeing
  • Method for rapid and efficient genetic transformation of brachypodium distachyon by inflorescence dip dyeing
  • Method for rapid and efficient genetic transformation of brachypodium distachyon by inflorescence dip dyeing

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Embodiment 1

[0050] 1. Transformation of Agrobacterium and identification of positive clones

[0051] Add the plasmid DNA (pCAMBIA1300, map) containing the hygromycin resistance gene (HYG (R), nucleotide sequence shown in SEQ ID NO.3) to 50 μl of Agrobacterium competent cells (the Agrobacterium strain adopted is EHA105) Such as figure 1 Shown) 0.1-1μg, then ice-bath for 30min; put in liquid nitrogen for 5min, then immediately put in 37℃ water bath for 5min; take out the centrifuge tube, ice-bath for 2-5min, add 0.5ml YEB liquid medium, 28℃, 220rpm Shake the culture for 3-5 hours; take out the bacterial solution and apply it on the YEB solid medium containing the corresponding antibiotics kanamycin and rifampicin (both at a concentration of 50mg / L), and incubate it upside down at 28°C for 2-3 days in an incubator; Take a single colony and do colony PCR identification. The PCR system is: in a 20μL reaction system, Taq DNA polymerase 1.0U, dNTP 0.25μmol / L, primer 0.5μmol / L, 50ng template DNA...

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Abstract

The invention discloses a method for rapid and efficient genetic transformation of brachypodium distachyon by inflorescence dip dyeing. The method comprises the following steps: A, performing transformation of agrobacterium tumefaciens EHA105 and identification of positive clone; b, performing dip-dyeing brachypodium distachyon inflorescences with agrobacterium tumefaciens; and C, obtaining a transgenic brachypodium distachyon T0 generation positive plant. Through an agrobacterium-mediated inflorescence dip-dyeing genetic transformation method, the culture process of calluses is avoided, a large amount of time can be saved, and an effective method for rapidly obtaining the transgenic plants is provided for genetic improvement of crops.

Description

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Claims

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Application Information

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Owner SHANGHAI CHENSHAN BOTANICAL GARDEN
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