Method for improving agrobacterium-mediated malus plant conversion efficiency by using ultrasonic

An Agrobacterium-mediated, transformation efficiency technology, applied in the field of transformation efficiency of exogenous genes, can solve problems such as low genetic transformation efficiency, and achieve the effect of improving genetic transformation efficiency and efficiency

Inactive Publication Date: 2007-04-25
NANJING AGRICULTURAL UNIVERSITY
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AI Technical Summary

Problems solved by technology

[0017] Technical problem The purpose of this invention is to provide a transgenic breeding method that uses ultrasonic technology to improve the genetic transformation efficiency of apples in view of the current situation of low genetic transformation efficiency in the genetic engineering breeding of woody plants such as apples

Method used

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  • Method for improving agrobacterium-mediated malus plant conversion efficiency by using ultrasonic
  • Method for improving agrobacterium-mediated malus plant conversion efficiency by using ultrasonic

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Embodiment Construction

[0034] (1) Determination of optimal processing parameters

[0035] 1) Determine the best parameter method for ultrasonic treatment

[0036] Carry out ultrasonic-assisted Agrobacterium transformation on apple leaves, transfer the transformed leaves to regeneration medium for pre-expression, stain the transformed leaves with GUS staining method, calculate the instantaneous expression rate of foreign genes, and select the one with the highest instantaneous expression rate The combination of ultrasonic power and time processing is the best combination of processing.

[0037] result

[0038] Use 100w power ultrasonic to immerse in OD 600 (Optical density at wavelength of 600nm) The leaves in the Agrobacterium suspension of 0.6 were treated, and it was found that proper ultrasonic treatment time can significantly increase the instantaneous expression rate of gus gene. It can be seen from Table 1 that as the ultrasonic treatment time increases, the instantaneous expression rate of the gu...

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Abstract

The invention belongs to the field of biological engineering breeding disclosing a method of increasing the conversion efficiency of Malus plants mediated by Agrobacterium using ultrasonic. Chipping the Malus octa-edged begonia leaves into small pieces, immerging in 75mg.L-1As Agrobacterium suspension with OD600 0.6 for two minutes, treating with 100 w ultrasonic, transferring to regeneration medium for resistance screening, transforming Malus octa-edged begonia leaves to obtain resistant callus and resistant seedling with a conversion rate of 2.2%. Ultrasonic technology is first applicated in the genetic transformation of Malus plants greatly enhancing transgenic efficiency of Malus plants. The application of ultrasonic assisted Agrobacterium-mediated gene transduction for transducting rolC and transforming Malus rootstock octa-edged begonia receives a 2.2% conversion efficiency, which is consumedly higher than ordinary Agrobacterium mediated method with 0.1 ~ 0.8% conversion efficiency.

Description

(1) Technical field [0001] The invention "using a method for improving the transformation efficiency of Agrobacterium-mediated apple plants by using ultrasonic waves" belongs to the field of bioengineering breeding, and is dedicated to improving the transformation efficiency of foreign genes in apple genetic engineering. (2) Background technology [0002] 1 The characteristics and generation principle of ultrasound [0003] The frequency exceeds the audible range of sound waves (50~2×10 4 Times / second) sound waves are called ultrasonic waves, which have a shorter wavelength than ordinary electric waves and can propagate inside solids. Ultrasonic waves include longitudinal waves and transverse waves. The particle vibration direction is the same as the wave propagation direction is called longitudinal wave, and the vertical wave is called transverse wave. They affect objects at the same time and can easily exchange each other. [0004] The principle of generating ultrasonic waves i...

Claims

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Application Information

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IPC IPC(8): C12N15/87C12N5/10A01H4/00C12N15/82
Inventor 渠慎春章镇蔡斌华丛郁高志红杨梦悦
Owner NANJING AGRICULTURAL UNIVERSITY
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