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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: 2009-08-05
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
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Problems solved by technology

[0017] Technical problem The purpose of the present 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 that there is a 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
  • Method for improving agrobacterium-mediated malus plant conversion efficiency by using ultrasonic

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Experimental program
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Effect test

Embodiment Construction

[0034] (1) Determination of the optimal processing parameters

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

[0036] Ultrasonic-assisted Agrobacterium transformation was performed on apple leaves, and the transformed leaves were transferred to the regeneration medium for pre-expression, and the transformed leaves were stained with GUS staining method, and the instantaneous expression rate of exogenous genes was calculated, and the one with the highest instantaneous expression rate was selected. The combination of ultrasonic power and time processing is the best processing combination.

[0037] result

[0038] The leaves soaked in the Agrobacterium suspension with OD600 (optical density at 600nm) of 0.6 were treated with 100w power ultrasonic waves. It was found that appropriate ultrasonic treatment time can significantly increase the instantaneous expression rate of gus gene. It can be seen from Table 1 that with the increase of ultrasonic treat...

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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 "a method for improving the transformation efficiency of Malus plants mediated by Agrobacterium by using ultrasonic waves" belongs to the field of bioengineering breeding, and is specially used to improve the transformation efficiency of exogenous genes in apple genetic engineering. (2) Background technology [0002] 1 The characteristics and principle of ultrasonic waves [0003] The frequency exceeds the range of sound waves that can be heard (50~2×10 4 times / second) sound waves are called ultrasonic waves, whose wavelength is shorter than ordinary electric waves and can propagate inside solids. Ultrasonic waves include two vibration modes, longitudinal wave and transverse wave. The direction of particle vibration is consistent with the propagation direction of the wave. [0004] The principle of ultrasonic generation is that when a voltage is applied to a material with stretching characteristics, the material will vibrate accordi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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