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Method for rapidly detecting transgenic anti-aphid cotton

A genetically modified and fast technology, applied in the direction of measuring devices, material analysis by electromagnetic means, instruments, etc., can solve the problems of long detection period of antibiotic method, toxic chemicals, expensive measuring instruments, etc., and achieve fast detection speed and good repeatability , the effect of high accuracy

Inactive Publication Date: 2012-12-05
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The above method of transgenic insect-resistant cotton has the following disadvantages: (1) Both protein and molecular biology methods require expensive measuring instruments, and protein detection requires kits, and nucleic acid detection requires extraction of cotton DNA or RNA and synthesis of exogenous genes. Specific primers are expensive and can produce toxic chemical substances during the determination process; (2) The antibiotic method has a long detection cycle; (3) The artificial feed method is suitable for transgenic cotton resistant to chewing mouthparts. Detection of Transgenic Cotton Not Suitable for Resistance to Piercing-sucking Aphid

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] ⑴ Preparation of equipment and materials to be tested

[0022] The detection instrument is a 4-channel electrical penetration graph (EPG). The cotton strains to be tested are A1 and A2. The growth period is the seedling stage with 4 true leaves. The 4-channel insect penetration graph detects 4 plants each time. Cotton; connect the back plate of cotton aphid wingless adults with the gold wire electrode with water-soluble conductive silver glue, and place it on a cotton seedling leaf to adapt to it for 1 hour. Only one cotton aphid can be placed on each cotton plant; then the gold wire The other end of the electrode is connected to a channel of the piercing-sucking potentiometer, and each channel is connected to a gold wire electrode;

[0023] ⑵ Detection and recording

[0024] The detection starts, and the recording of the piercing and suction potentiometer starts. The output signal is recorded on the hard disk of the computer through the analog-to-digital conversion ca...

Embodiment 2

[0031] The operating conditions are the same as in Example 1, except that the cotton strains to be tested are B1 and B2 respectively. The calculation results show that the average time of the waveform corresponding to the passive sucking of juice by the cotton aphid mouthpiece on the B1 cotton leaf phloem sieve is 5.54%, which is less than 10%. According to the judgment standard, B1 is a transgenic aphid-resistant Cotton; the average value of the total time corresponding to the waveform of the cotton aphid mouth needle passively sucking plant juice in the phloem sieve of cotton leaves in the cotton leaf phloem sieve is 28.27%, which is greater than 20%. According to the judgment standard, B2 is non-transgenic Aphid-resistant cotton.

[0032] The line B1 to be tested is a transgenic aphid-resistant transgenic cotton, and B2 is its parent non-transgenic conventional cotton, so the experimental results of the present invention are consistent with reality.

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Abstract

The invention discloses a method for rapidly detecting transgenic anti-aphid cotton, wherein the instrument for the detection is an insect piercing and sucking potential instrument. The method comprises the steps of: connecting a cotton aphid with a gold wire electrode; detecting and recording electrical signals generated when the cotton aphid pierces and sucks to feed on cotton to be detected; finishing after recording for a period of time, analyzing and calculating wave forms generated when the cotton aphid pierces and sucks to feed on the cotton to be detected and appearance time of the wave forms; and judging whether the cotton is the transgenic anti-aphid cotton or not: if the percentage of the appearance time summation of the corresponding wave forms generated when cotton aphid scalpella passively sucks juice in a blade phloem sieve tube of the cotton to be detected is less than 10% of the detection time, the cotton strain is the transgenic anti-aphid cotton and if the percentage of the corresponding appearance time summation of the wave forms generated when the cotton aphid scalpella passively sucks juice in the blade phloem sieve tube of the cotton to be detected is more than 20% of detection time, the cotton strain is non-transgenic anti-aphid cotton. Compared with a conventional detection method, the method provided by the invention has the advantages of high detection speed, simpleness, low cost, no pollution, good repeatability and high accuracy.

Description

technical field [0001] The invention belongs to the technical field of safety evaluation of transgenic plants, in particular to a rapid detection method for transgenic aphid-resistant cotton. Background technique [0002] At present, the detection technology of transgenic insect-resistant cotton mainly involves the following: (1) protein detection method: this method is based on the principle of immunology, and is mainly established for the specific detection method of the target protein expressed by exogenous genes in transgenic plants; (2) ) Nucleic acid detection method: mainly use polymerase chain reaction (polymerase chain reaction, PCR) to detect exogenous genes and marker gene sequences; After 3 to 7 days, observe the color change of the leaves. The leaves of transgenic plants grow normally, and the leaves of non-transgenic plants appear green strips; (4) Artificial feed feeding method: Chinese patent 200610064877 discloses a cottonseed that uses transgenic insect-re...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/00
Inventor 汤清波闫凤鸣崔金杰马英雒珺瑜卢绍辉白素芬李为争高佳
Owner HENAN AGRICULTURAL UNIVERSITY