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Method for detecting wheat yellow mosaic virus-resisting transgenic wheat metabolites

A technology of genetically modified wheat and a detection method, which is applied in the field of gas chromatography-mass spectrometry combined detection, can solve problems such as the threat of wheat production, and achieve the effect of obvious signal intensity

Active Publication Date: 2014-05-21
中检科(北京)测试技术有限公司
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Problems solved by technology

At the same time, because the main wheat varieties in the disease-prone areas are sensitive to this disease, it poses a serious threat to wheat production.

Method used

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  • Method for detecting wheat yellow mosaic virus-resisting transgenic wheat metabolites
  • Method for detecting wheat yellow mosaic virus-resisting transgenic wheat metabolites
  • Method for detecting wheat yellow mosaic virus-resisting transgenic wheat metabolites

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0025] Example Detection method of metabolites in wheat yellow mosaic virus-resistant transgenic wheat based on gas chromatography-mass spectrometry

[0026] 1 Experimental materials and methods

[0027] 1.1 Experimental materials

[0028] Wheat yellow mosaic virus-resistant transgenic wheat 72KD and its non-transgenic parent Yang 11 were provided by China Agricultural University.

[0029] 1.2 Experimental reagents

[0030] Acetonitrile, Ispropyl alcohol, pyridine, Methoxyamine hydrochloride, N-methyl-N-trimethylsilyltrifluoroacetamide (MSTFA), myristic acid (Myristic acid).

[0031] 1.3 Experimental Instruments

[0032] Gas chromatography instrument Agilent7890A-8975C, thermal mixer MS-100, mixing ball mill MM-400, vacuum concentrator RVT4104-203, centrifuge D-37520, ultrasonic instrument SB-5200, rice huller JLGJ-45.

[0033] 1.4 Experimental steps

[0034] Dehull the seeds; use a mixing ball mill to grind the seeds into fine powder; divide the powder into 100mg portio...

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Abstract

The invention provides a method for detecting wheat yellow mosaic virus-resisting transgenic wheat metabolites on the basis of gas chromatography-mass spectrometry technology (GC-MS). The wheat yellow mosaic virus-resisting transgenic wheat metabolites are optimized on the basis of the GC-MS detection method, the type and proportion of an extracted solution, extracting time, volume of the extracted solution, deriving process temperature and time are respectively optimized, the relative peak area is at an optimal detection state under the extracting condition of the extracted solution that the ratio of water to acetonitrile to isopropanol is 2:3:3 when 1mL of extracted solution is added for extracting for 20min, and more metabolites components can be detected. Under the deriving condition that the temperature is 30DEG C and the extracting time is 90min, the relative peak area is large, and the signal intensity is obvious. By adopting the detection method, reliable and effective data can be provided for the study of the metabonomics of the wheat yellow mosaic virus-resisting transgene wheat, and a foundation can be laid for the safety evaluation of the transgenic crop.

Description

technical field [0001] The invention relates to gas chromatography-mass spectrometry detection technology, in particular to a detection method for wheat yellow mosaic virus-resistant transgenic wheat metabolites based on gas chromatography-mass spectrometry technology. Background technique [0002] With the maturity of biotechnology and the deepening of genetic engineering, there are more and more varieties of transgenic crops. The safety of genetically modified products has always been the focus of global attention, which is directly related to human health and ecological environment safety. At present, metabolomics method has become one of the important tools for the safety evaluation of genetically modified products. Among them, gas chromatography-mass spectrometry (GC-MS) technology is favored by metabolomics researchers due to its advantages of high resolution, large peak capacity, high sensitivity and short analysis time. As the most valuable analytical tool, in addi...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
Inventor 付伟王玲贺艳朱振营
Owner 中检科(北京)测试技术有限公司
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