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A method for simultaneous detection of seven endogenous hormones in wheat leaves

An endogenous hormone, wheat technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of insufficient sensitivity and selectivity, lengthy sample purification procedures, and high use and maintenance costs, achieve objective and easy results, and reduce blind supplementation. The phenomenon of exogenous hormones, the effect of reliable results

Active Publication Date: 2019-07-30
SHANDONG AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0007] The determination of early plant hormones basically used biological testing methods, but due to insufficient sensitivity and selectivity, it has been gradually eliminated
Gas chromatography-mass spectrometry (GC-MS) and monoclonal antibody-based immunoassay are both new technologies for the detection of modern plant hormones. In the GC-MS method, the selective ion detection technology is more rapid and effective. It not only has high sensitivity, and can accurately identify the molecular structure of hormones, but these methods require lengthy sample purification procedures, expensive equipment, high use and maintenance costs
At present, ultra-high performance liquid chromatography tandem mass spectrometry has been reported to analyze the content of plant endogenous hormones. However, because ABA is sensitive to light, JA and SA are unstable when exposed to heat, and the contents of ZR and ZT are extremely low in leaves, especially senescent leaves. GA 3 、GA 4 The difference is only in the number and position of double bonds, carboxyl groups, so this method has not been used to simultaneously detect the seven endogenous hormones ZR, ZT, JA, SA, ABA, GA in wheat leaves 3 、GA 4 report

Method used

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  • A method for simultaneous detection of seven endogenous hormones in wheat leaves
  • A method for simultaneous detection of seven endogenous hormones in wheat leaves
  • A method for simultaneous detection of seven endogenous hormones in wheat leaves

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Embodiment 1

[0056] 1. The hormone is unstable when heated, so it is necessary to pre-cool the mortar for grinding the sample and the spoon for weighing the sample in liquid nitrogen. The extraction agent is prepared in advance and placed in a refrigerator at 4°C for pre-cooling. The test tube rack for the 10mL centrifuge tube Serve in an insulated foam box filled with crushed ice.

[0057] 2. Take the wheat leaves or fresh leaf samples stored in the refrigerator at -80°C, place the samples in an insulated bucket filled with liquid nitrogen, cut off the uppermost and lowermost leaves of the leaves with scissors, leave the middle of the leaves, and put Chop the middle leaves and mix well. Put the mixed sample in a pre-cooled mortar, add liquid nitrogen, grind the sample into powder, accurately weigh 1.0g of the sample powder, add it to a 10mL centrifuge tube, and place the centrifuge tube on the test tube rack.

[0058] 3. Accurately add 4 mL of extractant with a pipette gun (extractant is...

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Abstract

The invention discloses a method for simultaneous detection of seven endogenous hormones in wheat leaves. The endogenous hormones are ZR, ZT, JA, SA, ABA, GA3 and GA4. UPLC-MS / MS is employed in the invention, and chromatographic conditions are that 0.1% acetonitrile is used as a mobile phase A; an aqueous solution of 0.05% formic acid is used as a mobile phase B; and methanol is employed for gradient elution. The method of the invention has the advantages of high specificity, high sensitivity, high throughput and objective and easy-to-analyze results, can determine the imbalance of endogenous hormones during the growth and development of wheat and provide guidance for supplementation of exogenous hormones of wheat, and reduces occurrence of blind supplementation of exogenous hormones.

Description

technical field [0001] The invention relates to a method for detecting endogenous hormones in wheat leaves, in particular to a method for simultaneously detecting seven endogenous hormones in wheat leaves. Background technique [0002] Phytohormones are a type of organic substances produced by the plant's own metabolism, and move from the production site to the action site, and have obvious physiological effects at extremely low concentrations. [0003] The expression, growth and development of plant genes and the response of plants to certain environmental stimuli are all regulated by various hormones in their bodies. Phytohormones are naturally occurring organic compounds in plants. Although the content is very low, they regulate the entire process of plant life activities. Therefore, they are closely related to the basic laws of plant growth and development and the regulation of metabolism. The study of plant endogenous hormones is one of the important contents in the fi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/062
Inventor 李勇骆永丽王振林李文倩常永兰张鑫王元元
Owner SHANDONG AGRICULTURAL UNIVERSITY
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