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Method for screening key metabolites responding to high-temperature stress of poplar

A technology of high temperature stress and metabolites, applied in the field of screening key metabolites in response to poplar high temperature stress, metabolomics analysis of poplar high temperature stress, can solve the problems of less research at the molecular level, and achieve the effect of potential application value

Inactive Publication Date: 2021-02-05
NANJING AGRICULTURAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies on the response of trees to high temperature stress are mainly concentrated on the level of population, individual, organ and cell, while there are few studies on the molecular level

Method used

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  • Method for screening key metabolites responding to high-temperature stress of poplar
  • Method for screening key metabolites responding to high-temperature stress of poplar
  • Method for screening key metabolites responding to high-temperature stress of poplar

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1 handles poplar under temperature environment, obtains control group and high temperature group poplar leaf sample

[0031] Control group (control, CTRL group): treated at 25°C for 6 hours, high temperature group (heat, H group): ramp up the temperature first, rise from 25°C to 35°C within half an hour, stay for 2 hours, and then rise from 35°C to 45°C, and maintain 45°C for 5 hours. Take the 3rd to 4th leaves under the terminal bud, sampling time point: 5h at 45°C (such as figure 1 shown).

Embodiment 2

[0032] Example 2 Preparation of metabolites from leaf samples of high temperature group and control group

[0033] Get 0.06g leaf tissue sample and add in 2ml EP tube, then add in 0.48ml extraction solvent (methanol: H 2 O(v:v)=3:1) and 24μL ribitol solution (2mg / ml); process the extract with a grinder for 4 minutes, ice-water bath for 5 minutes (repeated twice), and refrigerated centrifugation for 15 minutes (4°C, 13000rpm ); Take 0.35ml supernatant and transfer to 2ml GC / MS glass flask. Dry the extract in a vacuum concentrator; add 80 μL methoxyamine salt reagent (commercially available product) to the dried metabolites, and after slow mixing, incubate at 80°C for 30 minutes; then, quickly add 100 μL BSTFA at 70°C Incubate for 1.5h.

Embodiment 3

[0034] Example 3 Detection of metabolites in high temperature group and control group based on GC / TOF-MS

[0035] GC / TOF-MS analysis was performed on Agilent 7890 gas chromatography-time-of-flight instrument configured with DB-5MS capillary column (5% diphenyl, 95% dimethyl polysiloxane). The specific detection and analysis conditions of GC / TOF-MS are: injection volume: 1 μL, splitless mode; carrier gas: helium; front injection port purge flow rate: 3mL min -1 ;Column flow rate: 1mL min -1 ;Column temperature: 50°C for 1 minute, rising to 310°C at a rate of 10°C per minute, and holding for 8 minutes; Front inlet temperature: 280°C; Transfer line temperature: 270°C; Ion source temperature: 220°C; Ionization voltage : -70eV; scanning mode: 50-500m / z; scanning rate: 20spectra / sec; solvent delay: 460s, and the acquisition mode is full scanning mode.

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Abstract

The invention discloses a method for screening key metabolites responding to high-temperature stress of a poplar. The method comprises the following steps: detecting metabolic change rules of poplar leaves under high-temperature stress through a GC / TOF-MS technology, and screening micromolecular metabolites and metabolic pathways of the poplar responding to high temperature, thereby explaining a regulation and control mechanism of the poplar on high-temperature stress response metabonomics level. 67 differential metabolites are screened out, and differential genes are mainly enriched in valine, leucine and isoleucine biosynthesis under high-temperature stress, galactose metabolism, and a metabolic pathway of glycine, serine and threonine. The invention also provides an important referencebasis for tree high-temperature stress research, and provides a valuable reference for woody ornamental plant high-temperature resistance research.

Description

technical field [0001] The invention belongs to the technical field of metabolomics, and relates to a metabolomics analysis method for poplar high temperature stress, in particular to a method for screening key metabolites responding to poplar high temperature stress. Background technique [0002] Climate change is increasing the frequency of extreme high temperatures around the world. High temperature stress is one of the main environmental stresses, which limits the growth, metabolism and yield of plants. Plants, as sessile organisms, have evolved complex and diverse systems to respond to high temperature. For example, plants use antioxidant enzymes to remove excess reactive oxygen species (ROS) produced by oxidative stress, accumulate some osmotic substances (proline, beet alkali, soluble sugar, etc.) to reduce osmotic stress damage. Studies on the response of trees to high temperature stress mainly focus on the level of population, individual, organ and cell, while the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/72G01N30/04G01N30/06G01N30/86G01N30/88
CPCG01N30/02G01N30/04G01N30/06G01N30/72G01N30/8634G01N30/8693G01N30/8696G01N30/88G01N2030/045G01N2030/062G01N2030/067G01N2030/884G01N2570/00
Inventor 滕年军金飚任世雄沈楠冯婧娴崔佳雯陆维超袁国振蓝令王艺陈子琳吴慧君吴垠何岭
Owner NANJING AGRICULTURAL UNIVERSITY
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