Sample pretreatment method for plant metabolome analysis

A technology for sample pretreatment and metabolomics, which is applied in the field of rapid processing of metabolome analysis, which can solve the problems of low repeatability of omics analysis, and achieve the effect of ensuring homogenization effect, short sample processing time, and improving repeatability.

Active Publication Date: 2016-02-03
TEA RES INST CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The purpose of the present invention is to provide a fast and efficient metabolome analysis and rapid processing technology for plant tissue samples, so as to solve the disadvantages of the prior art and overcome the problems caused by factors such as pollution and uneven sample mixing in the prior art. Omics analysis has low reproducibility and other defects

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  • Sample pretreatment method for plant metabolome analysis
  • Sample pretreatment method for plant metabolome analysis
  • Sample pretreatment method for plant metabolome analysis

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Experimental program
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Effect test

Embodiment 1

[0029] Embodiment 1 tea tree shoot and leaf sample pretreatment

[0030] Two-year-old cutting seedlings of Longjing 43 tea tree were used as materials. The tea tree samples of the control group were cultured at a constant temperature of 15°C, and the samples of the experimental group were cultured at a constant temperature of 25°C. Six biological replicates were set up in the experiment, and samples were taken independently for detection. After 1 week of culture treatment, shoot leaves were taken for metabolomics analysis.

[0031] The patented method is: the leaf sample is immediately put into a 50ml centrifuge tube and inactivated in liquid nitrogen (the centrifuge tube is placed vertically in the container 6 filled with liquid nitrogen in advance, and the liquid level of the liquid nitrogen in the container 6 is lower than that of the centrifuge tube. The nozzle of the main body 1, the same below). Each tube of sample is about 30g (1 / 3 of the volume of the centrifuge tu...

Embodiment 2

[0039] Embodiment 2: be used for tea tree absorption root sample processing

[0040] 1) Take Longjing 43 tea tree two-year-old cutting seedlings as materials, take tea seedlings and tea tree absorption roots as raw materials, immediately put them into a 50ml centrifuge tube and quickly freeze inactivate them in liquid nitrogen (inject liquid nitrogen into the centrifuge tube, and place the number in container 2 / 3 In the stainless steel insulation lunch box for liquid nitrogen, a 1.5mm small hole is drilled in the middle of the centrifuge tube wall for automatic immersion and outflow of liquid nitrogen), and the sample in each tube is about 30g (1 / 3 of the volume of the centrifuge tube after grinding);

[0041] 2) Sample homogenization and grinding: Pre-cool the 5cm grinding rod in liquid nitrogen for 15 minutes, put it into a centrifuge tube, and cover it with a centrifuge tube cover with a small hole (used to prevent the residual liquid nitrogen from being vaporized and dis...

Embodiment 3

[0044] Embodiment 3: be used for tea tree tender stem sample processing

[0045] Take Longjing 43 tea tree two-year-old cutting seedlings as materials, and take tea seedlings with one bud and two three leaves as raw materials. When sampling, immediately put them into a 50ml centrifuge tube and inactivate them in liquid nitrogen (inject liquid nitrogen into the centrifuge tube, and drill the middle part of the tube wall). 1.5mm hole for automatic immersion and outflow of liquid nitrogen), each tube sample is about 30g (1 / 3 of the volume of the centrifuge tube after grinding), put the 5cm grinding rod in liquid nitrogen for 15 minutes and put it into the centrifuge tube . Cover the centrifuge tube cover with a small hole in advance (used to prevent the residual liquid nitrogen from vaporizing and discharging). Carefully hold the cap of the centrifuge tube with gloves and place it on a vortex shaker for 30 seconds of vibration and grinding. After freezing in liquid nitrogen, r...

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Abstract

The invention discloses a sample pretreatment method for plant metabolome analysis. The method comprises the following steps: sampling and inactivating, sample homogenizing and grinding, centrifugal filtration and the like; various tissue parts are separately sampled, are frozen and inactivated quickly by adopting liquid nitrogen, and are homogenized and ground by high-speed oscillation in a liquid nitrogen environment; a mixed sample is extracted by adopting ultrasonic in an assistant manner after being obtained; finally, protein is removed through high-speed centrifugation. According to the sample pretreatment method, the defects of temperature change, easiness in pollution, serious sample waste, relatively poor repeatability, uneven sample mixing, relatively low work efficiency, high operating danger coefficient, high requirements on the proficiency and work experience of operating personnel and the like are overcome. The sample pretreatment method can be well applied to hard tissue sampling, homogenizing, grinding and extracting of woody plant samples of tea trees and the like, and is a rapid, nondestructive and pollution-free sample pretreatment technology for performing a metabonomics analysis test. The sample pretreatment method can also be applied to a quality component analysis, RNA (Ribonucleic Acid) extraction, a proteomics analysis and the like.

Description

technical field [0001] The present invention relates to a high-efficiency metabolome analysis and rapid processing method for fresh plant tissue samples, specifically, it is a rapid and efficient method for metabolomic analysis and testing of fresh woody plant tissues (including leaves, tender stems, and roots) such as tea trees. Non-destructive and pollution-free sample pretreatment technology. The sample processed by this method can also be used for quality component analysis, RNA extraction, proteomics analysis and other detections. Background technique [0002] Metabolomics technology gradually shows strong advantages in species analysis and research on metabolic mechanism. However, metabolomics methods require high sample pretreatment techniques, metabolites are sensitive to environmental changes, and require high processing conditions. Since many metabolic reactions can occur within 1 second, the sample pretreatment process not only requires absolute low temperature ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01L3/14G01N1/28
Inventor 张群峰石元值马立锋伊晓云刘美雅倪康阮建云
Owner TEA RES INST CHINESE ACAD OF AGRI SCI
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