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A method for quantifying the share of glycolysis and pentose phosphate pathways in plant sugar metabolism

A technology of pentose phosphate pathway and glycolysis pathway, which is applied in the measurement of color/spectral characteristics, etc., can solve problems such as difficulty in reflecting the influence of metabolic pathways, inability to reflect changes in biological glucose metabolism, and splitting metabolic pathways.

Active Publication Date: 2019-02-15
INST OF GEOCHEM CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the past, the changes in plant glucose metabolism under environmental changes were often studied by studying the changes of one metabolic pathway or several metabolic pathways. Influence, can not reflect the overall changes in biological glucose metabolism

Method used

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  • A method for quantifying the share of glycolysis and pentose phosphate pathways in plant sugar metabolism
  • A method for quantifying the share of glycolysis and pentose phosphate pathways in plant sugar metabolism
  • A method for quantifying the share of glycolysis and pentose phosphate pathways in plant sugar metabolism

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example

[0044] Example of the present invention: it comprises the following steps:

[0045] First, select the plant to be tested, and take each of the second, third and fourth fully expanded leaves;

[0046] Second, the above-mentioned leaves are chopped and mixed thoroughly, and 0.1 g of them are taken to extract the enzyme liquid;

[0047] Third, measure the phosphofructokinase activity EA represented by the absorbance change value per unit volume and unit time in the above-mentioned extracted enzyme solution pfk and glucose-6-phosphate dehydrogenase enzyme activity EA g6pdh ;

[0048] Fourth, the enzyme activity of phosphofructokinase EA pfk Glucose-6-phosphate dehydrogenase enzyme activity A g6pdh Substitute into the formula EA ∑ =EA pfk +EA g6pdh In, the total activity EA of the key enzymes of glucose metabolism was obtained ∑ ;

[0049] Fifth, the enzyme activity of phosphofructokinase EA pfk , Glucose-6-phosphate dehydrogenase enzyme activity EA g6pdh and the total a...

Embodiment 1

[0052] First, select the mulberry plant in the control environment, and get the second, third and fourth fully expanded leaves each;

[0053] Second, the above-mentioned leaves are chopped and mixed thoroughly, and 0.1 g of them are taken to extract the enzyme liquid;

[0054] Third, the enzyme activity EA of the rate-limiting enzyme phosphofructokinase (PFK) in the glycolytic pathway was determined in the leaves of the mulberry tree pfk and pentose phosphate pathway rate-limiting enzyme glucose-6-phosphate dehydrogenase (G6PDH) enzyme activity EA g6pdh ;Calculate the total activity EA of key enzymes in glucose metabolism ∑ ;

[0055] Fourth, the EA pfk 、EA g6pdh and EA ∑ into P EMP =EA pfk / EA ∑ ,P PPP=EA g6pdh / EA ∑ Calculate the glycolytic pathway share P in sugar metabolism EMP and the share of the pentose phosphate pathway in glucose metabolism P PPP ;

[0056] Fifth, and finally, the share of the glycolytic pathway in glucose metabolism P EMP and the share...

Embodiment 2

[0058] The first, select the mulberry plant in the control environment, get the second, third and fourth fully unfolded leaves each;

[0059] Second, the above-mentioned leaves are chopped and mixed thoroughly, and 0.1 g of them are taken to extract the enzyme liquid;

[0060] Third, the enzyme activity EA of the rate-limiting enzyme phosphofructokinase (PFK) in the glycolytic pathway was determined in the leaves of the mulberry tree pfk and pentose phosphate pathway rate-limiting enzyme glucose-6-phosphate dehydrogenase (G6PDH) enzyme activity EA g6pdh ;Calculate the total activity EA of key enzymes in glucose metabolism ∑ ;

[0061] Fourth, the EA pfk 、EA g6pdh and EA ∑ into P EMP =EA pfk / EA ∑ ,P PPP =EA g6pdh / EA ∑ Calculate the glycolytic pathway share P in sugar metabolism EMP and the share of the pentose phosphate pathway in glucose metabolism P PPP ;

[0062] Fifth, and finally, the share of the glycolytic pathway in glucose metabolism P EMP and the shar...

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Abstract

The invention discloses a method for quantitatively determining shares of a glycolysis pathway and shares of a pentose phosphate pathway in plant glucose metabolism. The method comprises the following steps: a to-be-detected plant is selected, a second fully expanded leaf, a third fully expanded leaf and a fourth fully expanded leaf are taken, cut into pieces and mixed sufficiently, 0.1 g of a mixture is taken for enzyme solution extraction, the activity of PFK (phosphofructokinase) and the enzyme activity of G6PDH (glucose-6-phosphate dehydrogenase), which are represented by absorbance change values per unit volume in unit time, in the extracted enzyme solution are determined respectively, and the shares of the glycolysis pathway, the shares of the pentose phosphate pathway as well as the proportion of the glycolysis pathway to the pentose phosphate pathway during glucose metabolism are calculated according to a formula. The method can quantitatively determine the shares of the glycolysis pathway, the shares of the pentose phosphate pathway as well as the proportion of the glycolysis pathway to the pentose phosphate pathway in plant glucose metabolism in different environments, comprises a few steps and is simple in calculation, and a determination result has comparability and reliability and can wholly reflect the disproportionation condition of glucose metabolism of plants when the environment changes.

Description

technical field [0001] The invention relates to a method for quantitatively measuring the share of glycolysis and pentose phosphate pathways in plant sugar metabolism, belonging to the field of biological physiology and biochemistry. Background technique [0002] Under unfavorable habitat conditions, organisms often adapt to changes in the environment by changing their morphology and physiology to gain a chance of survival. The growth and development of organisms are based on metabolism. Therefore, judging the complex changes in the metabolic system of organisms under adverse conditions is the basis for studying the adaptability of plants to the environment. Glucose metabolism is the most important metabolic process to maintain the basic activities of living organisms, providing energy, reducing power and substrates for the synthesis of other biological macromolecules for living organisms. There are many metabolic pathways of glucose in the living body, namely glycolysis p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/33
CPCG01N21/33
Inventor 吴沿友姚凯杭红涛王瑞张开艳饶森陆叶赵丽华李海涛刘丛强
Owner INST OF GEOCHEM CHINESE ACADEMY OF SCI