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Method for quantitative determination of regeneration capability of 1,5-ribulose diphosphate

A technique for quantitative determination of ribulose diphosphate, applied in the measurement of color/spectral properties, etc., can solve the problem of no method, seldom pay attention to the important role, etc., and achieve the effect of stable and reliable share, few steps, and simple calculation

Active Publication Date: 2016-11-09
INST OF GEOCHEM CHINESE ACADEMY OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] When the pentose phosphate pathway was often studied in the past, little attention was paid to its important role in the maintenance of plant photosynthesis under stress conditions, and there was no method for quantitative determination of plant ribulose 1,5-bisphosphate regeneration ability

Method used

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  • Method for quantitative determination of regeneration capability of 1,5-ribulose diphosphate
  • Method for quantitative determination of regeneration capability of 1,5-ribulose diphosphate
  • Method for quantitative determination of regeneration capability of 1,5-ribulose diphosphate

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example

[0055] Example of the present invention: it comprises the following steps:

[0056] First, select the plant to be tested, select the second, third and fourth fully expanded leaves, measure its net photosynthetic rate non-destructively, and take the average value of the three leaves to represent the net photosynthetic rate P of the plant N ;

[0057] Second, after that, immediately take one piece of each leaf whose net photosynthetic rate has been measured, cut it into pieces and mix thoroughly, and take 0.1g of it to extract the enzyme solution;

[0058] Third, measure the phosphofructokinase activity EA expressed by the absorbance change value per unit volume and unit time in the above-mentioned extracted enzyme solution pfk ;

[0059] The 4th, measure the enzyme activity EA of the glucose-6-phosphate dehydrogenase represented by the absorbance change value per unit volume and unit time in the above-mentioned extracted enzyme liquid g6pdh ;

[0060] Fifth, the enzyme acti...

Embodiment 1

[0064] First, select the mulberry tree plants in the control environment, and take the second, third and fourth fully expanded leaves each; use the imported LI-6400XT portable photosynthetic measurement system (LI-COR, Lincoln, NE, USA) to measure the leaves of the plants. The net photosynthetic rate of the plant, taking the average value of the three leaves represents the net photosynthetic rate of the plant P N ; The measurement time is fixed at 15:00-16:00 in the afternoon; the standard measurement conditions are set, and the measurement light intensity is 300 μmol m –2 the s –1 PPFD, temperature set to 25°C, CO 2 Concentration set to 400 μmol mol –1 ;

[0065] Second, after that, immediately take one piece of each leaf whose net photosynthetic rate has been measured, cut it into pieces and mix thoroughly, and take 0.1g of it to extract the enzyme solution;

[0066] Third, measure the phosphofructokinase activity EA expressed by the absorbance change value per unit volu...

Embodiment 2

[0072] First, select the mulberry plant in the stress environment 1, and take the second, third and fourth fully expanded leaves each; use the imported LI-6400XT portable photosynthetic measurement system (LI-COR, Lincoln, NE, USA) to measure the The net photosynthetic rate of the leaves, taking the average of the three leaves represents the net photosynthetic rate of the plant P N ; The measurement time is fixed at 15:00-16:00 in the afternoon; the standard measurement conditions are set, and the measurement light intensity is 300 μmol m –2 the s –1 PPFD, temperature set to 25°C, CO 2 Concentration set to 400 μmol mol –1 ;

[0073] Second, after that, immediately take one piece of each leaf whose net photosynthetic rate has been measured, cut it into pieces and mix thoroughly, and take 0.1g of it to extract the enzyme solution;

[0074] Third, measure the phosphofructokinase activity EA expressed by the absorbance change value per unit volume and unit time in the above-me...

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Abstract

The invention discloses a method for quantitative determination of regeneration capability of 1,5-ribulose diphosphate. The method comprises the following steps: selecting the second, the third and the fourth completely spread leaves, performing non-destructive determination on apparent photosynthesis rate, taking a mean value of three leaves to represent the apparent photosynthesis rate PN of the plant; then taking the leave which has measured apparent photosynthesis rate, shearing the leave and fully mixing the leave, taking 0.1 g of the material for enzyme liquid extraction; respectively determining the phosphofructokinase vitality and glucose-6-phosphate dehydrogenase enzyme vitality expressed by the change value at unit volume and unit time in the extracted enzyme liquid, calculating phosphopentose pathway proportion in the glycometabolism according to a formula, and then multiplying by the apparent photosynthesis rate of the plant leave to obtain the regeneration capability of 1,5-ribulose diphosphate. The method can realize rapid quantitative determination of the regeneration capability of 1,5-ribulose diphosphate, and has the advantages of less step, simple calculation, and comparability and reliability of the determination result.

Description

technical field [0001] The invention relates to a method for quantitatively measuring the regenerative ability of ribulose 1,5-diphosphate in plants, which belongs to the field of biological physiology and biochemistry. Background technique [0002] When organisms are under stress, photosynthesis is often affected by the closure of stomata, and the important reason is that the Calvin cycle's ability to regenerate ribulose 1,5-bisphosphate (RuBP) is reduced. At this time, the pentose phosphate pathway can enter the Calvin cycle by generating ribose-5-phosphate, and generate RuBP through phosphorylation, thereby supplementing the Calvin cycle, so that the photosynthesis of plants under stress conditions can be improved to a certain extent. Replenish. [0003] In the study of plant photosynthesis, the photosynthetic rate is an expression used to indicate the strength of photosynthesis, also known as "photosynthetic intensity". The size of the photosynthetic rate can be expres...

Claims

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Application Information

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IPC IPC(8): G01N21/33
CPCG01N21/33
Inventor 吴沿友姚凯饶森张开艳陆叶赵丽华王瑞杭红涛李海涛刘丛强
Owner INST OF GEOCHEM CHINESE ACADEMY OF SCI
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