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Method for determining superoxide anion radicals in tape grass leaves

A superoxide anion and free radical technology, applied in the direction of electron paramagnetic resonance analysis, etc., can solve the problems of influence on promotion, lack of specificity of spectral peaks, and difficulty in accurate quantification of free radicals, etc. It is easy to master, simple to operate and practical broad effect

Inactive Publication Date: 2012-01-11
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, it has become a commonly used aquatic plant species in toxicology experiments. The relevant researches focus on the oxidative stress caused by various pollutants, but the direct evidence of free radicals induced by pollutant stress is rarely reported.
Yin Ying et al. (2007) cut the freeze-dried leaves of Erythrina fragrans and used EPR to directly detect the free radical content of the leaves. The obtained EPR spectrum peak was a single peak, but this method failed to use the characteristic capture agent to directly capture the living free radicals. It is difficult to quantify the free radicals measured accurately, and the obtained spectral peaks also lack specificity, which affects its promotion.

Method used

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  • Method for determining superoxide anion radicals in tape grass leaves
  • Method for determining superoxide anion radicals in tape grass leaves
  • Method for determining superoxide anion radicals in tape grass leaves

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Determination of Superoxide Anion Free Radicals in Erythrina fragrans Leaves under Microcystin MC-LR Stress

[0030] The submerged plant Vallisneria natans (Lour.) Hara, which is common in freshwater environment in my country, was selected as the test organism. The plants are cultivated from commercial Erythrina chinensis seeds, and the culture solution adopts improved Hoagland nutrient solution (0.1×) and is prepared with tap water after aeration. The culture temperature is 25°C, the light intensity is 4000lux, and the photoperiod is 12 / 12h. Select well-growing seedlings with a height of about 10 cm and move them into a glass tank containing 5 cm of quartz sand, which contains Hoagland dilution. After domestication for several days, add different concentrations of MC-LR, and the exposure concentration is 0, 0.1, 0.5, 1.0, 5.0, 10.0 μg L -1 , each concentration group had 3 parallels, exposed for 14 days, changed the water every other day and added nutrient s...

Embodiment 2

[0034] Example 2 Determination of superoxide anion free radicals in leaves of Erythria galerica under ammonium nitrogen stress

[0035] The submerged plant Vallisneria natans (Lour.) Hara was selected as the test organism. The germination and cultivation of bitter grass plants are the same as in Example 1, and the plant seedlings with a height of about 10 cm are selected and moved into a glass jar containing 5 cm of quartz sand, containing nitrogen-free Hoagland dilution, and adding different concentrations of ammonium chloride to make NH 4 + -N exposure concentration reaches 0, 0.05, 0.1, 3.0 mg L -1 , each concentration group had 3 parallels, exposed for 14 days, changed the water every day and added ammonium chloride mother solution. After the exposure was completed, cut the tip part of the young leaves of the bitter grass, wash them with pure water, and quickly absorb the residual water on the surface of the leaves with filter paper. Quickly weigh about 0.3 g of plant l...

Embodiment 3

[0038] Example 3 Determination of superoxide anion free radicals in leaves of Erythria galerica under cadmium stress

[0039] The submerged plant Vallisneria natans (Lour.) Hara was selected as the test organism. The germination and cultivation of Erythrina fragrans plants are the same as in Example 1, and the culture solution adopts improved Hoagland nutrient solution (0.1×) and is prepared with tap water after aeration. The culture temperature is 25°C, the light intensity is 4000lux, and the photoperiod is 12 / 12h. Select plant seedlings with a height of about 10 cm that are growing well and transplant them into different Cd 2+ For the glass tank with high concentration sediment, the overlying water is aerated tap water balanced for 10 days (the bottom sediment used in the test was collected from a lake area of ​​Taihu Lake, artificially added with different concentrations of chloride and aged for 60 days); each concentration group had 3 parallels, exposed for 14 days. Afte...

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Abstract

The invention discloses a method for measuring superoxide anion radicals in tape grass leaves, belonging to the measuring field of the measuring of superoxide anion radicals. The method comprises the following steps of: firstly, collecting and cleaning the tip parts of fully exposed small and tender tape grass leaves; carrying out Tiron captureing on the Tiron of radicals in a closed operation box in a closed nitrogen environment; and finally measuring by using an EPR (Electron Paramagnetic Resonance) technology. The invention has the advantages of simpler operation and easy grasping, and canmeasure the superoxide anion radicals of the tape grass leaves in a very short time;, find that and the measured radicals are found to be the superoxide anion radicals according to the superfine structure constant and, the spectrogram shape analysis of an EPR spectrogram and the combination of the remarkable decreases of the radical strength of the leaves pretreated by SOD (Super Oxide Dismutase), and therefore the invention is the most direct, accurate and effective method for measuring the superoxide anion radicals. Superoxide anions are sensitive to pollutants and have a favorable dosage effect relation. The invention is also suitable for other aquatic plant and terrestrial plant species and can be suitable for measuring the radicals of the plant root tissue.

Description

technical field [0001] The invention relates to a method for measuring anion free radicals, in particular to a method for measuring superoxide anion free radicals in leaves of Erytheria galerica. Background technique [0002] Reactive oxygen species (ROS) is a general term for oxygen-containing free radicals with high biological activity produced by exogenous oxidants or intracellular aerobic metabolism, including superoxide anion (O 2 · - ), hydroxyl radical ( OH ), hydroperoxyl (HO 2 - ), hydrogen peroxide (H 2 o 2 ), singlet oxygen ( 1 o 2 )Wait. Photosynthesis, respiration, oxidative phosphorylation, β-oxidation of fatty acids and metabolic processes of many oxidases in plant tissues can generate ROS. More and more studies have confirmed that ROS is a normal part of plant metabolism, and it can be used as a signal molecule to induce the stress response of plants to environmental stress, and under certain conditions (such as the stress of pollutants), it can cause...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N24/10
Inventor 王晓蓉姜锦林宋睿任静华杨柳燕顾雪元
Owner NANJING UNIV
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