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Combined determination method of chlorophyll content and proline content

A technology of chlorophyll content and determination method, which is applied in the field of joint determination of chlorophyll content and proline content, can solve problems such as environmental protection, cumbersome steps, and unsafety, and achieve the effects of reducing manpower and material resources, saving test materials, and saving manpower

Inactive Publication Date: 2019-08-23
YANGTZE UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This proline content determination method uses the volatile toxic drug toluene, which is unsafe and not environmentally friendly
[0012] Chlorophyll content and proline content are important physiological indicators of plant stress resistance. In the prior art, chlorophyll content and proline content are generally extracted and measured separately, and the steps are cumbersome.

Method used

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  • Combined determination method of chlorophyll content and proline content
  • Combined determination method of chlorophyll content and proline content
  • Combined determination method of chlorophyll content and proline content

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The present invention provides a kind of joint determination method of chlorophyll content and proline content, and concrete operation steps are as follows:

[0029] 1. Extraction of chlorophyll and proline: Cut 0.2g of leaves into pieces, put them into a 10mL centrifuge tube, add 5mL of 95% ethanol, and soak in the dark for 12h to 72h (24h is the best). Gauze filtration, the filtrate is the extract of chlorophyll and proline.

[0030] 2. Determination of chlorophyll content: Dilute 1 mL of the extract N times with 95% ethanol, mix well, and pour it into a cuvette with an optical path of 1 cm. Using 95% ethanol as a blank, the absorbance was measured at wavelengths of 665nm, 649nm and 470nm.

[0031] Chlorophyll a concentration: C a =13.95A 665 -6.8A 649

[0032] Chlorophyll b concentration: C b =24.96A 649 -7.32A 665

[0033] Carotenoid Concentration:

[0034] After obtaining the concentration of the pigment, calculate the content of each pigment in the tis...

experiment example 1

[0087] 1. Both Example 1 and Comparative Example 1 used 4 leaves and 1 heart of Chinese cabbage leaves as materials, and the determination was repeated 3 times. The experimental data were analyzed statistically using SAS 9.1 software, and the t test was used for significant differences.

[0088] Table 2

[0089]

[0090] As can be seen from Table 2, t=0.30090.05(2) ), the chlorophyll content difference measured by embodiment 1 and the original method (comparative example 1) is not significant.

[0091] 2. Both Example 1 and Comparative Example 2 used 4 leaves and 1 heart of Chinese cabbage leaves as materials, and the determination was repeated 3 times. The experimental data were analyzed statistically using SAS 9.1 software, and the t test was used for significant differences.

[0092] table 3

[0093]

[0094] As can be seen from Table 3, t=0.61400.05(2) ), Example 1 and the proline content difference measured by the original method (comparative example 2) are not s...

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Abstract

The invention provides a combined determination method of chlorophyll content and proline content, and the method comprises the following steps of extraction of chlorophyll and proline: chopping and placing the leaves in 95% ethanol for immersion in the dark for 12 to 72 hours, and filtering the mixture by gauze to obtain a filtrate; determination of chlorophyll content: diluting the extract N times with 95% ethanol, uniformly mixing and pouring the diluted extract into a cuvette, and measuring the absorbance at wavelengths of 665nm, 649nm and 470nm by using the 95% ethanol as a blank; and determination of proline content: adding the obtained extract with glacial acetic acid and an acidic ninhydrin solution, heating in a boiling water bath, performing color comparison at a wavelength of 508nm after cooling, detecting the concentration of proline in the measuring solution from a standard curve, and calculating the percentage of the proline content. The method extracts chlorophyll and proline at one time, avoids grinding, greatly reduces manpower and material resources, saves test materials, and has important significance for plant stress resistance research.

Description

technical field [0001] The invention relates to a method for detecting chlorophyll content and proline content, in particular to a joint determination method for chlorophyll content and proline content. Background technique [0002] Chlorophyll widely exists in green plant tissues such as fruits and vegetables, and combines with proteins in plant cells to form chloroplasts. When plant cells die, chlorophyll is free, and free chlorophyll is very unstable and sensitive to light and heat; under acidic conditions, chlorophyll produces green-brown pheophytin, which can be hydrolyzed into bright green chlorophyll in dilute lye acid salts as well as phytol and methanol. There are two kinds of chlorophyll in higher plants: chlorophyll a and b, both of which are easily soluble in ethanol, ether, acetone and chloroform. There are many methods for determining the content of chlorophyll, among which spectrophotometry is the main method: use a spectrophotometer to measure the absorbanc...

Claims

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Application Information

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IPC IPC(8): G01N21/31
CPCG01N21/31
Inventor 朱进彭玉全李文静
Owner YANGTZE UNIVERSITY
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