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Method for measuring content of nonstructural carbohydrates in rubber tree bark and xylem

A carbohydrate and determination method technology, applied in the direction of color/spectral characteristic measurement, etc., can solve the problems of no determination of xylem starch content, no starch content, long time-consuming, etc., to achieve the effect of short time-consuming, simple steps, and little damage

Inactive Publication Date: 2014-07-09
RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there are few studies on the extraction of tree starch. In my country, Liang Shangpu measured the starch content of rubber trees in the early 1980s (Taping System Reform Papers 1, p148-153). After the bark is directly dyed, it is graded according to the color depth, and the starch grade can be distinguished by naked eyes. This method does not quantify the starch content, nor does it measure the starch content of the xylem
In foreign countries, Canadian researchers used the method of enzymatic hydrolysis to extract and measure the starch of the leaves and roots of poplar and pine trees (Pak S C and Simon M L, 2004). Since this method has many steps in the operation process and takes a long time, The amount of samples required is large. For rubber trees, the bark is the guarantee for tapping rubber. Trying to protect the trees used in the test is also the basis for ensuring that the follow-up test can be carried out smoothly. Therefore, the method is not suitable for the detection of tree starch of rubber trees.
[0004] How to establish a method for extracting and measuring non-structural carbohydrates in rubber tree bark and xylem has become a technical problem to be solved in this field. At present, there are no reports about the technical achievements in this regard.

Method used

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  • Method for measuring content of nonstructural carbohydrates in rubber tree bark and xylem
  • Method for measuring content of nonstructural carbohydrates in rubber tree bark and xylem
  • Method for measuring content of nonstructural carbohydrates in rubber tree bark and xylem

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Experimental program
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Effect test

Embodiment 1

[0033] Establish a standard curve for soluble sugars

[0034] Use 80% ethanol solution to prepare glucose (reducing sugar) and sucrose (non-reducing sugar) solutions with a concentration of 1mg / ml respectively; : 1 volume ratio mixed to form 100μl soluble sugar mixture, 150mg anthrone was dissolved in 100ml dilute sulfuric acid (70ml concentrated sulfuric acid plus 30ml water) to prepare anthrone reagent. Mix sucrose and glucose solution in proportion (the mixing ratio is as shown in the table below), add 3ml of anthrone reagent to 100μl of the mixed solution, keep it warm at 90°C for 15min, and scan the maximum light absorption:

[0035] Table 1 Determination of light absorption peak of soluble sugar

[0036] Sucrose volume (μl) 20 30 50 70 80 Glucose volume (μl) 80 70 50 30 20 Scan the highest absorption peak (nm) 627 626 627 626 626

[0037] It can be seen from the above table that the scanning absorption peak of the light absorption o...

Embodiment 2

[0042] Establish standard curve for starch

[0043] 80%Ca(NO 3 ) 2 Preparation:

[0044] 80gCa(NO 3 ) 2 .2H 2 Dissolve O in water and make up to 100ml.

[0045] I 2 - Preparation of KI solution:

[0046] 1.3gI 2 Add 3.5gKI, and after dissolving, the volumetric flask is set to 100ml.

[0047] The 1mg / ml starch mother liquor was mixed with 80% Ca(NO 3 ) 2 Dilute to 50μg / ml, 100μg / ml, 200μg / ml, 300μg / ml, 400μg / ml starch solution.

[0048] Take 2ml of starch solutions with different concentrations, add 100μl I 2 -KI solution, measure the light absorption at 580nm, draw the standard curve, the curve equation is: y=177.84.00x(r 2 =0.9942) (see figure 2 ).

Embodiment 3

[0050] Obtain bark and xylem soluble sugar extracts and starch extracts from bark and xylem samples, respectively

[0051] a) Sample

[0052] Sampling the rubber tree bark with a small hole puncher with a diameter of 0.5cm, directly poke the hole puncher into the bark by hand, and take out the corresponding bark from the bark as a bark sample; because the rubber tree wood is hard, it cannot be extracted with a sampler Xylem sample, therefore, at the bark sampling port, use a small electric drill to drill a hole in the xylem perforated part to sample wood chips, as a xylem sample, the drilling depth is 3.0cm, and block the bark and xylem sampling port with iron nails of appropriate size after sampling .

[0053] b) Grind the bark and xylem samples with liquid nitrogen respectively, take 10-200mg after oven drying at 40°C, extract with 0.5ml-4ml80% ethanol at 80°C for 40min, mediate and stir in the middle, centrifuge at 12000rpm×10min, collect the supernatant and residue Add 8...

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Abstract

The invention relates to a method for measuring content of nonstructural carbohydrates in rubber tree bark and xylem, and the method is characterized by comprising following steps of respectively establishing a standard curve of light absorptive quantity of soluble sugar on a position of 626nm and a standard curve of light absorptive quantity of starch on a position of 580nm; respectively extracting soluble sugar extraction solution and starch extraction solution in a bark sample and a xylem sample after sampling the rubber tree bark and the rubber tree xylem; adding 3ml anthracenone reagent in 100 micro liters soluble sugar extraction liquid, keeping the temperature of 90 DEG C for 15 minutes, measuring the light absorptive quantity at the position of 626nm after the soluble sugar extraction liquid is cooled to the room temperature, and respectively calculating the content of the soluble sugar in the bark and the xylem referring to the standard curve of the soluble sugar; and adding 100 micro liters I2-KI solution into 2ml starch extraction solution, measuring the light absorptive quantity at the position of 580nm, and respectively calculating the content of starch in the bark and the xylem samples referring to the standard curve of the starch.

Description

Technical field: [0001] The invention relates to a method for measuring the content of non-structural carbohydrates in rubber tree bark and xylem, aiming at the detection of soluble sugar and starch. Background technique: [0002] Natural rubber (cis polyisoprene) is an important raw material for commercial, defense and transportation industries (Ko J H et al., 2003). At present, about 90% of the natural rubber needed in the world comes from Hevea brasiliensis. The mobilization of starch plays a non-negligible role in latex regeneration and compensation for continuous rubber tapping damage. The starch in the xylem can be regarded as a long-term storage source, while the starch in the bark only serves as an immediate buffer. Soluble sugars in the xylem serve as a transport pool, and soluble sugars in the bark can be seen as an intermediate, ready-to-use store. The research on the growth and decline of starch in the bark and xylem is of great significance for us to analyze ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/31
Inventor 魏芳校现周罗世巧吴明仇键杨文凤郑乾坤
Owner RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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