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A microelectrode biosensor for in-situ real-time monitoring of the fructose content of a plant and applications thereof

A biosensor and real-time monitoring technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of plant damage and failure to reflect real-time dynamic changes, etc., and achieve the effect of easy mastery and simple operation

Active Publication Date: 2017-03-22
NONGXIN TECH BEIJING CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of these methods are in vitro and static methods, which require in vitro sampling and cause great damage to the plant; and this method can only reflect the static concentration or cumulative effect at a certain time point, and cannot reflect the metabolites in the plant Real-time dynamic changes when the environment changes

Method used

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  • A microelectrode biosensor for in-situ real-time monitoring of the fructose content of a plant and applications thereof
  • A microelectrode biosensor for in-situ real-time monitoring of the fructose content of a plant and applications thereof
  • A microelectrode biosensor for in-situ real-time monitoring of the fructose content of a plant and applications thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The preparation of embodiment 1 working electrode

[0031] (1) The microelectrode array is prepared by microelectromechanical processing technology (MEMS), and is prepared on a silicon wafer substrate. It is about 4-5cm long, including a reference electrode of Ag / AgCl, a platinum counter electrode and a gold working electrode, and the exposed The length of the conductive part is about 10-20mm; the microelectrode is placed in 0.5M dilute sulfuric acid solution for cyclic voltammetry scanning (-0.2 ~ 1.6V) to obtain a typical cyclic voltammetry spectrum to ensure that the electrode surface is clean.

[0032] (2) Drop-coat 1 mM thioglycolic acid (TGA) on the gold working electrode to obtain a TGA / Au electrode.

[0033] (3) Then the ionic liquid [Bmim]PF 6 Drop-coated on the TGA / Au electrode, then gradually drop-coated 40mg / mL fructose dehydrogenase (FDH) and 1mM potassium ferricyanide medium in PBS solution (pH7.4), left at room temperature for 2h, immobilized fructose de...

Embodiment 2

[0035] Embodiment 2 Application of microelectrode biosensor

[0036] (1) prepare the PBS (pH=7.4) solution that concentration is 0.1, 0.5, 1, 5, 10, 50, 100mM fructose respectively, select 0.25V as working potential, use the biosensor that contains the enzyme electrode that embodiment 1 prepares to carry out Chronoamperometry detection was performed to obtain a set of concentration-current relationship curves, and a standard curve y=0.24x+0.04958 (r=0.9826) for fructose was prepared, with a linear range of 0.1-100mM.

[0037] (2) Potted sweet corn was selected as the experimental material, and the sweet corn fruit in the milk ripening stage was used as the detection object. The microelectrodes were randomly inserted into the corn kernels of sweet corn, connected to the electrochemical workstation, and tested at a working voltage of 0.25V by chronoamperometry. The real-time changes of fructose concentration in sweet corn at milk maturity stage were recorded within 3 days.

[0...

Embodiment 3

[0043] 1) The double-layer enzyme electrode (FDH / [Bmim]PF made by Example 1) 6 ) 2 / TGA / Au, under the working potential of 0.25V, continuously detect the standard solution (0.1,0.5,1,5,10,50,100mM) of different concentration fructose, obtain the relational curve of fructose concentration and electric current, with embodiment 2 .

[0044] 2) Insert the microelectrode randomly into the corn kernels of two different varieties of sweet corn E22 and T26 in the milk ripening stage, connect to the electrochemical workstation, and record the milk ripening stage for a certain period of time at a working voltage of 0.25V by chronoamperometry Real-time changes in fructose concentration in sweet corn. The sampling interval of the on-line monitoring sensor prepared by the invention is 0.1 second, which can reflect the dynamic change of fructose in corn kernels in a certain period of time in real time. However, the HPLC method can only sample a few time points, and then detect after a co...

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Abstract

The invention relates to the technical field of microelectrode biological sensing, and particularly discloses a microelectrode biosensor for in-situ real-time monitoring of the fructose content of a plant. According to the microelectrode biosensor, mercaptoacetic acid is assembled onto a work electrode through a drip coating method, then an ionic liquid [Bmim]PF6 is adsorbed to further modify a mixture of fructose dehydrogenase and a potassium ferricyanide medium micromolecule, then the process is repeated to achieve a multilayer modified work electrode of the ionic liquid and the fructose dehydrogenase, the fructose dehydrogenase can catalyze 5-keto-D-fructose generation from D-fructose, and real-time dynamic responses of the enzyme to fructose changes are recorded through chronoamperometry. Through application of the microelectrode biosensor, in-situ real-time monitoring of the fructose content of the plant can be achieved, a tested sample is free of substantial damage, an obtained data result can reflect content changes of fructose in the plant in real time and dynamically, and practical application and operation are simple and convenient and easy to master.

Description

technical field [0001] The invention relates to microelectrode biosensing technology, in particular to a microelectrode biosensor for in-situ real-time monitoring of plant fructose content and application thereof. Background technique [0002] Plants conduct photosynthesis through leaves to form carbohydrates to supply the growth and development needs of plants. Sugars are the main end products of photosynthesis. Studies have shown that sucrose, glucose, and fructose are the main sugars in plants. The synthesis, transportation, and Accumulation is closely related to crop yield and quality, and is involved in the regulation of plant cell osmotic pressure and signal transduction under stress. For example, tomatoes cultivated in facilities in warm seasons often have a sub-high temperature of 30-35°C. If they are kept at this temperature for a long time, the photosynthesis of plant leaves will decrease, the formation of photosynthetic products will decrease, and the metabolism o...

Claims

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Application Information

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IPC IPC(8): G01N27/327G01N27/416
CPCG01N27/327G01N27/416
Inventor 李爱学王成胡叶侯佩臣王晓冬罗斌宋鹏潘大宇路文超周亚男
Owner NONGXIN TECH BEIJING CO LTD
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