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Microelectrode biosensor for online detection of IAA (auxin) in living plant and application of microelectrode biosensor

A biosensor and plant auxin technology, applied in the field of microelectrode biosensing, can solve the problems of high time-consuming cost, cumbersome operation, and inability to reflect the real-time dynamic changes of IAA content, and achieve the effect of easy mastery and simple application and operation

Inactive Publication Date: 2017-02-15
BEIJING RES CENT FOR INFORMATION TECH & AGRI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional detection method requires pretreatment such as separation and purification of sample sampling, which is not only cumbersome to operate, time-consuming and costly, but also more destructive to the sample, and can only be tested in vitro, which cannot reflect the real-time dynamic changes of IAA content in the sample

Method used

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  • Microelectrode biosensor for online detection of IAA (auxin) in living plant and application of microelectrode biosensor
  • Microelectrode biosensor for online detection of IAA (auxin) in living plant and application of microelectrode biosensor

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Effect test

Embodiment 1

[0046] The preparation of embodiment 1 working electrode

[0047] 1) Gold nanoparticles were prepared by sodium citrate reduction method. The pH of the gold nanoparticle solution was adjusted to pH 7.6, 100 μl of anti-IAA antibody (1 mg / ml) was added to 400 μL of the gold nanoparticle solution, shaken for 5 minutes, and incubated at 4° C. for 12 hours. The mixture was then centrifuged at 12000 rpm for 30 min at 4°C to remove unattached antibody. Afterwards, the precipitate was dispersed in 1 ml of PBS buffer containing 1% BSA, pH 7.4, to form AuNPs-anti-IAA complexes.

[0048] 2) After cleaning the microelectrodes, immerse in 1M KNO 3 3mM HAuCl 4 In solution, AuNPs-modified microelectrodes were obtained by electrochemical deposition at -0.4V for 200s on the gold working electrode. 2 mM mercaptoundecanoic acid (MUA) was drop-coated on the modified working electrode, and after incubation for 1 h, the excess unreacted MUA was washed away with water, so that MUA could be assem...

Embodiment 2

[0050] Embodiment 2 Application of microelectrode biosensor

[0051] 1) The prepared microelectrode was connected to an electrochemical workstation, reacted with a standard solution of IAA, and the reaction voltage was determined to be 0.2V by electrochemical cyclic voltammetry. The microelectrodes reacted with standard solutions of different concentrations of IAA, and were continuously detected by chronoamperometry at a working voltage of 0.2V. The peak current value was plotted against the concentration of IAA to prepare a standard curve.

[0052] 2) Soybean seedlings cultivated to the 7th day were selected as experimental materials, and the microelectrodes were inserted into the tender stems of the bean seedlings and connected to the electrochemical workstation, and the concentration changes of IAA within 4 days were continuously measured online.

[0053] 3) The tender stems of soybean seedlings cultivated to the 7th, 8th, 9th, and 10th days were respectively taken, and th...

Embodiment 3

[0059] 1) After cleaning the gold working electrode, immerse it in a solution containing 0.5M KNO 3 1mM HAuCl 4 In solution, AuNPs-modified microelectrodes were obtained by electrochemical deposition at -0.4V for 200s on the gold working electrode. Mercaptoundecanoic acid (MUA) was drip-coated on it, and after incubation for 1 h, the excess unreacted MUA was washed away with water, so that MUA could be assembled on the modified gold electrode through the Au-S bond.

[0060] 2) Drop-coat a mixed solution of 50 mM N-hydroxysuccinimide (NHS) and 200 mM 1-3-ethylcarbodiimide hydrochloride (EDC) on the working electrode, and after incubation for 1 h, the AuNPs-anti-IAA complex Drop onto the surface of the modified gold electrode, incubate for 1h to obtain the microelectrode modified by IAA antibody, and further drop-coat 5mM K on the working electrode 3 Fe(CN) 6 Medium body.

[0061] 3) The modified microelectrode is connected to the electrochemical workstation, then the microe...

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Abstract

The invention relates to a microelectrode biosensing technology, and particularly discloses a microelectrode biosensor for online detection of IAA (auxin) in a living plant. The microelectrode biosensor is characterized in that the specific recognition on the IAA is performed through an IAA-modified antibody on a gold (or platinum) working electrode, and the highly sensitive detection of the IAA is performed through AuNPs (gold nanoparticles) and modified IAA-AuNPs compounds deposited on the electrode. The microelectrode biosensor disclosed by the invention has the advantages that dynamic change information of the IAA in a plant body is mastered in situ and in real time by means of continuously monitoring the IAA in the living plant online, the metabolic process of the IAA is understood, and a theoretical basis is provided for understanding the participation of the IAA in the regulation mechanism of a plant living system; the IAA in the living plant can be continuously monitored online by utilization of the microelectrode biosensor disclosed by the invention, and a to-be-detected specimen is prevented from being essentially damaged; an obtained data result can dynamically reflect the IAA content change in the plant body in real time, and the actual application operation is simple and convenient and is easy to master.

Description

technical field [0001] The invention relates to microelectrode biosensing technology, in particular to a microelectrode biosensor for online detection of plant auxin in a living body. Background technique [0002] As a small molecular organic substance in plants, auxin (IAA) is mainly involved in the growth and development of plants. The induction of auxin on plant coleoptile tips, young buds, leaves, etc. usually has a positive and negative duality, mainly depending on its concentration and the physiological status of the plant. Normally, too much auxin will promote the production of ethylene in the plant, disrupt the physiological balance inside the plant, and thus inhibit the growth of the plant. On the other hand, the development of plant fruit is inseparable from auxin. During the development of seeds, the auxin produced can change the distribution of nutrients, so that the ovary develops into fruit. [0003] At present, the commonly used auxin detection methods main...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/30G01N27/26
CPCG01N27/30G01N27/26
Inventor 王晓冬李爱学李璇王成胡叶侯佩臣潘大宇路文超何璐璐张晗
Owner BEIJING RES CENT FOR INFORMATION TECH & AGRI
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