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
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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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