Plant salicylic acid in-situ real-time detection method based on needle electrode

A needle-shaped electrode, real-time detection technology, applied in nanotechnology for sensing, measurement device, material electrochemical variables, etc. Accurate detection results and high sensitivity

Inactive Publication Date: 2021-10-15
NANTONG UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, the detection methods of salicylic acid are mostly carried out by gas chromatography-mass spectrometry or liquid chromatography-mass spectrometry. The main limitation of the above methods is that complex and time-consuming sample pretreatment is required before detection. The reason is that plant tissues and cells need to be frozen and crushed. and sample extraction and purification
Since salicylic acid is a small molecule that is prone to redox reactions, the complex sample processing process will lead to the loss of small molecules of salicylic acid, which will reduce its concentration and make it difficult to further improve the detection limit.
More importantly, the complex sample processing process generally requires the use of a large number of plant samples and takes a long time, it is difficult to truly reproduce the real-time concentration of salicylic acid in situ, and it is difficult for researchers to obtain accurate and timely information, resulting in SA It is difficult to conduct in-depth research on the formation and conduction process in plants, as well as its role in plant disease resistance mechanisms

Method used

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  • Plant salicylic acid in-situ real-time detection method based on needle electrode
  • Plant salicylic acid in-situ real-time detection method based on needle electrode
  • Plant salicylic acid in-situ real-time detection method based on needle electrode

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Effect test

Embodiment 1

[0019] The preparation method of the needle-shaped working electrode, its construction flow chart is as follows figure 1 As shown, it specifically includes the following steps:

[0020] 1. Cut the hard platinum wire (r=0.1mm) to 5cm, put it into a beaker, pour ethanol and distilled water into the beaker in turn for 10min ultrasonic immersion treatment; place the ultrasonically treated platinum wire at room temperature to dry; Take out the dried platinum wire, cut off a 3.5~4cm capillary (r=0.3mm), put the platinum wire into the capillary, one end is 2 mm, the other end is 8~13 mm, fix both ends of the electrode with 3M super glue , dry; wrap one end of 8~13mm with copper conductive tape.

[0021] 2. Put 20 μL of liquid carbon glue with a volume fraction of 50% into a 200 μL centrifuge tube, then take out the needle-shaped electrode with copper conductive tape, put its 2mm end into the 200 μL centrifuge tube for 10 minutes, and then dry it; Put 1mL of multi-walled carbon solu...

Embodiment 2

[0025] The detection process of SA standard substance solution comprises the following steps:

[0026] 1. The preparation method of the standard solution is as follows: first prepare 10mM SA solution, and then use buffer solution to dilute, and the concentration of the prepared SA solution is 0.1, 1, 3, 5, 10, 30μM. Among them, the buffer solution is 0.2M, PBS solution of pH 7, and its preparation method is: sequentially prepare 0.2mol / L K 2 HPO 4 solution and KH 2 PO 4 solution, the K 2 HPO 4 Solution KH 2 PO 4 The solution was mixed and the pH was adjusted to 7.

[0027] 2. Preparation of the standard curve: The current peak values ​​of different concentrations of SA were obtained by using the DPV method. Based on the relationship between the current (μA) and different concentrations of SA (μmol / L), the SA concentration standard curve ( Figure 2B ), the current (Y) and SA concentration (X) are linear in the range of 0.1-30μM, and the regression equation is Y=0.836X+...

Embodiment 3

[0029] The detection of plant salicylic acid comprises the following steps:

[0030] 1. Preparation of tomato bacterial leaf spot ( Pst DC3000) liquid, which was sprayed on the cultivated tomato plants as the experimental group, to directly spray the same volume of MgCl 2 Buffer solution to the tomato plants is the control group, and the experimental group and the control group are 3~4 tomato plants, which are respectively placed in two different transparent storage boxes for moisture cultivation; the cultivated tomato plants are fixed with stainless steel needles (r=0.4mm ) take 3 points on its side stem.

[0031] 2. Insert the constructed three-electrode detection system into the lateral stem of the tomato, and connect it with the electrochemical workstation, and use the DPV method to detect the current value of SA; calculate the concentration of plant salicylic acid according to the relational function of the standard curve .

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Abstract

The invention discloses a plant salicylic acid in-situ real-time detection method based on a needle electrode. According to the detection method, salicylic acid in tomato side stems inoculated with PstDC3000 is subjected to in-situ real-time detection by a three-electrode system formed by the needle electrode; the diameter of the needle-shaped electrode is 0.1 mm, and the needle-shaped working electrode is modified by liquid carbon glue and a multi-walled carbon nanotube. The SA detection needle electrode system constructed by the invention effectively reduces the damage to the plant body in the detection process, can realize the in-situ real-time detection of the plant body SA, provides a new research strategy for deep research on the action mechanism of SA in plant disease resistance and stress resistance, and has important theoretical and practical significance.

Description

technical field [0001] The invention relates to a biological detection method, in particular to an in-situ real-time detection method of plant salicylic acid based on needle electrodes. Background technique [0002] Plant salicylic acid is an important endogenous hormone in plants. Its content in plants is very small, but it has an extremely important regulatory function on plant disease resistance. Salicylic acid (SA) is a small molecular compound with a phenolic structure ubiquitous in plants, and the formation of plant innate immunity and systemic acquired immunity requires the participation of SA. Current studies have shown that the function of SA is determined by its relative concentration in plants. Therefore, determining the concentration distribution and dynamic changes of SA in plants is one of the key issues in understanding how salicylic acid regulates plant disease resistance and stress resistance. [0003] At present, the detection methods of salicylic acid are...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/416G01N27/30B82Y15/00
CPCG01N27/416G01N27/30G01N27/301B82Y15/00
Inventor 孙利军姚登兵张亚莉霍逗逗李道东田一然
Owner NANTONG UNIVERSITY
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