60 results about "Molecular fluorescence" patented technology

The invention discloses an indole-benzothiadiazole derivative as well as a preparation method and application thereof. The general structural formula of the indole-benzothiadiazole derivative provided by the invention is shown in the specification, wherein R is selected from one of methyl piperazinyl, diethylamino, azetidinyl and pyrrolidinyl. The derivative can be specifically combined and fluorescently labeled with mitochondrial RNA in cells, and has good selectivity, high fluorescence signal-to-noise ratio and excellent anti-interference capability in a complex biological system. Meanwhile, the indole-benzothiadiazole derivative provided by the invention is simple in synthetic route, easily available in raw materials, good in stability and convenient to store and use, can be specifically combined with RNA and generate strong fluorescence response, is suitable for real-time detection and imaging of in-vitro and in-vivo mitochondrial RNA, and is an efficient mitochondrial RNA micromolecular fluorescent probe.

The application belongs to the field of new application of functional proteins and mutants thereof, and discloses application of sugarcane ScCBS protein and key domain deletion mutants thereof. The nucleotide sequence of the sugarcane ScCBS protein is shown as SEQ ID NO:1, and the amino acid sequence is shown as SEQ ID NO:2. The key domain deletion mutants of the ScCBS protein are ScCBS△motif1, ScCBS△motif3, ScCBS△motif4 and ScCBS△motif9, which respectively lack 28, 50, 49 and 29 amino acids. The application proves that the ScCBS protein can interact with SCSMV P1 and inhibit the RNA silencing suppressor activity by means of yeast two-hybrid, two-molecular fluorescence complementary experiment and agrobacterium transient expression system; the four mutants all lose the interaction and inhibition activity. Therefore, the ScCBS protein and the key domain deletion mutants thereof can be used for cultivating excellent SCSMV-resistant materials, and have wide application prospects in sugarcane disease-resistant molecular breeding.

The invention discloses application of a downstream transcription factor ONAC131 of an SL signal channel in improving the antiviral property of rice, and belongs to the technical field of biology. Experiments such as transcriptome sequencing screening, yeast two-hybridization and bimolecular fluorescence complementation verify that ONAC131 is an SL downstream response factor and can interact with MID1 to jointly regulate and control expression of RDR1 and RDR6 and activate an antiviral RNAi pathway. Overexpression of ONAC131 can enhance the resistance of rice to grass stunt virus (RGSV), and otherwise, deletion of ONAC131 can weaken the antiviral effect of rice. The gene supplements an SL downstream regulation framework, provides a new target for rice antiviral molecular breeding, and assists in breeding of new varieties of high-yield and high-resistance rice.

The invention provides a biomarker detection method which comprises the following steps: carrying out fluorescence labeling on a specific biomarker in a biological sample, and carrying out single-molecule fluorescence imaging on the biomarker to obtain multiple frames of original microscopic fluorescence images; and analyzing and processing the multiple frames of original microscopic fluorescence images so as to extract the super-resolution distribution characteristic information of the specific biomarker on the surface of the biological sample. The method comprises the following steps: processing an original microscopic fluorescence image to obtain positioning table data; rendering positioning points in the positioning table data to generate a super-resolution image; performing convolution operation, opening operation, convolution filtering and region segmentation on the super-resolution image to calculate the area of cells, the number of clusters and the area of a single cluster, determining the density of the clusters and the area ratio of the clusters by using the two, and calculating the closest distance between the clusters by using the center points of the clusters; and the accuracy of a biomarker detection result is greatly improved.

The application discloses a tryptamine detection method based on triple emission MOFs and a smart phone. A rare earth metal organic framework (AP-Eu) is used as a luminescent matrix, and a fluorescent guest molecule fluorescein-5-isothiocyanate (FITC) is post-modified to obtain a ratio fluorescent material with triple emission characteristics. Competitive absorption of ligands and tryptamine, pH response of FITC and hydrogen bonding are used to realize fluorescence signal regulation, and then high-sensitivity and high-selectivity fluorescence recognition of tryptamine is realized. The method is further applied to quantitative detection of tryptamine in beer samples, visualization detection of foam and on-site rapid detection assisted by a mobile phone APP. The application provides a safe and efficient tryptamine detection and fluorescence sensing method, and significantly improves detection accuracy and on-site applicability.

The invention discloses a self-feedback intelligent electrochemical wastewater treatment platform and a construction method thereof. The platform modularly integrates an electrochemical water treatment unit, a terephthalic acid probe molecular fluorescence sensing unit, a signal acquisition data processing unit and an electro-hydraulic linkage online control unit. All the units work cooperatively, and the electrochemical water treatment unit generates strong oxidizing hydroxyl radical mineralized pollutants; the fluorescence sensing and data processing unit is used for converting. OH specificity into a fluorescence signal by utilizing probe molecules and mapping the fluorescence signal into. OH steady-state equivalent concentration in real time; and the online control unit implements a model prediction control strategy based on concentration feedback, dynamically adjusts the current density, the electrode potential and the wastewater flow, and realizes real-time adaptive matching of the interface mass transfer and the. OH generation rate. According to the invention, electrogenerated. OH on-line monitoring and dynamic closed-loop optimization of operation parameters are realized, deep mineralization, low energy consumption and long-term stable operation are realized, and the method is suitable for engineering and intelligent operation of electrochemical advanced oxidation treatment of industrial wastewater.

The invention belongs to the technical field of fluorescent probes, and particularly relates to a single-molecule fluorescent probe based on a phenothiazine structure as well as a preparation method and application of the single-molecule fluorescent probe. The preparation method comprises the following steps: firstly introducing an alkyl chain to a nitrogen atom of a raw material to obtain the phenothiazine structure-based single-molecule fluorescent probe, then carrying out regioselective formylation on the raw material to obtain the phenothiazine structure-based single-molecule fluorescent probe, then treating the raw material by adopting aluminum trichloride to realize O-demethylation, and finally carrying out intramolecular cyclization reaction on the raw material and 1H-benzimidazole-2-acetonitrile to obtain the phenothiazine structure-based single-molecule fluorescent probe. The single-molecule fluorescent probe based on the phenothiazine structure prepared by the invention is compatible with orthogonal chemical reaction activity and maintains controllable electron transfer, so that different chemical events are converted into specific optical signals, and selective and mechanism specific fluorescent response to phosgene, copper ions and hypochlorite is realized.

The invention discloses a method for detecting the conformational change of an APO-Cascade protein. Through mutation of Cas8 protein amino acid N376 and Cas5 protein K169 in a Casade protein, the obtained Cas8 and Cas5 labeled by non-natural amino acids can maintain functions, and at the same time, the mutant type of the Cas5 / 8 protein can be subjected to fixed-point fluorescence labeling; according to the method, the Cas6 is used as a template, the N end of the Cas6 is further modified with biotin, and a formed Casade compound can be fixed on a biotin-PEG functionalized single-molecule slide, so that the conformation state of the Casade protein can be truly reflected by using single-molecule fluorescence resonance energy transfer, an effective means is provided for in-vitro research on the structure-function relationship of a system in CRISPR / Casade, and the method has a wide application prospect. And the method is of great significance in designing the Cascade protein with optimized properties and enabling the Cascade protein to play new functions.

The invention discloses an i-motif DNA (deoxyribonucleic acid) modified gold nano flare, a preparation method of the i-motif DNA modified gold nano flare and application of the i-motif DNA modified gold nano flare to detection of miRNA, and belongs to the technical field of nano biological materials and molecular diagnosis. The fluorescence probe is formed by assembling gold nanoparticles and an i-motif molecular fluorescence probe, the i-motif molecular fluorescent probe contains four sections of sequences rich in C basic groups and a target miRNA recognition chain, sulfydryl is marked at the 5'end, and fluorescent molecules are marked at the 3 'end. The probe is prepared by adopting a freeze thawing method, the labeling density of the probe is high, in a constructed detection system, the miR-21 detection limit is as low as 1.01 fM, and single-base mutants can be accurately distinguished; the technology does not need complex sample pretreatment and can be directly used for detecting cell total RNA or miRNA in a plasma sample, and the detection result is highly consistent with RT-qPCR; the problems of low marking efficiency and insufficient sensitivity in the prior art are effectively solved, and an innovative tool with high sensitivity, high specificity and clinical suitability is provided for tumor-associated miRNA molecular diagnosis.

The present invention provides an integrated platform for exciting a molecular fluorescence signal, a detection chip, and an optical packaging structure. The integrated platform for exciting the molecular fluorescence signal comprises, arranged in sequence: an input optical coupling structure (1), a first filtering structure (2), an optical beam-splitter tree structure (3), a filtering structure array (4), a fluorescence excitation structure array (5), and an optical coupling-out structure array (6). The detection chip is used for detecting molecular fluorescence and comprises, arranged from bottom to top in sequence: a fluorescence detection layer (103'), a fluorescence collection layer (102'), and a fluorescence excitation layer (101'). The fluorescence excitation layer (101') comprises, arranged in sequence: an input optical coupling structure (1011'), an optical beam-splitter tree structure (1013'), a fluorescence excitation structure array (1014'), and an optical coupling-out structure array (1015'). The fluorescence excitation layer is further provided with an optical feedback structure (1012'), wherein an input end of the optical feedback structure (1012') is connected to a first output end of one branch of the optical beam-splitter tree structure (1013'). The optical packaging structure comprises an optical fiber module (2') and the detection chip (1'). The integrated platform in the present invention incorporates on-chip filters, providing an integrated fluorescence excitation solution capable of suppressing broadband background light arising from spontaneous fluorescence of lasers or waveguides, thereby improving a signal-to-noise ratio. The detection chip and the optical fiber module in the optical packaging structure in the present invention employ a passive optical coupling scheme, thereby facilitating stability of fluorescence signals and reducing complexity of external device optical paths.

The invention relates to a molecular fluorography detection method combined with liquid chromatography, and belongs to the technical field of analysis and detection. The invention aims to solve the technical problems that the high sensitivity is difficult to maintain while the miniaturization of the existing liquid chromatogram fluorescence detector is realized, and a flexible and low-cost detection scheme is lacked. According to the technical scheme, the method is characterized in that after being subjected to liquid chromatography separation, a sample is conveyed to a silica gel coating preparation plate through a polyether-ether-ketone tube to form a detection area; exciting fluorescence by adopting a laser with a first narrow-pass optical filter; acquiring a series of fluorescence images by using an image acquisition unit with a second narrow-pass optical filter; and finally, extracting RGB (Red, Green and Blue) numerical values of a fixed pixel region through computer image processing software, and generating a chromatogram of which the fluorescence intensity changes along with time. The method is mainly used for rapid and high-sensitivity fluorescence detection in the fields of biomedical analysis, environmental monitoring, food safety and the like.

The application discloses a new method for quantitatively analyzing interface nucleic acid hybridization reaction and application thereof, and comprises the following steps: S1, constructing a DNA tetrahedron interface; S2, imaging the DNA tetrahedron interface through a total internal reflection fluorescence microscope; S3, realizing real-time monitoring of a hybridization event of the DNA tetrahedron interface by adopting double-color co-localization and analyzing a fluorescence signal; and S4, analyzing hybridization kinetics on the DNA tetrahedron interface by recording changes of the fluorescence signal with time. The application uses the DNA tetrahedron interface for single-molecule fluorescence imaging, can realize real-time monitoring of a single nucleic acid molecule hybridization event, improve detection sensitivity and specificity, accurately analyze kinetic parameters of a hybridization reaction, help realize more accurate performance evaluation of a nucleic acid sensor, has important significance for design and optimization of the nucleic acid sensor, and thus plays a greater role in the fields of clinical diagnosis, environmental monitoring and food safety.

The application discloses a kind of digital single molecule enzyme multi-activity characteristic cross-scale heterogeneity dynamic evaluation methods, comprising: the enzyme-template-primer ternary complex of pre-combination completion is diluted to single molecule level to be loaded into the reaction chamber in microwell array chip to carry out rolling circle amplification, and utilize molecular beacon and the fluorescence signal released by amplification product combination;Real-time monitoring and obtaining the fluorescence signal to generate time-resolved fluorescence intensity curve;The slope parameter of the fluorescence intensity curve is extracted to obtain the catalytic synthesis rate by conversion calculation for evaluating enzyme activity;The catalytic synthesis rate value distribution is fitted using n times Gaussian mixture model to obtain the standard deviation for evaluating population uniformity and the coefficient of variation for evaluating stability.The application can realize single molecule resolution, multi-parameter dynamic correlation and environmental interference processing cross-scale analysis simultaneously, and break through the resolution limit of traditional analysis technology by integrating single molecule fluorescence tracking and population statistics.

The application discloses a KRAS small-molecule fluorescent probe and a preparation method and application thereof, a structural general formula of the KRAS small-molecule fluorescent probe is shown as formula (I), and the fluorescence polarization activity determination method based on the small-molecule fluorescent probe has the advantages of simple test system, less required protein types, low dosage, no need of a complex buffer system, small influence of time and solvent and the like compared with HTRF. The method can be used for screening of KRAS wild type and various mutant inhibitors including but not limited to G12C, G12V, G12D, G12A, G12S and G12R, and has a good application prospect.

This invention discloses a triphenylamine-based D-A type AIE molecule targeting STAT3, its preparation method, and its application in the diagnosis and treatment of colorectal cancer. This molecule uses triphenylamine as a donor and pyridinium as an acceptor, linked by a rigid conjugated trans-vinyl group, exhibiting aggregation-induced emission (AIE) properties. The preparation method employs a Knoevenagel condensation reaction, using 4-(diphenylamino)benzaldehyde and 1,4-dimethylpyridinium iodide as raw materials, and proceeds via a nucleophilic addition-dehydration reaction catalyzed by pyrrolidine, with a yield of 50±3% and a purity ≥98%. This molecule exhibits a fluorescence emission wavelength of 640 nm, demonstrates a significant AIE effect, can penetrate the colorectal cancer cell membrane and specifically aggregate, and shows activity against the IC50 of HCT116 and DLD1 cells. 50 The concentrations were 6.41 μM and 11.46 μM, respectively, which exerted anti-tumor effects by inhibiting STAT3 phosphorylation, enabling integrated diagnosis and treatment of colorectal cancer.

The invention discloses a double-terminal fluorescence labeled polymer as well as a preparation method and application thereof. The structural formula of the double-terminal fluorescence labeled polymer is shown as a formula V. In the formula V, n is 30-300, and x is 1-10. The preparation method comprises the following steps: carrying out ATRP (Atom Transfer Radical Polymerization) active polymerization reaction on a monomer by taking 2-bromo-2-methyl propionate 2-hydroxyethyl ester with hydroxyl at the tail end as an initiator to obtain a compound of which the tail end is terminated by hydroxyl and bromo; a hydroxyl terminal in the compound reacts with a perylene fluorescent dye shown in a formula II, a bromine terminal is subjected to an azidation reaction and then is subjected to a click chemical reaction with an ATTO647N dye, and the double-terminal fluorescence labeled polymer shown in a formula V is obtained. The double-fluorescent-molecule end-capped polymer disclosed by the invention has an FRET effect; the double-fluorescent-molecule-terminated polymer can realize single-molecule fluorescence microscopic imaging, and has important significance on basic researches such as micro-nano structure evolution of characterization and analysis materials and the like.

A method for detecting labile zinc (Zn2+) in a biological material is provided herein, the method including: (a) contacting the biological material with a composition including NapBu-BPEA; and (b) imaging the biological material via molecular fluorescence imaging to detect the labile zinc in the biological material. Also provided herein are methods for morphology-correlated detection of labile zinc localization in a subcellular organelle of a living cell and methods for tracking a change in labile zinc localization in a biological material.

The application discloses a single-molecule immunodetection chip based on height limiting and a detection method. The chip comprises a substrate and a detection flow channel formed in the substrate. The detection flow channel comprises: an oil-phase sampling port and a water-phase sampling port; a cross-shaped droplet generator which is connected with the oil-phase sampling port and the water-phase sampling port and is used for generating droplets; and a flat detection cavity, the height H of the detection cavity is less than the diameter D of the droplets in a free state, so that the droplets generated in the droplet generator are extruded in the up-down direction and are changed from spherical shape to flat pie shape after entering the detection cavity. In the application, the flattened droplets formed by the single-molecule immunodetection chip can lock all single-molecule fluorescent centers in a space with a thickness of only 30 microns, which can completely match the large numerical aperture objective focal depth of a high-performance camera. In the application, physical support and flow resistance guidance are provided by the micro-column array, so that the droplets can almost remain static at the imaging moment, and motion blur is completely eliminated.

The application discloses a trace molecular fluorescence detection device, and particularly relates to the field of trace molecular fluorescence detection, which comprises a PCR detector, a placing groove is formed in the PCR detector, a detection piece is arranged in the placing groove, a horizontal movable groove is formed in the inner wall of the placing groove, a fixing mechanism is arranged in the horizontal movable groove, the fixing mechanism comprises a fixing block movably arranged in the horizontal movable groove, an arc-shaped clamping piece is arranged at one end of the fixing block, the arc-shaped clamping piece is connected with the surface of the detection piece, two groups of telescopic rods are arranged at the other end of the fixing block, and the ends, away from the fixing block, of the two groups of telescopic rods are connected with the inner wall of the horizontal movable groove. The detection piece arranged in the PCR detector is used for detecting the to-be-detected object, the fixing mechanism arranged in the PCR detector is used for fixing the detection piece when the PCR detector is in an open state, so that the to-be-detected object can be placed by an operator, and the detection piece is adjusted by an adjusting mechanism when the PCR detector is used for detection, so that the to-be-detected object can be detected.

The invention discloses a fluorescence detection reagent, a method for doping labeled nucleotide by using the reagent, and a kit or a mixture system. The fluorescence detection reagent contains an enzymatic deoxygenation system and a plurality of reducing agents, and does not contain DABCO. The composition is suitable for application scenarios involving detection of fluorescence signals, in particular for application including detection of single-molecule fluorescence signals.

The application belongs to the field of organic molecule fluorescent probe, and a small molecule fluorescent probe HX is rationally designed, and the integration of diagnosis and treatment is realized by monitoring the related physical properties of tumor microenvironment and the photodynamic therapy of tumor. The tumor microenvironment of solid tumor usually has the characteristics of low pH, high viscosity, low oxygen and the like, and it is of great significance to carry out real-time monitoring and effective inhibition on tumor tissue based on the characteristic parameters of tumor site. Herein, a near-infrared fluorescent probe HX is designed, the probe does not respond to the change of surrounding viscosity in a high pH environment, however, in a low pH microenvironment of tumor, the "Donor-π-Acceptor (D-π-A)" structure thereof is opened, and sensitive viscosity response characteristics are exhibited, so that the dynamic monitoring of the tumor microenvironment is realized. In addition, through structure optimization, it is also confirmed that the probe can be used as a photosensitizer with type I reaction for photodynamic therapy of tumor tissue under hypoxic conditions. With the help of a mouse tumor model in vivo, it is confirmed that the probe HX not only has the ability to identify tumor tissue and normal tissue, but also has excellent tumor inhibition effect and good biocompatibility under hypoxic conditions, thereby providing a new technology for the diagnosis and treatment integration design of tumor diseases.

The invention discloses a supramolecular fluorescent probe and a preparation method and application thereof, preparation raw materials comprise pyrene pyridinium salt derivatives and cucurbit [7] uril, an assembly is formed by electrostatic interaction of the pyrene pyridinium salt derivatives and macrocyclic cucurbit [7] uril, and visual detection is carried out by utilizing fluorescence color change generated by damage of heparin to the assembly. The system can directly identify the color change through naked eyes under the high concentration of heparin, and shows a good linear relationship under the low concentration. The probe has high sensitivity, can provide visual color change at 0.1 mu g / mL, and can be evaluated under the condition of no instrument. The supermolecular fluorescent probe disclosed by the invention is simple in synthesis route, can realize visual recognition of heparin response, has remarkable advantages compared with a traditional fluorescent probe, and has a wide biological application prospect.

The invention discloses a supramolecular fluorescent sensing array and application thereof in preparation of a preparation or a kit for early diagnosis of Alzheimer's disease, the supramolecular fluorescent sensing array comprises a plurality of sensing units, and each sensing unit is composed of a macrocyclic main solution and a fluorescent indicator solution; a macrocyclic main body used in the macrocyclic main body solution is selected from at least one of any macrocyclic main bodies; a fluorescent indicator used in the fluorescent indicator solution is selected from at least one of any fluorescent indicators. The supramolecular fluorescence sensing array can accurately distinguish cerebrospinal fluid samples of alzheimer disease transgenic rats in different disease course stages, and a new method is provided for early-stage accurate diagnosis and disease course monitoring of the alzheimer disease.

The invention discloses a method for simultaneously detecting viscosity and hydrogen peroxide (H2O2) by using a molecular fluorescent probe based on an intramolecular charge transfer (ICT) mechanism. The method comprises the following steps: firstly carrying out hydroformylation on strong electron donor triphenylamine, then introducing a boronic acid pinacol ester group into a strong electron acceptor quinoline structure as a response site of H2O2, and then connecting the donor and the acceptor through freely rotatable carbon-carbon double bonds through aldol condensation reaction to form the molecular probe with a stable ICT structure. H2O2 is subjected to specific oxidation, boronic acid pinacol ester groups are eliminated, intramolecular charge transfer is reduced, emission wavelength blue shift is caused, fluorescence is enhanced, and therefore H2O2 detection is achieved; free rotation of double bonds can be inhibited by increasing the viscosity of the system, the radiative transition number of excited state electrons is increased, red color is enhanced, and therefore the viscosity level is detected. The fluorescent probe has the advantages of high speed, strong specificity and high sensitivity in response to viscosity and H2O2, and rapid qualitative and quantitative analysis on viscosity and H2O2 can be realized by utilizing fluorescence change.

The invention belongs to the technical field of environmental analytical chemistry and detection, discloses a soil and water polychlorinated naphthalene environment rapid detection method, and aims to solve the problems that an existing polychlorinated naphthalene detection method is complex in sample pretreatment, long in detection period and high in cost, and a rapid detection technology is insufficient in sensitivity, weak in anti-interference capability and the like. The method comprises the following steps: constructing a free radical chain reaction kinetics system consisting of a chain initiator, a signal report molecule, a chain transfer and regeneration bifunctional reagent and a pH buffer system, taking polychlorinated naphthalene as a chain terminator, and determining the fluorescence quenching rate change of the signal report molecule after the polychlorinated naphthalene is introduced by measuring the fluorescence quenching rate change of the signal report molecule. The ultrasensitive quantitative detection of polychlorinated naphthalene in soil and water is realized. By adopting the technical scheme, the method has the advantages of high sensitivity and signal amplification effect, no need of complex sample pretreatment, environment friendliness, quick response, excellent anti-interference performance and the like.

This invention discloses a multi-layer three-dimensional single-molecule localization super-resolution imaging method and system based on a zoom lens, including step one: pre-experimental preparation, completing the calibration of the electrically adjustable lens zoom parameters and obtaining the astigmatism calibration curve; step two: fixing the sample, focusing, and activating the anti-drift function and synchronous control program; step three: acquiring multi-layer three-dimensional single-molecule scintillation images using a frame-by-frame scanning mode; and step four: performing three-dimensional localization and stitching reconstruction on all acquired single-molecule fluorescence scintillation images. This invention overcomes the problems of slow scanning speed and mechanical inertia associated with traditional axial scanning relying on the mechanical movement of the microscope objective or stage. Furthermore, due to the rapid zooming and accurate resetting capabilities of ETL, a novel frame-by-frame scanning method can be used, solving the problem in traditional layer-by-layer scanning modes where the focal plane of later scans is irradiated by laser for a longer time than the focal plane of earlier scans, thus minimizing the impact of photobleaching on the imaging quality of different layers.

The invention discloses an i-motif DNA molecular fluorescent probe for detecting miRNA based on a rolling circle amplification technology and a detection method, and belongs to the technical field of molecular diagnosis. A nucleotide sequence of the probe contains four sections of cytosine-rich i-motif quadruplex forming regions and a target specific binding chain, a 5'end modified fluorophore and a 3 'end modified quenching group, and the binding chain can be precisely complementary with an RCA product. The detection method comprises the following two steps: 1) mixing a sample with a padlock probe and an RCA enzyme system, and performing constant-temperature incubation in a buffer solution with the pH value of 7.0-7.5 to realize probe cyclization and rolling circle amplification; and 2) adding an i-motif probe and a buffer solution with the pH value of 5.5-6.5, incubating, detecting a fluorescence signal, and quantifying by combining a standard curve. According to the present invention, the optimal acid-base environment of each link is adapted through the splitting amplification and detection steps, the RCA index amplification and probe dual specificity are combined, the miR-21 / 221 / 222 detection limit is as low as 66 / 78 / 60 aM, and the method can be used for early tumor diagnosis, illness state monitoring and prognosis evaluation, and has important clinical conversion value.

The invention discloses a method for preventing and controlling maize chlorotic mottle virus by using an anti-disease gene ZmCDPK7 and application of the anti-disease gene ZmCDPK7, and belongs to the technical field of plant protection. According to the application, through instantaneous silencing of the ZmCDPK7, an overexpression and knockout line of the ZmCDPK7 is constructed, the resistance reaction of the overexpression and knockout line to maize chlorotic mottle virus infection is analyzed, and it is proved that the ZmCDPK7 positively regulates the immunity of maize to the MCMV. In order to further clarify the molecular mechanism of the ZmCDPK7 for regulating and controlling immunity, interaction between the ZmCDPK7 and a key pathogenic factor P31 of the MCMV is discovered and proved through technologies such as yeast two-hybridization and bimolecular fluorescence complementation. ROS detection proves that the ROS content of the ZmCDPK7 knockout strain is higher than that of a wild type after MCMV infection. In biological and abiotic stress reactions, the resistance of the ZmCDPK7 knockout strain to biological and abiotic stress is proved to be reduced.

The invention relates to the technical field of molecular fluorescence, in particular to a polarity-sensitive fluorescent probe based on ICT and a preparation method and application of the polarity-sensitive fluorescent probe. The preparation method of the polarity sensitive type fluorescent probe comprises the following steps: enabling 4, 7-dibromobenzo [c]-1, 2, 5-thiadiazole to react with [4-[bis (4-methoxyphenyl) amino] phenyl] boric acid to obtain 4-(7-bromo-2, 1, 3-benzothiadiazole-4-yl)-N, N-bis (4-methoxyphenyl) benzylamine, and enabling the 4, 7-dibromobenzo [c]-1, 2, 5-thiadiazole to react with [4-[bis (4-methoxyphenyl) amino] phenyl] boric acid to obtain 4-(7-bromo-2, 1, 3-benzothiadiazole-4-yl)-N, N-bis (4-methoxyphenyl) benzylamine. The preparation method comprises the following steps: enabling 4-(7-bromo-2, 1, 3-benzothiadiazole-4-yl)-N, N-bis (4-methoxyphenyl) benzylamine and (6-nitropyridine-3-yl) boric acid to react, so as to prepare the polarity sensitive type fluorescent probe. The polarity-sensitive fluorescent probe can be used as an effective tool for detecting the polarity change of the lipid droplet, and provides auxiliary diagnosis for diagnosis of some diseases related to the lipid droplet by dynamically monitoring the continuous process of the lipid droplet from generation to maturation to decomposition.