Application of ferroptosis inhibitor 1 in preparation of tunicamycin phytotoxicity inhibitor and pesticide composition

Abstract

The invention relates to the technical field of plant protection, in particular to application of a ferroptosis inhibitor 1 in preparation of a tunicamycin phytotoxicity inhibitor and a pesticide composition. Researches show that the ferroptosis inhibitor 1 can significantly improve the tolerance of plants to the tunicamycin poison, has the effect of relieving the plant poison caused by the tunicamycin, and helps the plants and seeds thereof to normally germinate and grow under the condition of relatively high concentration of the tunicamycin.

Description

Technical Field

[0001] The present application relates to the technical field of plant protection, and in particular to the application of a ferroptosis inhibitor-1 in the preparation of a tunicamycin phytotoxicity inhibitor and a pesticide composition. Background Art

[0002] Tunicamycin (TM) is a nucleotide antibiotic produced by soil actinomycetes. It can be used as an inhibitor of N-acetylglucosamine phosphotransferase (GPT) to inhibit the protein N-glycosylation process in the endoplasmic reticulum (ER) of cells, leading to the accumulation of unfolded proteins in the ER cavity, causing ER stress and cell death. Tunicamycin can inhibit Gram-positive bacteria, yeast, fungi and viruses, and has anti-cancer activity, which has important research and application value.

[0003] The toxic effect of tunicamycin on animal cells can be alleviated by chemical chaperones. The mechanism of action is that chemical chaperones can relieve endoplasmic reticulum stress by helping unfolded proteins to form the correct conformation, reduce unfolded protein response, and alleviate tunicamycin-induced cell death. However, for plant cells, no effective tunicamycin toxicity inhibitors have been found. In order to further study the molecular mechanism of tunicamycin-induced cell death, it is necessary to study tunicamycin plant toxicity inhibitors, which will help to analyze the relevant mechanisms and promote the precise targeted application of tunicamycin.

[0004] Ferrostatin-1 (Fer-1) is a synthetic antioxidant known as a ferroptosis inhibitor that can inhibit Erastin-induced ferroptosis in HT-1080 cells. It may prevent damage to membrane lipids through a reduction mechanism, thereby inhibiting cell death. So far, no reports have been found on the inhibition of tunicamycin activity by ferrostatin-1. Summary of the invention

[0005] Based on this, one or more embodiments of the present application provide an application of a ferroptosis inhibitor-1 in the preparation of a tunicamycin plant toxicity inhibitor and an application in a pesticide composition. The present application research found that the ferroptosis inhibitor-1 has the effect of alleviating the toxicity of plants to tunicamycin and can be used for soil detoxification in agricultural production.

[0006] The technical solution of this application includes the following contents:

[0007] A use of a ferroptosis inhibitor-1 in the preparation of a tunicamycin phytotoxicity inhibitor, wherein the structure of the ferroptosis inhibitor-1 is shown in formula (I):

[0008]

[0009] In some embodiments, the tunicamycin phytotoxicity inhibitor can be used to alleviate the toxic effects of tunicamycin on plants.

[0010] In some embodiments, the toxic effect of tunicamycin on plants includes at least one of inhibiting seed germination and inhibiting seedling growth.

[0011] In some embodiments, the plant includes at least one of a cruciferous plant, a gramineous plant, a leguminous plant, a solanaceous plant, a cucurbitaceous plant, a rosaceous plant, a rutaceous plant, and an umbelliferous plant.

[0012] A method for alleviating tunicamycin toxicity in plants, comprising:

[0013] treating seeds or seedlings of the plant with ferroptosis inhibitor-1;

[0014] The structure of the ferroptosis inhibitor-1 is shown in formula (I):

[0015]

[0016] A use of a ferroptosis inhibitor-1 in the preparation of a pesticide composition, the pesticide composition comprising the ferroptosis inhibitor-1 and a pesticide adjuvant;

[0017] The structure of the ferroptosis inhibitor-1 is shown in formula (I):

[0018]

[0019] In some embodiments, the pesticide composition can be used to alleviate the toxic effects of tunicamycin on plants.

[0020] In some embodiments, the toxic effect of tunicamycin on plants includes at least one of inhibiting seed germination and inhibiting seedling growth.

[0021] In some embodiments, the plant comprises Arabidopsis thaliana.

[0022] A soil detoxification method comprising:

[0023] applying ferroptosis inhibitor-1 to the soil and stirring;

[0024] The structure of ferroptosis inhibitor-1 is shown in formula (I):

[0025]

[0026] The present application provides an application of a ferroptosis inhibitor-1 in the preparation of a tunicamycin plant toxicity inhibitor and a pesticide composition, as well as a method for alleviating tunicamycin toxicity in plants and a soil attenuation method.

[0027] The ferroptosis inhibitor-1 of the embodiment of the present application can significantly improve the tolerance of plants to tunicamycin toxicity, and help plants and their seeds to germinate and grow normally under conditions of higher concentrations of tunicamycin. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] In order to more clearly illustrate the specific implementation methods of the present application or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0029] Figure 1 This is a comparison chart of the detoxification effects of 4-phenylbutyric acid, taurodeoxycholic acid, ferroptosis inhibitor CPX and ferroptosis inhibitor-1 on the Arabidopsis bzip28 / 60 double mutant poisoned by tunicamycin in Example 1 of the present application.

[0030] Figure 2 This is a comparison diagram of the detoxification effects of different concentrations of ferroptosis inhibitor-1 on wild-type Arabidopsis thaliana poisoned by tunicamycin in Example 1 of the present application; Figure 2 A in the middle is a photo comparison of the growth status of wild-type Arabidopsis under different treatment conditions; Figure 2 Middle B is a bar chart comparing the relative weights of wild-type Arabidopsis under different treatment conditions.

[0031] Figure 3 This is a comparison chart of the detoxification effects of different concentrations of ferroptosis inhibitor-1 on tunicamycin-poisoned Arabidopsis wild type, Arabidopsis bzip28 / 60 double mutant and Arabidopsis nrt1.7-2 mutant in Example 1 of the present application.

[0032] Figure 4 This is a comparison result of UPR gene expression in Arabidopsis wild type poisoned by tunicamycin with and without the addition of ferroptosis inhibitor-1 for detoxification in Example 1 of the present application.

[0033] Figure 5 This is a comparative diagram of the detoxification effects of different concentrations of ferroptosis inhibitor-1 on Arabidopsis wild type and Arabidopsis bzip28 / 60 double mutant poisoned by dithiothreitol (DTT) and proline analog L-azaaziridine-2-carboxylic acid (AZC) in Example 1 of the present application.

[0034] Figure 6 This is a comparison chart of the detoxification effect of the ferroptosis inhibitor-1 of Comparative Example 1 of the present application on wild-type and tunicamycin-hypersensitive yeast cells poisoned by tunicamycin; Figure 6A in the middle shows the detoxification effect of ferroptosis inhibitor-1 on wild-type yeast cells poisoned by tunicamycin; Figure 6 Middle B shows the detoxification effect of ferroptosis inhibitor-1 on tunicamycin-hypersensitive yeast cells. DETAILED DESCRIPTION

[0035] The present application will be further described below in conjunction with the embodiments and examples. It should be understood that these examples are only used to illustrate the present application and are not intended to limit the scope of the present application. In addition, it should be understood that after reading the content taught in the present application, those skilled in the art may make various changes or modifications to the present application, and these equivalent forms also fall within the protection scope of the claims attached to the present application.

[0036] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this application belongs. The terms used herein in the specification of this application are only for the purpose of describing specific embodiments and are not intended to limit this application.

[0037] the term

[0038] Unless otherwise specified or incompatible herewith, the terms and phrases used herein shall have the following meanings:

[0039] The term "and / or" used in this article includes any one of two or more related listed items, and also includes any and all combinations of related listed items, and the arbitrary and all combinations include any two related listed items, any more related listed items, or all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", it should be understood that in this application, the technical solution undoubtedly includes technical solutions that are all connected by "logical and", and undoubtedly includes technical solutions that are all connected by "logical or". For example, "A and / or B" includes three parallel solutions of A, B and A+B.

[0040] In the present application, "further", "further", "particularly" and the like are used for descriptive purposes to indicate differences in content, but should not be construed as limiting the scope of protection of the present application.

[0041] In this application, the terms "first", "second", "third", "fourth", etc. are used for descriptive purposes only and cannot be understood as indicating or implying relative importance or quantity, nor can they be understood as implicitly indicating the importance or quantity of the indicated technical features. Moreover, "first", "second", "third", "fourth", etc. only serve the purpose of non-exhaustive enumeration and description, and it should be understood that they do not constitute a closed limitation on quantity.

[0042] In the present application, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0043] In the present application, when it comes to numerical intervals (i.e., numerical ranges), unless otherwise specified, the optional numerical distribution is considered continuous within the above numerical interval, and includes the two numerical endpoints (i.e., the minimum and maximum values) of the numerical range, and each numerical value between the two numerical endpoints. Unless otherwise specified, when the numerical interval only refers to integers within the numerical interval, it includes the two endpoint integers of the numerical range, and each integer between the two endpoints. In addition, when multiple ranges are provided to describe features or characteristics, these ranges can be combined. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all subranges included therein.

[0044] Ferrostatin-1 is a synthetic antioxidant known to be a potent and selective ferroptosis inhibitor that can inhibit Erastin-induced ferroptosis in HT-1080 cells. It may prevent damage to membrane lipids through a reduction mechanism, thereby inhibiting cell death.

[0045] The ferroptosis inhibitor CPX refers to ciclopirox, which is an iron ion chelator.

[0046] Ferroptosis is an iron-dependent, reactive oxygen species (ROS)-regulated programmed cell death characterized by the accumulation of lipid peroxides in cells. The ferroptosis process involves the participation of iron ions, which promote the production of lipid peroxides through the Fenton reaction.

[0047] Tunicamycin is an antibiotic mainly derived from Streptomyces. The main function of tunicamycin is to inhibit the N-glycosylation process. It inhibits N-acetylglucosamine-1-phosphotransferase, preventing the initial step of N-glycosylation, thereby interfering with the synthesis of intracellular glycoproteins and glycolipids, and affecting many physiological functions of cells, such as the secretory function of cells and the function of cell membrane proteins.

[0048] N-acetylglucosamine phosphotransferase (GPT) is a key enzyme involved in the N-glycosylation process. It catalyzes the transfer of N-acetylglucosamine-1-phosphate from UDP-N-acetylglucosamine to dolichol phosphate in the endoplasmic reticulum (ER), which is an important reaction in the initial step of N-glycosylation.

[0049] The endoplasmic reticulum (ER) is an important organelle in the cell. It is a continuous membrane system with tubular, vesicular or flat cystic structures surrounded by membranes. It is divided into the rough endoplasmic reticulum (RER) and the smooth endoplasmic reticulum (SER). Ribosomes are attached to the surface of the rough endoplasmic reticulum, and its main function is to synthesize, process and transport proteins. After the newly synthesized proteins enter the endoplasmic reticulum cavity, they will be modified by folding, glycosylation, etc. The smooth endoplasmic reticulum is mainly involved in lipid synthesis, glycogen metabolism and detoxification.

[0050] N-glycosylation is a post-translational modification of proteins, which is the process of adding oligosaccharide chains to asparagine (Asn) residues of proteins. This process occurs in the endoplasmic reticulum and Golgi apparatus.

[0051] Endoplasmic reticulum stress (ER stress) Endoplasmic reticulum stress refers to a series of adaptive responses initiated by the endoplasmic reticulum when the protein processing, folding, and transport processes in the endoplasmic reticulum become disordered or overloaded.

[0052] Chemical chaperones are a class of small molecules that can help proteins in cells fold correctly and relieve endoplasmic reticulum stress. 4-phenyl butyric acid (PBA) and tauroursodeoxycholic acid (TUDCA) are two known chaperone compounds that can relieve endoplasmic reticulum stress caused by unfolded proteins.

[0053] This study found that ferroptosis inhibitor-1 can alleviate the toxic effects of tunicamycin on plants.

[0054] The technical solution of the embodiment of the present application includes the use of a ferroptosis inhibitor-1 in the preparation of a tunicamycin phytotoxicity inhibitor, and the structure of the ferroptosis inhibitor-1 is shown in formula (I):

[0055]

[0056] In some embodiments, tunicamycin phytotoxicity inhibitors can be used to alleviate the toxic effects of tunicamycin on plants.

[0057] In some embodiments, the toxic effect of tunicamycin on plants includes at least one of inhibiting seed germination and inhibiting seedling growth.

[0058] In some embodiments, the plant includes at least one of a cruciferous plant, a gramineous plant, a leguminous plant, a solanaceous plant, a cucurbitaceous plant, a rosaceous plant, a rutaceous plant, and an umbelliferous plant.

[0059] A method for alleviating tunicamycin toxicity in plants, comprising:

[0060] Treating plant seeds or seedlings with ferroptosis inhibitor-1;

[0061] The structure of ferroptosis inhibitor-1 is shown in formula (I):

[0062]

[0063] In some embodiments, the method of treating plant seeds or seedlings with ferroptosis inhibitor-1 comprises:

[0064] Soak the roots of plant seeds or seedlings in a solution of ferroptosis inhibitor-1.

[0065] In some embodiments, the method of treating plant seeds or seedlings with ferroptosis inhibitor-1 comprises:

[0066] A plant culture medium containing a ferroptosis inhibitor is prepared, and the plant seeds or seedlings are cultured in the plant culture medium.

[0067] It can be understood that the plant culture medium containing the ferroptosis inhibitor can be completely self-prepared, or it can be semi-prepared by purchasing a commercial plant culture medium and adding a certain proportion of the ferroptosis inhibitor thereto.

[0068] An application of a ferroptosis inhibitor-1 in the preparation of a pesticide composition, the pesticide composition comprising the ferroptosis inhibitor-1 and a pesticide adjuvant;

[0069] The structure of ferroptosis inhibitor-1 is shown in formula (I):

[0070]

[0071] In some embodiments, the pesticide composition can be used to alleviate the toxic effects of tunicamycin on plants.

[0072] In some embodiments, the toxic effect of tunicamycin on plants includes at least one of inhibiting seed germination and inhibiting seedling growth.

[0073] In some embodiments, the plant includes at least one of a cruciferous plant, a gramineous plant, a leguminous plant, a solanaceous plant, a cucurbitaceous plant, a rosaceous plant, a rutaceous plant, and an umbelliferous plant.

[0074] A soil detoxification method comprising:

[0075] Apply ferroptosis inhibitor-1 to the soil and stir;

[0076] The structure of ferroptosis inhibitor-1 is shown in formula (I):

[0077]

[0078] In some embodiments, the method of applying ferroptosis inhibitor-1 to the soil comprises:

[0079] Firstly, the pesticide is prepared, and the composition of the pesticide includes ferroptosis inhibitor-1 and pesticide adjuvant;

[0080] The pesticide is applied on the soil, and the application method includes spraying, pouring and other pesticide application methods commonly used in the agricultural field.

[0081] The present application provides an application of a ferroptosis inhibitor-1 in the preparation of a tunicamycin plant toxicity inhibitor and a pesticide composition, as well as a method for alleviating tunicamycin toxicity in plants and a soil attenuation method.

[0082] The ferroptosis inhibitor-1 of the embodiment of the present application can significantly improve the tolerance of plants to tunicamycin toxicity, and help plants and their seeds to germinate and grow normally under conditions of higher concentrations of tunicamycin.

[0083] In some embodiments, the use of a 1 μM ferroptosis inhibitor-1 solution can alleviate plant growth inhibition and seed death after germination caused by 0.02-0.2 μM tunicamycin treatment.

[0084] The following are some specific embodiments.

[0085] For experimental parameters not specified in the following specific embodiments, reference is made to the instructions given in the present application document, and reference may also be made to experimental manuals in the art or other experimental methods known in the art, or to experimental conditions recommended by manufacturers.

[0086] The raw materials and reagents involved in the following specific examples can be obtained from commercial sources, or can be prepared by those skilled in the art according to known methods.

[0087] The Arabidopsis thaliana bzip28 / 60 double mutant and nrt1.7-2 mutant were constructed by referring to the method described in patent document CN113549645A. The bzip28 / 60 double mutant is a tunicamycin hypersensitive mutant, and the nrt1.7-2 mutant is a tunicamycin insensitive mutant.

[0088] The wild type yeast strain is the INVSc1 wild type strain, and the tunicamycin-hypersensitive yeast strain is a self-constructed strain overexpressing the tunicamycin transporter. The strain characteristics and construction methods are described in the literature: [Liu C, Hao D, Sun R, Zhang Y, Peng Y, Yuan Y, Jiang K, Li W, Wen X, Guo H. Arabidopsis NPF2.13 functions as a critical transporter of bacterial natural compound tunicamycin in plant-microbe interaction. New Phytol. 2023Apr;238(2):765-780.doi:10.1111 / nph.18752.Epub 2023Feb 9.PMID:36653958.]

[0089] Example 1

[0090] This embodiment provides an application of the ferroptosis inhibitor-1 of the present application in the preparation of a tunicamycin phytotoxicity inhibitor, including the following contents:

[0091] (1) Detoxification effect of the small molecule compound to be tested on Arabidopsis thaliana poisoned by tunicamycin

[0092] Experimental culture medium was prepared by adding no small molecule compounds, 1 μM 4-phenylbutyric acid (PBA), taurodeoxycholic acid (TUDCA), ferroptosis inhibitor CPX (CPX) and ferroptosis inhibitor-1 (Fer-1) to 1 / 2MS culture medium containing 20 nM and 50 nM tunicamycin, respectively. The germination and seedling phenotypes of Arabidopsis thaliana bzip28 / 60 double mutant on the experimental culture medium were observed and recorded.

[0093] Figure 1 This is a comparison chart of the detoxification effects of 4-phenylbutyric acid, taurodeoxycholic acid, ferroptosis inhibitor CPX and ferroptosis inhibitor-1 on the Arabidopsis bzip28 / 60 double mutant poisoned by tunicamycin in Example 1 of the present application. Figure 1 TM refers to tunicamycin, mock refers to the blank control group poisoned with tunicamycin without adding any small molecules, PBA refers to the 4-phenylbutyric acid group, TUDCA refers to the taurodeoxycholic acid group, CPX refers to the ferroptosis inhibitor CPX group, and Fer-1 refers to the ferroptosis inhibitor-1 group.

[0094] (2) Detoxification effect of ferroptosis inhibitor-1 on tunicamycin-toxic Arabidopsis

[0095] Experimental culture medium was prepared by adding 0μM, 0.1μM, and 1μM ferroptosis inhibitor-1 (Fer-1) to 1 / 2MS culture medium without tunicamycin and containing 0.2μM tunicamycin, respectively. The germination and seedling phenotypes of the tunicamycin hypersensitive mutant bzip28 / 60 on the experimental culture medium were observed and recorded.

[0096] Figure 2 This is a comparison diagram of the detoxification effects of different concentrations of ferroptosis inhibitor-1 on wild-type Arabidopsis thaliana poisoned by tunicamycin in Example 1 of the present application; Figure 2 A in the middle is a photo comparison of the growth status of wild-type Arabidopsis under different treatment conditions;

[0097] Figure 2 Middle B is a bar chart comparing the relative weights of wild-type Arabidopsis under different treatment conditions. Figure 2 The numerical values ​​under the Fer-1 symbol in A represent different concentrations of ferroptosis inhibitor-1, mock is 1 / 2MS medium without tunicamycin, and 0.2 μM TM is 1 / 2MS medium containing 0.2 μM tunicamycin. Figure 2 The vertical axis of B refers to the relative weight, and the groups represented by the horizontal axis from left to right are 1 / 2MS medium without tunicamycin added with 0μM, 0.1μM, and 1μM ferroptosis inhibitor-1, and 1 / 2MS medium containing 0.2μM tunicamycin added with 0μM, 0.1μM, and 1μM ferroptosis inhibitor-1.

[0098] according to Figure 2 , FIP-1 treatment alone slightly inhibited plant growth, suggesting that normal use of FIP-1 may interfere with cellular redox homeostasis.

[0099] Experimental culture medium was prepared by adding 0μM, 0.1μM, 1μM, and 5μM of ferroptosis inhibitor-1 (Fer-1) to 1 / 2MS culture medium without tunicamycin and containing 0.2μM tunicamycin, respectively. The germination and seedling phenotypes of Arabidopsis wild type, Arabidopsis bzip28 / 60 double mutant, and Arabidopsis nrt1.7-2 mutant on the experimental culture medium were observed and recorded.

[0100] Figure 3 This is a comparison chart of the detoxification effects of different concentrations of ferroptosis inhibitor-1 on tunicamycin-poisoned Arabidopsis wild type, Arabidopsis bzip28 / 60 double mutant and Arabidopsis nrt1.7-2 mutant in Example 1 of the present application. Figure 3The numerical values ​​under the Fer-1 mark in the figure represent different concentrations of ferroptosis inhibitor-1, and the figures under the WT, bzip28 / 60 and nrt1.7-2 marks represent the germination and seedling phenotypes of Arabidopsis wild type, Arabidopsis bzip28 / 60 double mutant and Arabidopsis nrt1.7-2 mutant on the culture medium, respectively, where mock is 1 / 2MS culture medium without tunicamycin, and TM is 1 / 2MS culture medium containing 0.2 μM tunicamycin.

[0101] The results are as follows Figure 3 , the simultaneous treatment of the ferroptosis inhibitor CPX and tunicamycin did not show any reduction or enhancement of the toxicity of tunicamycin. However, the simultaneous application of tunicamycin and the ferroptosis inhibitor-1 significantly inhibited the toxicity of tunicamycin to both the double mutant and the wild-type plants, as shown by normal seed germination and normal development of cotyledons and true leaves of seedlings.

[0102] according to Figure 3 It was also found that the toxic effects of ultra-high concentrations of tunicamycin could not be alleviated by higher concentrations of ferroptosis inhibitor-1, indicating that ferroptosis inhibitor-1 has a certain range of working concentrations and action limits.

[0103] (3) Effects of ferroptosis inhibitor-1 and tunicamycin on UPR gene expression in Arabidopsis

[0104] Seven-day-old wild-type Arabidopsis seedlings were transferred to liquid MS medium containing DMSO, 1 μM ferroptosis inhibitor-1, 0.1 μM tunicamycin, 1 μM ferroptosis inhibitor-1, and 0.1 μM tunicamycin. After 72 hours of treatment, the plants were collected and RNA was extracted. After reverse transcription, qPCR was performed to detect the expression levels of UPR marker genes BiP1 / 2, CNX1, and CRT1.

[0105] Figure 4 The comparison results of UPR gene expression in Arabidopsis wild type poisoned by tunicamycin with and without the addition of ferroptosis inhibitor-1 for detoxification in Example 1 of the present application are as follows: Figure 4 Mock represents the blank control culture medium with DMSO added, and Fer-1, TM, and TM+Fer-1 represent the experimental culture medium with 1 μM ferroptosis inhibitor-1, 0.1 μM tunicamycin, 1 μM ferroptosis inhibitor-1, and 0.1 μM tunicamycin added, respectively. Figure 4 A, B and C in the figure are the expression levels of UPR marker genes BiP1 / 2, CNX1 and CRT1 in Arabidopsis wild-type seedlings cultured in each group of culture media.

[0106] according to Figure 4It can be seen that the upregulation of UPR gene expression caused by tunicamycin treatment was not affected by the addition of ferroptosis inhibitor-1, indicating that tunicamycin can effectively enter the cells and produce toxic effects, suggesting that the detoxification function of ferroptosis inhibitor-1 is not achieved by directly acting on the degradation of tunicamycin or preventing it from entering the cells.

[0107] (4) The mitigating effect of ferroptosis inhibitor-1 on Arabidopsis thaliana poisoned by other ER stress inducers

[0108] Gradient concentrations of dithiothreitol DTT (0 μM, 1.2 μM, 1.5 μM) and proline analog L-azaaziridine-2-carboxylic acid AZC (0 μM, 50 μM, 100 μM) were added to MS medium without ferroptosis inhibitor-1 or with 1 μM ferroptosis inhibitor-1, respectively. Arabidopsis wild-type and Arabidopsis bzip28 / 60 double mutant seeds were cultured in the above medium for 10 days, and their germination and growth phenotypes were recorded by photographing.

[0109] Figure 5 This is a comparative diagram of the detoxification effects of different concentrations of ferroptosis inhibitor-1 on Arabidopsis wild type and Arabidopsis bzip28 / 60 double mutant poisoned by dithiothreitol (DTT) and proline analog L-azaaziridine-2-carboxylic acid (AZC) in Example 1 of the present application. Figure 5 The numerical values ​​under the DTT mark in A represent different concentrations of dithiothreitol, and the graphs under the WT and bzip28 / 60 marks represent the germination and seedling phenotypes of Arabidopsis wild type and Arabidopsis bzip28 / 60 double mutant on the culture medium, respectively, where mock is MS medium without ferroptosis inhibitor-1, and Fer-1 is MS medium containing 1 μM ferroptosis inhibitor-1.

[0110] according to Figure 5 It can be seen that ferroptosis inhibitor-1 cannot alleviate the growth inhibition of seedlings by L-azaaziridine-2-carboxylic acid, and ferroptosis inhibitor-1 aggravates the growth inhibition toxic effect of dithiothreitol on Arabidopsis seedlings.

[0111] Comparative Example 1

[0112] This comparative example studies the detoxification effect of ferroptosis inhibitor-1 on yeast cells poisoned by tunicamycin, and the steps are as follows:

[0113] 1 μM ferroptosis inhibitor-1, 0.2 μM tunicamycin, 0.2 μM tunicamycin and 1 μM ferroptosis inhibitor-1 were added to SG-U culture medium, respectively. Another group of cells to which an equal volume of blank solvent (DMSO) was added was taken as the control group. Wild-type yeast cells were cultured with the above culture medium. After 48 h of culture, OD600 (OD600 refers to the absorbance value of the solution at a wavelength of 600 nm) was measured.

[0114] 1 μM ferroptosis inhibitor-1, 0.02 μM tunicamycin, 0.02 μM tunicamycin and 1 μM ferroptosis inhibitor-1 were added to SG-U culture medium respectively. Another group of cells to which an equal volume of blank solvent (DMSO) was added was taken as the control group. Tunicamycin-hypersensitive yeast cells were cultured with the above culture medium. After 48 h of culture, OD600 was measured.

[0115] Figure 6 This is a comparison chart of the detoxification effect of the ferroptosis inhibitor-1 of Comparative Example 1 of the present application on wild-type and tunicamycin-hypersensitive yeast cells poisoned by tunicamycin; Figure 6 A in the middle is the detoxification effect of ferroptosis inhibitor-1 on wild-type yeast cells poisoned by tunicamycin, mock is the control group with an equal volume of blank solvent (DMSO), Fer-1, TM and TM+Fer-1 are the experimental groups with 1 μM ferroptosis inhibitor-1, 0.2 μM tunicamycin, 1 μM ferroptosis inhibitor-1 and 0.2 μM tunicamycin, respectively; Figure 6 Middle B is the detoxification effect of ferroptosis inhibitor-1 on tunicamycin-hypersensitive yeast cells, mock is the control group with an equal volume of blank solvent (DMSO), Fer-1, TM and TM+Fer-1 are the experimental groups with 1μM ferroptosis inhibitor-1, 0.02μM tunicamycin, 1μM ferroptosis inhibitor-1 and 0.02μM tunicamycin, respectively.

[0116] according to Figure 6 As a result, ferroptosis inhibitor-1 had no obvious detoxification effect on yeast cells poisoned by tunicamycin, and it could not restore the severe inhibition of yeast cell growth by tunicamycin at a high dose (1 μM).

[0117] From the above experimental results, we can see that unlike compound molecular chaperones that are suitable for animal cells, ferroptosis inhibitor-1 is a plant-specific small molecule inhibitor of tunicamycin toxicity.

[0118] All documents mentioned in this application are cited as references in this application, just as each document is cited as reference separately. Unless they conflict with the application purpose and / or technical solution of this application, the cited documents involved in this application are cited with all contents and all purposes. When the cited documents are involved in this application, the definitions of relevant technical features, terms, nouns, phrases, etc. in the cited documents are also cited. When the cited documents are involved in this application, the examples and preferred methods of the cited relevant technical features can also be incorporated into this application as references, but are limited to the implementation of this application. It should be understood that when the cited content conflicts with the description in this application, the present application shall prevail or be modified adaptively according to the description of this application.

[0119] The technical features of the above-mentioned implementation modes and examples can be combined in any appropriate manner. To make the description concise, not all possible combinations of the technical features in the above-mentioned implementation modes and examples are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0120] The above-described embodiments only express several implementation methods of the present application, but they cannot be understood as limiting the scope of the patent application. It should be pointed out that, for those of ordinary skill in the art, without departing from the concept of the present application, several variations and improvements can also be made, which all belong to the protection scope of the present application. In addition, it should be understood that after reading the above-mentioned teaching content of the present application, those skilled in the art can make various changes or modifications to the present application, and the equivalent forms obtained also fall within the protection scope of the present application. It should also be understood that the technical solutions obtained by those skilled in the art on the basis of the technical solutions provided in the present application through logical analysis, reasoning or limited experiments are all within the protection scope of the claims attached to the present application. Therefore, the protection scope of the patent of the present application shall be based on the attached claims, and the description can be used to interpret the content of the claims.

Claims

1. Use of a ferroptosis inhibitor-1 in the preparation of a tunicamycin phytotoxicity inhibitor, wherein the structure of the ferroptosis inhibitor-1 is shown in formula (I):

2. The use according to claim 1, characterized in that The tunicamycin phytotoxicity inhibitor can be used to alleviate the toxic effect of tunicamycin on plants.

3. The use according to claim 2, characterized in that The toxic effect of tunicamycin on plants includes at least one of inhibiting seed germination and inhibiting seedling growth.

4. The use according to claim 2 or 3, characterized in that: The plant comprises at least one of a cruciferous plant, a gramineous plant, a leguminous plant, a solanaceous plant, a cucurbitaceous plant, a rosaceous plant, a rutaceous plant, and an umbelliferous plant.

5. A method for alleviating tunicamycin toxicity in plants, characterized in that: include: treating seeds or seedlings of the plant with ferroptosis inhibitor-1; The structure of the ferroptosis inhibitor-1 is shown in formula (I):

6. Use of a ferroptosis inhibitor-1 in the preparation of a pesticide composition, characterized in that: The pesticide composition comprises the ferroptosis inhibitor-1 and a pesticide adjuvant; The structure of the ferroptosis inhibitor-1 is shown in formula (I):

7. The use according to claim 6, characterized in that The pesticide composition can be used to alleviate the toxic effects of tunicamycin on plants.

8. The use according to claim 7, characterized in that The toxic effect of tunicamycin on plants includes at least one of inhibiting seed germination and inhibiting seedling growth.

9. The use according to any one of claims 6 to 8, characterized in that: The plants include Arabidopsis thaliana.

10. A soil detoxification method, characterized in that: include: applying ferroptosis inhibitor-1 to the soil and stirring; The structure of the ferroptosis inhibitor-1 is shown in formula (I):

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