Inhibition compound taking urine nigatolic acid solanesyl transferase as molecular target and application of inhibition compound

Abstract

The invention relates to the fields of agricultural chemistry and environmental protection, in particular to an inhibition compound with urine black solanesyl transferase (HST) as a molecular target and application of the inhibition compound. An inhibition compound taking urine black acid solanesyl transferase as a molecular target is characterized in that the compound is a compound shown in the following structure, and in the formula, R1, R2, R3 and R4 are respectively Br. The compound has a remarkable inhibition effect on the growth of phytoplankton at low concentration, and shows a powerful weeding effect on weeds such as water barnyard grass and purslane. AlphaFold predictive analysis further confirms that the compounds are combined with HST to inhibit synthesis of plastidoquinone, so that photosynthesis is effectively interfered. The method can be applied to algae and weed control in agriculture and aquaculture, and has a wide application prospect. # imgabs0 #

Description

Technical Field

[0001] The invention relates to the fields of agricultural chemistry and environmental protection, and in particular to an inhibitory compound taking homogentisate solanesyltransferase (HST) as a molecular target and application thereof. Background Art

[0002] Homogentisate solanesyltransferase (HST) is a new potential molecular target for herbicides, which has important application value in improving the killing efficiency and dealing with weeds that are highly resistant to existing chemicals. HST plays a key role in plant biosynthesis, catalyzing the prenylation and decarboxylation of homogentisate to generate 2-methyl-6-solanesyl-1,4-benzoquinol, which is the first intermediate in the biosynthesis of plastoquinone. As an important compound in the electron transport chain of the photosynthetic system, plastoquinone directly affects the photosynthesis of plants. By inhibiting the activity of HST, the synthesis of plastoquinone can be effectively blocked, thereby interfering with the normal photosynthesis. This inhibition not only leads to growth stagnation of plants, but also induces bleaching and eventually leads to their death. Therefore, the development of a compound that inhibits HST provides a new strategy for algae and weed control in environments such as agriculture and aquaculture. Summary of the invention

[0003] The present invention aims to provide an inhibitory compound with homogentisate solanesyltransferase (HST) as a molecular target and application thereof.

[0004] To achieve the above purpose, the present invention adopts the following technical solution:

[0005] An inhibitory compound with homogentisate solanesyl transferase as a molecular target, the compound is a compound shown in the following structure,

[0006]

[0007] Wherein, R1, R2, R3 and R4 are Br respectively.

[0008] An application of the inhibitory compound with homogentisate solanesyl transferase as a molecular target, and an application of the compound in inhibiting homogentisate solanesyl transferase.

[0009] An application of the inhibitory compound with homogentisate solanesyl transferase as a molecular target: application of the compound in the preparation of plant and phytoplankton inhibitors.

[0010] The compound is used in preparing growth inhibitors of Echinochloa crusgalli, Purslane and diatom.

[0011] An inhibitor of homogentisate solanesyl transferase, comprising the compound.

[0012] The concentration of the inhibitory compound solution (aqueous solution) is 3-5 mg / L.

[0013] An application of the inhibitor of homogentisate solanesyl transferase, and the application of the inhibitor in inhibiting the growth of plants and phytoplankton.

[0014] An application of the inhibitor, wherein the inhibitor is sprayed onto plants to be treated, with a minimum application amount of 50 mg / L.

[0015] The advantages of the present invention are:

[0016] The biphenyl compound of the present invention inhibits the synthesis of plastoquinone by binding with HST, thereby hindering the electron transfer process of photosynthesis, thereby effectively interfering with photosynthesis. The compound has a significant growth inhibition effect on phytoplankton at low concentrations, and exhibits a strong weed control effect on weeds such as water barnyard grass and purslane. The invention can be applied to algae and weed control in agriculture and aquaculture, and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 The compound provided in the embodiment of the present invention and the structural prediction of homogentisic acid solanesyltransferase (HST) and the effect diagram of its substrate docking; wherein, A is the three-dimensional structure of HST constructed using ColabFold; B is a docking study performed by AutoDock showing that homogentisic acid binds to HST. The amino acids in the active site of HST are as follows: LEU is leucine, GLY is glycine, VAL is valine, and TYR is tyrosine. Homoditidic acid docks inside the active site and interacts with these amino acids in different ways; C. shows the binding of three structurally similar molecules (i.e., TBB-BP, 2,2'-dihydroxybiphenyl, and 3,3',5,5'-tetrabromobiphenyl) to HST.

[0018] Figure 2 The inhibition rate of different concentrations of the compound TBB-BP provided in the embodiment of the present invention after treating phytoplankton over time

[0019] Figure 3 The inhibition rate of different concentrations of the compound 2,2'-dihydroxybiphenyl provided in the embodiment of the present invention after treating phytoplankton over time

[0020] Figure 4 The inhibition rate of different concentrations of the compound 3,3',5,5'-tetrabromobiphenyl provided in the embodiment of the present invention after treating phytoplankton over time

[0021] Figure 5 This is the maximum photosynthetic efficiency Fv / Fm response curve over time after treating phytoplankton with different concentrations of the compound TBB-BP provided in the embodiment of the present invention.

[0022] Figure 6 This is the plant growth condition of Portulaca oleracea and Echinochloa crusgalli after being treated with different concentrations of the compound TBB-BP provided in the examples of the present invention. DETAILED DESCRIPTION

[0023] The specific implementation modes of the present invention are further described below in conjunction with examples. It should be noted that the specific implementation modes described here are only for illustrating and explaining the present invention, and are not limited to the present invention.

[0024] The compound TBB-BP used in the following examples can be prepared as described in CN108684665B or can also be prepared by conventional chemical synthesis.

[0025] Example 1

[0026] Interaction of compounds with homogentisate solanesyltransferase

[0027] The full length of the plant homogentisate solanesyltransferase HST coding gene was obtained. The AlphaFold prediction technology was used to perform structural analysis on the interaction between the compound (TBB-BP) represented by the structural formula of the present invention and the HST protein.

[0028] The homogentisic acid solanesyltransferase HST gene sequence (NCBI reference sequence: XM_002178060.1) was obtained from plant genome data, and its complete sequence was determined using the Takara SMARTer RACE 5' / 3' kit (TaKaRa). The complete open reading frame of HST is 1164 bp in length, including a putative transit peptide sequence of 22 amino acids. The three-dimensional structure of the HST sequence was constructed using ColabFold, and the normal substrate of HST, homogentisic acid (i.e., the substrate of HST) and the present compound were docked to HST using AutoDock. In order to ensure the accuracy of the docking prediction, two molecules with similar structures to the present compound (i.e., 2,2'-dihydroxybiphenyl and 3,3',5,5'-tetrabromobiphenyl) were used as controls, and their binding sites with HST were determined according to the docking method of the present compound. All structures were visualized using PyMOL (http: / / www.pymol.org) (see Figure 1 AC).

[0029] Depend on Figure 1As shown in A and 1B, the present compound TBB-BP has the same active site as homogentisic acid. The LEU_203, TYR_207, LEU_243 and GLY_246 regions of HST are potential active sites of homogentisic acid, and these amino acids interact with homogentisic acid through hydrogen bonds and hydrophobic bonds. The present compound TBB-BP can interact with three amino acids (i.e., LEU_203, LEU_243 and GLY_246) therein through two halogen bonds, two hydrophobic bonds and one hydrogen bond, and has a site competitive ability similar to homogentisic acid. The results show that 4-BP can compete with the HST active site of homogentisic acid, thereby inhibiting the subsequent biosynthesis of plastoquinone-9.

[0030] To further verify the target site of TBB-BP, the same analysis was performed using two molecules with similar structures to the present compound TBB-BP, namely 2,2'-dihydroxybiphenyl and 3,3',5,5'-tetrabromobiphenyl. Although they also have the potential to interact with the above sites, they only bind to two amino acids. 2,2'-dihydroxybiphenyl interacts with LEU_203 and TYR_207 through hydrophobic bonds and hydrogen bonds, while 3,3',5,5'-tetrabromobiphenyl interacts with LEU_203 and LEU_243 through hydrophobic bonds and halogen bonds. It is the weakened interaction ability of 2,2'-dihydroxybiphenyl and 3,3',5,5'-tetrabromobiphenyl with the homogentisate binding site that makes them unable to compete sufficiently with homogentisate and thus fail to inhibit phytoplankton growth. These data indicate that TBB-BP is able to compete with the HST active site of homogentisate.

[0031] Example 2

[0032] The inhibitory effect of these three molecules with similar structures on phytoplankton was further studied. Specifically, different concentrations of TBB-BP, 2,2'-dihydroxybiphenyl and 3,3',5,5'-tetrabromobiphenyl prepared in water were added to the exponential growth period of phytoplankton P. triangularis CCMP 2561 culture, and the growth dynamics of phytoplankton were observed. The phytoplankton culture without these compounds was used as a control. The changes in fluorescence intensity in the culture system were measured every day (excitation and emission wavelengths were 440nm and 680nm, respectively), and the inhibition rate was calculated, that is, inhibition rate = (FC-FT) / FC×100%, where FT and FC represent the fluorescence intensity of the phytoplankton culture solution with and without the addition of compounds, respectively.

[0033] Depend on Figure 2It can be seen that at a concentration of 0-0.125 mg / L, the compound TBB-BP did not significantly inhibit phytoplankton, but at a concentration higher than 0.5-5 mg / L, the inhibition rate was higher than 80% at 72 hours, indicating that high concentrations of the compound significantly inhibited phytoplankton growth. At a concentration of 0.25 mg / L, although the compound also inhibited phytoplankton growth, the inhibition rate was 40% at 72 hours. In summary, a certain concentration of TBB-BP can inhibit phytoplankton growth.

[0034] Depend on Figure 3 It can be seen that within the tested concentration range (0-13.5 mg / L), the compound 2,2'-dihydroxybiphenyl did not significantly inhibit phytoplankton, indicating that the compound 2,2'-dihydroxybiphenyl did not have an inhibitory effect on phytoplankton growth.

[0035] Depend on Figure 4 It can be seen that within the tested concentration range (0-10 mg / L), compound 3,3',5,5'-tetrabromobiphenyl did not significantly inhibit phytoplankton, indicating that compound 3,3',5,5'-tetrabromobiphenyl did not have an inhibitory effect on phytoplankton growth.

[0036] Depend on Figure 1 It can be seen from C that the interaction between TBB-BP and similar compounds and homogentisate solanesyltransferase HST in the above Example 1, the weaker effect of similar compounds makes 2,2'-dihydroxybiphenyl and 3,3',5,5'-tetrabromobiphenyl have a weaker effect on the homogentisate binding site, so that when they interact with phytoplankton HST and homogentisate, they cannot occupy the action site of homogentisate, and thus cannot inhibit the function of HST, and thus the growth of phytoplankton will not be inhibited. The binding ability of the compound of the present invention to the homogentisate binding site is equivalent to that of homogentisate to its site, and can compete with the HST active site of homogentisate, thereby inhibiting the function of HST and inhibiting the growth of phytoplankton.

[0037] At the same time, the inhibitory effect of the compound of the present invention on phytoplankton was further tested, and the common phytoplankton triangular brown finger algae CCMP 2561 was selected as the test plant. Conventional culture was carried out using f / 2 culture medium at a temperature of 20°C, an illumination of 60μmol m-2s-1, and a light-to-dark cycle of 12h:12h. Phytoplankton in the logarithmic growth period was used as the treatment object. Different concentrations of the compound TBB-BP were added to the above-mentioned treatment objects, and the final concentrations of the compound in the culture solution were 0mg / L, 0.25mg / L, 0.5mg / L, 1mg / L, 1.8mg / L, 2.5mg / L, 5mg / L and 10mg / L, respectively. The solvent DMSO was used as the control group, and then cultured according to the above-mentioned culture conditions, and the number of cells and the photosynthetic parameter Fv / Fm of the cells at 0h, 24h, 48h and 72h were measured respectively. The cells were fixed with glutaraldehyde and then counted by flow cytometry. The photosynthesis of phytoplankton was analyzed and evaluated by chlorophyll fluorescence parameters, and the chlorophyll fluorescence Fv / Fm was measured using the chlorophyll fluorescence instrument WATER-PAM. The maximum photosynthetic efficiency of photosynthetic system II Fv / Fm = (Fm-F0) / Fm is the value measured after 20 minutes of dark adaptation (see Figure 5 ).

[0038] Depend on Figure 5 It can be seen that at concentrations below 1 mg / L, Fv / Fm had no significant difference from the control group, but at 3.0 mg / L and 5.0 mg / L, Fv / Fm dropped to 0, indicating that high concentrations of compounds significantly inhibited photosynthesis. At concentrations of 1.0 mg / L and 1.8 mg / L, although the values ​​of Fv / Fm decreased, they rebounded after 8 hours, and had no significant difference from the control group at 48 hours and 72 hours.

[0039] In summary, a certain concentration of TBB-BP can compete with the HST active site of homogentisate, thereby inhibiting the function of HST. Since HST is one of the key proteins in the plastoquinone pathway, an important compound in the electron transport chain of the synthetic photosynthetic system, the inhibition of HST blocks the synthesis of plastoquinone, directly affecting the photosynthetic electron transport of plants, and the photosynthetic parameter Fv / Fm drops to 0, which impairs plant photosynthesis and inhibits the growth of phytoplankton.

[0040] Example 3

[0041] The inhibitory effect of compounds on weeds

[0042] Common weeds such as water barnyard grass and purslane were used as research objects to test the inhibitory effect of the compounds on weeds. Water barnyard grass and purslane seeds were planted at room temperature and cultured for 4 days according to the conventional cultivation method. The experimental design included foliar spraying treatments. The compounds were prepared with water and the concentrations were set to 1mg / L, 5mg / L and 50mg / L. The foliar spraying was performed when the weeds grew from seeds to seedlings for 4 days. The herbicidal effect of the compound was evaluated by observing the degree of whitening of the plants and the growth status of the plants 2 days after spraying (see Figure 6 ).

[0043] Depend on Figure 6 It can be seen that the compound at 50 mg / L can cause problems such as yellowing of plant leaf tips and plant failure to stand upright, and plant growth stagnation within 72 hours after spraying. The effect of foliar spraying is significant, indicating that the compound can more effectively enter the plant body through the leaves to exert its effect.

[0044] In summary, the compounds of the present invention can inhibit homogentisate solanesyltransferase, and thus can be further used as plant growth inhibitors, and have broad application prospects.

Claims

1. An inhibitory compound with homogentisate solanesyl transferase as a molecular target, characterized in that: The compound is a compound represented by the following structure, Wherein, R1, R2, R3 and R4 are Br respectively.

2. Use of an inhibitory compound with homogentisate solanesyl transferase as a molecular target according to claim 1, characterized in that: The compound is used in inhibiting homogentisate solanesyl transferase.

3. Use of the inhibitory compound with homogentisate solanesyl transferase as a molecular target according to claim 1, characterized in that: The compound is used in preparing plant and phytoplankton inhibitors.

4. The use of the inhibitory compound with homogentisate solanesyl transferase as the molecular target according to claim 1, characterized in that: The compound is used in preparing growth inhibitors of Echinochloa crusgalli, Purslane and diatom.

5. An inhibitor of homogentisate solanesyl transferase, characterized in that: The inhibitor comprises the compound according to claim 1.

6. The inhibitor of homogentisate solanesyl transferase according to claim 5, characterized in that: The concentration of the compound solution in the inhibitor is 3-5 mg / L.

7. A use of the inhibitor of homogentisate solanesyl transferase according to claim 5, characterized in that: The inhibitor is used in inhibiting the growth of plants and phytoplankton.

8. A use of the inhibitor according to claim 7, characterized in that: The inhibitor is sprayed onto the plants to be treated, with the minimum application amount being 50 mg / L.

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