Use of pyrazolate as an analog of a strigolactone

Abstract

This invention discloses the application of pyrazolite as a strigolactone analog. The pyrazolite of this invention can be used as a strigolactone analog to regulate hypocotyl length, seed germination, root hair length and / or density, and plant height, exhibiting similar effects to the strigolactone analog rac-GR24. Furthermore, pyrazolite is inexpensive, applicable to a variety of plants, and has minimal environmental impact, allowing for large-scale application in agricultural production.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, specifically relating to the application of pyrazolite as a strigolactone analog. Background Technology

[0002] Strigolactones (SLs) are a novel group of plant hormones that exert a wide range of significant effects from seed germination to late-stage plant development. Plant hormones exert their effects in extremely small amounts within the plant, making extraction from plants virtually impossible. Furthermore, the synthesis of SL analogs faces numerous challenges, including high costs, low efficiency, biochemical safety, and chemical stability. Therefore, the application of SLs has remained largely confined to basic laboratory research rather than practical production.

[0003] Using exogenous strigolactone analogs to regulate plant serotonin levels can adjust the basic plant phenotype, bringing the plant shape closer to or even achieving the ideal shape, thereby optimizing yield and quality, resulting in significant economic potential. Furthermore, effective strigolactone analogs can alleviate adverse effects on plants caused by environmental factors such as soil drought, soil salinity stress, and abnormally high temperatures. In today's context of accelerating climate change and increasing extreme weather events, this has enormous and broad application prospects. Therefore, finding a low-cost, highly effective strigolactone analog is of great significance. Summary of the Invention

[0004] The present invention aims to at least solve one of the technical problems existing in the prior art. To this end, the present invention proposes the use of pyrazolite as a strigolactone analogue.

[0005] This invention also proposes a method for regulating the length of the hypocotyl in plants.

[0006] The present invention also proposes a method for promoting plant seed germination.

[0007] The present invention also proposes a method for regulating the length and / or density of plant root hairs.

[0008] This invention also proposes a method for regulating plant height.

[0009] According to a first aspect of the invention, the use of pyrazolite as a strigolactone analogue in any of the following is proposed:

[0010] (1) Regulating the length of the hypocotyl in plants;

[0011] (2) Promotes seed germination in plants;

[0012] (3) Regulating the length and / or density of plant root hairs;

[0013] (4) Regulate plant height.

[0014] In some embodiments of the present invention, regulating the length of the plant hypocotyl includes inhibiting the elongation of the plant hypocotyl.

[0015] In some embodiments of the present invention, the regulation of plant root hair length and / or density includes increasing plant root hair length and / or density.

[0016] In some embodiments of the present invention, the regulation of plant height includes inhibiting or increasing plant height.

[0017] In some embodiments of the present invention, the plant includes cruciferous plants.

[0018] In some embodiments of the present invention, the cruciferous plant includes Arabidopsis thaliana.

[0019] According to a second aspect of the present invention, a method for regulating the length of a plant hypocotyl is provided, the method comprising the steps of: seeding plant seeds in a culture reagent containing pyrazolite and culturing them.

[0020] In some embodiments of the present invention, the concentration of pyrazolite in the culture reagent is 0.01-0.1 µM. For example, it can be 0.01 µM, 0.02 µM, 0.03 µM, 0.04 µM, 0.05 µM, 0.06 µM, 0.07 µM, 0.08 µM, 0.09 µM, or 0.1 µM.

[0021] In some embodiments of the present invention, the cultivation step includes vernalization.

[0022] In some embodiments of the present invention, the plant includes cruciferous plants.

[0023] In some embodiments of the present invention, the cruciferous plant includes Arabidopsis thaliana.

[0024] According to a third aspect of the present invention, a method for promoting the germination of plant seeds is provided, the method comprising the steps of: placing vernalized seeds in a container containing a pyrazolite solution for germination culture.

[0025] In some embodiments of the present invention, the concentration of the pyrazolite solution is 0.01-0.05 µM. For example, it can be 0.01 µM, 0.02 µM, 0.03 µM, 0.04 µM, or 0.05 µM.

[0026] In some embodiments of the present invention, the container includes a petri dish.

[0027] In some embodiments of the present invention, the petri dish further includes filter paper.

[0028] In some embodiments of the present invention, the plant includes cruciferous plants.

[0029] In some embodiments of the present invention, the cruciferous plant includes Arabidopsis thaliana.

[0030] According to a fourth aspect of the present invention, a method for regulating the length and / or density of plant root hairs is provided, the method comprising the steps of: sowing plant seeds in a culture reagent containing pyrazolite and culturing them.

[0031] In some embodiments of the present invention, the concentration of pyrazolite in the culture reagent is 0.01-0.1 µM. For example, it can be 0.01 µM, 0.02 µM, 0.03 µM, 0.04 µM, 0.05 µM, 0.06 µM, 0.07 µM, 0.08 µM, 0.09 µM, or 0.1 µM.

[0032] In some embodiments of the present invention, the cultivation step includes vernalization.

[0033] In some embodiments of the present invention, the cultivation step further includes culturing after vernalization at 20-25°C under conditions of 15-17 hours of light and 6-8 hours of darkness.

[0034] In some embodiments of the present invention, the cultivation step further includes culturing at 22°C under conditions of 16 hours of light and 8 hours of darkness after vernalization.

[0035] In some embodiments of the present invention, the plant includes cruciferous plants.

[0036] In some embodiments of the present invention, the cruciferous plant includes Arabidopsis thaliana.

[0037] According to a fifth aspect of the present invention, a method for regulating plant height is provided, the method comprising the step of applying a solution containing pyrazolite to the plant.

[0038] In some embodiments of the present invention, the effective concentration of pyrazolite in the solution is 1-25 µM.

[0039] In some embodiments of the present invention, the effective concentration of pyrazolite in the solution is 5-20 µM. For example, it can be 5 µM, 6 µM, 7 µM, 8 µM, 9 µM, 10 µM, 12 µM, 13 µM, 14 µM, 15 µM, 16 µM, 17 µM, 18 µM, 19 µM, or 20 µM.

[0040] In some embodiments of the present invention, when regulating plant height to reduce plant height, the effective concentration of pyrazolite in the solution is 5-10 µM. For example, it can be 5 µM, 6 µM, 7 µM, 8 µM, 9 µM, or 10 µM.

[0041] In some embodiments of the present invention, the amount of solution applied is 0.1-3 mL / time / plant.

[0042] In some embodiments of the present invention, the amount of solution applied is 1-3 mL / time / plant.

[0043] In some embodiments of the present invention, the amount of solution applied is 2 mL / time / plant.

[0044] In some embodiments of the present invention, the solution is applied 2-4 times.

[0045] In some embodiments of the present invention, the solution is applied three times.

[0046] In some embodiments of the present invention, the application to the plant is performed after the plant bolts.

[0047] In some embodiments of the present invention, the plant includes cruciferous plants.

[0048] In some embodiments of the present invention, the cruciferous plant includes Arabidopsis thaliana.

[0049] According to some embodiments of the present invention, at least the following beneficial effects are achieved: the pyrazolite of the present invention can be used as a strigolactone analog to regulate the hypocotyl length, seed germination, root hair length and / or density, and plant height, and has similar effects to the strigolactone analog rac-GR24; moreover, pyrazolite is inexpensive, can be applied to a variety of plants, has little environmental pollution, and can be used on a large scale in agricultural production.

[0050] The specific applications in agricultural production can be expected as follows: The pyrazolite provided by this invention has an inhibitory effect on the hypocotyl. Related studies suggest that the inhibition of the hypocotyl can accelerate the formation of photomorphology in seedlings, save energy on the hypocotyl for the growth of roots and young leaves, and in some cases increase the survival rate of seedlings. The regulation of plant height is also a key topic in agricultural production and research, and its influence on plant type further affects crop yield, quality and other factors. Attached Figure Description

[0051] Figure 1 The diagram shows the structural formula and molecular docking analysis results of pyrazolite in the embodiments of the present invention; wherein, A is the molecular structural formula of pyrazolite, and B is the molecular docking result diagram of pyrazolite.

[0052] Figure 2 The diagram shows the effect of pyrazolite on the germination of Arabidopsis thaliana in this embodiment of the invention; where A is the detection result of the change in the germination rate of Arabidopsis thaliana, and B is the result of the change in the germination potential of Arabidopsis thaliana.

[0053] Figure 3 The figure shows the effect of pyrazolite on the length of the hypocotyl in Arabidopsis thaliana according to the embodiments of the present invention, where "**" means p < 0.01;

[0054] Figure 4 The figure shows the effect of pyrazolite on root hair growth in Arabidopsis thaliana according to the embodiments of the present invention. In the figure, A is WT Arabidopsis thaliana; B is max4 mutant Arabidopsis thaliana; C is d14 mutant Arabidopsis thaliana; the scale bar in the figure is 1 μm. Detailed Implementation

[0055] The following will describe the concept and technical effects of the present invention clearly and completely with reference to the embodiments, so as to fully understand the purpose, features and effects of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of them. Other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are all within the scope of protection of the present invention. Unless otherwise specified, the experimental methods used in the embodiments are conventional methods; the materials and reagents used, unless otherwise specified, are commercially available.

[0056] Reagents and materials:

[0057] Preparation of pyrazolite (purchased from Bailingwei Technology Co., Ltd.): Weigh the required mass of the reagent pure using a 0.1 g balance, dissolve it, and add ultrapure water to a final volume of 10 mL to prepare a 10 mM stock solution. Take 400 μL of the 10 mM stock solution and add it to a 2 mL EP tube containing 1.6 mL of ultrapure water to prepare a 2 mM stock solution. Take 40 μL of the 10 mM pyrazolite stock solution and add it to a 2 mL EP tube containing 1.96 mL of ultrapure water to prepare a 0.2 mM stock solution.

[0058] Preparation of rac-GR24 (purchased from Beijing Solarbio Technology Co., Ltd.): Weigh the required mass of the reagent pure product using a 0.1 g balance, dissolve it, add ultrapure water to make up to 10 mL, and prepare a 10 mM stock solution.

[0059] The relative molecular masses of pyrazolite and rac-GR24 reagents and the required mass are shown in Table 1 below.

[0060] Table 1

[0061]

[0062] Preparation of solid MS culture medium:

[0063] For the half-solid MS medium used in the experiment, analytical grade sucrose (purchased from Sinopharm Chemical Reagent Co., Ltd.), analytical grade agar powder (purchased from Beijing Solarbio Science & Technology Co., Ltd.), and MS medium base salts (purchased from Beijing Cooler Technology Co., Ltd.) were weighed according to the proportions shown in Table 2. The mass of each component weighed in the experiment was reduced proportionally according to the volume used. During preparation, the sucrose and MS powder were dissolved first. The pH of the sucrose-MS powder mixture was adjusted to 5.8-6.0 using 1% KOH solution before adding the agar powder, shaking well, sealing, and waiting for steam sterilization before use.

[0064] Table 2. Partition ratios of each group in 1L 1 / 2 solid MS medium

[0065]

[0066] The Arabidopsis thaliana seeding and vernalization in this embodiment were performed according to the following steps:

[0067] 1) Take the conical flask, wash it and rinse it with pure water. Measure the required amount of ultrapure water into it using a graduated cylinder. Weigh the required amount of medicine using a 0.1% balance, and seal the flask with a sealing film after dissolving and adjusting the pH.

[0068] 2) Take the required number of petri dishes for the experiment, wash and dry them, and then put them into a steam sterilizer along with the items to be sterilized (petri dishes, culture medium, pipette tips, ultrapure water, 0.1% agar). After sterilization, place the petri dishes, pipette tips, and culture medium in a 65℃ oven to dry or keep warm for later use.

[0069] 3) Place the petri dish, pipette, pipette tip, sealing film, ultrapure water, 0.1% agar, 75% ethanol, and marker into the laminar flow hood. After UV sterilization for 45 minutes, ventilate for 15 minutes. Light the alcohol lamp in the laminar flow hood and begin the inoculation process.

[0070] 4) Remove the culture medium from the oven, let it cool slightly, then add the corresponding pyrazolite or GR24 stock solution to the culture medium, mix thoroughly, pour into a culture dish with the label written on the bottom, and let it cool for later use.

[0071] 5) Take an appropriate amount of Arabidopsis seeds into a 2 mL EP tube, soak them in 75% ethanol for 10 min for disinfection, remove the ethanol after disinfection, rinse with ultrapure water 4-5 times, and then add an appropriate amount of 0.1% agar to the Arabidopsis seeds until the mixture can be easily and smoothly aspirated with a 1000 μL pipette. This completes the seed treatment.

[0072] 6) Using a 1000 μL pipette tip, take an appropriate amount of Arabidopsis seed mixture. Remove the pipette tip containing the seeds and use it to evenly sow the Arabidopsis seeds one by one onto the completely solidified culture medium. After filling the entire medium, leave the petri dish uncovered and set it aside to allow the 0.1% agar moisture added with the seeds to evaporate. Continue sowing other treated petri dishes in a similar manner.

[0073] 7) After the 0.1% agar has completely evaporated, place the petri dish over the flame of an alcohol lamp, scorch the mouth of the dish with the flame, and then seal the petri dish with sealing film.

[0074] 8) After sealing all petri dishes and checking the sealing effect a second time, place the sealed petri dishes in a 4°C constant temperature refrigerator for vernalization for 2 days. This completes the inoculation process.

[0075] Example 1

[0076] This embodiment provides the application of pyrazolite as a strigolactone analogue, and the specific screening and verification process is as follows:

[0077] 1. Structural characteristics analysis of pyrazolite

[0078] The molecular formula of pyrazolite is C 19 H 16 Cl2N2O4S is an agricultural herbicide developed by Sankyo Chemical Co., Ltd. in the 1970s. It is a type of hydroxyphenylpyruvate dioxygenase inhibitor that affects the synthesis of plastoquinones and tocopherols in some plants, thereby affecting the growth of some plants.

[0079] This invention, through extensive screening and research, identified pyrazolite, a structural analog of SLs, from an organic molecular library. Molecular docking tests were performed between pyrazolite and the D14 protein using AutoDock Vina, with results as follows: Figure 1 As shown, from Figure 1 As can be seen from the results, the binding energy of the optimal binding conformation among multiple docking results is -7.72 kcal / mol. In contrast, the optimal binding energy of the GR24-D14 molecular docking test is -8.34 kcal / mol. It is generally believed that the lower the binding energy value, the higher the feasibility of binding. If the optimal conformations of GR24, pyrazolite and D14 are simultaneously displayed in the same molecular model diagram, it can be observed that the binding sites of the two small molecules with the D14 protein are very close.

[0080] In summary, it can be inferred that pyrazolite may bind to the D14 protein, which is an extremely important protein in the SL signaling pathway. Pyrazolite may exert its effects at the SL level by interacting with the D14 protein. Therefore, further verification of pyrazolite is needed.

[0081] 2. Effects of pyrazolite on seed germination in Arabidopsis thaliana

[0082] Experimental plan:

[0083] Treatment of Arabidopsis thaliana seeds: Take an appropriate amount of WT Arabidopsis thaliana seeds, place them in an EP tube, add ultrapure water and shake to submerge the seeds, then place them in a 4℃ refrigerator for vernalization for 2 days. Dissolve 3.5 μL of 100 μM pyrazolite stock solution and 17.5 μL of 2 mM rac-GR24 stock solution in 7 mL of ultrapure water to prepare treatment solutions, i.e., pyrazolite concentration of 0.05 μM and rac-GR24 concentration of 5 μM. Pour the treatment solutions into petri dishes lined with filter paper (for the blank control group, directly pour in 7 mL of ultrapure water), and spread the vernalized Arabidopsis thaliana seeds on the filter paper, ensuring the seeds are completely submerged. Place 50 seeds in each treatment. After labeling, place in a seedling room at 22℃ in the dark for 24 hours to await germination.

[0084] Germination rate and germination potential were calculated as follows: Seeds were photographed, and the number of germinated seeds was counted from the photos to calculate the germination rate. Photos were taken at 0 h, 24 h, 29 h, 34 h, and 37 h after the Arabidopsis thaliana was placed in the seedbed. Data were collected and plotted for analysis after the experiment was completed. The experiment was repeated three times.

[0085] Germination rate / % = (Number of germinated Arabidopsis seeds / Total number of Arabidopsis seeds that eventually germinated normally) × 100%.

[0086] Germination potential / % = (Number of Arabidopsis seeds germinating within the time period / Total number of Arabidopsis seeds that eventually germinate normally) × 100%.

[0087] as follows Figure 2 As shown in Figure A, after 24 h and 29 h of treatment with Arabidopsis seeds, the germination rates of the pyrazolite and rac-GR24 treatments were both higher than those of the blank control group. At 29 h, the germination rate of the pyrazolite treatment was 41.5%, which was 6.3% higher than the 35.2% of the blank control, indicating that its promoting effect on germination was slightly less than that of rac-GR24. At 34 h, the germination rates of the blank control group were similar to those of the two treatment groups. However, at 37 h, the germination rate of the blank control group exceeded that of both treatment groups.

[0088] As can be seen from Figure B, in terms of germination potential, in the early stage of seed germination (0~29 h), the germination potential of Arabidopsis treated with pyrazolite and rac-GR24 was greater than that of the blank control group; when the time reached 34 h, the germination rate of the blank control group was greater than that of the two treatment groups.

[0089] The results showed that treatment with rac-GR24 and pyrazolite could promote the early germination of Arabidopsis seeds, a phenomenon similar to that observed with the treatment of Strelitzia seeds with GR24 in related technologies.

[0090] Treatment with 0.05 μM pyrazolite promoted early germination of Arabidopsis seeds, and the germination potential of the seeds 24-29 h after treatment was similar to that of rac-GR24. This indicates that 0.05 μM pyrazolite also has the function of breaking seed dormancy.

[0091] 3. Effect of pyrazolite on hypocotyl length in Arabidopsis thaliana

[0092] Experimental Methods: Four sterilized 150 mm × 25 mm glass petri dishes and four 60 mL aliquots of half-solid MS medium were prepared. For Arabidopsis seedling cultivation, one control group and three experimental groups containing different concentrations of pyrazolium chloride (0.01, 0.03, and 0.05 μM) were set up. 60 μL, 180 μL, and 300 μL of 10 mM pyrazolium chloride stock solution were added to half-solid MS medium to prepare the experimental group media. For the control group, half-solid MS medium without added reagents was poured into the petri dishes. (The text then abruptly shifts to a discussion of WT and...) max4 Four mutant Arabidopsis species were seeded and vernalized, and then cultured in petri dishes at 22°C under low light conditions of 16 h light and 8 h darkness.

[0093] Hypocotyl length was measured 7 days after Arabidopsis thaliana cultivation. Normally growing Arabidopsis thaliana seedlings were selected and, according to treatment method and species, were evenly and parallelly laid on a black agar background plate with a 1 cm reference frame. The seedlings were photographed using a Canon EOS 5DsR camera (this model is used throughout the text) with a MACRO LENS EF 100 mm f / 2.8L IS USM macro lens (unless otherwise specified). During photography, a tripod was used to ensure the camera's focal plane was parallel to the plane of the seedling. After photography, the hypocotyl length of each seedling was measured using the path measurement function of Digmizer 6 software. The experiment was repeated three times.

[0094] The results are as follows Figure 3 As shown, in WT Arabidopsis treated with pyrazolite, it was observed that compared with the untreated blank control group, the hypocotyl length of Arabidopsis treated with pyrazolite was inhibited and significantly shortened. However, the inhibitory effect of pyrazolite on the hypocotyl length of WT Arabidopsis did not show a concentration-dependent effect within the concentration gradient set in the experiment.

[0095] observe max4 The results from the experimental and control groups of the mutant Arabidopsis thaliana show that... max4 The hypocotyl of the mutant Arabidopsis thaliana control group was significantly longer than that of the wild-type control group, while after treatment with pyrazolium, max4 The hypocotyl length of the mutants was shortened in both cases. Specifically, the two mutants with pyrazolite concentrations of 0.01 μM and 0.03 μM showed this effect. max4 The hypocotyl length in the mutant experimental group was slightly shorter than that in the WT control group. Similar to the WT case, pyrazolite... max4 The inhibitory effect of the mutant Arabidopsis thaliana on hypocotyl length did not show a concentration-dependent effect within the experimental concentration gradient.

[0096] Further analysis of the above experimental results showed that pyrazolium's inhibition of the hypocotyl in *Strombocytoplasmia wt.* (WT) was similar to the results obtained from previous treatment of *Strombocytoplasmia wt.* with rac-GR24, suggesting that pyrazolium may have a similar function to SLs. max4 The experimental results of the mutants also provide evidence for this hypothesis. max4 The mutant itself cannot synthesize SLs normally, but the signal transduction pathway is normal. Exogenous addition of SLs can restore the WT phenotype in Arabidopsis to some extent. The experimental results confirm this; after treatment with pyrazolium, max4 The hypocotyl was suppressed to a length similar to that of WT.

[0097] 4. Effects of pyrazolite on root growth of Arabidopsis thaliana seedlings

[0098] Experimental Methods: Three sets of 90 mm × 15 mm high-transparency quartz glass culture dishes and three 30 mL aliquots of 1 / 2 solid MS medium were prepared. A blank control group, a pyrazolite-treated experimental group, and a rac-GR24 positive control group were set up. The pyrazolite concentration in the experimental group was 0.05 μM, and the concentration of the SLs analog rac-GR24 used in the positive control was 5 μM. 7.5 μL of 0.2 mM pyrazolite stock solution and 75 μL of 2 mM rac-GR24 reagent stock solution were added to 1 / 2 solid MS medium to prepare the experimental group and positive control medium, respectively. For the blank control, the 1 / 2 solid MS medium without reagents was poured onto a plate. For WT and... d14 , max4 Three mutant Arabidopsis species were seeded and vernalized, and then cultured in petri dishes at 22°C under 16 h light and 8 h dark conditions.

[0099] After 10 days of cultivation, the roots of Arabidopsis seedlings were observed. The Arabidopsis seedlings were carefully removed from the culture medium, ensuring the integrity of the root system as much as possible. The remaining agar on the roots was washed off with clean water, and the seedlings were placed on a black background board (the black background board was an agar board with black ink added to keep the roots moist). The growth status of the roots of each Arabidopsis plant was roughly observed, and representative Arabidopsis plants were selected. The roots of these selected Arabidopsis plants were spread out as much as possible so that they did not overlap.

[0100] Observation of root hair growth: The roots of Arabidopsis seedlings were prepared into temporary slides and observed under a regular optical microscope. The differences in root hairs in the root tip maturity zone between different treatments were observed, and the observation results were photographed. The experiment was repeated 3 times.

[0101] Root hair growth status as follows Figure 4 As shown, pyrazolite is effective against WT and d14 , max4 The mutants showed a promoting effect on both the number and length of root hairs. After treatment with 0.05 μM pyrazolite, the number of root hairs in the root tip maturation zone of these three Arabidopsis species increased compared with the untreated blank control group, and the length of the root hairs was also slightly longer than that of the blank control group. However, the effect of pyrazolite treatment on both the increase in the number and the elongation of root hairs was lower than that produced by 5 μM rac-GR24 treatment.

[0102] The results showed that both 0.05 μM pyrazolite and 5 μM rac-GR24 increased the number and length of root hairs, although rac-GR24 had a more significant effect. These results suggest that pyrazolite may have a similar effect to rac-GR24.

[0103] 5. Effects of pyrazolite on Arabidopsis plant height

[0104] Experimental methods:

[0105] Arabidopsis thaliana cultivation: Take WT and d14 , max2 , max4 Four mutant Arabidopsis thaliana seeds were directly sown in untreated 1 / 2 solid MS medium. After vernalization, the medium was transferred to a nursery (22℃ room temperature, 16 h light, 8 h dark) for growth. When the Arabidopsis thaliana reached the 4-6 leaf stage (about 7 days later), the seedlings were transplanted into 10 cm × 10 cm pots to continue growing, with 5 seedlings planted in each pot.

[0106] Preparation of treatment solutions: The experiment will use two reagents, pyrazolite and rac-GR24, to treat Arabidopsis thaliana. The concentrations of pyrazolite treatment solutions are 5, 10, and 20 μM, respectively; the concentration of GR24 treatment solution is 10 μM. 40, 80, and 160 μL of 2 mM pyrazolite stock solution were respectively added to deionized water and brought to a final volume of 16 mL to prepare 5, 10, and 20 μM pyrazolite treatment solutions; 80 μL of rac-GR24 stock solution was added to deionized water and brought to a final volume of 16 mL to prepare a 10 μM GR24 treatment solution.

[0107] Arabidopsis thaliana material treatment: After transplanting, Arabidopsis thaliana plants were cultivated for another 15 days until bolting began. Arabidopsis thaliana plants with similar growth were selected, and four plants were grouped into one group (one pot). WT and max2 The mutant plants were treated with deionized water to serve as a control group. d14 and max4 Both mutants were used as experimental groups and treated with either pyrazolite or rac-GR24. Using a 1000 μL pipette, 2 mL of the treatment solution was added dropwise from the center of the Arabidopsis plant, and each plant was labeled. On days 3 and 8 after the first treatment, the second and third treatments were applied to each type of Arabidopsis in the same manner. After treatment, the four types of Arabidopsis were cultured in a nursery at 22℃ with a 16-hour light-8-hour dark cycle. During this period, the potting soil was kept relatively moist, and watering was done by dripping small amounts from the four corners of the pots. Ten days after the first treatment, the plant height was measured and the average value was taken.

[0108] Table 3. Plant height of Arabidopsis thaliana mutants

[0109]

[0110] Table 3 shows the results of treating Arabidopsis thaliana with pyrazolite under the current experimental design. As can be seen from the table, under the current experimental design, the treatment of Arabidopsis thaliana with pyrazolite resulted in significant improvements in the efficacy of the treatment. max4 After treatment with 5, 10, and 20 μM pyrazolite, the plant heights of the mutants at the first two concentrations were similar and slightly smaller than those of the blank control group. The plant heights of the mutants treated with 20 μM pyrazolite increased by about 20.6%. In the positive control, the plant heights of the mutants treated with 10 μM rac-GR24 increased by about 20.6%.

[0111] In the d14 After the mutants were treated with the solution, their plant height decreased to varying degrees. The plant height reduction after treatment with 10 μM rac-GR24, 5 μM pyrazolite, 10 μM pyrazolite, and 20 μM pyrazolite was 5.1%, 18.6%, 8.7%, and 2.8%, respectively.

[0112] In summary, the results of pyrazolite treatment on various types of Arabidopsis thaliana were highly similar to those of the rac-GR24 positive control group, indicating that pyrazolite is also an analog of SLs.

[0113] The embodiments of the present invention have been described in detail above. However, the present invention is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the spirit of the present invention. Furthermore, the embodiments of the present invention and the features thereof can be combined with each other unless otherwise specified.

Claims

1. The use of pyrazolite in any of the following: (1) Inhibits the elongation of the hypocotyl in plants; (2) Increase the length and / or density of plant root hairs; The plant in question is Arabidopsis thaliana; The inhibition of hypocotyl elongation in plants is achieved through the following steps: planting plant seeds in a culture reagent containing pyrazolite and culturing them; the concentration of pyrazolite in the culture reagent is 0.01-0.1 µM; The increase in plant root hair length and / or density is achieved through the following steps: planting plant seeds in a culture reagent containing pyrazolite and culturing them; the concentration of pyrazolite in the culture reagent is 0.01-0.1 µM.

2. A method for inhibiting hypocotyl elongation in plants, characterized in that, The method includes the following steps: planting plant seeds in a culture reagent containing pyrazolite and culturing them; the concentration of pyrazolite in the culture reagent is 0.01-0.1 µM; The plant in question is Arabidopsis thaliana.

3. A method for increasing the length and / or density of plant root hairs, characterized in that, The method includes the following steps: planting plant seeds in a culture reagent containing pyrazolite and culturing them; the concentration of pyrazolite in the culture reagent is 0.01-0.1 µM; The plant in question is Arabidopsis thaliana.

Images