Application of pyrazolate as strigolactone analogue

Abstract

The invention discloses an application of pyrazolate serving as a strigolactone analogue. The pyrazolate provided by the scheme of the invention can be used as the strigolactones analogue for regulating the length of plant hypocotyl, regulating the germination of plant seeds, regulating the length and / or density of plant root hair and regulating the plant height, and has a similar effect with a strigolactones analogue rac-GR24; and pyrazol is low in price, can be applied to various plants, is small in environmental pollution, and can be applied to agricultural production on a large scale.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biotechnology, and particularly relates to application of pyrazoles as strigolactone analogs. BACKGROUND

[0002] Strigolactones (SLs) are a new plant hormone, which has a wide and significant influence on plant seed germination and later development of plants. The plant hormone itself is extremely small in content and plays a role in the plant body, so it is almost not feasible to extract it from plants. At present, the artificial synthesis of SLs analogs has many problems such as high synthesis cost, low efficiency, biochemical safety, chemical stability and the like, so the related application of SLs has been limited to basic research in the laboratory, and has not been put into actual production activities.

[0003] Using exogenous strigolactone analogs to regulate the level of plant SLs to adjust the basic phenotype of plants, so that the plant type tends to be ideal, and the yield and quality of the plants are optimal, which has great economic potential. In addition, effective SLs analogs can also alleviate the adverse losses caused by environmental factors such as soil drought, soil salt stress, and abnormal high temperature climate to a certain extent. In today's climate change accelerates and extreme weather increases, it will have a great and broad application prospect. Therefore, it is of great significance to find a strigolactone analog with low use cost and good effect. SUMMARY

[0004] The present application aims at at least solving one of the technical problems existing in the prior art. To this end, the present application proposes application of pyrazoles as strigolactone analogs.

[0005] The present application also proposes a method for regulating the length of the hypocotyl of a plant.

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

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

[0008] The present application also proposes a method for regulating the plant height.

[0009] According to the first aspect of the present application, application of pyrazoles as strigolactone analogs in any of the following is proposed: (1) regulating the length of the hypocotyl of a plant; (2) promoting seed germination of a plant; (3) regulating the length and / or density of root hairs of a plant; (4) regulating the plant height.

[0010] In some embodiments of the application, the modulating hypocotyl length of the plant comprises inhibiting hypocotyl elongation of the plant.

[0011] In some embodiments of the application, the modulating root hair length and / or density of the plant comprises increasing root hair length and / or density of the plant.

[0012] In some embodiments of the application, the modulating plant height comprises inhibiting or increasing plant height.

[0013] In some embodiments of the application, the plant comprises a Brassicaceae plant.

[0014] In some embodiments of the application, the Brassicaceae plant comprises Arabidopsis thaliana.

[0015] According to a second aspect of the present application, there is provided a method of modulating hypocotyl length of a plant, the method comprising the steps of: sowing a seed of the plant in a culture reagent containing pyrazoles, and culturing.

[0016] In some embodiments of the application, the concentration of pyrazoles 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, 0.1 µM.

[0017] In some embodiments of the application, the step of culturing comprises vernalization.

[0018] In some embodiments of the application, the plant comprises a Brassicaceae plant.

[0019] In some embodiments of the application, the Brassicaceae plant comprises Arabidopsis thaliana.

[0020] According to a third aspect of the present application, there is provided a method of promoting germination of a seed of a plant, the method comprising the steps of: placing the vernalized seed in a container containing a solution of pyrazoles, and germinating the seed.

[0021] In some embodiments of the application, the concentration of pyrazoles in the solution is 0.01-0.05 µM. For example, it can be 0.01 µM, 0.02 µM, 0.03 µM, 0.04 µM, 0.05 µM.

[0022] In some embodiments of the application, the container comprises a petri dish.

[0023] In some embodiments of the application, the petri dish further comprises filter paper.

[0024] In some embodiments of the application, the plant comprises a Brassicaceae plant.

[0025] In some embodiments of the application, the Brassicaceae plant comprises Arabidopsis thaliana.

[0026] According to a fourth aspect of the present application, there is provided a method of modulating root hair length and / or density in a plant, the method comprising the steps of: sowing a seed of the plant in a culture reagent comprising pyrazolate, and culturing.

[0027] In some embodiments of the application, the concentration of pyrazolate 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, 0.1 µM.

[0028] In some embodiments of the application, the step of culturing comprises vernalization.

[0029] In some embodiments of the application, the step of culturing further comprises, after vernalization, culturing at 20-25 °C under conditions of 15-17 h light and 6-8 h darkness.

[0030] In some embodiments of the application, the step of culturing further comprises, after vernalization, culturing at 22 °C under conditions of 16 h light and 8 h darkness.

[0031] In some embodiments of the application, the plant comprises a Brassicaceae plant.

[0032] In some embodiments of the application, the Brassicaceae plant comprises Arabidopsis thaliana.

[0033] According to a fifth aspect of the present application, there is provided a method of modulating plant height, the method comprising the step of: applying a solution comprising pyrazolate to a plant.

[0034] In some embodiments of the application, the concentration of pyrazolate in the solution is 1-25 µM.

[0035] In some embodiments of the application, the concentration of pyrazolate 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, 20 µM.

[0036] In some embodiments of the present application, when the plant height is reduced, the concentration of the pyrazolate 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.

[0037] In some embodiments of the present application, the amount of the solution applied is 0.1-3 mL / each / plant.

[0038] In some embodiments of the present application, the amount of the solution applied is 1-3 mL / each / plant.

[0039] In some embodiments of the present application, the amount of the solution applied is 2 mL / each / plant.

[0040] In some embodiments of the present application, the number of times of application of the solution is 2-4 times.

[0041] In some embodiments of the present application, the number of times of application of the solution is 3 times.

[0042] In some embodiments of the present application, the period of application to the plant is after the plant bolts.

[0043] In some embodiments of the present application, the plant includes a cruciferous plant.

[0044] In some embodiments of the present application, the cruciferous plant includes Arabidopsis thaliana.

[0045] According to some embodiments of the present application, at least the following beneficial effects are achieved: the pyrazolate in the present application can be used as a strigolactone analogue to regulate the length of the hypocotyl, the seed germination, the length and / or density of the root hair, and the plant height, and has similar effects as the strigolactone analogue rac-GR24; and the pyrazolate is inexpensive, can be applied to various plants, has little environmental pollution, and can be applied on a large scale in agricultural production.

[0046] The specific agricultural production applications that can be expected are as follows: the pyrazolate provided in the present application has an inhibitory effect on the hypocotyl, and relevant research shows that the inhibition of the hypocotyl can accelerate the establishment of the light form of the seedling, save energy on the hypocotyl for the growth of the root system and young leaves, and in some cases, increase the survival rate of the seedling; the regulation of the plant height is a key topic in agricultural production and research, and further affects the yield, quality, and other factors of the crop. BRIEF DESCRIPTION OF DRAWINGS

[0047] Figure 1 The structural formula of the pyrazolate in the embodiments of the present application and the molecular docking analysis result graph; wherein A is the structural formula of the pyrazolate molecule, and B is the molecular docking result graph of the pyrazolate; Figure 2 The figure of the influence of pyrazolate on the germination of Arabidopsis thaliana in the embodiment of the present application; wherein A is the figure of the detection result of the change of the germination rate of Arabidopsis thaliana, and B is the figure of the result of the change of the germination potential of Arabidopsis thaliana; Figure 3 The figure of the influence of pyrazolate on the length of hypocotyl of Arabidopsis thaliana in the embodiment of the present application, wherein “**” represents p<0.01; Figure 4 The figure of the influence of pyrazolate on the root hair growth of Arabidopsis thaliana in the embodiment of the present application, wherein A is WT Arabidopsis thaliana, B is max4 mutant Arabidopsis thaliana, and C is d14 mutant Arabidopsis thaliana; the scale length in the figure is 1 μm. DETAILED DESCRIPTION

[0048] The concept and the technical effects of the present application will be described in detail in combination with the embodiments, so as to fully understand the purposes, features and effects of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, but not all the embodiments. Based on the embodiments of the present application, other embodiments obtained by those skilled in the art without creative labor are within the protection scope of the present application. The test methods used in the embodiments are conventional methods unless otherwise specified. The materials, reagents and the like used in the embodiments are commercially available unless otherwise specified.

[0049] Reagents and materials: Preparation of pyrazolate (purchased from Bailingwei Technology Co., Ltd.) stock solution: using a thousandth balance to weigh the reagent pure product corresponding to the required mass, dissolving and adding ultrapure water to 10 mL to prepare a 10 mM stock solution, taking 400 μL from the 10 mM stock solution, adding 1.6 mL of ultrapure water into a 2 mL EP tube to prepare a 2 mM stock solution; taking 40 μL of 10 mM pyrazolate stock solution and adding it into a 2 mL EP tube containing 1.96 mL of ultrapure water to prepare a 0.2 mM stock solution.

[0050] Preparation of rac-GR24 (purchased from Beijing Solabio Technology Co., Ltd.) stock solution: using a thousandth balance to weigh the reagent pure product corresponding to the required mass, dissolving and adding ultrapure water to 10 mL to prepare a 10 mM stock solution.

[0051] The relative molecular mass of the pyrazolate and rac-GR24 reagent pure product and the required mass are shown in Table 1.

[0052] Table 1

[0053] Preparation of solid MS medium: The 1 / 2 solid MS medium used in the experiment was prepared according to the proportion shown in Table 2. Analytical reagent sucrose (purchased from National Pharmaceutical Group Chemical Reagent Co., Ltd.), analytical reagent agar powder (purchased from Beijing Solaybao Technology Co., Ltd.) and MS medium base salt (purchased from Beijing Coolabo Technology Co., Ltd.) were weighed respectively. The mass of each component was reduced in proportion to the volume used. Sucrose and MS powder were dissolved first. After adjusting the pH of the mixed sucrose MS powder solution to 5.8-6.0 with 1% KOH solution, pour the solution into the agar powder, shake well, seal and wait for steam sterilization for standby.

[0054] Table 2 Proportion of each component of 1L 1 / 2 solid MS medium

[0055] The Arabidopsis thaliana in the examples was sowed and vernalized according to the following steps: 1) Wash the conical flask and rinse with pure water. Measure the required ultrapure water with a measuring cylinder and pour it into the conical flask. Weigh the required medicine with a one-thousandth scale, seal with a sealing film after dissolving and adjusting the pH, and standby.

[0056] 2) Take the required number of culture dishes, wash and dry them, and then put them in the steam sterilization pot together with the items that need to be sterilized (culture dishes, culture medium, pipette tip, ultrapure water, 0.1% agar). After sterilization, place the culture dishes, pipette tips and culture medium in a 65°C oven for drying or standby.

[0057] 3) Place the culture dishes, pipettes, pipette tips, sealing films, ultrapure water, 0.1% agar, 75% ethanol, and markers in the clean bench. After ultraviolet sterilization for 45 min, ventilate for 15 min. Light the alcohol lamp in the clean bench and start sowing.

[0058] 4) Take the culture medium out of the oven, add the corresponding pyrazole or GR24 mother liquor to the culture medium after slight cooling, mix well, pour into the culture dishes with labels on the bottom, and cool for standby.

[0059] 5) Take an appropriate amount of Arabidopsis thaliana seeds in a 2 mL EP tube, soak in 75% ethanol for 10 min, then remove the alcohol, wash with ultrapure water for 4-5 times, and then add an appropriate amount of 0.1% agar to the Arabidopsis thaliana seeds until the Arabidopsis thaliana mixture can be easily and smoothly sucked by a 1000 μL pipette. Thus, the seed treatment work is completed.

[0060] 6) Use 1000 μL pipette tip to suck up a certain amount of Arabidopsis seed mixed solution, remove the tip that has sucked up the seeds, and use the tip to evenly seed the Arabidopsis seeds one by one on the completely solidified medium. After the entire medium is seeded, place the seeded dish aside with the dish cover open to wait for the 0.1% agar water to evaporate. Continue to seed the other treated dishes in a similar manner.

[0061] 7) After the 0.1% agar water is completely evaporated, place the dish above the flame of an alcohol lamp, burn the dish opening with the flame, and then seal the dish with sealing film.

[0062] 8) After all the dishes are completely sealed and the sealing effect is checked twice, place the sealed dishes in a 4°C constant temperature refrigerator, vernalize for 2 days, and thus complete the seeding work.

[0063] Example 1 This example provides the application of pyrazolate as a strigolactone analogue, and the specific screening verification process is as follows: 1. Analysis of the structural properties of pyrazolate The molecular formula of pyrazolate is C 19 H 16 Cl2N2O4S, which was developed by San'ei Chemical Industry Co., Ltd. in the 1970s, is a kind of hydroxyphenylpyruvic acid dioxygenase inhibitor that affects the synthesis of some plant plastoquinone and tocopherol, thereby affecting the growth of some plants.

[0064] Through a large number of screening and research, the present application screens out pyrazolate, a structural analogue of SLs, from an organic molecule library. Pyrazolate and D14 protein are tested for molecular docking using AutoDock Vina, and the results are shown in Figure 1 From Figure 1 it can be seen that the binding energy of the best binding conformation in multiple docking results is -7.72 kcal / mol, and the binding energy of the best conformation in the molecular docking test of GR24 and D14 is -8.34 kcal / mol. Generally, the lower the binding energy value, the higher the binding feasibility. If the best conformation of the combination of GR24, pyrazolate and D14 is displayed in the same molecular model diagram, it can be observed that the binding sites of the two small molecules and D14 protein are very close.

[0065] Based on the above, it can be inferred that pyrazolate may bind to D14 protein, and D14 protein is an extremely important protein in the SLs signaling pathway. Pyrazolate may play a role at the SLs level by binding and interacting with D14 protein, so pyrazolate is subjected to subsequent verification.

[0066] 2. Effect of pyrazolate on seed germination of Arabidopsis Experimental scheme: Treatment of Arabidopsis seeds: Take a certain amount of WT Arabidopsis seeds, add ultrapure water to the EP tube, shake to immerse the Arabidopsis seeds, and put them in a 4°C refrigerator for 2 days. 3.5 μL of 100 μM pyrazolat mother liquor, 17.5 μL of 2 mM rac-GR24 mother liquor are dissolved in 7 mL of ultrapure water to prepare the treatment solution, i.e. the pyrazolat concentration is 0.05 μM and the rac-GR24 concentration is 5 μM. Pour the treatment solution into a culture dish lined with filter paper (the blank control group is directly poured into 7 mL of ultrapure water), and place the vernalized Arabidopsis seeds on the filter paper and make sure the seeds are completely soaked. Pave 50 seeds for each treatment. After marking, place in a 24 h dark, 22°C condition seedling room.

[0067] Calculation of germination rate and germination potential: Use a camera to take pictures of the seeds, count the number of germinated seeds from the photos, and calculate the germination rate. The photographing time is set to 0 h, 24 h, 29 h, 34 h, and 37 h after the Arabidopsis is placed in the seedling room. After completing the experiment, the data is summarized and plotted for analysis, and the experiment is repeated 3 times.

[0068] Germination rate / % = (number of germinated Arabidopsis seeds / total number of normally germinated Arabidopsis seeds) x 100%.

[0069] Germination potential / % = (number of Arabidopsis germinated seeds in a time period / total number of normally germinated Arabidopsis seeds) x 100%.

[0070] As shown in the following Figure 2 It can be seen from Figure A that after 24 h and 29 h of treatment of Arabidopsis seeds, the germination rates of pyrazolat and rac-GR24 treatment groups are both greater than that of the blank control group. The germination rate of the pyrazolat treatment group is 41.5% at 29 h, which is 6.3% greater than the blank control group of 35.2%, and the promotion effect on germination rate is slightly smaller than that of rac-GR24. When the time comes to 34 h, the germination rates of the blank control group and the two treatment groups are similar, and at 37 h, the germination rate of the blank control group exceeds that of the two treatment groups; As can be seen from Figure B, in terms of germination potential, the initial stage of seed germination (0-29 h), the germination potential of Arabidopsis treated with pyrazolat and rac-GR24 is greater than that of the blank control group; when the time comes to 34 h, the germination rate of the blank control group is greater than that of the two treatment groups; The results show that treatment with rac-GR24 and pyrazolat can promote the early germination of Arabidopsis seeds, which is similar to the results of using GR24 to treat cress seeds in related technologies.

[0071] 0.05 μΜ pyrazolate treatment can promote the early germination of Arabidopsis seeds, and the 24 h~29 h seed germination potential after treatment is similar to that of rac-GR24. This shows that 0.05 μΜ pyrazolate also has the function of breaking seed dormancy.

[0072] 3. Effect of pyrazolate on hypocotyl length of Arabidopsis Experimental method: Four sets of 150 mm x 25 mm sterilized glass culture dishes and four 60 mL 1 / 2 solid MS medium were prepared; the medium for cultivating Arabidopsis seedlings was set up with one control group and three experimental groups containing different concentrations of pyrazolate. The concentrations of pyrazolate in the experimental groups were 0.01, 0.03, and 0.05 μΜ, respectively. 60 μL, 180 μL, and 300 μL of 10 mM pyrazolate stock solution were added to the 1 / 2 solid MS medium to prepare the experimental medium. The control group was poured into the culture dish with 1 / 2 solid MS medium without adding reagents. WT and max4 The four types of mutant Arabidopsis were point seeded and vernalized, and then the culture dishes were transferred to 22°C room temperature and cultured under weak light conditions of 16 h light and 8 h darkness.

[0073] After 7 days of Arabidopsis culture, the hypocotyl length was measured. The normally grown Arabidopsis seedlings on the medium were selected, and according to the different treatment methods and types, they were evenly and parallelly placed on the black agar background plate with a 1 cm length reference system. A Canon EOS 5DsR camera (the camera used below is of this model) was used to take pictures of the Arabidopsis seedlings on the plate, and a MACRO LENS EF 100 mm f / 2.8L IS USM macro lens (if not specified below, it is used with this lens) was used. During the shooting process, a tripod was used to ensure that the camera focal plane was as parallel as possible to the plane where the Arabidopsis seedlings were located. After shooting, the path measurement function of the software Digmizer 6 was used to measure the hypocotyl length of the Arabidopsis seedlings one by one, and the experiment was repeated 3 times.

[0074] The results are shown in Figure 3 As shown in the results, the hypocotyl length of WT Arabidopsis treated with pyrazolate was inhibited compared with the blank control group without treatment, and the hypocotyl length was significantly shortened, but the inhibition of WT Arabidopsis hypocotyl length by pyrazolate did not show a concentration-dependent effect within the concentration gradient set in the experiment.

[0075] Observation max4 The results of the experimental group and the control group of mutant Arabidopsis showed that max4 The hypocotyl of the control group of mutant Arabidopsis was significantly longer than that of the wild type control group, and after adding pyrazolate for treatment, max4The length of hypocotyls of mutants was shortened. max4 The hypocotyl length of the mutant experimental group was slightly smaller than that of the WT control group. max4 The inhibitory effect on the hypocotyl length of mutant Arabidopsis thaliana did not show concentration dependence within the concentration gradient set in the experiment.

[0076] Further analysis of the above experimental results showed that the inhibition of pyrazolamide on WT hypocotyls was similar to the results of the previous treatment of WT Arabidopsis with rac-GR24, which indicated that pyrazolamide may have similar functions to SLs. max4 The experimental results of mutants also provide a basis for this speculation. max4 The mutant itself cannot synthesize SLs normally, but the signal transduction pathway is normal. If exogenous SLs are added, the Arabidopsis thaliana can restore the WT phenotype to a certain extent. The results presented in the experiment are exactly the same. After adding pyrazolidine treatment, max4 The hypocotyls of the WT mice were retracted to a length similar to that of the WT mice.

[0077] 4. Effects of pyrazol on root growth of Arabidopsis seedlings Experimental method: Prepare three sets of 90 mm × 15 mm high-transparency quartz glass culture dishes and three 30 mL 1 / 2 solid MS culture medium. Set up a blank control group, a pyrazolamide-treated experimental group, and a rac-GR24 positive control group. The pyrazolamide concentration in the experimental group culture medium was 0.05 μM, and the SLs analog rac-GR24 concentration used in the positive control was 5 μM. Pipette 7.5 μL of 0.2mM stock solution of pyrazolamide and 75 μL of 2 mM stock solution of rac-GR24 reagent and add them to 1 / 2 solid MS culture medium to prepare the experimental group and positive control group culture medium respectively; for the blank control, pour 1 / 2 solid MS culture medium without adding reagents onto the plate. d14 、 max4 The three mutant Arabidopsis plants were seeded and vernalized, and then the culture plates were transferred to room temperature at 22°C and cultured under 16 h light and 8 h dark conditions.

[0078] After 10 days of culture, observe the roots of the Arabidopsis seedlings. Carefully remove the seedlings from the culture medium, ensuring the integrity of the root system as much as possible. Use clean water to clean any remaining agar from the roots. Place the seedlings on a black background plate (the black background plate is an agar plate with black ink added to ensure the roots remain moist). Observe the root growth of each Arabidopsis plant and select representative Arabidopsis plants. Spread the roots of these selected plants as much as possible until there is no overlap.

[0079] Observation of root hair growth: The roots of Arabidopsis seedlings were made into temporary smears and observed under a general optical microscope. The differences in root hair in the root tip maturation zone between different treatments were observed and photographed. The experiment was repeated three times.

[0080] The root hair growth is shown in Figure 4 Fig. 3. Pyrazolate promoted the number and length of root hairs in WT and d14 , max4 . The number of root hairs in the root tip maturation zone of the three Arabidopsis plants treated with 0.05 μM pyrazolate was increased compared with the blank control group without treatment, and the length of the root hairs was also slightly longer than that of the blank control group. However, whether the increase in the number of root hairs or the elongation of the root hairs, the effect of pyrazolate treatment was lower than that of 5 μM rac-GR24 treatment.

[0081] The results showed that 0.05 μM pyrazolate and 5 μM rac-GR24 could increase the number and length of root hairs, and the effect of rac-GR24 was more significant. The results showed that pyrazolate might have a similar effect as rac-GR24.

[0082] 5, Effect of pyrazolate on the height of Arabidopsis Experimental method: Culture of Arabidopsis: Four types of Arabidopsis seeds, WT and d14 , max2 , max4 mutant, were directly sown in untreated 1 / 2 solid MS medium. After vernalization, the medium was transferred to a seedling room (22°C room temperature, 16 h light and 8 h darkness) for growth. When the Arabidopsis grew to 4-6 leaves (about 7 days later), the Arabidopsis seedlings were transplanted into small pots of 10 cm x 10 cm to continue growing, with 5 plants per small pot.

[0083] Preparation of treatment solution: Pyrazolate and rac-GR24 were used to treat Arabidopsis in the experiment. The concentrations of pyrazolate treatment solution were 5, 10, and 20 μM, and the concentration of GR24 treatment solution was 10 μM. Pyrazolate 2 mM stock solution was taken 40, 80, and 160 μL and diluted with deionized water to 16 mL to prepare pyrazolate treatment solutions of 5, 10, and 20 μM, respectively. Rac-GR24 stock solution was taken 80 μL and diluted with deionized water to 16 mL to prepare a 10 μM GR24 treatment solution.

[0084] Treatment of Arabidopsis materials: After the Arabidopsis was transplanted into pots and continued to grow for 15 days, it began to bolt. Arabidopsis with basically the same growth vigor was selected, and four Arabidopsis plants were set as a group (one pot). WT and max2 mutant plants were treated with deionized water as a control group.d14 and max4 Two mutants were used as experimental groups, and each group was treated with pyrazole and rac-GR24. 1000 μL pipette was used to drop the treatment liquid from the center of the Arabidopsis plant, 2 mL of treatment liquid was dropped on each Arabidopsis plant, and a mark was made. On the 3rd day and the 8th day after the first drop, the same method was used to apply the second and third treatments to each type of Arabidopsis. After the treatment, the four types of Arabidopsis were cultured at 22°C, 16 hours of light and 8 hours of darkness in a greenhouse, and the relative humidity of the nutrient soil was maintained. If watering is needed, it is added in a small amount from the four corners of the culture pot. 10 days after the first treatment, the height of the Arabidopsis was measured and the average value was taken.

[0085] Table 3 Height of each mutant Arabidopsis

[0086] Under the current experimental design, the results of using pyrazole to treat Arabidopsis are shown in Table 3. From the table, it can be seen that after the treatment of 5, 10, and 20 μM pyrazole on the mutant, the height corresponding to the first two concentrations is similar and slightly smaller than the blank control group, and the height of the 20 μM treatment is increased by about 20.6%. In the positive control, the height after 10 μM rac-GR24 treatment is increased by about 20.6%. max4 After the treatment liquid was applied to the mutant, the height was reduced to varying degrees. The height reduction after treatment with 10 μM rac-GR24, 5 μM pyrazole, 10 μM pyrazole, and 20 μM pyrazole was 5.1%, 18.6%, 8.7%, and 2.8%, respectively.

[0087] d14

[0088] In summary, the treatment results of pyrazole on each type of Arabidopsis are highly similar to the treatment results of the rac-GR24 positive control group, indicating that pyrazole also belongs to the analog of SLs.

[0089] The embodiments of the present application are described in detail above, but the present application is not limited to the above embodiments, and various changes can be made within the knowledge of those skilled in the art without departing from the spirit of the present application. In addition, the embodiments of the present application and the features in the embodiments can be combined with each other without conflict.​​

Claims

1. Use of pyrazolamide as a strigolactone analogue in any of the following: (1) Regulate the length of plant hypocotyls; (2) Promote plant seed germination; (3) Regulating the length and / or density of plant root hairs; (4) Regulating plant height; The plant is Arabidopsis thaliana.

2. The use according to claim 1, characterized in that The regulating plant hypocotyl length comprises inhibiting plant hypocotyl elongation; And / or, said regulating the length and / or density of plant root hairs comprises increasing the length and / or density of plant root hairs; And / or, regulating plant height includes inhibiting or increasing plant height.

3. A method for regulating plant hypocotyl length, characterized in that: The method comprises the following steps: planting plant seeds in a culture agent containing pyrazolamide for culturing; the concentration of pyrazolamide in the culture agent is 0.01-0.1µM; The plant is Arabidopsis thaliana.

4. A method for promoting plant seed germination, characterized in that: The method comprises the following steps: placing the vernalized seeds in a container containing a pyrazolamide solution for germination culture; the concentration of the pyrazolamide solution is 0.01-0.05 μM; The plant is Arabidopsis thaliana.

5. A method for regulating the length and / or density of plant root hairs, characterized in that: The method comprises the following steps: planting plant seeds in a culture agent containing pyrazolamide for culturing; the concentration of pyrazolamide in the culture agent is 0.01-0.1µM; The plant is Arabidopsis thaliana.

6. A method for regulating plant height, characterized in that: The method comprises the following steps: applying a solution containing pyrazolamide to a plant; the effective concentration of pyrazolamide in the solution is 1-25 μM; The plant is Arabidopsis thaliana.

7. The method according to claim 6, characterized in that The application amount of the solution is 0.1-3 mL / time / plant.

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