Novel indole plant growth regulator as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of vegetation restoration, and relates to a novel plant growth regulator, in particular to a novel plant growth regulator containing an indole structure as well as a preparation method and application of the novel plant growth regulator. The plant growth regulator provided by the invention is an indole compound synthesized by taking o-phenylenediamine and indole-3-formaldehyde as raw materials, and crop growth can be promoted or inhibited under different concentration conditions by applying regulator solutions with different concentrations. And IYAA can generate complexation with copper by virtue of the structural characteristics of IYAA, so that the growth of plants under copper stress is promoted. The regulator provided by the invention is simple in preparation method, obvious in effect and wide in application prospect.

Description

Technical Field

[0001] The invention belongs to the technical field of vegetation restoration and relates to a novel plant growth regulator, in particular to a novel plant growth regulator containing an indole structure and a preparation method and application thereof. Background Art

[0002] Plant growth regulators are exogenous active substances produced by artificial synthesis, artificial extraction or biological fermentation. They have similar physiological activities to endogenous plant hormones, or can affect the synthesis, transportation, metabolism and physiological effects of endogenous plant hormones. Plant growth regulators have the functions of promoting seed germination, rooting and flowering, enhancing the ability of plants to resist cold, drought, salinity and pests, and improving crop yield and quality. They have the advantages of low cost, small dosage and significant effect, and are widely used in agricultural production.

[0003] Plant growth regulators are a major industry that has attracted much attention and development from countries around the world, and are also the main focus of the future agricultural field. The scientific use of plant growth regulators can reduce the use of pesticides and fertilizers, thereby reducing environmental pollution. It not only solves food security issues to a certain extent and meets people's growing demand for high-quality agricultural products, but also alleviates increasingly severe environmental problems to a certain extent. The development of new plant growth regulators has become a research hotspot. Summary of the invention

[0004] In view of the problems existing in the prior art, the purpose of the present invention is to provide a new type of plant growth regulator IYAA containing an indole structure for the first time. The plant growth regulator provided by the present invention can promote the growth of wheat, and the operation method is simple, the effect is good, and it has broad application prospects.

[0005] Another object of the present invention is to provide a method for preparing the novel plant growth regulator IYAA.

[0006] The present invention also provides application of the novel plant growth regulator IYAA.

[0007] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions: The present invention provides a novel plant growth regulator having a structure shown in Formula I: Formula I.

[0008] The present invention also provides a method for preparing the novel plant growth regulator, comprising the following steps: mixing o-phenylenediamine and indole-3-carboxaldehyde with an organic solvent, and performing an esterification reaction to obtain a novel plant growth regulator having a structure shown in formula I.

[0009] In some embodiments, the molar ratio of o-phenylenediamine to N-acetic acid-3-indolecarboxaldehyde is 1:1, and the solvent used is anhydrous ethanol.

[0010] In some embodiments, the esterification reaction is carried out under nitrogen protection and reflux under heating conditions.

[0011] In some embodiments, the heating temperature of the esterification reaction is 70° C. and the reaction time is 12 h.

[0012] In some embodiments, the reaction solution generated by the esterification reaction is cooled to room temperature, dried by spin drying, and then an organic solvent (ethanol) is added to cool and recrystallize, and the product is purified by filtration. The specific process of the above purification is: adding an organic solvent (ethanol) for recrystallization: first cooling until a little solid precipitates, then ultrasonically precipitating a large amount of solid, filtering to obtain a light yellow powder, repeating three times to purify the product.

[0013] The invention also provides application of the novel plant growth regulator in regulating wheat growth.

[0014] The beneficial effects of the present invention are: (1) The present invention provides a novel plant growth regulator having a structure as shown in Formula I. The plant growth regulator (IYAA) provided by the present invention is a novel plant growth regulator synthesized based on indole compounds. The compound can affect the growth of wheat seedlings by affecting the germination rate and rhizome length of wheat seeds, and in particular can achieve the same growth-promoting effect as commercial auxin, while reducing costs.

[0015] (2) The plant growth regulator provided by the present invention can react with copper ions by virtue of its structural characteristics, and can alleviate copper stress on wheat to a certain extent; (3) The plant growth regulator provided by the present invention has a simple preparation process and strong controllability. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 This is the high-resolution mass spectrum of compound IYAA; Figure 2 is the nuclear magnetic resonance (NMR) spectrum of compound IYAA; Figure 3 This is a graph showing the water solubility test results of compound IYAA; Figure 4 This is the result diagram of the effect of compound IYAA on the germination rate of wheat seeds; Figure 5 This is the result diagram of the effect of compound IYAA on the root and stem length of wheat seedlings in hydroponics; Figure 6This is the result diagram of the effect of compound IYAA on the root growth indexes of wheat seedlings cultured in soil; Figure 7 This is a graph showing the effect of compound IYAA on the root and stem length of wheat seedlings under copper stress. DETAILED DESCRIPTION

[0017] The present invention provides a method for preparing the plant growth regulator in the above technical solution, comprising the following steps: o-phenylenediamine and indole-3-carboxaldehyde were dissolved in ethanol, refluxed for 12 hours under nitrogen protection, and the reaction solution was cooled to room temperature. The liquid was spin-dried to obtain a crude product, which was recrystallized from ethanol three times and dried to obtain a light yellow powder. After drying, a novel plant growth regulator having a structure shown in Formula I was obtained.

[0018] In the present invention, unless otherwise specified, the raw materials used are commercially available products well known to those skilled in the art.

[0019] In the present invention, the molar ratio of o-phenylenediamine to indole-3-carboxaldehyde is 1:1, and ethanol is selected as the organic solvent.

[0020] The corresponding reaction is shown below.

[0021] 1 H NMR (500 MHz ， DMSO) δ 11.73 (s ， 1H) ， 8.75 (s ， 1H) ， 8.41 (d ， J = 7.5 Hz ， 1H) ， 7.99 (s ， 1H) ， 7.49 (d ， J = 7.9 Hz ， 1H) ， 7.28 – 7.16 (m ， 2H) ， 7.02 (d ， J = 7.1 Hz ， 1H) ， 6.92 (dd ， J = 10.9 ， 4.1 Hz ， 1H) ， 6.79 – 6.64 (m ， 1H) ， 6.59 (t ，J = 7.5 Hz ， 1H) ， 5.01 (s ， 2H). 13 C NMR (126 MHz ， DMSO) δ 153.17 ， 143.28 ， 138.32 ， 137.66 ， 133.57 ， 126.29 ， 125.24 ， 123.24 ， 122.32 ， 121.40 ， 117.05 ， 116.93 ， 115.95 ， 114.73 ， 112.45. HRMS (m / z): [M + H] + Calculate for C 15 H 13 N3: 236.1182 ， found ， 236.1179. In the present invention, the molar ratio of o-phenylenediamine to indole-3-carboxaldehyde is 1:1, and ethanol is selected as the organic solvent. O-phenylenediamine and indole-3-carboxaldehyde are dissolved in ethanol, and after reflux for 12 hours under nitrogen protection, the reaction temperature is 70°C. The reaction solution is cooled to room temperature, and the reaction solution is spin-dried to obtain a crude product. The crude product is recrystallized from ethanol three times, first cooled until a little solid is precipitated, and then ultrasonically precipitated a large amount of solid, filtered to obtain a light yellow powder, and repeated three times, the product is purified, and dried to obtain a light yellow powder.

[0022] The present invention provides a method for preparing the novel plant growth regulator described in the above technical solution. In the present invention, the novel plant growth regulator is used in wheat growth, for example, the novel plant growth regulator can act on different organs of wheat and affect the growth of wheat.

[0023] The technical solutions in the present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0024] Example 1 Dissolve o-phenylenediamine (1080 mg, 10 mmol) and indole-3-carboxaldehyde (1450 mg, 10 mmol) in 90 mL of ethanol. Reflux at 70 °C for 12 hours under nitrogen protection. Cool the reaction solution to room temperature, spin dry the resulting liquid, recrystallize with 10 ml of ethanol three times, and dry to obtain 900 mg of a light yellow powder (yield: 32.4%).

[0025] Figure 1 This is the high-resolution mass spectrum of compound IYAA. Figure 2 is the nuclear magnetic resonance (NMR) spectrum of IYAA. Figure 1 and Figure 2 It can be seen that the target product prepared in this example is compound IYAA.

[0026] Effect Example 1 The water solubility of the compound IYAA prepared in Example 1 was determined as follows: Compound IYAA was dissolved in pure water to obtain a concentration of 10 -3 M of IYAA stock solution; dilute to obtain a concentration of 10 -4 , 10 -5 , 10 -7 , 10 -9 , 10 -11 , 10 -13 Plant growth regulator stock solution of M.

[0027] Figure 3 The water solubility test result of compound IYAA is shown in FIG. Figure 3 It can be seen that compound IYAA has good water solubility and there is no insoluble large particles or precipitation.

[0028] Effect Example 2 The effect of the compound IYAA prepared in Example 1 on the germination rate of wheat seeds was analyzed as follows: Compound IYAA was dissolved in pure water to obtain a concentration of 10 -3 M of IYAA stock solution and diluted to a concentration of 10 -4 , 10 -5 , 10 -7 , 10 -9 , 10 -11 , 10 -13 M of plant growth regulator stock solution for later use; similarly, prepare and dilute the concentration to 10 -4 , 10 -5 , 10 -7 , 10 -9 , 10 -11 , 10 -13 The mother solution of commercial auxin IAA of M is reserved; Sodium hypochlorite solution was used to disinfect wheat seeds (5-10 min), the sodium hypochlorite solution was sucked away and clean water was added to cover the seeds, and the seeds were left standing for 30-60 min. The clean water was drained and the seeds were spread on a clean ziplock bag and placed in a ventilated place to dry. Seeds of uniform size were randomly selected and placed in glass culture dishes with filter paper laid in advance. Finally, diluted IYAA mother solution of different concentrations were added and labeled. Three mother solution treatment methods of IYAA, IAA, and CK were set, and three replicates were set for each concentration. The culture dishes were placed in a room temperature environment of 25 ℃ for hydroponics, and the same amount of mother solution was added every 24 hours.

[0029] Figure 4 The figure shows the effect of IYAA and IAA on wheat seed germination rate at different concentrations. Compared with the commercial auxin IAA, the compound IYAA showed a better promoting effect on wheat seed germination rate, especially at a concentration of 10 -7 M and 10 -9 M time.

[0030] Effect Example 3 The effect of the compound IYAA prepared in Example 1 on the root and stem growth of wheatgrass seeds under hydroponics was analyzed as follows: Dissolve compound IYAA and auxin IAA in pure water, prepare and dilute them to a concentration of 10 -4 , 10 -5 , 10 -7 , 10 -9 , 10 -11 , 10 -13 The stock solution of IYAA compound of M and commercial auxin IAA of the same concentration is ready for use; Three mother solution treatments of IYAA, IAA and CK were set up, and three replicates were set for each concentration. The culture dishes were placed in a room temperature environment of 25°C for hydroponics. An equal amount of mother solution was added every 24 hours. The culture was carried out for 7-14 days, and the root and stem length at each concentration was counted.

[0031] Figure 5 The figure shows the effect of compound IYAA and IAA on the root and stem length of wheat at different concentrations. It can be seen from the figure that compared with the commercial auxin IAA treatment group, compound IYAA showed a better promoting effect on the root and stem length of wheat seedlings, especially at medium and low concentrations (10 -7 -10 -9 M).

[0032] Effect Example 4 The experiment to analyze the effect of compound IYAA on the root and stem length of wheat seedlings cultured in soil is as follows.

[0033] Dissolve compound IYAA and auxin IAA in pure water, prepare and dilute them to a concentration of 10 -4 , 10 -5 , 10 -7 , 10 -9 , 10 -11 , 10 -13 M's IYAA compound and commercial auxin IAA are available for use; At room temperature, 10 mL of IYAA and IAA mother solutions of corresponding concentrations were used to soak the seeds for 12 hours, and a clean water control (CK) was set up at the same time. After the soaking, 52 flower pots with a diameter of 15 cm were prepared, and 50 seeds in each culture dish were randomly selected and sown into flower pots filled with appropriate amount of soil, evenly placed, and covered with 1 cm of soil moistened with clean water. Repeat four times. Every two days, a certain amount of solution (5 mL) was added uniformly with a watering can. The germination rate was counted from the second day after the seeds were planted, and continued for 5 days. Plant height was measured 5 days after planting, and 20 plants were measured for each concentration, and then measured every 3 days for 3 weeks. On this basis, after 3 weeks of continuous cultivation, root length, stem length, fresh weight, dry weight, and chlorophyll were measured.

[0034] Figure 6 The effect diagram of IYAA and IAA under different concentrations and the comparison diagram of various indicators. In the same real photo, the upper part is the effect diagram of IAA (commercial growth regulator) and CK group on wheat growth under different concentrations, and the lower part is the effect diagram of IYAA (new plant growth regulator) and CK group on wheat growth under different concentrations. From left to right, they are CK, 10 -4 , 10 -5 , 10 -7 , 10 -9 , 10 -11 , 10 -13 M group. The plant height, root length, chlorophyll, and biomass of each treatment group were counted. The bar graph on the right shows that -7 M, the growth-promoting effect of IYAA on wheat was most obvious, and it could be seen at 10 -7 IYAA also had obvious growth-promoting effects on other indicators.

[0035] Effect Example 5 The experiment to analyze the effect of compound IYAA on the root and stem length of wheat seedlings under heavy metal stress is as follows.

[0036] Dissolve compound IYAA in pure water and prepare and dilute to a concentration of 10 -4 , 10 -5 , 10 -7 , 10 -9 , 10 -11 , 10-13 M of IYAA compound for standby use; and prepare 40, 80 mg / LCu with copper sulfate pentahydrate 2+ Solution.

[0037] Set IYAA, IYAA+Cu 2+ , Cu 2+ The culture dishes were placed at room temperature of 25 ℃ for hydroponics. An equal amount of solution was added every 24 hours for 7-14 days, and the root and stem length at each concentration was counted.

[0038] Figure 7 The following are the physical pictures of the effects of different concentrations of IYAA on wheat growth under copper stress and the statistical comparison of root and stem length data. In the physical picture, the first row is the treatment group without copper stress, and the second and third rows are the treatment groups with Cu 2+ The concentrations of IYAA were 40 mg / L and 80 mg / L, respectively. From left to right in each row, the concentrations of IYAA were 0 (CK group), 10 -4 , 10 -5 , 10 -7 , 10 -9 , 10 -11 , 10 -13 M. From the actual picture and the statistical root length data, it can be found that compared with the treatment group without copper stress, under copper stress, the high concentration of IYAA did not inhibit wheat, but had the same growth-promoting effect as the lower concentration, which proved that IYAA and Cu 2+ Some kind of effect occurred, reducing the Cu 2+ concentration, thus promoting the growth of wheat.

[0039] From the above test examples, it can be seen that the novel plant growth regulator provided by the present invention can affect the root and stem length of wheat by affecting the speed of seed germination, and shows differences from the commercialized auxin IAA; the operation method is simple and has broad application prospects.

[0040] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A novel plant growth regulator, characterized in that: It has the structure shown in formula I: Formula I.

2. A method for preparing the novel plant growth regulator according to claim 1, characterized in that: The method comprises the following steps: mixing o-phenylenediamine and indole-3-carboxaldehyde with an organic solvent, and carrying out an esterification reaction to obtain a novel plant growth regulator having a structure shown in formula I.

3. The preparation method according to claim 2, characterized in that: The molar ratio of o-phenylenediamine to indole-3-carboxaldehyde is 1:

1.

4. The preparation method according to claim 2 or 3, characterized in that: The organic solvent is ethanol.

5. The preparation method according to any one of claims 2 to 4, characterized in that: The esterification reaction was carried out under nitrogen protection and reflux under heating conditions. The heating temperature was 70° C. and the reaction time was 12 h.

6. The preparation method according to any one of claims 2 to 5, characterized in that: The reaction solution generated by the esterification reaction is cooled to room temperature, spin-dried, and then an organic solvent is added to cool and recrystallize, and the product is purified by filtration.

7. The preparation method according to claim 6, characterized in that: The organic solvent is ethanol.

8. Use of the novel plant growth regulator as claimed in claim 1 in regulating wheat growth.

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