Use of AZD8055 compound in improving seed vigor of crops

Abstract

The present invention relates to the technical field of regulating the seed vigor of crops, and in particular to a use of an AZD8055 compound in improving the seed vigor of crops. The AZD8055 compound has the following chemical structure: formula (I). The AZD8055 compound can effectively improve seed vigor, and promote rapid seed germination, thereby leading to robust and uniform seedling emergence.

Description

Uses of compound AZD8055 in improving crop seed vigor Technical Field

[0001] This invention relates to the field of crop seed vigor regulation technology, specifically to the use of compound AZD8055 in improving crop seed vigor. Background Technology

[0002] Seed vigor is a core indicator for measuring seed quality and efficacy, and it has a profound impact on the high-quality development of modern agricultural production. The level of seed vigor directly affects crop yield and quality, and also determines germination rate, seedling vigor, and subsequent production potential. High-quality, high-vigor seeds exhibit excellent storage tolerance and can germinate rapidly and uniformly under suitable conditions, forming a consistent seedling emergence pattern. This is crucial for the successful implementation of direct-seeding methods for crops such as rice and corn, and also plays a decisive role in optimizing field planting density, enhancing the competitive advantage between crops and weeds, and improving the ability to resist pests, diseases, and abiotic stresses. International data shows that approximately one-fifth of the global seed market suffers significant economic losses due to challenges such as uneven germination, weak stress response, and poor seedling growth caused by seed vigor issues.

[0003] Therefore, the research and application of seed vigor enhancement technologies have become crucial for ensuring national food security, promoting agricultural modernization, improving the economic benefits of simplified planting methods such as direct seeding, and achieving goals such as increased yield per unit area and reduced inventory costs. The transformation process from seed dormancy to germination and seedling emergence is the core step reflecting its vigor attributes and a landmark event marking the beginning of the plant's life cycle, holding an irreplaceable standard for judging seed quality. In my country, a 1% increase in corn seed germination rate theoretically increases yield by approximately 36 jin per mu (0.067 hectares). If this improvement can be widely applied nationwide, the total corn yield is expected to increase dramatically to a staggering 21.8 billion jin (14.9 million tons), generating enormous economic and social benefits.

[0004] Research has revealed that seed dormancy and germination are complex biological phenomena involving gene expression regulation, environmental signal sensing, and the synergistic effects of multiple biochemical pathways. Among these, the plant hormone abscisic acid (ABA) plays a crucial role in seed vigor regulation. ABA, an important hormone widely present in vascular plants, participates in the entire life cycle of seeds from development to maturity, including but not limited to dormancy induction, germination initiation, stomatal movement regulation, fruit ripening control, and various stress responses. Taking rice as an example, transcription factors bZIP23 and bZIP42 actively regulate seed vigor by directly binding to and activating the transcriptional activity of the Peroxiredoxin 1A (PER1A) promoter region. Changes in the expression levels of these two factors are directly affected by ABA concentration, indicating that the ABA signaling pathway may finely adjust seed vigor through the bZIP23-PER1A signaling axis. During seed maturity, the endogenous concentration of ABA rises to its peak, finely regulating the accumulation of reserve substances such as lipids, starch, and storage proteins to prevent premature germination and maintain seed dormancy. The latest research also revealed the mechanism by which OsUGT75A accelerates seed germination by reducing ABA content. Correspondingly, stored seeds generally accumulate higher ABA concentrations, which inhibit seed germination and seedling emergence through their signaling flux.

[0005] Although some substantial progress has been made in the study of the degradation mechanism of regulatory factors in the ABA signaling pathway, such as the revelation in the model plant Arabidopsis thaliana that the ubiquitination system is involved in the degradation of ABA pathway regulatory factors ABA Insensitive 3 (ABI3) and ABI5 proteins, current research focuses more on the proteasome degradation mechanism mediated by the ubiquitination pathway. Moreover, most of this work is related to the ABA-mediated plant response to adverse conditions such as drought, and no studies have been found on using reagents to regulate ABA to enhance seed vigor. Summary of the Invention

[0006] The purpose of this invention is to overcome the shortcomings of the prior art and provide the use of the AZD8055 compound in improving crop seed vigor. The AZD8055 compound can effectively enhance seed vigor, promote rapid seed germination, and then result in robust and uniform seedling emergence.

[0007] To achieve the above objectives, the present invention provides the following technical solution:

[0008] Provided is the use of the compound AZD8055 in improving crop seed vigor, said compound AZD8055 comprising the chemical structure shown in general formula (I):

[0009] in,

[0010] R 1 Selected from CH2X, among which,

[0011] X is selected from OH. NHCH2CH2OH, NHCH2CH2OMe, N(CH2CH2OH)2, NHCH2CH2NHMe, NHCH2CH2NMe2, NHCO(OR), NHCH2CO2Et, NHCH2CO2H, NHMe'NMe2, or R 1 Selected from CONH2, CONHoH, CONHMe, CONHCH2CH2OH, CONHCH2CH2OMe, CONHCH2CH2NHMe, CSNH2.

[0012] In some embodiments, the AZD8055 compound is any one of the following chemical structures:

[0013] The use of the aforementioned AZD8055 compound in the preparation of drugs that enhance crop seed vigor is also provided.

[0014] It also provides a method to improve crop seed vigor by using the aforementioned AZD8055 compound to target the seed ABA signaling pathway to improve crop seed vigor.

[0015] A method for synthesizing the compound represented by the above general formula (I) is also provided, characterized by comprising the following steps:

[0016] 2,6-dichloro-6-carboxypyridine synthesis

[0017] Will synthesis

[0018] Will Chemical structure of general formula (I)

[0019] The beneficial effects of using the AZD8055 compound of this invention in improving crop seed vigor:

[0020] This invention discovers that pyrido[2,3-D]pyrimidine and 4-methoxy-phenyl in compound AZD8055 can regulate the ABA signaling pathway, thereby effectively enhancing the seed vigor of various crops, promoting rapid seed germination and robust and uniform seedling emergence. This addresses the issue of decreased seed vigor due to seed storage, facilitates successful direct seeding, and significantly boosts crop yield per acre, resulting in substantial social benefits. This invention is suitable for large-scale production and application. Attached Figure Description

[0021] Figure 1 shows the effect of different concentrations of treatment on the germination rate of the rice variety Zhonghua 11 in Experiment 1.

[0022] Figure 2 shows the effect of treatment 1 on the seed vigor of different japonica rice varieties.

[0023] Figure 3 shows the effect of treatment 1 on the seed vigor of different indica rice varieties.

[0024] Figure 4 shows the germination rate of different crop seeds after treatment with AZD8055 in Experiment Example 2.

[0025] Figure 5 shows the germination rate of crop seeds after treatment at different storage periods in Experiment Example 3. Detailed Implementation

[0026] Preferred embodiments of the invention will now be described in more detail with reference to the accompanying drawings. While preferred embodiments of the invention are shown in the drawings, it should be understood that the invention can be implemented in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided so that the invention will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.

[0027] Example 1

[0028] To illustrate the method for obtaining the AZD8055 compound of this invention, the following is disclosed:

[0029] As shown in Figure 6, the basic skeleton of the AZD8055 compound is composed of basic skeleton 1 (pyrido[2,3-D]pyrimidine) and basic skeleton 2 (4-methoxy-phenyl), which are the basic structures for achieving mTOR inhibition activity and thus regulating seed vigor. On this basic skeleton, the 3-position derivatization of the benzene ring of basic skeleton 2 and the 2- and 4-position derivatization of basic skeleton 1 can better realize the effect of the AZD8055 compound in improving seed vigor.

[0030] It was found that introducing a hydrophilic group into the R1 group can improve IC. 50 Results. To reduce the difficulty of synthesis, R2 and R3 can be designed with the same structure, or they can be designed with different structural fragments.

[0031] This yields the following derived groups:

[0032] Note: The structure within the dashed box should be given priority.

[0033] The symbol “=" above means selected from.

[0034] To illustrate how the basic framework of the AZD8055 compound was synthesized, the synthetic approach is explained below:

[0035] Based on the retrosynthetic method, the analysis is as follows:

[0036] Therefore, the method for synthesizing the basic skeleton of the compound AZD8055 shown in general formula (I) includes the following steps:

[0037] 2,6-dichloro-6-carboxypyridine synthesis

[0038] Will synthesis

[0039] Will Chemical structure of general formula (I)

[0040] Those skilled in the art can implement the specific synthesis steps through experimental methods.

[0041] Specifically, compound AZD8055 with the following designation was obtained, where the designation is for labeling purposes only:

[0042] Effect verification:

[0043] To illustrate the effect of the AZD8055 compound of the present invention in enhancing seed vigor, the following experiments were conducted, wherein the AZD8055 compound described in the experimental examples is exemplarily:

[0044] Drug Name: AZD-8055

[0045] CAS No.: 1009298-09-2

[0046] Molecular formula: C 25 H 31 N5O4

[0047] Molecular weight: 465.54

[0048] Chemical structure:

[0049] The application of compound AZD8055 to enhance seed vigor in different crops is detailed below:

[0050] 1.1 Seed germination experiment: The method was slightly modified from Wang et al. (2010). Seeds of popular varieties of rice, sweet corn, soybean, tomato, cucumber, water spinach, and Chinese cabbage, along with their remaining stock, were used as the test subjects. Three replicates were set up for each material. For each replicate, 50 mature and plump seeds were selected and placed in 9cm diameter transparent disposable petri dishes lined with filter paper. 10mL of distilled water was added, and the petri dishes were then placed in a 28℃ constant temperature and light incubator (12h light / 12h darkness) for 5 or 7 days. Germination was defined as the embryo breaking through the seed coat by 2mm, and seedling establishment was defined as the radicle reaching the length of the seed and the embryo reaching more than half the length of the seed. The number of germinated and established seeds was counted every 12 hours. The germination rate over 5 consecutive days is taken as the germination potential (GP) over 5 days. Germination index (GI) = ∑(Gt / Dt) (Dt represents the number of germination days), Gt represents the number of seeds germinated each day corresponding to Dt, and T50 is the time (d: days) required for the germination rate to reach half.

[0051] 1.2 Germination Experiment with Exogenous Reagent Treatment: For different crops, appropriate concentrations of reagents were selected for external application. The germination rate was recorded 3-5 days after application under different concentrations, and the germination potential was recorded for 5 consecutive days.

[0052] 1.3 Seedling growth assessment experiment:

[0053] (1) Select 50 mature and plump seeds, gently peel off the husk, and try to avoid damaging the embryo.

[0054] (2) Place the shelled seeds in a 50℃ oven for about 3 days to break dormancy.

[0055] (3) Then pour the seeds into a 50mL sterile centrifuge tube, add 10mL of 75% anhydrous ethanol, wash the seeds with a shaker at 200rpm for 5min, pour out the ethanol in a clean bench (be careful not to drop the seeds), and wash once with sterile water.

[0056] (4) Add 5 mL of 2.5% sodium hypochlorite to the laminar flow hood and wash the seeds with a shaker at 200 rpm for 15-20 min. Then discard the sodium hypochlorite in the laminar flow hood and wash with sterile water 3-5 times.

[0057] (5) In a clean bench, place the washed seeds on sterile filter paper to absorb the moisture. Use tweezers that have been sterilized by high temperature after flaming to carefully pick up the dried seeds and place them on a 1 / 2 MS medium plate. After inoculation, seal the culture dish with sealing film and place it in a dark incubator at 28℃ for 2 days.

[0058] (6) After most of the seeds show white sprouts, disinfect the surface with alcohol and open it in a clean bench. Inoculate the seeds with consistent germination onto 1 / 2MS medium plates containing different concentrations of reagents. After sealing, place them in a 28℃ constant temperature light incubator (12h light / 12h dark) for 12 days.

[0059] (7) Count the length, root length and number of rice seedlings, and take photos.

[0060] 1.4 Seedling growth experiment in soil cultivation:

[0061] Each material was prepared in triplicate. For each replicate, 30 mature, plump seeds were selected and placed in 9cm diameter transparent disposable petri dishes lined with filter paper. 10mL of distilled water or reagent treatment solution was added, and the petri dishes were then placed in a 28℃ constant temperature and light incubator (12h light / 12h dark) for 3 days. Subsequently, the seeds were transferred to pots filled with moist field soil and placed in a 28℃ constant temperature and light incubator (12h light / 12h dark) for 14 days. Seedling growth was observed and photographed during this period.

[0062] Experimental Example 1

[0063] Screening of AZD8055 compound treatment concentrations and testing on different rice varieties:

[0064] Taking the japonica rice variety Zhonghua 11 as an example, the study investigated the effect of AZD8055 on seed vigor and screened for suitable concentrations (Treatment indicates treatment). The AZD8055 compound was applied to the seeds of Zhonghua 11, and the seeds were then placed in petri dishes to await germination. Figure 1 shows that AZD8055 at concentrations of 0.5 μM, 1 μM, 2 μM, and 5 μM indeed improved seed vigor (germination rate), with the 2 μM concentration showing the best effect.

[0065] Based on the above results, other different japonica rice varieties (3: EBGopher; 10: AoChiu2Hao; 12: Bombilla; 14: BERLIN) were treated with the same concentration of 2 μM. The results in Figure 2 show that the treated materials showed significant improvements in both germination rate and seedling growth (seedling length).

[0066] To further determine the effect of AZD8055 on improving seed vigor in different rice varieties, different indica rice varieties (6126: Shengyou 6126; 51: Nayou 51; 1179: Ruanhuayou 1179; 6388: Nayou 6388) were treated with the same concentration of 2 μM. The results in Figure 3 show that all of them can significantly improve the germination rate.

[0067] It is evident that the AZD8055 compound of the present invention can achieve a significant effect in enhancing seed vigor.

[0068] Experimental Example 2

[0069] Tests on the effects of AZD8055 compound treatment on different crop seeds

[0070] To further explore the application scope of AZD8055 compound in seed vigor regulation, relevant tests were conducted on different crop varieties (wheat Kenong 199, corn, soybean Dongnong 47, tomato AC, Chinese cabbage variety Xukefengkang 80, improved red cabbage variety Shiwang, and water spinach variety Xukejianye water spinach).

[0071] The study investigated the effects of 2 μM AZD8055 on the seed vigor of wheat Kenong 199, corn, soybean Dongnong 47, and tomato AC. It was found that 2 μM AZD8055 effectively enhanced the seed vigor of all four crops.

[0072] The seeds of Chinese cabbage, red mustard greens, and water spinach were treated with 0.5 μM AZD8055. It was observed that 0.5 μM AZD8055 significantly improved the seed vigor of all three varieties. The remaining treatment steps were the same as in Experiment 1. Figure 4 shows that the germination rate of the treated seeds was significantly higher than that of the untreated seeds.

[0073] Experimental Example 3

[0074] Tests of AZD8055 compound treatment on crop seeds at different storage periods

[0075] To further explore the application scope of AZD8055 in seed vigor, treatments were performed on crop varieties at different storage periods, including soybean variety Yangchun and sweet corn varieties Huameitian 16 and Huameitian 8. Figure 5 shows that AZD8055 treatment significantly improved seed germination rate in soybean varieties stored for 3, 7, and 10 years; and also promoted germination in sweet corn varieties stored for 2 and 4 years.

[0076] It is evident that the AZD8055 compound of the present invention can enhance seed vigor. Since other AZD8055 compounds contain the same skeleton as compound 1 AZD8055, and compound 1 AZD8055 enhances seed vigor, the derivative groups of other AZD8055 compounds, which are mainly N- or O-containing alkyl or cycloalkyl groups, also have the same effect of enhancing seed vigor.

[0077] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. The use of compound AZD8055 in improving the germination rate, seedling growth, and seedling height of rice seeds, wherein the chemical structural formula of compound AZD8055 is as follows: The concentrations of the AZD8055 compound were 0.5 μM, 1 μM, 2 μM, and 5 μM.

2. The use of compound AZD8055 in improving the germination rate of wheat, corn, soybean, tomato, Chinese cabbage, red mustard greens, and water spinach seeds, wherein the chemical structural formula of compound AZD8055 is as follows: The concentration of compound AZD8055, which improves the germination rate of wheat, corn, soybeans, and tomatoes, is 2 μM. The concentration of compound AZD8055, which improves the germination rate of Chinese cabbage, red cabbage, and water spinach seeds, is 0.5 μM.

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