Use of AZD8055 compound in improving crop seed vigor

Abstract

The present invention relates to the technical field of regulating crop seed vigor, and specifically to a use of an AZD8055 compound in improving crop seed vigor. The AZD8055 compound comprises the following chemical structure: the AZD8055 compound can effectively enhance seed vigor, promote rapid seed germination, and subsequently achieve robust and uniform seedling emergence.

Description

Technical Field

[0001] The present invention relates to the technical field of regulating crop seed vigor, and in particular to use of the AZD8055 compound in improving crop seed vigor. Background Art

[0002] Seed vigor, a core indicator of seed quality and potency, has a profound impact on the high-quality development of modern agricultural production. Seed vigor not only directly affects crop yield and quality, but also determines post-sowing emergence, seedling vigor, and subsequent production potential. High-quality, high-vibrancy seeds exhibit excellent storability during storage, germinating quickly and evenly under favorable conditions, resulting in uniform seedling emergence. This is crucial for the successful implementation of direct-seeding planting methods for crops such as rice and corn. It also plays a decisive role in optimizing field planting density, enhancing crop-weakness competition, and improving resilience to pest and disease invasions and environmental stress. International data shows that approximately one-fifth of the global seed market suffers significant economic losses due to challenges such as uneven emergence, poor response to environmental stress, and poor seedling growth caused by seed vigor issues.

[0003] Given this, the research and application of seed vigor enhancement technologies has become crucial for ensuring national food security, promoting agricultural modernization, improving the economic benefits of simplified planting methods like direct seeding, and achieving goals such as increased yields and reduced inventory costs. The transformation of seeds from dormancy to germination is a key step in reflecting their vigor and marks the beginning of a plant's life cycle. It serves as an irreplaceable standard for evaluating seed quality. In my country, a 1% increase in corn seed emergence rate theoretically increases yield by approximately 36 kilograms per mu. If this improvement were widely implemented nationwide, total corn production is projected to surge to 21.8 billion kilograms, generating significant economic and social benefits.

[0004] Research has found that the process of seed dormancy and germination is a complex biological phenomenon, involving gene expression regulation, environmental signal sensing, and the coordinated interaction of multiple biochemical pathways. Among these, the plant hormone abscisic acid (ABA) plays a crucial role in regulating seed vigor. As a key hormone widely present in vascular plants, ABA is involved in the entire seed life cycle from development to maturity, including physiological processes such as dormancy induction, germination initiation, stomatal movement regulation, fruit ripening control, and various stress responses. In rice, for example, the transcription factors bZIP23 and bZIP42 actively regulate seed vigor by directly binding to the promoter region of Peroxiredoxin 1A (PER1A) and activating its transcriptional activity. Changes in their expression are directly affected by ABA concentration, suggesting that the ABA signaling pathway may fine-tune seed vigor through the bZIP23-PER1A signaling axis. During seed maturity, endogenous ABA concentrations rise to a peak, preventing premature germination and maintaining dormancy by finely regulating the accumulation of reserve substances such as fat, starch, and storage proteins. The latest research also revealed that OsUGT75A accelerates seed germination by reducing ABA content. Accordingly, stored seeds generally accumulate higher ABA concentrations, which inhibit seed germination through its 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, for example, in the model plant Arabidopsis, the ubiquitination system was revealed to be involved in the degradation process of the ABA pathway regulatory factors ABAInsensitive 3 (ABI3) and ABI5 proteins, more focus is currently 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, but there is no evidence of regulating ABA through reagents to enhance seed vigor. Summary of the Invention

[0006] The object of the present invention is to avoid the deficiencies in the prior art and to provide a use of the AZD8055 compound in improving the vigor of crop seeds. The AZD8055 compound can effectively enhance seed vigor, promote rapid seed germination, and subsequently produce robust and uniform seedlings.

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

[0008] Provided is the use of the AZD8055 compound in improving crop seed vigor. The AZD8055 compound comprises a chemical structure represented by general formula (I):

[0009]

[0010] in,

[0011] R 2 and R 3 Selected from or

[0012] R 2 Selected from R 3 Selected from

[0013] R 1 Selected from CH2X, wherein

[0014] 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.

[0015] In some embodiments, the AZD8055 compound is any one of the following chemical structures:

[0016]

[0017]

[0018] Also provided is the use of the above-mentioned AZD8055 compound in preparing a medicament for improving the vigor of crop seeds.

[0019] Also provided is a method for improving crop seed vigor, using the above-mentioned AZD8055 compound to target the seed ABA signaling pathway to improve crop seed vigor.

[0020] Also provided is a method for synthesizing the compound represented by the above general formula (I), which is characterized by comprising the following steps:

[0021] 2,6-dichloro-6-carboxypyridine synthesis

[0022] Will synthesis

[0023] Will Synthesis of the chemical structure of general formula (I)

[0024]

[0025] Beneficial effects of using the AZD8055 compound of the present invention in improving the vigor of crop seeds:

[0026] The present invention discovered that the pyrido[2,3-D]pyrimidine and 4-methoxy-phenyl in the AZD8055 compound can regulate the ABA signaling pathway, thereby effectively enhancing the vigor of various crop seeds, promoting rapid seed germination and robust and consistent emergence. This solves the problem of decreased vigor caused by seed storage and facilitates the successful implementation of direct seeding production. In addition, enhanced seed vigor can greatly double crop yields per mu, bringing huge social benefits and making it suitable for large-scale production and application. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 . This is the effect identification of different concentration treatments of Experimental Example 1 on the germination rate of rice japonica variety Zhonghua 11 seeds.

[0028] Figure 2 This is an evaluation of the effect of the treatment in Experimental Example 1 on the seed vigor of different japonica rice varieties.

[0029] Figure 3 This is an evaluation of the effect of the treatment in Experimental Example 1 on the seed vigor of different indica rice varieties.

[0030] Figure 4 This is the identification of the germination rate of different crop seeds after AZD8055 treatment in Experimental Example 2.

[0031] Figure 5 This is the germination rate test of crop seeds after treatment at different storage periods in Experimental Example 3.

[0032] Figure 6 This is the basic skeleton of the AZD8055 compound shown in Example 1. DETAILED DESCRIPTION

[0033] The preferred embodiments of the present invention will be described in more detail below with reference to the accompanying drawings. Although preferred embodiments of the present invention are shown in the accompanying drawings, it should be understood that the present invention can be implemented in various forms and should not be limited by the embodiments described herein. Rather, these embodiments are provided to make the present invention more thorough and complete and to fully convey the scope of the present invention to those skilled in the art.

[0034] Example 1

[0035] To illustrate the method for obtaining the compound AZD8055 of the present invention, the following contents are disclosed:

[0036] like Figure 6The basic skeleton of the AZD8055 compound shown in the figure, wherein basic skeleton 1 (pyrido[2,3-D]pyrimidine) and basic skeleton 2 (4-methoxy-phenyl) are the basic structures for achieving mTOR inhibition activity and thus regulating seed activity. On this basic skeleton, the 3-position derivatization of the benzene ring of basic skeleton 2 and the 2- and 4-position derivatizations of basic skeleton 1 can better achieve the effect of AZD8055 compound in enhancing seed activity.

[0037] It was found that introducing a hydrophilic group into the R1 group could improve IC 50 To reduce the difficulty of synthesis, R2 and R3 can be designed to have the same structure, or they can be designed to have different structural fragments.

[0038] From this, the following derivative groups were obtained:

[0039]

[0040] Note: The structure within the dotted box can be given priority

[0041] The above symbol "=" means selection.

[0042] To illustrate how to synthesize the basic skeleton of the AZD8055 compound, the following synthesis strategy is described:

[0043] Based on the retrosynthetic method, the analysis is as follows:

[0044]

[0045] Therefore, the synthesis method of the basic skeleton of the AZD8055 compound represented by general formula (I) comprises the following steps:

[0046] 2,6-dichloro-6-carboxypyridine synthesis Will synthesis

[0047] Will Synthesis of the chemical structure of general formula (I)

[0048] Those skilled in the art can implement specific synthesis steps through synthetic experimental means.

[0049] Specifically, the following numbered AZD8055 compounds were obtained, which are only used for labeling purposes:

[0050]

[0051]

[0052] Effect verification:

[0053] To illustrate the effect of the AZD8055 compound of the present invention in enhancing seed vigor, the following experiment was conducted, wherein the AZD8055 compound described in the experimental example is exemplified as follows:

[0054] Drug name: AZD-8055

[0055] CAS No.:1009298-09-2

[0056] Molecular formula: C 25 H 31 N5O4

[0057] Molecular weight: 465.54

[0058] Chemical structure:

[0059]

[0060] AZD8055 compound was used to enhance the vitality of different crop seeds. Details are as follows:

[0061] 1.1 Seed germination experiment: Refer to Wang et al. (2010) with slight modifications. Seeds of promoted varieties, including rice, sweet corn, soybean, tomato, cucumber, water spinach, and Chinese cabbage, as well as their related stockpiled varieties, were tested. Three replicates were set for each material. Fifty mature, plump seeds were selected from each replicate and placed in a 9-cm-diameter, transparent, disposable Petri dish lined with filter paper. 10 mL of distilled water was added, and the Petri dish was then placed in a 28°C constant-temperature, light-incubator (12 h light / 12 h dark) for 5 or 7 days. Germination was defined as when the embryo broke through the seed coat by 2 mm. Seedling establishment was defined as when the radicle reached the seed length and the embryo reached at least half the seed length. The number of germinated and established seeds was counted every 12 h. The germination rate for 5 consecutive days was counted as the 5-day germination potential (GP), and the germination index (GI) = ∑(Gt / Dt) (Dt represents the number of germination days), Gt represents the number of germinated seeds per day corresponding to Dt, and T50 is the time required for the germination rate to reach half (d: day).

[0062] 1.2 Germination experiment with exogenous reagents: For different crops, select the corresponding concentration of reagents from 0μM, 0.2μM, 2μM, 5μM, and 10μM for exogenous application. The germination rate after 3-5 days of application at different concentrations and the germination potential after 5 consecutive days were calculated.

[0063] 1.3 Seedling growth identification experiment:

[0064] (1) Select 50 mature and plump seeds and gently peel off the husks to avoid damaging the embryo.

[0065] (2) Place the shelled seeds in a 50°C oven for about 3 days to break dormancy.

[0066] (3) Then pour the seeds into a 50 mL sterile centrifuge tube, add 10 mL of 75% anhydrous ethanol, and wash the seeds on a shaker at 200 rpm for 5 minutes. Pour off the alcohol in a clean bench (be careful not to drop the seeds), and wash once with sterile water.

[0067] (4) Add 5 mL of 2.5% sodium hypochlorite in a clean hood and wash the seeds in a shaker at 200 rpm for 15-20 minutes. Then, discard the sodium hypochlorite in the clean hood and wash with sterile water 3-5 times.

[0068] (5) Place the washed seeds on sterile filter paper in a clean bench to absorb moisture. Use tweezers that have been sterilized by burning to carefully pick up the dried seeds and place them on a 1 / 2 MS culture medium plate. After inoculation, seal the culture dish with sealing film and place it in a dark incubator at 28°C for 2 days.

[0069] (6) After most of the seeds turn white, they are disinfected with alcohol on the surface and opened in a clean bench. The seeds with the same germination are inoculated on 1 / 2 MS culture medium plates containing reagents of different concentrations. After sealing, they are placed in a constant temperature and light incubator at 28°C (light 12h / dark 12h) and cultured for 12 days.

[0070] (7) Count the length of rice seedlings, root length and number of roots, and record and take photos.

[0071] 1.4 Seedling soil culture growth experiment:

[0072] Three replicates were set up for each material. Thirty mature, plump seeds were selected from each replicate and placed in a 9-cm-diameter, transparent, disposable Petri dish lined with filter paper. 10 mL of distilled water or reagent-treated solution was added, and the dish was placed in a 28°C constant-temperature, light-incubator (12 h light / 12 h dark) for three days. The seeds were then transferred to pots filled with moistened field soil and placed in a 28°C constant-temperature, light-incubator (12 h light / 12 h dark) for 14 days. During this period, the seedlings were observed and photographed.

[0073] Experimental Example 1

[0074] AZD8055 compound treatment concentration screening and testing of different rice varieties:

[0075] Taking the japonica rice variety Zhonghua 11 as an example, AZD8055 was tested to improve seed vigor, and appropriate concentration screening and identification were carried out (Treatment means treatment). The AZD8055 compound was allowed to act on the seeds of the japonica rice variety Zhonghua 11, and then the seeds were placed on a culture dish and waited for germination. Figure 1The results showed that AZD8055 does have the effect of improving seed vigor (germination rate), and the best effect is achieved at a concentration of 2 μM.

[0076] Based on the above results, the same treatment was performed on other different japonica rice varieties (3: EB Gopher; 10: Ao Chiu2Hao; 12: Bombilla; 14: BERLIN) using a 2 μM concentration. Figure 2 The results showed that the germination rate and seedling growth (seedling length) of the treated materials were significantly improved.

[0077] In order to further determine the effect of AZD8055 on improving seed vigor of different rice varieties, different indica rice varieties (6126: Shengyou 6126; 51: Nayou 51; 1179: Ruanhuayou 1179; 6388: Nayou 6388) were treated with the same method at a concentration of 2 μM. Figure 3 The results showed that both could significantly improve the germination rate.

[0078] It can be seen that the AZD8055 compound of the present invention can achieve a significant effect of enhancing seed vigor.

[0079] Experimental Example 2

[0080] Test of AZD8055 compound treatment on different crop seeds

[0081] To further explore the application scope of the AZD8055 compound in seed vigor regulation, relevant tests were conducted on different crop varieties (wheat Ke Nong 199, corn, soybean Dong Nong 47, tomato AC, Chinese cabbage variety Xu Ke Feng Kang 80, red cabbage variety improved Lion, and water spinach variety Xu Ke Jian Ye Kong Kong Cai) using the same treatment steps as in Experimental Example 1. Figure 4 The results showed that the germination rate of treated seeds was significantly higher than that of untreated seeds.

[0082] Experimental Example 3

[0083] Test of AZD8055 compound treatment on crop seeds at different storage periods

[0084] To further explore the application scope of AZD8055 on seed vigor, crop varieties at different storage periods were treated and tested, including the soybean variety Yangchun and the sweet corn varieties Huameitian 16 and Huameitian 8. Figure 5 The results showed that AZD8055 treatment significantly improved seed germination rates in soybean varieties stored for 3, 7, and 10 years; the treatment also promoted germination in sweet corn varieties stored for 2 and 4 years.

[0085] It can be seen that the AZD8055 compound of the present invention can enhance seed vigor. Since the other numbered AZD8055 compounds contain the same skeleton as the numbered 1AZD8055 compound, on the basis of the numbered 1AZD8055 compound's ability to enhance seed vigor, the other numbered AZD8055 compounds, whose derived groups are mainly N, O-containing alkyl or cycloalkyl groups, also have the same effect of enhancing seed vigor.

[0086] The foregoing description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Those skilled in the art will readily appreciate that various modifications and variations of the present invention are possible. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention are intended to be within the scope of protection of the present invention.

Claims

1. The use of the AZD8055 compound in improving the germination rate, germination rate and seedling length of rice seeds. The chemical structural formula of the AZD8055 compound is: ； The concentration of the AZD8055 compound was 2 μM.

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