Application of AZD8055 compound in improvement of crop seed vigor

Abstract

The invention relates to the technical field of regulation and control of crop seed vigor, in particular to application of an AZD8055 compound in improvement of crop seed vigor, the AZD8055 compound comprises the following chemical structure: # imgabs0 #, and the AZD8055 compound can effectively improve the seed vigor and promote rapid germination of seeds, so that seedlings emerge robustly and consistently.

Description

Technical Field

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

[0002] Seed vitality is the core indicator for measuring seed quality and efficacy, and has a profound impact on the high-quality development of modern agricultural production. The level of seed vitality not only directly affects the yield and quality of crops, but also determines the emergence effect after sowing, the robustness of seedling growth, and subsequent production potential. High-quality, high-vitality seeds show excellent storage resistance during storage, and can germinate quickly and evenly under suitable conditions to form a uniform emergence pattern, which is crucial to the successful implementation of direct seeding methods for crops such as rice and corn. It also plays a decisive role in optimizing field planting density, enhancing the competitive advantage between crops and weeds, and improving the ability to resist the invasion of pests and diseases and stress. International data shows that about one-fifth of the global seed market suffers huge economic losses due to challenges such as uneven emergence, weak adversity response, and poor seedling growth caused by seed vitality problems.

[0003] In view of this, the research and development and application of seed vitality enhancement technology has become the key to achieving goals such as increasing yields and reducing inventory costs. The transformation process of seeds from dormancy to germination is the core step that reflects its vitality properties. It is also a landmark event at the beginning of the plant life cycle and has an irreplaceable standard status for judging the quality of seeds.

[0004] Studies have found that the process of seed dormancy and germination is a complex life phenomenon, involving gene expression regulation, environmental signal sensing, and the synergistic effects of multiple biochemical pathways. Among them, the plant hormone abscisic acid (ABA) plays a pivotal role in the regulation of seed vigor. As an important hormone widely present in vascular plants, ABA is involved in the entire life cycle of seeds from development to maturity, including but not limited to physiological processes such as dormancy induction, germination initiation, stomatal movement regulation, fruit ripening control, and various adversity responses. Taking rice as an example, 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, while the changes in the expression levels of the two are directly affected by the concentration of ABA, indicating that the ABA signaling pathway may finely adjust seed vigor through the bZIP23-PER1A signaling axis. During the seed maturity stage, the endogenous concentration of ABA rises to a peak value, and by finely regulating the accumulation of reserve substances such as fat, starch, and storage protein, it prevents seeds from germinating prematurely and maintains their dormant state. The latest study also revealed the mechanism by which OsUGT75A accelerates seed germination by reducing ABA content. Accordingly, stored seeds generally accumulate higher ABA concentrations, which inhibit seed germination through its signal 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, it was revealed that the ubiquitination system is involved in the degradation process of ABA pathway regulatory factors ABA Insensitive 3 (ABI3) and ABI5 proteins, but currently more focus is on the proteasome degradation mechanism mediated by the ubiquitination pathway, and 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 provide a use of an AZD8055 compound in improving the vigor of crop seeds, wherein the AZD8055 compound can effectively enhance seed vigor, promote rapid seed germination, and subsequently produce robust and uniform seedlings.

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

[0008] Provided is the use of the AZD8055 compound in improving the vigor of crop seeds. 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 CH 2 X, where

[0014] X is selected from OH, NHCH 2 CH 2 OH, NHCH 2 CH 2 OMe, N(CH 2 CH 2 OH) 2 , NHCH 2 CH 2 NHMe, NHCH 2 CH 2 NMe 2 , NHCO(OR), NHCH 2 CO 2 Et, NHCH 2 CO 2 H, NHMe, NMe 2 , or R 1 Selected from CONH 2 ,CONHOH,CONHMe,CONHCH 2 CH 2 OH,CONHCH 2 CH 2 OMe, CONHCH 2 CH 2 NHMe, CSNH 2 .

[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 AZD8055 compound in preparing a medicine for improving the vitality of crop seeds.

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

[0020] Also provided is a method for synthesizing the compound represented by the above general formula (I), characterized in that it comprises 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 the use of the AZD8055 compound of the present invention in improving the vitality of crop seeds:

[0026] The present invention discovered that pyrido [2,3-D] pyrimidine and 4-methoxy-phenyl in the AZD8055 compound can achieve regulation of the ABA signaling pathway, thereby effectively enhancing the vigor of various crop seeds, promoting rapid germination of seeds and robust and uniform emergence of seedlings, thereby solving the problem of decreased vigor caused by seed storage and facilitating the successful implementation of direct seeding production. Furthermore, enhanced seed vigor can greatly promote a doubling of crop yield per mu, bringing huge social benefits, and is suitable for large-scale production and application. BRIEF DESCRIPTION OF THE DRAWINGS

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

[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 germination rate identification 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. Figure 6 This is the basic skeleton of the AZD8055 compound shown in Example 1. DETAILED DESCRIPTION

[0032] The preferred embodiments of the present invention will be described in more detail below with reference to the accompanying drawings. Although the 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. On the contrary, 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.

[0033] Example 1

[0034] To illustrate the method for obtaining the compound AZD8055 of the present invention, the following contents are disclosed:

[0035] like Figure 6 The basic skeleton of the AZD8055 compound shown in the figure, wherein the basic skeleton 1 (pyrido [2,3-D] pyrimidine) and the basic skeleton 2 (4-methoxy-phenyl) are the basic structures for achieving mTOR inhibition activity and thus regulating seed activity, and on this basic skeleton, the 3-position derivatization of the benzene ring of the basic skeleton 2 and the 2- and 4-position derivatization of the basic skeleton 1 can better achieve the effect of the AZD8055 compound in enhancing seed activity.

[0036] It was found that introducing a hydrophilic group into the R1 group could improve IC 50 To reduce the difficulty in synthesis, R2 and R3 can be designed to have the same structure, and of course R2 and R3 can be designed to have different structural fragments.

[0037] From this, the following derivative groups were obtained:

[0038] Note: The structure in the dotted box can be given priority

[0039] The above symbol "=" means selection.

[0040] In order to illustrate how to synthesize the basic skeleton of AZD8055 compound, the synthesis idea is explained as follows:

[0041] Based on the retrosynthetic method, the analysis is as follows:

[0042]

[0043] Therefore, the synthesis method of the basic skeleton of the AZD8055 compound shown in general formula (I) comprises the following steps:

[0044] 2,6-Dichloro-6-carboxypyridine synthesis

[0045] Will synthesis

[0046] Will Synthesis of the chemical structure of general formula (I)

[0047] Those skilled in the art can implement specific synthesis steps through synthetic experimental means.

[0048] Specifically, the following numbered AZD8055 compounds were obtained, and the numbers are only used for labeling purposes:

[0049]

[0050]

[0051] Effect verification:

[0052] In order to illustrate the effect of the AZD8055 compound of the present invention in improving seed vigor, the following experiment was conducted, wherein the AZD8055 compound described in the experimental example is exemplified as:

[0053] Drug name: AZD-8055

[0054] CAS No.:1009298-09-2

[0055] Molecular formula: C 25 H 31 N 5 O 4

[0056] Molecular weight: 465.54

[0057] Chemical Structure:

[0058]

[0059] AZD8055 compound was used to improve the vitality of seeds of different crops. Details are as follows:

[0060] 1.1 Seed germination experiment: refer to Wang et al. (2010) with slight modifications. Seeds of promoted varieties such as rice, sweet corn, soybean, tomato, cucumber, water spinach, Chinese cabbage and their related stockpiles were used as test objects. Three replicates were set for each material. 50 mature and plump seeds were selected for each replicate and placed in a transparent disposable culture dish with a diameter of 9 cm covered with filter paper. 10 mL of distilled water was added, and then the culture dish was placed in a 28°C constant temperature and light incubator (light 12 h / dark 12 h) for 5 or 7 days. The germination standard was when the embryo broke through the seed coat by 2 mm, and the seedling standard was when the radicle reached the length of the seed and the embryo reached more than half of the length of the seed. The number of seeds that germinated and seedled was counted every 12 hours. The germination rate for 5 consecutive days was counted as the 5-day germination potential (GP), 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).

[0061] 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 treatment. The germination rate of 3-5 days under different concentration conditions and the germination potential for 5 consecutive days are statistically analyzed.

[0062] 1.3 Seedling growth identification experiment:

[0063] (1) Select 50 mature and plump seeds and gently peel off the husks, trying to avoid damaging the embryo.

[0064] (2) Place the shelled seeds in a 50°C oven for about 3 days to break dormancy.

[0065] (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 min. Pour off the alcohol in a clean bench (be careful not to drop the seeds), and wash once with sterile water.

[0066] (4) Add 5 mL of 2.5% sodium hypochlorite in a clean bench and wash the seeds on a shaker at 200 rpm for 15-20 min. Then pour out the sodium hypochlorite in the clean bench and wash with sterile water for 3-5 times.

[0067] (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 / 2MS culture medium plate. After inoculation, seal the culture dish with sealing film and place it in a 28°C incubator in the dark for 2 days.

[0068] (6) After most of the seeds have turned white, they are disinfected with alcohol on the surface and opened in a clean bench. Seeds with consistent germination are inoculated onto 1 / 2MS culture medium plates containing reagents of different concentrations. After sealing, they are placed in a 28°C constant temperature and light incubator (12 h light / 12 h dark) for 12 days.

[0069] (7) Count the length of rice seedlings, root length and number of roots, and record and take photos.

[0070] 1.4 Seedling soil culture growth experiment:

[0071] Three replicates were set for each material. Thirty mature and plump seeds were selected for each replicate and placed in a 9-cm diameter transparent disposable culture dish covered with filter paper. 10 mL of distilled water or reagent treatment solution was added, and then the culture dish was placed in a 28°C constant temperature and light incubator (12 h of light / 12 h of darkness) for 3 days. The seeds were then transferred to a pot filled with moist field soil and placed in a 28°C constant temperature and light incubator (12 h of light / 12 h of darkness) for 14 days. During this period, the growth of the seedlings was observed and recorded and photographed.

[0072] Experimental Example 1

[0073] AZD8055 compound treatment concentration screening and testing of different rice varieties:

[0074] Taking the japonica rice variety Zhonghua 11 as an example, AZD8055 was tested for improving seed vitality, 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 1 The results showed that AZD8055 did have the effect of improving seed vigor (germination rate), and the best effect was achieved at a concentration of 2 μM.

[0075] 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 concentration of 2 μM. Figure 2 The results showed that the germination rate and seedling growth (seedling length) of the treated materials were significantly improved.

[0076] 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 concentration of 2 μM. Figure 3 The results showed that both could significantly improve the germination rate.

[0077] It can be seen that the AZD8055 compound of the present invention can achieve a significant effect of enhancing seed vigor.

[0078] Experimental Example 2

[0079] Test of AZD8055 compound treatment on seeds of different crops

[0080] In order to further develop the application scope of AZD8055 compound in seed vigor regulation, relevant tests were carried out 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 King, and water spinach variety Xu Ke Jian Ye Kong Xin Cai) with the same processing 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.

[0081] Experimental Example 3

[0082] Testing of AZD8055 compound treatment on crop seeds at different storage periods

[0083] In order to further explore the application scope of AZD8055 on seed vigor, crop varieties at different storage periods were treated and tested, including soybean variety Yangchun and sweet corn varieties Huameitian 16 and Huameitian 8. Figure 5 The results showed that AZD8055 treatment could significantly improve the seed germination rate of soybean varieties stored for 3, 7 and 10 years; the treatment also showed the effect of promoting germination in sweet corn varieties stored for 2 and 4 years.

[0084] It can be seen that the AZD8055 compound of the present invention can improve 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 having the ability to improve seed vigor, the derived groups of the other numbered AZD8055 compounds are mainly alkyl or cycloalkyl containing N, O, and also have the same effect of improving seed vigor.

[0085] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. Use of the AZD8055 compound in improving the vigor of crop seeds, wherein the AZD8055 compound comprises a chemical structure represented by general formula (I): in, R 2 and R 3 Selected from or R 2 Selected from R 3 Selected from R 1 Selected from CH2X, wherein 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.

2. The use of the AZD8055 compound according to claim 1 in improving the vigor of crop seeds, characterized in that: The AZD8055 compound is any one of the following chemical structures:

3. Use of the AZD8055 compound according to claim 1 or 2 in the preparation of a medicament for improving the vigor of crop seeds.

4. A method for improving the vitality of crop seeds, characterized in that: The AZD8055 compound described in claim 1 or 2 is used to target the seed ABA signaling pathway to improve crop seed vigor.

5. A method for synthesizing the compound represented by the general formula (I) according to claim 1 or 2, characterized in that: The following steps are involved: 2,6-Dichloro-6-carboxypyridine synthesis Will synthesis Will Synthesis of the chemical structure of general formula (I)

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