Use of anticancer drugs for the preparation of a medicament for accelerating winter wheat breeding

Abstract

The application discloses application of an anticancer drug to preparation of an accelerated winter wheat breeding drug and relates to the technical field of winter wheat breeding.The application is characterized in that a histone modification enzyme inhibitor or an O-GlcNAcase (OGA) inhibitor is used to treat winter wheat by seed soaking and / or spraying during planting, so that a vernalization regulation flowering pathway can be specifically activated, and expression of winter wheat vernalization genes such as VRN1 is promoted, thereby replacing the low-temperature vernalization treatment process of winter wheat, significantly reducing the rapid breeding cost of winter wheat, simplifying the rapid breeding cultivation mode, and improving the rapid breeding efficiency.The application uses the anticancer drug to treat winter wheat, so that the low-temperature vernalization treatment is not needed in the winter wheat breeding process, and the winter wheat can be planted under a conventional greenhouse (RG) condition (20 DEG C; 16h light: 8h darkness) to realize early heading and flowering, and the breeding period of the winter wheat is significantly shortened.

Description

Technical Field

[0001] This invention relates to the field of winter wheat breeding technology, specifically to the application of anticancer drugs as preparation of drugs to accelerate winter wheat breeding. Background Technology

[0002] Traditional wheat breeding processes require low-temperature vernalization treatment of wheat seeds. Based on the different vernalization temperatures and durations at different stages of vernalization, wheat varieties can be classified into winter wheat and spring wheat.

[0003] Generally, the vernalization temperature for spring wheat is 0-12℃, and the vernalization period is 5-15 days. Spring wheat is not sensitive to temperature, and seeds sown without vernalization treatment can still head and set normally in spring. The vernalization temperature for winter wheat is 0-7℃, and the vernalization period is between 15-70 days. Winter wheat that has not undergone vernalization treatment will have delayed heading, or may even fail to head.

[0004] Jin-Kyung Cha et al., in their paper "Speed ​​vernalization to accelerate generation advance in winter cereal crops," demonstrated that adjusting the photoperiod of winter wheat can effectively shorten vernalization time, thereby shortening the heading and grain-filling periods and accelerating winter wheat breeding. However, this study still requires germinating winter wheat at 4°C first, followed by long-day vernalization at a lower temperature (8°C-10°C) for 4 weeks. Therefore, current research still requires certain low-temperature vernalization treatments for winter wheat to accelerate winter wheat breeding. Summary of the Invention

[0005] Based on this, the present invention provides a method for accelerating winter wheat breeding. This method involves soaking winter wheat seeds with existing anti-tumor drugs and / or spraying them during the planting process, thereby enabling winter wheat to achieve rapid heading and grain filling without the need for low-temperature vernalization treatment, thus accelerating winter wheat breeding.

[0006] In some embodiments, this invention utilizes chemical OGA inhibitors (O-GlcNAc hydrolase inhibitors) to treat winter wheat seeds through soaking and / or spraying during the plant growth stage, thereby altering the O-GlcNAc signaling in winter wheat and increasing the plant's monosaccharide levels. This allows winter wheat to accelerate heading and grain filling without the need for vernalization, achieving the goal of rapid winter wheat breeding. This invention utilizes OGA inhibitors to treat winter wheat, enabling it to achieve heading and grain filling without vernalization, providing a new solution for accelerating winter wheat breeding.

[0007] In some embodiments, the present invention utilizes histone modification inhibitors (histone deacetylase inhibitors) to soak winter wheat seeds and / or spray them during the growth stage of winter wheat plants to accelerate the heading and grain filling of winter wheat, without the need for low-temperature vernalization of winter wheat, thereby achieving the purpose of rapid breeding.

[0008] The OGA inhibitor used in this invention is at least one of tiamet G and PUGNAc.

[0009] The histone modification inhibitor used in this invention is at least one of Trichostatin A (TSA), Panobinostat, Tucidinostat, and Belinostat.

[0010] Compared with the prior art, the present invention has the following beneficial effects:

[0011] This invention utilizes diluted anticancer drugs to treat wheat, thereby specifically activating the expression of vernalization regulatory genes such as VRN1 in winter wheat. This accelerates the heading, flowering, and grain-setting processes of winter wheat, shortening the wheat breeding cycle. By treating winter wheat with anticancer drugs, low-temperature vernalization treatment is no longer required during winter wheat breeding. Winter wheat can be rapidly bred under conventional greenhouse (RG) conditions (20℃; 16h light: 8h darkness), significantly shortening the winter wheat breeding cycle and simplifying the planting method. Attached Figure Description

[0012] Figure 1 The growth of winter wheat after 80 days of planting is shown in blank example 1 (left) and example 1 (right).

[0013] Figure 2 The growth of winter wheat after 85 days of planting is shown in blank example 2 (left) and example 6 (right);

[0014] Figure 3 Example 1: The relative expression level of the VRN1 gene before and after soaking in PUGNAc seeds of Kenong 199 winter wheat.

[0015] Figure 4 Example 9: The relative expression levels of the VRN1 gene before and after soaking in Thiamet G for the winter wheat variety Kenong 199.

[0016] Specific implementation methods

[0017] The present invention will be further described below with reference to specific embodiments. The winter wheat seeds and anticancer drugs used in the following embodiments of the present invention are all commercially available products.

[0018] The experimental materials used in this invention are as follows:

[0019] Winter wheat varieties include Jing 411, Jingdong 8, Bainong Aikang 58, and Kenong 199 (KN199).

[0020] PUGNAc (Molecular formula: C 15 H 19 N3O7), purity ≥95%, Merck Chemicals

[0021] Thiamet G (molecular formula: C9H) 16 N2O4S), purity ≥98%, Merck Chemicals

[0022] Dimethyl sulfoxide (DMSO), analytical grade

[0023] TSA (Trichostatin A, C) 17 H 22 N2O3), purity ≥98%, Merck Chemicals

[0024] Panobinostat (molecular formula: C) 21 H 23 N3O2), purity ≥98%, Merck Chemicals

[0025] Tucidinostat (molecular formula: C) 22 H 19 FN4O2), purity ≥98%, Doppler

[0026] Belinostat (molecular formula: C) 15 H 14 N2O4S), purity ≥98%, Merck Chemicals

[0027] GSK126 (methyltransferase inhibitor, molecular formula: C) 31 H 38 N6O2), purity ≥98%, Beijing Bailingwei Technology Co., Ltd.

[0028] GSK-J1 (demethylase inhibitor, molecular formula: C) 22 H 23 N5O2), purity ≥99%, Abdominal Pharma Technology (Shanghai) Co., Ltd.

[0029] Torin2 (mTOR inhibitor, molecular formula: C) 24 H 15 F3N4O), purity ≥98%, Merck Chemicals

[0030] Iadademstat (an LSD1 inhibitor, molecular formula: C) 15 H22 N2)

[0031] Example 1

[0032] A method for accelerating winter wheat breeding, comprising the following steps:

[0033] 5 mg of PUGNAc was dissolved in 3 mL of dimethyl sulfoxide to obtain a stock solution, which was then diluted with water to prepare drug solutions of different concentrations (20 μM-20 nM).

[0034] The above-mentioned concentration of drug solution was used as the soaking solution to soak the winter wheat of Kenong 199 at room temperature for 3 days. After the seeds showed signs of sprouting, they were planted in the soil and then cultivated in a conventional greenhouse under conventional greenhouse (RG) conditions (20℃; 16h light: 8h darkness).

[0035] One week after winter wheat sowing (when the wheat plants are 5-8 cm tall), spray the roots of the winter wheat with the above-mentioned high-concentration pesticide solution at a rate of 1 mL / cm². 2 Spray once a week for four consecutive weeks; then spray the roots of winter wheat with a low-concentration pesticide solution at a rate of 1 mL / cm². 2 Spray twice a week until the plants begin to elongate, then stop spraying. Ensure sufficient water and fertilizer during greenhouse cultivation and record the heading time.

[0036] Blank Example 1: The winter wheat seed of Kenong 199 was soaked in water, and the above-mentioned drug solution of the present invention was not sprayed during the planting process.

[0037] Examples 2-5

[0038] The concentration of the soaking solution in Example 1 was adjusted while other parameters remained unchanged. The effect of the concentration of PUGNAc in the soaking solution on the heading time of Kenong 199 winter wheat was investigated, as shown in Table 1.

[0039] Table 1

[0040]

[0041]

[0042] Examples 6-8

[0043] Using the method of Example 1, the only difference was that the winter wheat variety Kenong 199 was replaced with Jingdong No. 8, Bainong Aikang 58, and Jing 411 respectively, while all other varieties remained the same. The heading time was recorded as shown in Table 2.

[0044] Table 2

[0045] Winter wheat variety Planting time / day when beginning to head Planting time / day when completely heading Example 1 Kanon 199 84.25±0.5 89.5±0.58 Example 6 Jingdong 8 113.75±0.96 118.5±0.57 Blank Example 2 Jingdong 8 120.5±0.5 124.75±1.26 Example 7 Bainong Dake 58 106.75±1.5 113±1.41 Blank Example 3 Bainong Dake 58 113.5±1.29 118.25±0.96 Example 8 Jing 411 106.75±0.5 112.75±1.5 Blank Example 4 Jing 411 111.25±0.96 116.75±0.96

[0046] Note: Blank Example 2 in Table 2 represents Jingdong No. 8 winter wheat that was not treated with PUGNAc soaking and spraying;

[0047] Blank Example 3 is Bainong Dwarf Resistant 58 winter wheat that was not treated with PUGNAc soaking or spraying;

[0048] Blank Example 4 shows winter wheat of variety 411 that was not treated with PUGNAc for soaking and spraying.

[0049] Examples 9-16

[0050] A method for accelerating winter wheat breeding, referring to Example 1, differs only in that Thiamet G, TSA, Panobinostat, Tucidinostat, Belinostat, GSK126, GSK-J1, Torin2, and Iadademstat are used to replace PUGNAc in Example 1, that is, other drugs are used to treat the Kenong 199 winter wheat (seed soaking and spraying). The specific treatment method is the same as in Example 1, and the heading time is recorded as shown in Table 3.

[0051] Table 3

[0052]

[0053]

[0054] from Figure 1 and Figure 2 As can be seen from the above, after winter wheat is treated with PUGNAc in Example 1 of the present invention, it can cause winter wheat to head earlier and shorten the breeding cycle.

[0055] from Figure 3 Example 1: The relative expression levels of the VRN1 gene in Kenong 199 winter wheat before and after soaking with PUGNAc showed that the VRN1 expression level of Kenong 199 winter wheat was significantly increased after soaking with PUGNAc.

[0056] from Figure 4 Example 9: The relative expression levels of the VRN1 gene in Kenong 199 winter wheat before and after soaking with Thiamet G showed that the VRN1 expression level of Kenong 199 winter wheat was significantly increased after soaking with Thiamet G.

[0057] It should be noted that the above examples are merely some specific embodiments of the present invention. Obviously, the present invention is not limited to the above embodiments and other modifications are possible. All modifications directly or indirectly derived by those skilled in the art from the disclosure of the present invention should be considered within the scope of protection of the present invention.

Claims

1. The application of anticancer drugs in the preparation of drugs to accelerate winter wheat breeding, characterized in that, The anticancer drug is at least one of histone modification inhibitors or OGA inhibitors; The OGA inhibitor is at least one of tiotropium bromide G and PUGNAc; The histone modification inhibitor is at least one of palbistat, chidamide, and belistat.

2. The application according to claim 1, characterized in that, The anticancer drug can be diluted and used for soaking winter wheat seeds, and / or the anticancer drug can be diluted and used as a spray on winter wheat plants.

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