Application of a sprouting lactone to promote growth of rice under saline-alkali stress conditions

Abstract

The application of sprout lactone in promoting the growth of rice under saline-alkali stress conditions, the application of sprout lactone is disclosed.The application of sprout lactone in promoting the growth of rice under saline-alkali stress conditions, 0.1-10 μmol / L sprout lactone is used for promoting the growth of rice under saline-alkali stress conditions.The sprout lactone can be used as a plant growth regulator, can improve the salt tolerance and alkali stress resistance of rice, and can promote the growth of rice, that is, the germination and growth of the later seedling under stress regulation.The application of sprout lactone in promoting the growth of rice under saline-alkali stress conditions, through the application of low-concentration sprout lactone, the positive effect of sprout lactone on the growth and development of rice is illustrated, which breaks the existing understanding of sprout lactone to a certain extent-the inhibition effect on animal tumor cells, and provides a new application of sprout lactone.

Description

TECHNICAL FIELD

[0001] The present application relates to the application of germinone. BACKGROUND

[0002] Germinone (English name Germinone A, for short Ger), chemical name 3-Methyl-5-((3-nitro-[1,1'-biphenyl]-4-yl)oxy)furan-2(5H)-one (CAS T9007) chemical structure formula. It is an artificially synthesized chemical substance, and whether the substance exists in natural products has not been reported.

[0003] The existing function research of germinone mainly focuses on animal research. The results of animal and cell in vitro experiments show that germinone (code CRT0066101) shows inhibitory activity on tumor cells in a variety of tumor cells, and the target protein is protein kinase D (Protein kinase D). By inhibiting the G2 / M process of the cell cycle, the inhibitory effect on tumor cells is realized.

[0004] The soil of saline-alkali land contains more salt and alkaline components, and the growth of rice suffers from various adverse effects of salt-alkali stress. The damage of salt-alkali stress to plants includes osmotic stress, ion toxicity and alkali stress (high pH stress), which is more serious than any single stress. At present, the measures to improve the stress resistance and yield of rice in saline-alkali land include improving the soil, screening and breeding salt-tolerant varieties and molecular breeding methods, but there are problems such as high improvement cost, long cycle and so on, and there is still much potential for technical improvement and upgrading. Among them, the addition of exogenous substances can quickly induce the self-induced effect of rice to improve the salt-tolerance of rice, and the combined application of a variety of substances can have a synergistic effect, which becomes an effective way to improve the yield of rice in soda saline-alkali land. SUMMARY

[0005] The present application provides an application of germinone for promoting the growth of rice under salt-alkali stress conditions, a new application of germinone.

[0006] The application of germinone for promoting the growth of rice under salt-alkali stress conditions provided by the present application, 0.1-10 μmol / L germinone is used for promoting the growth of rice under salt-alkali stress conditions.

[0007] The germinolide can be used as a plant growth regulator, can improve the salt tolerance and alkali stress tolerance of rice, and can promote the growth of rice, i.e., promote the germination of rice seeds and the growth of later seedlings under stress regulation. Low concentration 0.1-10 μmol / L germinolide can alleviate the activity of rice seeds under salt and alkali stress; low concentration 0.1-10 μmol / L germinolide can significantly alleviate the inhibition of the growth of rice seedlings under salt and alkali stress, including the growth and development of the aboveground part and the root. The application of the present application can accelerate the growth of seedlings under salt and alkali stress conditions (such as saline-alkali land), shorten the plant seedling period, reduce the seedling mortality rate, improve the production efficiency, and save the production cost.

[0008] The application of the germinolide of the present application in promoting the growth of rice under salt and alkali stress illustrates the positive effect of the germinolide on the growth and development of rice by low concentration germinolide application, to some extent, breaks the existing understanding of the germinolide by most people-inhibition of animal tumor cells, and provides a new application of the germinolide. The present application can lay a good application foundation for promoting the growth of seedlings in agricultural production, and further provide a direction for genetic engineering to improve plants. BRIEF DESCRIPTION OF DRAWINGS

[0009] Figure 1 The structural formula of the germinolide of the present application is shown in the following formula:

[0010] Figure 2 The germinolide of Example 1 respectively improves the salt and alkali stress tolerance of rice seeds, and the comparison chart is shown in the following figure:

[0011] Figure 3 The germinolide of Example 1 respectively affects the germination rate of rice under salt and alkali stress, and the result chart is shown in the following figure:

[0012] Figure 4 The germinolide of Example 2 respectively alleviates the growth of rice seedlings under salt and alkali stress, and the comparison chart is shown in the following figure:

[0013] Figure 5 The germinolide of Example 2 respectively affects the length of the aboveground part of rice under salt and alkali stress, and the result chart is shown in the following figure. DETAILED DESCRIPTION

[0014] The technical solutions in the embodiments of the present application will be described below in conjunction with the embodiments of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0015] It should be noted that the embodiments in the present application and the features in the embodiments can be combined with each other without conflict.

[0016] Specific embodiment one: the application of germinone in promoting the growth of rice under saline-alkaline stress conditions, 0.1-10 μmol / L germinone is used to promote the growth of rice under saline-alkaline stress conditions.

[0017] The germinone (English name Germinone A, Ger for short) described in this embodiment has a chemical name of 3-Methyl-5-((3-nitro-[1,1'-biphenyl]-4-yl)oxy)furan-2(5H)-one (CAS T9007), and a chemical structural formula as shown in the figure. Figure 1 Animal and cell in vitro experimental results show that germinone (code CRT0066101) shows an inhibitory anti-tumor cell activity in a variety of tumor cells, and the target protein is protein kinase D (Protein kinase D). By inhibiting the G2 / M process of the cell cycle, the inhibitory effect on tumor cells is achieved. Recent studies in plants have found that germinone and desmethyl-type germinone (dMGer) are dependent on the presence of KAI2 protein and inhibit the elongation of the hypocotyl.

[0018] As a plant growth regulator, germinone in this embodiment can bind to its receptor KAI2 to transmit downstream signals, activate the scavenging system of active oxygen, and at the same time protect tissues from oxidative stress; in addition, the KAI2 signal can also interact with the plant hormone IAA, and IAA can promote the growth of plant stems and the elongation of roots; low concentration (0.1-10 μM) germinone (Ger) promotes the germination of rice seeds and the growth of seedlings under saline-alkaline stress conditions. The seed germination rate and emergence speed of rice in saline-alkaline land are improved, thereby accelerating the growth of seedlings, shortening the plant seedling period, reducing the mortality rate of seedlings, improving production efficiency, and saving production costs.

[0019] Specific embodiment two: the difference between this embodiment and specific embodiment one is that the application method of germinone in promoting the growth of rice under saline-alkaline stress conditions is carried out according to the following steps:

[0020] I. Pre-treatment of rice seeds: the rice seeds are fully suspended and soaked in 75% ethanol for 10 min, and then fully suspended and soaked in 0.02% mercuric chloride for 10 min. After taking out the rice seeds, they are repeatedly rinsed with distilled water for 10 times, then distilled water is added to cover the rice seeds, and the mixture is placed in a constant temperature box for 24 h to obtain treated rice seeds.

[0021] II. The treated rice seeds are placed in a 10 cm diameter culture dish with filter paper at the bottom. 2 ml of germinant lactones with a concentration of 0.1-10 μmol / L are added to the culture dish for cultivation. The germinant lactones are replaced every 2 days, and the replacement is performed 1-6 times, thereby achieving the application of the germinant lactones.

[0022] In step one of the embodiment, the temperature of the thermostat is 30°C.

[0023] In step two of the embodiment, the culture method is as follows: the culture is performed in a growth chamber with 12 h light at 28°C and 12 h darkness at 22°C.

[0024] The 1-10 μmol / L (μM) germinant lactones can relieve the toxicity of NaCl and Na2CO3 to rice, and further improve the tolerance of rice seeds and seedlings to salt and alkali stress.

[0025] Example 1: Germinant lactones promote the germination of rice seeds under salt and alkali stress

[0026] The specific steps are as follows:

[0027] I. Pretreatment of rice seeds: rice (Oryza sativa, variety 'Dongdao 122') seeds are fully suspended in 75% ethanol for 10 min, and then fully suspended in 0.02% mercuric chloride for 10 min. The rice seeds are taken out and rinsed with distilled water for 10 times. Distilled water is then added to cover the rice seeds, and the mixture is placed in a 30°C thermostat for 24 h to obtain treated rice seeds.

[0028] 2. Place the rice seeds treated in step one into a 10cm diameter petri dish, with two sheets of filter paper at the bottom. Add 2 ml of an experimental solution with a concentration of 0.1–10 μmol / L to the petri dish for incubation. Change the experimental solution every 2 days, for a total of 3 times. The experimental solution is as follows: MOCK (with an equal amount of dimethyl sulfoxide), 200 mM (mmol / L) NaCl, 50 mM (mmol / L) Na2CO3, 0.1 μM Ger + 200 mM (mmol / L) NaCl, 1 μM Ger + 200 mM (mmol / L) NaCl, 5 μM Ger + 200 mM (mmol / L) NaCl, 10 μM Ger + 200 mM (mmol / L) NaCl, 0.1 μM Ger + 50 mM (mmol / L) Na2CO3, 1 μM Ger + 50 mM (mmol / L) Na2CO3, 5 μM Germ + 50mM (mmol / L) Na2CO3 and 10μM Ger + 50mM (mmol / L) Na2CO3 were used. The petri dishes containing rice seeds and the experimental solution were placed in a growth chamber at 28℃ for 12 hours of light and 22℃ for 12 hours of darkness for cultivation. The germination of the seeds was observed and the germination rate was recorded every day. Photos were taken when necessary.

[0029] In this embodiment, step two sets different germination lactone concentrations, where 0 represents MOCK treatment with an equal amount of dimethyl sulfoxide (DMSO). The different germination lactone concentrations are specifically 0.1 μM (μmol / L), 1 μM (μmol / L), 5 μM (μmol / L), and 10 μM (μmol / L).

[0030] In step two of this embodiment, the experimental solution contains germinating lactone. Since the germinating lactone is produced by the mother liquor organic solvent dimethyl sulfoxide (DMSO), an equal amount of dimethyl sulfoxide should also be added in simple salt and alkali treatments.

[0031] The experimental results are as follows:

[0032] In Example 1, rice seeds were photographed after absorbing water for 5 days. The results are as follows: Figure 2 , Figure 3 As shown. Figure 2 A comparative graph showing the improvement of rice seed tolerance to salt and alkali stress by different concentrations of germinating lactones. Figure 2 The upper part (A) shows the germination of seeds after treatment with 200mM NaCl and exogenous addition of 0.1-10μM germination lactone (Ger); Figure 2 The following section (B) shows the germination of seeds after treatment with 50 mM Na2CO3 and the exogenous addition of 0.1–10 μM germination lactone (Ger); Figure 3Figure 1 shows the effect of different concentrations of germin on the germination rate of rice seeds under salt stress. Figure 3 Figure 1A shows the germination rate of rice seeds under salt stress. MOCK represents the control group, NaCl 200 mM represents the salt treatment group, Ger 0.1 μM + NaCl 200 mM represents the salt treatment group, Ger 1 μM + NaCl 200 mM represents the salt treatment group, Ger 5 μM + NaCl 200 mM represents the salt treatment group, Ger 10 μM + NaCl 200 mM represents the salt treatment group. Figure 3 Figure IB shows the germination rate of rice seeds under alkali stress. MOCK represents the control group, Na2CO3 50 mM represents the alkali treatment group, Ger 0.1 μM + Na2CO3 50 mM represents the alkali treatment group, Ger 1 μM + Na2CO3 50 mM represents the alkali treatment group, Ger 5 μM + Na2CO3 50 mM represents the alkali treatment group, Ger 10 μM + Na2CO3 50 mM represents the alkali treatment group. Figure 3 The data shown in Figure 1 are the average values ± standard deviation of 5 replicates. Significant differences (p < 0.05, two-way ANOVA with LSD multiple comparison test) are indicated by different lowercase letters. From Figure 3 It can be seen from Figure 1A that 200 mM NaCl significantly inhibited the germination of rice seeds, and the exogenous application of 0.1-5 μM germin could alleviate the inhibition of seed germination under 200 mM NaCl stress. From Figure 3 It can be seen from Figure IB that 50 mM Na2CO3 significantly inhibited the germination of rice seeds, and the exogenous application of 0.1-5 μM germin could alleviate the inhibition of seed germination under 50 mM Na2CO3 stress. Although 0.1-10 μM germin could alleviate the salt and alkali stress to some extent, the promotion effect of 1 and 5 μM germin was the most significant, and they were the best concentrations suitable for the germination of rice seeds.

[0033] Example 2: Germin alleviates the growth of rice seedlings under salt and alkali stress

[0034] The specific steps are as follows:

[0035] I. Rice seed pretreatment: Soak rice (Oryza sativa variety 'Dongdao 122') seeds in 75% ethanol for 10 minutes, then soak them in 0.02% mercuric chloride for 10 minutes. After removing the rice seeds, rinse them repeatedly with distilled water 10 times, then fill them with distilled water and place them in a 30℃ constant temperature incubator for 24 hours.

[0036] 2. Place the sterilized rice seeds from step one into disposable plastic petri dishes with a diameter of 10cm. Two layers of filter paper are placed at the bottom of the petri dishes. The experimental solution added is the same as in Example 1. Place the petri dishes containing the rice seeds and experimental solution into a growth chamber at 28℃ for 12 hours of light followed by 12 hours of darkness at 22℃ for 6 days. Then, photograph and measure the length of the seedlings.

[0037] The experimental results are as follows:

[0038] Figure 4 This is a comparison chart showing the effect of germination lactones in alleviating salt and alkali stress on rice seedling growth in this embodiment. Figure 4 In the figure, A represents the germination of seeds after treatment with 200 mM NaCl (salt treatment) and exogenous addition of 0.1–10 μM germination lactone (Ger); Figure 4 B in the figure represents the germination of seeds treated with 50 mM Na2CO3 (alkali treatment) and then exogenously supplemented with 0.1–10 μM germination lactone (Ger). Figure 5 The figure shows the effect of germination lactone on the length of the aboveground parts of rice under salt and alkali stress in this embodiment (statistics on the length of the aboveground parts of seedlings).

[0039] from Figure 4 and Figure 5 It was found that both 200 mM NaCl and 50 mM Na2CO3 could significantly inhibit the growth of rice seedlings. Germinolides ranging from 1 to 10 μM could alleviate the stress of 200 mM NaCl to some extent, but the promoting effect of 5 μM germinolides was the most significant. Germinolides ranging from 5 to 10 μM could also alleviate the stress of 50 mM Na2CO3 to some extent, and the concentration of 5 μM germinolides is the optimal concentration for promoting the growth of rice seedlings under saline-alkali stress.

Claims

1. The use of a germinolide to promote the growth of rice under saline stress conditions, characterized in that The application of sprout lactone in promoting the growth of rice under saline-alkali stress conditions, 0.1~10 μmol / L sprout lactone is used for promoting the growth of rice under saline-alkali stress conditions; wherein the application method of sprout lactone in promoting the growth of rice under saline-alkali stress conditions is carried out according to the following steps: I. Pretreatment of rice seeds: the rice seeds are fully suspended and soaked in 75% ethanol for 10 min, and then fully suspended and soaked in 0.02% mercuric chloride for 10 min. After taking out the rice seeds, they are repeatedly rinsed with distilled water for 10 times. Then add distilled water to cover the rice seeds and place them in a constant temperature box for 24 h to obtain the treated rice seeds; II. The treated rice seeds in step I are placed in a 10 cm diameter culture dish with filter paper at the bottom. 2 ml of sprout lactone with a concentration of 0.1~10 μmol / L is added to the culture dish for cultivation. The sprout lactone is replaced every 2 days, and the replacement frequency is 1~6 times, which realizes the application of sprout lactone.

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