Application of ferulic acid to promote rice seedling growth and increase yield of rice in saline-alkali soil

Abstract

The application of ferulic acid in promoting the growth of rice seedlings and increasing the yield of rice in saline-alkali land, the application of ferulic acid in rice is disclosed.The application provides a new application method of ferulic acid, i.e.the application of 5-20 micromole / L ferulic acid in promoting the growth of rice seedlings and increasing the yield of rice in saline-alkali land.The application of ferulic acid in promoting the growth of rice roots, the development of rice seedlings and increasing the salt tolerance of rice in fields, and the positive effect of ferulic acid on the growth and development of rice through the application of low-concentration ferulic acid are illustrated, which breaks the cognition of most people that ferulic acid inhibits the growth and development of plants to a certain extent.Especially, the application of ferulic acid in increasing the alkali tolerance of rice, and the application of low-concentration (5-20 micromole / L) ferulic acid enhances the alkali tolerance of rice and increases the yield of rice.

Description

Technical Field

[0001] This invention relates to the application of ferulic acid in rice. Background Technology

[0002] Ferulic acid (FA) and its derivatives are important intermediates in the phenylpropanoid metabolic pathway and are intermediate products in lignin biosynthesis. Ferulic acid is a typical water-soluble allelochemical with a wide range of effects on plant growth and development. It is a crucial metabolic intermediate for studying the role of phenylpropanoid metabolism in regulating plant growth and development.

[0003] Previous reports have largely focused on the inhibitory effects of ferulic acid as an allelochemical on plant growth and development when applied in large quantities. However, the bioactivity of ferulic acid under salt-alkali stress, a key bioactive substance involved in the phenylpropanoid metabolic pathway, has not yet been reported. Therefore, studying the effects of low concentrations of ferulic acid on root growth and seedling development under salt-alkali stress is of significant biological importance for exploring the bioactivity of ferulic acid and revealing its mechanism of regulating plant growth and development. Furthermore, it can provide valuable insights for the exogenous application of ferulic acid in production to regulate plant growth and development and increase crop yield. Summary of the Invention

[0004] This invention provides an application of ferulic acid in promoting rice seedling growth and increasing rice yield in saline-alkali land.

[0005] This invention relates to the application of ferulic acid in promoting rice seedling growth and increasing rice yield in saline-alkali land, specifically the application of 5–20 μmol / L ferulic acid in promoting rice seedling growth and increasing rice yield in saline-alkali land.

[0006] Furthermore, the application of ferulic acid in promoting rice seedling growth and increasing rice yield in saline-alkali land is achieved by soaking rice seeds in the early stage.

[0007] This invention utilizes ferulic acid as a plant growth regulator. It protects indoleacetic acid from decarboxylation and tissues from oxidative stress, promoting stem growth and root elongation. Low concentrations (5–20 μmol / L) of ferulic acid increase the mitotic ratio of root tip cells, promoting taproot elongation in rice. Under alkaline stress, exogenous application of ferulic acid promotes rice seedling development and improves seedling alkali tolerance. Exogenous application of ferulic acid can also promote tillering in saline-alkali soil rice, increase plant height and dry weight, and increase rice yield. This invention achieves better results with lower concentrations, significantly reducing production costs.

[0008] This invention relates to the application of ferulic acid in promoting rice root growth, seedling development, and improving salt tolerance in the field. The application of low-concentration ferulic acid demonstrates its positive effect on rice growth and development, challenging the common perception that ferulic acid inhibits plant growth and development. In particular, this invention demonstrates the effect of ferulic acid in improving rice's alkali tolerance, and the application of low-concentration (5–20 μmol / L) ferulic acid enhanced rice's alkali tolerance and increased rice yield.

[0009] This invention can lay a good foundation for promoting the growth of rice seedlings and increasing rice yield in saline-alkali land in agricultural production, and provide an effective method for improving rice through genetic engineering. Attached Figure Description

[0010] Figure 1 This is a comparison diagram of the root length of rice seeds after growth in Example 1;

[0011] Figure 2 for Figure 1 The following is a statistical chart of rice root length;

[0012] Figure 3 This is a diagram showing the development of rice seedlings in Example 3;

[0013] Figure 4 express Figure 3 A statistical chart of the aboveground length of rice seedlings;

[0014] Figure 5 This is a schematic diagram of the root dipping treatment group in Example 3 for 24 hours;

[0015] Figure 6 This is a graph showing the effects of different concentrations of ferulic acid root soaking on the number of tillers, plant height, and biomass of rice in field crops, as described in Example 3. Detailed Implementation

[0016] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0017] It should be noted that, unless otherwise specified, the embodiments and features described in the present invention can be combined with each other.

[0018] Specific implementation method one: The application of 5-20 μmol / L ferulic acid in promoting rice seedling growth and increasing rice yield in saline-alkali land.

[0019] Specific Implementation Method Two: This implementation method differs from Specific Implementation Method One in that the application of ferulic acid is carried out according to the following steps:

[0020] I. Rice seed pretreatment: Soak rice seeds in 70% ethanol for 10 minutes, remove the ethanol, then soak them in 0.5% sodium hypochlorite for 10 minutes, remove the sodium hypochlorite, rinse them repeatedly with distilled water 8-10 times, then add distilled water and place them in a 30℃ constant temperature incubator for 24-48 hours.

[0021] 2. Take out the rice seeds treated in step one and place them in a tray. Spray them with distilled water, cover them with plastic wrap, poke holes in the plastic wrap and spray them with distilled water. Place them in a 28℃ constant temperature incubator for 20-28 hours to promote germination.

[0022] Third, the germinated rice seeds from step two are sown onto a multi-well plate. The multi-well plate is then suspended in a container containing 5–20 μmol / L ferulic acid and placed in an artificial climate chamber. It is then cultured for 6–8 days under conditions of 25°C for 12 hours of light followed by 20°C for 12 hours of darkness. This achieves the application of ferulic acid. Other parameters are the same as in specific implementation method one.

[0023] In step one of this embodiment, the rice seeds are selected as mature seeds with plump grains; the rice seeds are fully suspended during soaking in ethanol and sodium hypochlorite; and the added distilled water is used to completely immerse the rice seeds in the distilled water.

[0024] Example 1: Promoting rice seedling growth using ferulic acid of the present invention

[0025] Five experimental groups were set up: the group without ferulic acid (0 μmol / L), the groups with ferulic acid at concentrations of 5 μmol / L, 10 μmol / L and 20 μmol / L respectively, and the control group (CK) with an equal amount of dimethyl sulfoxide (DMSO). Each group was treated 4 times in duplicate, and the dosage of each experimental group was the same.

[0026] The specific steps are as follows:

[0027] 1. Fully suspend and soak mature rice seeds in 70% ethanol for 10 minutes, remove the ethanol, then suspend and soak in 0.5% sodium hypochlorite for 10 minutes, remove the supernatant (sodium hypochlorite), rinse repeatedly with distilled water 8-10 times, then fill with distilled water and place in a 30℃ constant temperature incubator for 24-48 hours.

[0028] 2. Take out the mature rice seeds processed in step one and place them in a tray. Spray them with distilled water, cover them with plastic wrap, poke holes in the plastic wrap and spray them with distilled water. Place them in a 28℃ constant temperature incubator for 24 hours to promote germination.

[0029] 3. Divide the rice seeds that germinated uniformly in step 2 into five equal parts and sow them into a multi-well plate. The multi-well plate is suspended in plastic cups containing different experimental groups as described in this embodiment (the experimental group without ferulic acid, the groups with ferulic acid at concentrations of 5 μmol / L, 10 μmol / L and 20 μmol / L respectively, and the control group with an equal amount of dimethyl sulfoxide added), and place them in an artificial climate chamber (25℃ light for 12h / 20℃ darkness for 12h) for 7 days to obtain seedlings.

[0030] Example 1: Germinated rice seeds were photographed after being treated with different concentrations of ferulic acid for 7 days. The results are as follows. Figure 1 and Figure 2 As shown, Figure 1 This is a comparison chart of root lengths of rice seeds, showing the root lengths of germinating rice seeds after treatment with 0, 5, 10, and 20 μmol / L ferulic acid for 7 days. Scale bar: 1 cm. Figure 2 for Figure 1 The graph shown is a statistical chart of rice root length. This represents the control group (CK). This indicates the 5 μmol / L ferulic acid group (FA5). This indicates the 10 μmol / L ferulic acid group (FA10). Figure 2 The data shown are the mean ± standard deviation of four replicates. Significant differences (p<0.05, two-way ANOVA for LSD multiple comparison test) are indicated by different lowercase letters.

[0031] from Figure 1 and Figure 2 It can be seen that low concentrations (1–20 μmol / L) of ferulic acid increased the mitotic ratio of root tip cells and promoted the elongation of rice taproots, with the most significant changes in rice root length observed after treatment with 5 μmol / L and 10 μmol / L ferulic acid.

[0032] Example 2: Utilizing the application of the present invention to detect the effect of ferulic acid on promoting rice seedling development under alkaline stress.

[0033] Experimental groups were set up as follows: blank control (CK), ferulic acid treatment (FA), alkali treatment (A), and ferulic acid + alkali treatment (ferulic acid and Na2CO3 volume ratio of 1:1, A+FA). Each treatment was repeated 4 times. The control was supplemented with an equal amount of dimethyl sulfoxide (DMSO). The amounts of each experimental group were the same. The concentration of alkali (Na2CO3) in the treatment group was 5 mmol / L, and the concentration of ferulic acid was 5 μmol / L.

[0034] The specific method is as follows:

[0035] 1. Fully suspend and soak mature rice seeds in 70% ethanol for 10 minutes, remove the ethanol, then suspend and soak in 0.5% sodium hypochlorite for 10 minutes, remove the supernatant (sodium hypochlorite), rinse repeatedly with distilled water 8-10 times, then fill with distilled water and place in a 30℃ constant temperature incubator for 24-48 hours.

[0036] 2. Take out the mature rice seeds processed in step one and place them in a tray. Spray them with distilled water, cover them with plastic wrap, poke holes in the plastic wrap and spray them with distilled water. Place them in a 28℃ constant temperature incubator for 24 hours to promote germination.

[0037] 3. The 24 rice seeds that germinated uniformly in step 2 were sown into a multi-well plate, which was suspended in a plastic cup containing the experimental group solution and placed in an artificial climate chamber (25℃ light for 12h / 20℃ darkness for 12h) for 7 days.

[0038] Fourth, transplant the rice seedlings cultured in step three into 1 / 4 IRRI nutrient solution and continue to culture them in an artificial climate chamber for 7 days. Then, perform different treatments and observe the seedling growth after 7 days; this completes the application of ferulic acid.

[0039] The experimental groups described in step three of this embodiment are blank control treatment (CK), ferulic acid treatment (FA), alkali treatment (A), and ferulic acid + alkali treatment (the volume ratio of ferulic acid to Na2CO3 is 1:1, A+FA).

[0040] Example 2: Rice seedlings cultured for 14 days were treated with CK, FA, A, and A+FA for 7 days, and then photographed. The results are as follows. Figure 3 As shown. Among them, Figure 3 This is a graph showing the development of rice seedlings, specifically the seedling development after 14 days of growth, following treatments of CK, FA, A, and A+FA for 7 days. Scale bar: 2cm. Figure 4 express Figure 3 A statistical graph showing the aboveground length of rice seedlings. The data shown are the mean ± standard deviation of four replicates. Significant differences (p < 0.05, two-way ANOVA with LSD multiple comparison test) are indicated by different lowercase letters.

[0041] from Figure 3 and Figure 4 It can be seen that the development of rice seedlings was significantly damaged after alkali treatment, while ferulic acid treatment partially restored the damage to rice seedling development caused by alkali, indicating that ferulic acid can improve the alkali tolerance of rice seedlings.

[0042] Example 3: Effects of ferulic acid of the present invention on increasing rice yield in saline-alkali land

[0043] In this embodiment, three treatment groups were set up for field planting. The rice variety was "Dongdao 122". The specific groups were: seedlings cultivated in the control treatment (CK), seedlings cultivated in the 10 μmol / L ferulic acid treatment group (FAL) in Example 1, and seedlings cultivated in the 20 μmol / L ferulic acid treatment group (FAH) in Example 1. The control group was supplemented with an equal amount of dimethyl sulfoxide (DMSO).

[0044] In 2023, a field experiment was conducted at the Da'an Alkali Land Ecological Experimental Station in Jilin Province. Each experimental plot was 20 m², with four replicates per treatment, arranged in a randomized block design. At the jointing stage, three representative rice hills were selected from each plot to measure tiller number, plant height, and dry weight. At maturity, 100 rice hills were harvested from each plot at ground level. After air-drying, rice yield was measured, and six representative rice hills were selected. After air-drying, the yield components were measured. The yield composition is shown in Table 1. Figure 5 As shown in Table 1. Regarding fertilization, the nitrogen fertilizer application rate was 15 kg N / mu, and the phosphorus and potassium fertilizer application rates were both 6 kg P2O5 and K2O / mu. The nitrogen, phosphorus, and potassium fertilizers were sourced from urea (N 46%), superphosphate (P2O5 52%), and potassium chloride (K2O 60%), respectively. Specifically, 50% of the nitrogen fertilizer was used as basal fertilizer, 25% as tillering fertilizer, and 25% as heading fertilizer; all of the phosphorus fertilizer was used as basal fertilizer, and 70% of the potassium fertilizer was used as basal fertilizer and 30% as heading fertilizer. The soil was slightly saline-alkali land with a salt content of 0.2% and a pH value of 8.3-8.5.

[0045] Example 3: Rice seedlings were treated with root dips of CK, 10 μmol / L (FAL), and 20 μmol / L (FAH) for 24 hours, respectively, before being transplanted into saline-alkali land. Rice samples were taken at the jointing stage and harvest. Figure 5 The diagram shows the root dipping treatments for 24 hours, indicating that rice seedlings were treated with CK, 10 μmol / L (FAL), and 20 μmol / L (FAH) root dipping for 24 hours respectively. Figure 6 The graph shows the effects of different concentrations of ferulic acid root dipping on tiller number, plant height, and biomass in field rice. This represents sampling of rice at the jointing stage in the field, with tiller number, plant height, and plant dry weight measured. Data shown are the mean ± standard deviation of three replicates. Significant differences (p < 0.05, two-way ANOVA using Tukey's multiple comparison test) are indicated by different lowercase letters. Table 1 shows the yield, panicle number, grain number per panicle, seed setting rate, and thousand-grain weight of rice after root dipping with CK, 10 μmol / L (FAL), and 20 μmol / L (FAH) ferulic acid. Data shown are the mean of three replicates. Significant differences (p < 0.05, two-way ANOVA using LSD multiple comparison test) are indicated by different lowercase letters.

[0046] Table 1. Composition of Rice Yield

[0047]

[0048] from Figure 6 As can be seen, compared with the control (CK), root dipping with 20 μmol / L ferulic acid significantly promoted rice tillering, increased plant height, and plant dry weight. Results on yield and yield components showed that, compared with the control (CK), root dipping with 10 μmol / L ferulic acid did not significantly differ in the number of panicles and grains per panicle, while root dipping with 20 μmol / L ferulic acid significantly increased the number of panicles, but the number of grains per panicle did not change significantly (Table 1). Furthermore, neither concentration of ferulic acid root dipping significantly changed the seed setting rate or thousand-grain weight compared to the control (Table 1). Therefore, ferulic acid can promote the development of roots and aboveground parts of seedlings, ultimately increasing the number of tillers and yield of rice in saline-alkali land.

[0049] The results of Examples 1, 2 and 3 show that ferulic acid can promote rice seedling development, improve rice's salt and alkali tolerance, and further increase yield.

Claims

1. The application of ferulic acid in promoting rice seedling growth and increasing rice yield in saline-alkali land, characterized in that, The application of 5 ~ 20 μmol / L ferulic acid in promoting rice seedling growth and increasing rice yield in saline-alkali land is carried out according to the following steps: I. Rice seed pretreatment: Soak rice seeds in 70% ethanol for 10 min, remove the ethanol, then soak in 0.5% sodium hypochlorite for 10 min, remove the sodium hypochlorite, rinse repeatedly with distilled water 8-10 times, then add distilled water and place in a 30°C constant temperature incubator for 24-48 h.

2. Take out the rice seeds treated in step one and place them in a tray. Spray them with distilled water, cover them with plastic wrap, poke holes in the plastic wrap and spray them with distilled water. Place them in a 28°C constant temperature incubator for 20-28 hours to promote germination. Third, the rice seeds germinated in step two are sown onto a multi-well plate, the multi-well plate is suspended in a container containing 5 ~ 20 μmol / L ferulic acid, and placed in an artificial climate chamber for 6 ~ 8 days under conditions of 25°C light for 12 h and 20°C darkness for 12 h; thus, the application of ferulic acid is realized.

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