Hydroxyproline preparation for improving iron deficiency tolerance of plants
By applying hydroxyproline preparations to plants, the problem of plant iron deficiency is solved, the tolerance and growth ability of plants are improved, and the cost and environmental impact are reduced.
Patent Information
- Application Number
- CN202510840926.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-23
- Publication Date
- 2025-09-19
AI Technical Summary
Existing technologies for solving plant iron deficiency problems are inefficient, costly, and have negative impacts on the environment, and are unable to effectively improve plant tolerance to iron deficiency stress.
A hydroxyproline aqueous solution with a mass volume concentration of 0.65 mg/L and a hydroxyproline preparation with a pH value of 5.8 is applied to the soil or plant roots to enhance the tolerance of plants to iron deficiency stress.
It significantly increases the fresh weight and chlorophyll content of plants under iron deficiency stress, enhances iron reductase activity, alleviates oxidative stress, protects cell structure, reduces the risk of heavy metal pollution, and is low-cost and environmentally friendly.
Smart Images
Figure CN120660699A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the fields of plant physiology and agricultural biotechnology, specifically a method for improving plant tolerance to iron deficiency stress and increasing plant biomass through exogenous application of a hydroxyproline (HYP) preparation. This method is suitable for crop cultivation, horticultural plant production, and ecological restoration projects in iron-deficient soil environments. Background Art
[0002] Iron (Fe) is an important trace nutrient in plant physiological processes. It plays a core role in key processes such as photosynthesis, respiration, and stress response. It is a key factor in plant chlorophyll synthesis, mitochondrial respiratory chain electron transport, and oxidoreductase activity. Although iron is abundant in the soil, it tends to form insoluble oxides in alkaline soils with a pH greater than 7, making it impossible for plants to absorb it. For example, calcareous soils or areas with long-term hard water irrigation are prone to iron deficiency. At the same time, in waterlogged or poorly ventilated soils, iron exists in the form of trivalent iron, which is difficult to absorb, rather than the divalent iron (Fe) preferred by plants. ); In addition, excessive phosphorus, zinc, manganese or heavy metals (such as copper) may be applied during fertilization, which will compete with iron for absorption sites and inhibit the plant's absorption of iron.
[0003] Iron is a cofactor for key enzymes in chlorophyll synthesis. Iron deficiency causes leaf chlorosis, including yellowing of new leaves and yellowing and whitening between veins. This can also lead to damage to chloroplast structure, decreased photosynthesis, and reduced efficiency of light reactions, impacting energy metabolism and carbon fixation. Iron is a component of cytochromes, catalase, and other enzymes, and iron deficiency impairs respiration and antioxidant capacity. Furthermore, iron deficiency can lead to poor root development, stunted plants, reduced flowers and fruits, and, in severe cases, even plant death. Iron deficiency in crops can cause serious agricultural problems, manifesting as chlorosis, stunted growth, and reduced yields. To alleviate iron deficiency, plants have evolved adaptive strategies, including the transcriptional activation of key iron acquisition genes in Arabidopsis, including those encoding iron transporters and iron reductases, to promote the reduction of trivalent iron and the absorption of divalent iron by the plant.
[0004] Currently, there are several main approaches to addressing plant iron deficiency: First, applying traditional iron fertilizers (such as Fe-EDTA). However, this method can quickly oxidize and become ineffective in alkaline soils, requiring large amounts of application. Excessive amounts can cause soil acidification or heavy metal accumulation. Second, soil improvement, such as applying sulfur, acid conditioners, and organic fertilizers to adjust soil pH, requires continuous monitoring, is slow to take effect, requires long-term application, and has limited effectiveness for acute iron deficiency. Because existing technologies are limited by efficiency, cost, and environmental costs, they cannot meet the demands for precision, sustainability, and adaptability. Therefore, more applicable methods are needed to address these issues.
[0005] Hydroxyproline is commonly found in the extracellular matrix structural proteins of animal cells, such as collagen. In animals, hydroxyproline enhances the stability of collagen, which serves as a structural protein in human tissues. Hydroxyproline can maintain the optimal state of collagen, enhance bone toughness, and reduce wear of articular cartilage. Hydroxyproline can also reduce the damage to collagen caused by oxidative free radicals, slowing the aging process of cells. In plants, hydroxyproline is primarily found in glycoproteins in the cell wall. Hydroxyproline is primarily a key component of the cell wall, but research on its function in plants is limited, particularly its role in enhancing plant tolerance to iron deficiency stress. Summary of the Invention
[0006] In order to solve the problem that plant iron deficiency is limited by efficiency, cost and environmental costs, the present invention provides a hydroxyproline preparation for improving plant iron deficiency tolerance.
[0007] A hydroxyproline preparation for improving plant tolerance to iron deficiency is a hydroxyproline aqueous solution with a mass volume concentration of 0.65 mg / L and a pH value of 5.8; The hydroxyproline preparation is used to improve the tolerance of plants to iron deficiency stress; When used, the hydroxyproline preparation is poured into the soil or poured onto the roots of plants.
[0008] Further technical solutions are as follows: The improvement of plant tolerance to iron deficiency is to increase the fresh weight and chlorophyll content of the plant under iron deficiency stress.
[0009] When used, the concentration of the hydroxyproline preparation poured into the soil is 0.65 mg / L, and the amount of the hydroxyproline preparation poured into the plant roots is 0.1-0.2 L / plant.
[0010] The beneficial technical effects of the present invention are as follows:
[0011] See also Figure 1 The present invention uses metabolomics to analyze the differences in metabolites in plants under normal conditions and iron deficiency conditions. Figure 2 , first discovered that plants produce more hydroxyproline under iron deficiency stress, see Figure 3 Furthermore, phenotypic analysis was used to demonstrate that the addition of exogenous hydroxyproline preparations could improve the tolerance of plants to iron deficiency stress. Figure 5 Hydroxyproline preparations can stabilize the synthesis of chlorophyll under iron deficiency stress. As a component of cell wall glycoprotein, hydroxyproline may indirectly protect chloroplast function by maintaining cell wall stability, thereby significantly increasing chlorophyll content. Figure 6 Hydroxyproline preparations can directly enhance the activity of iron reductase, thereby improving the effectiveness of iron and thus increasing the absorption of iron by plants. Figure 7 and Figure 8 , hydroxyproline preparations can alleviate the oxidative stress caused by iron deficiency stress, which can lead to the accumulation of reactive oxygen species and thus oxidative damage, while hydroxyproline preparations can significantly reduce the hydrogen peroxide caused by iron deficiency stress and protect the integrity of cell membranes and chloroplast structures. The present invention can not only enhance the absorption of iron by plants, but also stabilize the reduction of chlorophyll caused by iron deficiency stress, alleviate the oxidative damage caused by iron deficiency stress, rather than just using iron fertilizers as a traditional method to increase soil iron content. The present invention is suitable for crop cultivation under iron-deficient soil environments, has the advantages of low cost, environmental friendliness, and ease of operation, can replace traditional iron fertilizers and reduce the risk of heavy metal pollution in agricultural production. The present invention provides a theoretical basis for breeding plants for resistance to iron deficiency stress and improving the viability and biomass of plants under iron deficiency stress. BRIEF DESCRIPTION OF THE DRAWINGS
[0012] Figure 1 Metabolomics analysis of the changes in hydroxyproline content in normal and iron-deficiency-stressed Arabidopsis.
[0013] Figure 2 Figure 2 shows that iron deficiency stress leads to an increase in hydroxyproline content in Arabidopsis thaliana.
[0014] Figure 3 A diagram showing the effect of adding hydroxyproline preparations on the growth of Arabidopsis thaliana cultured in a culture medium under iron deficiency stress.
[0015] Figure 4 This figure shows the effect of adding hydroxyproline preparation on the fresh weight of Arabidopsis thaliana cultured in culture medium under iron deficiency stress.
[0016] Figure 5 This figure shows the effect of adding hydroxyproline preparation on the chlorophyll content of Arabidopsis thaliana cultured in culture medium under iron deficiency stress.
[0017] Figure 6 This figure shows the effect of adding hydroxyproline preparation on the iron reductase activity of Arabidopsis thaliana cultured in culture medium under iron deficiency stress.
[0018] Figure 7 This figure shows the effect of adding hydroxyproline preparation on the reactive oxygen species of Arabidopsis thaliana cultured in culture medium under iron deficiency stress.
[0019] Figure 8 A diagram showing the effect of adding hydroxyproline preparations on hydrogen peroxide in Arabidopsis thaliana cultured in a culture medium under iron deficiency stress.
[0020] Figure 9 The figure shows the effect of adding hydroxyproline preparation on the fresh weight of Arabidopsis thaliana cultured in soil under iron deficiency stress.
[0021] Figure 10The figure shows the effect of adding hydroxyproline preparation on the chlorophyll content of Arabidopsis thaliana cultured in soil under iron deficiency stress. DETAILED DESCRIPTION
[0022] The present invention will be further described below by way of embodiments with reference to the accompanying drawings. Example 1
[0023] The experiment of using hydroxyproline preparation to enhance plant tolerance to iron deficiency stress is as follows: 1. Metabolomics was used to analyze the differences in metabolites between Arabidopsis thaliana grown on iron-containing MS medium and iron-deficient MS medium. Figure 1 After differential comparison, it was found that the hydroxyproline content in Arabidopsis thaliana grown on iron-deficient MS medium was significantly higher than that in Arabidopsis thaliana grown on iron-containing MS medium. Figure 2 To further confirm this result, the hydroxyproline content in Arabidopsis thaliana grown on iron-containing MS medium and iron-deficient MS medium was directly detected by chromatography. The results showed that iron deficiency did increase the hydroxyproline content in Arabidopsis thaliana.
[0024] 2. To analyze the effects of hydroxyproline preparations on the growth of Arabidopsis thaliana under iron deficiency stress, wild-type Arabidopsis thaliana were sown in 90 mm diameter culture dishes containing MS medium, MS medium + 25 μL of 0.65 g / L hydroxyproline preparation, iron-deficient MS medium, and iron-deficient MS medium + 25 μL of 0.65 g / L hydroxyproline preparation, respectively. The dishes were then placed in a 22°C constant temperature and light incubator (with a photoperiod of 16 hours light and 8 hours dark) and cultured vertically. Figure 3 After seven days, no significant differences were observed between Arabidopsis plants grown in MS medium and MS medium supplemented with 25 μL of a 0.65 g / L hydroxyproline preparation, indicating that exogenous addition of hydroxyproline had no significant effect on Arabidopsis growth. Arabidopsis plants grown on iron-deficient MS medium grew significantly weaker than those grown in MS medium and MS medium supplemented with 25 μL of a 0.65 g / L hydroxyproline preparation, indicating that iron deficiency significantly inhibited their growth. However, Arabidopsis plants grown in the iron-deficient MS medium supplemented with hydroxyproline significantly outperformed those grown in iron-deficient MS medium.
[0025] 3. See Figure 4The fresh weight of Arabidopsis thaliana sown in MS medium, MS medium + 25 μL of 0.65 g / L hydroxyproline preparation, iron-deficient MS medium, and iron-deficient MS medium + 25 μL of 0.65 g / L hydroxyproline preparation was analyzed. The results showed that there was no significant difference in the fresh weight of Arabidopsis thaliana sown in MS medium and MS medium + 25 μL of 0.65 g / L hydroxyproline preparation groups. The fresh weight of Arabidopsis thaliana sown in iron-deficient MS medium was significantly lower than that sown in MS medium and MS medium + 25 μL of 0.65 g / L hydroxyproline preparation groups, while the fresh weight of Arabidopsis thaliana sown in iron-deficient MS medium + 25 μL of 0.65 g / L hydroxyproline preparation group was significantly higher than that grown on iron-deficient MS medium.
[0026] 4. See Figure 5 The chlorophyll content of Arabidopsis thaliana sown in the MS medium group, MS medium + 25 μL 0.65 g / L hydroxyproline preparation group, iron-deficient MS medium group, and iron-deficient MS medium + 25 μL 0.65 g / L hydroxyproline preparation group was analyzed. The results showed that there was no significant difference in chlorophyll content between Arabidopsis thaliana sown in the MS medium group and the MS medium + 25 μL 0.65 g / L hydroxyproline preparation group. The chlorophyll content of Arabidopsis thaliana sown in the iron-deficient MS medium group was significantly lower than that of Arabidopsis thaliana sown in the MS medium group and the MS medium + 25 μL 0.65 g / L hydroxyproline preparation group. The chlorophyll content of Arabidopsis thaliana sown in the iron-deficient MS medium + 25 μL 0.65 g / L hydroxyproline preparation group was significantly higher than that of Arabidopsis thaliana grown in the iron-deficient MS medium group.
[0027] 5. See Figure 6 The iron reductase activity of Arabidopsis thaliana sown in the MS medium group, MS medium + 25 μL 0.65 g / L hydroxyproline preparation group, iron-deficient MS medium group, and iron-deficient MS medium + 25 μL 0.65 g / L hydroxyproline preparation group was analyzed. The results showed that there was no significant difference in the iron reductase activity of Arabidopsis thaliana sown in the MS medium group and the MS medium + 25 μL 0.65 g / L hydroxyproline preparation group. The iron reductase activity of Arabidopsis thaliana sown in the iron-deficient MS medium group was significantly lower than that of Arabidopsis thaliana sown in the MS medium group and the MS medium + 25 μL 0.65 g / L hydroxyproline preparation group, while the iron reductase activity of Arabidopsis thaliana sown in the iron-deficient MS medium + 25 μL 0.65 g / L hydroxyproline preparation group was significantly higher than that of Arabidopsis thaliana grown in the iron-deficient MS medium group.
[0028] 6. See Figure 7 The reactive oxygen levels of Arabidopsis thaliana sown in the MS medium group, the MS medium + 25 μL 0.65 g / L hydroxyproline preparation group, the iron-deficient MS medium group, and the iron-deficient MS medium + 25 μL 0.65 g / L hydroxyproline preparation group were analyzed. The results showed that there was no significant difference in the reactive oxygen levels of Arabidopsis thaliana sown in the MS medium group and the MS medium + 25 μL 0.65 g / L hydroxyproline preparation group. The reactive oxygen level of Arabidopsis thaliana sown in the iron-deficient MS medium group was significantly lower than that of Arabidopsis thaliana sown in the MS medium group and the MS medium + 25 μL 0.65 g / L hydroxyproline preparation group. The reactive oxygen level of Arabidopsis thaliana sown in the iron-deficient MS medium + 25 μL 0.65 g / L hydroxyproline preparation group was significantly higher than that of Arabidopsis thaliana grown in the iron-deficient MS medium group.
[0029] 7. See Figure 8 The hydrogen peroxide levels of Arabidopsis thaliana sown in the MS medium group, the MS medium + 25 μL 0.65 g / L hydroxyproline preparation group, the iron-deficient MS medium group, and the iron-deficient MS medium + 25 μL 0.65 g / L hydroxyproline preparation group were analyzed. The results showed that there was no significant difference in the hydrogen peroxide levels of Arabidopsis thaliana sown in the MS medium group and the MS medium + 25 μL 0.65 g / L hydroxyproline preparation group. The hydrogen peroxide level of Arabidopsis thaliana sown in the iron-deficient MS medium group was significantly lower than that of Arabidopsis thaliana sown in the MS medium group and the MS medium + 25 μL 0.65 g / L hydroxyproline preparation group. The hydrogen peroxide level of Arabidopsis thaliana sown in the iron-deficient MS medium + 25 μL 0.65 g / L hydroxyproline preparation group was significantly higher than that of Arabidopsis thaliana grown in the iron-deficient MS medium group.
[0030] The above results indicate that hydroxyproline preparations can enhance plant tolerance to iron deficiency stress. Example 2
[0031] The following experiments demonstrated the ability of hydroxyproline preparations to enhance plant growth in iron-deficient soils: 1. Collect alkaline soil with a pH of about 7.2 and normal soil with a pH of about 5.8, weigh 1 kg of each and place them in round flower pots with a diameter of 20 cm and a height of 30 cm. Use Hoagland nutrient solution to soak the soil. Prepare 10 pots for each group.
[0032] 2. Germinate the Arabidopsis seeds one week in advance and transplant 5 Arabidopsis seedlings of the same growth potential into each pot.
[0033] 3. The normal soil with a pH of approximately 5.8 was divided into two groups: a normal group and a normal + hydroxyproline preparation group. The alkaline soil with a pH of approximately 7.2 was divided into two groups: an iron-deficient group and an iron-deficient + hydroxyproline preparation group. The normal group was irrigated with 0.2 L of distilled water per week, the normal + hydroxyproline preparation group was irrigated with 0.2 L of hydroxyproline preparation per week, the iron-deficient group was irrigated with 0.2 L of distilled water per week, and the iron-deficient + hydroxyproline preparation group was irrigated with 0.2 L of hydroxyproline preparation per week.
[0034] 4. See Figure 9 After 3 weeks of growth, the fresh weight of Arabidopsis thaliana in each group was statistically analyzed. The results showed that there was no significant difference in the fresh weight of Arabidopsis thaliana grown in the normal group and the normal + hydroxyproline preparation group. The fresh weight of Arabidopsis thaliana grown in the iron-deficient group was significantly lower than that sown in the normal group and the normal + hydroxyproline preparation group, while the fresh weight of Arabidopsis thaliana sown in the iron-deficient + hydroxyproline preparation group was significantly higher than that grown in the iron-deficient group.
[0035] 5. See Figure 10 After 3 weeks of growth, the chlorophyll content of Arabidopsis thaliana in each group was statistically analyzed. The results showed that there was no significant difference in chlorophyll content between the Arabidopsis thaliana grown in the normal group and the normal + hydroxyproline preparation group. The chlorophyll content of Arabidopsis thaliana grown in the iron-deficient group was significantly lower than that of the Arabidopsis thaliana sown in the normal group and the normal + hydroxyproline preparation group, while the chlorophyll content of Arabidopsis thaliana sown in the iron-deficient + hydroxyproline preparation group was significantly higher than that of the Arabidopsis thaliana grown in the iron-deficient group.
[0036] The above results indicate that the hydroxyproline preparation can improve the ability of plants to grow in iron-deficient soil.
[0037] It will be easily understood by those skilled in the art that the above embodiments are merely preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.
Claims
1. A hydroxyproline preparation for improving plant tolerance to iron deficiency, characterized in that: The hydroxyproline preparation is a hydroxyproline aqueous solution with a mass volume concentration of 0.65 mg / L and a pH value of 5.8; the hydroxyproline preparation is used to improve the tolerance of plants to iron deficiency stress; When used, the hydroxyproline preparation is poured into the soil or poured onto the roots of plants.
2. The hydroxyproline preparation for improving plant iron deficiency tolerance according to claim 1, characterized in that: The improvement of plant tolerance to iron deficiency is to increase the fresh weight and chlorophyll content of the plant under iron deficiency stress.
3. The hydroxyproline preparation for improving plant tolerance to iron deficiency according to claim 1, characterized in that: When used, the concentration of the hydroxyproline preparation poured into the soil is 0.65 mg / L, and the amount of the hydroxyproline preparation poured into the plant roots is 0.1-0.2 L / plant.