Application of molybdenum-regulated synergistic substance phloridzin in promoting crop growth and activating soil phosphorus

Abstract

The application provides application of a molybdenum-regulated synergistic substance phlorizin in promoting crop growth and activating soil phosphorus, and systematically studies the influence of exogenous application of phlorizin on soybean growth and on the form and content of phosphorus in soil. Through comparison of the difference between the experimental group and the control group, the role of phlorizin in promoting soybean growth and promoting the transformation of soil phosphorus is revealed, and meanwhile, the application of phlorizin significantly promotes the growth of soybean, which is specifically embodied in the increase of soybean plant height, the increase of the dry matter weight of the aboveground part and the root part and the like. Therefore, the application not only provides a brand-new growth promotion strategy for soybean production, but also opens an effective way for improving the utilization rate of soil phosphorus and improving the overall quality of soil.

Description

Technical Field

[0001] This invention relates to the field of agricultural planting, specifically to the application of phlorizin, a molybdenum-regulated synergist, in promoting crop growth and activating soil phosphorus. Background Technology

[0002] Phosphorus is an essential nutrient for plant growth, playing a crucial role in plant development and yield formation. However, most phosphorus in the soil exists in forms that are difficult for plants to directly absorb and utilize, resulting in low phosphorus utilization efficiency. To improve phosphorus utilization efficiency, researchers have been exploring various strategies, including using exogenous substances to enhance plants' ability to absorb and utilize phosphorus.

[0003] Flavonoids, as naturally occurring plant secondary metabolites, play a crucial role in the activation of soil phosphorus. Through metal chelation, flavonoids can convert insoluble phosphorus in the soil into forms available to plants. Furthermore, as part of root exudates, flavonoids not only promote the release of nutrients and increase their availability but also indirectly promote phosphorus activation by altering the community structure of rhizosphere microorganisms and reducing organic matter consumption. These mechanisms collectively reveal the key role of flavonoids in soil phosphorus cycling and plant nutrient uptake. Recent studies have shown that phosphorus-deficient environments induce plant roots to secrete more flavonoid compounds. For example, under phosphorus-deficient conditions, the accumulation of flavonoid compounds in the roots of white lupin promotes root cluster formation through exogenous flavonoid addition, alleviating soil phosphorus deficiency. Further research has found that molybdenum application can significantly regulate metabolite changes in soybean rhizosphere soil and affect the form transformation of soil phosphorus, improving phosphorus availability; one important metabolite is phlorizin.

[0004] Phlorizin is a natural flavonoid compound with various biological activities, and in recent years it has been found to have positive effects on plant growth. However, the application of phlorizin in soybean growth and phosphorus absorption has not yet been studied. Summary of the Invention

[0005] To address the aforementioned problems, this invention provides an application of phlorizin, a molybdenum-regulated synergist, in promoting crop growth and activating soil phosphorus. The study investigates the effects of exogenous application of phlorizin on soybean growth, as well as on the form and content of phosphorus in the soil. By comparing the differences between the experimental and control groups, the role of phlorizin in promoting soybean growth and soil phosphorus transformation is revealed.

[0006] The application of phlorizin, a molybdenum-regulated synergist, in promoting crop growth and activating soil phosphorus, wherein the synergist comprises phlorizin and is used for the following purposes:

[0007] Increase the aboveground dry matter, root dry matter, and plant height of crops;

[0008] Increase the phosphorus content and phosphorus accumulation in the aboveground parts and roots.

[0009] The soil phosphorus activator is used to increase the content of unstable inorganic phosphorus, unstable organic phosphorus, and moderately stable organic phosphorus in the soil.

[0010] The crop in question is soybean.

[0011] The synergistic substance is a phlorizin-containing compound, and the concentration of phlorizin in the solution is 1–10 μmol / L.

[0012] Preferably, the concentration of phlorizin in the solution is 5 μmol / L or 10 μmol / L.

[0013] The solution contains phosphorus and molybdenum nutrients.

[0014] The molybdenum nutrient is (NH4)6Mo7O. 24 ·4H2O.

[0015] The concentration of the molybdenum nutrient in the solution is 0.02 μM.

[0016] This invention provides an application of phlorizin in the preparation of a soybean growth promoter, and studies the effects of exogenous application of phlorizin on soybean growth, as well as on the form and content of phosphorus in the soil. By comparing the differences between the experimental group and the control group, the effects of phlorizin on promoting soybean growth and promoting the transformation of phosphorus in the soil are revealed. Attached Figure Description

[0017] Figure 1 The effect of molybdenum application on phosphorus absorption and translocation in soybeans under different phosphorus levels;

[0018] Figure 2 The number of metabolites that are significantly altered by Mo under control treatment;

[0019] Figure 3 The number of metabolites that show significant changes mediated by Mo under P treatment;

[0020] Figure 4 Fifty differentially metabolites mediated by Mo under control treatment;

[0021] Figure 5 Ninety-four differential metabolites mediated by Mo under P treatment;

[0022] Figure 6 Flavonoids whose abundance varies significantly under different phosphorus levels mediated by molybdenum application;

[0023] Figure 7 Phenotypic images of the effects of different concentrations of phlorizin on soybean growth;

[0024] Figure 8 The effects of different concentrations of phlorizin treatment on soybean plant height, aboveground dry matter and root dry matter under hydroponic conditions were investigated.

[0025] Figure 9 The effects of phlorizin and Mo treatments on available phosphorus content in soil;

[0026] Figure 10 The effects of phlorizin and Mo treatment on soybean plant height, aboveground dry matter and root dry matter under soil experimental conditions;

[0027] Figure 11 The effects of phlorizin and Mo treatments on phosphorus content and phosphorus accumulation in the aboveground and root parts of soybean under soil experimental conditions. Detailed Implementation

[0028] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to specific embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and do not limit the scope of protection of the invention.

[0029] Example 1: Effect of molybdenum application on phosphorus uptake in soybeans

[0030] The experiment included two phosphorus addition levels: 0.015 g / kg (to ensure normal early crop growth), denoted as Control; and 0.15 g / kg, denoted as P. Two molybdenum addition levels were also included: 0 mg / kg, denoted as -Mo; and 0.15 mg / kg, denoted as +Mo. There were four treatments in total: Control-Mo, Control+Mo, P-Mo, and P+Mo, with four replicates for each treatment, for a total of 16 pots. The tested soybean variety was Tianlong No. 1. Soil was simultaneously placed in the inner and outer chambers of the root box and compacted. Soybean seeds were sown in the inner chamber for growth. Each pot, including the root box, contained 8.0 kg of soil.

[0031] like Figure 1 As shown in the comparative analysis, at the same molybdenum addition level, phosphorus treatment significantly increased the aboveground phosphorus content, phosphorus translocation coefficient, aboveground phosphorus accumulation, root phosphorus accumulation, and overall phosphorus accumulation in soybeans, but had no significant effect on root phosphorus content. On the other hand, at the same phosphorus level, compared with no molybdenum treatment, molybdenum treatment not only significantly increased the aboveground phosphorus content, phosphorus translocation coefficient, and aboveground and whole-plant phosphorus accumulation in soybeans, but also significantly reduced the root phosphorus content and accumulation at both phosphorus levels. This indicates that molybdenum application can effectively improve the phosphorus absorption efficiency of soybeans at different phosphorus levels and promote the translocation of phosphorus from roots to aboveground parts, thereby increasing aboveground phosphorus accumulation and reducing root phosphorus accumulation.

[0032] Example 2: Effects of molybdenum application on soybean rhizosphere metabolites

[0033] Since molybdenum application can significantly increase the available phosphorus content in the soil, metabolomics methods can be used to systematically screen substances whose metabolite abundance in soybean roots changes significantly under molybdenum application treatment.

[0034] Figures 2-5 The results showed that, through detailed analysis in both positive and negative ion modes, we identified 584 metabolites. Figure 2 and Figure 4 In the control treatment with phosphorus addition, molybdenum application had a significant effect on metabolites. The abundance of 19 metabolites was increased after molybdenum application, while the abundance of 31 metabolites was decreased. The abundance of the remaining 534 metabolites did not change significantly. Figure 3 and Figure 5 Under P treatment conditions, among the metabolites affected by molybdenum application, the abundance of 33 metabolites significantly increased, while the abundance of 61 metabolites significantly decreased. Further KEGG pathway enrichment analysis revealed that under P+Mo treatment, differentially enriched metabolites were mainly concentrated in four key metabolic pathways: flavonoid biosynthesis, phenylpropanoid biosynthesis, caprolactam degradation, and carbohydrate digestion and absorption.

[0035] Figure 6 In the study, under the phosphorus supplementation level P, the enrichment of differential metabolites was more widespread, significantly distributed across 20 metabolic pathways. Particularly, pathways involving ABC transporters, galactose metabolism, cysteine ​​and methionine metabolism, flavonoid biosynthesis, thermogenesis, carbohydrate digestion and absorption, mineral absorption, and EGFR tyrosine kinase inhibitor resistance showed significant enrichment. Given that differential metabolites were significantly enriched in the flavonoid biosynthesis pathway at both phosphorus levels, we conducted an in-depth analysis of the flavonoids showing differential changes under these two phosphorus levels. Under P treatment, the addition of molybdenum significantly increased the abundance of Vignafulan, phlorizin, neoisalloside, hesperidin 5-O-glucoside, and catechin 7-glucoside. Under molybdenum treatment, the abundance of one flavonoid, phlorizin, showed a significant difference, as shown in the following figures. Figure 6 As shown, this substance is also more easily absorbed and utilized by organisms and has higher safety, being non-toxic or low-toxic to the human body.

[0036] Example 3: Determining the concentration of phlorizin

[0037] The growth-promoting effect of phlorizin (Pz) on soybeans was tested. The nutrient solution culture experiment was conducted in the glass greenhouse of the College of Resources and Environment, Huazhong Agricultural University. The growing conditions were set at 25℃, with 14 hours of light. Plastic boxes (30.3×20.5×8.5cm) were used as culture containers. Both the plastic boxes and foam board lids were opaque, and each container contained 3L of nutrient solution. The nutrient solution formula was: 4mM Ca(NO3)2·4H2O, 1mM NH4NO3, 1mM KH2PO4, 5mM KNO3, 2mM MgSO4·7H2O, 9.55μM MnSO4·4H2O, 46.28μM H3BO3, 0.77μM ZnSO4·7H2O, 0.32μM CuSO4·5H2O, 0.02μM (NH4)6Mo7O 24 • 4H2O, 98.09μM EDTA-Fe, change the nutrient solution every 4 days.

[0038] Phlorizin was dissolved in 0.5 mL of 80% methanol and added to the nutrient solution. The concentrations of phlorizin in the nutrient solution were set at 0 μmol / L, 0.5 μmol / L, 1 μmol / L, 2 μmol / L, 5 μmol / L, and 10 μmol / L. The control treatment received the same amount of 80% methanol, and each treatment was repeated three times. Soybeans were harvested after 30 days of growth, and the dry matter and phosphorus content of the plants were measured. Intact, plump soybean seeds were selected, disinfected by soaking in a 2% sodium hypochlorite solution for 15 minutes, rinsed with ultrapure water, and soaked for 3 hours to ensure full water absorption and exposure of the embryo. The seeds were then evenly sown into vermiculite for seedling cultivation. Four days later, when the seedlings reached approximately 5 cm in height and the cotyledons were unfolded, they were transplanted. Six soybean seedlings of uniform growth were selected and planted in each pot, using a sponge to hold the soybeans in place to prevent slippage. Soybeans were harvested after 30 days of growth.

[0039] The following parameters were determined: (1) Plant height: The height of the main stem of soybean was measured with a tape measure at harvest; (2) Dry weight: The samples of each part of soybean were blanched at 105℃ for 30 min and dried at 60℃ to constant weight. The dry weight was recorded; (3) Photosynthetic parameters: Net photosynthetic rate, stomatal conductance, intercellular CO2 concentration and transpiration rate were measured using a portable photosynthesis instrument (LICOR-6400XT) at 8-11 am on a sunny day; (4) Phosphorus content of plants: 0.1500g of plant sample was placed in a 50ml digestion tube, 5ml of concentrated sulfuric acid was added, and the sample was left to stand overnight. The sample was then preheated at 160℃ for 10-15 minutes in an infrared digestion oven. The temperature was then increased to 280℃. The sample was removed and cooled slightly. A small amount of 30% H2O2 was added and the process was repeated several times until the digestion liquid became clear. After making up the volume and filtering, the phosphorus content was determined using a flow analyzer.

[0040] like Figure 7-8As shown in the comparative analysis, soybean growth phenotypes were best under Pz concentrations of 5 μmol / L and 10 μmol / L, significantly superior to other concentrations. Specifically, the aboveground dry matter, root dry matter, and plant height were significantly higher in the 5 μmol / L and 10 μmol / L Pz treatments than in other treatments, but there was no significant difference between the 5 μmol / L and 10 μmol / L Pz treatments. Therefore, Pz concentrations of 5 μmol / kg and 10 μmol / kg achieved the best growth-promoting effect on soybeans. To further control costs, this study used 5 μmol / kg phlorizin for the next verification experiment.

[0041] Example 4: Phlorizin

[0042] Step 1: Experimental Treatment Setup

[0043] Control group (CK): only ultrapure water was added; Pz: only 5 μmol / kg phlorizin was added.

[0044] Step 2: Adding fertilizer

[0045] Add the following levels of fertilizer to each test pot:

[0046] Nitrogen: added in the form of urea ((NH2)2CO) at a level of 0.15 g / kg; Phosphorus: added in the form of potassium dihydrogen phosphate (KH2PO4) at a level of 0.15 g / kg; Potassium: added in the form of potassium chloride (KCl) at a level of 0.20 g / kg.

[0047] Step 3: Addition of Mo and Phlorizin

[0048] Mo to (NH4)6Mo7O 24 • 4H₂O was added to the soil as a molybdenum source; and phlorizin was dissolved in 80% methanol and then added to the soil of the experimental group. The control group received the same amount of 80% methanol, but no phlorizin was added.

[0049] Step 4: Treatment and sowing of soybean seeds

[0050] Select soybean seeds with intact seed coats and plump kernels. Disinfect them by soaking them in a 2% sodium hypochlorite solution for 15 minutes, then rinse them thoroughly with ultrapure water. Next, soak the seeds for 3 hours to allow them to fully absorb water and expose the embryo. Afterward, sow the seeds evenly in vermiculite for seedling cultivation.

[0051] Step 5: Transplanting and Growing Conditions

[0052] Four days after seedling establishment, when the soybean seedlings reached approximately 5 cm in height and their cotyledons had unfolded, they were transplanted. One seedling per pot was selected based on uniform growth, its roots were thoroughly cleaned, and the transplanted seedlings were kept uniform in size. The experiment was conducted in a glass greenhouse at the College of Resources and Environment, Huazhong Agricultural University, with growth conditions set at 25℃ and 14 hours of light per day.

[0053] Step 6: Sample determination.

[0054] Soybeans were harvested 45 days after growth, and then measurements were taken. The results are shown in Table 1 below. Figure 9-11 As shown, compared with the control group (CK), the Pz treatment significantly increased the contents of unstable inorganic phosphorus, unstable organic phosphorus, and moderately stable organic phosphorus in the soil, while significantly decreasing the content of stable phosphorus. Meanwhile, the content of moderately stable inorganic phosphorus remained stable. This result indicates that phlorizin can effectively promote the conversion of rhizosphere phosphorus from a stable to an unstable state, thereby facilitating phosphorus absorption and utilization by crops.

[0055] Table 1 Effects of phlorizin treatment on phosphorus speciation in soybean rhizosphere soil

[0056]

[0057] like Figure 9 The results showed that, compared with the control group (CK), exogenous phlorizin significantly increased the content of available phosphorus in the soil, with an increase of 13.01%. This data clearly indicates that phlorizin treatment helps promote the release of available phosphorus in the soil or slows down its fixation and transformation process, thereby effectively improving the utilization rate of available phosphorus in the soil. This result further confirms that the rhizosphere metabolite phlorizin regulated by Mo has the ability to activate available phosphorus in the soil, thereby promoting the absorption and accumulation of phosphorus by soybeans, and ultimately having a positive impact on soybean growth.

[0058] Figure 10 The results showed that, compared with the control group (CK), soybeans treated with phlorizin exhibited significant improvements in plant height, aboveground dry matter, and root dry matter. Specifically, phlorizin treatment increased plant height by 13.92%, aboveground dry matter by 7.98%, and root dry matter by 11.67%.

[0059] Depend on Figure 11The results showed that soybean plants treated with phlorizin exhibited significant increases in phosphorus content and accumulation in both the aboveground and root parts. Specifically, compared to the control treatment, phlorizin treatment increased aboveground phosphorus content by 24.21% and phosphorus accumulation by 34.95%; simultaneously, root phosphorus content and accumulation increased by 19.61% and 29.09%, respectively. These results demonstrate the significant positive role of phlorizin in promoting phosphorus absorption in soybeans. This finding is of great importance for utilizing phlorizin to improve phosphorus utilization and promote soybean growth.

[0060] This invention provides an application of molybdenum-regulated soybean rhizosphere metabolite phlorizin in promoting soil phosphorus activation. The study systematically investigates the effects of exogenous phlorizin application on soybean growth, on phosphorus speciation and content in the soil, and on the effect of molybdenum application on phlorizin in soybean roots. By comparing the differences between the experimental and control groups, the role of phlorizin in promoting soybean growth and soil phosphorus transformation is revealed.

[0061] Please note that the above embodiments are for illustrative purposes only and do not constitute a limitation on the scope of protection. Various equivalent substitutions and modifications can be made to the above embodiments without departing from the spirit and essence of this method.

[0062] The above embodiments are for illustrating the implementation schemes disclosed in this invention and should not be construed as limiting the invention. Furthermore, various modifications and variations of the methods listed herein will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been specifically described in conjunction with various specific preferred embodiments, it should be understood that the invention should not be limited to these specific embodiments. In fact, various modifications as described above that are obvious to those skilled in the art to obtain the invention should be included within the scope of this invention.

Claims

1. The application of phlorizin, a molybdenum-regulated synergist, in promoting crop growth and activating soil phosphorus, characterized in that, The synergist or soil phosphorus activator contains phlorizin, and the synergist is used for the following purposes: Increase the aboveground dry matter, root dry matter, and plant height of crops; Increase the phosphorus content and phosphorus accumulation in the aboveground parts and roots; The soil phosphorus activator is used for the following purposes: increasing the content of unstable inorganic phosphorus, unstable organic phosphorus, and moderately stable organic phosphorus in the soil; The crop is soybean, and the synergistic substance is a solution containing phlorizin, with a concentration of phlorizin in the solution of 1~10 μmol / L. The solution contains phosphorus and molybdenum nutrients.

2. The application according to claim 1, characterized in that, The concentration of phlorizin in the solution is 5 μmol / L or 10 μmol / L.

3. The application according to claim 1, characterized in that, The molybdenum nutrient is (NH4)6Mo7O. 24 ·4H2O.

4. The application according to claim 3, characterized in that, The concentration of the molybdenum nutrient in the solution is 0.02 μM.

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