A plant growth regulator composition comprising benzylaminopurine and thidiazuron and its application

Abstract

The present invention relates to a plant growth regulator composition comprising benzylaminopurine and thidiazuron and its application. The plant growth regulator composition comprises a compound of the active ingredients benzylaminopurine and thidiazuron. The mass ratio of benzylaminopurine to thidiazuron is (40-50):5. Application of the plant growth regulator composition to wheat significantly increases wheat yield, and benzylaminopurine and thidiazuron exhibit a significant synergistic effect in increasing wheat yield. The plant growth regulator composition can also promote wheat seed germination and seedling growth under nutritional stress, as well as wheat seed and seedling growth under HgCl2 stress.

Description

Technical Field

[0001] The present invention belongs to the technical field of plant growth control and regulation, and specifically relates to a plant growth regulator composition comprising benzylaminopurine and thidiazuron and an application thereof. The plant growth regulator composition can promote wheat seed germination and seedling growth under nutritional stress, as well as wheat seed and seedling growth under HgCl2 stress. Background Art

[0002] Wheat is one of China's most important staple foods. It is cultivated in nearly every agricultural region, from south to north, from plains to mountainous areas. Wheat is second only to rice in both cultivated area and total production, making it the most important staple food in China and crucial for both food security and the well-being of its residents.

[0003] Seed germination and seedling growth are critical stages in plant growth. The seed germination process includes the imbibition stage, the germination stage, the germination stage, and the seedling stage. These stages broadly involve the physical process of water absorption and expansion, the physiological and biochemical processes of nutrient transformation, and the molecular biological process of embryo germination. In wheat, a monocot, the embryo plays a leading role in seed germination. By sensing and responding to changes in environmental factors such as light, temperature, nutrients, and water, it releases signals to the endosperm, inducing the degradation of stored substances in the endosperm and promoting the flow of nutrients to the embryo. During seed germination, the endosperm acts as a mechanical barrier to protect the embryo while providing nutrients for its development. The endosperm also has the ability to sense environmental changes and regulate embryo growth by producing and secreting hormones and photosensitive pigments.

[0004] The endosperm, the primary nutrient reservoir for seeds, contains a variety of nutrients, including protein, fat, starch, minerals, and terpenes. These storage substances fully guarantee the smooth germination and growth of seeds. During seed germination, starch and soluble sugars provide energy for seed germination and embryo growth. If the soluble sugar content within the seed decreases, the seed germination rate, germination rate, and germination potential will also decrease. Protein, as one of the important components of organisms, plays a vital role in plant seed germination and seedling growth. Studies have found that protein content is positively correlated with seed germination and seedling growth rate. The faster the protein degradation rate, the higher the germination rate or germination rate.

[0005] Successful seed germination and the normal establishment of seedlings are crucial factors in plant reproduction and are of great economic and ecological significance. During this period of heterotrophic growth, embryos and seedlings are often subject to stress caused by various factors, both environmental and internal. In practice, insect bites and mechanical sowing can damage seeds, eliminate endosperm, and form incomplete seeds, leading to a reduction in stored material.

[0006] It can be speculated that seeds with damaged endosperm, i.e., nutrient depletion, will fail to grow or have their growth severely inhibited. However, this view has not been experimentally verified. Furthermore, in studies of the effects of various stress factors on seed germination and seedling growth, the issue of seed storage material depletion has rarely been discussed. There are also no reports on the effects of a combined plant growth regulator of benzylaminopurine and thidiazuron on wheat seed germination and seedling growth under nutrient stress. Therefore, this study set up experiments with nutrient depletion, i.e., removal of only the endosperm, only the roots, and both the endosperm and roots. The growth and development of wheat seedlings after seed nutrient depletion was examined to analyze the effects of a combined plant growth regulator of benzylaminopurine and thidiazuron on wheat seed germination and seedling growth under nutrient depletion.

[0007] Heavy metal pollution has become an important aspect of soil pollution, among which mercury is one of the most polluting heavy metal elements. In the soil, mercury is easily converted into methylmercury, which has stronger accumulation, fat solubility and high neurotoxicity than mercury. It can directly threaten human health through biological accumulation and food chain. At present, the control of mercury pollution has always been a difficult problem and hot spot in environmental protection research. Wheat is an important food crop in my country, and it is more obviously poisoned by heavy metal mercury in soil contaminated areas. Studies have shown that low concentrations of Hg 2+ (<0.1mmol / L) can promote the germination of wheat seeds, but too high a concentration can inhibit the germination of wheat seeds and the growth of seedlings. 2+ When the concentration reached 0.5mmol / L, the germination rate, root length, sprout length and fresh weight of wheat seeds decreased, and the activity of antioxidant enzymes decreased, and the decrease increased with the increase of mercury ion concentration.

[0008] To this end, based on preliminary preliminary tests, this application selected wheat varieties with strong adaptability but different resistance as test materials, and analyzed the effects of seed soaking with a compound plant growth regulator of benzylaminopurine and thiadiazole on seed germination, seedling growth and changes in physiological indicators of different wheat varieties under HgCl2 stress at different concentrations, in order to clarify the physiological mechanism of the compound plant growth regulator of benzylaminopurine and thiadiazole in alleviating HgCl2 stress in wheat, and provide a theoretical basis for its rational application in future production. Summary of the Invention

[0009] The present invention aims to overcome the defects of the prior art and provide a plant growth regulator composition comprising benzylaminopurine and thidiazuron, which can be used for wheat growth regulation to achieve improvement in wheat quality and yield.

[0010] The present invention also provides the use of the above-mentioned plant growth regulator composition in promoting wheat seed germination and seedling growth under nutritional stress. It is used to regulate wheat growth nutritional stress and can achieve the effects of improving wheat growth rate, plant height, root length, SPAD, total leaf area and dry weight, increasing soluble protein and proline content. The accumulated malondialdehyde can convert starch stored in the plant body into glucose that is beneficial to growth, ultimately promoting rapid growth of the wheat root system and aboveground parts, keeping them consistent with normally growing plants, thereby reducing the damage of nutritional stress to wheat seedlings.

[0011] The present invention also provides the use of the above-mentioned plant growth regulator composition in promoting the growth of wheat seeds and seedlings under HgCl2 stress. The plant growth regulator composition is used for regulating heavy metal mercury stress in wheat growth, can achieve the improvement of wheat seed germination potential and germination rate, increase soluble protein content, induce the production of SOD and POD antioxidant enzymes in seedlings, timely eliminate ROS, reduce MDA content, alleviate oxidative damage, and thus reduce the damage of HgCl2 to wheat seedlings.

[0012] To achieve the above object, the present invention adopts the following technical solutions:

[0013] A plant growth regulator composition comprising benzylaminopurine and thidiazuron, wherein the plant growth regulator composition is composed of a compound of the active ingredients benzylaminopurine and thidiazuron.

[0014] Specifically, the mass ratio of benzylaminopurine to thidiazuron can be (40-50):5.

[0015] Furthermore, the mass ratio of benzylaminopurine to thiadiazole can be (43-47):5, preferably 45:5.

[0016] Furthermore, the plant growth regulator composition is made of active ingredients and pesticide adjuvants into a dosage form allowed in pesticides. The dosage form can be wettable powder, soluble powder, soluble liquid, water-dispersible granules, suspension, emulsifiable concentrate, water emulsion, microemulsion or microcapsule, etc. The preferred dosage form is suspension.

[0017] The plant growth regulator composition described above may include a pesticide adjuvant, which may be solid or liquid and may be selected from, for example, one or more of a carrier, solvent, dispersant, wetting agent, adhesive, thickener, binder, surfactant, and fertilizer. The plant growth regulator composition of the present invention may be applied solely containing the active ingredient or mixed with an additive (pesticide adjuvant).

[0018] Furthermore, the total content of the active ingredients accounts for 0.3-0.7% of the total weight of the plant growth regulator composition, preferably 0.5%.

[0019] The present invention also provides the application of the above-mentioned plant growth regulator composition in regulating wheat growth, which can effectively increase the effective number of wheat ears, the number of grains per ear and its thousand-grain weight, improve agronomic traits, and can also significantly increase wheat yield, improve wheat quality, and effectively improve the economic benefits of wheat in the later stage. Specifically, when applied, the plant growth regulator composition is applied during the wheat growth period, preferably, during the wheat tillering period and the heading period. The amount of the plant growth regulator composition applied is 0.32-0.35g / mu based on the sum of the mass of benzylaminopurine and thidiazuron. The plant growth regulator composition can be diluted with water to a concentration of 4.8-5.2g / hectare based on the total mass of benzylaminopurine and thidiazuron, and sprayed onto the aboveground stems and leaves of wheat during the wheat tillering period and the heading period.

[0020] The present invention also provides the use of the plant growth regulator composition for promoting wheat seed germination and seedling growth under nutritional stress. Specifically, the plant growth regulator composition is used to alleviate wheat nutritional stress by applying the plant growth regulator composition to wheat seeds under severe nutritional stress. Specifically, during application, the plant growth regulator composition can be diluted with water and then used to soak the wheat seeds, effectively improving the wheat's growth rate, plant height, root length, chlorophyll content (SPAD), total leaf area, and dry weight.

[0021] The present invention also provides the use of the plant growth regulator composition for promoting the growth of wheat seeds and seedlings under HgCl2 stress. Specifically, the plant growth regulator composition is used to alleviate heavy metal mercury stress in wheat by applying the plant growth regulator composition to wheat seeds stressed by heavy metal mercury. Specifically, during application, the plant growth regulator composition can be diluted with water and then used to soak the wheat seeds, effectively improving the germination potential, germination rate, and soluble protein content of the wheat.

[0022] Benzylaminopurine is the first synthetic cytokinin that can increase the germination potential and germination rate of plant seeds, promote seed emergence, and help plants maintain stable photosynthesis under adverse conditions. It also enhances the plant's ability to scavenge free radicals and maintain cell membrane stability, thereby improving the plant's stress resistance and alleviating the stress of various adverse conditions on seed germination and seedling growth. Thidiazuron can promote fruit enlargement, delay plant aging, enhance stress resistance, promote plant photosynthesis, increase crop yields, and improve product quality.

[0023] Compared with the prior art, the advantages and beneficial effects of the present invention are as follows:

[0024] The present invention provides a plant growth regulator composition comprising benzylaminopurine and thidiazuron. When applied to wheat, the composition significantly increases wheat yield. Furthermore, benzylaminopurine and thidiazuron exhibit a significant synergistic effect in increasing wheat yield. The plant growth regulator composition can also promote wheat seed germination and seedling growth under nutritional stress, as well as wheat seed and seedling growth under HgCl2 stress.

[0025] When the plant growth regulator composition of the present invention is used to alleviate nutritional stress in wheat, the plant growth regulator composition can be diluted with water and then used to soak wheat seeds, which can effectively improve the growth rate, plant height, root length, SPAD, total leaf area and dry weight of wheat; it can increase the content of soluble protein and proline, and the accumulated malondialdehyde can convert the starch stored in the plant into glucose that is beneficial to growth, ultimately promoting the rapid growth of the wheat root system and aboveground parts, so that they are consistent with normally growing plants, thereby reducing the damage caused by nutritional stress to wheat seedlings.

[0026] When the plant growth regulator composition of the present invention is used to alleviate heavy metal mercury stress in wheat, the plant growth regulator composition can be diluted with water and then used to soak wheat seeds, which can effectively improve the germination potential, germination rate and soluble protein content of wheat; it can significantly induce the production of SOD and POD antioxidant enzymes in wheat seedlings, timely eliminate ROS, reduce MDA content, alleviate oxidative damage, and thus reduce the damage of HgCl2 to wheat seedlings. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 The effect of nutrient deficiency on the soluble protein content of wheat seedlings;

[0028] Figure 2 The effect of nutrient deficiency on proline content in wheat seedlings;

[0029] Figure 3 The effect of nutrient deficiency on the malondialdehyde content in wheat seedlings;

[0030] Figure 4 To study the effect of applying pesticides to soak seeds on the germination rate of wheat seeds under different concentrations of HgCl2 stress;

[0031] Figure 5 The effect of soaking seeds without applying pesticide on the germination rate of wheat seeds under different concentrations of HgCl2 stress;

[0032] Figure 6 To study the effect of applying pesticides to soak seeds on the germination potential of wheat seeds under different concentrations of HgCl2 stress;

[0033] Figure 7The effect of soaking seeds without applying pesticide on the germination potential of wheat seeds under different concentrations of HgCl2 stress;

[0034] Figure 8 To study the effect of seed soaking with pesticides on the root length of wheat seedlings under different concentrations of HgCl2 stress;

[0035] Figure 9 The effect of seed soaking without applying pesticide on the root length of wheat seedlings under different concentrations of HgCl2 stress;

[0036] Figure 10 To study the effect of seed soaking with pesticides on the sprout length of wheat seedlings under different concentrations of HgCl2 stress;

[0037] Figure 11 The effect of soaking seeds without applying pesticide on the sprout length of wheat seedlings under different concentrations of HgCl2 stress;

[0038] Figure 12 To study the effect of seed soaking with pesticides on the fresh weight of wheat seedlings under different concentrations of HgCl2 stress;

[0039] Figure 13 The effect of seed soaking without applying pesticide on the fresh weight of wheat seedlings under different concentrations of HgCl2 stress;

[0040] Figure 14 To study the effect of applying pesticides to soak seeds on the SOD content of wheat seedlings under different concentrations of HgCl2 stress;

[0041] Figure 15 The effect of seed soaking without applying pesticide on the SOD content of wheat seedlings under different concentrations of HgCl2 stress;

[0042] Figure 16 To study the effect of applying pesticides to soak seeds on the POD content of wheat seedlings under different concentrations of HgCl2 stress;

[0043] Figure 17 The effect of seed soaking without applying pesticide on POD content of wheat seedlings under different concentrations of HgCl2 stress;

[0044] Figure 18 To study the effect of applying pesticides to soak seeds on MDA of wheat seedlings under different concentrations of HgCl2 stress;

[0045] Figure 19 The effect of seed soaking without applying pesticide on MDA of wheat seedlings under different concentrations of HgCl2 stress;

[0046] Figure 20 To study the effect of seed soaking with pesticides on the soluble protein content of wheat seedlings under different concentrations of HgCl2 stress;

[0047] Figure 21This paper shows the effect of seed soaking without applying pesticides on the soluble protein content of wheat seedlings under different concentrations of HgCl2 stress. DETAILED DESCRIPTION

[0048] The technical solution of the present invention is further described in detail below in conjunction with the embodiments, but the protection scope of the present invention is not limited thereto.

[0049] The experimental operations involved in the wheat experiment, unless detailed, can be performed using conventional techniques in the art.

[0050] Application experiment 1: Effect of plant growth regulator combination on regulating wheat growth

[0051] 1. Indoor test - drug formulation screening test purpose: to obtain the appropriate concentration formula of benzylaminopurine + thiadiazole, and to show that the drug has the effect of promoting the growth of the aboveground part of wheat

[0052] 1 Test conditions

[0053] 1.1 Test Target

[0054] Wheat (Zhengmai 9023)

[0055] 1.2 Instruments and Equipment

[0056] Artificial climate incubator, electronic balance (sensitivity 0.1 mg), confidential pipettes (100-1000 μL, 20-100 μL, 1-20 μL), spray tower, beakers, measuring cylinders and several plastic pots with holes.

[0057] 2 Experimental design

[0058] 2.1 Experimental treatment

[0059] 2.1.1 Test material cultivation

[0060] Test wheat seeds were first flooded with 45°C warm water for 8 hours. Remove the seeds, shade them with dark gauze, and place them in a sponge-lined petri dish. Moisten the seed surface with water to accelerate germination. Once the seeds appear white, sow them. Fill the pot with test soil to a depth of 4 / 5, then infiltrate the soil from the bottom until it is completely moistened to saturation. Sow the pretreated wheat seeds evenly and evenly over the soil, cover with 1-3 cm of soil, and transfer them to an incubator for incubation. Culture conditions: Temperature range: 22-25°C; Humidity range: 40-95%; Light intensity: 0-25,000 LUX; CO2: 450-2000 ppM.

[0061] After wheat seedlings emerged, thinning was performed, and ten wheat plants of uniform growth were selected for each pot. Leaves were sprayed with a Potter spray tower. Pot dimensions: top diameter 27.8 cm, bottom diameter 19.8 cm, height 28 cm.

[0062] 2.1.2 Pharmacy

[0063] Table 1 Test drug information

[0064] Chinese name common name Dosage form and content Production Unit Benzylaminopurine 6-Benzylaminopurine 98% TC Zhejiang Dapeng Pharmaceutical Co., Ltd. Thidiazuron thidiazuron 98% TC Jiangsu Youjia Plant Protection Co., Ltd.

[0065] 2.2 Test methods

[0066] 2.2.1 Preparation of medicine

[0067] 0.1020 g of benzylaminopurine technical drug and 0.1020 g of thidiazuron technical drug were weighed, dissolved in 1 ml of NN dimethylformamide (DMF), and diluted with 0.1% Tween-80 aqueous solution to prepare mother solutions for later use.

[0068] The prepared benzylaminopurine stock solution was diluted separately to prepare five concentration gradients of 4.1 mg / L, 4.3 mg / L, 4.5 mg / L, 4.7 mg / L, and 4.9 mg / L.

[0069] The prepared thidiazuron mother solution was diluted separately to prepare five concentration gradients of 0.125 mg / L, 0.25 mg / L, 0.5 mg / L, 0.75 mg / L, and 1.0 mg / L.

[0070] The two agents were then combined according to their active ingredients as shown in the table below, for a total of 25 agent treatments. The agent treatments are shown in Table 2 below.

[0071] Table 2 Chemical treatment

[0072]

[0073]

[0074] 2.2.2 Method of drug administration

[0075] After calibrating the spray pressure (15 psi) and the spray volume on the Potter spray tower, stems and leaves were sprayed using a designed concentration gradient from low to high doses. Each treatment was replicated four times, with a blank control. Each pot was treated with 2 ml of solution. After treatment, the plants were transferred to an artificial climate incubator for incubation. Culture conditions included: temperature range: 22-25°C; humidity range: 40-95%; light intensity: 0-25,000 LUX; and carbon dioxide: 450-2000 ppM.

[0076] After treatment, the effects of the liquid on the quality and regulation of wheat in each group were observed and recorded regularly. After 14 days of treatment, the fresh weight of the aboveground part of wheat was recorded by survey method.

[0077] 2.3 Test indicators

[0078] Fresh weight of aboveground parts, fresh weight promotion rate, E-E0.

[0079] 3 Calculation formula

[0080] Calculation of fresh weight promotion rate: The promotion rate of fresh weight of the aboveground part of the test material was calculated according to formula (1).

[0081] R=(PT-CK) / CK×100 (1)

[0082] Where: R is the growth promotion rate, in percentage (%);

[0083] CK—fresh weight of the aboveground part of the plants in the control group, in grams (g);

[0084] Fresh weight of aboveground parts of plants in the PT-treatment group, in grams (g).

[0085] Calculation of the combined effect of mixtures: Using the Gowing method, the theoretical promotion rate of mixing agents A and B in the above proportions is calculated according to formula (2);

[0086] Eo = x+Y-XY / 100 (2)

[0087] Where: X—promotion rate of aboveground fresh weight when the dosage of regulator A is P;

[0088] Y—Promotion rate of aboveground fresh weight when the dosage of regulator B is 0:

[0089] Eo: The theoretical promotion rate when the dosage of regulator A is P + the theoretical promotion rate when the dosage of regulator B is 0;

[0090] E: The actual promotion rate after the regulator A and regulator B are mixed in the above proportions.

[0091] E-Eo>10% is a synergistic effect; E-Eo<-10% is an antagonistic effect; E-Eo values ​​between ±10% are additive effects.

[0092] 4 Data Processing

[0093] DPS 9.01 was used for 5% significance comparison, and Duncan's new multiple range method was used for statistical analysis of the experimental data.

[0094] 5 Results Analysis

[0095] In Tables 3 and 4, Examples 1-5 are single doses of benzylaminopurine at concentrations of 4.1, 4.3, 4.5, 4.7, and 4.9 mg / L, respectively; Examples 6-10 are single doses of thiadiazole at concentrations of 0.125, 0.25, 0.5, 0.75, and 1.0 mg / L, respectively; Examples 11-35 are benzylaminopurine and thiadiazole at different mass ratios; Example 36 is a blank control group to which benzylaminopurine and thiadiazole were not applied.

[0096] Table 3 Effects of the test agents on the fresh weight of the aboveground part of wheat

[0097]

[0098]

[0099] Note: Different letters in the table indicate significant differences (p<0.05), the same below.

[0100] As shown in Table 3, benzylaminopurine and thiadiazole have a promoting effect on the aboveground fresh weight of wheat. When the active ingredients are 4.3+0.5, 4.5+0.5, and 4.7+0.5, the aboveground fresh weight is 28.75g, 29.48g, and 28.60g, respectively, which are significantly higher than those treated with other agents. The actual promotion rates are 28.03%, 33.29%, and 28.03%, respectively, and there is no phytotoxicity to wheat growth.

[0101] Table 4 Effects of the test agents on the fresh weight of the aboveground part of wheat

[0102]

[0103] As can be seen from Table 4, in Examples 22-24, the control effect was ideal within the combination ranges of 4.3+0.5, 4.5+0.5, and 4.7+0.5 of the benzylaminopurine technical drug to the thidiazuron technical drug. The remaining treatments with the ratios of benzylaminopurine technical drug to the thidiazuron technical drug all had an additive effect. Therefore, the treatment with a ratio of 43 to 47:5 of benzylaminopurine technical drug to the thidiazuron technical drug was selected for further field trials to evaluate its effect on wheat quality and yield.

[0104] 2. Field efficacy test - regulating growth, increasing yield and income. Test purpose: to clarify the effect of applying benzylaminopurine + thiadiazole compound on increasing yield and income and improving quality of wheat. 1. Test conditions

[0105] 1.1 Test Target

[0106] Wheat (Zhengmai 9023)

[0107] 1.2 Experimental design

[0108] 1.2.1 Test agent, same as above

[0109] Table 5 Experimental design of test agents

[0110]

[0111] 1.3 Experimental plot arrangement

[0112] The experimental agent, control agent and blank agent were designed in randomized blocks. The experiment set up 6 treatments, each with 4 replicates, arranged in random blocks, with protection rows around them, marked as A, B, C, and D, with an area of ​​20m 2 .

[0113] 2. Application method

[0114] 2.1 Application period and frequency

[0115] Application time: Apply once at the tillering stage and the heading stage of wheat;

[0116] Application amount: 20ml;

[0117] Number of applications: 2 times.

[0118] 2.2 Pesticide application equipment

[0119] A T-HS16D backpack electric sprayer (Shandong Agricultural Machinery Co., Ltd.) was used, and the liquid flow rate calibrated before application was 1 L / min.

[0120] 3 Test indicators

[0121] According to the national grain moisture standards, the plot yield, number of effective ears and number of filled grains per ear, thousand-grain weight, protein content, wet gluten content, starch content, sedimentation value, etc. are measured.

[0122] 4 Experimental data processing method

[0123] DPS 9.01 was used for comparison with 5% significance level, and Duncan's method was used for statistical analysis of the experimental data.

[0124] 5 Test results

[0125] 5.1 Wheat yield indicators

[0126] Table 6 Effects of benzylaminopurine and thidiazuron on wheat yield

[0127]

[0128] Note: Different letters in the table indicate significant differences (p<0.05), the same below.

[0129] As can be seen in Table 6, in Examples 1-3, the ratio of benzylaminopurine technical drug to thiadiazole technical drug is within the combination range of 4.3+0.5, 4.5+0.5, and 4.7+0.5. Benzylaminopurine + thiadiazole (43-47:5) has a significant improvement on wheat yield indicators, can effectively increase the effective number of wheat ears, the number of grains per ear and its thousand-grain weight, and can effectively promote the increase in yield and weight of wheat. Among them, when benzylaminopurine + thiadiazole (45:5), the effective number of wheat ears is 83.33, the number of grains per ear is 37.13, and the thousand-grain weight is 46.68g, which are significantly higher than the relevant data of benzylaminopurine + thiadiazole (43:5) and benzylaminopurine + thiadiazole (47:5), which is the optimal ratio.

[0130] Table 7 Effect of benzylaminopurine + thiadiazole on wheat plot yield

[0131]

[0132]

[0133] Note: Different letters in the table indicate significant differences (p<0.05), the same below.

[0134] As shown in Table 7, benzylaminopurine + thiadiazole significantly improved the yield of wheat plots. When the ratio was 45:5, the average yield of the plot was the highest at 16.31 kg, an increase of 9.46% compared with CK, which was the optimal ratio.

[0135] 5.2 Wheat quality indicators

[0136] Table 8 Effects of benzylaminopurine and thidiazuron on wheat quality

[0137]

[0138] Note: Different letters in the table indicate significant differences (p<0.05), the same below.

[0139] As can be seen from Table 8, when the ratio of benzylaminopurine + thiadiazole is 43-47:5, its protein content, starch content, wet gluten content and sedimentation value are significantly better than those of the blank and single-agent treatment groups. When the ratio of benzylaminopurine + thiadiazole is 45:5, its protein content is 16.76g / 100g, starch content is 65.32%, wet gluten content is 37.71%, and sedimentation value is 58.74%, which is better than other treatments and is the optimal ratio. This shows that when the ratio of aminopurine + thiadiazole is 45:5, it can effectively improve the quality and economic benefits of wheat and achieve the effect of increasing production and income.

[0140] Application experiment 2: Effects of plant growth regulators on wheat seed germination and seedling growth under nutritional stress

[0141] 1. Experimental purpose: To determine whether plant growth regulators can effectively alleviate the inhibitory effects of nutrient stress on wheat seed germination and seedling physiology, and to study the growth potential of wheat seedlings and their adaptability to nutrient stress.

[0142] 2. Experimental Design

[0143] 1. Test materials and reagents

[0144] 1.1 Test materials

[0145] For Yunong 416, Yannong 19, and Xinong 585 (winter wheat varieties), seeds were sown in germination boxes at 25°C in the dark to ensure uniform germination conditions.

[0146] 1.2 Test agent, same as above

[0147] 2 Test methods

[0148] 2.1 Chemical treatment

[0149] Weigh 0.1020 g of benzylaminopurine technical and 0.1020 g of thidiazuron technical, respectively, dissolve them in 1 ml of N-dimethylformamide (DMF), and dilute them with 0.1% Tween-80 aqueous solution to prepare mother liquors for later use; dilute the prepared benzylaminopurine mother liquor to 4.5 mg / L, and dilute the prepared thidiazuron mother liquor to 0.5 mg / L.

[0150] Preparation of a 0.5% benzylaminopurine and thidiazuron mixture: 100 ml of 4.5 mg / L benzylaminopurine and 0.5 mg / L thidiazuron were mixed and used as a stock solution. The solution was diluted with a 0.1% Tween-80 aqueous solution to a concentration of 0.5% (mass percentage) of the active ingredient before application. Treatments were indicated by Y, with a 0.1% Tween-80 aqueous solution serving as the CK control.

[0151] 2.2 Seed soaking and germination

[0152] Soak the seeds in a 10% NaClO solution for 15 minutes for disinfection, then rinse 4-5 times with sterile water. Place the rinsed wheat seeds in a sealed conical flask, submerge them in sterile water, and soak them in the dark for 3 hours at a room temperature of approximately 25°C. Place the soaked seeds, ventral groove facing down, in a germination box lined with filter paper to germinate (germination standard is that the coleoptile is flush with the first true leaf), and ensure that no excess water flows out. The germination environment should be consistent with the soaking environment.

[0153] 3 Experimental process

[0154] 3.1 Test material processing

[0155] To investigate the adaptability of wheat seedlings to partial embryo loss at different germination times, this experiment designed seven treatments: endosperm removal alone (CE), endosperm removal plus pesticide treatment (CE+Y), primary root removal alone (CR), primary root removal plus pesticide treatment (CR+Y), both endosperm and primary root removal (CER), both endosperm and primary root removal plus pesticide treatment (CER+Y), and normal-growing wheat seedlings (CK). Endosperm removal was performed by inserting the tip of a sterilized No. 11 scalpel blade into the junction between the base of the coleoptile and the endosperm, once on each side. This caused the endosperm to fall off. Root removal involved removing the primary root from its base at the coleoptile junction, without damaging the endosperm. Simultaneous endosperm and primary root removal involved removing the primary root using the same scalpel blade after endosperm removal. The operating environment should be clean and sterile.

[0156] 3.2 Determination of the minimum “age” for wheat seedling survival after endosperm removal

[0157] To determine the minimum age at which wheat seedlings can survive after seed storage material depletion, seedlings at 0, 6, 12, 24, 30, 36, 42, 48, and 54 hours after germination were subjected to seven treatments: normal growth (CK), endosperm removal (CE), endosperm removal with a chemical treatment (CE+Y), primary root removal (CR), primary root removal with a chemical treatment (CR+Y), both endosperm and primary root removal (CER), and both endosperm and primary root removal with a chemical treatment (CER+Y). Each treatment was replicated three times. Treated wheat seedlings were planted in germination boxes (12 cm × 12 cm × 5 cm) filled with sea sand and incubated in a light incubator (25°C, 12 h light / 12 h dark). Watering was performed with 3 ml of Hoagland's nutrient solution every two days. Survival rates were calculated after 15 days of incubation.

[0158] 3.3 Growth and development of wheat seedlings after germination for 48 hours

[0159] Based on statistical survival rates, wheat seedlings 48 hours after germination were selected for normal growth, endosperm removal, primary root removal, and both endosperm and primary root removal experiments. Each treatment was planted in a germination box (12 cm × 12 cm × 5 cm) filled with sea sand, with 50 plants per box and 3 boxes per treatment. 3 ml of Hoagland nutrient solution was used for culture, and the incubator was set at 25°C with 12 hours of light and 12 hours of darkness. Ten plants with consistent growth were selected from each box for index measurement at the one-leaf extraction (initial endosperm supply), one-leaf-one-heart stage (mid-endosperm supply), and two-leaf-one-heart stage (end of endosperm supply). Each treatment was repeated 3 times.

[0160] 4. Index determination

[0161] 4.1 Basic index determination

[0162] Plant height was measured with a ruler, and root length was measured with a root scanner.

[0163] Calculation of leaf area: Leaf area (cm 2 ) = leaf length × leaf width × 0.85.

[0164] Calculation of growth rate: Taking the dry weight of wheat seedlings grown for 48 h as the initial value, the change in dry weight of wheat seedlings was calculated when they grew to one leaf out (FLC), one leaf and one heart (FLU), and two leaves and one heart (TLU) after n days under the four treatments, and then the growth rate of wheat seedlings was calculated.

[0165] Growth rate (mg / d) = dry weight change within nd (mg) / n.

[0166] Calculation of dry weight: After sampling, place the sample in an oven at 105℃ for 30 minutes, then turn it to 75℃ for drying for 1 day, and then measure the weight using a 1 / 1000 balance.

[0167] 4.2 Physiological index measurement

[0168] The soluble protein content was determined by Coomassie Brilliant Blue G-250 staining; the proline content was determined by ninhydrin colorimetry; the malondialdehyde content (MDA) was determined by thiobarbituric acid method; and the chlorophyll content (SPAD) value was determined using a portable chlorophyll meter, requiring the selected leaf position to be consistent.

[0169] 5 Data Processing

[0170] Excel was used for statistical analysis and plotting, and DPS 9.01 was used for significance analysis, with a significance level of P < 0.05.

[0171] 6 Test results

[0172] 6.1 Minimum survival time of wheat seedlings after endosperm removal

[0173] Table 9 Minimum survival time of wheat seedlings after endosperm removal

[0174]

[0175] Table 9 shows that compared with wheat seedlings (CK and CR) that received endosperm nutrition, CE and CER wheat seedlings failed to survive when their endosperm was removed prematurely (0, 6, 12, and 24 hours). However, wheat seedlings in the CE+Y treatment (treated with 0.5% benzylaminopurine and thiadipyrone) survived at 30 hours, with a survival rate of 69%. In the CER+Y treatment (treated with 0.5% benzylaminopurine and thiadipyrone), wheat seedlings survived at 12 hours, with a survival rate of 63%. The survival rate of wheat seedlings in CE after germination for 30 hours after endosperm removal was 37%, and the survival rate of wheat seedlings in CER after germination for 36 hours was 53%. This indicates that the combined plant growth regulator of 0.5% benzylaminopurine and thiadipyrone effectively improves the survival rate of wheat seedlings in treatments where both the endosperm and primary roots are removed.

[0176] 6.2 Effects of nutrient deficiency on leaf age and growth rate of wheat seedlings

[0177] Table 10 Effects of nutrient deficiency on leaf age and growth rate of wheat seedlings

[0178]

[0179] In Table 10, the time required for treated wheat seedlings to develop a single leaf was 2 days, while normal wheat seedlings only needed 1.5 days. Wheat seedlings with only their root system removed (CR) took 2.3 days to grow from FLC to FLU, 0.5 days slower than CK. Application of a combined plant growth regulator (0.5% benzylaminopurine and thiadiazole) effectively shortened this growth time and increased the growth rate to only 2.1 days. The leaf age change rate at the TLU stage was similar to that of CK. Wheat seedlings with endosperm removed (CE and CER) had the greatest impact on leaf age. After treatment, it took 4.7 and 9 days to reach FLU, and 10.1 and 19 days to reach TLU, respectively. Therefore, compared with wheat seedlings with endosperm removed (CR and CK), wheat seedlings with endosperm removed (CE and CER) required more time to reach a fixed leaf age during the seedling stage.

[0180] In summary, nutritional deficiency significantly affected the growth of wheat seedlings, showing varying degrees of growth inhibition at all growth stages. Especially when both the endosperm and root system were removed, the growth of wheat seedlings was most restricted. The application of a composite plant growth regulator of 0.5% benzylaminopurine and thiadiazole can effectively alleviate the inhibitory effect of nutritional stress on the growth of wheat seedlings.

[0181] 6.3 Effects of nutrient deficiency on wheat seedling height, root system, chlorophyll SPAD, and leaf area

[0182] Table 11 Effects of nutrient deficiency on plant height, root system, SPAD and leaf area of ​​wheat seedlings

[0183]

[0184]

[0185] Table 11 shows that application of a combined plant growth regulator of 0.5% benzylaminopurine and thiadipyridamole at FLC had little effect on wheat seedling height. However, application of this combined plant growth regulator significantly increased wheat plant height at FLU and TLU. Due to nutrient deficiencies, the root system gap between wheat seedlings with endosperm removal (CE and CER) and CK increased with increasing growth time. Wheat seedlings with both endosperm and root removal (CER) consistently lagged behind in root growth, but application of the agent (CER+Y) effectively alleviated this lag. Wheat seedlings with only root removal (CR) had significantly lower SPAD values ​​and total leaf area than seedlings with only endosperm removal (CE) at the time of one leaf extraction. However, as the plants grew, their SPAD values ​​and total leaf area exceeded those of CE at TLU. The SPAD values ​​and total leaf area of ​​the normally grown plants (CK) were the highest at all stages, while those of the wheat seedlings with both endosperm and root removed (CER) were the lowest.

[0186] 6.4 Effects of nutrient deficiency on dry matter accumulation in wheat seedlings

[0187] Table 12 Effects of nutrient deficiency on soluble protein and proline content in wheat seedlings

[0188]

[0189]

[0190] In Table 12, at the FLC and FLU stages, the dry weight of wheat seedlings with root removal (CR), endosperm removal (CE), and both root and endosperm removal (CER) was significantly lower than that of normally grown plants. As the plants grew, the root-removed wheat seedlings grew rapidly, while the endosperm removal (CE) and root and endosperm removal (CER) grew slowly, especially the wheat seedlings with both root and endosperm removal. After applying a 0.5% benzylaminopurine and thidiazuron combined plant growth regulator (CE+Y, CR+Y, CER+Y), their dry weights increased significantly compared to those not treated with the 0.5% benzylaminopurine and thidiazuron combined plant growth regulator (CE, CR, CER).

[0191] 6.5 Effects of nutrient deficiency on soluble protein and proline content in wheat seedlings

[0192] Figure 1 、 2The effects of nutrient deficiency on the soluble protein content and proline content of wheat seedlings are given respectively. Figure 1 、 2 As can be seen from the data, soluble protein content in normally growing plants gradually decreased from the FLC to the TLU period, while soluble protein content in plants with only endosperm removal (CE) and with both root and endosperm removal (CER) initially increased and then decreased with increasing growth period. Wheat seedlings without endosperm removal (CK and CR) had significantly higher soluble protein content than those with endosperm removal (CE and CER) across all three growth periods. Furthermore, wheat seedlings with only root removal (CR) had significantly higher soluble protein content than those with normally growing plants across all three growth periods. Treatments treated with a 0.5% combined plant growth regulator of benzylaminopurine and thiadiazole (CE+Y and CER+Y) showed significantly higher soluble protein content than those treated with CE and CER from the FLC to the TLU period. The proline content in normally grown wheat seedlings showed a gradually decreasing trend, while the proline content in wheat seedlings treated with only root removal (CR), endosperm removal (CE), and root and endosperm removal (CER) first increased and then decreased.

[0193] 6.6 Effects of nutrient deficiency on malondialdehyde content in wheat seedlings

[0194] Figure 3 The effects of nutrient deficiency on the malondialdehyde content in wheat seedlings are given. Figure 3 It can be seen that the malondialdehyde content in wheat seedlings with only the root system removed (CR) and the endosperm removed (CE, CER) showed a gradually decreasing trend with the increase of the growth period, and the malondialdehyde content in the treatments with 0.5% benzylaminopurine and thiadiazole-methyl composite plant growth regulator (CR+Y, CE+Y, CER+Y) increased.

[0195] 7 Conclusion

[0196] Wheat seedlings germinated for 48 hours were able to survive under the treatments of removing the endosperm, roots, and both the endosperm and roots. Wheat seedlings without endosperm grew slowly, with lower plant height, root length, SPAD, total leaf area, and dry weight, indicating that the endosperm plays an important role in maintaining nutrient supply. The soluble protein, proline, and malondialdehyde contents of wheat seedlings with the endosperm removed were low, which is not conducive to the normal growth of wheat. The soluble protein and proline contents of wheat seedlings with only the endosperm removed (CE), the endosperm and primary roots removed (CER), the endosperm removed and then the agent applied (CE+Y), and the endosperm and primary roots removed and then the agent applied (CER+Y) were significantly higher than those without the application of the combined plant growth regulator of 0.5% benzylaminopurine and thiadiazole. Soluble protein and proline provide nitrogen for wheat seedling growth, while accumulated malondialdehyde converts stored starch into glucose, which is beneficial for growth. This ultimately promotes rapid root and aerial growth, keeping it consistent with normal plant growth. In summary, applying a 0.5% compound plant growth regulator of benzylaminopurine and thiadiazole effectively alleviates the growth of wheat seedlings under nutritional stress and, to a certain extent, mitigates the growth inhibition caused by endosperm loss.

[0197] Application experiment 3: Effects of plant growth regulators on the growth of wheat seeds and seedlings under HgCl2 stress

[0198] 1. Experimental purpose: To clarify that plant growth regulators can effectively reduce the inhibitory effects of HgCl2 at different concentrations on wheat seed germination and seedling physiology, which has a very high guiding role in planting winter wheat in areas contaminated by heavy metal mercury.

[0199] 2. Experimental Design

[0200] 1. Test materials and reagents

[0201] 1.1 Test materials

[0202] Zhengmai 9023, Jimai 229, Hongdi 95 (winter wheat varieties)

[0203] 1.2 Test agent, same as above

[0204] 2 Test methods

[0205] 2.1 Chemical treatment

[0206] Weigh 0.1020 g of benzylaminopurine technical and 0.1020 g of thidiazuron technical, respectively, dissolve them in 1 ml of N-dimethylformamide (DMF), and dilute them with 0.1% Tween-80 aqueous solution to prepare mother liquors for later use; dilute the prepared benzylaminopurine mother liquor to 4.5 mg / L, and dilute the prepared thidiazuron mother liquor to 0.5 mg / L.

[0207] Preparation of a 0.5% benzylaminopurine and thidiazuron mixture: 100 ml of 4.5 mg / L benzylaminopurine and 0.5 mg / L thidiazuron were mixed and used as a stock solution. The solution was diluted with a 0.1% Tween-80 aqueous solution to a concentration of 0.5% (mass percentage) of the active ingredient before application. Treatments were indicated by Y, with a 0.1% Tween-80 aqueous solution serving as the CK control.

[0208] 2.2 Experimental design

[0209] Select pest-free, uniformly plump wheat seeds, disinfect them with 5% NaClO for 10 minutes, and rinse them five times with distilled water. Soak the seeds in a mixture of 0.5% benzylaminopurine and thiadiazole for 24 hours. A blank control (CK) is used for soaking seeds in a 0.1% Tween-80 aqueous solution. After soaking, rinse them three times with distilled water. Evenly distribute the seeds in a Petri dish lined with two layers of soaked filter paper and treat them as follows:

[0210] (1) Soak seeds with a mixture of 0.5% benzylaminopurine and thiadiazole, and soak filter paper with 50 mg / L, 100 mg / L, 150 mg / L, 200 mg / L, and 250 mg / L HgCl2, respectively;

[0211] (2) Soaking seeds in 0.1% Tween-80 aqueous solution, and soaking filter paper with 50 mg / L, 100 mg / L, 150 mg / L, 200 mg / L, and 250 mg / L HgCl2 respectively;

[0212] (3) Soak the seeds in 0.1% Tween-80 aqueous solution and soak the filter paper in 0.1% Tween-80 aqueous solution (CK).

[0213] The above treatments were repeated three times, with 100 seeds per group. To prevent excessive evaporation, the culture dishes were covered with plastic wrap. To maintain HgCl2 stability, the filter paper was replaced every two days. Seeds were cultured in an artificial intelligence climate chamber (RXZ-380) with a 12h / 12h light / dark cycle and 25±1°C. The number of seeds that germinated was recorded daily (using radicle length exceeding half the seed length as the standard). Germination potential was calculated on the third day, germination rate on the seventh day, and root length, shoot length, fresh weight, and physiological and biochemical parameters of the seedlings were measured on the eighth day.

[0214] 3 Measurement indicators

[0215] 3.1 Determination of seed germination and growth indicators

[0216] Germination potential = number of germinated seeds on the 3rd day / number of test seeds × 100%;

[0217] Germination rate = number of germinated seeds on the 7th day / number of test seeds × 100%;

[0218] Root length: the length from the root-bud junction to the longest root tip (root length was measured by a root scanner);

[0219] Bud length: the length from the base point to the bud tip;

[0220] Fresh weight: Use filter paper to absorb the surface moisture of the seedlings and weigh the fresh weight using an electronic balance.

[0221] 3.2 Determination of physiological and biochemical indicators

[0222] The peroxidase (POD) activity was determined by the guaiacol colorimetric method; the superoxide dismutase (SOD) activity was determined by the nitroblue tetrazolium method; the malondialdehyde (MDA) content was determined by the thiobarbituric acid (TBA) colorimetric method; and the soluble protein content was determined by Coomassie Brilliant Blue G-250 staining.

[0223] 4 Data Processing

[0224] DPS 9.01 was used for 5% significance comparison and Duncan's method was used for statistical analysis of the experimental data. Excel 2023 software was used for data collection, regression analysis and chart preparation.

[0225] 5 Test results

[0226] 5.1 Effects of plant growth regulator soaking on wheat seed germination under different HgCl2 concentrations

[0227] Figure 4 、 5 The effects of soaking seeds with or without pesticides on the germination rate of wheat seeds under different concentrations of HgCl2 stress are given respectively. Figure 4 and Figure 5As can be seen, in the blank control group, without the application of a 0.5% benzylaminopurine and thiadiazole combined plant growth regulator, the germination rates of Zhengmai 9023, Jimai 229, and Hongdi 95 were 85.33%, 87.67%, and 87.00%, respectively, in the absence of seed soaking with a 0.5% benzylaminopurine and thiadiazole combination plant growth regulator. After treatment with 150 mg / L HgCl₂, the germination rates of Zhengmai 9023, Jimai 229, and Hongdi 95 were significantly reduced to 79.20%, 78.37%, and 71.07%, respectively, indicating that 150 mg / L HgCl₂ significantly inhibited seed germination of the three wheat varieties. Under 150 mg / L HgCl₂ stress, the germination rates of Zhengmai 9023, Jimai 229, and Hongdi 95 decreased with increasing HgCl₂ concentration. The lowest germination rate was achieved at 250 mg / L HgCl₂, significantly lower than that of the blank control. The results showed that the effect of 250 mg / L HgCl2 stress on the germination rate of Zhengmai 9023, Jimai 229 and Hongdi 95 wheat seeds could not be alleviated without applying the combined plant growth regulator of 0.5% benzylaminopurine and thiadizuron for seed soaking. However, the germination rate of Zhengmai 9023, Jimai 229 and Hongdi 95 wheat seeds under HgCl2 stress could be significantly improved when the combined plant growth regulator of 0.5% benzylaminopurine and thiadizuron was applied for seed soaking. The germination rate of wheat seeds first increased and then decreased with the increase of HgCl2 concentration. The germination rate was the highest when the HgCl2 concentration was 100 mg / L, which was significantly different from the CK treatment group where the combined plant growth regulator of 0.5% benzylaminopurine and thiadizuron was not applied for seed soaking.

[0228] Figure 6 、 7 The effects of soaking seeds with or without pesticides on the germination potential of wheat seeds under different concentrations of HgCl2 stress are given respectively. Figure 6 、 7 As can be seen, in the absence of a 0.5% benzylaminopurine and thiadipyridinium combined plant growth regulator for seed soaking, germination potential of Zhengmai 9023, Jimai 229, and Hongdi 95 significantly decreased after 200 mg / L HgCl₂ treatment compared to the blank control, by 52.13%, 42.13%, and 33.97%, respectively. This indicates that 200 mg / L HgCl₂ stress significantly inhibits seed germination in these three wheat varieties. When treated with a 0.5% benzylaminopurine and thiadipyridinium combined plant growth regulator, germination potential and germination rate of Zhengmai 9023, Jimai 229, and Hongdi 95 initially increased and then decreased with increasing HgCl₂ concentration. In summary, seed soaking with a 0.5% benzylaminopurine and thiadipyridinium combined plant growth regulator effectively alleviates the toxic effects of HgCl₂ stress on wheat seed germination.

[0229] 5.2 Effects of plant growth regulator seed soaking on wheat seedling growth under different HgCl2 concentrations

[0230] Figure 8 、 9 The effects of seed soaking with and without pesticides on the root length of wheat seedlings under different concentrations of HgCl2 stress are given respectively. Figure 10 、 11 The effects of seed soaking with and without pesticides on the sprout length of wheat seedlings under different concentrations of HgCl2 stress are given respectively. Figure 12 、 13 The effects of seed soaking with and without pesticides on the fresh weight of wheat seedlings under different concentrations of HgCl2 stress are given respectively.

[0231] from Figure 8-13 It can be seen that compared with the blank control in the seed soaking treatment without the application of the drug, the root length, shoot length and fresh weight of the seedlings of Zhengmai 9023, Jimai 229 and Hongdi 95 were significantly reduced after treatment with 200 mg / L HgCl2. The root length, shoot length and fresh weight of Zhengmai 9023 were 3.11 cm, 3.44 cm and 0.09 g, respectively; those of Jimai 229 were 4.36 cm, 2.17 cm and 0.11 g, respectively; and those of Hongdi 95 were 3.44 cm, 4.34 cm and 0.11 g, respectively. This indicates that 200 mg / L HgCl2 stress can significantly inhibit the growth of the seedlings of the three wheat varieties.

[0232] Under different concentrations of HgCl2 stress, the root and shoot lengths of Zhengmai 9023, Jimai 229, and Hongdi 95 decreased significantly, while their fresh weights first increased and then decreased with increasing HgCl2 concentrations, indicating that 200 mg / L HgCl2 stress inhibited the growth of seedlings of Zhengmai 9023, Jimai 229, and Hongdi 95. After seed soaking with the agent, the root lengths of Zhengmai 9023, Jimai 229, and Hongdi 95 increased significantly with increasing HgCl2 concentrations, with the greatest increase at 150 mg / L. Shoot length and fresh weight did not increase significantly, and decreased significantly at some concentrations, indicating that seed soaking with the agent can promote wheat root growth under HgCl2 stress, but HgCl2 concentrations exceeding 150 mg / L cannot alleviate the toxic effects of HgCl2 stress on shoots and fresh weight.

[0233] 5.3 Effects of plant growth regulator seed soaking on physiological parameters of wheat seedlings under different HgCl2 concentrations

[0234] 5.3.1 Effects of Plant Growth Regulator Seed Soaking on SOD Content in Wheat Seedlings Under Different HgCl2 Concentrations

[0235] Figure 14 、 15 The effects of seed soaking with and without pesticides on the SOD content of wheat seedlings under different concentrations of HgCl2 stress are given respectively. Figure 14-15It can be seen that compared with the blank control in which no plant growth regulator was applied, the SOD activities of Zhengmai 9023, Jimai 229, and Hongdi 95 were significantly reduced after treatment with 150 mg / L HgCl2, reaching 114.31 U / g, 124.60 U / g, and 110.51 U / g, respectively. After soaking the seeds with plant growth regulators, the SOD activities of the three wheat varieties were significantly increased, reaching 124.33 U / g, 126.81 U / g, and 127.61 U / g, respectively.

[0236] 5.3.2 Effects of Seed Soaking with 0.5% Benzylaminopurine and Thidiazuron Combined Plant Growth Regulator on POD Content in Wheat Seedlings Under Different HgCl2 Concentrations

[0237] Figure 16 、 17 The effects of seed soaking with or without the application of pesticides on the POD content of wheat seedlings under different concentrations of HgCl2 stress are given respectively. Figure 16-17 Compared with the blank control (no plant growth regulator application), the POD activities of Zhengmai 9023, Jimai 229, and Hongdi 95 were 232.97 U / g, 221.05 U / g, and 202.04 U / g, respectively, after treatment with 150 mg / L HgCl2. The POD activities of all three wheat varieties increased significantly after seed soaking with plant growth regulators. The maximum POD activity of Zhengmai 9023 reached 261.13 U / g at 150 mg / L HgCl2, the maximum POD activity of Jimai 229 reached 312.01 U / g at 150 mg / L HgCl2, and the maximum POD activity of Hongdi 95 reached 241.06 U / g at 150 mg / L HgCl2. The results showed that seed soaking with plant growth regulators could significantly increase the SOD and POD activities of wheat seedlings under HgCl2 stress and enhance the antioxidant capacity of wheat seedlings.

[0238] 5.3.3 Effects of Plant Growth Regulator Seed Soaking on MDA Content in Wheat Seedlings Under Different HgCl2 Concentrations

[0239] Figure 18 、 19 The effects of seed soaking with or without the application of pesticides on the MDA of wheat seedlings under different concentrations of HgCl2 stress are given respectively. Figure 18-19Compared to the blank control (no plant growth regulator application), MDA content in Zhengmai 9023, Jimai 229, and Hongdi 95 significantly increased after treatment with 50-150 mg / L HgCl₂, indicating that HgCl₂ treatment can induce membrane lipid peroxidation in wheat seedlings. Following seed soaking with plant growth regulators, MDA content in Zhengmai 9023, Jimai 229, and Hongdi 95 initially increased and then decreased, reaching its lowest levels at 250 mg / L HgCl₂, at 0.00351 U / g, 0.00411 U / g, and 0.00331 U / g, respectively, demonstrating significant differences. This suggests that seed soaking with plant growth regulators can significantly reduce MDA content in wheat seedlings under HgCl₂ stress, alleviating membrane lipid peroxidation.

[0240] 5.3.4 Effects of Plant Growth Regulator Seed Soaking on Soluble Protein Content in Wheat Seedlings Under Different HgCl2 Concentrations

[0241] Figure 20 、 21 The effects of seed soaking with or without the application of pesticides on the soluble protein content of wheat seedlings under different concentrations of HgCl2 stress were given respectively. Figure 20-21 Compared to the control (no plant growth regulator application), the soluble protein contents of Zhengmai 9023, Jimai 229, and Hongdi 95 were 13.11 g, 13.11 g, and 13.31 g, respectively, after seed soaking with 200 mg / L HgCl₂. After seed soaking with plant growth regulators, the soluble protein content of all three wheat varieties initially increased and then decreased. The highest soluble protein contents were 25.70 g, 26.11 g, and 26.91 g, respectively, for Zhengmai 9023, Jimai 229, and Hongdi 95 at 200 mg / L HgCl₂. This suggests that seed soaking with plant growth regulators increased the soluble protein content of wheat seedlings under HgCl₂ stress.

[0242] 6. Summary

[0243] In this experiment, 50mg / L-100mg / L HgCl2 can promote the germination of wheat seeds to a certain extent, but under the stress of high concentration of 150mg / L-200mg / L HgCl2, it can cause greater damage to wheat seed germination and seedling growth. The application of plant growth regulators for seed soaking can alleviate this damage, improve the germination potential and germination rate of wheat, increase the soluble protein content, and induce the production of SOD and POD antioxidant enzymes in the seedlings, clear ROS in time, reduce the MDA content, alleviate oxidative damage, and thus reduce the damage of HgCl2 to wheat seedlings.

[0244] In summary, the plant growth regulator composition comprising benzylaminopurine and thidiazuron of the present invention can regulate wheat growth, effectively increasing the number of effective ears, the number of grains per ear, and the thousand-grain weight, improving agronomic traits, while also significantly increasing wheat yield and improving wheat quality. The plant growth regulator composition of the present invention can also be used to alleviate nutritional stress and heavy metal mercury stress in wheat.

Claims

1. A plant growth regulator composition for promoting wheat seed germination and seedling growth under nutritional stress, characterized in that: The plant growth regulator composition is composed of active ingredients benzylaminopurine and thidiazuron, and the mass ratio of the benzylaminopurine to thidiazuron is (40-50):

5.

2. The use according to claim 1, characterized in that The mass ratio of benzylaminopurine to thiadiazole is (43-47):

5.

3. The use according to claim 1, characterized in that The plant growth regulator composition is made of active ingredients and pesticide adjuvants into a dosage form allowed by pesticides, and the dosage form is wettable powder, soluble powder, soluble liquid, water-dispersible granules, suspension, emulsifiable concentrate, water emulsion, microemulsion or microcapsule.

4. The use according to any one of claims 1 to 3, characterized in that The total content of the active ingredients accounts for 0.3-0.7% of the total weight of the entire plant growth regulator composition.

5. The use according to claim 1, characterized in that The plant growth regulator composition is diluted with water and then used to soak wheat seeds, thereby increasing the growth rate, plant height, root length, SPAD, total leaf area and dry weight of the wheat.

6. Use of a plant growth regulator composition in promoting the growth of wheat seeds and seedlings under HgCl2 stress, characterized in that: The plant growth regulator composition is composed of active ingredients benzylaminopurine and thidiazuron, and the mass ratio of the benzylaminopurine to thidiazuron is (40-50):

5.

7. The use according to claim 6, characterized in that The mass ratio of benzylaminopurine to thiadiazole is (43-47):

5.

8. The use according to claim 6, characterized in that The plant growth regulator composition is made of active ingredients and pesticide adjuvants into a dosage form allowed by pesticides, and the dosage form is wettable powder, soluble powder, soluble liquid, water-dispersible granules, suspension, emulsifiable concentrate, water emulsion, microemulsion or microcapsule.

9. The use according to any one of claims 6 to 8, characterized in that The total content of the active ingredients accounts for 0.3-0.7% of the total weight of the entire plant growth regulator composition.

10. The use according to claim 6, characterized in that The plant growth regulator composition is diluted with water and then used to soak wheat seeds, thereby improving the germination potential, germination rate and soluble protein content of the wheat.

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