Application of thiabenuron in improving heat tolerance of anthurium andraeanum

Abstract

The application provides application of thidiazuron in improving heat resistance of anthurium, and relates to the technical field of plant growth regulators.The application applies thidiazuron to improve heat resistance of anthurium, can effectively delay the discoloring process of the spathes of anthurium under high-temperature stress, maintains good ornamental properties of the anthurium, and improves the resistance of the anthurium to high-temperature stress by improving antioxidant enzyme activity and protecting cell membrane integrity, so that the anthurium grows well.The application can effectively improve the heat resistance of the anthurium by applying exogenous thidiazuron, the application method is simple, the cost is low, and the application is suitable for the production of anthurium in high-temperature areas such as Hainan in summer.

Description

Technical Field

[0001] This invention relates to the field of plant growth regulator technology, and in particular to the application of thiabendazole in improving the heat resistance of Anthurium. Background Technology

[0002] Anthurium ( Anthurium andraeanum ) belongs to the genus Anthurium in the family Araceae. Anthurium Anthurium is a perennial herbaceous plant with beautiful flowers and leaves, unique flower shape, and high ornamental value. Anthurium prefers a warm, humid, and semi-shaded environment. The suitable growth temperature is 18~30℃, the overwintering temperature should not be lower than 15℃, and the summer temperature should not be higher than 32℃, otherwise it will cause poor growth and fading of flowers and leaves.

[0003] In recent years, Hainan has experienced persistently high summer temperatures, which has affected the growth and development of Anthuriums, resulting in phenomena such as discoloration of the spathe and shortened flower lifespan. Current research focuses primarily on the cultivation, propagation, and breeding of Anthuriums, with relatively little research on improving their heat tolerance. Summary of the Invention

[0004] Therefore, the purpose of this invention is to propose the application of thiamethoxam in improving the heat resistance of Anthurium, which can effectively improve the growth status of Anthurium under high temperature stress, solve problems such as fading of Anthurium bracts, cell membrane damage, and decreased antioxidant enzyme activity under high temperature, and is simple to operate and has stable effects.

[0005] The technical solution of this invention is implemented as follows: This invention provides the application of thiazuron (TDZ) in improving the heat resistance of Anthurium, wherein the Anthurium is *Pink Crown*. The thiamethoxam of the present invention improves the heat resistance of Anthurium andraeanum, specifically achieving the following effects: (1) Thiabenone delays the fading process of Anthurium under high temperature stress and effectively prolongs the ornamental life of Anthurium flowers; (2) Thiidianone increases the antioxidant enzyme activity of Anthurium under high temperature stress, enhances Anthurium's own reactive oxygen scavenging ability, and reduces oxidative damage; (3) Thiidianone protects the cell membrane integrity of Anthurium under high temperature stress, reduces the leakage of intracellular solutes, and maintains normal cell physiological function.

[0006] Furthermore, the antioxidant enzyme is superoxide dismutase and / or peroxidase; antioxidant enzymes are the core components for plants to scavenge reactive oxygen species. Exogenous application of thiamethoxam can enhance the activity of the enzymes mentioned above in Anthurium, thereby enhancing the antioxidant capacity of Anthurium and improving its heat resistance.

[0007] Furthermore, the concentration of thiamethoxam used is 1~1.5 mg / L.

[0008] Furthermore, the method of using thiamethoxam to improve the heat resistance of Anthurium is as follows: before high temperature stress, apply thiamethoxam solution to the bracts and leaves of Anthurium and perform root irrigation with thiamethoxam solution to improve the resistance of Anthurium to high temperature stress. Different application methods can all achieve a good effect on improving the heat resistance of Anthurium.

[0009] Furthermore, spray the bracts and leaves of Anthurium with thiamethoxam solution for 5-10 consecutive days; And every 1 to 2 days, drench the roots with thiamethoxam solution once; The above application frequency ensures that Anthurium fully absorbs thiabendazole, achieving a stable regulatory effect.

[0010] Furthermore, the dosage for root drenching is 150-250 mL, the treatment cycle is 5-10 days, and the total number of root drenching treatments is 3-4 times.

[0011] The present invention also provides a method for improving the heat resistance of Anthurium using thiazuron. Before high temperature stress, the bracts and leaves of Anthurium are sprayed with a 1-1.5 mg / L thiazuron solution for 5-10 consecutive days. Every 1-2 days, the roots were drenched with a 1-1.5 mg / L thiamethoxam solution for 5-10 days. The red palm is actually a pink crown.

[0012] Furthermore, the dosage for root drenching is 150-250 mL, and the total number of root drenching treatments is 3-4.

[0013] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention applies thiamethoxam to improve the heat resistance of Anthurium, which can effectively delay the fading process of the spathe of Anthurium under high temperature stress, maintain the good ornamental characteristics of Anthurium, and at the same time improve the resistance of Anthurium to high temperature stress and promote good growth and development by enhancing antioxidant enzyme activity and protecting cell membrane integrity.

[0014] This invention provides a method to effectively improve the heat resistance of Anthurium by applying thiamethoxam exogenously. The application method is simple and low-cost, making it suitable for Anthurium production in high-temperature summer regions such as Hainan. Attached Figure Description

[0015] Figure 1 Phenotypic changes of *Cephalotaxus fortunei* bracts treated with different concentrations of thiamethoxam at high temperatures; Figure 2 The graph shows the chroma changes of pink bracts treated with different concentrations of thiamethoxam at high temperature. * indicates a significant difference (Tukey test, *, p <0.05; **,p <0.01,***, p <0.001), compared with the control group, the data are mean ± SD, n=4; Figure 3 The graph shows the electrical conductivity of *Pterocarya stenoptera* leaves after 120 h of high-temperature treatment with different concentrations of thiamethoxam. Different lowercase letters represent Tukey's test results. p <0.05) indicates a significant difference; the same letter indicates no significant difference. Figure 4 The graph shows the changes in superoxide dismutase activity in *Cephalotaxus fortunei* bracts after 120 h of high-temperature treatment with different concentrations of thiabendazole. Different lowercase letters indicate Tukey's test results. p <0.05) indicates a significant difference; the same letter indicates no significant difference. Figure 5 The graph shows the changes in peroxidase activity in *Cephalotaxus fortunei* bracts after 120 h of high-temperature treatment with different concentrations of thiabendazole. Different lowercase letters indicate Tukey's test results. p <0.05) indicates a significant difference; the same letter indicates no significant difference. Detailed Implementation

[0016] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0017] Unless otherwise specified, the experimental methods used in the embodiments of this invention are all conventional methods.

[0018] Unless otherwise specified, all materials and reagents used in the embodiments of this invention are commercially available.

[0019] Color saturation c Method for determining the value: Select 3 spathes from each pot, and measure the value at the middle of the left and right sides of the spathes respectively. a Value and b The value was measured three times. a Value and b The values ​​were measured using a colorimeter. a Value and b The value represents two dimensions of color, with each value corresponding to a color attribute. a Value (Red-Green Bias): Specifically distinguishes between red and green tones, ranging from -128 to +128. Positive values ​​indicate a red bias, while negative values ​​indicate a green bias. b Value (Yellow / Blue Bias): Defines the yellow and blue hues, ranging from -128 to +128. Positive values ​​lean towards yellow, while negative values ​​lean towards blue. Color Scalec The value is based on the formula for color saturation. Perform the calculation.

[0020] Methionine (Met), Nitrotetrazolium Bluechloride (NBT), and Ethylenediaminetetraacetic acid disodium salt (EDTA-Na2).

[0021] Thiabenone was purchased from Haikou Longhua Chuangyuan Biotechnology Center.

[0022] Peroxidase (POD) activity assay kit 200T / box and superoxide dismutase (SOD) activity assay kit 200T / box were purchased from Haikou Jirui Biotechnology Co., Ltd.

[0023] Pink Crown comes from the Anthurium resource nursery of the Institute of Tropical Crop Germplasm Resources, Chinese Academy of Tropical Agricultural Sciences.

[0024] Example 1 1. Materials and Methods 1.1 Test Materials Choose the Pink Crown variety, which is prone to fading at high temperatures.

[0025] 1.2 Test Methods Treatments were administered with 0.5 mg / L, 1.0 mg / L, and 1.5 mg / L TDZ (thiabendazole, Solarbio, T8051-1g) for 7 consecutive days. Before the high-temperature treatment, the bracts and leaves were sprayed until moist. During these 7 days, the roots were irrigated once every other day, with 200 mL per pot each time.

[0026] Set up a high-temperature control group (CK): treat with distilled water for 7 consecutive days. Spray the bracts and leaves until moist before the high-temperature treatment. Water the roots once every day during these 7 days, 200 mL per pot each time.

[0027] Preparation of 1 mg / mL TDZ stock solution: Weigh 10.0 mg of TDZ powder; add 0.5~1 mL of 0.1 mol / L NaOH, gently vortex until completely dissolved; bring the volume to 10 mL with distilled water; filter through a 0.22 μm filter membrane for sterilization. Preparation of 0.5 mg / L, 1.0 mg / L, and 1.5 mg / L TDZ solutions: Take 0.5 mL, 1.0 mL, and 1.5 mL of the 1 mg / mL TDZ stock solution, respectively, add 1 L of distilled water, and mix well.

[0028] The high-temperature treatment conditions were high temperature (photocycle, 12 h / 12 ​​h; day / night temperature of 38℃ / 30℃; light intensity of 12000 lx / 0 lx; ​​humidity of 75%), and the soil was kept moist during the cultivation period.

[0029] A control group (N) was set up at room temperature: the room temperature treatment conditions were room temperature (photocycle, 12 h / 12 ​​h; day and night temperature, 25℃ / 25℃; light intensity, 12000 lx / 0 lx; ​​humidity, 75%), and the soil was kept moist during the cultivation period.

[0030] The color intensity of the anthurium bracts was measured after 0 h, 48 h, and 120 h of experimental treatment. And sampling was performed, with each time point repeated 3 times.

[0031] The relative electrical conductivity of Anthurium leaves and the antioxidant enzyme activity of Anthurium bracts were measured 120 h after the experimental treatment. Each time point was repeated 3 times.

[0032] 1.2.1 Relative conductivity measurement After high-temperature treatment, several small circular pieces were taken from the leaf using a perforator, and 0.1 g was accurately weighed and placed in a 50 mL centrifuge tube. 25 mL of ultrapure water was added. After shaking at 80 r / min for 1 h at room temperature, the initial conductivity was measured using a conductivity meter and recorded as E1. The sample was then placed in a 95 ℃ water bath for 30 min, cooled to room temperature, and the conductivity was measured again and recorded as E2. The relative conductivity (REC) was calculated using the following formula: REC = E1 / E2 × 100%.

[0033] 1.2.2 Antioxidase activity assay Superoxide dismutase (SOD) assay (1) Extraction of enzyme solution: Weigh 0.05 g of bracts, add 900 μL of pre-cooled 0.05 mol / L phosphate buffer (pH 7.0), grind on ice, extract for 2 h, centrifuge at 10000 g for 15 min at 4 ℃, and the supernatant is the enzyme solution. Transfer to a new centrifuge tube and place on ice.

[0034] (2) Add the various solutions according to the table below, with riboflavin added last. The working solution can be prepared according to the number of samples. Add 200 μL of working solution to each well of the microplate. Set up a blank control by replacing the enzyme solution with an equal volume of 0.05 mol / L phosphate buffer (pH 7.0).

[0035] Table 1. Solution preparation for superoxide dismutase determination

[0036] Each treated sample was placed under approximately 4000 lx light for 25 min. The specific reaction time could be optimized based on preliminary experimental results. After the reaction, the absorbance of each sample was measured at 560 nm using a microplate reader, and the superoxide dismutase (SOD) activity was calculated accordingly.

[0037] The calculation formula is as follows: SOD (U / g) = [( Ack-AE )×V ] / ( 1 / 2Ack×m×Vt ) Where: Ack—absorbance of the control tube AE—Absorbance of the sample tube V — Total volume of sample solution (mL) m — Fresh weight of sample (g) Vt — Volume of enzyme solution used in the assay (mL) Peroxidase (POD) activity was measured using the Solarbio POD Activity Assay Kit. Each sample group was tested in triplicate. Experimental procedures and enzyme activity calculations were performed according to the kit instructions.

[0038] 1.3 Results and Analysis 1.3.1 Phenotypic Changes of Anthurium Bracts under High Temperature Treatment with Exogenous Substances Depend on Figure 1 It was found that under high-temperature stress, the bracts of the high-temperature control group (CK) continued to fade with prolonged treatment time. Treatment with thiamethoxam at various concentrations could delay bract fading, with 1.5 mg / L showing the best effect in maintaining the red color of the bracts, while 0.5 mg / L had the weakest mitigating effect, indicating that the appropriate concentration of thiamethoxam can effectively inhibit bract fading caused by high temperature.

[0039] Depend on Figure 2It was found that under high-temperature stress, the bracts of Anthurium exhibited significant fading, but TDZ treatment significantly delayed this process. The fading rate was calculated based on the 0-hour color intensity of each treatment. The results showed that the high-temperature control group (CK) bracts showed the most significant fading at 48 h and 120 h, with fading rates of 18.0% and 28.3%, respectively; the 0.5 mg / L TDZ group showed fading rates of 14.7% and 23.7%, respectively, only partially delaying fading; the 1.0 mg / L TDZ group showed fading rates of 6.2% and 14.8%, respectively, demonstrating a significant color-preserving effect; the 1.5 mg / L TDZ group showed the smallest fading rate (4.4% at 48 h and 10.7% at 120 h), with the most vibrant and uniform red bract color. Phenotypic observations were consistent with the color difference data, indicating that the color-preserving effect of TDZ treatment was concentration-dependent, with 1.5 mg / L TDZ showing the best effect in delaying high-temperature-induced bract fading.

[0040] 1.3.2 Changes in electrical conductivity of Anthurium leaves under high-temperature treatment with exogenous substances Depend on Figure 3 The results showed that the average REC of the room temperature control group (N) was 12.49%, indicating good leaf membrane integrity and no high-temperature damage. The REC of the high-temperature control group (CK) was significantly increased to 16.34%, indicating that high-temperature stress damaged the cell membrane structure and caused significant membrane damage. The REC of 0.5 mg / L TDZ was 13.16%, lower than that of the high-temperature control group (CK), indicating that low-concentration TDZ had a partial protective effect on the membrane. The REC of 1.0 mg / L TDZ was the lowest (12.52%), close to the room temperature level, indicating that medium-concentration TDZ provided the best membrane protection and the least high-temperature damage. The REC of 1.5 mg / L TDZ was 12.74%, still lower than that of the high-temperature control group (CK), but higher than that of 1.0 mg / L, indicating that the protective effect of high-concentration TDZ on the membrane was slightly weakened.

[0041] 1.3.3 Changes in superoxide dismutase activity in anthurium bracts treated with exogenous substances at high temperatures Depend on Figure 4It was found that high-temperature stress could induce an antioxidant response in the bracts of the high-temperature control group (CK), resulting in enhanced SOD activity. Higher SOD activity indicates stronger antioxidant capacity. The average SOD activity of the high-temperature control group (CK) was 338.8 U / g, which was about 17.8% higher than that at room temperature, indicating that high-temperature stress induced an antioxidant response in the bracts, but it may still be insufficient to completely resist oxidative stress. The average SOD activity of 0.5 mg / L TDZ was 310.1 U / g, which was slightly lower than that of the high-temperature control group (CK) but slightly higher than that at room temperature, indicating that low concentrations of TDZ had limited effect on increasing SOD activity and had only moderate antioxidant capacity. The average SOD activity of 1.0 mg / L TDZ was 416.6 U / g, which was significantly higher than that of both the high-temperature control group (CK) and the room-temperature control group (N), indicating that high concentrations of TDZ significantly enhanced the antioxidant capacity of the bracts and effectively scavenged superoxide anions. The average SOD activity of 1.5 mg / L TDZ was 436.7 U / g, the highest among all treatments, indicating that high concentrations of TDZ had the strongest stimulating effect on SOD activity and the strongest antioxidant capacity of the bracts.

[0042] 1.3.4 Changes in peroxidase activity in Anthurium bracts after treatment with exogenous substances at high temperature Depend on Figure 5 The results showed that the average POD activity of the high-temperature control group (CK) was 49.00 U / g, a decrease of approximately 22.6% compared to the normal temperature control group, indicating that high-temperature stress reduced the bracts' ability to scavenge H2O2 and increased oxidative stress. The average POD activity of 0.5 mg / L TDZ was 29.47 U / g, lower than both the high-temperature control group (CK) and the normal temperature control group (N), indicating that low-concentration TDZ did not adequately stimulate POD activity and reduced antioxidant capacity. The average POD activity of 1.0 mg / L TDZ was 60.22 U / g, higher than the high-temperature control group (CK) but slightly lower than the normal temperature control group (N), showing that medium-concentration TDZ could enhance the bracts' ability to scavenge H2O2 while maintaining the natural color of the bracts. The average POD activity of 1.5 mg / L TDZ was 72.67 U / g, the highest among all treatments, indicating that high-concentration TDZ had the strongest stimulating effect on POD activity and also enhanced the antioxidant capacity of the bracts.

[0043] In summary, under high-temperature stress, the bracts of Anthurium exhibited significant fading, but TDZ treatment significantly delayed this process. The fading rate was calculated based on the 0-hour color intensity of each treatment. The results showed that the high-temperature control group (CK) bracts showed the most significant fading at 48 h and 120 h, at 18.0% and 28.3% respectively; the 0.5 mg / L TDZ group showed 14.7% and 23.7% respectively; the 1.0 mg / L TDZ group showed 6.2% and 14.8% respectively, with significantly reduced fading and a more natural and uniform color; the 1.5 mg / L TDZ group showed the smallest fading rate (4.4% at 48 h and 10.7% at 120 h), with the most vibrant bract color, slightly darker red.

[0044] Relative conductivity (REC) analysis showed that high temperature stress significantly increased leaf membrane permeability, with an average REC of 16.34% in the high temperature control group (CK) and 12.49% in the normal temperature control group (N). TDZ treatment showed a concentration-dependent effect on membrane stability: 0.5 mg / L TDZ had an average REC of 13.16%, partially protecting the membrane structure; 1.0 mg / L TDZ had the lowest average REC (12.52%), with membrane integrity close to that at room temperature, showing the best protective effect; 1.5 mg / L TDZ had an average REC of 12.74%, which, although still lower than the high temperature control group (CK), slightly reduced the membrane protection effect.

[0045] Further analysis of antioxidant enzyme activities showed that TDZ regulated the activities of SOD and POD in bracts in a concentration-dependent manner: SOD activity: 1.0 mg / L and 1.5 mg / L TDZ were 416.6 and 436.7 U / g, respectively, higher than the high-temperature control group (CK) (338.8 U / g), significantly enhancing the ability to scavenge superoxide anions; 0.5 mg / L TDZ activity was 310.1 U / g, with limited improvement. POD activity: 1.0 mg / L TDZ had an average of 60.22 U / g, higher than the high-temperature control group (CK) (49.00 U / g), improving antioxidant capacity; 1.5 mg / L TDZ had the highest activity (72.67 U / g), the strongest antioxidant capacity, and the bracts were slightly darker red; 0.5 mg / L TDZ activity was 29.47 U / g, with insufficient antioxidant capacity.

[0046] Example 2 Treat the powdery crown with 1.5 mg / L TDZ for 10 days. Before the high-temperature treatment, drench the roots once every day, 200 mL per pot each time.

[0047] The high-temperature treatment conditions were: high temperature (photocycle, 12 h / 12 ​​h; day / night temperature, 38℃ / 30℃; light intensity, 12000 lx / 0 lx; ​​humidity, 75%), with the soil kept moist during the cultivation period. Color intensity was measured after 0 h and 120 h of high-temperature treatment, with each time point repeated three times.

[0048] Example 3 Treat *Ceropegia woodii* with 1.5 mg / L TDZ for 7 consecutive days. Spray the bracts until moist before high-temperature treatment. During these 7 days, irrigate the roots once every other day, 200 mL per pot each time.

[0049] The high-temperature treatment conditions were: high temperature (photocycle, 12 h / 12 ​​h; day / night temperature, 38℃ / 30℃; light intensity, 12000 lx / 0 lx; ​​humidity, 75%), with the soil kept moist during the cultivation period. Color intensity was measured after 0 h and 120 h of high-temperature treatment, with each time point repeated three times.

[0050] Under high-temperature stress, the bracts of Anthurium in Examples 2 and 3 showed obvious fading. The fading range was calculated based on the 0-hour color intensity of each of Examples 2 and 3. After 120 hours of high-temperature treatment, the fading range was relatively small, and the bracts remained bright and uniformly red.

[0051] The above description represents the preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of the present invention, and these improvements and modifications are also considered to be within the scope of protection of the present invention.

Claims

1. The application of thiamethoxam in Anthurium cultivation, wherein the thiamethoxam is used to improve the heat resistance of Anthurium, wherein the Anthurium is *Pink Crown*. Thiidianone improves the heat resistance of Anthurium by including at least one of the following: (1) Thiabenzodone delays the fading process of Anthurium under high temperature stress; (2) Thiidianone increases the antioxidant enzyme activity of Anthurium under high temperature stress; (3) Thiabenone protects the cell membrane integrity of Anthurium under high temperature stress.

2. The application according to claim 1, characterized in that, The antioxidant enzyme is superoxide dismutase and / or peroxidase.

3. The application according to claim 1, characterized in that, The concentration of thiamethoxam used is 1~1.5 mg / L.

4. The application according to claim 1, characterized in that, The method to improve the heat resistance of Anthurium is as follows: before high temperature stress, apply thiazuron solution to the bracts and leaves of Anthurium, and then drench the roots with thiazuron solution to improve the resistance of Anthurium to high temperature stress.

5. The application according to claim 4, characterized in that, Spray the bracts and leaves of Anthurium with thiamethoxam solution for 5-10 consecutive days. The roots were drenched with thiamethoxam solution every 1-2 days.

6. The application according to claim 5, characterized in that, The dosage for root drenching is 150-250 mL, the treatment cycle is 5-10 days, and the total number of root drenching treatments is 3-4.

7. A method for improving the heat resistance of Anthurium using thiamethoxam, characterized in that, Before high temperature stress, spray the bracts and leaves of Anthurium with a 1-1.5 mg / L thiamethoxam solution for 5-10 consecutive days. Every 1-2 days, drench the roots with a 1-1.5 mg / L thiamethoxam solution once, for a period of 5-10 days. The red palm is actually a pink crown.

8. A method for improving the heat resistance of Anthurium using thiamethoxam according to claim 7, characterized in that, The dosage for root drenching is 150-250 mL, and the total number of root drenching treatments is 3-4.

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