Application and preparation of beta-sitosterol

A technology of sitosterol and pharmaceutical preparations, which is applied in the field of application of β-sitosterol and its preparations, and can solve problems such as unresearched

Active Publication Date: 2017-09-01

SICHUAN AGRI UNIV

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Abstract

The invention relates to the technical field of agricultural production and particularly relates to application and a preparation of beta-sitosterol, in particular to application of beta-sitosterol to improvement in plant drought resistance. An experiment shows that the beta-sitosterol can effectively delay the wilting of white clover leaves under drought stress, relieve the growth inhibition caused by drought, reduce the degradation, the per-oxidation damage and the like of chlorophylls and significantly improve the drought resistance of white clovers. An application effect is not limited by the application time, and the drought-resistant effects on drought sensitive white clover varieties and drought-resistant white clover varieties are better. In addition, compared with ABA, SA, brassinolide and other exogenous additives, the beta-sitosterol is cheap in price, low in production cost and suitable for large-area popularization and application.

Application Domain

BiocidePlant growth regulators +1

Technology Topic

Drought resistanceAgricultural science +8

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Examples

Experimental program(5)

Example Embodiment

[0017] Example 1 Screening of β-sitosterol root application concentration

[0018] Test material: The widely used 'Latino' white clover is used as the test material

[0019] Material cultivation: First, soak the seeds of white clover with 0.1% potassium permanganate solution for 10 minutes and disinfect them, then wash them with deionized water for 3 times. 2 The seeds were sown in plastic pots with a length of 20cm, a width of 15cm and a height of 5cm and were filled with quartz sand, and placed in a light incubator for germination (temperature day/night was 23/19°C; duration was 12h; relative Humidity is 70%; light intensity is 700 μmol m -2 s -1 ). After 7 days of seed germination, the seedlings were further cultivated with Hoagland nutrient solution for 30 days, and the materials with the same growth vigor were selected for subsequent experiments.

[0020] Experimental design: A total of 7 β-sitosterol concentration gradients were set, respectively: 0mg/L (control, direct drought stress), 5mg/L, 25mg/L, 50mg/L, 100mg/L, 250mg/L, 500mg/L L, 7 treatments in total, with 4 independent replicates for each treatment. Drought stress treatment was performed with 18% polyethylene glycol 6000 (PEG-6000).

[0021] Exogenous addition and drought stress treatment: β-sitosterol was dissolved in 100% hot ethanol (0.2 g/mL), cooled to room temperature naturally and used as a mother solution for later use. The nutrient solution was used as a solvent, and the β-sitosterol treatment solutions of different concentrations were prepared with the β-sitosterol mother solution. Hoagland full nutrient solution for drought stress treatment. During the treatment, the treatment solution was replaced every 3 days, 300 mL each time. All materials were placed in a smart light incubator under the same conditions as the materials. The timing was started when the treatment solution containing 18% (W/V) PEG-6000 was replaced, and the time of wilting symptoms and severe wilting of white clover leaves was investigated. The results are shown in Table.

[0022] Table 1 The effect of root application of different concentrations of β-sitosterol on improving the drought resistance of white clover

[0023]

[0024] It can be seen from Table 1 that the root application of 25-100 mg/L β-sitosterol can significantly improve the drought resistance of white clover, and the effect of improving the cold resistance of white clover is better. The wilting of white clover leaves can be delayed for 3-5 days under drought stress intensity.

Example Embodiment

[0025] Example 2 Screening of β-sitosterol concentration by foliar spraying

[0026] Test material: The drought-sensitive 'Latino' white clover was used as the test material.

[0027] Material cultivation: Mix the garden soil and the nutrient soil in a ratio of 1:1, add pesticides and fungicides to mix evenly, and store them outdoors for 3 days for later use. Use a pot with a diameter of 20cm and a depth of 25cm to plant white clover, fill the pot with the same amount of mixed soil, and press 1g/m 2 Sow the white clover seeds at the seeding rate, cover with shallow soil and water. The materials were placed in a greenhouse for 3 months, and 250 mL of Hoagland's total nutrient solution was poured into each pot every week. The average temperature of the greenhouse is 23℃, the relative humidity is 65%, the light time is 12h, and the light intensity is 500μmol m -2 s -1. After 7 days of seed germination, the seedlings were further cultivated with Hoagland nutrient solution for 30 days, and the materials with the same growth vigor were selected for subsequent experiments.

[0028] Experimental design: A total of 7 β-sitosterol concentration gradients were set, respectively: 0mg/L (control, direct drought stress), 5mg/L, 10mg/L, 15mg/L, 25mg/L, 50mg/L, 100mg/L L, 250 mg/L, a total of 8 treatments, each treatment set 4 independent repetitions.

[0029]Exogenous spraying and drought stress treatment: β-sitosterol was fully dissolved in 100% hot ethanol (0.2g/mL) as a mother liquor for standby use, and 1% Tween-80 was used as a solvent to dilute the mother liquor to prepare β-sitosterol solutions of different concentrations , the control was sprayed with the corresponding solvent without β-sitosterol. Foliar spraying was carried out at 9:00 am each time, the spraying amount was 150mL, and the spraying was carried out 3 times in a row, with an interval of 1 day each time. During the spraying of exogenous additives, normal watering was performed. After the treatment, all materials were stopped from watering and placed in a greenhouse for natural drought stress. Timing was started from natural drought stress (i.e., watering was stopped), and the time when wilting symptoms and severe wilting of white clover leaves appeared was investigated. The results are shown in Table 2.

[0030] Table 2 Effects of foliar spraying with different concentrations of β-sitosterol on improving drought resistance of white clover

[0031]

[0032] It can be seen from Table 2 that when 10-25mg/L of β-sitosterol was sprayed on the foliar surface, the drought resistance of white clover could be significantly improved. Delay 5-7 days.

Example Embodiment

[0033] Example 3 Effects of foliar spraying of β-sitosterol on physiological indicators of drought resistance in white clover

[0034] Test material: Drought-sensitive 'Latino' white clover was used as the test material

[0035] Material cultivation: same as Example 2

[0036] Experimental design: The optimal leaf spraying concentration screened out according to Experimental Example 2 is 15 mg/L as the experimental spraying concentration. A total of 4 treatments were set up in the experiment, and each treatment was set up with 4 independent repetitions, respectively: ① Normal condition control (CK ): After foliar spraying with a solvent without β-sitosterol, cultured in the greenhouse for 18 days as a control; ②CK+β-sitosterol: After foliar spraying with 15 mg/L β-sitosterol, normal culture in the greenhouse 18 days; ③Direct drought stress: After foliar spraying with a solvent without β-sitosterol, natural drought in the greenhouse for 18 days; ④Dry + β-sitosterol: After foliar spraying 15 mg/L β-sitosterol Naturally dry in a greenhouse for 18 days.

[0037] Drought stress treatment: foliar spraying was carried out at 9:00 in the morning for each spraying, and 100mL of the same amount of liquid was evenly sprayed on the foliar surface of each treatment until all the leaves were covered with water droplets, and sprayed three times continuously, each time 1 day interval. After the treatment, the control group (CK and CK+β-sitosterol) was watered normally, and the drought stress group (drought and drought+β-sitosterol) was stopped watering and placed in the greenhouse for natural drought stress. Greenhouse conditions were the same as in Example 2.

[0038] Observation indicators: When the drought stress was 18 days, the dry weight and relative growth of a single plant reflecting the growth status of white clover were measured; the relative water content reflecting the water status of leaves; Content, superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), peroxidase (POD) activities. The results are shown in Table 3.

[0039] Table 3 Changes of physiological and biochemical indexes related to drought resistance of white clover improved by foliar spraying of β-sitosterol (15mg/L)

[0040]

[0041] It can be seen from Table 3 that under normal conditions, spraying exogenous β-sitosterol pretreatment can promote the growth and chlorophyll synthesis of white clover plants; under drought stress, spraying exogenous β-sitosterol pretreatment can significantly increase the Alleviates the growth inhibition caused by drought, reduces the degradation of chlorophyll, and alleviates the peroxidative damage caused by drought by increasing the activities of various antioxidant enzymes, and significantly reduces the electrolyte permeability and the accumulation of the membrane lipid peroxidation product malondialdehyde. These growth and physiological and biochemical indicators further proved that foliar spraying of β-sitosterol pretreatment can effectively improve the drought resistance of white clover.

PUM

Property	Measurement	Unit
Diameter	20.0	cm
Depth	25.0	cm

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