Application of glycoside alkaloid compounds in prevention and / or treatment of plant bacterial leaf blight
An antibacterial agent prepared by using glycoside alkaloids extracted from the green fruit of *Solanum nigrum* has solved the problem of controlling bacterial leaf blight in rice, effectively inhibiting the disease and promoting rice growth, thus enhancing the disease resistance of rice.
Patent Information
- Application Number
- CN202610341047.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-03-19
- Publication Date
- 2026-07-03
Smart Images
Figure CN122320044A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the application of glycoside alkaloid compounds in the prevention and / or treatment of plant bacterial blight, and belongs to the field of agricultural technology. Background Technology
[0002] Bacterial blight (BB) in rice is caused by Xanthomonas aeruginosa. Xanthomonas oryzae pv. oryzae Bacterial blight (Bacterial blight) caused by Xylaria (Xoo) is one of the major diseases affecting rice cultivation in my country. This disease primarily damages the vascular bundles of rice, causing water-soaked lesions on the leaf tips and edges, which gradually expand into yellowish-brown necrotic streaks. In severe cases, it can cause leaf curling, wilting, and even the death of the entire plant. Bacterial blight is extremely harmful to rice, leading to yield reductions of 20% to 50%, and even exceeding 60% in epidemic years, seriously threatening global rice production and food security. With climate change and altered farming practices, the frequency and severity of this disease are continuously increasing.
[0003] Plant-derived active ingredients play a crucial role as antifungal agents in the control of agricultural diseases. These ingredients, derived from natural plants, offer advantages over chemically synthesized pesticides, including environmental friendliness, low residue, low toxicity to non-target organisms, high safety profile, and low likelihood of resistance development, thus meeting the demands of modern ecological agriculture for sustainable development. However, currently, there is a lack of plant-derived antifungal agents for controlling rice bacterial blight, necessitating the development of novel and highly effective plant-derived antifungal agents to effectively control this disease. Summary of the Invention
[0004] The purpose of this invention is to provide the application of glycoside alkaloid compounds in the prevention and / or treatment of bacterial leaf blight in plants, and to provide an inhibitor of bacterial leaf blight in rice, so as to achieve the effects of inhibiting the disease and improving disease resistance.
[0005] The application of glycoside alkaloids in the prevention and / or treatment of plant bacterial blight, wherein the glycoside alkaloids have one of the following chemical formulas:
[0006]
[0007] Another object of the present invention is to provide the application of glycoside alkaloid compounds or formulations containing glycoside alkaloid compounds as inhibitors of plant bacterial blight, wherein the glycoside alkaloid compounds have one of the following chemical structural formulas: .
[0008] The formulation containing glycoside alkaloids described in this invention uses solanine and / or lycopene as the active ingredient and also includes excipients. These glycoside alkaloids can be used alone or in combination with other excipients in the inhibition of rice bacterial blight.
[0009] The glycoside alkaloid compounds or preparations containing glycoside alkaloid compounds described in this invention have at least one of the following functions in inhibiting plant bacterial blight: Inhibit the expansion of rice bacterial blight lesions; It can alleviate the stress of bacterial blight in rice and increase the weight of rice roots and above-ground parts. It can alleviate the stress of bacterial blight in rice and improve the growth of rice roots and above-ground parts.
[0010] Furthermore, the plant in question is rice.
[0011] Furthermore, the glycoside alkaloids were extracted from Solanum nigrum (Latin name: *Solanum nigrum*). Solanum nigrum L.) green fruit.
[0012] Furthermore, the glycoside alkaloids are products obtained by column chromatography from the methanol extract of Solanum nigrum fruit, with chloroform:methanol (V / V) = 15:1 as the elution conditions. The resulting products include solanine and solanine alkaloid.
[0013] The products obtained by the above method include solanine and solanine, which can be used as active ingredients.
[0014] Another objective of this invention is to provide an inhibitor for rice bacterial blight.
[0015] An inhibitor for rice bacterial blight, wherein the active ingredient of the inhibitor is a glycoside alkaloid compound, and the glycoside alkaloid compound has one of the following chemical structural formulas:
[0016] Furthermore, the glycoside alkaloids in the inhibitor are products obtained by column chromatography from the methanol extract of Solanum nigrum fruit, with chloroform:methanol (V / V) = 15:1 as the elution conditions. The resulting products include solanine and solanine alkaloid.
[0017] Furthermore, the inhibitor has at least one of the following functions: Inhibit the expansion of rice bacterial blight lesions; It can alleviate the stress of bacterial blight in rice and increase the weight of rice roots and above-ground parts. It can alleviate the stress of bacterial blight in rice and improve the growth of rice roots and above-ground parts.
[0018] Furthermore, the inhibitor is composed of glycosidic alkaloid compounds and excipients.
[0019] Furthermore, the inhibitor, by mass percentage, comprises the following components: 0.5-20% glycosidic alkaloids, 0.5-2% cosolvent, 0.2-2% preservative, 1-10% excipients, and the balance being water.
[0020] The co-solvent is one or two of Tween 20, Tween 80, sodium dodecyl sulfate, sodium dodecyl sulfonate, and sodium heptane sulfonate; the preservative is one of sodium benzoate, benzoic acid, sorbic acid, potassium sorbate, sodium diacetate, sodium dehydroacetate, and butyl p-hydroxybenzoate; and the excipients are one of castor oil polyoxyethylene polyoxypropylene ether, alkylphenol polyoxyethylene ether formaldehyde condensate, glycerol polyoxyethylene ether fatty acid ester, fatty acid polyoxyethylene ester phosphate, fatty alcohol polyoxyethylene ether, and alkyl polyoxyethylene ether phosphate.
[0021] Another object of the present invention is to provide a method for suppressing bacterial blight in rice.
[0022] A method for suppressing rice bacterial blight involves spraying rice seedlings at the tillering stage with the aforementioned rice bacterial blight inhibitor, wherein the active ingredient of the inhibitor is a glycoside alkaloid compound, and the glycoside alkaloid compound has one of the following chemical structural formulas:
[0023] Furthermore, the application amount of the rice bacterial blight inhibitor, calculated as a glycoside alkaloid compound, is 50-200 μg / plant, with the optimal choice being 100 μg / plant.
[0024] The beneficial effects of this invention are as follows: Phenotypic observation and lesion area statistics show that the rice bacterial blight inhibitor of this invention has a significant inhibitory effect on leaf yellowing, lesions, and curling caused by pathogenic bacteria. This rice bacterial blight inhibitor can be applied to the leaves of rice seedlings during the tillering stage, thereby achieving the effects of disease suppression and improved disease resistance. This method is simple and inexpensive. The rice bacterial blight inhibitor described in this invention can significantly inhibit yellowing, lesions, and curling of rice leaves caused by pathogenic bacteria, thereby improving the disease resistance of rice. Attached Figure Description
[0025] Figure 1 Chromatographic comparison of glycoside alkaloid extract with standards of solanine and solanine alkaloid. Detailed Implementation
[0026] The following non-limiting embodiments are intended to enable those skilled in the art to more fully understand the invention, but do not limit the invention in any way.
[0027] Unless otherwise specified, the experimental methods described in the following examples are conventional methods; the reagents and materials described are commercially available unless otherwise specified.
[0028] Example 1: Inhibitory effect of different concentrations of solanine and lycopene on the pathogen causing bacterial blight of rice. First, the pathogen causing rice bacterial blight, Xanthomonas aeruginosa (… Xanthomonas oryzae pv. oryzae The bacteria were activated with Xoo, then inoculated in petri dishes using the streak method and incubated at 28 ℃ until clear single colonies appeared. A single colony was picked and inoculated into 4 mL of fresh PSB liquid medium, and cultured at 28 ℃ and 180 rpm for 24 h with shaking. After the bacterial suspension became noticeably turbid, 1 mL of the suspension was diluted 20 times with 19 mL of PSB liquid medium to prepare a bacterial suspension for later use. A certain amount of commercially available solanine and solanine oleoresin were accurately weighed using an electronic balance, dissolved in DMSO, and prepared into solutions with a concentration of 2048 μg / mL. Each well in the first column of a 96-well plate contained 1 μL of the stock solution of the compound and 199 μL of PSB liquid medium, and was thoroughly mixed by pipetting. For each column except the first column, add 100 μL of PSB liquid medium. Then, pipette 100 μL from the first column and add it to the second column, mixing thoroughly. Repeat this process until the desired concentration is achieved in the last column. Finally, add 100 μL of a 20-fold diluted bacterial suspension to each column to achieve a final test compound concentration of 512-16 μg / mL. The final volume for each well is 200 μL, and three biological replicates are set up for each concentration. Each 96-well plate includes a solvent control, a negative control, and a blank control, with three biological replicates for each control group.
[0029] The 96-well plate was incubated at 28 °C, and the OD value was measured every 12 hours. The OD value was read using a microplate reader. 600 The absorbance at the specified point was used. The inhibition rate was calculated according to the formula: inhibition rate (I%) = 100% × (OD value of solvent control - OD value of experimental group) / OD value of solvent control, and the half-maximum inhibitory concentration (IC50) was calculated using SPSS (27.0). 50 ).
[0030] The results showed that the inhibitory activity of solanine and lycoside against the pathogen of rice bacterial blight was concentration-dependent, with an IC50 value of 72 h. 50 The concentrations were 77.0 μg / mL and 64.6 μg / mL, respectively, indicating good antibacterial activity.
[0031] Table 1. Inhibition rate of different concentrations of solanine against rice bacterial blight pathogens
[0032] Table 2. Medium concentration of solanine inhibiting rice bacterial blight pathogen.
[0033] Table 3. Inhibition rate of different concentrations of lycorine on the pathogen of rice bacterial blight
[0034] Table 4. Medium concentration of lycorine inhibiting the pathogen of rice bacterial blight.
[0035] Isolation and identification of glycoside alkaloids used in Examples 2-6 below: Methanol extraction was performed on 20 kg of *Solanum nigrum* fruit for 24 h, repeated three times. The extract was concentrated using a rotary evaporator to obtain 600 g of methanol extract. The methanol extract was separated by silica gel column chromatography with chloroform:methanol (V / V) = 15:1 as the elution condition, yielding 20 g of glycosidic alkaloids (SGAs) (purity >90%). 1 mg of the SGAs compound was dissolved in 1 mL of chromatographic methanol, filtered through a 0.22 μm organic filter into a liquid chromatography vial to obtain the analyte. Qualitative and quantitative analysis was performed using HPLC. The determination was performed according to the high-performance liquid chromatography method (General Chapter 0512, Chinese Pharmacopoeia 2015 Edition), using octadecylsilane-bonded silica gel as the stationary phase and acetonitrile-0.3% phosphoric acid solution (containing 0.5% triethylamine) (V:V = 23:77) as the mobile phase, with a detection wavelength of 203 nm. Compared with the chromatogram of the standard, two glycoalkaloids, solanine and solanine, were detected in the SGAs sample, with solanine accounting for 44.8% and solanine accounting for 45.4%.
[0036] Example 2: Inhibitory effect of different concentrations of glycoside alkaloids on the pathogen causing bacterial blight in rice. A certain amount of glycoside alkaloids extracted from the green fruit of *Solanum nigrum* was accurately weighed using an electronic balance, dissolved in DMSO, and prepared into a solution with a concentration of 2048 μg / mL. The preparation method and 96-well plate operation of the suspension of *Xanthomonas xanthomonas*, the pathogen of rice bacterial blight, were the same as in Example 1. The 96-well plates were incubated at 28 °C, and the OD value was measured every 12 h. The OD values were read using a microplate reader. 600 The absorbance at the specified point. The inhibition rate (I%) was calculated using the formula: Inhibition rate (I%) = 100% × (OD value of solvent control - OD value of experimental group) / OD value of solvent control. The half-maximum inhibitory concentration (IC50) was then calculated using SPSS (27.0). 50 ).
[0037] The results showed that the glycoside alkaloids extracted from the green fruit of *Solanum nigrum* exhibited concentration-dependent inhibitory activity against the pathogen causing bacterial blight of rice, with an IC50 value of [value missing] at 72 h. 50 The concentration was 78.9 μg / mL, indicating good antibacterial activity.
[0038] Table 5. Inhibition rates of different concentrations of glycoside alkaloids against the pathogen causing bacterial blight of rice.
[0039] Table 6. Inhibitory concentrations of glycoside alkaloids against rice bacterial blight pathogens
[0040] Example 3: Inhibitory effect of different application amounts of glycoside alkaloids on rice bacterial blight The inhibitory effect of glycoalkaloids extracted from the green fruit of *Solanum nigrum* on yellowing of infected rice leaves under different application rates was tested using a hydroponic method. Before the experiment, rice seeds were vernalized at 4 ℃ for 2-3 days. After vernalization, the rice seeds were transferred to rice trays and cultured at 25 ℃ using Holland nutrient solution, maintaining a light intensity of 1200 Lx. Nutrient solution was replenished regularly until the tillering stage, at which point different treatments were applied. The blank control group was sprayed with only distilled water, while the experimental groups were sprayed with the pathogen and then with different concentrations of glycoalkaloid solutions dissolved in DMSO (0, 12.5, 25, 50, 100, 150, 200 μg / plant). On day 7, the area of lesions (cm²) on the rice leaves was measured. 2 )statistics.
[0041] The results showed that different amounts of glycoside alkaloids significantly inhibited the area of leaf lesions in rice, with the most significant inhibitory effect observed at an application rate of 100 μg / plant.
[0042] Table 7. Inhibitory activity of glycoside alkaloids against leaf tip lesions of rice bacterial blight
[0043] Example 4: Stress-relieving effects of different application amounts of glycoside alkaloids on rice infected with bacterial blight. The glycoside alkaloids extracted from the green fruit of *Solanum nigrum* were formulated into the application gradients described in Example 3, and the stress-relieving effects of the compounds on susceptible rice were tested using the hydroponic method of Example 3. The results showed that different application amounts of the glycoside alkaloids had a stress-relieving effect on rice infected with bacterial blight, exhibiting a certain concentration dependence, with the most significant relief effect observed at 100 μg / plant.
[0044] Table 8. Stress-relieving effects of glycoside alkaloids on root and aboveground part length in rice infected with bacterial leaf blight.
[0045] Table 9. Stress-relieving effects of glycoside alkaloids on root and aboveground weight of rice infected with bacterial leaf blight.
[0046] Example 5: Preparation of glycoside alkaloid compounds as an inhibitor of rice bacterial leaf blight The rice bacterial blight inhibitor, prepared from glycoalkaloid compounds extracted from the green fruit of Solanum nigrum, consists of the following components by mass percentage: 0.5-20% glycoalkaloids, 0.5-2% solubilizer, 0.2-2% preservative, 1-10% excipients and additives, and the balance being water.
[0047] The co-solvent is one or two of Tween 20, Tween 80, sodium dodecyl sulfate, sodium dodecyl sulfonate, and sodium heptane sulfonate; the preservative is one of sodium benzoate, benzoic acid, sorbic acid, potassium sorbate, sodium diacetate, sodium dehydroacetate, and butyl p-hydroxybenzoate; and the excipients are one of castor oil polyoxyethylene polyoxypropylene ether, alkylphenol polyoxyethylene ether formaldehyde condensate, glycerol polyoxyethylene ether fatty acid ester, fatty acid polyoxyethylene ester phosphate, fatty alcohol polyoxyethylene ether, and alkyl polyoxyethylene ether phosphate.
[0048] Formula 1: 5% glycoside alkaloids, 1% Tween 20, 1% sodium benzoate, 5% glycerol polyoxyethylene ether fatty acid ester, and the remainder water.
[0049] Formula 2: 10% glycoside alkaloids, 0.5% sodium dodecyl sulfonate, 0.2% sodium benzoate, 10% alkylphenol polyoxyethylene ether formaldehyde condensate, and the balance being water.
[0050] Formula 3: 0.5% glycoside alkaloids, 2% sodium dodecyl sulfate, 2% potassium sorbate, 1% castor oil polyoxyethylene polyoxypropylene ether, and the remainder water.
[0051] Formula 4: 8% glycoside alkaloids, 1% Tween 80, 0.8% sodium benzoate, 1% fatty alcohol polyoxyethylene ether, and the remainder water.
[0052] Formula 5: 10% glycoside alkaloids, 1% sodium heptanesulfonate, 0.5% sodium benzoate, 4% castor oil polyoxyethylene polyoxypropylene ether, and the remainder water.
[0053] Formula 6: 10% glycoside alkaloids, 0.5% sodium dodecyl sulfonate, 0.5% Tween 20, 0.5% sodium benzoate, 2% alkyl polyoxyethylene ether phosphate, and the remainder water.
[0054] Example 6: Application of glycoside alkaloids as inhibitors of rice bacterial leaf blight in suppressing rice bacterial leaf blight Glycoside alkaloid rice bacterial blight inhibitors were obtained according to formulations 1-6 in Example 5. Nipponbare rice seeds were germinated, and then seedlings were transplanted to experimental fields. The glycoside alkaloid rice bacterial blight inhibitors (formulas 1-6) were sprayed at a dosage of 30 g / mu (approximately 0.5 mg inhibitor / plant), with a control group (sprayed only with pathogenic bacteria solution). Twenty days after spraying, 10 seedlings from each control group were selected, and the lesion area (square centimeters, cm²) was calculated. 2 ), length (cm) and weight (g) of roots and above-ground parts.
[0055] Table 10. Inhibitory effects of glycoside alkaloids on rice bacterial blight.
[0056] The experimental results above show that the compound glycoside alkaloid of the present invention can significantly inhibit rice bacterial blight.
Claims
1. The application of glycoside alkaloid compounds in the prevention and / or treatment of plant bacterial blight, characterized in that: The glycoside alkaloid compound has one of the following chemical structural formulas: 。 2. The application of glycoside alkaloid compounds or preparations containing glycoside alkaloid compounds as inhibitors of plant bacterial blight, characterized in that: The glycoside alkaloid compound has one of the following chemical structural formulas: 。 3. The application according to claim 1 or 2, characterized in that: The plant in question is rice.
4. The application according to claim 1 or 2, characterized in that: The glycoalkaloid compounds are products obtained by column chromatography from the methanol extract of Solanum nigrum fruit, with chloroform:methanol (V:V) = 15:1 as the elution conditions. The obtained products include solanine and solanine alkaloid.
5. A rice bacterial blight inhibitor, characterized in that: The active ingredient of the inhibitor is a glycoside alkaloid compound, and the glycoside alkaloid compound has one of the following chemical structural formulas: 。 6. The inhibitor according to claim 1, characterized in that: The inhibitor has at least one of the following functions: Inhibit the expansion of rice bacterial blight lesions; It can alleviate the stress of bacterial blight in rice and increase the weight of rice roots and above-ground parts. It can alleviate the stress of bacterial blight in rice and improve the growth of rice roots and above-ground parts.
7. The inhibitor according to claim 1, characterized in that: The inhibitor is composed of glycoside alkaloids and excipients.
8. The inhibitor according to claim 1, characterized in that: The inhibitor, by mass percentage, comprises the following components: 0.5-20% glycoside alkaloids, 0.5-2% solubilizer, 0.2-2% preservative, 1-10% excipients, and the balance being water. The co-solvent is one or two of Tween 20, Tween 80, sodium dodecyl sulfate, sodium dodecyl sulfonate, and sodium heptane sulfonate; the preservative is one of sodium benzoate, benzoic acid, sorbic acid, potassium sorbate, sodium diacetate, sodium dehydroacetate, and butyl p-hydroxybenzoate; and the excipients are one of castor oil polyoxyethylene polyoxypropylene ether, alkylphenol polyoxyethylene ether formaldehyde condensate, glycerol polyoxyethylene ether fatty acid ester, fatty acid polyoxyethylene ester phosphate, fatty alcohol polyoxyethylene ether, and alkyl polyoxyethylene ether phosphate.
9. A method for suppressing bacterial leaf blight in rice, characterized in that: Spray rice seedlings during the tillering stage with the inhibitor of rice bacterial blight as described in claim 4.
10. The method according to claim 9, characterized in that: The dosage of the rice bacterial blight inhibitor, calculated as glycoside alkaloid compounds, is 50-200 μg / plant.