A pharmaceutical composition for preventing and treating rice panicle blight

Abstract

The application discloses a medicine composition for preventing and treating rice panicle blight, and relates to the technical field of rice planting.The medicine composition for preventing and treating rice panicle blight is prepared from the following raw materials in parts by mass: cycas revoluta extract 15-20 parts and citrus aurantium fruit polysaccharide 12-15 parts.The medicine composition of the application is prepared from natural plant extracts as raw material components, and thus environmental hazards caused by chemical reagents are avoided, and the medicine composition has the characteristics of green environmental protection; meanwhile, the medicine composition of the application is prepared from two kinds of raw materials, and thus the component collocation is simpler.The medicine composition of the application has a remarkable effect on preventing and treating rice panicle blight, and has important positive significance for promoting the healthy and sustainable development of the rice industry.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of rice planting, in particular to a medicine composition for preventing and treating rice panicle blight. BACKGROUND

[0002] Rice panicle blight is mainly caused by Burkholderia glumae, and the disease is characterized by "green stem, green leaves and grain blight". The disease occurs in the late growth period of rice and can cause serious yield reduction, resulting in huge economic losses to agricultural production.

[0003] At present, the prevention and treatment of rice panicle blight mainly includes agricultural prevention and chemical prevention. The agricultural prevention methods mainly include seed disinfection, delayed sowing to avoid the flowering period of rice and high temperature weather, and improvement of water and fertilizer management. As for chemical prevention, there is no mature commercial panicle blight prevention agent. Ammonium dithane is commonly used as a prevention agent, or 75% naodiwen+46% CU(OH)2, 12.5% enazole+25% prochloraz, 10% benzene ether azole+33.5% quinoline copper complex are used for the prevention of rice smut and panicle blight, but the prevention effect is not ideal.

[0004] With the global climate change and the change of cultivation methods, the incidence of rice panicle blight is increasing, which has a very bad impact on rice planting. At the same time, the toxin produced by the pathogen of panicle blight can further affect human and animal health. Therefore, it is necessary to effectively prevent and treat rice panicle blight, which has important positive significance for promoting the healthy development of agriculture. SUMMARY

[0005] The present application aims to provide a medicine composition for preventing and treating rice panicle blight, so as to solve the problems existing in the prior art and achieve effective prevention and treatment of rice panicle blight.

[0006] To achieve the above-mentioned purpose, the present application provides the following solutions.

[0007] The present application provides a medicine composition for preventing and treating rice panicle blight, and the raw material mass ratio of the medicine composition is as follows:

[0008] 15-20 parts of cycas revoluta extract and 12-15 parts of citrus aurantium polysaccharide.

[0009] Further, the extraction method of the cycas revoluta extract in the present application comprises the following steps:

[0010] The leaf part of the cycas is decocted, the decocted liquid is concentrated, the alcohol content of the decocted liquid is adjusted to 45-50% by using anhydrous ethanol, the precipitate is discarded, then anhydrous ethanol is continuously added, the alcohol content is adjusted to 70-75%, and the precipitate is collected and dried to obtain the cycas extract.

[0011] Further, the extraction method of the Citrus maxima fruit polysaccharide in the application comprises the following steps:

[0012] The Citrus maxima fruit is fermented by using Bacillus amyloliquefaciens to obtain a fermentation liquid;

[0013] The fermentation liquid is concentrated under reduced pressure, then ethanol is added, and the crude extract is obtained by centrifugal drying;

[0014] The crude extract is dissolved in deionized water, the protein is removed by using the Sevag method, the obtained solution is concentrated under reduced pressure, then anhydrous ethanol is added to make the alcohol content of the system 55-65%, the precipitate is collected and dried to obtain the Citrus maxima fruit polysaccharide.

[0015] As a further preferred embodiment of the application, the fermentation temperature is 37 DEG C, and the fermentation time is 3-4 days.

[0016] The alcohol extraction method can separate many kinds of phytochemical components, such as alkaloids, terpenes, steroids and flavonoids. The cycas leaf part is subjected to alcohol extraction in the application, and the polysaccharide material extracted after fermentation of the active material and the Citrus maxima fruit plays a significant effect in preventing and treating rice panicle blight.

[0017] The application also provides the use of the above-mentioned pharmaceutical composition in preventing and treating rice panicle blight.

[0018] The application further provides a medicament for preventing and treating rice panicle blight, which takes the cycas extract and the Citrus maxima fruit polysaccharide as active ingredients.

[0019] As a further preferred embodiment of the application, the mass ratio of the cycas extract and the Citrus maxima fruit polysaccharide is 15-20:12-15.

[0020] The application discloses the following technical effects:

[0021] The pharmaceutical composition of the application takes natural plant extracts as raw material components, avoids environmental hazards caused by chemical reagents, and has the characteristics of green environmental protection; meanwhile, the pharmaceutical composition of the application adopts two kinds of raw materials, and the component collocation is simpler.

[0022] The pharmaceutical composition of the application has a significant effect in preventing and treating rice panicle blight, and has important positive significance for promoting the healthy and sustainable development of the rice industry. DETAILED DESCRIPTION

[0023] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0024] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0025] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0026] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be readily apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0027] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0028] Example 1

[0029] The drug composition for controlling rice panicle blight consists of the following raw materials in the indicated weight ratios:

[0030] 15 parts of cycad extract and 14 parts of citron fruit polysaccharide;

[0031] Preparation of cycad extract:

[0032] The leaves of the cycad were crushed, and decocted three times with 8 times their weight of water for 45 minutes each time. The decoction was collected. The decoction was then concentrated to 50% of its original volume, and anhydrous ethanol was added to make the alcohol content of the system 50%. After standing, the system was centrifuged and the precipitate was discarded. Anhydrous ethanol was then added to make the alcohol content 70%. After standing, the system was filtered, the precipitate was collected, and dried to obtain the cycad extract.

[0033] Extraction of polysaccharides from Citrus medica fruits:

[0034] a. After crushing the Citrus medica fruits, Bacillus amyloliquefaciens (purchased from China General Microbiological Culture Collection Center, preservation number CGMCC No. 17841) was added, and fermentation was carried out at 37°C for 3 days to obtain a fermentation liquor;

[0035] b. After the fermentation liquor obtained in step a was concentrated under reduced pressure, ethanol was added, and centrifugal drying was carried out to obtain a crude extract;

[0036] c. The crude extract obtained in step b was dissolved in deionized water, and the Sevag method was used to remove proteins (chloroform:amyl alcohol 4:1 mixed), to obtain a solution. After concentration under reduced pressure, chloroform and amyl alcohol were removed, and then anhydrous ethanol was added to make the alcohol content of the system 60%, and filtration was carried out. The precipitate was collected and freeze-dried to obtain polysaccharides from Citrus medica fruits.

[0037] Example 2

[0038] The pharmaceutical composition for preventing and treating rice panicle blight is composed of the following raw materials in parts by mass:

[0039] 20 parts of cycas revoluta extract and 15 parts of polysaccharides from Citrus medica fruits;

[0040] Preparation of cycas revoluta extract:

[0041] The leaf part of cycas revoluta was crushed, 8 times the mass of water was added, and decoction was carried out 3 times, each time for 45 min, and the decoction liquid was collected. Then the decoction liquid was concentrated to 50% of the original volume, anhydrous ethanol was added to make the alcohol content of the system 45%, and after standing, centrifugal separation was carried out, and the precipitate part was discarded. Then anhydrous ethanol was continuously added to make the alcohol content 70%, and after standing, suction filtration was carried out, the precipitate was collected, and after drying, cycas revoluta extract was obtained.

[0042] Extraction of polysaccharides from Citrus medica fruits:

[0043] a. After crushing the Citrus medica fruits, Bacillus amyloliquefaciens (purchased from China General Microbiological Culture Collection Center, preservation number CGMCC No. 17841) was added, and fermentation was carried out at 37°C for 3 days to obtain a fermentation liquor;

[0044] b. After the fermentation liquor obtained in step a was concentrated under reduced pressure, ethanol was added, and centrifugal drying was carried out to obtain a crude extract;

[0045] c. The crude extract obtained in step b was dissolved in deionized water, and the Sevag method was used to remove proteins (chloroform:amyl alcohol 4:1 mixed), to obtain a solution. After concentration under reduced pressure, chloroform and amyl alcohol were removed, and then anhydrous ethanol was added to make the alcohol content of the system 55%, and filtration was carried out. The precipitate was collected and freeze-dried to obtain polysaccharides from Citrus medica fruits.

[0046] Example 3

[0047] The medicine composition for preventing and treating rice panicle blight is composed of the following raw materials in mass parts:

[0048] 16 parts of Cycas revoluta extract and 12 parts of Citrus aurantium polysaccharide;

[0049] Preparation of Cycas revoluta extract:

[0050] The leaf part of Cycas revoluta is crushed, 8 times of water by mass is added and decocted for 3 times, each time for 45 min, and the decocted liquid is collected; then the decocted liquid is concentrated to 50% of the original volume, anhydrous ethanol is added to make the alcohol content of the system 50%, after standing, centrifugation is performed, and the precipitate part is discarded; then anhydrous ethanol is continuously added to make the alcohol content 75%, after standing, suction filtration is performed, the precipitate is collected, and after drying, Cycas revoluta extract is obtained.

[0051] Extraction of Citrus aurantium polysaccharide:

[0052] a. After the Citrus aurantium fruit is crushed, Bacillus amyloliquefaciens (purchased from China General Microbiological Culture Collection Center, preservation number CGMCC No. 17841) is added, and fermentation is carried out at 37°C for 3 days to obtain a fermentation liquid;

[0053] b. After the fermentation liquid obtained in step a is reduced pressure concentrated, ethanol is added, centrifugal drying is performed to obtain a crude extract;

[0054] c. The crude extract obtained in step b is dissolved in deionized water, protein is removed by Sevag method (chloroform:amyl alcohol 4:1 mixed), a solution is obtained, after reduced pressure concentration, chloroform and amyl alcohol are removed, then anhydrous ethanol is added to make the alcohol content of the system 60%, filtration is performed, the precipitate is collected, and freeze-drying is performed to obtain Citrus aurantium polysaccharide.

[0055] Example 4

[0056] The medicine composition for preventing and treating rice panicle blight is composed of the following raw materials in mass parts:

[0057] 18 parts of Cycas revoluta extract and 13 parts of Citrus aurantium polysaccharide;

[0058] Preparation of Cycas revoluta extract:

[0059] The leaf part of Cycas revoluta is crushed, 8 times of water by mass is added and decocted for 3 times, each time for 45 min, and the decocted liquid is collected; then the decocted liquid is concentrated to 50% of the original volume, anhydrous ethanol is added to make the alcohol content of the system 45%, after standing, centrifugation is performed, and the precipitate part is discarded; then anhydrous ethanol is continuously added to make the alcohol content 70%, after standing, suction filtration is performed, the precipitate is collected, and after drying, Cycas revoluta extract is obtained.

[0060] Extraction of Citrus aurantium polysaccharide:

[0061] a. crushing the fruit of Citrus medica L. and adding Bacillus amyloliquefaciens (purchased from China General Microbiological Culture Collection Center, with the preservation number of CGMCC No. 17841), and then fermenting at 37°C for 3 days to obtain a fermentation liquor;

[0062] b. adding ethanol to the fermentation liquor obtained in step a. after concentration under reduced pressure, and then centrifuging and drying to obtain a crude extract;

[0063] c. dissolving the crude extract obtained in step b. in deionized water, removing proteins by the Sevag method (chloroform:amyl alcohol 4:1 mixed), to obtain a solution, and then removing chloroform and amyl alcohol after concentration under reduced pressure, and then adding anhydrous ethanol to make the alcohol content of the system 55%, filtering, collecting the precipitate, and freeze-drying to obtain the polysaccharide of the fruit of Citrus medica L.

[0064] Comparative Example 1

[0065] The difference from Example 1 is only that step a. of the extraction process of the polysaccharide of the fruit of Citrus medica L. adopts a decocting method:

[0066] a. crushing the fruit of Citrus medica L. and adding 10 times the mass of water to decoct 3 times, each time for 45 min, and collecting the decocting liquor;

[0067] The remaining steps are the same as those in Example 1.

[0068] Comparative Example 2

[0069] The difference from Example 1 is only that the leaf part of Cycas revoluta Thunb. is replaced by the same mass of Cycas revoluta Thunb. flowers to extract.

[0070] Effect verification example:

[0071] 1. Bacteriostatic performance test

[0072] A bacterial suspension of the strain LMG2196 of Burkholderia glumae with a concentration of 3 10 8 cfu / mL was mixed with sterilized NA medium according to a volume ratio of 1:15, and then placed in a culture dish for bacteriostatic test; the pharmaceutical compositions of Examples 1-4 and Comparative Examples 1-2 were respectively dissolved in sterile water to different concentrations, and then 50 μL of each was dropped on a sterile filter paper sheet in the culture dish to perform a bacteriostatic circle test.

[0073] Each test was repeated 3 times, and the average value was calculated, and the results are shown in Table 1.

[0074] Table 1

[0075]

[0076] As can be seen from Table 1, the pharmaceutical composition of the present application has a significant inhibitory effect on the pathogenic bacteria of rice panicle blast disease, and has a bacteriostatic effect basically equivalent to that of the chemical agent metam.

[0077] 2. Field control experiment

[0078] (1) Experimental site: a rice test field in Jiangxi Province.

[0079] (2) Experimental grouping: 8 equal-area plots were selected, each with an area of 30 m 2 (10 m 3 m), which were respectively used as treatment groups 1-4, control groups 1-2, a positive control group, and a blank control group.

[0080] (3) Experimental treatment: After the rice seeds were sown, each group was sprayed with a bacterial suspension of the sheath bacteria Burkholderia glumae strain LMG2196 at a concentration of 3 10 8 cfu / mL on the 3rd day of the heading stage of the rice, until the droplets were dripping from the leaf surface, and then the rice was tied with a rope to keep it moist for 3 days to allow the bacteria to invade.

[0081] On the 6th day after spraying the bacterial suspension, treatment groups 1-4 and control groups 1-2 were sprayed with 500-fold diluted solutions of the pharmaceutical compositions of Examples 1-4 and Comparative Examples 1-2, respectively, on the rice panicles, with 2 sprays during the growth period and a 7-day interval; the blank control group was sprayed with an equal amount of water, and the positive control group was sprayed with an equal amount of 45% ammonium sulfate 1000-fold diluted solution.

[0082] (4) Field investigation: using the five-point sampling method, the rice panicle blast disease was investigated in the field, with 12 panicles randomly taken from each point, and the percentage of diseased grains and the disease grade were recorded.

[0083] Among them, the panicle blast disease grading standards are as follows:

[0084] 0 level: rice panicle without discoloration; 1 level: 1-10% of the grains discolored; 2 level: 11-20% of the grains discolored; 3 level: 21-30% of the grains discolored; 4 level: 31-40% of the grains discolored; 5 level: 41-50% of the grains discolored; 6 level: 51-60% of the grains discolored; 7 level: 61-70% of the grains discolored; 8 level: 71-80% of the grains discolored; 9 level: > 81% of the grains discolored.

[0085] The calculation formula of the disease index is as follows:

[0086] ;

[0087] The calculation formula of the control effect is as follows:

[0088] ;

[0089] Among them, Y is the control effect; r T is the disease performance growth rate of the treatment group; r ck- the rate of increase in disease expression of the control group.

[0090] (5) Result statistics: the experimental results are shown in Table 2.

[0091] Table 2

[0092]

[0093] The above-described embodiments are merely preferred modes of the present application and are not intended to limit the scope of the present application. Various modifications and improvements to the technical solutions of the present application made by those of ordinary skill in the art without departing from the design spirit of the present application shall fall within the scope of protection of the present application as defined by the claims.

Claims

1. A pharmaceutical composition for controlling rice panicle blight, characterized in that, The raw material mass ratio is: 15-20 parts of the cycas extract and 12-15 parts of the citrus medica fruit polysaccharide; The extraction method of the cycas extract comprises the following steps: The cycas leaves are decocted, and the decocted liquid is concentrated; the alcohol content of the decocted liquid is adjusted to 45-50% by using anhydrous ethanol; after the precipitate is discarded, anhydrous ethanol is continuously added to adjust the alcohol content to 70-75%; the system is left to stand, filtered, and the precipitate is collected and dried to obtain the cycas extract; The extraction method of the citrus medica fruit polysaccharide comprises the following steps: The citrus medica fruit is fermented by using bacillus amyloliquefaciens to obtain a fermentation liquid; After the fermentation liquid is concentrated under reduced pressure, ethanol is added, and the crude extract is obtained by centrifugal drying; The crude extract is dissolved in deionized water, and the protein is removed by using the Sevag method; the obtained solution is concentrated under reduced pressure, and then anhydrous ethanol is added to adjust the alcohol content of the system to 55-65%; the system is filtered, and the precipitate is collected and dried to obtain the citrus medica fruit polysaccharide; The fermentation temperature is 37 DEG C, and the fermentation time is 3-4 days.

2. The application of the pharmaceutical composition in claim 1 in preventing and treating rice panicle blight.

3. An agent for controlling rice panicle blight, characterized by, The cycas extract and the citrus medica fruit polysaccharide are used as active ingredients; The extraction method of the cycas extract comprises the following steps: The cycas leaves are decocted, and the decocted liquid is concentrated; the alcohol content of the decocted liquid is adjusted to 45-50% by using anhydrous ethanol; after the precipitate is discarded, anhydrous ethanol is continuously added to adjust the alcohol content to 70-75%; the system is left to stand, filtered, and the precipitate is collected and dried to obtain the cycas extract; The extraction method of the citrus medica fruit polysaccharide comprises the following steps: The citrus medica fruit is fermented by using bacillus amyloliquefaciens to obtain a fermentation liquid; After the fermentation liquid is concentrated under reduced pressure, ethanol is added, and the crude extract is obtained by centrifugal drying; The crude extract is dissolved in deionized water, and the protein is removed by using the Sevag method; the obtained solution is concentrated under reduced pressure, and then anhydrous ethanol is added to adjust the alcohol content of the system to 55-65%; the system is filtered, and the precipitate is collected and dried to obtain the citrus medica fruit polysaccharide; The fermentation temperature is 37 DEG C, and the fermentation time is 3-4 days.

4. The medicament according to claim 3, characterized in that, The mass ratio of the cycas extract and the citrus medica fruit polysaccharide is 15-20:12-15.

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