CN104026155A - Preparation method of biological source mixed bactericide - Google Patents
Preparation method of biological source mixed bactericide Download PDFInfo
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- CN104026155A CN104026155A CN201410295505.5A CN201410295505A CN104026155A CN 104026155 A CN104026155 A CN 104026155A CN 201410295505 A CN201410295505 A CN 201410295505A CN 104026155 A CN104026155 A CN 104026155A
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Abstract
The invention discloses a preparation method of biological source mixed bactericide. The preparation method comprises the following steps: (1) uniformly mixing auricularia auricula-judae sporocarp dry powder and water in a weight ratio of 1 to (40-50), extracting in water bath at 70-100 DEG C for 2-5 hours, filtering, centrifuging at the speed of 10000rpm for 20-50 minutes, performing alcohol precipitating on centrifugate by 70% ethanol, centrifuging at the speed of 8000-12000 rpm for 30 minutes, collecting precipitates, and drying to obtain an auricularia auricula-judae stabilizer; (2) uniformly mixing lentinula edodes sporocarp dry powder and water in a weight ratio of 1 to (15-30), digesting at 4 DEG C for 3-5 hours, centrifuging at the speed of 10000rpm for 25 minutes, precipitating supernatant by an ammonium sulfate precipitation with saturation of 40%-60%, standing in a refrigerator at 4 DEG C for 12 hours, centrifuging at the speed of 10000-12000 rpm for 15-30 minutes, collecting precipitates, placing in a dialysis bag, dialyzing by distilled water for 20-24 hours, collecting dialysate, and drying in vacuum to obtain a lentinus edodes bactericide; and (3) uniformly mixing the auricularia auricula-judae stabilizer and the lentinus edodes bactericide in a weight ratio of 1 to (8-12), and uniformly mixing the obtained mixture with water in a weight ratio of 1 to (30-40) to obtain the mixed bactericide. The preparation method disclosed by the invention is simple in process and strong in controllability, the bactericide has the rate of 53.55%-78.62% of inhibiting the growth of Colletotrichum orbiculare Arx and the rate of 80.43%-95.71% of inhibiting the growth of Phytophthora capsici, and the pesticide effect can last for 4-20 days and 8-12 days.
Description
Technical field
The invention belongs to the extraction of biopesticide in natural resources, particularly the extraction of bactericide.
Technical background
Watermelon is the good merchantable brand that quenches one's thirst of relieving summer heat Chinese traditional summer, watermelon anthrax (
colletotrichum orbicularearx) and Phytophthora capsici (
phytophthora capsici) two kinds of disease funguses can infect watermelon and directly affect its output and quality, and after infection process latent period short, rapid onset, prevents and treats very difficult.In order effectively to control the harm of these diseases, use at present maximum still chemical prevention and control methods.Because pathogen very easily develops immunity to drugs to bactericide, the effect of chemical control usually reduces greatly because of the drug-fast raising of germ.And the use of a large amount of chemical pesticides, give ecology, agricultural and environment have brought very large negative influence.
The biostatic agent that uses microorganism or animals and plants to make is to control one of safer and effective method of plant harmful fungoid, and researcher has carried out a large amount of research to this both at home and abroad, and obtains the extract of multiple tool potentiality to be exploited.20th century 60, the seventies, Japanese scholars suppresses activity to the phytopathogen of fungi extracts and conducts extensive research, and in 70, the eighties more separated successively have the active substance of bacteriostatic activity, some is for the production of in reality.Because natural antifungal active substance exists drug effect unstable in practice, the problems such as easy loss of activity, have limited the further exploitation of these biopesticides.
Summary of the invention
Order of the present invention ground is for two kinds of disease funguses on watermelon, and a kind of method that obtains mixed bactericide is provided.Can effectively solve these two problems of environmental protection and pharmacodynamic stability.
The present invention is achieved by the following technical solutions.
Preparation method of the present invention, according to the following steps.
(1) woodear fruit body dry powder and water are mixed by weight the ratio of l:40-50, extracting 2-5h in 70-100 ℃ of water-bath, gauze tentatively filters, the centrifugal 20-50min of 10000rpm, centrifugate 70% alcohol chromatography, the centrifugal 30min of 8000-12000rpm, the dry woodear stabilizing agent that obtains after collecting precipitation.
(2) mushroom fruiting body dry powder and water are mixed by weight the ratio of 1:15-30,4 ℃ of lixiviate 3-5h, and the centrifugal 25min of 10000rpm, collects supernatant, the ammonium sulfate ((NH of 40%-60% degree of saturation for supernatant
4)
2sO
4) precipitation, standing 12h in 4 ℃ of refrigerators, the centrifugal 15-30min of 10000-12000rpm, puts in bag filter and uses distill water dialysis 20-24h after precipitation is collected, and after the vacuum drying of collection dislysate, obtains mushroom bactericide.
(3) after the woodear stabilizing agent of step (1) and step (2) and mushroom bactericide are mixed by weight the ratio of 1:8-12, then be mixed with the ratio of water by weight 1:30-40, obtain mixed bactericide.
The present invention adopts growth rate method to detect the inhibitory action of bactericide to pathogen mycelial growth.Mixed bactericide reaches 87.85-100% to watermelon anthrax and Phytophthora capsici mycelia growth inhibition ratio, and duration of efficacy reaches 15-30 days.
Optimum condition of the present invention is according to the following steps.
(1) woodear fruit body dry powder and water are mixed by weight the ratio of l:40, extracting 4h in 80 ℃ of water-baths, and gauze tentatively filters, the centrifugal 20min of 10000rpm, centrifugate 70% alcohol chromatography, the centrifugal 30min of 10000rpm, the dry woodear stabilizing agent that obtains after collecting precipitation.
(2) mushroom fruiting body dry powder and water are mixed by weight the ratio of 1:20,4 ℃ of lixiviate 4h, the centrifugal 25min of 10000rpm, collect supernatant, the ammonium sulfate precipitation of 40%-60% degree of saturation for supernatant, standing 12h in 4 ℃ of refrigerators, the centrifugal 25min of 12000rpm, after precipitation is collected, put in bag filter and use distill water dialysis 24h, after the vacuum drying of collection dislysate, obtain mushroom bactericide.
(3) after the woodear stabilizing agent of step (1) and step (2) and mushroom bactericide are mixed by weight the ratio of 1:9, then be mixed with the ratio of water by weight 1:33, obtain mixed bactericide.
Technique of the present invention is simple, and controlled row is strong, and watermelon anthrax and Phytophthora capsici mycelia growth inhibition ratio are respectively to 53.55-78.62% and 80.43-95.71%, and duration of efficacy is 14-20 days and 8-12 days.Prepared mixed bactericide can be used for biopesticide exploitation.
Embodiment
The present invention will be described further by following examples.
Embodiment 1.
The weight ratio of woodear fruit body dry powder and water is l:50, extracting 3h in 100 ℃ of water-baths, and gauze tentatively filters, the centrifugal 40min of 10000rpm, centrifugate 70% alcohol chromatography, the centrifugal 30min of 12000rpm, the dry woodear stabilizing agent that obtains after collecting precipitation.
The weight ratio of mushroom fruiting body dry powder and water is 1:15,4 ℃ of lixiviate 5h, the centrifugal 25min of 10000rpm, collect supernatant, the ammonium sulfate precipitation of 40%-60% degree of saturation for supernatant, standing 12h in 4 ℃ of refrigerators, the centrifugal 30min of 10000rpm, after precipitation is collected, put in bag filter and use distill water dialysis 20h, after the vacuum drying of collection dislysate, obtain mushroom bactericide.
The weight ratio that woodear stabilizing agent and mushroom bactericide powder mix rear and water by the weight ratio of 1:8 is that 1:30 obtains mixed bactericide.
Adopt growth rate method to detect the inhibitory action of bactericide to pathogen mycelial growth.Mixed bactericide reaches 94.64% to watermelon anthrax and Phytophthora capsici mycelia growth inhibition ratio, and duration of efficacy reaches 28 days.
Embodiment 2.
The weight ratio of woodear fruit body dry powder and water is l:40, extracting 2h in 70 ℃ of water-baths, and gauze tentatively filters, the centrifugal 20min of 10000rpm, centrifugate 70% alcohol chromatography, the centrifugal 30min of 10000rpm, the dry woodear stabilizing agent that obtains after collecting precipitation.
The weight ratio of mushroom fruiting body dry powder and water is 1:30,4 ℃ of lixiviate 5h, the centrifugal 25min of 10000rpm, collect supernatant, the ammonium sulfate precipitation of 40%-60% degree of saturation for supernatant, standing 12h in 4 ℃ of refrigerators, the centrifugal 20min of 10000rpm, after precipitation is collected, put in bag filter and use distill water dialysis 20h, after the vacuum drying of collection dislysate, obtain mushroom bactericide.
The weight ratio that woodear stabilizing agent and mushroom bactericide powder mix rear and water by the weight ratio of 1:12 is that 1:35 obtains mixed bactericide.
Adopt growth rate method to detect the inhibitory action of bactericide to pathogen mycelial growth.Mixed bactericide reaches 90.25% to watermelon anthrax and Phytophthora capsici mycelia growth inhibition ratio, and duration of efficacy reaches 23 days.
Embodiment 3.
The weight ratio of woodear fruit body dry powder and water is l:45, extracting 5h in 80 ℃ of water-baths, and gauze tentatively filters, the centrifugal 50min of 10000rpm, centrifugate 70% alcohol chromatography, the centrifugal 30min of 8000rpm, the dry woodear stabilizing agent that obtains after collecting precipitation.
The weight ratio of mushroom fruiting body dry powder and water is 1:20,4 ℃ of lixiviate 3h, the centrifugal 25min of 10000rpm, collect supernatant, the ammonium sulfate precipitation of 40%-60% degree of saturation for supernatant, standing 12h in 4 ℃ of refrigerators, the centrifugal 15min of 12000rpm, after precipitation is collected, put in bag filter and use distill water dialysis 24h, after the vacuum drying of collection dislysate, obtain mushroom bactericide.
The weight ratio that woodear stabilizing agent and mushroom bactericide powder mix rear and water by the weight ratio of 1:10 is that 1:40 obtains mixed bactericide.
Adopt growth rate method to detect the inhibitory action of bactericide to pathogen mycelial growth.Mixed bactericide reaches 92.46% to watermelon anthrax and Phytophthora capsici mycelia growth inhibition ratio, and duration of efficacy reaches 27 days.
Claims (2)
1. a preparation method for biogenic mixed bactericide, is characterized in that according to the following steps:
(1) woodear fruit body dry powder and water are mixed by weight the ratio of l:40-50, extracting 2-5h in 70-100 ℃ of water-bath, gauze tentatively filters, the centrifugal 20-50min of 10000rpm, centrifugate 70% alcohol chromatography, the centrifugal 30min of 8000-12000rpm, the dry woodear stabilizing agent that obtains after collecting precipitation;
(2) mushroom fruiting body dry powder and water are mixed by weight the ratio of 1:15-30,4 ℃ of lixiviate 3-5h, the centrifugal 25min of 10000rpm, collect supernatant, the ammonium sulfate precipitation of 40%-60% degree of saturation for supernatant, standing 12h in 4 ℃ of refrigerators, the centrifugal 15-30min of 10000-12000rpm, after precipitation is collected, put in bag filter and use distill water dialysis 20-24h, after the vacuum drying of collection dislysate, obtain mushroom bactericide;
(3) after the woodear stabilizing agent of step (1) and step (2) and mushroom bactericide are mixed by weight the ratio of 1:8-12, then be mixed with the ratio of water by weight 1:30-40, obtain mixed bactericide.
2. preparation method according to claim 1, is characterized in that according to the following steps:
(1) woodear fruit body dry powder and water are mixed by weight the ratio of l:40, extracting 4h in 80 ℃ of water-baths, and gauze tentatively filters, the centrifugal 20min of 10000rpm, centrifugate 70% alcohol chromatography, the centrifugal 30min of 10000rpm, the dry woodear stabilizing agent that obtains after collecting precipitation;
(2) mushroom fruiting body dry powder and water are mixed by weight the ratio of 1:20,4 ℃ of lixiviate 4h, the centrifugal 25min of 10000rpm, collect supernatant, the ammonium sulfate precipitation of 40%-60% degree of saturation for supernatant, standing 12h in 4 ℃ of refrigerators, the centrifugal 25min of 12000rpm, after precipitation is collected, put in bag filter and use distill water dialysis 24h, after the vacuum drying of collection dislysate, obtain mushroom bactericide;
(3) after the woodear stabilizing agent of step (1) and step (2) and mushroom bactericide are mixed by weight the ratio of 1:9, then be mixed with the ratio of water by weight 1:33, obtain mixed bactericide.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20070009341A (en) * | 2005-07-15 | 2007-01-18 | 강원도 | Antifungal material for plant powdery mildew containing of tagetes patula or lentinus edodes |
| JP2011140463A (en) * | 2010-01-07 | 2011-07-21 | Tottori Univ | Technique for controlling plant disease using waste mushroom bed of edible mushroom |
| CN103461471A (en) * | 2013-09-04 | 2013-12-25 | 福建省农业科学院食用菌研究所 | Application of hypsizigus marmoreus lectin and deproteinization polysaccharide in preservation of loquats |
-
2014
- 2014-06-27 CN CN201410295505.5A patent/CN104026155B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20070009341A (en) * | 2005-07-15 | 2007-01-18 | 강원도 | Antifungal material for plant powdery mildew containing of tagetes patula or lentinus edodes |
| JP2011140463A (en) * | 2010-01-07 | 2011-07-21 | Tottori Univ | Technique for controlling plant disease using waste mushroom bed of edible mushroom |
| CN103461471A (en) * | 2013-09-04 | 2013-12-25 | 福建省农业科学院食用菌研究所 | Application of hypsizigus marmoreus lectin and deproteinization polysaccharide in preservation of loquats |
Non-Patent Citations (4)
| Title |
|---|
| PATRICK H.K. NGAI: "Lentin, a novel and potent antifungal protein from shitake mushroom with inhibitory effects on activity of human immunodeficiency virus-1 reverse transcriptase and proliferation of leukemia cells", 《LIFE SCIENCE》 * |
| 卢芳国: "《免疫学基础与病原生物学实验教程》", 31 August 2013 * |
| 吴丽萍: "4种食用菌提取物对2种植物病原真菌的抑制作用", 《安徽农业科学》 * |
| 麻兵继: "《药食兼用真菌重要次生代谢产物及其生物活性研究》", 30 November 2011 * |
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