CN100348109C - Method for extractiong microbicide and anti-staling agent from tupistra ker, and its application - Google Patents
Method for extractiong microbicide and anti-staling agent from tupistra ker, and its application Download PDFInfo
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- CN100348109C CN100348109C CNB2004100154754A CN200410015475A CN100348109C CN 100348109 C CN100348109 C CN 100348109C CN B2004100154754 A CNB2004100154754 A CN B2004100154754A CN 200410015475 A CN200410015475 A CN 200410015475A CN 100348109 C CN100348109 C CN 100348109C
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Abstract
The present invention discloses a method for extracting a microbicide or an anti-staling agent from tupistra ker and an application of a microbicide or an anti-staling agent. The root stocks or the herbs of plants of Tupistra Ker-Gawl are cut and pulverized to produce a pulverized object; the pulverized object is firstly extracted by water or an organic solvent, and an extracting solution is concentrated or dried to produce a water extract or an organic solvent extract; then the water extract or the organic solvent extract is orderly extracted by chloroform, ethyl acetate and normal butyl alcohol in fraction to produce a fraction extract through concentration or drying. The tupistra ker pulverized object, the water extract, the organic solvent extract or the fraction extract has the function of sterilizing and can be used as the microbicide or the anti-staling agent for fruits and vegetables. The tupistra ker pulverized object, the water extract, the organic solvent extract or the fraction extract of the present invention belongs to a novel plant source (type) pesticide, has obvious inhibiting effect on various crop pathogenic bacteria such as litchi downy mildew bacteria, banana anthracnose bacteria, cucumber downy mildew bacteria, paddy rice rhizoctonia solani, rice blast bacteria, etc. and can effectively prevent and cure litchi downy mildew, banana anthracnose and cucumber downy mildew.
Description
Technical field
The invention belongs to pesticide field, be specifically related to the preparation and the application of a class plant extracts, particularly the preparation of plant Tupistra chinensis Bak. extract and the application in control of plant disease thereof as bactericide and antistaling agent.
Background technology
Along with the further in-depth of the continuous development of human society and people's cognitive level, control pest and overcome its resistance, ensure food safety and protect environment more and more to be subjected to paying close attention to widely.At agriculture field, the abuse of chemical synthetic pesticide and the shortcoming of itself are to cause that damage by disease and insect produces resistance, the agricultural product residue of pesticide exceed standard and the principal element of environmental pollution.In order to address the above problem, people have strengthened the exploitation dynamics to biopesticide, and plant resource (property) agricultural chemicals therefore and is extremely paid attention to as the important component part of biopesticide, at present, the research and development report of plant resource (property) agricultural chemicals are arranged all both at home and abroad.The present invention adopts different extracting method to obtain the extract of this class plant of Tupistra chinensis Bak., prove through repetition test: the Tupistra chinensis Bak. extract has inhibitory action to many phytopathogens, especially peronophythora litchi, Glorosprium musarum Cookeet Mass, cucumber phytophthora, Rhizoctonia solani Kuhn and Pyricularia oryzae is had tangible fungistatic effect.The Tupistra chinensis Bak. extract can be used as plant resource (property) bactericide or fruit and vegetable fresh-keeping agent uses.
The Liliaceae Tupistra chinensis Bak. belongs to (Tupistra Ker-Gawl) plant, perennial herb.Root-like stock is slightly thick, and is near usually upright; Root is shorter, and the white wool of close life.Leaf usually base give birth to or consor on short stem, on the rare stem of being born in prolongation, narrow ellipse, lanceolar are to being with shape, gradually narrow one-tenth handle in bottom or handle are not obvious, stem is embraced in the base portion expansion.Calyx is by extracting out in the leafage, adnation, and upright or excurvation, base portion has first sheathing leaf; The flower that the spike tool is intensive; The full edge of bract becomes the tasselled shape; Perianth mitriform or cylindric, middle part or top 6 are split, and sepal is carried out; The goblet of the perianth rear portion inside expansion of tool sometimes; Stamen 6, because the perianth tube symphysis of filigree bottom, like the base portion of being born in perianth tube top or perianth sliver, flower pesticide is avette, carries, and is interior to lobe, filigree enlarges or does not enlarge; Ovary Room 3, every chamber 2-4 ovule, column cap expands, and what 3 split or are mushroom sometimes on the top.1-3 seed of berry tool.
Tupistra chinensis Bak. mainly is distributed in the Asia, and China has 12 kinds approximately, mainly produces each provinces and regions on the south the Changjiang river.
Summary of the invention
The present invention aims to provide plant Tupistra chinensis Bak. preparation method of extract and as the application of bactericide or fruit and vegetable fresh-keeping agent.To achieve these goals, the applicant is on the basis that many plants are extensively screened, found the sterilization idiocratic of Tupistra chinensis Bak., its extract not only shows higher stripped inhibition activity to the various plants pathogen, and under condition of living body, can prevent and treat peronophythora litchi, banana anthracnose and cucumber phytophthora root rot, fruits and vegetables harmless fresh-keeping had good effect.
The technical scheme that the present invention takes is: Tupistra chinensis Bak. is belonged to the root-like stock of (Tupistra Ker-Gawl) plant or herb section, segment, dry, dry in the shade or dry, pulverize, sieve crushed material, crushed material is soaked in water or organic solvent extraction, extract is through concentrating or dry water extract or extractive with organic solvent, water extract or extractive with organic solvent again with solvent carry out liquid-liquid or liquid-solid branch extract branch's extract, branch's extract is again through concentrating or dry branch's extract.Tupistra chinensis Bak. crushed material, water extract, extractive with organic solvent or branch's extract can be used as the usefulness of bactericide or antistaling agent.
Organic solvent of the present invention can be any one or a few of methyl alcohol, ethanol, acetone, benzene, dimethylbenzene, ethyl acetate etc., and the known material of this insider.
Above-mentioned Tupistra chinensis Bak. method for preparing extractive the steps include:
A. the root-like stock or the herb of the Tupistra chinensis Bak. of drying are pulverized, are sieved, crushed material.
B. get the Tupistra chinensis Bak. crushed material and place extraction element 3-8 times of water of volume ratio adding by weight, extract more than 24 hours, get aqueous extract.Or by weight volume ratio with 3-8 doubly to the extraction solvent of Tupistra chinensis Bak. crushed material weight, under refluxad extract, extracting temperature is 60～80 ℃, extraction time is 4～8 hours, or extracts about 24 hours, solvent extraction liquid.
C. Tupistra chinensis Bak. aqueous extract or organic solvent extraction liquid are concentrated or is concentrated into dried, concentrate or dry matter volume ratio by weight extract with 5-10 chloroform, ethyl acetate and n-butanol doubly branch successively again, concentrate then or dry branch's extract.
Prove repeatedly that through test Tupistra chinensis Bak. crushed material of the present invention, water extract, extractive with organic solvent or branch's extract have the obvious suppression effect to plant pathogenic fungis such as peronophythora litchi, Glorosprium musarum Cookeet Mass, cucumber phytophthora, Rhizoctonia solani Kuhn and Pyricularia oryzaes.And can effectively prevent and treat peronophythora litchi, banana anthracnose and cucumber phytophthora root rot.Concentration to control is: the concentration 〉=500mg/L of A. Tupistra chinensis Bak. water extract, extractive with organic solvent or branch's extract for preventing and treating peronophythora litchi; B. concentration 〉=the 400mg/L of Tupistra chinensis Bak. water extract, extractive with organic solvent or branch's extract for preventing and treating banana anthracnose; C. concentration 〉=the 300mg/L of Tupistra chinensis Bak. water extract, extractive with organic solvent or branch's extract for preventing and treating cucumber phytophthora root rot; D. concentration 〉=the 150mg/L of Tupistra chinensis Bak. water extract, extractive with organic solvent or branch's extract for preventing and treating rice sheath blight disease; E. concentration 〉=the 350mg/L of Tupistra chinensis Bak. water extract, extractive with organic solvent or branch's extract for preventing and treating rice blast.
Tupistra chinensis Bak. extract of the present invention is compared with existing common fungicide, and following characteristics are arranged:
1. the active ingredient in the environmentally safe Tupistra chinensis Bak. bactericide belongs to the natural antibacterial active substance, is the Secondary Metabolism of Plant product, can participate in the material and the energy metabolism of nature, imposes on the plant or in the environment, can not cause biological concentration, also unlikely cause residual.
2. comparatively safe to non-target organism.
3. the mechanism of action to phytopathogen is different from conventional pesticide, uses the back pathogen to be difficult for it is developed immunity to drugs.
4. have broad spectrum antibiotic activity, the various plants disease fungus is had inhibitory action.
Four, embodiment
(1) preparation embodiment
For a more clear understanding of the present invention and the drug effect of this extract, below the following embodiment that finishes according to technical solution of the present invention that provides by the inventor, the present invention is described in further detail, and these only are the present invention embodiment preferably, but be not limited to these embodiment.
The preparation of embodiment one Tupistra chinensis Bak. water extract
The Tupistra chinensis Bak. root-like stock or the herb of drying are pulverized, crossed 40～60 mesh sieves.In the Tupistra chinensis Bak. crushed material: the ratio of water=1: 3～8 (weight per volume), extract more than 24 hours, filter a part of extract, the residue centrifuge dripping gets another part extract and filters, merge two parts extract, again through vacuum concentration to heavy concentrate such as Tupistra chinensis Bak. crushed material or dry solid extract, low tempertaure storage is standby.
The preparation of embodiment two Tupistra chinensis Bak. extractive with organic solvent
The Tupistra chinensis Bak. root-like stock or the herb of drying are pulverized, crossed 40～60 mesh sieves.In the Tupistra chinensis Bak. crushed material: the ratio of methyl alcohol=1: 3～5 (weight per volume) places the heating of refluxing extraction device to extract, filter a part of extract, the residue centrifuge dripping gets another part extract and filters, merge two parts extract, again through vacuum concentration must with heavy concentrate such as Tupistra chinensis Bak. crushed material or dry solid extract, low tempertaure storage is standby.
The preparation of embodiment three Tupistra chinensis Bak. solvent branch extracts
With Tupistra chinensis Bak. water extract or extractive with organic solvent, place liquid distributing device, add the doubly chloroform of (weight per volume) of 5-10, leave standstill after fully stirring, treat mixed liquid clarification layering after, take out chloroform layer, vacuum concentration, chloroform extract, low tempertaure storage is standby; Remainder behind the chloroform extraction adds the doubly ethyl acetate of (weight per volume) of 5-10, leaves standstill after fully stirring, treat mixed liquid clarification layering after, take out ethyl acetate layer, vacuum concentration, ethyl acetate extract, low tempertaure storage is standby; Remainder after the ethyl acetate extraction adds the doubly n-butanol of (weight per volume) of 5-10, leaves standstill after fully stirring, treat mixed liquid clarification layering after, take out n-butanol layer, vacuum concentration, n-butanol extract, low tempertaure storage is standby.
(2) experimental example
Embodiment one Tupistra chinensis Bak. extract is to the inhibition activity test of plant pathogenic fungi
Under isolated condition, measured methanolic extract, ethyl acetate extract and the n-butanol extract of Tupistra chinensis Bak. inhibition activity with the mycelial growth rate method to the part plant pathogenic fungi, research report is as follows:
1, for examination bacterial classification peronophythora litchi (Peronophythora litchi), Glorosprium musarum Cookeet Mass (Collectotrichum musae), cucumber phytophthora (Phytophthora melonis), Rhizoctonia solani Kuhn (Rhizoctonia solani) and Pyricularia oryzae (Pyricularia oryzae).Bacterial classification is provided by gardening institute of Agricultural University Of South China, money ring institute and Institute of Plant Protection, academy of agricultural sciences, Guangdong Province.
2, test method mycelial growth rate method, the bacteriostasis rate computing formula is as follows:
The clean diameter of mycelial growth inhibition rate %=(the clean diameter of the contrast bacterium colony-clean diameter of processing bacterium colony) ÷ contrast bacterium colony * 100
3, result of the test Tupistra chinensis Bak. extract the results are shown in Table 1-3 to the virulence of plant pathogenic fungi mycelial growth:
Table 1 Tupistra chinensis Bak. methanolic extract is to the virulence of plant pathogenic fungi mycelial growth
| For the examination bacterial classification | The LD-P line | R | EC 50(mg/L) | EC 50Standard error | EC 5095% confidence limit (mg/L) |
| Peronophythora litchi | Y＝-8.36+4.17x | 0.99 | 1598.10 | 92.64 | 1426.45-1790.40 |
| Glorosprium musarum Cookeet Mass | Y＝10.69+5.06x | 0.94 | 1262.16 | 49.84 | 11168.15-1363.72 |
| Cucumber phytophthora root rot bacterium | Y＝0.11+1.44x | 0.99 | 2465.79 | 385.58 | 1814.88-3350.14 |
| Rhizoctonia solani Kuhn | Y＝-8.63+4.96x | 0.99 | 558.15 | 28.59 | 504.83-617.09 |
| Rice blast fungus | Y＝-6.04+3.51x | 0.99 | 1389.80 | 93.48 | 1218.14-1585.65 |
Table 2 Tupistra chinensis Bak. ethyl acetate extract is to the virulence of plant pathogenic fungi mycelial growth
| For the examination bacterial classification | The LLDD-PP line | R | EC 50(mg/L) | EC 50Standard error | EC 5095% confidence limit (mg/L) |
| Peronophythora litchi | Y＝-1.84+2.42X | 0.97 | 662.86 | 67.88 | 542.32-810.19 |
| Glorosprium musarum Cookeet Mass | Y＝-0.56+2.09X | 0.94 | 451.02 | 37.64 | 382.96-513.19 |
| Cucumber phytophthora root rot bacterium | Y＝0.25+1.88X | 0.89 | 337.53 | 32.34 | 279.75-4.7.25 |
| Rhizoctonia solani Kuhn | Y＝-3.34+3.75X | 0.98 | 167.80 | 27.28 | 112.01-230.78 |
| Rice blast fungus | Y＝0.16+1.88X | 0.98 | 369.90 | 50.69 | 282.77-483.89 |
Table 3 Tupistra chinensis Bak. n-butanol extract is to the virulence of plant pathogenic fungi mycelial growth
| For the examination bacterial classification | The LD-P line | R | EC 50(mg/L) | EC 50Standard error | EC 5095% confidence limit (mg/L) |
| Peronophythora litchi | Y＝-9.7+4.81x | 0.99 | 147.31 | 58.33 | 1038.48-1267.53 |
| Glorosprium musarum Cookeet Mass | Y＝12.78+5.97x | 0.98 | 955.58 | 43.67 | 873.71-1045.12 |
| Cucumber phytophthora root rot bacterium | Y＝-1.40+2.21x | 0.95 | 788.96 | 61.66 | 676.90-919.57 |
| Rhizoctonia solani Kuhn | Y＝-5.25+3.14x | 0.98 | 1847.29 | 101.20 | 1659.22-2056.68 |
| Rice blast fungus | Y＝-7.81+4.12x | 0.99 | 1288.12 | 76.14 | 1147.21-1446.34 |
Embodiment two Tupistra chinensis Bak. extracts are to the control test of peronophythora litchi
Selected lichee is washed fruit with surface disinfectant earlier, then wash, dry with running water, be that methanolic extract solution, ethyl acetate extract solution and the n-butanol extract solution of 1000mg/L soaks fruit 2min with concentration respectively then, dry, use white phytophthora sporangia suspension (2 * 10 again
5Individual/mL), and even spraying inoculation, the single fruit packing, after cultivating 4 days under 25 ℃ and the 98%RH condition, statistics disease index and control efficiency.Blank is established in test, and to prevent and treat the mould many Mils of bactericide thunder (Ridomil) of white epidemic disease as the contrast of commodity bactericide, many Mils of thunder solution concentration is 800mg/L.Result of the test such as table 4:
Table 4 Tupistra chinensis Bak. extract is to the control test of peronophythora litchi
| Experimental cultivar | Blank | The many Mil's contrasts of thunder | Methanolic extract | Ethyl acetate extract | N-butanol extract | ||||
| Disease refers to (%) | Disease refers to (%) | Preventive effect (%) | Disease refers to (%) | Preventive effect (%) | Disease refers to (%) | Preventive effect (%) | Disease refers to (%) | Preventive effect (%) | |
| The glutinous rice wrapped in lotus leaves imperial concubine laughs at the Huaihe River branch | 76.0 88.7 75.6 | 52.8 44.3 50.2 | 40.8 41.7 33.6 | 41.0 55.0 55.2 | 46.1 38.0 27.0 | 40.0 49.3 51.9 | 47.4 44.4 31.4 | 35.2 58.1 47.1 | 53.8 34.5 37.6 |
Embodiment three Tupistra chinensis Bak. extracts are to the control test of banana anthracnose
Selected banana (kind is " No. two, Guangdong ") is washed fruit with surface disinfectant earlier, then wash, dry with running water, soak fruit 2min with Tupistra chinensis Bak. water extract solution, methanolic extract solution, ethyl acetate extract solution and n-butanol extract solution respectively then, dry, after cultivating 15 days under 22-24 ℃ and the 98%RH condition, statistics disease index and control efficiency.Blank is established in test, and contrasts as the commodity bactericide with the bactericide sporgon (Sporgon) of preventing and treating anthracnose.Result of the test such as table 5:
Table 5 Tupistra chinensis Bak. extract is to the control efficiency of banana anthracnose
| Handle | Concentration (mg/L) | Disease refers to (%) | Preventive effect (%) |
| The positive alcohol extract of contrast sporgon water extract methanolic extract ethyl acetate extract | 0 1000 10000 1500 1000 1500 | 76.0 38.7 41.3 49.3 35.3 38.0 | 0 49.1 45.6 35.1 53.5 50.0 |
Embodiment four Tupistra chinensis Bak. extracts are to the control test of cucumber phytophthora root rot
Selected cucumber (kind is " a Japanese little green cucumber ") is washed fruit with surface disinfectant earlier, then wash, dry with running water, soak fruit 2min with Tupistra chinensis Bak. water extract solution, methanolic extract solution, ethyl acetate extract solution and n-butanol extract solution respectively then, dry, use cucumber phytophthora silk suspension (20/visual field) again, even spraying inoculation, single fruit packing, after cultivating 10 days under 25 ℃ and the 98%RH condition, statistics disease index and control efficiency.Blank is established in test, and contrasts as the commodity bactericide with the many Mils of bactericide thunder (Ridomil) of control phytophthora root rot.Result of the test such as table 6:
Table 6 Tupistra chinensis Bak. extract is to the control efficiency of cucumber phytophthora root rot
| Handle | Concentration (mg/L) | Disease refers to (%) | Preventive effect (%) |
| The positive alcohol extract of contrast thunder many Mils water extract methanolic extract ethyl acetate extract | 0 800 10000 2000 1000 2000 | 87.3 22.7 30.6 23.2 24.7 32.0 | 0 74.1 64.9 73.4 71.8 63.4 |
Claims (1)
1. Tupistra chinensis Bak. water extract, extractive with organic solvent or branch's extract is characterized in that as the purposes of bactericide: described Tupistra chinensis Bak. water extract, extractive with organic solvent or branch's extract are made by following steps:
A. the root-like stock or the herb of the Tupistra chinensis Bak. of drying are pulverized, are sieved, the Tupistra chinensis Bak. crushed material;
B. get the Tupistra chinensis Bak. crushed material, place the extraction element heavy water of 3-8 times of material of volume ratio adding by weight, extract more than 24 hours, get aqueous extract, concentrated then or drying obtains water extract; Or volume ratio adds the heavy 3-8 of material organic solvent doubly and places extraction element by weight, under refluxad extracts, and extracting temperature is 60～80 ℃, and extraction time is 4～8 hours, or lixiviate 24 hours, concentrates then or dry extractive with organic solvent;
C. with Tupistra chinensis Bak. water extract or extractive with organic solvent by weight volume ratio extract with 3-10 chloroform, ethyl acetate and n-butanol doubly branch successively, concentrate then or dry branch's extract;
The above organic solvent that extracts usefulness is any one or a few mixing such as methyl alcohol, ethanol, acetone, benzene, dimethylbenzene and ethyl acetate;
Described purposes is the control that is used for peronophythora litchi, banana anthracnose, cucumber phytophthora root rot, rice sheath blight disease and rice blast, and using method is spraying, immersion or coating; Wherein:
A. concentration 〉=the 500mg/L of Tupistra chinensis Bak. water extract, extractive with organic solvent or branch's extract for preventing and treating peronophythora litchi;
B. concentration 〉=the 400mg/L of Tupistra chinensis Bak. water extract, extractive with organic solvent or branch's extract for preventing and treating banana anthracnose;
C. concentration 〉=the 300mg/L of Tupistra chinensis Bak. water extract, extractive with organic solvent or branch's extract for preventing and treating cucumber phytophthora root rot;
D. concentration 〉=the 150mg/L of Tupistra chinensis Bak. water extract, extractive with organic solvent or branch's extract for preventing and treating rice sheath blight disease;
E. concentration 〉=the 350mg/L of Tupistra chinensis Bak. water extract, extractive with organic solvent or branch's extract for preventing and treating rice blast.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN100528205C (en) * | 2005-04-08 | 2009-08-19 | 三峡大学 | Tupistra Chinensis Bak extract medicinal composition, and its preparation method and use same |
| CN101401591B (en) * | 2008-11-24 | 2011-06-15 | 王占斌 | Method for extracting bactericide from wintergreen |
| CN105474943B (en) * | 2015-11-23 | 2018-07-31 | 湖北神农架国家级自然保护区管理局 | Tupistra cuttage breeding method |
| CN107793221A (en) * | 2017-11-01 | 2018-03-13 | 融安县飞腾中草药专业合作社 | A kind of coltsfoot solid fertilizer and preparation method thereof |
| CN108576201A (en) * | 2018-04-28 | 2018-09-28 | 广西红心食品有限公司 | A kind of antistaling agent for litchi and preparation method thereof |
| CN108576197A (en) * | 2018-04-28 | 2018-09-28 | 广西红心食品有限公司 | A kind of lichee fresh-keeping process |
| CN109758519B (en) * | 2019-03-07 | 2021-09-24 | 中国林业科学研究院林产化学工业研究所 | Preparation method and application of Tupistra chinensis extract |
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