CN103222486A - Plant extract with inhibiting effect on orange pathogenic bacteria as well as preparation method and application thereof - Google Patents
Plant extract with inhibiting effect on orange pathogenic bacteria as well as preparation method and application thereof Download PDFInfo
- Publication number
- CN103222486A CN103222486A CN2013101953946A CN201310195394A CN103222486A CN 103222486 A CN103222486 A CN 103222486A CN 2013101953946 A CN2013101953946 A CN 2013101953946A CN 201310195394 A CN201310195394 A CN 201310195394A CN 103222486 A CN103222486 A CN 103222486A
- Authority
- CN
- China
- Prior art keywords
- extract
- walnut
- peel
- green
- penicillium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a preparation method for extracting an extract with an inhibiting effect on orange pathogenic bacteria from walnut green seedcases and an application of the extract in prevention and treatment of plant diseases caused by the pathogenic bacteria.
Description
Technical field
The present invention relates to a kind ofly has inhibiting plant extracts and preparation method thereof to the citrus pathogen.The invention belongs to plant extracts and antibacterial activity field thereof.
Background technology
Scutellum rot of orange and green mould are respectively the plant diseases that is caused by penicillium italicum (Penicillium italium) and green mold bacterium (Penicillium digitaum).According to statistics, the loss that citrus fruit causes because of decayed fruit in transporting procedures accounts for more than 90% based on scutellum rot of orange and green mould.At present, the prophylactico-therapeutic measures of the citrus disease of extensive use in the production is employing chemical bactericide such as probenazole, presses down mould azoles, sodium-o-phenyl phenolate (SOPP) etc. that these medicaments use existing historical for many years as the oranges and tangerines preservative.Yet, because people are to the unreasonable use and the intrinsic shortcoming of medicine itself of chemicals, cause chemical pesticide in use to produce problems such as environmental pollution, person poultry poisoning, residue of pesticide, along with the raising of people's quality of life and the enhancing of environmental consciousness, the public is more and more higher to the cry of restriction chemical bactericide.In order to seek to replace the new method of chemical bactericide, people have turned to safe, stable, pollution-free, natural drug efficiently to sight.
Monarch (1989) discovers just now, and geranium oil, citronella oil have the obvious suppression effect to the citrus Penicillium notatum, and inferior is spearmint oil, and the antibacterial power of fragrant Pu's oil is the most weak.Still the strongest with geranium oil and citronella oil to the bacteriostatic activity of citrus green mold for the examination essential oil, spearmint oil is much lower than the above two, and fragrant flower bud oil is then lower.(1993) such as Xi Yu virtues are discovered, the juice of garlic, onion, ginger all has inhibition and killing action to citrus green grass or young crops, green mold bacterium, and wherein garlic rhythm bacteriostasis performance is the strongest, and onion takes second place, and ginger is then relatively poor.Xu Qiyun etc. (2006) discover, ginkgo leaf ethanol extract, petroleum ether extract, chloroform extraction thing and 4 kinds of extracts of water extract all have certain inhibition effect to citrus pathogens penicillium, but have only petroleum ether extract to the citrus pathogens penicillium good antimicrobial effect.Qiu Qi etc. (2008) discover that the extract of tea seed, Alocasia ordora, mountain villous themeda orchid, water eggplant, Shanghai leaf Chinese ilex, mahogany, arbor-vitae, Michelia maudiae etc. has certain bacteriostatic activity to the oranges and tangerines penicillium.Hu Junhua etc. (2010) discover that the extract of Ligusticum wallichii, arbor-vitae, Siberian cocklebur and cortex acanthopanacis is stronger to the inhibitory action of citrus Penicillium notatum, and bacteriostasis rate is all more than 97%; The extract of cortex acanthopanacis and arbor-vitae is stronger to citrus green mold bacterium inhibitory action.Ceng Rong etc. (2010) discover, 11 plant species extracts show in various degree inhibition effect to Italian mould, and wherein the extract inhibitory action with plants such as CAULIS IMPATIENTIS, Pogostemon cablin, sweet wormwood, rhizoma calamis is comparatively obvious.In addition, Cao Tingting etc. (2008) have studied the inhibitory action of the prompt peace of bacillus subtilis (Bacillus subtilis) ZK strain fermentation product peptide element to the citrus penicillium, prove that prompt peace peptide element is plain more responsive to victory peace peptide than Penicillium notatum to citrus green mold bacterium on the PDA flat board.
The activity of anti-citrus penicillium has been studied in above-mentioned research from aspects such as plant extracts, microbial fermentation products, also found to have the extract of certain antibacterial activity, but the antibacterial effect of extract is unsatisfactory, and mostly used extraction material is economical or medicinal species.Can find and a kind ofly not only have good antibacterial activity but also more cheap material, and then obtain antibacterial substance and the extracting method that has more practical value?
Based on above background, the inventor is a raw material with common green peel of walnut as discarded object, has carried out to have the oranges and tangerines of inhibition green grass or young crops, the extracting method of green mold fungus extract and the research of bacteriostasis.It below is particular content of the present invention.
Summary of the invention
The object of the invention provides a kind ofly has inhibiting plant extracts and preparation method thereof to the citrus pathogen.
For achieving the above object, the present invention includes following technical scheme:
The present invention at first provides a kind of extracting method of walnut peel extract, this method comprises: place 50 ℃ of drying in oven to constant weight and pulverizing green peel of walnut, cross 90 mesh sieves, taking by weighing the green peel of walnut powder is to extract solvent with the absolute methanol, solid-liquid ratio is 1:15,65 ℃ of following refluxing extraction 3 times, each 2h merges the extract concentrating under reduced pressure and becomes the medicinal extract shape to constant weight.Then, with acetone-water (1:1) dissolving fully, use isopyknic benzinum, chloroform, ethyl acetate, extracting n-butyl alcohol successively, behind concentrating under reduced pressure, obtain each extraction phase extract of methanolic extract.
The present invention also provides the extracting method of another kind of walnut peel extract, this method comprises: green peel of walnut places 50 ℃ of drying in oven to constant weight and pulverizing, cross 90 mesh sieves, taking by weighing the green peel of walnut powder is to extract solvent with the benzinum, solid-liquid ratio is 1:15,65 ℃ of following refluxing extraction 3 times, each 2h merges the extract concentrating under reduced pressure and becomes the medicinal extract shape to constant weight.
Wherein, the solid-liquid ratio 1:15 that mentions in the said extracted method be mass/volume than (w/v), promptly 1 gram material is with 15 milliliters of solvents; The acetone-water of mentioning in the said method (1:1) for volume/volume than (v/v), promptly 1 milliliter of acetone mixes with 1 ml water.
Wherein, the gained ligroin extraction is the extract with antibacterial activity effect in the said extracted method.
Above-mentioned second method is directly concentrated by the benzinum extracting, has saved steps such as methyl alcohol extracting, solvent extraction, and is simple.
Secondly, the present invention also provides a kind of walnut peel extract, and this extract is the benzinum paste that obtains through concentrating under reduced pressure respectively according to the ligroin extraction that above-mentioned two kinds of extracting method obtain.
In addition, the present invention also provides above-mentioned walnut peel extract as the purposes in the antibacterial agent.
Preferably, the control of plant disease product of described purposes for above-mentioned walnut peel extract being used for as the active ingredient of antibacterial agent cause by penicillium italicum (Penicillium italium) and/or green mold bacterium (Penicillium digitaum).
Antibacterial activity of the present invention adopts inhibition zone determination method (Oxford agar diffusion method), and the positive control medicine is an amphotericin B.
Description of drawings
Fig. 1 is that the green peel of walnut ligroin extraction suppresses and can try hard to Penicillium notatum; Wherein Am B is a positive control, and A1-A5 is the fungistatic effect of concentration when 100-3.13mg/mL.
Embodiment
Explain the present invention below in conjunction with embodiment.Embodiment is for ease of better understanding the present invention, but is not limitation of the present invention.
Embodiment 1:
Green peel of walnut is in 50 ℃ of drying in oven to constant weight and pulverize, and crosses 90 mesh sieves, and taking by weighing the green peel of walnut powder is to extract solvent with the absolute methanol, and solid-liquid ratio is 1:15,65 ℃ of following refluxing extraction 3 times, and each 2h merges the extract concentrating under reduced pressure and becomes the medicinal extract shape to constant weight.Then, with acetone-water (1:1) dissolving fully, use isopyknic benzinum, chloroform, ethyl acetate, extracting n-butyl alcohol successively, behind concentrating under reduced pressure, obtain each extraction phase extract of methanolic extract.
Adopt Oxford agar diffusion method working sample bacteriostatic activity.Take by weighing a certain amount of different extract sample, be made into 50mg/mL with appropriate solvent, get the abundant mixing of medium that cultured bacterial suspension of 25 μ L and 20mL are cooled to 50 ℃, pour into and make the flat board that carries disease germs in the 9cm culture dish, the flat board that carries disease germs contains bacteria concentration about 10
6Individual/mL.With diameter is that the Oxford cup of 0.8cm lies on the flat board that carries disease germs, and adds testing sample 150 μ L, repeats 3 times.Place and cultivate the 72h(fungi in 25 ℃ of incubators), measure antibacterial circle diameter with the right-angled intersection method.With coordinative solvent as blank.
Experimental result: when being 50mg/mL for the test agent mass concentration, each extraction phase of green peel of walnut methanolic extract sees Table 1 and Fig. 1 to the inhibition effect of several harmful fungoids.
Each extraction phase of table 1 green peel of walnut methanolic extract suppresses ability
Wherein: no bacteriostatic activity (-), antibacterial circle diameter＜0.8cm; Weak bacteriostatic activity (+), antibacterial circle diameter 0.8-1.0cm; Medium bacteriostatic activity (++), antibacterial circle diameter 1.0～1.2cm; High bacteriostatic activity (+++), antibacterial circle diameter 1.2～1.4cm; Strong bacteriostatic activity (++ ++), antibacterial circle diameter 1.4～1.6cm; Strong bacteriostatic activity (++ +++), antibacterial circle diameter ﹥ 1.6cm; Standard deviation ± 0.5mm.
As can be seen from Table 1, green peel of walnut benzinum phase, chloroform all have certain bacteriostatic activity to penicillium italicum, citrus green mold bacterium when 50mg/mL, and wherein, the relative penicillium italicum of benzinum, citrus green mold bacterium fungistatic effect are best; Chloroform also has faint bacteriostatic activity mutually, but can not show a candle to the benzinum phase, and other extract and all do not detect bacteriostatic activity mutually.As seen, to citrus Penicillium notatum and green mold bacterium have Petroleum ether extraction that the composition that suppresses active mainly is distributed in green peel of walnut mutually in.
Simultaneously, as positive control, find that the bacteriostasis of 25mg/mL ligroin extraction is suitable with 200 μ g/mL amphotericin Bs with amphotericin B.
Embodiment 2:
Each preparation method of extract adopts the Oxford agar diffusion method to measure minimum inhibitory concentration with embodiment 1.Take by weighing a certain amount of each extract, be configured as 100mg/mL with coordinative solvent, doubling dilution becomes 100,50,25,12.5,6.25,3.13mg/mL.According to the test examples 1 arrangements of steps flat board that carries disease germs, draw 150 μ L respectively, repeat 3 times.Place and cultivate 72h in 25 ℃ of incubators, just do not have the minimum inhibitory concentration that is of inhibition zone.With coordinative solvent as blank.
Each extraction phase minimum inhibitory concentration of table 2 green peel of walnut methanolic extract
As can be seen from Table 2, the green peel of walnut ligroin extraction all has fungistatic effect preferably to penicillium italicum and citrus green mold bacterium, ligroin extraction to the bacteriostasis MIC of penicillium italicum, citrus green mold bacterium be respectively 3.125,6.25mg/mL.
Embodiment 3
Green peel of walnut places 50 ℃ of drying in oven to constant weight and pulverize, and crosses 90 mesh sieves, and taking by weighing the green peel of walnut powder directly is to extract solvent with the benzinum, and solid-liquid ratio is 1:15,65 ℃ of following refluxing extraction 3 times, and each 2h merges the extract concentrating under reduced pressure and becomes the medicinal extract shape to constant weight.This extract is carried out antibacterial activity measure (assay method is except that the doubling dilution initial concentration is 80mg/mL, with experiment embodiment 2), the result is as shown in table 3.
The bacteriostatic activity of table 3 green peel of walnut ligroin extraction
The above results shows, directly still to have and experiment embodiment 1 and 2 suitable effects with benzinum extracting, the concentrated extract that obtains even saved steps such as methyl alcohol extracting, solvent extraction, and is more simple through optimizing back extraction preparation method.
Claims (5)
1. the extracting method of a walnut peel extract, this method comprises: place 50 ℃ of drying in oven to constant weight and pulverizing green peel of walnut, cross 90 mesh sieves, taking by weighing the green peel of walnut powder is to extract solvent with the absolute methanol, solid-liquid ratio is 1:15,65 ℃ of following refluxing extraction 3 times, each 2h merges the extract concentrating under reduced pressure and becomes the medicinal extract shape to constant weight.Then, complete with the acetone-water dissolving of 1:1, use isopyknic benzinum, chloroform, ethyl acetate, extracting n-butyl alcohol successively, behind concentrating under reduced pressure, obtain each extraction phase extract of methanolic extract;
Wherein, the solid-liquid ratio 1:15 that mentions in the said extracted method be mass/volume than (w/v), promptly 1 gram material is with 15 milliliters of solvents; The acetone-water of the 1:1 that mentions in the said method be volume/volume than (v/v), promptly 1 milliliter of acetone mixes with 1 ml water.
2. the extracting method of walnut peel extract according to claim 1, this method comprises: green peel of walnut places 50 ℃ of drying in oven to constant weight and pulverizing, cross 90 mesh sieves, taking by weighing the green peel of walnut powder is to extract solvent with the benzinum, solid-liquid ratio is 1:15,65 ℃ of following refluxing extraction 3 times, each 2h merges the extract concentrating under reduced pressure and becomes the medicinal extract shape to constant weight;
Wherein, the solid-liquid ratio 1:15 that mentions in the said extracted method be mass/volume than (w/v), promptly 1 gram material is with 15 milliliters of solvents.
3. a walnut peel extract is characterized in that, described extract is the benzinum paste that obtains through concentrating under reduced pressure according to the ligroin extraction that the method for claim 1 or 2 obtains.
4. the described walnut peel extract of claim 3 is as the purposes in the antibacterial agent.
5. purposes according to claim 4, it is characterized in that the control of plant disease product of described purposes for above-mentioned walnut peel extract being used for as the active ingredient of antibacterial agent cause by penicillium italicum (Penicillium italium) and/or green mold bacterium (Penicillium digitaum).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310195394.6A CN103222486B (en) | 2013-05-23 | 2013-05-23 | Plant extract with inhibiting effect on orange pathogenic bacteria as well as preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310195394.6A CN103222486B (en) | 2013-05-23 | 2013-05-23 | Plant extract with inhibiting effect on orange pathogenic bacteria as well as preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103222486A true CN103222486A (en) | 2013-07-31 |
| CN103222486B CN103222486B (en) | 2014-11-19 |
Family
ID=48833320
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310195394.6A Expired - Fee Related CN103222486B (en) | 2013-05-23 | 2013-05-23 | Plant extract with inhibiting effect on orange pathogenic bacteria as well as preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103222486B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103461470A (en) * | 2013-09-03 | 2013-12-25 | 新疆农业科学院农产品贮藏加工研究所 | Walnut green seedcase extract compound preservative as well as preparation method and application thereof |
| CN104605013A (en) * | 2014-10-08 | 2015-05-13 | 江西农业大学 | Preparation method of thalictrum plant extract, thalictrum plant extract prepared according to preparation method, and application of thalictrum plant extract |
| CN107821447A (en) * | 2017-11-30 | 2018-03-23 | 孙铭岐 | A kind of walnut peel extract bactericide and its application process in preventing and treating Fusarium solani |
| CN109111300A (en) * | 2018-11-06 | 2019-01-01 | 山东省葡萄研究院 | A kind of grape special nutritional bacteriostatic agent and the preparation method and application thereof |
| CN110402970A (en) * | 2019-08-21 | 2019-11-05 | 贵州农鹰科技有限公司 | A kind of plant lemon is dispelled bacterium liquid and the preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101416644A (en) * | 2008-01-21 | 2009-04-29 | 北京农学院 | A kind of walnut peel extract and new purposes thereof |
| CN101906030A (en) * | 2010-08-23 | 2010-12-08 | 河北理工大学 | Method for extracting juglone from walnut green husk residue by microwave reflux |
| CN102579565A (en) * | 2012-01-18 | 2012-07-18 | 河北联合大学 | Method for microwave reflux extraction of general flavone in walnut peel residue |
-
2013
- 2013-05-23 CN CN201310195394.6A patent/CN103222486B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101416644A (en) * | 2008-01-21 | 2009-04-29 | 北京农学院 | A kind of walnut peel extract and new purposes thereof |
| CN101906030A (en) * | 2010-08-23 | 2010-12-08 | 河北理工大学 | Method for extracting juglone from walnut green husk residue by microwave reflux |
| CN102579565A (en) * | 2012-01-18 | 2012-07-18 | 河北联合大学 | Method for microwave reflux extraction of general flavone in walnut peel residue |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103461470A (en) * | 2013-09-03 | 2013-12-25 | 新疆农业科学院农产品贮藏加工研究所 | Walnut green seedcase extract compound preservative as well as preparation method and application thereof |
| CN104605013A (en) * | 2014-10-08 | 2015-05-13 | 江西农业大学 | Preparation method of thalictrum plant extract, thalictrum plant extract prepared according to preparation method, and application of thalictrum plant extract |
| CN104605013B (en) * | 2014-10-08 | 2017-12-01 | 江西农业大学 | A kind of preparation method of thalictrum spp extract and its thalictrum spp extract and purposes of preparation |
| CN107821447A (en) * | 2017-11-30 | 2018-03-23 | 孙铭岐 | A kind of walnut peel extract bactericide and its application process in preventing and treating Fusarium solani |
| CN109111300A (en) * | 2018-11-06 | 2019-01-01 | 山东省葡萄研究院 | A kind of grape special nutritional bacteriostatic agent and the preparation method and application thereof |
| CN110402970A (en) * | 2019-08-21 | 2019-11-05 | 贵州农鹰科技有限公司 | A kind of plant lemon is dispelled bacterium liquid and the preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103222486B (en) | 2014-11-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR101532005B1 (en) | Natural preservative comprising citrus junos seed extract and manufacturing method thereof | |
| CN103222486B (en) | Plant extract with inhibiting effect on orange pathogenic bacteria as well as preparation method and application thereof | |
| CN103238634B (en) | Application of amaranthus in plant disease prevention and treatment | |
| Obidi et al. | Antimicrobial activity of orange oil on selected pathogens | |
| KR101824687B1 (en) | Antimicrobial composition comprising natural plant extract | |
| KR101806800B1 (en) | Complex natural preservative comprising scutellaria baicalensis extract and method manufacture the same | |
| CN103913564A (en) | Method for testing bacteriostatic activity of spire wampee extractive | |
| CN101991715A (en) | Aquilaria sinensis fruit skin extract and preparation method and application thereof | |
| KR20160114206A (en) | A antimicrobial composition comprising scutellariae radix, schisandrae fructus, lonicerae flos, cnidi fructus and zanthoxyli fructus | |
| CN107602521A (en) | A kind of method and obtained OPC and purposes of pitaya peel extraction OPC | |
| CN105360171A (en) | Preparation method and application of plant source bacteriostatic agent | |
| CN109452320A (en) | The purposes of Fructus Caryophylli volatile oil | |
| CN105407725B (en) | The composition and method of selectivity control invasive species | |
| CN112772681B (en) | Biological compound disinfectant and preparation method thereof | |
| Rejeki et al. | Preliminary study of Hyptis pectinata (L.) poit extract biotransformation by Aspergillus niger | |
| CN102716188A (en) | Method for extracting extract from mangnolia officinalis fruit and antibacterial application thereof | |
| CN109221269A (en) | A kind of FLOS CHRYSANTHEMI ALBA from Haizhou of China leaf extract and preparation method thereof and the application in restraining and sterilizing bacteria | |
| CN101422419B (en) | Toilet water and manufacture method thereof | |
| CN102870824A (en) | Preparation method for active mixed extract of tea tree oil meal as well as extract and application of extract | |
| CN103004894A (en) | Biopesticide for preventing and treating powdery mildew of melons and fruits and preparation method of biopesticide | |
| CN109042651A (en) | The application of apple pomace and apple leaf extract in phytopathogen prevention and treatment | |
| CN106538624A (en) | A kind of coptis bactericide and its application in preventing and treating crop epidemic disease | |
| Abu-Salem et al. | Antimicrobial activity and phytochemicals screening of jojoba (Simmondsia chinensis) root extracts and latex | |
| KR102488821B1 (en) | Manufacturing method of tee-tree extract and antimicrobial compositon comprising the same | |
| CN109156478A (en) | A kind of Polyelement botanical preservative and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20141119 Termination date: 20210523 |