CN103392746A - Application of Polygonum multiflorum extract - Google Patents
Application of Polygonum multiflorum extract Download PDFInfo
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- CN103392746A CN103392746A CN2013103748768A CN201310374876A CN103392746A CN 103392746 A CN103392746 A CN 103392746A CN 2013103748768 A CN2013103748768 A CN 2013103748768A CN 201310374876 A CN201310374876 A CN 201310374876A CN 103392746 A CN103392746 A CN 103392746A
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Abstract
The invention relates to an application of a Polygonum multiflorum extract in controlling grey mould fruit rot of strawberry. The extract is Polygonum multiflorum earthnut which is extracted by sterile water. According to the invention, Polygonum multiflorum is taken as a plant source extract resource to extract antibacterial substances, and the extract has the inhibiting effect and a certain in-vitro control efficiency to mould fruit rot of strawberry and can be further developed to novel plant source bactericide.
Description
Technical field
The invention belongs to biological pesticide technical field, relate generally to a kind of application of radix-polygoni multiflori extract, relate in particular to the application of radix-polygoni multiflori extract in controlling grey mould fruit rot of strawberry.
Background technology
The grey mould fruit rot of strawberry that is caused by Botrytis cinerea (Botrytis cinerea Pers.et Tris.) is one of Major Diseases of strawberry; the fruit of this disease main harm strawberry, the leaf of also can causing harm, cause Severe Reduction; even total crop failure, usually constitute a threat to the safety in production of protection ground strawberry.Since this disease, report is all arranged all over the world, become the restrictive barriers of strawberry volume increase.At present, in production, not yet find the anti-source of grey mould fruit rot of strawberry, the main chemical agent that adopts in control, wherein most importantly adopt benzimidazole and dicarboximide fungicide control, also there is report to use the medicaments such as procymidone, iprodione, phonetic mould amine, carbendazim, kresoxim-methyl, fludioxonil, but use continuously, make germ produce pesticide resistance, the preventive effect reduced year by year, also pollute environment.Along with the raising of people's quality of life, increasing to the demand of green safety noresidue strawberry, to be badly in need of seeking efficient, low toxicity, biogenic pesticide free from environmental pollution is prevented and treated Diseases of Strawberry.
Plant is the treasure-house of natural products agricultural chemicals, from plant, finding new bactericidal active substance, develops the novel plant source bactericide, is the focus of world today's novel pesticide exploitation.The fleece-flower root (Polygonum multiflo-rum Thunb.) claim again the tuber of multiflower knotweed, mountain essence, smart, the mountain tuber of multiflower knotweed, the red tuber of multiflower knotweed, redly hold back, the little only son and heir etc., belong to polygonaceae plant.It is reported, radix-polygoni multiflori extract mainly contains the pharmacologically active of following several respects: anti-oxidant, anti-ageing; Radiation proof, Green Tea Extract; Reducing blood lipid, anti arteriosclerosis; The effect of protection liver; Strengthen immunity of organisms; Improve memory, neuroprotective cell and intelligence development; The effects such as antibiotic, anti-inflammatory, analgesia.As can be seen here, the medical value of radix-polygoni multiflori extract is very extensive, and studies to obtain also comparison system, but this research that is extracted in control agricultural plant disease aspect has no report.In general, different with the object that plant is had to pathogenic microorganism effect owing to animal (people or beast) being had to the microorganism of pathogenic effects, pathogenic mode is also different, and the medicament that uses is inhibited to animal pathogen, but may not be inhibited to plant pathogen.Forefathers' result of study shows: radix-polygoni multiflori extract has certain inhibitory action to animal pathogen, but to the rarely seen report of the inhibiting research of phytopathogen.The present invention, according to this problem, has carried out Primary Study to the research of fleece-flower root retrieval inhibition agricultural plant disease-grey mould fruit rot of strawberry, and being intended to provides theoretical foundation for the prevention and control grey mould fruit rot of strawberry.
Summary of the invention
Technical problem to be solved by this invention is: for above-mentioned the deficiencies in the prior art, provide the control action of a kind of radix-polygoni multiflori extract to phytopathogen.
In order to solve the problems of the technologies described above, the technical solution adopted in the present invention is: the application of a kind of radix-polygoni multiflori extract in controlling grey mould fruit rot of strawberry.
Above-mentioned radix-polygoni multiflori extract is, after getting the piece root solarization dry grinding of the fleece-flower root, with sterile water, to carry out lixiviate, and gained filtrate evaporation and concentration again obtains to paste, is specially:
A, get after polygonum multiforum root tuber is cleaned dry to water content be 1.5-4.5%, through high speed disintegrator, with the rotating speed of 3000-5000r/min, be crushed to 75-95 order fine powder.
B, with sterile water, fleece-flower root powder is carried out to lixiviate, fleece-flower root powder is disposable to add, aseptic moisture adds for 3 times, after the ratio that at every turn adds the 30-50mL sterile water in 1g fleece-flower root powder adds, in on shaking table under 25-29 ℃ with 100-130r/min jolting 36-60h, filter, collect filtrate, merge 3 times filtrate;
C, gained filtrate being inserted to rotation concentration and evaporation instrument, is that 45-55 ℃ and condensation temperature are that under 5-9 ℃ of condition, 110-130r/min is concentrated into paste at 0.08-0.1MPa air pressure, bath temperature.
The above-mentioned radix-polygoni multiflori extract of carrying can adopt prior art to be prepared, and the present invention just adopts a wherein preparation method that the feasibility that it prepares is described.
Radix-polygoni multiflori extract of the present invention is by suppressing the modes such as conidia germination, Microsclerotia sprouting, inhibited to the Botrytis cinerea germ, simultaneously, this radix-polygoni multiflori extract has certain effect to preventing and treating stripped From Strawberry Leaves gray mold and stripped strawberry fruit gray mold, can be developed further into new botanical fungicide.And the extracting method of extract of the present invention is simple, quick, easy to operate, easily promotion and application; The required material of separating method is easily bought, and separating rate is fast, and is free from environmental pollution, to person poultry safety's environmental protection, saves cost, economizes on resources.
The accompanying drawing explanation
Fig. 1 is the stripped control efficiency figure of radix-polygoni multiflori extract of the present invention to the sick leaf of grey mould fruit rot of strawberry;
Wherein: 1 represents radix-polygoni multiflori extract; 2 representative contrast medicament carbendazim; 3 represent the blank sterile water.
Fig. 2 is the stripped control efficiency figure of radix-polygoni multiflori extract of the present invention to the grey mould fruit rot of strawberry fruit;
Wherein: 1 represents radix-polygoni multiflori extract; 2 representative contrast medicament carbendazim; 3 represent the blank sterile water.
Embodiment
Below in conjunction with concrete test and embodiment, technical scheme of the present invention is described further.
Raw material polygonum multiforum root tuber used in the present invention can or be bought the production technology maturation by plantation; The raw material supply abundance, the acquisition approach is various.
The pathogen species Botrytis cinerea germ of using in embodiment (Botrytis cinerea Pers.et Tris.) is provided by Agricultural University Of Hunan's plant pathology laboratory.The activation of this bacterial classification: adopt oese scraping 3 ring Botrytis cinerea germ bacterial classifications access potato dextrose agar slant mediums, cultivate 24-48h under 28 ℃.
Get after polygonum multiforum root tuber is cleaned and dry, with high speed disintegrator, with the rotating speed of 3000r/min, be crushed to 75 order fine powders, acquisition fleece-flower root powder.Getting fleece-flower root powder 10g packs in triangular flask, with sterile water, fleece-flower root powder is carried out to lixiviate, aseptic moisture adds for 3 times, after adding 30mL at every turn, put triangular flask on shaking table under 25 ℃ with 100r/min jolting 60h, filter, collect filtrate, merge 3 times filtrate, filtrate is packed into and rotated the concentration and evaporation instrument, at 0.08MPa air pressure, bath temperature, be that 45 ℃ and condensation temperature are under 5 ℃ of conditions, the 110r/min evaporation and concentration, to paste, obtains the reddish yellow radix-polygoni multiflori extract.
Get after the fleece-flower root is cleaned and dry, with high speed disintegrator, with the rotating speed of 4000r/min, be crushed to 85 order fine powders, acquisition fleece-flower root powder.Getting fleece-flower root powder 10g packs in triangular flask, with sterile water, fleece-flower root powder is carried out to lixiviate, aseptic moisture adds for 3 times, after adding 40mL at every turn, put triangular flask on shaking table under 27 ℃ with 115r/min jolting 48h, filter, collect filtrate, merge 3 times filtrate, filtrate is inserted to rotation concentration and evaporation instrument, at 0.09MPa air pressure, bath temperature, be that 50 ℃ and condensation temperature are under 7 ℃ of conditions, the 120r/min evaporation and concentration, to paste, obtains the reddish yellow radix-polygoni multiflori extract.
Get after the fleece-flower root is cleaned and dry, with high speed disintegrator, with the rotating speed of 5000r/min, be crushed to 95 order fine powders, acquisition fleece-flower root powder.Getting fleece-flower root powder 10g packs in triangular flask, with sterile water, fleece-flower root powder is carried out to lixiviate, aseptic moisture adds for 3 times, after adding 50mL at every turn, put triangular flask on shaking table under 29 ℃ with 130r/min jolting 36h, filter, collect filtrate, merge 3 times filtrate, filtrate is inserted to rotation concentration and evaporation instrument, at 0.1MPa air pressure, bath temperature, be that 55 ℃ and condensation temperature are under 9 ℃ of conditions, the 130r/min evaporation and concentration, to paste, obtains the reddish yellow radix-polygoni multiflori extract.
The inhibition of example 4 radix-polygoni multiflori extracts to the Botrytis cinerea germ
The radix-polygoni multiflori extract that takes above-mentioned preparation is packed in triangular flask, and with sterile water, being diluted to concentration is 35mg/mL solution, then takes 1mL solution and mix with 9mL potato dextrose agar (adding 40 μ g/mL streptomycins), shakes up and makes flat board.The inoculation diameter is the grey mould fruit rot of strawberry germ bacterium cake horizontalization plate central authorities of 5mm, with carbendazim (98% carbendazim active compound is with obtaining after the 0.1mol/L diluted hydrochloric acid dissolution), do the medicament contrast simultaneously, sterile water is made blank, every processing repeats 3 times, under 28-30 ℃, cultivate, after 48h, measure colony diameter, calculate inhibiting rate, the results are shown in following table 1.
Bacteriostasis rate (%)=[(control group colony diameter-processed group colony diameter)/(control group colony diameter-5)] * 100%.
The inhibitory action of table 1 radix-polygoni multiflori extract of the present invention to the Botrytis cinerea germ
Annotate: different lowercases represent significant difference (p<0.05), and different capitalizations represent utmost point significant difference (p<0.01).
Results and analysis: radix-polygoni multiflori extract of the present invention has certain inhibition to the Botrytis cinerea germ.The indoor antibacterial test result shows: after cultivating 48h, radix-polygoni multiflori extract of the present invention is 70.26% to Botrytis cinerea germ inhibiting rate, a little less than the inhibition of contrast medicament carbendazim.The results of analysis of variance shows: radix-polygoni multiflori extract is processed and contrast medicament carbendazim is processed significant difference, illustrates that radix-polygoni multiflori extract can, be used to preventing and treating the Botrytis cinerea germ, have the potentiality of further exploitation botanical fungicide.
The impact of example 5 radix-polygoni multiflori extract of the present invention on the spore germination of grey mould fruit rot of strawberry bacterium molecule
Adopt slide sprouting method to detect the impact of radix-polygoni multiflori extract on Botrytis cinerea germ conidia germination.at first by radix-polygoni multiflori extract, with the sterile water dilution, be the solution of 35mg/mL, simultaneously, Botrytis cinerea germ bacterium cake is placed on the potato dextrose agar flat board and cultivates 96h, after conidium grows, with the sterile water dilution, it is the spore suspension of 1 * 106/mL, get the conidial suspension of equivalent and mix rear dripping on slide with the solution after aforementioned dilution, take carbendazim as the contrast medicament, and establish blank, repeat 3 times, 48h is cultivated in moisturizing in culture dish, examine under a microscope the conidia germination situation, calculate spore germination rate, the results are shown in following table 2.
The impact of table 2 radix-polygoni multiflori extract on spore germination
Annotate: different lowercases represent significant difference (p<0.05), and different capitalizations represent utmost point significant difference (p<0.01).
Results and analysis: radix-polygoni multiflori extract and carbendazim all can suppress conidial sprouting consumingly, and wherein radix-polygoni multiflori extract is processed Botrytis cinerea germ sclerotium germination rate lower than without radix-polygoni multiflori extract, processing, but processes higher than contrast medicament carbendazim.The results of analysis of variance shows: radix-polygoni multiflori extract is processed and contrast medicament carbendazim is processed significant difference.Due to conidium, can cause protection ground grey mould fruit rot of strawberry and infect again, cause the underproduction, therefore, by using radix-polygoni multiflori extract, suppress conidia germination, thereby reduce source of infection again, alleviate disease and occur.
The impact that example 6 radix-polygoni multiflori extracts are sprouted Botrytis cinerea germ sclerotium
The Botrytis cinerea germ is inoculated on the potato dextrose agar flat board, cultivates 7d for 20 ℃, when the sclerotium blackening is ripe, collect sclerotium stand-by.Sclerotium is immersed in to 48h in the solution that contains series concentration (0.1mg/mL, 10mg/mL, 20mg/mL, 40mg/mL, 80mg/mL) radix-polygoni multiflori extract, air-dry being placed on the PDA flat board; To with the sclerotium that radix-polygoni multiflori extract is processed, not be placed on the PDA culture medium flat plate that contains above-mentioned series concentration radix-polygoni multiflori extract, 20 ℃ of cultivations, every processing repeats 3 times, 100 sclerotium of each replication.Relatively the sclerotium of two kinds of processing is sprouted situation, measures the sclerotium germination rate, the results are shown in following table 3.
Results and analysis: as shown in Table 3, through radix-polygoni multiflori extract, process the sclerotium germination rate and be starkly lower than the germination rate of processing without radix-polygoni multiflori extract, and along with the rising of radix-polygoni multiflori extract concentration, the sclerotium germination rate reduces gradually, wherein when drug concentration is brought up to 80mg/mL, the germination rate of sclerotium is minimum, illustrates that radix-polygoni multiflori extract sprouts and to play inhibitory action Microsclerotia.
The results of analysis of variance shows: the sclerotium germination rate significant difference that different radix-polygoni multiflori extracts are processed, but it is also remarkable to contrast sclerotium difference, and this may be not quite identical relevant with the sclerotium size.Because sclerotium can be resisted unsuitable environmental condition, can help pathogen to cross poor environment, the present invention finds that after deliberation radix-polygoni multiflori extract can reduce the sclerotium germination rate, reduces Botrytis cinerea germ primary source of infection, thereby reduces the generation of disease.As can be seen here, radix-polygoni multiflori extract not only suppresses the sprouting of sclerotium, and suppresses mycelial growth.
The impact that table 3 fleece-flower root is sprouted Botrytis cinerea germ Microsclerotia
| Concentration (mg/mL) | Process sclerotium germination rate (%) | Untreated sclerotium germination rate (%) |
| 0.1 | 82.35aA | 93.24cC |
| 10 | 74.71bB | 92.21dD |
| 20 | 73.83cC | 94.22bB |
| 40 | 61.15dD | 93.41cC |
| 80 | 62.32eE | 95.59aA |
Annotate: different lowercases represent significant difference (p<0.05), and different capitalizations represent utmost point significant difference (p<0.01).
The stripped control efficiency of example 7 radix-polygoni multiflori extracts to the From Strawberry Leaves gray mold
Win growth consistent, without the normal From Strawberry Leaves of damage by disease and insect, first use sterile water wash leaf table impurity, then with 75% alcohol, carry out surface sterilization 5s, then, with in sterile water, washing out residual alcohol, be placed in the vent cabinet natural air drying.Get respectively radix-polygoni multiflori extract, carbendazim medicament, sterile water and evenly be applied in the From Strawberry Leaves tow sides, be placed in the vent cabinet natural air drying, with the card punch of internal diameter 5mm, buy the fresh bacterium cake of getting the botrytis cinerea colony edge that has activated, with oese, choose to blade, during inoculation, avoid bacterium cake contact vein position, smear after complete blade is placed in culture dish, in temperature 26-28 ℃ of lower moisturizing, cultivate 3d, 3 blades of every processing also repeat 3 times, adopt the right-angled intersection method to measure strawberry fruit scab diameter, calculate control efficiency, the results are shown in following table 4.
Control efficiency (%)=[(contrast sick spot diameter – and process the scab diameter)/(contrast scab diameter-5)] * 100%.
Results and analysis: as shown in Figure 1, radix-polygoni multiflori extract on the From Strawberry Leaves eubolism without impact, and the From Strawberry Leaves gray mold scab diameter processed of radix-polygoni multiflori extract significantly less than blank, process after ash arrhizus bacteria infect the scab diameter of formation, but process greater than contrast medicament carbendazim.By measuring the scab diameter, calculate control efficiency, found that, the protection effect of radix-polygoni multiflori extract is 49.68%, a little less than the control efficiency of contrast medicament.Illustrate by constantly exploring the best diseases prevention concentration of radix-polygoni multiflori extract, time etc., it can be developed as to biological prevention and control agent.
The stripped control efficiency of table 4 radix-polygoni multiflori extract of the present invention to the From Strawberry Leaves gray mold
| Process | Colony diameter (mm) | Average colony diameter (mm) | Preventive effect (%) |
| Ⅰ | Ⅱ | Ⅲ | |||
| Radix-polygoni multiflori extract | 29.87 | 28.88 | 28.7 | 29.15 | 49.68bB |
| Carbendazim | 22.35 | 23.35 | 22.76 | 22.82 | 62.86aA |
| The sterile water contrast | 53.43 | 53.2 | 52.34 | 52.99 |
Annotate: different lowercases represent significant difference (p<0.05), and different capitalizations represent utmost point significant difference (p<0.01).
The stripped control efficiency of example 8 radix-polygoni multiflori extracts to the strawberry fruit gray mold
Win fruit size consistent, the strawberry fruit without the maturation of damage by disease and insect, first carry out surface sterilization 3s with 75% alcohol, then use the alcohol of sterile water wash residual, is placed in the vent cabinet natural air drying.Get radix-polygoni multiflori extract and smear with aseptic absorbent cotton at the malicious fruit surface of grass, be placed in the vent cabinet natural air drying.With sterile water, by activating on the botrytis cinerea bacterium colony of 72h the fresh botrytis cinerea spore of giving birth to, wash down, be mixed with 1 * 10
6The concentration of individual/mL, with watering can, at the strawberry fruit base portion, spray the circular spot of 5mm, with carbendazim medicament in contrast, and make blank with sterile water, fruit is placed on to moisturizing in the culture dish that diameter is 15cm and cultivates 48h, 3 fruits of every processing are also repeated 3 times, and utilize moccasin chi right-angled intersection method to measure colony diameter, calculate control efficiency, the results are shown in following table 5.
Control efficiency (%)=[(contrast colony diameter-processing colony diameter)/(contrast colony diameter-5)] * 100.
Results and analysis: as shown in Figure 2, radix-polygoni multiflori extract on the strawberry fruit eubolism without impact, and the scab diameter that ash arrhizus bacteria infects formation after radix-polygoni multiflori extract is processed significantly infects the scab diameter of formation less than the ash arrhizus bacteria of control treatment, but greater than the scab diameter of contrast medicament carbendazim.Simultaneously, by the right-angled intersection method, measure the scab diameter, calculate control efficiency and find: radix-polygoni multiflori extract to the control efficiency of grey mould fruit rot of strawberry fruit lower than contrast medicament carbendazim.The results of analysis of variance shows: radix-polygoni multiflori extract is processed and contrast medicament carbendazim is processed significant difference.Illustrate that radix-polygoni multiflori extract can be used for preventing and treating grey mould fruit rot of strawberry.
The stripped control efficiency of table 5 radix-polygoni multiflori extract of the present invention to the strawberry fruit gray mold
| Process | Colony diameter (mm) | Average colony diameter (mm) | Preventive effect (%) |
| Ⅰ | Ⅱ | Ⅲ | |||
| Radix-polygoni multiflori extract | 29.8 | 30.08 | 30.07 | 29.98 | 36.93bB |
| Carbendazim | 25.76 | 25.25 | 25.7 | 25.57 | 48.08aA |
| The sterile water contrast | 45.53 | 44.34 | 44.01 | 45 |
Annotate: different lowercases represent significant difference (p<0.05), and different capitalizations represent utmost point significant difference (p<0.01).
Claims (5)
1. the application of radix-polygoni multiflori extract in controlling grey mould fruit rot of strawberry.
2. the application of radix-polygoni multiflori extract in From Strawberry Leaves gray mold and stripped strawberry fruit gray mold are exsomatized in control.
3. application as claimed in claim 1 or 2 is characterized in that: described radix-polygoni multiflori extract is that the piece root extraction of getting the fleece-flower root forms.
4. application as claimed in claim 1 or 2 is characterized in that: described radix-polygoni multiflori extract is the water extract of the fleece-flower root.
5. application as claimed in claim 4 is characterized in that: described radix-polygoni multiflori extract is, after getting polygonum multiforum root tuber solarization dry grinding, with sterile water, to carry out lixiviate, and gained filtrate evaporation and concentration again obtains to paste.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104041538A (en) * | 2014-05-28 | 2014-09-17 | 赵兰 | Natural fungicide for prevention and control of strawberry black spot and preparation method thereof |
| CN105104457A (en) * | 2015-09-11 | 2015-12-02 | 湖州旭龙生物化学有限公司 | Bactericide composition |
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| CN1515152A (en) * | 2003-08-29 | 2004-07-28 | 湖北省农业科学植保土肥研究所 | Application of antraquinone derivative as pesticide for controlling plant diseases |
| CN101961326A (en) * | 2010-10-29 | 2011-02-02 | 西南交通大学 | Application of physcion in preparing medicines for treating depression |
| CN102318651A (en) * | 2011-07-28 | 2012-01-18 | 北京农学院 | Nematicidal extract of radix-polygoni multiflori and preparation method thereof |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1515152A (en) * | 2003-08-29 | 2004-07-28 | 湖北省农业科学植保土肥研究所 | Application of antraquinone derivative as pesticide for controlling plant diseases |
| CN101961326A (en) * | 2010-10-29 | 2011-02-02 | 西南交通大学 | Application of physcion in preparing medicines for treating depression |
| CN102318651A (en) * | 2011-07-28 | 2012-01-18 | 北京农学院 | Nematicidal extract of radix-polygoni multiflori and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104041538A (en) * | 2014-05-28 | 2014-09-17 | 赵兰 | Natural fungicide for prevention and control of strawberry black spot and preparation method thereof |
| CN104041538B (en) * | 2014-05-28 | 2015-11-18 | 赵兰 | A kind ofly prevent and treat natural bactericidal agent of strawberry black spot and preparation method thereof |
| CN105104457A (en) * | 2015-09-11 | 2015-12-02 | 湖州旭龙生物化学有限公司 | Bactericide composition |
| CN105104457B (en) * | 2015-09-11 | 2018-07-10 | 湖州旭龙生物化学有限公司 | A kind of microbicide compositions |
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