CN103392746B - Application of polygonum multiflorum extract - Google Patents
Application of polygonum multiflorum extract Download PDFInfo
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- CN103392746B CN103392746B CN201310374876.8A CN201310374876A CN103392746B CN 103392746 B CN103392746 B CN 103392746B CN 201310374876 A CN201310374876 A CN 201310374876A CN 103392746 B CN103392746 B CN 103392746B
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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 radix-polygoni multiflori extract in the application of controlling in grey mould fruit rot of strawberry.
Background technology
The grey mould fruit rot of strawberry being 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 constitutes a threat to the safety in production of protection ground strawberry.Since this disease, all there is report 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, preventive effect reduced year by year, also pollutes 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, finds new bactericidal active substance from plant, develops 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 antibacterial, 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 object plant to pathogenic microorganism effect owing to animal (people or beast) being there is to the microorganism of pathogenic effects, pathogenic mode is also different, the medicament using 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 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: a kind of radix-polygoni multiflori extract is at the application of controlling in grey mould fruit rot of strawberry, the water extract that this radix-polygoni multiflori extract is the fleece-flower root.
Above-mentioned radix-polygoni multiflori extract is that the piece root of getting the fleece-flower root shines after dry grinding, carries out lixiviate with sterile water, and gained filtrate again evaporation and concentration obtains to paste, is specially:
A, get polygonum multiforum root tuber clean after dry to water content be 1.5-4.5%, be crushed to 75-95 order fine powder through high speed disintegrator with the rotating speed of 3000-5000r/min.
B, fleece-flower root powder is carried out to lixiviate with sterile water, fleece-flower root powder is disposable to add, aseptic moisture adds for 3 times, after the ratio that at every turn adds 30-50mL sterile water in 1g fleece-flower root powder adds, on shaking table at 25-29 DEG C 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 DEG C and condensation temperature are that under 5-9 DEG C of condition, 110-130r/min is concentrated into paste at 0.08-0.1MPa air pressure, bath temperature.
Above-mentioned carried radix-polygoni multiflori extract can adopt prior art to be prepared, and the present invention just adopts a wherein preparation method that the feasibility that it is prepared is described.
Radix-polygoni multiflori extract of the present invention is by suppressing the modes such as conidia germination, Microsclerotia sprouting, inhibited to Botrytis cinerea germ, simultaneously, this radix-polygoni multiflori extract has certain effect to preventing and treating in vitro From Strawberry Leaves gray mold with in vitro 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, free from environmental pollution, to person poultry safety's environmental protection, saves cost, economizes on resources.
Brief description of the drawings
Fig. 1 is the in vitro 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 blank sterile water.
Fig. 2 is the in vitro control efficiency figure of radix-polygoni multiflori extract of the present invention to grey mould fruit rot of strawberry fruit;
Wherein: 1 represents radix-polygoni multiflori extract; 2 representative contrast medicament carbendazim; 3 represent 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 be by planting or buying, production technology maturation; Raw material supply abundance, acquisition approach is various.
The pathogen species Botrytis cinerea germ (Botrytis cinerea Pers.et Tris.) of using in embodiment is provided by Agricultural University Of Hunan's plant pathology laboratory.The activation of this bacterial classification: adopt oese scraping 3 to encircle Botrytis cinerea germ bacterial classification access potato dextrose agar slant medium, cultivate 24-48h at 28 DEG C.
Embodiment 1
Get after polygonum multiforum root tuber is cleaned and dry, be crushed to 75 order fine powders with the rotating speed of 3000r/min with high speed disintegrator, acquisition fleece-flower root powder.Getting fleece-flower root powder 10g packs in triangular flask, fleece-flower root powder is carried out to lixiviate with sterile water, aseptic moisture adds for 3 times, adds after 30mL at every turn, put triangular flask on shaking table at 25 DEG C with 100r/min jolting 60h, filter, collect filtrate, merge 3 times filtrate, pack filtrate into rotation concentration and evaporation instrument, be that 45 DEG C and condensation temperature are under 5 DEG C of conditions at 0.08MPa air pressure, bath temperature, 110r/min evaporation and concentration, to paste, obtains reddish yellow radix-polygoni multiflori extract.
Embodiment 2
Get after the fleece-flower root is cleaned and dry, be crushed to 85 order fine powders with the rotating speed of 4000r/min with high speed disintegrator, acquisition fleece-flower root powder.Getting fleece-flower root powder 10g packs in triangular flask, fleece-flower root powder is carried out to lixiviate with sterile water, aseptic moisture adds for 3 times, adds after 40mL at every turn, put triangular flask on shaking table at 27 DEG C with 115r/min jolting 48h, filter, collect filtrate, merge 3 times filtrate, filtrate is inserted to rotation concentration and evaporation instrument, be that 50 DEG C and condensation temperature are under 7 DEG C of conditions at 0.09MPa air pressure, bath temperature, 120r/min evaporation and concentration, to paste, obtains reddish yellow radix-polygoni multiflori extract.
Embodiment 3
Get after the fleece-flower root is cleaned and dry, be crushed to 95 order fine powders with the rotating speed of 5000r/min with high speed disintegrator, acquisition fleece-flower root powder.Getting fleece-flower root powder 10g packs in triangular flask, fleece-flower root powder is carried out to lixiviate with sterile water, aseptic moisture adds for 3 times, adds after 50mL at every turn, put triangular flask on shaking table at 29 DEG C with 130r/min jolting 36h, filter, collect filtrate, merge 3 times filtrate, filtrate is inserted to rotation concentration and evaporation instrument, be that 55 DEG C and condensation temperature are under 9 DEG C of conditions at 0.1MPa air pressure, bath temperature, 130r/min evaporation and concentration, to paste, obtains reddish yellow radix-polygoni multiflori extract.
The inhibition of example 4 radix-polygoni multiflori extracts to Botrytis cinerea germ
The radix-polygoni multiflori extract that takes above-mentioned preparation packs in triangular flask, and being diluted to concentration with sterile water 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 grey mould fruit rot of strawberry germ bacterium cake horizontalization plate central authorities that inoculation diameter is 5mm, do medicament contrast with carbendazim (98% carbendazim active compound is with obtaining after 0.1mol/L diluted hydrochloric acid dissolution) simultaneously, sterile water is made blank, every processing repeats 3 times, at 28-30 DEG C, 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 Botrytis cinerea germ
Note: different lowercases represent significant difference (p<0.05), 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 Botrytis cinerea germ.Indoor antibacterial test result shows: cultivating after 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, for control 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.First be the solution of 35mg/mL by radix-polygoni multiflori extract with sterile water dilution, simultaneously, Botrytis cinerea germ bacterium cake is placed on potato dextrose agar flat board and cultivates 96h, after conidium grows, diluting with sterile water is the spore suspension of 1 × 106/mL, the conidial suspension of getting equivalent mixes rear dripping on slide with the solution after aforementioned dilution, taking carbendazim as contrast medicament, and establish blank, repeat 3 times, in culture dish, 48h is cultivated in moisturizing, examine under a microscope 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
Note: different lowercases represent significant difference (p<0.05), 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, wherein radix-polygoni multiflori extract is processed Botrytis cinerea germ sclerotium germination rate lower than without radix-polygoni multiflori extract processing, but higher than the processing 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.Because conidium can be caused protection, grey mould fruit rot of strawberry infects again, causes the underproduction, therefore, suppresses conidia germination, thereby reduce source of infection again by using radix-polygoni multiflori extract, alleviates disease and occurs.
The impact that example 6 radix-polygoni multiflori extracts are sprouted Botrytis cinerea germ sclerotium
Botrytis cinerea germ is inoculated on potato dextrose agar flat board, cultivates 7d for 20 DEG C, when sclerotium blackening is ripe, collect sclerotium stand-by.Sclerotium is immersed in containing 48h in the solution of series concentration (0.1mg/mL, 10mg/mL, 20mg/mL, 40mg/mL, 80mg/mL) radix-polygoni multiflori extract, air-dry being placed on PDA flat board; To not be placed in containing on the PDA culture medium flat plate of above-mentioned series concentration radix-polygoni multiflori extract with the sclerotium of radix-polygoni multiflori extract processing, 20 DEG C of cultivations, every processing repeats 3 times, 100 sclerotium of each replication.Relatively the sclerotium of two kinds of processing is sprouted situation, measures sclerotium germination rate, the results are shown in following table 3.
Results and analysis: as shown in Table 3, process sclerotium germination rate through radix-polygoni multiflori extract and be starkly lower than the germination rate without radix-polygoni multiflori extract processing, and along with the rising of radix-polygoni multiflori extract concentration, sclerotium germination rate reduces gradually, wherein in the time that drug concentration is brought up to 80mg/mL, the germination rate of sclerotium is minimum, illustrates that radix-polygoni multiflori extract sprouts and play inhibitory action Microsclerotia.
The results of analysis of variance shows: the sclerotium germination rate significant difference of different radix-polygoni multiflori extract processing, but it is also remarkable to contrast sclerotium difference, and this may be not quite identical relevant with sclerotium size.Because sclerotium can be resisted unsuitable environmental condition, can help pathogen to cross poor environment, the present invention finds that radix-polygoni multiflori extract can reduce sclerotium germination rate after deliberation, 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 |
Note: different lowercases represent significant difference (p<0.05), different capitalizations represent utmost point significant difference (p<0.01).
The in vitro control efficiency of example 7 radix-polygoni multiflori extracts to 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 carry out surface sterilization 5s with 75% alcohol, then with washing out residual alcohol in sterile water, be placed in vent cabinet natural air drying.Get respectively radix-polygoni multiflori extract, carbendazim medicament, sterile water and be evenly applied in From Strawberry Leaves tow sides, be placed in vent cabinet natural air drying, buy the fresh bacterium cake of getting the botrytis cinerea colony edge having activated with the card punch of internal diameter 5mm, choose to blade with oese, when inoculation, avoid bacterium cake contact vein position, after smearing, blade is placed in culture dish, at temperature 26-28 DEG C, 3d is cultivated in moisturizing, 3 blades of every processing also repeat 3 times, adopt right-angled intersection method to measure strawberry fruit scab diameter, calculate control efficiency, the results are shown in following table 4.
Control efficiency (%)=[(contrast disease spot diameter – and process scab diameter)/(contrast scab diameter-5)] × 100%.
Results and analysis: as shown in Figure 1, radix-polygoni multiflori extract on From Strawberry Leaves eubolism without impact, and the From Strawberry Leaves gray mold scab diameter of radix-polygoni multiflori extract processing be significantly less than blank and process after ash arrhizus bacteria infect the scab diameter of formation, but be greater than the processing of contrast medicament carbendazim.By measuring 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., can be developed as biological prevention and control agent.
The in vitro control efficiency of table 4 radix-polygoni multiflori extract of the present invention to 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 |
| Sterile water contrast | 53.43 | 53.2 | 52.34 | 52.99 | ? |
Note: different lowercases represent significant difference (p<0.05), different capitalizations represent utmost point significant difference (p<0.01).
The in vitro control efficiency of example 8 radix-polygoni multiflori extracts to strawberry fruit gray mold
Win fruit size consistent, without the ripe strawberry fruit of damage by disease and insect, first carry out surface sterilization 3s with 75% alcohol, then use the alcohol of sterile water wash residual, be placed in vent cabinet natural air drying.Get radix-polygoni multiflori extract and smear at the aseptic absorbent cotton of the malicious fruit surface of grass, be placed in vent cabinet natural air drying.Wash down activating on the botrytis cinerea bacterium colony of 72h raw fresh botrytis cinerea spore with sterile water, be mixed with 1 × 10
6the concentration of individual/mL, spray the circular spot of 5mm at strawberry fruit base portion with watering can, 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 strawberry fruit eubolism without impact, and the ash arrhizus bacteria that the scab diameter that ash arrhizus bacteria infects formation after radix-polygoni multiflori extract is processed is significantly less than control treatment infects the scab diameter of formation, but be greater than the scab diameter of contrast medicament carbendazim.Meanwhile, measure scab diameter by right-angled intersection method, 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 in vitro control efficiency of table 5 radix-polygoni multiflori extract of the present invention to strawberry fruit gray mold
Note: different lowercases represent significant difference (p<0.05), different capitalizations represent utmost point significant difference (p<0.01).
Claims (2)
1. radix-polygoni multiflori extract, in an application of controlling in grey mould fruit rot of strawberry, is characterized in that the water extract that described radix-polygoni multiflori extract is the fleece-flower root is to be prepared from the steps below:
A, get polygonum multiforum root tuber clean after dry to water content be 1.5-4.5%, be crushed to 75-95 order fine powder through high speed disintegrator with the rotating speed of 3000-5000r/min;
B, fleece-flower root powder is carried out to lixiviate with sterile water, fleece-flower root powder is disposable to add, aseptic moisture adds for 3 times, after the ratio that at every turn adds 30-50mL sterile water in 1g fleece-flower root powder adds, on shaking table at 25-29 DEG C 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 DEG C and condensation temperature are that under 5-9 DEG C of condition, 110-130r/min is concentrated into paste at 0.08-0.1MPa air pressure, bath temperature.
2. the application of radix-polygoni multiflori extract in the in vitro From Strawberry Leaves gray mold of control and in vitro strawberry fruit gray mold, is characterized in that the water extract that described radix-polygoni multiflori extract is the fleece-flower root is to make and form in the steps below:
A, get polygonum multiforum root tuber clean after dry to water content be 1.5-4.5%, be crushed to 75-95 order fine powder through high speed disintegrator with the rotating speed of 3000-5000r/min;
B, fleece-flower root powder is carried out to lixiviate with sterile water, fleece-flower root powder is disposable to add, aseptic moisture adds for 3 times, after the ratio that at every turn adds 30-50mL sterile water in 1g fleece-flower root powder adds, on shaking table at 25-29 DEG C 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 DEG C and condensation temperature are that under 5-9 DEG C of condition, 110-130r/min is concentrated into paste at 0.08-0.1MPa air pressure, bath temperature.
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