CN103392747B - Application of radix paeoniae alba extract - Google Patents
Application of radix paeoniae alba extract Download PDFInfo
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- CN103392747B CN103392747B CN201310374859.4A CN201310374859A CN103392747B CN 103392747 B CN103392747 B CN 103392747B CN 201310374859 A CN201310374859 A CN 201310374859A CN 103392747 B CN103392747 B CN 103392747B
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Abstract
The invention relates to an application of a radix paeoniae alba extract for controlling grey mould fruit rot of strawberry. The extract has a remarkable inhibiting effect and a certain in-vitro control efficiency to grey mould fruit rot of strawberry by way of inhibiting molecular spore germination, sclerotium germination and the like, and can be further developed to a novel plant source bactericide.
Description
Technical field
The invention belongs to biological technical field, relate generally to the application of radix paeoniae alba extraction, be specifically related to radix paeoniae alba extraction in the application of controlling in grey mould fruit rot of strawberry.
Background technology
Strawberry is not only of high nutritive value, but also can consolidate gums, fresh breath, moist throat, the effect of improving looks and building up resistance.In recent years, along with growth in the living standard, people also increase year by year to the consumption figure of strawberry, and this has brought vigor and vitality to the development of Strawberry industry, but because the culture technique of strawberry is had relatively high expectations, and be not easy to transport and shift, thereby producing region is more fixing, repeat for many years to plant strawberry in a place, easily accumulate pathogen, often cause Diseases of Strawberry occurrence and harm to have the trend of increasing the weight of, wherein grey mould fruit rot of strawberry is exactly one of Major Diseases of harm strawberry production.
Grey mould fruit rot of strawberry (Botrytis cinerea Pers.et Tris.) is a kind of Common Diseases of harm strawberry, this disease mainly causes fruit rot, the safety in production of strawberry is formed to serious threat, become the important factor in order of restriction strawberry volume increase.On producing, in order to control the occurrence and harm of grey mould fruit rot of strawberry, main by methods such as plantation disease-resistant variety, agricultural cultivation control, chemical control and biological controls.Because gray mold is a kind of facultative parasite, also there is the saprophytic phase, and also do not find now its antigen, breeding for disease resistance is difficult to carry out; The anti-rules for the treatment of of agricultural cultivation mainly, by strengthening the water and fertilizer management to field, regulate temperature, humidity and illumination etc. to reach the object that reduces disease, but it are not obvious to produce effects.Adopt chemical control to remain the Main Means of the careless malicious gray mold of China's control, but owing to using continuously chemical agent, particularly use continuously single medicament, cause gray mold germ to produce gradually pesticide resistance, the control efficiency of chemical pesticide also declines year by year, and pesticide spraying cannot be avoided being directly sprayed onto on fruit surface when dispenser, has polluted fruit, easily cause poisoning to occur.Nearest result of study shows, adopts that biological control grey mould fruit rot of strawberry not only can reach efficiently, safety, environmental protection, pollution-free, and effect is better, wherein from plant, find new bactericidal active substance, develop novel plant source bactericide, become one of focus of current people's concern.
The root of herbaceous peony is the dry root of the peeling of ranunculaceae plant Chinese herbaceous peony, and due to its cold nature, bitter, acid, return liver, the spleen channel, effect of the flat liver pain relieving of tool, nourishing blood for regulating menstruation, astringing YIN to stop sweating, can be used for that treatment headache is dizzy, hypochondriac pain, stomachache, four limbs contraction pain, and the deficiency of blood is sallow, irregular menstruation, spontaneous perspiration, night sweat etc.Existing result of study shows: the root of herbaceous peony also has anti-inflammatory, eases pain, protects the effect such as liver, immunological regulation.Up-to-date result of study demonstration, radix paeoniae alba extraction is to pathogen, as also inhibited in the malignant bacteria such as Escherichia coli, staphylococcus aureus.As can be seen here, the medical value of radix paeoniae alba extraction is very extensive, and studies to obtain also comparison system, but this research that is extracted in control agricultural plant disease aspect has no research.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, though the medicament using is inhibited to animal pathogen, may not be inhibited to plant pathogen.Forefathers' result of study shows: radix paeoniae alba extraction 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 root of herbaceous peony 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 application of a kind of radix paeoniae alba extraction 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 paeoniae alba extraction is at the application of controlling in grey mould fruit rot of strawberry, the acetone extract that this radix paeoniae alba extraction is the root of herbaceous peony.
Above-mentioned radix paeoniae alba extraction is to get root of herbaceous peony piece root to shine after dry grinding, White Peony Root is carried out to lixiviate with analysis pure acetone, gained filtrate evaporation and concentration and obtaining.Its concrete preparation method is as follows:
A, get root of herbaceous peony piece root dry to water content be 2.5-4.5%, be crushed to 95-115 order fine powder with the rotating speed of 6500-8500r/min.
B, with analyze pure acetone White Peony Root is carried out to lixiviate, White Peony Root is disposable to add, acetone divides 3 times and adds, after the ratio that at every turn adds 30-50mL acetone in 1g White Peony Root adds, on shaking table at 20-30 DEG C with 100-140r/min jolting 48-60h, 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 4-6 DEG C of condition, 110-150r/min is concentrated into paste at 0.08-0.1MPa air pressure, bath temperature.
Below be only a preparation method of the radix paeoniae alba extraction that exemplifies of the present invention, this extract can adopt prior art to be prepared in fact.
Radix paeoniae alba extraction of the present invention is by suppressing the modes such as conidia germination, Microsclerotia sprouting, inhibited to Botrytis cinerea germ, to preventing and treating in vitro From Strawberry Leaves and fruit gray mold has certain effect, and extracting method 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 paeoniae alba extraction to From Strawberry Leaves gray mold;
Wherein: 1 represents radix paeoniae alba extraction processing; 2 represent the processing of carbendazim liquid; 3 represent sterile water processing.
Fig. 2 is the in vitro control efficiency figure of radix paeoniae alba extraction to strawberry fruit gray mold;
Wherein: 1 represents radix paeoniae alba extraction processing; 2 represent the processing of carbendazim liquid; 3 represent sterile water processing.
Embodiment
Below in conjunction with concrete test and embodiment, technical scheme of the present invention is described further.
Raw material root of herbaceous peony piece root 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 25-28 DEG C.
Embodiment 1
Get after root of herbaceous peony piece root is cleaned and dry, be crushed to 95 order fine powders with the rotating speed of 6500r/min with Universalpulverizer, acquisition White Peony Root.Getting White Peony Root 10g packs in triangular flask, with analyzing pure acetone, White Peony Root is carried out to lixiviate, acetone divides 3 times and adds, and adds after 30mL at every turn, put triangular flask on shaking table at 20 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 4 DEG C of conditions at 0.08MPa air pressure, bath temperature, 110r/min evaporation and concentration, to paste, obtains yellowing look radix paeoniae alba extraction.
Embodiment 2
Get after the root of herbaceous peony is cleaned and dry, be crushed to 105 order fine powders with the rotating speed of 7500r/min with Universalpulverizer, acquisition White Peony Root.Getting White Peony Root 10g packs in triangular flask, with analyzing pure acetone, White Peony Root is carried out to lixiviate, acetone divides 3 times and adds, and adds after 40mL at every turn, put triangular flask on shaking table at 25 DEG C with 120r/min jolting 54h, 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 5 DEG C of conditions at 0.09MPa air pressure, bath temperature, 130r/min evaporation and concentration, to paste, obtains yellowing look radix paeoniae alba extraction.
Embodiment 3
Get after the root of herbaceous peony is cleaned and dry, be crushed to 115 order fine powders with the rotating speed of 8500r/min with Universalpulverizer, acquisition White Peony Root.Getting White Peony Root 10g packs in triangular flask, with analyzing pure acetone, White Peony Root is carried out to lixiviate, acetone divides 3 times and adds, and adds after 50mL at every turn, put triangular flask on shaking table at 30 DEG C with 150r/min jolting 48h, 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 6 DEG C of conditions at 0.1MPa air pressure, bath temperature, 150r/min evaporation and concentration, to paste, obtains yellowing look radix paeoniae alba extraction.
The inhibition of example 4 radix paeoniae alba extractions to Botrytis cinerea germ
The radix paeoniae alba extraction of getting above-mentioned preparation packs in triangular flask, be 40mg/mL solution with being diluted to concentration with sterile water again after a small amount of acetone solution, get this solution of 1mL and mix with 9mL potato dextrose agar (adding 30 μ g/mL streptomycins), shake up and make 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 (sterile water dilution after a small amount of 0.1mol/L diluted hydrochloric acid dissolution for 98% carbendazim active compound) simultaneously, sterile water is made blank, every processing repeats 3 times, at 28-30 DEG C, cultivate, after 48h, adopt right-angled intersection method to 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%.
Results and analysis: radix paeoniae alba extraction of the present invention has certain inhibition to Botrytis cinerea germ.Indoor antibacterial test result shows: cultivating after 48h, radix paeoniae alba extraction of the present invention is 61.92% to Botrytis cinerea germ inhibiting rate, suitable with the inhibition that contrasts medicament carbendazim.The results of analysis of variance also shows: radix paeoniae alba extraction is processed and contrast medicament carbendazim is processed without significant difference (table 1).Illustrate that radix paeoniae alba extraction of the present invention has the potentiality of further exploitation bactericide.
The inhibitory action of table 1 radix paeoniae alba extraction 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).
The impact of example 5 radix paeoniae alba extractions on the spore germination of grey mould fruit rot of strawberry bacterium molecule
Adopt slide sprouting method to detect the impact of radix paeoniae alba extraction on Botrytis cinerea germ conidia germination.First be 35mg/mL solution with sterile water dilution again with a small amount of acetone solution radix paeoniae alba extraction, meanwhile, Botrytis cinerea germ bacterium cake be placed on potato dextrose agar flat board and cultivate 96h, after conidium grows, diluting with sterile water is 1 × 10
6the spore suspension of individual/mL, get the conidial suspension of equivalent and the solution of aforementioned dilution and mix rear dripping on slide, taking carbendazim as contrast medicament, and establish blank, repeat 3 times, in culture dish, 48h is cultivated in moisturizing, examines under a microscope conidia germination situation, calculate spore germination rate, see the following form 2.
The impact of table 2 radix paeoniae alba extraction 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: the germination rate through the conidia germination rate of radix paeoniae alba extraction processing lower than blank, but higher than the germination rate of contrast medicament carbendazim processing, wherein the conidia germination rate of radix paeoniae alba extraction processing is 0.43 times of blank, but contrasts 1.41 times of medicament.The results of analysis of variance shows: radix paeoniae alba extraction is processed and contrasted between medicament carbendazim processing and the processing of blank not only significant difference, and reaches utmost point significance level.Illustrate that radix paeoniae alba extraction can reach the pathogenetic object of inhibition Botrytis cinerea by suppressing conidia germination.
The impact that example 6 radix paeoniae alba extractions are sprouted Botrytis cinerea germ sclerotium
Botrytis cinerea germ is inoculated on potato dextrose agar flat board, cultivates 7d for 20-30 DEG C, when sclerotium blackening is ripe, collect sclerotium stand-by.Sclerotium is immersed in containing 48h in the solution of series concentration (0.5mg/mL, 5mg/mL, 10mg/mL, 20mg/mL, 40mg/mL) radix paeoniae alba extraction, air-dry being placed on potato dextrose agar flat board; To not be placed in containing on the potato dextrose agar culture medium flat plate of above-mentioned series concentration radix paeoniae alba extraction with the sclerotium of radix paeoniae alba extraction processing, 20-30 DEG C of cultivation, 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, sees the following form 3.
Results and analysis: as shown in Table 3, after radix paeoniae alba extraction is processed, sclerotium is sprouted and is affected, and is wherein starkly lower than the sclerotium germination rate without radix paeoniae alba extraction processing through the sclerotium germination rate of radix paeoniae alba extraction processing.Different radix paeoniae alba extraction concentration and sclerotium germination rate are inverse ratio, and, along with the raising of radix paeoniae alba extraction concentration, the germination rate of different disposal sclerotium reduces gradually.Illustrate that radix paeoniae alba extraction sprouts and play inhibitory action Microsclerotia.Be not inoculated on the culture medium flat plate that contains variable concentrations radix paeoniae alba extraction with the sclerotium of radix paeoniae alba extraction processing, between each processing, the sprouting situation of sclerotium also has certain difference, and this may vary in size relevant with sclerotium.In addition, in the time that concentration is 40mg/mL, the growth of Botrytis cinerea germ mycelia is obviously suppressed, and along with the raising of radix paeoniae alba extraction concentration, inhibition is better.Therefore, radix paeoniae alba extraction not only suppresses the sprouting of sclerotium, and suppresses mycelial growth.
The impact that table 3 radix paeoniae alba extraction is sprouted Botrytis cinerea germ sclerotium
| Concentration (mg/mL) | Process sclerotium germination rate (%) | Untreated sclerotium germination rate (%) |
| 0.5 | 79.14aA | 96.61aA |
| 5 | 73.48bB | 96.40aA |
| 10 | 71.15cC | 95.23bB |
| 20 | 68.45dD | 95.16bBC |
| 40 | 64.7eE | 94.41cC |
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 paeoniae alba extractions 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 4s 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, see the following form 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 paeoniae alba extraction is on From Strawberry Leaves normal growth without impact, and the scab diameter that ash arrhizus bacteria infects formation after radix paeoniae alba extraction is processed is significantly less than control treatment, but is greater than the processing of contrast medicament carbendazim.By measuring colony diameter, calculate control efficiency and find that radix paeoniae alba extraction can reach 46.28% to the in vitro control efficiency of From Strawberry Leaves gray mold, lower than the processing of contrast medicament carbendazim.The results of analysis of variance also shows, radix paeoniae alba extraction process and contrast medicament carbendazim to process difference also remarkable.Some explanation radix paeoniae alba extraction can be used to prevent and treat grey mould fruit rot of strawberry above, and can be by improving concentration or compositely can reaching the object of controlling better grey mould fruit rot of strawberry.
The in vitro control efficiency of table 4 radix paeoniae alba extraction of the present invention to From Strawberry Leaves gray mold
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 paeoniae alba extractions 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 4s 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, in table 5.Control efficiency (%)=[(contrast colony diameter-processing colony diameter)/(contrast colony diameter-5)] × 100
Results and analysis: as shown in Figure 2, radix paeoniae alba extraction on strawberry fruit normal growth without impact, and get after thing is processed strawberry fruit is had to certain in vitro control efficiency through the root of herbaceous peony, wherein, be less than the processing of blank through the From Strawberry Leaves gray mold scab diameter of radix paeoniae alba extraction processing, but be greater than the processing of contrast medicament carbendazim, simultaneously, measure scab diameter by right-angled intersection method, calculate control efficiency find radix paeoniae alba extraction 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 paeoniae alba extraction is processed and contrast medicament carbendazim is processed significant difference (table 5).Illustrate that radix paeoniae alba extraction can be used for preventing and treating grey mould fruit rot of strawberry, and can be by improving radix paeoniae alba extraction concentration or compositely can reaching the object of controlling better grey mould fruit rot of strawberry.
The in vitro control efficiency of table 5 radix paeoniae alba extraction 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 paeoniae alba extraction, in an application of controlling in grey mould fruit rot of strawberry, is characterized in that, described radix paeoniae alba extraction is the acetone extract of the root of herbaceous peony, is to follow these steps to be prepared from:
A, get root of herbaceous peony piece root dry to water content be 2.5-4.5%, be crushed to 95-115 order fine powder;
B, with analyze pure acetone White Peony Root is carried out to lixiviate, White Peony Root is disposable to add, acetone divides 3 times and adds, after the ratio that at every turn adds 30-50mL acetone in 1g White Peony Root adds, on shaking table at 20-30 DEG C with 100-140r/min jolting 48-60h, filter, collect filtrate, merge 3 times filtrate;
C, gained filtrate is inserted to rotation concentration and evaporation instrument, be concentrated into paste.
2. the application of radix paeoniae alba extraction in the in vitro From Strawberry Leaves gray mold of control and in vitro strawberry fruit gray mold, is characterized in that, described radix paeoniae alba extraction is the acetone extract of the root of herbaceous peony, is to follow these steps to be prepared from:
A, get root of herbaceous peony piece root dry to water content be 2.5-4.5%, be crushed to 95-115 order fine powder;
B, with analyze pure acetone White Peony Root is carried out to lixiviate, White Peony Root is disposable to add, acetone divides 3 times and adds, after the ratio that at every turn adds 30-50mL acetone in 1g White Peony Root adds, on shaking table at 20-30 DEG C with 100-140r/min jolting 48-60h, filter, collect filtrate, merge 3 times filtrate;
C, gained filtrate is inserted to rotation concentration and evaporation instrument, be concentrated into paste.
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| CN102792977A (en) * | 2011-05-25 | 2012-11-28 | 杨松 | Method for preparing pesticide for preventing and controlling aphids of cherry tomatoes |
| CN102805112A (en) * | 2011-05-30 | 2012-12-05 | 王光达 | Botanical pesticide for controlling cucumber bacterial angular leaf spot |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102792977A (en) * | 2011-05-25 | 2012-11-28 | 杨松 | Method for preparing pesticide for preventing and controlling aphids of cherry tomatoes |
| CN102805112A (en) * | 2011-05-30 | 2012-12-05 | 王光达 | Botanical pesticide for controlling cucumber bacterial angular leaf spot |
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| "13 种药用植物粗提物抑菌活性的初步研究";于九伶等;《北京农学院学报》;20110415;第26卷(第2期);第11-12页 * |
| 于九伶等."13 种药用植物粗提物抑菌活性的初步研究".《北京农学院学报》.2011,第26卷(第2期),第11-12页. * |
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