CN108124905B - Application of fulvic acid in preventing and treating post-harvest gray mold of grape fruits - Google Patents
Application of fulvic acid in preventing and treating post-harvest gray mold of grape fruits Download PDFInfo
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- CN108124905B CN108124905B CN201810105475.5A CN201810105475A CN108124905B CN 108124905 B CN108124905 B CN 108124905B CN 201810105475 A CN201810105475 A CN 201810105475A CN 108124905 B CN108124905 B CN 108124905B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N61/00—Biocides, pest repellants or attractants, or plant growth regulators containing substances of unknown or undetermined composition, e.g. substances characterised only by the mode of action
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Abstract
The invention discloses application of fulvic acid in preventing and treating post-harvest gray mold of grape fruits. When the grape gray mold is prevented and treated, fulvic acid aqueous solution is adopted, the mass volume concentration of the fulvic acid aqueous solution is 0-20 mg/mL but not zero, and the content of polyphenol in the grape is specifically increased. The fulvic acid substance adopted by the invention can be used for preventing and treating the gray mold of picked grapes, can be used independently, and can also be used together with other active substances to enhance the antibacterial effect or reduce the toxic and side effect of the fulvic acid substance.
Description
Technical Field
The invention relates to an application of fulvic acid in prevention and treatment of post-harvest gray mold of grapes, and belongs to the technical field of biology.
Background
The grape is an important economic crop, the application of the grape relates to the aspects of wine making, fresh eating, dried fruits and the like, and the grape is closely related to the development of national economy and the health of human bodies. However, gray mold caused by Botrytis cinerea (Botrytis cinerea) is an important plant disease, which mainly damages grape spica and fruits on grapes, causes flower and fruit dropping and fruit rot, seriously affects postharvest storage and transportation of grapes, and limits the development of the grape industry.
At present, the post-harvest treatment of grapes is mainly carried out by SO2Fumigating to prevent and treat diseases and pestsInduced alteration of fruit quality and SO2The problems of harm to human bodies and the like caused by residues are not ignored, so that a novel alternative method for preventing and treating the botrytis cinerea after grape harvest is urgently required to be explored.
The induced resistance control of plants is mainly to improve the resistance of the crops to external stress and plant diseases and insect pests by spraying various inducers in vitro, and the common inducers comprise Si, chitosan, some biocontrol microbial inoculum and the like. Fulvic acid, a water-soluble substance in humic acid, is attracting attention in agricultural production because of its excellent properties of improving soil properties, increasing crop yield, enhancing crop resistance, etc. as a soil-applied fertilizer, among which common fulvic acid fertilizers include potassium fulvate and sodium fulvate. In addition, fulvic acid is used as a medicament auxiliary agent commonly used in the pharmaceutical industry, has certain curative effects on gastroenteritis, rheumatism, arthritis and the like, and is beneficial to human health.
Disclosure of Invention
The invention aims to provide application of fulvic acid in prevention and treatment of botrytis cinerea after grape harvest, wherein the fulvic acid can prevent and treat fruits infected by botrytis cinerea and has remarkable prevention and treatment effects on occurrence of fruit diseases and disease expansion.
The invention firstly provides the application of fulvic acid in preventing and treating post-harvest gray mold of grapes.
In the specific application, grapes before or after botrytis cinerea infection are soaked in fulvic acid aqueous solution;
the mass volume concentration of the fulvic acid aqueous solution can be 0-20 mg/mL, but is not zero, such as 10 mg/mL;
the fulvic acid can prevent and treat gray mold, and is specifically represented by increasing the content of polyphenol in grapes.
The invention further provides a method for preventing and treating the post-harvest gray mold of grapes, which comprises the following steps:
the grapes before or after the botrytis cinerea infection are soaked in the fulvic acid water solution, so that the control of the botrytis cinerea after the grapes are picked is realized;
the mass volume concentration of the fulvic acid aqueous solution can be 0-20 mg/mL, but is not zero, such as 10 mg/mL.
According to the method, the grape fruits are treated by selecting fulvic acid solutions with different concentrations, and then Botrytis cinerea is inoculated, and experimental results show that the gray mold occurrence degree of the grape fruits treated by the fulvic acid is remarkably reduced; in addition, the fulvic acid is selected to treat botrytis cinerea under the in vitro condition, so that the fulvic acid treatment has no influence on the growth of pathogenic bacteria, and the effect of preventing diseases is achieved by inducing the resistance of plants preliminarily; finally, the total polyphenol content of the grape fruits is extracted after the fulvic acid is used for treating the grape fruits, and the polyphenol content is found to be obviously improved. The method for preventing and treating the post-harvest gray mold of the grapes by the fulvic acid is considered to be feasible by combining the prevention effect of the fulvic acid, the bacteriostatic activity of the polyphenol and the benefit of human health.
The fulvic acid substance adopted by the invention can be used for preventing and treating the gray mold of picked grapes, can be used independently, and can also be used together with other active substances to enhance the antibacterial effect or reduce the toxic and side effect of the fulvic acid substance.
Drawings
FIG. 1 shows the control effect of fulvic acid on post harvest gray mold of grape fruits.
FIG. 2 is a graph showing the effect of fulvic acid on the growth of Botrytis cinerea (Botrytis cinerea) hyphae.
FIG. 3 is a graph of the effect of fulvic acid on the polyphenol content of grape fruits.
Detailed Description
The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Test pathogens used in the following examples: the Botrytis cinerea to be tested is separated into diseased grape fruits and is preserved on a potato dextrose solid medium (PDA).
The fulvic acid reagent used in the examples described below was obtained from mclin biochem technologies ltd and was 85% pure.
Example 1 Effect experiment of fulvic acid on prevention of post harvest grape gray mold
Test materials: the tested grape variety, mare's milk, was purchased from the market.
Manually separating single grape fruit, reserving pedicel, selecting healthy fruit particles with consistent size, shape and maturity, sterilizing in 2% sodium hypochlorite solution for 2min, washing with sterile water for 2 times, air drying, and performing grouping treatment.
(1) The grape fruits are respectively placed in fulvic acid solutions with different concentrations to be soaked for 10min, each concentration treatment is set to be 3 times, and each concentration treatment is set to be 30 fruits.
(2) Naturally airing the fruits, and placing all the fruits at 22 ℃ for 24 h.
(3) Culturing Botrytis cinerea on PDA culture medium for 5 days, washing the plate with sterile water, and shaking to obtain spore suspension (concentration of 3 × 10)4one/mL).
(4) The fruit was artificially punctured and 10. mu.L of Botrytis cinerea spore suspension was inoculated to the wound.
(5) The fruits processed at each concentration are respectively placed in a plastic preservation box, sprayed for moisture preservation and stored at 22 ℃.
(6) After storage for 4d and 6d, the incidence of fruit and lesion diameter were observed and recorded, and the inhibition was calculated. The inhibition ratio (%) is [ (a-B)/a ] × 100, a is the control group incidence/lesion diameter, and B is the treatment group incidence/lesion diameter.
The results of the experiment are shown in table 1 and fig. 1.
TABLE 1 Fulvic acid control effect on post-harvest grape gray mold
The experimental results show that: the fulvic acid has good prevention effect on grape gray mold, and the prevention effect is improved along with the increase of the concentration. When the test concentration is 10mg/mL, the inhibition rate of the fruit morbidity after 6 days of storage reaches 29.9%, and the inhibition effect of the fruit morbidity on the lesion diameter reaches 42.6%. When the concentration of the fulvic acid is 20mg/mL, the inhibition rate of the fulvic acid on the fruit morbidity after 6 days of storage reaches 61%, and the inhibition effect on the lesion diameter reaches 54%.
Example 2 Effect of fulvic acid on the growth of Botrytis cinerea hyphae
Dissolving fulvic acid in sterile deionized water, adding the dissolved fulvic acid into a PDA culture medium to prepare a fulvic acid-containing culture medium, and measuring the inhibition effect of fulvic acid with different concentrations on the growth of pathogenic bacteria hyphae by a hypha growth rate method. The method comprises the following specific steps:
dissolving fulvic acid by using sterile deionized water as a solvent to prepare a mother solution, then accurately adding a certain amount of fulvic acid solution into PDA culture medium melted at 50 ℃ respectively, and uniformly mixing to ensure that the final concentrations of fulvic acid in the culture medium respectively reach 0.1, 0.5, 1, 10 and 50mg/mL, setting 3 times of treatment for each concentration, and adding sterile deionized water with the same volume as a control. After culturing botrytis cinerea to be tested on a PDA culture medium for 5 days, a bacterial cake (d is 6mm) with consistent bacterial age and diameter is punched at the edge of an activated pathogenic bacteria colony by a sterilization puncher, the bacterial cake is reversely connected to the center of a drug-containing flat plate and cultured in a constant temperature incubator at 22 ℃, and the experimental result is regularly observed and recorded.
The experimental result is shown in figure 2, and the result in figure 2 shows that the fulvic acid does not influence the hypha growth of botrytis cinerea, and when the concentration of the fulvic acid is 50mg/mL, the hypha growth diameter of the botrytis cinerea is not obviously different from that of a control.
Example 3 Effect of Fulvic acid on the Polyphenol content of grape fruits
Dissolving fulvic acid in sterile deionized water to enable the final concentration of the fulvic acid to reach 10mg/mL, soaking the grape fruits in the fulvic acid solution for 10min, taking out the grape fruits, airing the grape fruits, and treating the grape fruits with the same volume of deionized water to serve as a test control. After 24 hours at 22 ℃ the grape fruits were inoculated with the botrytis cinerea spore suspension according to the method of example 1, incubated at 22 ℃ in the dark, and the treated grape fruits were sampled periodically at 12h intervals for the extraction and determination of the polyphenol content.
The method for extracting and measuring the total content of the grape peel polyphenol comprises the following steps: manually separating the peel from the sampled grape fruit, and draining the water with filter paperSeparately, the pericarp was snap frozen in liquid nitrogen and ground to powder, and 8mL of 80% methanol solution was added for polyphenol extraction. Shaking the extractive solution at 28 deg.C for 90min (200rpm), centrifuging at 4 deg.C (12,000g) for 10min, and collecting supernatant as crude extractive solution of polyphenol. Measuring total polyphenol content by adopting a folin-phenol colorimetric method, adding 184 μ L of water into 24 μ L of crude extract, adding 12 μ L of folin-phenol solution, standing at room temperature for 3min, adding 30 μ L of 20% Na2CO3The solution was allowed to stand at 40 ℃ for 2 hours, and then the absorbance was measured at 765 nm. And drawing a standard curve by using gallic acid as a standard sample, and further solving the total polyphenol content of the extracting solution.
The experimental result is shown in figure 3, and the result in figure 3 shows that the total polyphenol content of grape peel is increased by fulvic acid treatment, and the total polyphenol content is in an increasing trend along with the extension of the inoculation time of pathogenic bacteria; after the pathogenic bacteria are inoculated for 36 hours, the total content of polyphenol is highest under the treatment of fulvic acid.
Claims (2)
1. The application of fulvic acid in preventing and treating post-harvest gray mold of grapes;
immersing grapes before or after botrytis cinerea infection in a fulvic acid aqueous solution;
the mass volume concentration of the fulvic acid aqueous solution is 0-20 mg/mL but not zero;
said use is manifested by an increased content of polyphenols in said grapes.
2. A method for preventing and treating post-harvest gray mold of grapes comprises the following steps:
the grapes before or after the botrytis cinerea infection are soaked in the fulvic acid water solution, so that the control of the botrytis cinerea after the grapes are picked is realized;
the mass volume concentration of the fulvic acid aqueous solution is 0-20 mg/mL but not zero.
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