CN115956564A - Application of tris Yang mycin in preventing and controlling pathogenic bacteria and application in plant preservation - Google Patents
Application of tris Yang mycin in preventing and controlling pathogenic bacteria and application in plant preservation Download PDFInfo
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Abstract
The invention discloses application of tri-Yang mycin in preventing and controlling pathogenic bacteria and plant preservation, and belongs to the technical field of pesticides. The invention aims to use the tri-Yang mycin in inhibiting the growth of the pathogenic fungus colletotrichum gloeosporioides of the postharvest citrus anthracnose and the postharvest fresh-keeping of citrus, and provides the application of the derivative of the tri-Yang mycin or the tri-Yang mycin in at least one of the following items, (1) the prevention and treatment of pathogenic bacteria; (2) increasing the resistance of plants to pathogenic bacteria; (3) inhibiting the pathogenicity of pathogenic bacteria; (4) inhibiting the hyphal growth of pathogenic bacteria; (5) improving the freshness of the fruits; the low-concentration tri-Yang mycin can effectively inhibit the growth of colletotrichum gloeosporioides, provides a new choice for preventing and controlling the anthracnose of citrus, provides a very meaningful basis for new drugs and choices of citrus preservative and fresh-keeping agents, and has great potential application value in the aspects of reducing the use of harmful chemical fresh-keeping agents and green sustainable development of storage and fresh-keeping of citrus.
Description
Technical Field
The invention belongs to the technical field of pesticides, and particularly relates to application of tris Yang mycin in preventing and controlling pathogenic bacteria and application in plant preservation.
Background
Citrus anthracnose is one of the most common diseases of postharvest citrus, and the pathogenic bacterium Colletotrichum gloeosporioides belongs to Ascomycota, chaetoceromycetes, cong Ke mesh, cong Keke, colletotrichum. The disease invades citrus fruits through wounds or is infected by mutual contact of diseased fruits and healthy fruits, resulting in a large number of rotten fruit cases. The disease frequently occurs in the world, the infectivity is extremely high, if the management is not good, the disease can be spread in a large area in a short time, even the whole warehouse is completely infected, and huge loss is caused to farmers and the whole industry.
At present, the main prevention and treatment means is chemical prevention and treatment. The traditional chemical pesticides such as benomyl, thiophanate methyl, mancozeb and the like all cause the citrus to generate larger drug resistance due to the continuous use for many years, and the control effect is poorer. At present, benomyl is internationally used for preventing and treating the anthracnose of the citrus, can effectively prevent and treat the anthracnose of the citrus, and is favored by vast citrus fruit growers. However, if a large amount of single pesticide varieties are used for a long time, the control effect is reduced, and the generation of drug resistance is easily caused.
Disclosure of Invention
The invention aims to use the tri-Yang mycin in inhibiting the growth of the pathogenic fungus Colletotrichum gloeosporioides (Colletotrichum gloeosporioides) of the citrus anthracnose after harvest and the fresh keeping of the citrus after harvest. Provides a novel pesticide component which can prevent and control pathogenic bacteria and has the effect of keeping fruits fresh.
The invention provides an application of a derivative of tri-Yang mycin or tri-Yang mycin in at least one of the following items,
(1) Preventing and treating pathogenic bacteria;
(2) Increasing the resistance of plants to pathogenic bacteria;
(3) Inhibiting the pathogenicity of pathogenic bacteria;
(4) Inhibiting the growth of hyphae of pathogenic bacteria;
(5) The fruit preservation is improved;
the molecular structural formula of the tri-Yang mycin is as follows:
further defined, the pathogenic bacteria are Colletotrichum gloeosporioides (Colletotrichum gloeosporioides), erysiphe necator (Sphaerotheca fuliginea), botrytis cinerea (Botrytis cinerea), cladosporium fulvum (Cladosporium fulvum), sclerotinia sclerotiorum (sclerotiotinia sclerotiorum), verticillium dahliae (Verticillium dahliae), phomopsis vexans (Phomopsis vexans), epispermum (diaporhe vexans), oxysporum (Fusarium oxysporum), rhizoctonia solani (Rhizoctonia solani), and anthracnose pathogenic bacteria (c.
The invention provides an application of a derivative of tri-Yang mycin or tri-Yang mycin in preparing at least one of the following functional products,
(1) Preventing and treating pathogenic bacteria;
(2) Improving the resistance of plants to pathogenic bacteria;
(3) Inhibiting the pathogenicity of pathogenic bacteria;
(4) Inhibiting the growth of hyphae of pathogenic bacteria;
(5) The fruit preservation is improved;
the molecular structural formula of the tri-Yang mycin is as follows:
further defined, the functional product is a pesticide.
Further defined, the pathogenic bacteria are Colletotrichum gloeosporioides (Colletotrichum gloeosporioides), erysiphe necator (Sphaerotheca fuliginea), botrytis cinerea (Botrytis cinerea), cladosporium fulvum (Cladosporium fulvum), sclerotinia sclerotiorum (sclerotirotifera), verticillium dahliae (Verticillium dahliae), phomopsis fungs (Phomopsis vexans), aschersonia piperita (diaphoras vexans), neurospora oxysporum (Fusarium oxysporum), rhizoctonia solani (Rhizoctonia solani), anthrax pathogenic bacteria (c.
The invention provides a product containing derivatives of tri-Yang mycin or tri-Yang mycin, wherein the active ingredient of the product is the derivatives of tri-Yang mycin or tri-Yang mycin, and the product has the functions of at least one of the following:
(1) Preventing and treating pathogenic bacteria;
(2) Increasing the resistance of plants to pathogenic bacteria;
(3) Inhibiting the pathogenicity of pathogenic bacteria;
(4) Inhibiting the growth of hyphae of pathogenic bacteria;
(5) The fruit preservation is improved;
the molecular structural formula of the tri-Yang mycin is as follows:
further defined, the pathogenic bacteria are Colletotrichum gloeosporioides (Colletotrichum gloeosporioides), erysiphe necator (Sphaerotheca fuliginea), botrytis cinerea (Botrytis cinerea), cladosporium fulvum (Cladosporium fulvum), sclerotinia sclerotiorum (sclerotirotifera), verticillium dahliae (Verticillium dahliae), phomopsis fungs (Phomopsis vexans), aschersonia piperita (diaphoras vexans), neurospora oxysporum (Fusarium oxysporum), rhizoctonia solani (Rhizoctonia solani), anthrax pathogenic bacteria (c.
The invention provides a method for improving anthracnose resistance of citrus, which comprises the following specific steps: citrus is treated with an agent comprising a derivative of either tri Yang or tri Yang mycin.
Further defined, the reagent is a solution containing 10. Mu.g/mL of tris Yang mycin.
The invention provides a method for improving the fresh keeping of oranges, which comprises the following specific steps: citrus is treated with an agent comprising a derivative of either tri Yang or tri Yang mycin.
Further defined, the reagent is a solution containing 10 μ g/mL of tris Yang mycin and the treatment is dipping the citrus fruit in the reagent.
Has the advantages that: the invention uses the tri-Yang mycin for the prevention and treatment of the citrus anthracnose for the first time, in an in-vivo experiment of the citrus, the 10 microgram/mL tri-Yang mycin can effectively inhibit the growth of colletotrichum gloeosporioides, provides a new choice for the prevention and treatment of the citrus anthracnose, provides a very meaningful basis for the new medicine and the choice of the citrus preservative, and has great potential application value in the aspects of reducing the use of harmful chemical preservatives and the green sustainable development of the storage and the preservation of the citrus. The invention provides a bactericidal and/or bacteriostatic medicine, which is tri-Yang mycin. The drug has better control effect on the anthracnose of the citrus than the current main control drug benomyl, delays the generation of drug resistance of the citrus, is more environment-friendly, and has the function of a preservative for postharvest storage of the citrus. The tri-Yang mycin (Sanyangmycin) has the advantages of low toxicity, high efficiency, broad spectrum of action, environmental friendliness and the like, and almost has no cytotoxicity. The mouse is injected with 2.8g/kg in the abdominal cavity without any toxic and side effects.
Drawings
FIG. 1 shows the ITS sequence of pathogenic bacterium colletotrichum gloeosporioides obtained by PCR amplification.
FIG. 2 shows the structure of tris Yang mycin.
FIG. 3 is a graph of the inhibitory effect of tris Yang mycin on colletotrichum gloeosporioides.
FIG. 4 is a graph of the in vivo antibacterial effect of tris Yang mycin.
FIG. 5 shows lesion diameter (mm) and incidence (%) 3-9d after inoculation treatment.
FIG. 6 is a graph of the effect of tris Yang mycin on respiration intensity, weight loss rate and titratable acid content during postharvest storage of citrus fruit.
Detailed Description
The present invention will be described in detail with reference to specific examples. The following examples will assist those skilled in the art in further understanding the invention, but are not intended to limit the invention in any way. It should be noted that variations and modifications can be made by persons skilled in the art without departing from the concept of the invention. All falling within the scope of the present invention.
The raw materials used in the following examples were commercially available without specific indication.
1. Tris Yang mycin (Sanyangmycin): is a chemical reagent available from Aladdin reagent company, CAS number 28047-05-4, molecular formula C 12 H 15 NO 4 The common names are: acetyl-O-methyl-L-tyrosine, the structural formula is shown in figure 2.
2. Colletotrichum gloeosporioides strain (Colletotrichum gloeosporioides): obtained by separation in this experiment.
3. Potato dextrose agar medium (PDA): 200g of potatoes are cleaned, cut into small pieces, boiled for about 30min and filtered by four layers of gauze, the filtrate is replenished to 1000mL by tap water, and 20g of glucose and 20g of agar are added.
The pathogenic bacteria of citrus anthracnose are Colletotrichum gloeosporioides (Colletotrichum gloeosporioides) belonging to the phylum Ascomycota (ascota), class coprinus (sortarionomycetes), order Cong Ke (glomerilles), order Cong Keke (glomerilleae), genus Colletotrichum (Colletotrichum).
Colletotrichum gloeosporioides is pathogenic bacteria of citrus anthracnose;
and (3) safety detection: the tri-Yang mycin (Sanyangmycin) has the advantages of low toxicity, high efficiency, broad spectrum of action, environmental friendliness and the like, and almost has no cytotoxicity. 2.8g/kg of the compound is injected into the abdominal cavity of the mouse, and no toxic or side effect is caused.
Example 1 Tri Yang mycin inhibits the growth of the hyphae of the pathogenic bacterium colletotrichum
1. The separation method of colletotrichum gloeosporioides comprises the following steps: and separating and purifying pathogenic bacteria on the citrus infected with anthracnose after citrus is picked from a citrus orchard in a Oghai region of Wenzhou city of three Yang wetland to obtain the colletotrichum gloeosporioides.
2. ITS general primers ITS1 (5'-TCCGTAGGTGAACCTGCGG-3', SEQ ID NO. 1) and ITS4 (5'-TCCTCCGCTTATTGATATGC-3', SEQ ID NO. 2) are used for amplifying an ITS sequence (sequence: CTGCGGAGGGATCATTACTGAGTTTACGCTCTACAACCCTTTGTGAACATACCTACAACTGTTGCTTCGGCGGGTAGGGTCTCCGCGACCCTCCCGGCCTCCCGCCTCCGGGCGGGTCGGCGCCCGCCGGAGGATAACCAAACTCTGATTTAACGACGTTTCTTCTGAGTGGTACAAGCAAATAATCAAAACTTTTAACAACGGATCTCTTGGTTCTGGCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATTCAGTGAATCATCGAATCTTTGAACGCACATTGCGCCCGCCAGCATTCTGGCGGGCATGCCTGTTCGAGCGTCATTTCAACCCTCAAGCTCTGCTTGGTGTTGGGGCCCTACAGCCGATGTAGGCCCTCAAAGGTAGTGGCGGACCCTCCCGGAGCCTCCTTTGCGTAGTAACTTTACGTCTCGCACTGGGATCCGGAGGGACTCTTGCCGTAAAACCCCCCAATTTTCCAAAGGTTGACCTCGGATCAGGTAGGAATACCCGCTGAACTTAAGCATA, SEQ ID NO. 3), pathogenic fungi of the collected citrus anthracnose are identified, the separated pathogenic bacteria are confirmed to be colletotrichum gloeosporioides, the result is shown in figure 1, citrus tissue blocks with serious anthracnose infection characteristics are separated to a potato glucose culture medium for culture, and after multiple passages and purification, single pathogenic bacteria are separated on a culture plate.
650 base fragments were obtained by PCR amplification of ITS sequences from pathogenic fungi (see FIG. 1), and these base sequences were obtained by sequencing. And the comparison of an NCBI database proves that the separated pathogenic fungi is colletotrichum gloeosporioides.
(1) Activating strains: inoculating the colletotrichum gloeosporioides strain separated in the earlier stage experiment into a PDA culture medium, culturing in an incubator at 28 ℃, and activating for later use.
(2) Preparing spore suspension: the cultured colletotrichum gloeosporioides strain was taken, and 2mL of sterile water was added to the culture plate. Gently shaking to elute conidia. Sucking 1mL of the eluted spore suspension with a pipette and loading the spore suspension into a 1.5mL EP tube, and diluting the spore suspension to a concentration of 1X 10 5 spores/mL, stored in a refrigerator at 4 ℃ for later use.
3. Bacteriostatic test by a filter paper method: taking a sterilized filter paper sheet, and dripping 10 mu L of the tri-Yang mycin solution with the concentration of 5 mu g/mL,10 mu g/mL,15 mu g/mL and 20 mu g/mL on the filter paper sheet. 10 μ L of sterile water was used as a blank control, and an equivalent optimal concentration (10 μ g/mL) of benomyl was used as a positive control. And after the filter paper sheets are naturally dried, placing the filter paper sheets at positions 2.5cm away from the left and right of the colletotrichum gloeosporioides cake. The filter paper sheets were added with equal amounts of sterile water as controls, and each treatment was repeated 3 times and incubated at 28 ℃ in the dark. When the pathogenic bacteria of the sterile water control group grow over the dish, respectively measuring the diameter of the pathogenic bacteria colony of the control group and the diameter of the pathogenic bacteria colony expanding towards the direction of the filter paper sheet, and calculating the bacteriostasis rate according to the following formula:
hypha growth inhibition rate (%) = [ (control group pathogen diameter-treatment group pathogen diameter)/control group pathogen diameter ] × 100.
Results as shown in table 1 and fig. 3, the tris Yang mycin showed significant inhibitory effect against the anthrax pathogen (c. Wherein, under the concentration of 10 mug/mL, the inhibition rate of the three Yang mycin on the anthrax germ is the best, 81.4 percent, which is superior to the positive drug benomyl with the same concentration, and the inhibition rate of the benomyl on the anthrax germ is only 59.5 percent.
TABLE 1 statistics of the bacteriostatic rate of different concentrations of tris Yang mycin on anthrax pathogenic bacteria
Example 2 application of Tri Yang mycin to improvement of anthracnose resistance of citrus
Pricking a diameter of 5mm at the waist position of citrus fruit with sterile needle 2 20. Mu.L of the solution of the tri-Yang mycin with the concentrations of 5. Mu.g/mL, 10. Mu.g/mL, 15. Mu.g/mL and 20. Mu.g/mL was pipetted by a pipette and injected into the well. 20 μ L of sterile water was used as a blank control, and an equivalent optimal concentration (10 μ g/mL) of benomyl was used as a positive control.
After standing at 25 ℃ for 4h, 10. Mu.L of the suspension was added at a concentration of 1X 10 5 Spors/mL of spore suspension of colletotrichum gloeosporioides was injected into each wound. The inoculated citrus fruits are covered by a preservative film, stored in a constant-temperature incubator at 25 ℃ and cultured in the dark. The lesion diameter and incidence were recorded daily. Each antagonistic strain was subjected to 3 experiments, each treatment was repeated 3 times. The incidence was calculated according to the following formula: incidence (%) = (number of rotten wounds/total number of wounds) × 100%.
Incubate at 28 ℃ for 9d as shown in FIGS. 4 and 5. The Control group (Control) not inoculated with pathogenic bacteria showed normal fruit wound without signs of disease. The group inoculated with the pathogen alone (In) showed the most severe symptoms with large areas of irregular dark brown lesions, and the group inoculated with the tris Yang mycin all significantly reduced the incidence and lesion diameter (as In table 2). Wherein, the group (In + S) treated by the tri-Yang mycin with the concentration of 10 mug/mL has the best disease resistance, the disease symptoms are the lightest, the disease spots appear later, the disease incidence is reduced by 55.2 percent, and the diameter of the disease spots is reduced by 65.4 percent. The incidence rate of the benomyl inoculated group (In + B) is only reduced by 37.5 percent, and the diameter of the lesion spot is reduced by 29.8 percent. The data show that the protective effect of the tri-Yang mycin is obvious (P is less than or equal to 0.05) and is better than that of a positive control benomyl.
TABLE 2 Difference in biocontrol effects of citrus anthracnose with different concentrations of tris Yang
| Tri Yang Myomycin concentration (μ g/mL) | Incidence (%) | Diameter of lesion (mm) |
| 0 | 100 | 70.1±5.6 |
| 5 | 67.7±2.3 | 38.3±4.3 |
| 10 | 55.2±1.2 | 24.2±2.8 |
| 15 | 54.7±2.5 | 28.9±3.5 |
| 20 | 53.1±1.9 | 27.5±4.1 |
In addition, the tri-Yang mycin has a remarkable control effect on anthracnose after the loquat, mango and banana are picked, the morbidity of the loquat can be reduced by 32.7%, the diameter of a disease spot can be reduced by 41.5%, the morbidity of the mango can be reduced by 47.3%, the diameter of the disease spot can be reduced by 28.8%, the morbidity of the strawberry can be reduced by 39.1%, and the diameter of the disease spot can be reduced by 27.4%.
Example 3 broad-spectrum antibacterial Effect of Tri Yang Myomycin against various plant pathogens
Determination of Minimum Inhibitory Concentration (MIC) of tris Yang, mycin against various phytopathogens: as a control group 1, 200. Mu.L of PDB medium was added to a 96-well plate. As a control group 2, 100. Mu.L each of spore suspension of colletotrichum gloeosporioides and PDB medium was added. The experimental group was prepared by sequentially adding 100. Mu.L of spore suspension of pathogenic fungi to each blank well, and then adding 100. Mu.L each of positive control cycloheximide and tris Yang mycin. The final concentration gradient of the drug is adjusted to be 128, 64, 32, 16, 8, 4, 2, 1, 0.5 and 0.25 mu g/mL. Each gradient was repeated three times and incubated in the dark at 28 ℃ for 48h. The turbidity in each well plate was visually observed and the MIC value was preliminarily determined. Then the culture solution was discarded, and 100. Mu.L of MTT-PDB culture solution was added to each well, which was then cultured in an incubator at 28 ℃ for 4 hours. Wash once by adding 1 XPBS, then add 150. Mu.L DMSO solution per well. After incubation for 10min, the absorbance value is measured at the wavelength of 570nm, and the bacteriostasis rate is calculated. As shown in table 3, the results of the test results show that the tri-Yang mycin showed strong inhibitory effect on all 10 plant pathogens including anthrax pathogen (c). The tri-Yang mycin can be used as a plant pesticide.
TABLE 3 minimum inhibitory concentrations of Tri Yang mycin against several common pathogenic fungi
Example 4 application of Tris Yang mycin in the preservation of citrus after harvest
1. Coating treatment of citrus fruits: dissolving tris Yang mycin in water, stirring thoroughly until it is mixed uniformly, and preparing into solution with a concentration of 10 μ g/mL. Soaking the washed and air-dried citrus fruits in the solution, performing tri-Yang mycin coating treatment, and drying to form a film on the surfaces of the citrus fruits. Storing the treated citrus fruit at 25 ℃. Distilled water was used as a blank control and benomyl coating was used as a positive control.
2. Respiration intensity (CO) of citrus fruit 2 Release amount) detection: five fruits were sealed in closed 2.6L jars at ambient temperature (25 ℃) for 3h, with 3 biological replicates per group. Headspace gas samples were extracted from each vessel using a closed syringe and then injected into a gas chromatograph (7890a, aglent, usa). Respiratory intensity with CO 2 The amount of production, expressed as ml kg 1 ·h 1 . The measurement conditions were as follows: the column was packed with CTR 1 at 40 ℃ and the TCD detector at 120 ℃ and each treatment was repeated 3 times.
3. And (3) detecting the weight loss rate of the citrus fruit: each group of 20 fruits was used for weight loss testing and weighed every 5 days during storage. The results are shown as percent loss from the initial weight.
4. Titratable acids were measured by acidimeter (GMK 835, g WON Hitech, korea) according to the instructions for use of the instrument, and the results were expressed as percentages.
As shown in fig. 6, after treatment with the tri-Yang, the respiration rate and weight loss rate of citrus were significantly lower than those of the distilled water control group and the benomyl treatment group. The titratable acid was significantly higher than the distilled water control and benomyl treated groups. It is shown that treatment with the tri-Yang mycin greatly reduced the respiration rate of citrus fruit during storage, retained moisture and titratable acid levels, and had the best fruit quality at the end of the storage period. Therefore, the tri-Yang mycin coating can improve the fresh-keeping efficiency of the citrus fruit and can be used for prolonging the storage period of the citrus fruit after picking.
In addition, the tri-Yang mycin delays the fruit senescence and spoilage process by reducing respiration, delaying weight loss and maintaining the content of Titratable Acid (TA) of flavor substances, and significantly prolongs the postharvest storage period of loquat, mango and strawberry. The tri-Yang mycin can be used as a plant pesticide.
Claims (10)
1. Use of a derivative of tris Yang mycin or tris Yang mycin in at least one of,
(1) Preventing and treating pathogenic bacteria;
(2) Improving the resistance of plants to pathogenic bacteria;
(3) Inhibiting the pathogenicity of pathogenic bacteria;
(4) Inhibiting the hypha growth of pathogenic bacteria;
(5) The fruit preservation is improved;
the molecular structural formula of the tri-Yang mycin is as follows:
2. use according to claim 1, characterized in that the pathogenic bacteria are Colletotrichum gloeosporioides (Colletotrichum gloeosporioides), sphaerotheca fuliginea (Sphaerotheca fuliginea), botrytis cinerea (Botrytis cinerea), cladosporium fulvidrum (Cladosporium fulvum), sclerotinia sclerotiorum (Sclerotinia sclerororium), verticillium dahliae (Verticillium flaiae), phomopsis vexans (Phomopsis vexans), aschersonia metacarpus (Diabrospirhe vexans), fusarium oxysporum (Fusarium oxysporum), rhizoctonia solani (Rhizoctonia solani), anthrax (C.
3. The application of the derivatives of the three Yang mycin or the three Yang mycin in the preparation of at least one of the following functional products,
(1) Preventing and treating pathogenic bacteria;
(2) Improving the resistance of plants to pathogenic bacteria;
(3) Inhibiting the pathogenicity of pathogenic bacteria;
(4) Inhibiting the hypha growth of pathogenic bacteria;
(5) The fruit preservation is improved;
the molecular structural formula of the tri-Yang mycin is as follows:
4. use according to claim 3, characterized in that the pathogenic bacteria are Colletotrichum gloeosporioides (Colletotrichum gloeosporioides), sphaerotheca fuliginea (Sphaerotheca fuliginea), botrytis cinerea (Botrytis cinerea), cladosporium fulvidrum (Cladosporium fulvum), sclerotinia sclerotiorum (Sclerotinia sclerororium), verticillium dahliae (Verticillium flaiae), phomopsis vexans (Phomopsis vexans), aschersonia metacarpus (Diabrospirhe vexans), fusarium oxysporum (Fusarium oxysporum), rhizoctonia solani (Rhizoctonia solani), anthrax (C.
5. A product containing a derivative of tris Yang or tris Yang mycin wherein the active ingredient of the product is a derivative of tris Yang or tris Yang, said product having the function of at least one of:
(1) Preventing and treating pathogenic bacteria;
(2) Improving the resistance of plants to pathogenic bacteria;
(3) Inhibiting the pathogenicity of pathogenic bacteria;
(4) Inhibiting the growth of hyphae of pathogenic bacteria;
(5) The fruit preservation is improved;
the molecular structural formula of the tri-Yang mycin is as follows:
6. the product according to claim 5, characterized in that the pathogenic bacteria are Colletotrichum gloeosporioides (Colletotrichum gloeosporioides), sphaerotheca fuliginosum (Sphaerotheca fuliginea), botrytis cinerea (Botrytis cinerea), cladosporium fulvidrum (Cladosporum fulvum), sclerotinia sclerotiorum (Sclerotinia sclerororium), verticillium dahliae (Verticillium flaiae), phomopsis vexans (Phomopsis vexans), aschersonia metacarpus (Diabrospire vexans), fusarium oxysporum (Fusarium oxysporum), rhizoctonia solani (Rhizoctonia solani), anthrax (C.
7. The method for improving the anthracnose resistance of the citrus is characterized by comprising the following specific steps: citrus is treated with an agent comprising a derivative of either tri Yang or tri Yang mycin.
8. The method of claim 7, wherein the agent is a solution containing 10 μ g/mL of tris Yang mycin.
9. The method for improving the freshness preservation of the citrus is characterized by comprising the following specific steps: citrus is treated with an agent comprising a derivative of either tri Yang or tri Yang mycin.
10. The method of claim 9, wherein the reagent is a solution containing 10 μ g/mL of tris Yang mycin and the treating comprises immersing the citrus fruit in the reagent.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS60199864A (en) * | 1984-03-22 | 1985-10-09 | Mitsui Toatsu Chem Inc | Production of n-acylphenylalanine or the like |
| US20070106079A1 (en) * | 2003-12-17 | 2007-05-10 | Dambrin Valery | Novel intermediates for the synthesis of (r)-tamsulosin and of its pharmaceutically acceptable salts and process for their preparation |
| CN101023945A (en) * | 2007-02-16 | 2007-08-29 | 福州大学 | Use of non-periphery substituted phthalocyaniu metal complex |
| WO2021046651A1 (en) * | 2019-09-13 | 2021-03-18 | University Of Prince Edward Island | N-acyl-tyrosine derivatives and uses thereof |
-
2022
- 2022-11-18 CN CN202211463743.3A patent/CN115956564A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS60199864A (en) * | 1984-03-22 | 1985-10-09 | Mitsui Toatsu Chem Inc | Production of n-acylphenylalanine or the like |
| US20070106079A1 (en) * | 2003-12-17 | 2007-05-10 | Dambrin Valery | Novel intermediates for the synthesis of (r)-tamsulosin and of its pharmaceutically acceptable salts and process for their preparation |
| CN101023945A (en) * | 2007-02-16 | 2007-08-29 | 福州大学 | Use of non-periphery substituted phthalocyaniu metal complex |
| WO2021046651A1 (en) * | 2019-09-13 | 2021-03-18 | University Of Prince Edward Island | N-acyl-tyrosine derivatives and uses thereof |
Non-Patent Citations (1)
| Title |
|---|
| ZHAO, LIWEN等: "Facile Synthesis of Chiral Diphosphine-Containing Multiple Dendrimeric Catalysts for Enantioselective Hydrogenation", CHIN. J. CHEM., vol. 30, 31 December 2012 (2012-12-31), pages 2009 - 2015 * |
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