CN114303770A - Method for preventing and controlling pear ring rot by using dopamine - Google Patents
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Abstract
The invention belongs to the technical field of fruit and vegetable disease control, and particularly relates to a method for preventing and controlling pear ring rot by using dopamine, wherein a pear tree body is sprayed by using a dopamine aqueous solution with the concentration of 50-400 mu M in a period of every 15-20 days in a pear fruit expansion period. According to the invention, the activity of phenylalanine ammonia lyase, superoxide dismutase, peroxidase and peroxidase is improved by spraying dopamine, the removal capability of hydrogen peroxide and superoxide anion in pear trees after pathogenic bacteria are infected is increased, the autophagy activity of cells is improved, and the development speed of lesion diameter is delayed, so that the occurrence of ring spot disease is reduced.
Description
Technical Field
The invention particularly relates to a method for preventing and controlling pear ring rot by using dopamine, and belongs to the technical field of fruit tree disease prevention and control.
Background
The ring rot of pear, also called rough bark disease and neoplastic bark disease, is an important disease in pear tree production, China is the biggest pear producing country in the world, and the harvest area, yield, export quantity and variety quantity are in the top of the world. The pear planting range in China is wide, and pear trees are cultivated in other provinces (autonomous regions, city in direct jurisdiction) except the Hainan province, the Hongkong and Australia regions. The ring rot pathogen can attack on pear fruits, branches and leaves, the damage of the branches and the leaves can promote the premature senility of the branches and the leaves can cause the early fall of the leaves, the damage of the fruits can cause a great amount of rotten fruits, and the yield and the quality are seriously influenced. The pear ring rot causes the reduction of the yield of the pears by about 25 percent every year in China, the rotten fruit rate is over 80 percent in severe cases, and in view of the fact that the pear ring rot can seriously affect the production and the sale of pears, the prevention and the treatment of the pear ring rot need to be deeply researched, and more prevention and treatment agents are explored to deal with the pear ring rot.
Most fruit growers suffer from ring spot in the production process of pears by adopting a chemical agent control method, so that not only can the fruits generate resistance to the chemical agent, but also the chemical agent remained on the surfaces of the fruits is unfavorable for human bodies. Therefore, a novel healthy green environment-friendly medicament for preventing and controlling the pear ring rot needs to be explored.
Dopamine is a kind of catecholamine, a biological amine having a structure of a catechol group, and is also present in the human body. At present, the substance is widely applied to animals and human beings, and is less applied to plants.
At present, no report about the effect of dopamine on the biotic stress capability of plants exists.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a method for preventing and controlling the pyricularia piricola by using dopamine.
In order to achieve the purpose, the invention adopts the technical scheme that:
a method for preventing and controlling pear ring rot by using dopamine is characterized in that a pear tree body is sprayed by using a dopamine aqueous solution with the concentration of 50-400 mu M in a period of every 15-20 days in a pear fruit expansion period.
The dopamine of this application can spray to the ripe fruit of pear.
In a preferred embodiment of the present invention, the concentration of the dopamine aqueous solution is 100 μ M.
As the preferable technical scheme, the spraying time is 8-9 in the morning in sunny weather or 8-9 in the afternoon in the mountains, and the spraying is carried out after the weather is slightly cool, so that the rainy weather is avoided.
As a preferred technical scheme of the application, 1% of Tween80 is required to be added when the dopamine aqueous solution is prepared.
A method for preventing and controlling pear ring rot by using dopamine is characterized in that pear fruits, pear leaves and pear branches of treated trees are collected in vitro and inoculated with ring rot manually, and then the lesion diameter is observed periodically.
Specifically, cultivars (such as 'Cuiguan' pear, 'Sucui I' pear, and 'Yulu Xiang' pear) are selected, pear fruits with consistent maturity and size and without mechanical damage and plant diseases and insect pests are selected after being ripened and harvested, the surfaces of the pear fruits are cleaned by sterile water, the middle parts of the fruits are punctured by sterilized needle heads, one hole is punctured every 120 degrees, ring vein pathogenic bacteria are inoculated, and then the pear fruits are cultured in a constant-temperature incubator at 25 ℃ for regularly observing the diameters of disease spots.
More specifically, the lesion diameter is observed after 5 to 10 days, preferably 6 days.
Specifically, pricking the upper and lower positions of the left and right sides of a pear leaf, inoculating ring rot pathogenic bacteria, culturing at 25 ℃, and regularly observing the diameter of a lesion spot. Specifically, a puncher with the diameter of 0.5 cm is used for punching the pear branches after spraying treatment, the epidermis is broken, and after ring-line pathogenic bacteria are inoculated, the inoculated part is covered by a preservative film to prevent fungus cakes from falling off; spraying clear water at two ends of the branch, winding a preservative film to preserve water, and regularly observing the diameter of the lesion spots.
More specifically, the lesion diameter is observed after 5 to 10 days, preferably 8 days.
Specifically, the depth of the puncture hole on the surface of the fruit is 5 mm, and the diameter of the puncture hole is 2 mm; the diameter of the blade hole is 2 mm; the diameter of the branch hole is 5 mm.
Wherein, the temperature of 25 ℃ is the optimum temperature for the activity of pathogenic bacteria of the ring spot, and the diameter of the lesion spot is easy to be observed at the temperature.
Advantageous effects
Compared with the prior art, the method for preventing and controlling the pear ring rot by using the dopamine has the following beneficial effects:
(1) the method creatively discovers that the resistance of pears to the ring spot can be improved by spraying dopamine solution with the concentration of 50-400 mu M in a 15-20 day period in a fruit expanding period, the removal capability of hydrogen peroxide and superoxide anions in pear trees after pathogenic bacteria are infected is improved by improving the activities of phenylalanine ammonia lyase, superoxide dismutase, peroxidase and peroxidase, the autophagy activity of cells is improved, the development speed of the diameter of the lesion spot is delayed, and the ring spot is reduced.
(2) Dopamine is a water-soluble substance, is convenient for fruit growers to use, and can be directly dissolved in tap water or river water, well water and other irrigation water.
(3) The cost of the dopamine is low, the dopamine concentration is about 5g per mu according to the test, and the cost is about 32.7 yuan per mu in terms of market cost. According to different pear production periods in Nanjing, the early-maturing pears are sprayed for 4-6 times in the expansion period, the medium-maturing pears are sprayed for 6-8 times, and the late-maturing pears are sprayed for 8-10 times. The cost can be effectively saved, the use of pesticides is reduced, dopamine is a substance which can be synthesized by a human body, no harmful residue is left when fruits are sprayed, no harm is caused to the human body, and the fruit quality can be improved to a certain extent. Provides a new idea for producing healthy and green pear fruits.
Drawings
FIG. 1 shows the inhibitory effect of dopamine treatment at different concentrations on Phyllostachys virens;
FIG. 2 shows the effect of dopamine treatment on the development of lesion diameter of ring rot disease of Cuiguan pear fruit;
FIG. 3 shows the effect of dopamine treatment on the development of lesion diameter of a green crown pear branch ring spot;
FIG. 4 is a graph of the effect of dopamine treatment on lesion diameter development of the ring spot disease of Sucui I pear fruit;
FIG. 5 is a graph of the effect of dopamine treatment on lesion diameter development of 'Sucui I' pear branch ring rot;
FIG. 6 shows the effect of dopamine treatment on the development of lesion diameter of the ring spot disease of the 'Yulu Xiang' pear fruit;
FIG. 7 shows the effect of dopamine treatment on lesion diameter development of a bouquet on Verticillium disease;
FIG. 8 shows the effect of dopamine treatment on the development of the lesion diameter of leaf stripe disease of Dangshan pear;
FIG. 9 shows the effect of dopamine treatment on the SOD enzyme activity of Dangshan pear leaf inoculated ring spot for 6 days;
FIG. 10 shows the effect of dopamine treatment on the POD enzyme activity of Dangshan pear leaf inoculation for 6 days;
FIG. 11 shows the effect of dopamine treatment on the CAT enzyme activity of Dangshan pear leaf inoculated ring spot for 6 days;
FIG. 12 shows the effect of dopamine treatment on the SOD enzyme activity of Dangshan pear leaf inoculated ring spot for 6 days;
FIG. 13 shows the effect of dopamine treatment on the OFR content of Dangshan pear leaf inoculated ring spot for 6 days;
FIG. 14 shows that 6 days H of the leaf inoculation of Dangshan pear2O2Influence of the content;
FIG. 15 shows the effect of dopamine treatment on autophagy activity of Dangshan pear.
Detailed description of the invention
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.
The concrete implementation places are as follows: jinling crisp pear garden in Jiangning region lake of Jiangsu province, Nanjing city
The detection method comprises the following steps:
1. after the pear fruits are ripe and harvested, the pear fruits with the same maturity and size and without mechanical damage and plant diseases and insect pests are selected, the surfaces of the pear fruits are cleaned by sterile water, the middle parts of the pear fruits are punctured by sterilized needles, one hole is punctured every 120 degrees, ring rot pathogenic bacteria are inoculated, and the diameters of disease spots are measured after the pear fruits are cultured for 6 days at the temperature of 25 ℃.
2. Pricking the left side and the right side of the pear leaf in the upper and lower directions, inoculating ring rot pathogenic bacteria, culturing at 25 deg.C, and measuring the diameter of lesion spot in 0, 2, 4, 6, and 8 days.
3. And (3) punching the pear branches subjected to spraying treatment by using a puncher with the diameter of 0.5 cm, breaking the epidermis, inoculating ring rot pathogenic bacteria, and covering the inoculation position with a preservative film to prevent the fungus cakes from falling off. Spraying clear water on two ends of the branch, winding a preservative film for water retention, culturing for 8 days in a constant-temperature incubator at 25 ℃, and observing the diameter of the lesion spots.
4. Respectively measuring Phenylalanine Ammonia Lyase (PAL), superoxide dismutase (SOD), peroxidase (CAT), Peroxidase (POD) activity and hydrogen peroxide (H) in treated pear leaves inoculated with ring rot pathogenic bacteria2O2) Content and superoxide anion (OFR) content.
5. The enzyme activity, hydrogen peroxide and superoxide anion content are detected by using a kit produced by Suzhou Keming biology.
6. Detecting the number of autophagosomes: at 6d, pieces of lamina at the interface of the lamina diseased keys were excised and fixed at 4 ℃ for 12h in 0.2M PBS buffer (pH 7.4) containing 2.5% glutaraldehyde. After washing with 0.2M PBS (pH 7.2) buffer, the samples were fixed in 1% (v/v) osmium tetroxide for 2.5h at room temperature. A gradient ethanol solution (30-100%; v/v) was used for dehydration and embedding in Epon 812. Ultrathin sections (70nm) were cut out using a microtome (Leica ULTRACUT, Germany) and placed on Formvar and autophagosomes were observed under a JEOL-1230 Transmission Electron microscope (Hitachi, Japan) at a voltage of 80 kV. The number of autophagosomes per cell was counted, and at least 10 cells per replicate were counted.
Example 1 prevention and control of Pear stripe disease with dopamine at different concentrations
Setting four dopamine aqueous solutions with concentrations of 50, 100, 200 and 400 mu M, spraying 30 dopamine aqueous solutions with four concentrations on the leaves of the Dangshan pear which is harvested by using clear water as a control, and inoculating ring rot germs after 24 hours of treatment. The lesion diameter was counted every two days.
The test results of indoor spraying of dopamine with different concentrations are shown in fig. 1, the dopamine with four concentrations of 50, 100, 200 and 400 mu M can inhibit growth of the pear ring rot, and the diameters of lesions of the pear ring rot are respectively reduced by 41.6%, 71.6%, 54% and 62.3%. Among them, 100 μ M dopamine was the best treated, so subsequent experiments were performed with this concentration.
Example 2
3 plants of 'Cuiguan', 'Sucui No. I' and 'Yulu Xiang' with the same tree vigor, tree age and load capacity are selected for testing. Spraying 100 mu M dopamine solution every 15-20 days in the fruit expansion period, and adding 1% Tween 80. 'Sucui No. one' spraying time is 4 times (5 months and 1 day, 5 months and 21 days, 6 months and 11 days and 7 months and 1 day in 2020), respectively, 'Cuiguan' spraying time is 5 times (5 months and 1 day, 5 months and 21 days, 6 months and 11 days, 7 months and 1 day and 7 months and 21 days in 2020), and 'Yulu Xiang' spraying time is 8 times (5 months and 1 day, 5 months and 21 days, 6 months and 11 days, 7 months and 1 day, 7 months and 21 days, 8 months and 10 days, 9 months and 1 day and 9 months and 20 days in 2020). The spraying time is 8-9 am in sunny weather or 4-5 pm in the afternoon, and clear water is sprayed as a control test.
Selecting pear fruits with consistent maturity and size and without mechanical damage and plant diseases and insect pests, cleaning the surface of the pear fruits by using sterile water, puncturing the middle parts of the pear fruits by using a sterilized needle, puncturing one hole every 120 degrees, inoculating ring rot pathogenic bacteria, culturing for 6 days at 25 ℃, and measuring the diameter of a lesion. Lesion diameter test results referring to figures 2-7, as shown, the tests found that the mean lesion diameter was significantly reduced after 6 days of inoculation with ring rot pathogen for 'peri' pear fruit, 'emerald' pear fruit and 'yulu fragrant' pear fruit after 100 μ M treatment with dopamine compared to CK treatment; the reduction is 19.2 percent, 16.1 percent and 34.4 percent respectively.
And (3) punching the pear branches subjected to spraying treatment by using a puncher with the diameter of 0.5 cm, breaking the epidermis, inoculating ring rot pathogenic bacteria, and covering the inoculation position with a preservative film to prevent the fungus cakes from falling off. Spraying clear water on two ends of the branch, winding a preservative film for water retention, culturing for 8 days at 25 ℃, and observing the diameter of the lesion spots. After being treated by 100 mu M of dopamine, the 'Sucui I' pear branches, the 'Cucuiguan' pear branches and the 'Yulu Xiang' pear branches are inoculated with ring grain pathogenic bacteria for 6 days, and then the average lesion diameter is obviously reduced compared with CK treatment. Respectively reduced by 18.3%, 15.4% and 34.1%.
Example 3
3 Dangshan pears with the same tree vigor, tree age and load capacity are selected for testing.
Pricking the left side and the right side of the pear leaf in the upper and lower directions, inoculating ring rot pathogenic bacteria, culturing at 25 deg.C, and measuring the diameter of lesion spot in 0, 2, 4, 6, and 8 days. After 100 mu M dopamine treatment, the average lesion diameter of Dangshan pear leaves after inoculation of ring spot pathogenic bacteria is significantly lower than that of CK treatment in 2, 4, 6 and 8 days, and is respectively reduced by 62.2%, 37.3%, 29.1% and 26.1%, as shown in the lesion diameter of figure 8.
Respectively measuring Phenylalanine Ammonia Lyase (PAL), superoxide dismutase (SOD), peroxidase (CAT), Peroxidase (POD) activity and hydrogen peroxide (H) in treated pear leaves inoculated with ring rot pathogenic bacteria2O2) Content and superoxide anion (OFR) content. Referring to fig. 9-13, as shown in the figure, after 6 days of inoculating the physalospora bacteria to the Dangshan pear leaves, the activities of Phenylalanine Ammonia Lyase (PAL), superoxide dismutase (SOD), peroxidase (CAT) and Peroxidase (POD) are significantly improved by 10.2%, 30%, 18.2% and 13.7% respectively, compared with CK treatment. Hydrogen peroxide (H)2O2) And superoxide anion (OFR) content was significantly reduced compared to CK treatment, by 10% and 14.8%, respectively.
Referring to fig. 14, after 100 μ M of dopamine treatment, the autophagy activity of the Dangshan pear leaves was significantly improved 6 days after inoculation of the pathogenic ring spot, compared to the CK treatment. The number of autophagosomes of the Dangshan pear inoculated with the ring spot germs after treatment is increased by 377 percent compared with that of the Dangshan pear inoculated with the ring spot germs after the treatment, and the autophagy activity of the Dangshan pear is obviously increased.
In summary, dopamine is produced by increasing Phenylalanine Ammonia Lyase (PAL), superoxideThe activities of dismutase (SOD), peroxidase (CAT) and Peroxidase (POD) enhance the hydrogen peroxide (H) generated in the process of infecting pear with ring rot pathogen2O2) And superoxide anion (OFR) scavenging ability; the dopamine can also improve the autophagy activity of pears, enhance the autophagy capability of damaged cells, improve the resistance of pears to the ring rot fungi and delay the infection speed of the ring rot fungi.
The protection of the present invention is not limited to the above embodiments. Variations and advantages that may occur to those skilled in the art may be incorporated into the invention without departing from the spirit and scope of the inventive concept and the scope of the appended claims is intended to be protected.
Claims (10)
1. A method for preventing and controlling pear ring rot by using dopamine is characterized in that a pear tree body is sprayed by using a dopamine aqueous solution with the concentration of 50-400 mu M in a period of every 15-20 days in a pear fruit expansion period.
2. The method for preventing and controlling pyricularia piricola by using dopamine, according to claim 1, wherein the concentration of the dopamine aqueous solution is 100 μ M.
3. The method for preventing and controlling the pyricularia pyricularis by using the dopamine according to claim 1 or 2, characterized in that spraying is performed when the time is selected to be 8-9 o' clock in the morning of sunny weather or after the weather is slightly shady and cool in the afternoon and the sun, and the rainy weather is avoided.
4. The method for preventing and controlling the pyricularia pyricularis by using the dopamine according to claim 1, wherein 1% of Tween80 is added when the dopamine aqueous solution is prepared.
5. A detection method for preventing and controlling pear ring rot by using dopamine is characterized in that pear fruits, pear leaves and pear branches of treated trees are collected in vitro and inoculated with ring rot manually, and then the lesion diameter is observed periodically.
6. The method for detecting pear streak disease prevention and control by using dopamine according to claim 5, characterized in that, a cultivar is selected, pear fruits with consistent maturity and size and without mechanical damage and plant diseases and insect pests are selected after being ripe and harvested, the surface of the pear fruits is cleaned by sterile water, the middle parts of the pear fruits are punctured by sterilized needles, one hole is punctured every 120 degrees, ring streak pathogenic bacteria are inoculated, then the pear fruits are cultured in an environment of 25 ℃, and the diameters of disease spots are regularly observed.
7. The detection method for preventing and controlling pear leaf streak disease by using dopamine as claimed in claim 5, wherein pricking holes are performed on the left side, the right side, the upper side and the lower side of a pear leaf, ring rot pathogenic bacteria are inoculated, cultivation is performed in an environment of 25 ℃, and the diameter of a lesion spot is regularly observed.
8. The detection method for preventing and controlling pear leaf spot by using dopamine according to claim 5, characterized in that a puncher with a diameter of 0.5 cm is used for punching pear branches after spraying treatment, the epidermis is broken, and after ring spot pathogenic bacteria are inoculated, a preservative film is used for covering the inoculated part to prevent fungus cakes from falling off; spraying clear water at two ends of the branches, winding a preservative film to preserve water, and regularly observing the diameters of the disease spots; preferably, the diameter of the branch holes is 5 mm.
9. The detection method for preventing and controlling pear leaf spot by using dopamine as claimed in claim 6, wherein the depth of the pricked hole on the fruit surface is 5 mm, and the diameter of the hole is 2 mm.
10. The detection method for preventing and controlling pear leaf spot by using dopamine as claimed in claim 7, wherein the diameter of the leaf hole is 2 mm; the diameter of the branch hole is 5 mm.
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| CN103503737A (en) * | 2013-10-09 | 2014-01-15 | 中国农业科学院郑州果树研究所 | Method for identifying resistance of cytosporacarphosperma |
| CN111471741A (en) * | 2020-06-10 | 2020-07-31 | 南京农业大学 | Method for rapidly identifying ring spot resistance of pear trees |
| CN111656996A (en) * | 2020-07-02 | 2020-09-15 | 凯里学院 | Method for preventing and controlling golden autumn pear ring rot by using methyl jasmonate |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103503737A (en) * | 2013-10-09 | 2014-01-15 | 中国农业科学院郑州果树研究所 | Method for identifying resistance of cytosporacarphosperma |
| CN111471741A (en) * | 2020-06-10 | 2020-07-31 | 南京农业大学 | Method for rapidly identifying ring spot resistance of pear trees |
| CN111656996A (en) * | 2020-07-02 | 2020-09-15 | 凯里学院 | Method for preventing and controlling golden autumn pear ring rot by using methyl jasmonate |
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