CN115886028B - Compound medicament and application thereof in preventing and treating bacterial spot of tomatoes - Google Patents
Compound medicament and application thereof in preventing and treating bacterial spot of tomatoes Download PDFInfo
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- CN115886028B CN115886028B CN202211507345.7A CN202211507345A CN115886028B CN 115886028 B CN115886028 B CN 115886028B CN 202211507345 A CN202211507345 A CN 202211507345A CN 115886028 B CN115886028 B CN 115886028B
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Classifications
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The application discloses a compound medicament and application thereof in preventing and treating tomato bacterial spot disease, and belongs to the technical field of biology. The technical field of green prevention and treatment of tomato bacterial spot disease. The research of the application finds that the potassium silicate and linalool have inhibition effect on tomato bacterial spot disease pathogenic bacteria-pseudomonas syringae tomato pathogenic variety (Pseudomonas syringae pv.total, pst); the compound preparation of the two can not only effectively inhibit the bacterial leaf spot of tomatoes, but also promote the root system activity of plants to a certain extent to promote the plant growth. The application performs root irrigation treatment on tomatoes by compounding the medicament, can play an effective role in preventing and treating bacterial spot diseases of tomatoes, and has important significance in preventing and treating bacterial spot diseases of tomatoes. Compared with the traditional chemical agents, the compound agent of the potassium silicate and the linalool has the advantages of equivalent prevention effect, environmental friendliness, no pesticide residue and stable prevention effect.
Description
Technical Field
The application relates to the technical field of green control of bacterial spot diseases of tomatoes, in particular to a compound medicament and application thereof in controlling bacterial spot diseases of tomatoes.
Background
Tomato bacterial leaf spot (Tomato bacterial leaf spot), also known as tomato bacterial leaf spot, is one of the important diseases that jeopardize tomato production worldwide. In recent years, along with the continuous expansion of the cultivation area of the facility tomatoes, the occurrence degree of the disease is in a continuous rising trend, and the healthy development of the tomato industry is seriously affected. Pseudomonas syringae tomato pathogenic variety (Pseudomonas syringae pv.cyto, pst) is an aerobic, saprophytic gram negative bacterium, the thallus is in a short rod shape, the size is (0.5-1.0) mu m x (1.5-5.0) mu m, and the tomato pathogenic variety has 1 to several polar flagella, no capsule and no spore. The bacteria form milky circular colonies after being cultured on KB culture medium for 48 hours, the diameters of the colonies are 2-3 mm, the whole edges are opaque, the surfaces are smooth and sticky, and yellow-green fluorescence is observed under an ultraviolet lamp. The germ can overwinter tomato plants, seeds, disease residues, soil and field weeds, and becomes an initial infection source in the next year. Therefore, the tomatoes in winter and spring protection areas are often seriously damaged. Bacterial spot disease of tomato caused by pseudomonas syringae tomato pathogenic variety (Pseudomonas syringae pv.mat, pst) tomato pathogenic variety (Pseudomonas syringae pv.mat, pst) is mainly harmful to tomato leaves, can also infect stems and fruits, and can be developed in both seedling stage and adult stage of tomato. The leaf blade in seedling stage produces water stain round dot-shaped disease spots in the early stage of disease onset, the disease spots are dark brown after being expanded, and the periphery of the disease spots turn yellow and green; the disease spots become brown or black in middle and late stages of the disease, and the disease spots can continuously grow a plurality of disease spots along the veins to cause the malformation of the leaves. When the leaf is infected, the lower mature aging leaf is infected before spreading to the upper part of the plant.
The potassium silicate has wide application, one of the potassium silicate can be used as a fertilizer, and the potassium silicate fertilizer can improve soil, correct acidity of the soil, promote organic fertilizer to be divided and inhibit soil pathogens; plant extracts and essential oils are often broad-spectrum bactericidal as a class of natural organic compounds, wherein linalool is an organic compound rich in plant materials such as pricklyash seeds. At present, no research report on application of potassium silicate and linalool to tomato bacterial spot disease is seen.
Disclosure of Invention
Aiming at the prior art, the application aims to provide a compound medicament and application thereof in preventing and treating bacterial spot of tomatoes.
In order to achieve the above purpose, the application adopts the following technical scheme:
in a first aspect of the present application, there is provided a compound medicament comprising potassium silicate and linalool as active ingredients.
Preferably, the compound medicament is prepared by compounding 0.8mL/L linalool solution and 1.75g/L potassium silicate solution according to any volume ratio as follows:
(1) The volume ratio of the potassium silicate solution to the linalool solution is 1:2;
(2) The volume ratio of the potassium silicate solution to the linalool solution is 1:1;
(3) The volume ratio of the potassium silicate solution to the linalool solution is 2:1.
In a second aspect of the application there is provided the use of a combination medicament as described above in the preparation of a fungicide for the pathogenic variety of Pseudomonas syringae tomato (Pseudomonas syringae pv. Mat, pst).
In a third aspect of the present application, there is provided the use of the above-described compound medicament in (1) or (2) as follows:
(1) Preventing and treating bacterial spot disease of tomato;
(2) Preparing the product for preventing and treating tomato bacterial spot.
In the above application, the pathogenic bacteria of bacterial spot disease of tomato is Pseudomonas syringae tomato pathogenic variety (Pseudomonas syringae pv.
In a fourth aspect of the application, there is provided a method of controlling bacterial spot in tomatoes comprising the steps of:
after the tomato is planted, the compound preparation is adopted to irrigate the root of the tomato plant.
Preferably, the root irrigation treatment is performed after the 5-6 leaf stage of the tomato seedling age.
Preferably, the dosage of the compound preparation is 30ml per tomato plant; once every 3 days, 5 consecutive administrations.
The application has the beneficial effects that:
(1) The application is found in the first study: the potassium silicate and linalool have obvious inhibition effect on pseudomonas syringae tomato pathogenic varieties (Pseudomonas syringae pv.total, pst), and can effectively reduce the incidence rate of tomato bacterial spot disease through root irrigation treatment, thus having great significance for preventing and treating the tomato bacterial spot disease.
(2) The tomato root irrigation treatment by potassium silicate and linalool can effectively prevent and treat pseudomonas syringae tomato pathogenic varieties (Pseudomonas syringae pv. Total, pst), and has important significance for preventing and treating bacterial spot diseases of tomatoes. And compared with the traditional chemical agents, the potassium silicate and linalool have equivalent control effect, are environment-friendly, have no pesticide residue and have stable control effect.
Drawings
Fig. 1: effect of different agent treatments on root system activity of tomato plants.
Fig. 2: effects of different agent treatments on chlorophyll content and carotenoid content of tomato plants.
Fig. 3: effects of different agent treatments on tomato plants Φpsii, fv/Fm.
Fig. 4: grading the disease states of the tomato bacterial spot diseases.
Fig. 5: the different treatments have preventive effects on bacterial spot of tomato.
Detailed Description
It should be noted that the following detailed description is illustrative and is intended to provide further explanation of the application. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
Bacterial spot disease of tomatoes is a very harmful tomato soil-borne disease caused by pseudomonas syringae tomato pathogenicity (Pseudomonas syringae pv. Mat, pst), which causes a great loss in tomato production in our country. Pathogenic bacteria can overwinter in seeds, plant disease and residues and soil. Sowing the seed with bacteria, curing the seedlings, curing the disease after the field planting of the disease seedlings, spreading the disease in a large area through rain back splashing, agricultural operation, irrigation water to spread the disease and the like; the flood irrigation, cold and wet environment and low temperature and rainy environment are also beneficial to disease occurrence. At present, the product for preventing and treating the bacterial spot disease of tomatoes is mainly chemical agent and is single. Therefore, there is a need to develop green and environment-friendly products for preventing and treating bacterial spot of tomatoes.
Linalool is a monoterpene volatile substance, and has the chemical structural formula as follows:
it is present in the roots, stems, leaves, flowers, fruits of a wide variety of plants, the method has the characteristics of green, safety, high efficiency and the like, and is environment-friendly; the antibacterial effect is good, and resistance is not easy to generate; the problems of pesticide residue, environmental pollution and the like are not caused, and the environment-friendly development concept is met.
The potassium silicate has wide application, and one of the potassium silicate can be used as fertilizer. Firstly, the potassium silicate fertilizer is a nutrient element necessary for a plurality of plants, and secondly, the potassium silicate fertilizer is a nutrient element fertilizer with high nutrition, has good effects on improving the pH value of soil and promoting the differentiation of soil nutrients, and can ensure the yield and income of crops to a great extent by using the potassium silicate fertilizer. In addition, the potassium silicate can also improve the oxidation capacity of the root system and the respiration rate of the root system, enhance the absorption capacity of the root system to nutrients in water, and increase the ATP content in leaves and the root system so that the nutrients are directly absorbed and converted and transported to fruits).
It is present in the roots, stems, leaves, flowers, fruits of a wide variety of plants, the method has the characteristics of green, safety, high efficiency and the like, and is environment-friendly; the antibacterial effect is good, and resistance is not easy to generate; the problems of pesticide residue, environmental pollution and the like are not caused, and the environment-friendly development concept is met.
Experiments show that the potassium silicate and linalool have obvious inhibition effect on pseudomonas syringae tomato pathogenic varieties (Pseudomonas syringae pv.total, pst), and can effectively reduce the incidence of tomato bacterial spot by root irrigation treatment, thus having great significance for preventing and treating tomato bacterial spot. The present application has been made in view of the above.
In order to enable those skilled in the art to more clearly understand the technical scheme of the present application, the technical scheme of the present application will be described in detail with reference to specific embodiments.
The test materials used in the examples of the present application are all conventional in the art and are commercially available. The experimental procedure, without specifying the detailed conditions, was carried out according to the conventional experimental procedure or according to the operating instructions recommended by the suppliers. Wherein: linalool was purchased from alas Ding Shiji (Shanghai) limited); potassium silicate is purchased from national pharmaceutical group chemical reagent limited; pseudomonas syringae tomato pathogenic variety (Pseudomonas syringae pv.mat, pst), described in non-patent literature "function of tomato S1βCA3 in defense against Pseudomonas syringae tomato pathogenic variety: 2740-2751, "20 years from the date of application, the public is available from the applicant for use in repeating the present application.
Example 1: effect of potassium silicate and linalool on the growth of pseudomonas syringae tomato pathogenic variety (Pseudomonas syringae pv.
Activating strains: the bacterial liquid of pathogenic variants (Pseudomonas syringae pv.mat, pst) of Pseudomonas syringae tomato stored at low temperature is coated on a KB solid culture medium plate for culturing for 24 hours at 28 ℃, single colony is selected and cultured for 8 hours at 28 ℃ in the KB liquid culture medium plate.
KB solid medium formula: bacto TM Proteose peptone 29g/L, K 2 HPO 4 ·3H 2 O 1.96g/L，Glycerol 8mL/L，MgSO 4 ·7H 2 O 1.52g/L，Agar 15g/L，Rifampin 1mL/L。
KB liquid medium formula: bacto TM Proteose peptone 29g/L, K 2 HPO 4 ·3H 2 O 1.96g/L，Glycerol 8mL/L，MgSO 4 ·7H 2 O 1.52g/L，Rifampin 1mL/L。
Respectively adding a certain amount of potassium silicate into a liquid KB culture medium to prepare culture mediums with the final concentration of potassium silicate of 0mL/L, 0.4g/L, 0.8g/L, 1.2g/L, 1.6g/L and 2 g/L; and respectively adding a certain amount of linalool into the liquid KB culture medium to prepare culture mediums with final linalool concentrations of 0mL/L, 0.4mL/L, 0.8mL/L, 1.2mL/L, 1.6mL/L and 2 mL/L. The activated bacterial solution was centrifuged and resuspended in sterile water to a cell density OD600 of 0.5.
The treatment group comprises the following steps: the medicine-containing culture medium (volume ratio) is mixed in the ratio of 1:100. Liquid KB medium without adding medicament is used as a Control (CK), and shake culture is carried out for 12 hours at 28 ℃ and 180 r/min.
Bacteria and method for producing sameGrowth relative bacteriostasis rate, bacteriostasis rate value, virulence regression equation and EC 50 The value calculation formula is as follows:
(1)
(2) Antibacterial several values (y) =NORMINV (relative antibacterial ratio, 5, 1)
(3) Virulence regression equation: y=ax+b, and when y=5, the obtained x value is the logarithm of the EC50 value
X axis: log, log_10x of agent concentration
Y axis: inhibition rate value, NORMSINV () +5
R value: CORREL (x, y)
Cut-off a value: INTERCEPT (x, y)
Slope b value: SLOPE (x, y)
(4)EC50＝POWER(10，x)
Relative inhibition rate, inhibition rate value and half maximum Effect Concentration (EC) of potassium silicate with each concentration on bacterial growth of pseudomonas syringae tomato pathogenic variety (Pseudomonas syringae pv.total, pst) 50 ) See table 1.
Table 1: relative inhibition of P.syringae tomato pathogenic variety (Pseudomonas syringae pv. Mat, pst) butyl by potassium silicate, virulence regression equation and EC50 value
The results show that potassium silicate has a remarkable inhibition effect on the growth of the filament of the pathogenic variety (Pseudomonas syringae pv. Cyto, pst) of Pseudomonas syringae tomato.
Relative inhibition rate, inhibition rate value and half maximum Effect Concentration (EC) of linalool at each concentration on bacterial growth of pseudomonas syringae tomato pathogenic variety (Pseudomonas syringae pv.total, pst) 50 ) See table 2.
Table 2: relative inhibition of linalool against P.syringae tomato pathogenic variety (Pseudomonas syringae pv. Mat, pst), virulence regression equation and EC 50 Value of
The results show that linalool has a remarkable inhibition effect on the growth of the filaments of the pathogenic variety of Pseudomonas syringae tomato (Pseudomonas syringae pv. Total, pst).
Example 2: optimal formulation of both potassium silicate and linalool screening for effects on the growth of pseudomonas syringae tomato pathogenic variety (Pseudomonas syringae pv. Toolo, pst).
According to the results of example 1, potassium silicate (1.75 g/L) and linalool (0.8 ml/L) were mixed in a volume ratio (1:4), (1:2), (1:1), (2:1), (4:1) to prepare a compound 1, a compound 2, a compound 3, a compound 4, a compound 5, and EC of the different compound agents were measured 50 Values. And (3) calculating an SR value by using the formulas (1) and (2), wherein the SR value is synergistic when the SR value is more than or equal to 1.5, antagonistic when the SR value is less than 0.5, and additive when the SR value is between 0.5 and 1.5.
(1)
(2)
A and B are compound medicaments; a, b is the proportion of the medicament in the compounding process; EC (EC) 50 (Exp) is the concentration in theoretical inhibition, EC 50 (Obs) is the concentration in the actual measured inhibition.
Table 3: different concentration design of compound medicament of each group
Table 4: each compound medicament is used for treating pseudomonas syringaeVirulence regression equation of tomato pathogenic variety (Pseudomonas syringae pv. Mat, pst), concentration EC in actual measurement inhibition 50 (Obs), theoretical inhibition of intermediate concentration EC 50 (Exp) and synergistic ratio
SR
The test results show that when the compound agents are compounded according to the volume ratio of 1:4, 1:2, 1:1, 2:1 and 4:1, the compound agents have synergism when potassium silicate and linalool are compounded according to the volume ratio of 1:2, 1:1 and 2:1, and the SR value is between 1.66 and 2.612, wherein the volume ratio of the potassium silicate to linalool is 1: the synergy is best in the 1 time, and the SR value is 2.612; the added effect is achieved when the volume ratio of potassium silicate to linalool is 4:1.
Example 3: safety test of composite medicament on tomato plants
4 tomato plants are set for treatment, and tomato plants in 5-6 leaf stages are taken for testing.
Test grouping:
CK: treating with clear water;
t1: root irrigation treatment of potassium silicate (1.75 g/L);
t2: root irrigation treatment with linalool (0.8 ml/L);
t3: root irrigation treatment is carried out by a compound medicament, and the compound medicament is formed by compounding 1.75g/L potassium silicate and 0.8ml/L linalool according to the volume ratio of 1:1.
Each treatment group is treated once every three days, and is continuously treated for 5 times; root irrigation treatment is carried out according to the dosage of 30mL; root vigor of each group of plants was measured (measurement methods were referred to as Abdul Rehman Khan, zhihui Cheng, bushara Ghazanfar, muhammad Azam Khan & Zhu yongxing. Actyl salicylic acid and-epibrassinolide enhance root activity and improve root morphological features in tomato plants under heat stress, acta Agriculturae Scandinavica, section B-oil & Plant Science, 2014:4, 304-311), chlorophyll content (measurement methods were referred to as Ding, s, zhang, y, hu, z, huang, x, zhang, B, lu, q, wen, x, wang, y, lu, c, 2019.mTERF5 acts as atranscriptional pausing factor to positively regulate transcription of chloroplast psbEFLJ.Mol.Plant 12,1259-1277), photosynthesis (measurement methods were referred to as Gu Wenfei, wei Xiaoqiong, zhang Qiuying, li Linyu, wang Ying, li Jinying, wu Lin. Influence of saline-alkali treatment on photosynthesis characteristics of citrus, chlorophyll, agricultural Science, 2022,50 (07): 152-158), etc. see fig. 1-3.
The results show that the application of the medicaments of each treatment group can improve the root system activity of plants to a certain extent, enhance the photosynthesis of the plants and promote the growth of the plants.
Example 4: prevention effect of composite medicament on bacterial spot disease of tomatoes
4 tomato plants are set for treatment, and tomato plants in 5-6 leaf stages are taken for testing.
Test grouping:
CK: treating with clear water and inoculating;
t1: potassium silicate (1.75 g/L) treatment + inoculation;
t2: linalool (0.8 ml/L) treatment + inoculation;
t3: treatment and inoculation of a compound medicament, wherein the compound medicament is prepared from 1.75g/L potassium silicate and 0.8mL/L linalool according to a volume ratio of 1:1.
Note that: the root irrigation treatment method (root irrigation is carried out once every 3 days and 5 times are carried out continuously) is adopted for each treatment, and the application is carried out according to the usage amount of 30mL of each plant.
The pathogenic bacteria Pseudomonas syringae tomato pathogenic variety (Pseudomonas syringae pv. Mat, pst) was inoculated in KB solid medium and cultured at 28 ℃. Single colonies were picked in 2KB liquid medium, cultured at 28℃for 8-12 h at 200r/min to OD600 = 0.8-1.0. Centrifugation was performed at 4℃for 5min at 4,000Xg, the supernatant was discarded, washed twice with sterile water and resuspended to a concentration of OD600 = 0.08-0.1 and silwet L-77 surfactant was added to a final concentration of 0.01%. And after root irrigation is carried out for 5 times, she Penfa is adopted for pathogen inoculation within 24 hours after the last root irrigation, and pathogen suspension is uniformly sprayed on the surfaces of the blades. The morbidity condition is counted on the 5 th day, and the morbidity rate and the disease index are calculated by the following calculation method:
(1) Incidence = number of diseased seedlings/total number of seedlings x 100%
(2) Disease index= (Σnumber of disease stages×disease classification representative value)/highest disease stage number×total number of investigation×100
(3) Relative control effect = (control group disease index-treatment group disease index)/control group disease index
The photographs were taken after the statistics were completed, and specific phenotypes are shown in fig. 4 and 5.
Grading the disease states of tomato bacterial spot diseases:
level 0: the plants are normal, and no obvious disease spots exist;
stage 1: a small part of the plant leaves yellow, and the proportion of the disease spot area to the leaf area is less than or equal to 25%;
2 stages: most of the leaves of the plants turn yellow, and the edges of a small part of the leaves turn burnt, and the proportion of the area of the disease spots which is more than 25 percent and accounts for less than or equal to 50 percent of the area of the leaves;
3 stages: the whole plant is yellowing and atrophy, most of leaf edges are burnt, and the proportion of the area of the disease spots which is more than 50% and less than or equal to 75% of the leaf area is reduced;
4 stages: the plant wilts and dies, and the proportion of the area of the plant wilts and the area of the plant spots is 75 percent to less than or equal to 100 percent.
Table 6: the incidence rate, incidence index and relative prevention and treatment effect of the bacterial spot disease of the tomatoes after each group of treatment.
The result shows that the incidence rate and the incidence index of the bacterial spot disease of the tomato after the treatment by the composite medicament are obviously smaller than those of a control group, the relative control effect is obviously improved compared with that of the single medicament treatment, and the resistance of the tomato plant to pseudomonas syringae is effectively enhanced.
The above description is only of the preferred embodiments of the present application and is not intended to limit the present application, but various modifications and variations can be made to the present application by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the protection scope of the present application.
Claims (7)
1. The compound medicament is characterized by being prepared by compounding 0.8mL/L linalool solution and 1.75g/L potassium silicate solution according to any volume ratio as follows:
(1) The volume ratio of the potassium silicate solution to the linalool solution is 1:2;
(2) The volume ratio of the potassium silicate solution to the linalool solution is 1:1;
(3) The volume ratio of the potassium silicate solution to the linalool solution is 2:1.
2. The method for preparing pseudomonas syringae tomato pathogenic variety using the compound preparation of claim 1Pseudomonas syringae pv.tomato，Pst) Is used as bactericide.
3. The use of the compounded medicament of claim 1 in (1) or (2) as follows:
(1) Preventing and treating bacterial spot disease of tomato;
(2) Preparing the product for preventing and treating tomato bacterial spot.
4. The use according to claim 3, wherein the pathogenic bacteria of bacterial spot disease of tomato is pseudomonas syringae tomato pathogenic variety @Pseudomonas syringae pv.tomato，Pst)。
5. A method for controlling bacterial spot disease in tomatoes, comprising the steps of:
after tomato field planting, the compound preparation of claim 1 is used for root irrigation treatment of tomato plants.
6. The method according to claim 5, wherein the root irrigation treatment is performed after a period of 5-6 leaves of tomato seedling age.
7. The method of claim 5, wherein the amount of the compounded agent used is 30ml per tomato plant; once every 3-5 days, 5 times in succession.
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|---|---|---|---|---|
| JP2005232125A (en) * | 2004-02-23 | 2005-09-02 | Mandom Corp | Antiseptic sterilizer, and cosmetic, medicine and food each compounded with the antiseptic sterilizer |
| WO2019159200A1 (en) * | 2018-02-17 | 2019-08-22 | Kanumuru Rahul Raju | Plant micronutrient composition for the management of productivity and disease resistance |
| CN111903456A (en) * | 2020-09-10 | 2020-11-10 | 常州市金坛区植保植检站 | Method for preventing and controlling phytophthora root rot of taros by using potassium silicate solution |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2005232125A (en) * | 2004-02-23 | 2005-09-02 | Mandom Corp | Antiseptic sterilizer, and cosmetic, medicine and food each compounded with the antiseptic sterilizer |
| WO2019159200A1 (en) * | 2018-02-17 | 2019-08-22 | Kanumuru Rahul Raju | Plant micronutrient composition for the management of productivity and disease resistance |
| CN111903456A (en) * | 2020-09-10 | 2020-11-10 | 常州市金坛区植保植检站 | Method for preventing and controlling phytophthora root rot of taros by using potassium silicate solution |
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| Title |
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| Nikky Deepa,et al..Linalool reduces the virulence of Pseudomonas syringae pv. tomato DC 3000 by modulating the PsyI/PsyR quorum-sensing system.Microbial Pathogenesis.2022,第173卷10. * |
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