阅读说明：本技术 二甲基三硫醚在抑制植物病原菌中的应用 (Application of dimethyl trisulfide in inhibiting plant pathogenic bacteria ) 是由 唐利华 李其利 苏映丹 赵江 郭堂勋 黄穗萍 莫贱友 陈小林 于 2021-09-16 设计创作，主要内容包括：本发明涉及二甲基三硫醚在抑制植物病原菌中的应用。本发明通过室内毒力测定确定了二甲基三硫醚对不同病原菌表现出不同程度的抑制作用,其中对玉米立枯丝核菌和葡萄灰霉病菌抑制效果最好,其次为火龙果溃疡病菌和水稻稻瘟病菌,因此说,二甲基三硫醚可作为所述的植物病原菌的潜在生防制剂。(The invention relates to application of dimethyl trisulfide in inhibiting phytopathogen. The invention determines that the dimethyl trisulfide shows inhibition effects of different degrees on different pathogenic bacteria through indoor toxicity measurement, wherein the inhibition effects on rhizoctonia solani and botrytis cinerea are the best, and the inhibition effects on dragon fruit canker and rice blast germs are the second, so the dimethyl trisulfide can be used as a potential biocontrol preparation for the plant pathogenic bacteria.)

1. Use of dimethyltrisulfide for inhibiting phytopathogenic fungi selected from the group consisting of Asian anthrax (Colletotrichum asiaticum), fruit producing anthrax (Colletotrichum fructicola), Siamese anthrax (Colletotrichum siamensis), Clonorchis cuneata (Colletotrichum clivia), Colletotrichum gloeosporioides (Colletotrichum gloeosporioides), Pyricularia oryzae (Magnaporthe oryzae), Citrus aurantiaca (Penicillium digitatum), Rhizoctonia zeae (Rhizoctonia solani), Phytophthora litchi (Phytophora litchii), Pityrosporum ovale (Neosporotrichum dimyrium), Botrytis cinerea (Botrytis cinerea).

2. The use of dimethyl trisulfide according to claim 1, for inhibiting phytopathogens whose host is mango, satsuma orange, banana, lychee, dragon fruit, grape, rice or corn.

3. The use according to claim 1, wherein the phytopathogen is Pyricularia oryzae (Magnaporthe oryzae), Rhizoctonia solani (Rhizoctonia solani), Rhizoctonia solani (Neospora dimyritum), Botrytis vitis (Botrytis cinerea), and the hosts are rice, corn, dragon fruit, grape, respectively.

4. Use according to claim 1, for fumigating phytopathogens with dimethyl trisulfide.

5. The use of claim 4, wherein the concentration gradient of dimethyl trisulfide used in the fumigation treatment is 0.1-40 μ L/L.

6. The use of claim 5, wherein the concentration gradient of dimethyl trisulfide used in the fumigation treatment is 0.1-20 μ L/L.

7. The use according to claim 1, wherein the phytopathogen is Pyricularia oryzae (Magnaporthe oryzae), Rhizoctonia solani (Rhizoctonia solani), Rhizoctonia solani (Neosporolidium dimyritum), Botrytis vitis (Botrytis cinerea), and the dimethyl trisulfide is used at a concentration gradient of 0.1 μ L/L to 10 μ L/L.

Technical Field

The invention belongs to the field of plant protection, and particularly relates to application of dimethyl trisulfide in inhibiting plant pathogenic bacteria.

Background

Dimethyl trisulfide (DMTS) (also known as dimethyl trisulfide) is a volatile sulfur compound found in leek, onion and other allium species, broccoli, cabbage, cottage cheese, sake and some volatile substances released by bacteria and fungi.

Mango anthracnose is a serious disease of mango, and occurs in mango producing areas around the world, the pathogenic bacteria of the mango are mainly collectible gloeosprioides (Colletotrichum gloospirioides) complex groups, and Asian anthracnose (Colletotrichum asanum), fruit-born anthracnose (Colletotrichum fructicola) and Siamese anthracnose (Colletotrichum size) are 3 dominant species of the mango anthracnose. The disease mainly damages mango leaves, young shoots, inflorescences and fruits and can cause the withered tips, the withered leaves, the fallen flowers and the fallen fruits of mango garden plants and a great amount of rottenness of picked fruits. The black spots appear in the early stage of the disease, then the spots are enlarged to form black round, oval or irregular spots, the spots are usually sunken, red sporophyte is usually generated in the spots under the humid environment, and finally the whole fruit becomes black and decays. The host range of the pathogenic bacteria is wide, besides mango is damaged, various fruits, vegetables, trees, flowers and the like are also damaged, and most tropical and subtropical fruit trees are also important hosts of the pathogenic bacteria.

The rice blast is one of important diseases in global rice production, has great threat to the yield and quality of rice, can cause the complete rice field to be extinct in severe occurrence, the rice blast (Magnaporthe oryzae) is transmitted through conidia, the conidia germinate on the surfaces of rice leaves to generate germ tubes and form attachment cells, then infection plugs are generated to penetrate the cuticle and epidermal cell walls of the leaves, secondary hyphae are generated in host cells in a differentiation manner and infect adjacent cells and tissues, and then the disease is developed.

The orange peel of the granulated sugar is thin, so that the orange peel is easily damaged by machinery in the processes of harvesting, storing and transporting, is easily infected by pathogenic bacteria to cause decay, and has a short storage period after harvesting. Among them, the green mold caused by spores of pathogenic bacteria of Penicillium digitatum (Penicillium digitatum) is one of the main causes of rot after sugar orange harvest.

Fusarium oxysporum (Fusarium oxysporum f.sp. cubense) is a soil-borne pathogenic fungus widely distributed all over the world, has wide pathogenic range and serious pathogenicity, and is listed as one of ten kinds of plant pathogenic fungi in the world. The fusarium oxysporum can infect more than 100 crops with important values to cause blight and root rot, such as tomatoes, bananas, strawberries, watermelons and the like, the fusarium oxysporum infects plants from roots, the plant diseases can be caused in the whole growth period of the plants, the brown necrotic spots are mainly generated on the root cortex of the plants, a main root and a large number of lateral roots are rotted when the plants are serious, and branches are reduced to die of the plants; the vascular tissue of the root of the plant is browned, and the overground part of the plant withers until withering, so that the growth, the yield and the quality of the plant are seriously influenced.

The banana anthracnose causes fruit decay after picking, is one of the most important fungal diseases in banana production, and has 3 dominant pathogenic bacteria of fruit-producing anthracnose bacterium (Colletotrichum fructicola), Clivium basilicum (Colletotrichum cliviticola) and Siamese anthracnose bacterium (Colletotrichum siamense). The banana anthracnose mainly damages mature banana fruits, brown or blackish brown small round spots appear on stems and peel initially, then the spots rapidly expand and are fused with each other, the peel often becomes blackish brown within 2-3 days, the pulp is rotten, and the fruit value is seriously reduced.

The corn sheath blight is a main disease of a main corn production area around the world, a pathogenic bacterium is Rhizoctonia solani (Rhizoctonia solani), the corn sheath blight is mainly harmful to the leaf sheath of corn, the corn sheath blight can be harmful to stems and even ears when the disease condition is serious, the sheath blight becomes a serious disease influencing the corn yield, and great loss is brought to agricultural production.

The litchi frost blight is caused by phytophthora litchi (Peronophythora litchii), can cause browning and rot of litchi fruits, causes yield reduction and yield instability of main diseases, and is commonly generated in litchi cultivation areas in China; the disease is harmful to the young shoots, tender leaves and flower ears of the litchi, can cause economic loss of the litchi industry to be more than 80 percent in the years with suitable weather conditions, and seriously limits the development of the litchi industry.

The dragon fruit canker is one of the most serious diseases in the production management of the current dragon fruits, and the pathogenic bacteria are new phaeocaulus dimyritum (neospora dimyritum). The disease mainly damages the stem of the dragon fruit, leads to stalk rot and fruit cracking in severe cases, even leads to brown rot or black rot of the pulp, and has certain influence on the production of the dragon fruit.

Grape anthracnose is mainly caused by infection of Colletotrichum gloeosporioides, mainly harms grape fruits, and has obvious symptoms near the mature or mature period. After the fruit is damaged, a small brown round spot with the size of a needle head is generated on the surface of the fruit, then the small brown round spot is gradually enlarged and sunken, dark black particles which are concentrically arranged in a ring shape are generated on the surface, namely a conidiophore disk of pathogenic bacteria, pink conidiophore appears at a diseased part when the environmental humidity is high, the diseased spot is expanded to the whole ear when the environment humidity is serious, the diseased ear rate is 50% -70%, and the damage to the grape industry is serious.

Grape gray mold is a common fruit and vegetable disease caused by Botrytis cinerea (Botrytis cinerea) infection, and is also one of the most harmful plant diseases. In addition to tomato, botrytis cinerea can also damage eggplant, pepper, cucumber, grape, strawberry and other important economic crops, and can cause fruit and vegetable botrytis. The disease may occur not only during the host plant growing season but also during storage of the agricultural product.

Disclosure of Invention

In view of the above requirements, the present invention provides the use of dimethyl trisulfide for inhibiting phytopathogens. Indoor toxicity measurement proves that the dimethyl trisulfide has obvious inhibition effect on plant pathogenic bacteria. Dimethyl trisulfide is a potential biocontrol inhibitor, and has high efficiency and low toxicity.

Use of dimethyltrisulfide for inhibiting phytopathogenic fungi selected from the group consisting of Asian anthrax (Colletotrichum asiaticum), fruit producing anthrax (Colletotrichum fructicola), Siamese anthrax (Colletotrichum siamensis), Clonorchis cuneata (Colletotrichum clivia), Colletotrichum gloeosporioides (Colletotrichum gloeosporioides), Pyricularia oryzae (Magnaporthe oryzae), Citrus aurantiaca (Penicillium digitatum), Rhizoctonia zeae (Rhizoctonia solani), Phytophthora litchi (Phytophora litchii), Pityrosporum ovale (Neosporotrichum dimyrium), Botrytis cinerea (Botrytis cinerea).

The host of the plant pathogenic bacteria is mango, sugar orange, banana, litchi, dragon fruit, grape, rice or corn.

The plant pathogenic bacteria are rice blast bacteria (Magnaporthe oryzae), Rhizoctonia solani zea (Rhizoctonia solani), dragon fruit ulcer bacteria (Neoscytalidium dimyridum) and Botrytis cinerea (Botrytis cinerea), and the hosts are rice, corn, dragon fruit and grape respectively.

The application is to utilize dimethyl trisulfide to fumigate plant pathogenic bacteria.

In the fumigation treatment, the concentration gradient of dimethyl trisulfide is 0.1-40 muL/L.

In the fumigation treatment, the concentration gradient of dimethyl trisulfide is 0.1-20 muL/L.

The plant pathogenic bacteria are rice blast fungus (Magnaporthe oryzae), Rhizoctonia solani zea (Rhizoctonia solani), dragon fruit ulcer fungus (Neoscytalidium dimyridum) and Botrytis cinerea (Botrytis cinerea), and the using concentration gradient of the dimethyl trisulfide is 0.1-10 muL/L.

The invention has the beneficial effects that: the dimethyl trisulfide is utilized to carry out fumigation treatment on pathogenic bacteria in a closed environment so as to achieve a bacteriostatic effect, and the inhibition effect of the dimethyl trisulfide on different pathogenic bacteria is determined to different degrees through indoor toxicity measurement, wherein the inhibition effect on rhizoctonia solani and botrytis cinerea is the best, and the inhibition effect on dragon fruit ulcer bacteria and rice blast bacteria are the second, so the dimethyl trisulfide can be used as a potential biocontrol agent for the plant pathogenic bacteria.

Drawings

FIG. 1 shows the colony morphology of (mango) fruit anthrax (Colletotrichum fructicola), Asian anthrax (Colletotrichum asianum), Siamese anthrax (Colletotrichum siamense), and Magnaporthe oryzae (Magnaporthe oryzae) treated with dimethyl trisulfide;

FIG. 2 shows the colony morphology of Citrus viridis (Penicillium digitatum), Fusarium oxysporum (Fusarium oxysporum f.sp.cubense), Colletotrichum fructicola (Colletotrichum fructicola), Colletotrichum roseum (banana) and Colletotrichum clivia after treatment with dimethyl trisulfide;

FIG. 3 shows the colony morphology of Siamese anthrax (banana), Rhizoctonia solani (Rhizoctonia solani), Phytophthora litchi (Phytophora litchii), and Colletotrichum viticola (Colletotrichum gloeosporioides) treated with dimethyl trisulfide;

FIG. 4 shows the colony morphology of Botrytis cinerea (Botrytis cinerea) and Pityrosporum ovale (Neoscytalidium dimidatum) treated with dimethyl trisulfide.

Detailed Description

The present invention will be described in further detail with reference to examples.

Firstly, experimental materials: 1. a compound: dimethyl trisulfide (liquid) purchased from Sigma company, usa.

2. Biological material: the strains are all published except that the strains of citrus green mould (Penicillium digitatum) and Phytophthora litchi (Phytophthora litchii) are obtained by self collection and separation. The strains are preserved in the laboratory of the applicant and can be published and issued to the outside.

II, experimental operation process:

1. the bacterial strains of the plant pathogenic bacteria are respectively cultured on a PDA solid culture medium at 25 ℃ for 5-7 days, and then a hole is punched along the outer edge of the bacterial colony by a puncher with the diameter of 6mm so as to keep the bacterial strains at the same activity.

2. In a large culture dish (volume about 500mL) with diameter of 15cm and height of 3cm, 4 small culture dish bottoms (volume about 35mL) with diameter of 6cm and height of 1.5cm are placed, a tube cover of 12 mL centrifuge tube is placed in the center of the large culture dish, and a circular filter paper sheet with diameter of 15mm multiplied by 15mm is placed in the centrifuge tube cover. Pouring 5mL of PDA culture medium into 4 small culture dishes, inoculating a fungus cake with the diameter of 6mm in the center of the culture dish after the PDA culture medium is solidified, and adding corresponding volumes of dimethyl trisulfide into a centrifuge tube cover in the center of a large culture dish, wherein the corresponding addition amounts of dimethyl trisulfide are 0 muL/L (CK), 1 muL/L (0.5 muL/dish), 5 muL/L (2.5 muL/dish), 10 muL/L (5 muL/dish), 15 muL/L (7.5 muL/dish), 20 muL/L (10 muL/dish) and 40 muL/L (20 muL/dish), and 3 repetitions are carried out for each concentration (namely 3 large dishes) and a group of no dimethyl trisulfide is set as a control. And sealing with a sealing film immediately after the dimethyl trisulfide is added to prevent the dimethyl trisulfide from leaking.

3. After treatment, the mycelia were observed and measured in an incubator at 25 ℃. When the control plate without drug grew to the edge, the colony diameter was measured by the cross method and the inhibition rate was calculated. The inhibition ratio (%) (control hypha diameter-treated hypha diameter)/control hypha diameter × 100 (see table 1). Calculating out virulence regression equation and EC by DPS after obtaining the bacteriostasis rate₅₀And EC₉₅Values (see table 2).

TABLE 1 bacteriostasis rates of dimethyl trisulfide to various pathogenic bacteria at different concentrations%

Remarking: the strains come from different hosts and have slightly different sensitivity to medicaments.

As shown in the table above, when 40 μ L/L of dimethyl trisulfide is used, the inhibition rate of Fusarium oxysporum f.sp.cubense on other plant pathogenic bacteria is above 75% except that the inhibition rate is lower (38.39%); the bacteriostasis rate to a plurality of pathogenic bacteria reaches 100 percent; when 10 mu L/L of dimethyl trisulfide is used, the bacteriostasis rate of the dimethyl trisulfide to rice blast fungus (Magnaporthe oryzae), corn Rhizoctonia solani (Rhizoctonia solani), dragon fruit ulcer fungus (Neoscytalidium dimyritum) and Botrytis cinerea (Botrytis cinerea) can reach 100 percent; particularly, the bacteriostasis rate of 5 mu L/L of dimethyl trisulfide to the Rhizoctonia solani (Rhizoctonia solani) and Botrytis cinerea (Botrytis cinerea) can also reach 100 percent, and particularly the bacteriostasis rate of 1 mu L/L of dimethyl trisulfide to the Rhizoctonia solani (Rhizoctonia solani) also reaches 89 percent.

TABLE 2 regression equation of toxicity of dimethyltrisulfide to various phytopathogens, EC₅₀And EC₉₅Value of

As can be seen from the data calculated in Table 2, the EC50 concentration of dimethyltrisulfide for Rhizoctonia solani (Rhizoctonia solani) was 0.12. mu. L.L^-1EC50 concentration was 0.85. mu. L.L for Botrytis cinerea^-1。