Indoor identification method for fusarium wilt in sesame seedling stage

Abstract

The invention discloses a method for indoor identification of fusarium wilt in a sesame seedling stage, and belongs to the technical field of disease control. The method for preventing and treating the sesame plant fusarium wilt by using the candida lipolytica liquid for root irrigation is an effective, environment-friendly and sustainable prevention and treatment method, and is beneficial to reducing the use of chemical pesticides, improving the yield and quality of crops and reducing the prevention and treatment cost at the same time.

Description

Technical Field

[0001] The invention relates to the technical field of disease prevention and control, in particular to a method for indoor identification of sesame seedling wilt. Background Art

[0002] Sesame (Sesamumindicum L.) is one of the oldest high-quality oil crops in the world, a high-quality edible oil source, and one of the main oil crops in my country. It has a long history of cultivation and is widely distributed in all provinces and cities. Sesame Fusarium wilt is a fungal disease caused by infection with Fusarium Oxysporum f.sp. sesami (FOS). It can occur in both the seedling and adult stages and is the primary disease in sesame producing areas in central and northern my country, especially in sesame producing areas such as Northeast China, North China, and Northwest China. The disease is widespread and serious, with an annual incidence rate of 5% to 10%, and can reach more than 30% when the disease is severe. It often causes immature sesame seeds, thin and shriveled grains, and explodes before harvest, ultimately causing sesame yield reduction, seriously threatening and hindering sesame production and development.

[0003] Therefore, improving the wilt resistance of sesame plants is crucial in agricultural production. Summary of the invention

[0004] The purpose of the invention is to provide a method for indoor identification of sesame seedling wilt disease to solve the problems existing in the above-mentioned prior art.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] One of the technical solutions of the present invention is the use of Candida lipolytica in preventing and treating plant wilt disease.

[0007] The second technical solution of the present invention is a drug for preventing and treating plant wilt disease, comprising Candida lipolytica.

[0008] A third technical solution of the present invention is a fertilizer for preventing and treating plant wilt, comprising Candida lipolytica.

[0009] A fourth technical solution of the present invention is a method for preventing and treating plant wilt disease, wherein the wilt disease is prevented and treated by applying Candida lipolytica, the drug or the fertilizer to the plant.

[0010] Based on the above technical solution, the present invention has the following technical effects:

[0011] The method provided by the present invention for preventing and controlling sesame plant wilt disease by root irrigation using a Candida lipolytica liquid is an effective, environmentally friendly and sustainable prevention and control method, which helps to reduce the use of chemical pesticides, improve crop yield and quality, and reduce prevention and control costs. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0013] Figure 1 This is the plant growth situation of the field experimental group.

[0014] Figure 2 This is the growth condition of diseased plants in the field control group. DETAILED DESCRIPTION

[0015] Now, various exemplary embodiments of the present invention are described in detail. The methods in the examples are conventional methods unless otherwise specified, and the reagents used are conventional commercially available reagents or reagents prepared by conventional methods unless otherwise specified. This detailed description should not be considered as a limitation of the present invention, but should be understood as a more detailed description of certain aspects, features and embodiments of the present invention.

[0016] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0017] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0018] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to the skilled artisan. The present invention description and examples are exemplary only.

[0019] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0020] The technical solutions described in the present invention, unless otherwise specified, are all conventional solutions in the art, and the reagents or raw materials used, unless otherwise specified, are purchased from commercial channels or have been disclosed.

[0021] The embodiment of the present invention provides the use of Candida lipolytica in preventing and treating plant wilt disease.

[0022] Candida lipolytica may inhibit the growth and reproduction of pathogens by competing for nutrients and space resources. In the soil, there are complex interactions between pathogens and beneficial microorganisms. Candida lipolytica may reduce the damage of pathogens to plants by occupying the ecological niche of pathogens. Candida lipolytica may produce metabolites with antimicrobial activity during its growth, which can directly inhibit or kill pathogens. These antimicrobial substances may exert their bactericidal effects by destroying the cell wall and cell membrane of pathogens or interfering with their metabolic processes. When Candida lipolytica interacts with plant roots, it may trigger the plant's defense response and improve the plant's disease resistance. This induced resistance may involve the activation of multiple signal transduction pathways and defense genes in plants, thereby enhancing the plant's resistance to pathogens. Candida lipolytica may also improve the soil microenvironment in the soil by decomposing organic matter, promoting nutrient cycling, and regulating soil pH. These changes are beneficial to the growth and development of plants, but are also not conducive to the survival and reproduction of pathogens.

[0023] As a yeast, Candida lipolytica is widely present in the natural environment, including soil, plant surface, etc. Therefore, the method of using Candida lipolytica liquid to irrigate the roots to prevent and control wilt of sesame plants provided by the present invention is an effective, environmentally friendly and sustainable prevention and control method, which helps to reduce the use of chemical pesticides, improve crop yield and quality, and reduce prevention and control costs.

[0024] In some specific embodiments, the plant comprises sesame.

[0025] The embodiment of the present invention also provides a drug for preventing and treating plant wilt disease, comprising Candida lipolytica.

[0026] The embodiment of the present invention also provides a fertilizer for preventing and treating plant wilt, comprising Candida lipolytica.

[0027] The embodiment of the present invention also provides a method for preventing and treating plant wilt disease, wherein Candida lipolytica, the drug or the fertilizer is applied to the plant to prevent and treat the wilt disease.

[0028] In some specific embodiments, the application method is: using Candida lipolytica, the drug or the fertilizer to perform root irrigation treatment on plants.

[0029] In some specific embodiments, the administration is carried out by: applying the Candida lipolytica, the drug or the fertilizer with water;

[0030] The application rate is 1-1.5 kg / mu based on the weight of Candida lipolytica, and it is applied during the seedling stage and before flowering.

[0031] The Candida lipolytica used in the present invention was purchased from Ningbo Mingzhou Biotechnology Co., Ltd. and is numbered BMZ124607.

[0032] Example 1

[0033] 1. Isolation and identification of pathogenic bacteria of Fusarium wilt

[0034] (1) Sesame plants infected with Fusarium wilt were collected from Henan, Jiangxi, Anhui, Hubei, Hebei and other places and dried naturally indoors;

[0035] (2) Use sterile scissors to remove an area of ​​about 0.25 cm 2 Diseased tissues of different sizes were immersed in 75% alcohol solution and 0.1% mercuric chloride solution for disinfection for 20 seconds and 3 minutes respectively, then rinsed with sterile water and transferred to PDA medium plates. After culturing at 28°C for 5 days, white, pink to purple-red aerial hyphae colonies were observed. A small amount of mycelium at the edge of the colonies was transferred to a new PDA plate for further separation and purification, and cultured at 28°C for 3-4 days (Yang Huihui et al. Identification and biological characteristics of a sesame wilt pathogen. Chinese Journal of Oil Crops, 2014, 36(3): 385-392.).

[0036] (3) Molecular biological identification of pathogens: According to the growth status of the colony, a small amount of mycelium was selected and transferred to PDB medium, and the culture was shaken in the dark at 150 rpm and 26°C for 3-5 days. Sample 10-1 was PCR amplified and sequenced using ITS primers, and then logged on to the NCBI website for BLAST comparison analysis. After comparison, the similarity with Fusarium oxysporum strain was basically 100%. It can be determined that the isolated Fusarium oxysporum is.

[0037] (4) Candida lipolytica was inoculated into YPD culture medium (yeast extract powder 10 g / L, peptone 20 g / L, glucose 20 g / L, pH 5.0-6.0) for activation culture. The culture was carried out at 180 rpm and 30°C for 12 h. The OD 600 When the value reaches 1.2 to 1.5, the culture is stopped to obtain the Candida lipolytica solution.

[0038] (5) Healthy sesame plants with basically the same growth status (three pairs of true leaves) were randomly divided into three groups. The culture medium was sterile vermiculite and nutrient soil, namely the model group, the treatment group and the blank group, with 50 plants in each group.

[0039] The model group was inoculated with a 10-1 spore suspension of the Fusarium oxysporum sample identified in step (3) by the needle-piercing root method, and then the roots were irrigated with YPD culture solution, 10 mL / plant; the treatment group was inoculated with a 10-1 spore suspension of the Fusarium oxysporum sample identified in step (3) by the needle-piercing root method, and then the roots were irrigated with the Candida lipolytica solution in step (4), 10 mL / plant; the blank group was only treated with root irrigation with YPD culture solution, 10 mL / plant.

[0040] According to the symptoms of wilt in the seedling stage, the occurrence level of sesame seedling wilt is divided into 0 to 4 levels:

[0041] Level 0 (normal): The plant is growing well and has no symptoms, the aboveground stems and leaves are normal, and the vascular bundles of the rhizomes are normal; Level 1: A few leaves of the plant wilt, the aboveground stems and leaves are normal, and the vascular bundles of the rhizomes are normal; Level 2: Some leaves of the plant wilt, the stems are slightly constricted, and the vascular bundles of the rhizomes are normal; Level 3: Most leaves of the plant wilt, the stems are constricted, and reddish-brown spots appear on the epidermis of the rhizomes; Level 4 (death): All leaves turn yellow and dry up, the whole plant wilts, and it falls over and dies.

[0042] During the investigation, the disease index of each treatment can be calculated based on the disease occurrence level of stem and leaf symptoms of the plants.

[0043] The calculation formula of disease index (DI) is as follows:

[0044]

[0045] The calculation results of the disease index are shown in Table 1.

[0046] Table 1 Calculation results of disease index

[0047]

[0048]

[0049] As shown in Table 1, root irrigation with Candida lipolytica solution can effectively prevent the occurrence of sesame plant wilt disease. Compared with chemical pesticides, this biological control method has less impact on the environment and is a more environmentally friendly control method.

[0050] Example 2

[0051] According to the provisions of Appendix A of NY / T 2043-2011, select 3 to 5 sesame fields that meet the following conditions as target fields in the integrated control area:

[0052] ① Planting early-maturing and disease-susceptible varieties;

[0053] ② Sowing earlier than ordinary sesame;

[0054] ③ Apply more fertilizer, and the growth is particularly good;

[0055] ④Continuous sesame planting on dry land;

[0056] ⑤Sesame fields with low terrain and poor field drainage.

[0057] Divide the selected plot into six plots of equal size, each with an area of ​​600m 2 The plots were spaced 1 meter apart, serving as the experimental group (3 replicates) and the control group (3 replicates). The same variety of sesame was sown in row sowing, with a row spacing of 40 cm and a plant spacing of 10 cm. The experimental group was treated with Candida lipolytica, which was applied with water at a dosage of 1 kg / time / plot, and applied twice, when sesame grew 3 pairs of true leaves and before flowering.

[0058] Starting from the initial disease period in the field, the disease survey was conducted every 7 days, the number of plants with disease during the survey period was counted, and the incidence rate was calculated:

[0059] Incidence rate = number of diseased plants / total number of plants × 100%.

[0060] The final calculation result is that the morbidity of the control group is 9.26%, and the morbidity of the experimental group is 3.1%. It can be seen that the wilt disease prevention and control method provided by the present invention can significantly reduce the morbidity of wilt disease and has good application prospects in agricultural production.

[0061] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation methods of the present invention. For ordinary technical users in the relevant field, other different forms of changes or modifications can be made on the basis of the above description. It is not necessary and impossible to list all the implementation methods here. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. Application of Candida lipolytica in preventing and controlling plant wilt diseases.

2. The use according to claim 1, characterized in that: The plants include sesame.

3. A drug for preventing and treating plant wilt, characterized in that: Includes Candida lipolytica.

4. A fertilizer for preventing and treating plant wilt, characterized in that: Includes Candida lipolytica.

5. A method for preventing and treating plant wilt disease, characterized in that: Candida lipolytica, the drug according to claim 3 or the fertilizer according to claim 4 are applied to plants to prevent and control wilt disease.

6. The control method according to claim 5, characterized in that: The application method is: using Candida lipolytica, the medicine according to claim 3 or the fertilizer according to claim 4 to perform root irrigation treatment on plants.

7. The control method according to claim 5, characterized in that: The method of application is: applying the Candida lipolytica, the drug according to claim 3 or the fertilizer according to claim 4 with water; The application rate is 1-1.5 kg / mu based on the weight of Candida lipolytica, and it is applied during the seedling stage and before flowering.

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