A method for indoor identification of sesame seedling wilt

Abstract

The present invention discloses a method for indoor identification of sesame seedling wilt, belonging to the technical field of disease control. The method provided herein uses a Candida lipolytica solution for root irrigation to control sesame plant wilt. This method is an effective, environmentally friendly, and sustainable control method that helps reduce the use of chemical pesticides, improve crop yield and quality, and reduce control costs.

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 world's oldest high-quality oil crops, a source of high-quality edible oil, and a major oil crop in my country. It has a long history of cultivation and is widely distributed, cultivated in all provinces and cities. Sesame fusarium wilt (Sesame Fusarium wilt) is a fungal disease caused by infection with Fusarium oxysporum f.sp. sesami (FOS). It can occur at both the seedling and mature plant stages and is a primary disease in sesame-producing areas in central and northern my country, particularly in Northeast, North China, and Northwest China. The disease is widespread and severe, with an annual incidence rate of 5% to 10%, exceeding 30% in severe cases. This disease often results in immature, shriveled seeds that shatter and fall before harvest, ultimately reducing sesame yields and severely 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 present 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] A second technical solution of the present invention is a drug for preventing and treating plant wilt, 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, comprising applying Candida lipolytica, the drug or the fertilizer to the plant to prevent and treat the wilt disease.

[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 following briefly introduces the drawings required for use in the embodiments. 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 any creative work.

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

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

[0015] Various exemplary embodiments of the present invention are now described in detail. Unless otherwise specified, the methods in the examples are conventional methods. Unless otherwise specified, the reagents used are conventional commercially available reagents or reagents prepared using conventional methods. This detailed description should not be considered a limitation of the present invention, but rather 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 herein are intended only to describe particular embodiments and are not intended to limit the present invention. In addition, for numerical ranges herein, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. The intermediate value within any stated value or stated range, and each smaller range between any other stated value or intermediate value within the stated range, is also encompassed within the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded within the scope.

[0017] Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice 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 any conflict with any incorporated document, the contents 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 described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from the description of the invention. The description and examples are intended to be 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] Unless otherwise specified, the technical solutions described in the present invention are all conventional solutions in the field, and the reagents or raw materials used, unless otherwise specified, are purchased from commercial channels or have been disclosed.

[0021] The embodiments of the present invention provide the use of Candida lipolytica in preventing and treating plant wilt.

[0022] Candida lipolytica may inhibit the growth and reproduction of pathogens by competing for nutrients and space. In soil, complex interactions exist between pathogens and beneficial microorganisms. Candida lipolytica may occupy the ecological niche of pathogens, thereby reducing their attack on plants. During its growth, Candida lipolytica may produce metabolites with antimicrobial activity that can directly inhibit or kill pathogens. These antimicrobial substances may exert their bactericidal effects by destroying pathogen cell walls and membranes or interfering with their metabolic processes. When Candida lipolytica interacts with plant roots, it may trigger plant defense responses and enhance disease resistance. This induced resistance may involve the activation of multiple signaling pathways and defense genes within the plant, thereby enhancing plant resistance to pathogens. Candida lipolytica may also improve the soil microenvironment by decomposing organic matter, promoting nutrient cycling, and regulating soil pH. These changes benefit plant growth and development while hindering the survival and reproduction of pathogens.

[0023] Candida lipolytica, a yeast, is widely found in the natural environment, including soil and plant surfaces. Therefore, the method provided by the present invention of using a Candida lipolytica solution for root irrigation to control sesame plant wilt disease is an effective, environmentally friendly, and sustainable control method that helps reduce the use of chemical pesticides, improve crop yield and quality, and reduce control costs.

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

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

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

[0027] The embodiment of the present invention further 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. with the serial number 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, and Hebei provinces and air-dried indoors.

[0035] (2) Use sterilized scissors to remove an area of about 0.25cm 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. The tissue blocks were transferred to PDA culture plates and cultured at 28°C for 5 days. After observing the white, pink to purple-red aerial hyphae, a few mycelia at the edge of the colony were picked and 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 the Pathogen: Based on the growth of the colony, a small amount of mycelium was transferred to PDB medium and shaken in the dark at 150 rpm and 26°C for 3-5 days. Sample 10-1 was amplified by PCR using ITS primers and sequenced. BLAST analysis was then performed on the NCBI website. The similarity with the Fusarium oxysporum strain was approximately 100%. Therefore, the isolate was confirmed to be Fusarium oxysporum.

[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 cultured at 180 rpm and 30°C for 12 h. The OD value was measured. 600 When the value reaches 1.2 to 1.5, the culture is stopped to obtain the Candida lipolytica solution.

[0038] (5) Sesame plants that were healthy and in basically the same growth state (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-puncture root wound method, and then the roots were irrigated with YPD culture medium, 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-puncture root wound method, and then the roots were irrigated with the Candida lipolytica solution in step (4), 10 mL / plant; the blank group was only irrigated with YPD culture medium, 10 mL / plant.

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

[0041] Level 0 (normal): The plant grows 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 the plant's stem and leaf symptoms.

[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, 3 to 5 sesame fields meeting the following conditions were selected as target fields within the integrated pest control area:

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

[0053] ② Sow earlier than ordinary sesame;

[0054] ③ Apply more fertilizer and the plant grows particularly well;

[0055] ④Continuous sesame cropping 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 and served as the experimental group (three replicates) and the control group (three replicates). Sesame seeds of the same variety were sown in rows with a row spacing of 40 cm and a plant spacing of 10 cm. The experimental group was treated with Candida lipolytica, applied with water at a dosage of 1 kg per plot, twice: once when the sesame had three pairs of true leaves and before flowering.

[0058] Starting from the initial disease outbreak in the field, disease surveys were 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 calculated results showed that the morbidity of the control group was 9.26%, and that of the experimental group was 3.1%. It can be seen that the wilt prevention and control method provided by the present invention can significantly reduce the morbidity of wilt and has good application prospects in agricultural production.

[0061] Obviously, the above embodiments of the present invention are merely examples for the purpose of clearly illustrating the present invention and are not intended to limit the implementation methods of the present invention. A person skilled in the art will be able to make other variations or modifications based on the above description. It is not necessary and impossible to enumerate all implementation methods here. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention shall be included within the scope of protection of the claims of the present invention.

Claims

1. The use of Candida lipolytica in preventing and treating plant wilt disease, characterized in that: The plant wilt disease is sesame wilt caused by Fusarium oxysporum; The Candida lipolytica was obtained from Ningbo Mingzhou Biotechnology Co., Ltd., with the number BMZ124607.

2. A method for preventing and treating plant wilt disease, characterized in that: Apply Candida lipolytica to plants to control wilt; The plant wilt disease is sesame wilt caused by Fusarium oxysporum; The Candida lipolytica was obtained from Ningbo Mingzhou Biotechnology Co., Ltd., with the number BMZ124607.

3. The control method according to claim 2, characterized in that: The application method is: using Candida lipolytica to perform root irrigation treatment on plants.

4. The control method according to claim 2, characterized in that: The administration method is as follows: applying Candida lipolytica with water; The application rate is 1-1.5 kg / mu based on the weight of Candida lipolytica, and it should be applied during the seedling stage and before flowering.

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