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Pseudomonas chlororaphis and application thereof in prevention and treatment of tomato stolonidium leaf spot

A technology for leaf spot disease of Pseudomonas aeruginosa and stolonifera, which can be used in applications, bacteria, fungicides, etc., and can solve the problems of drug-resistant pathogenic strains, environmental pollution, chemical residues, etc.

Active Publication Date: 2021-04-30
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the prevention and control measures of tomato diseases rely on chemical control for a long time, but problems such as chemical residues, environmental pollution, and drug-resistant pathogenic strains caused by the large-scale application of synthetic pesticides have become increasingly prominent

Method used

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  • Pseudomonas chlororaphis and application thereof in prevention and treatment of tomato stolonidium leaf spot
  • Pseudomonas chlororaphis and application thereof in prevention and treatment of tomato stolonidium leaf spot
  • Pseudomonas chlororaphis and application thereof in prevention and treatment of tomato stolonidium leaf spot

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1, Isolation, Screening, Identification and Preservation of Bacterial Strain YS05

[0063] 1. Isolation and purification of strain YS05

[0064] The rhizosphere soil samples from the open field crops in Harbin were taken, and 10 g of each soil sample was weighed, added to 90 mL of sterilized water, and placed in a constant temperature shaker at 28 ° C for 30 min. according to 10 -1 -10 -7 For serial dilution, take a dilution gradient of 10 -5 、10 -6 、10 -7 The soil samples were plated and placed in a constant temperature incubator at 28°C for 24 hours. After a single colony grows on the coated plate, use a sterilized toothpick to pick out a single colony of different shapes and line it on the KB solid plate for purification. Place it in a constant temperature incubator at 28°C until a single colony with the same shape grows, and then place it at 4°C Store in the refrigerator for later use. Finally, a total of 236 bacterial strains were isolated from the r...

Embodiment 2

[0097] Embodiment 2, bacterial strain YS05 antibacterial spectrum determination

[0098] Nine common vegetable pathogenic fungi and three pathogenic bacteria were selected to determine the antibacterial spectrum of strain YS05.

[0099] The nine kinds of vegetable pathogenic fungi and three kinds of pathogenic bacteria are shown in Table 4, and the specific sources are as follows:

[0100] Anthracnose (Colletotrichum spp.) has been published in the document "Yu Yang, Shi Yanxia, ​​Fu Junfan, Li Baoju. Mechanism of affinity pathogens to induce disease resistance in cucumber tissue structure. Journal of Plant Protection 2008, (01): 37-42.", the public can Obtained from the Vegetable and Flower Research Institute of the Chinese Academy of Agricultural Sciences to repeat the experiment of this application, and cannot be used for other purposes.

[0101] Botrytis cinerea has been published in the literature "Shi Yanxia, ​​Tang Ming, Jin Zhiwen, Xie Xuewen, Chai Ali, Li Baoju. Eval...

Embodiment 3

[0117] Embodiment 3, bacterial strain YS05 biocontrol factor assay

[0118] 1. Detection of cellulase

[0119] Inoculate 5 μL OD in the center of CMC medium 600 =0.8 strain YS05 bacterial suspension, cultured at 28°C for 24 hours, added 3 mL of Congo red dye for staining for 30 minutes, poured off the dye, then added 5 mL of 1mol / L NaCl solution for decolorization for 15 minutes, and observed the digestion circle. Each treatment was repeated 3 times.

[0120] 2. Detection of protease

[0121] Inoculate 5 μL OD in the center of skim milk medium 600 =0.8 strain YS05 bacteria suspension, observe the digestion circle after culturing at 28°C for 24 hours. Each treatment was repeated 3 times.

[0122] 3. Detection of hydrocyanic acid HCN

[0123] Cut Whatman filter paper into strips of 5mm×25mm; weigh 25mg copper(Ⅱ) ethylacetoacetate and 25mg 4-4'-methylenebis-(N,N-dimethylaniline) respectively, dissolve them in 10mL chloroform, sterilize the Whatman filter The paper strips w...

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Abstract

The invention discloses pseudomonas chlororaphis and application of the pseudomonas chlororaphis in prevention and treatment of tomato stolonidium leaf spot. The pseudomonas chlororaphis is pseudomonas chlororaphis YS05 and is preserved in the general microbiological center of the China Committee for Culture Collection of Microorganisms on July 8, 2020, and the preservation number of the pseudomonas chlororaphis YS05 is CGMCC No.20323. The invention also discloses an application of the pseudomonas chlororaphis YS05 in inhibition of pathogenic bacteria. In bacteriostatic spectrum determination, the strain YS05 shows broad-spectrum resistance and has an antagonistic effect on 9 pathogenic fungi and 3 pathogenic bacteria. In a greenhouse potting control effect determination test, the control effect of the strain YS05 on the tomato Stemphylium leaf spot disease reaches 61.27%. In a test for determining the control effect on the in-vitro leaves of the tomato stolonifer, the control effect of the strain YS05 on the tomato gray leaf spot disease is 71.52%. Therefore, the strain YS05 has a good application prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a strain of Pseudomonas chloropinus and its application in the prevention and treatment of tomato leaf spot disease. Background technique [0002] The pathogens of tomato stalk leaf spot are Stemphylium solani Weber and Stemphylium lycopersici (Enjoji) Yamamoto, which mainly damage tomato leaves and spread to stems and fruits in severe cases. Lesions are small brown dots at the beginning, and gradually expand into a round or nearly round shape. In the later stage of the disease, they are easy to perforate and rupture, causing serious losses. At present, the prevention and control measures of tomato diseases rely on chemical control for a long time, but problems such as chemical residues, environmental pollution, and drug-resistant pathogenic strains caused by the large-scale application of synthetic pesticides have become increasingly prominent. In recent years, biologic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/27A01P1/00A01P3/00A01G7/06C12R1/38
CPCA01N63/27A01G7/06
Inventor 李磊李宝聚黄艺烁谢学文石延霞柴阿丽
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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