Trichoderma strain for antagonizing soil-borne disease

Abstract

The invention relates to a Trichoderma strain for antagonizing soil-borne disease, belonging to the field of biological control. The Trichoderma strain is Trichoderma asperellum ZJSX5003 CGMCC No.6480. The Trichoderma strain provided by the invention has the characteristics of high growth speed and high spore production amount; the in-vitro standoff pathogen inhibition rates are respectively up to 79.79%, 73.68% and 72.25%, the pathogen spore survival rates are respectively 0.00%, the in-vivo preventive effects for cucumber fusarium wilt and maize stem rot are respectively 85.08% and 66.67%, the enzyme activity of chitinase is 4.0783U, the enzyme activity of beta-1,3-glucanase is 0.8050U, the enzyme activity of extracellular proteinase is 4.8503U, and the content of silicone, terpene and carboxylic acid having antagonistic actions in antibiotic secondary metabolites is 44.44%. The Trichoderma strain provided by the invention has the advantages of high antagonism and high specificity. The biological pesticide produced from the strain can effectively control the soil-borne disease, thereby ensuring the stable increase of crop and vegetable yields.

Description

technical field

[0001] The invention belongs to the field of biological control, and relates to a Trichoderma strain, in particular to a Trichoderma strain antagonizing soil-borne diseases. Background technique

[0002] Due to the long-term planting of a certain crop year after year, serious secondary salinization of the soil and aggravation of soil-borne diseases have resulted in reduced crop quality and reduced yield, which has become an important obstacle restricting the sustainable development of facility agriculture. Plant soil-borne diseases are an important class of plant diseases. There are many kinds of pathogens (fungi, bacteria, nematodes, viruses) that cause soil-borne diseases. They usually infect the roots of plants, causing diseases of plant roots and even the whole plant, causing major Economic losses. Among them, soil-borne pathogenic fungi have become one of the main restrictive factors for the stable and high yield of crops and vegetables.

[0003] Takin...

Examples

Embodiment 1

[0019] Example 1 , Antibacterial test in vitro

[0020] Flat plate confrontation culture method: Use a sterilized Φ6mm puncher to punch out Trichoderma and pathogenic bacteria (Fusarium moniliforme, Fusarium graminearum, Rhizoctonia solani) for about 4 days. Place on one side of a petri dish with a diameter of 90 mm, and inoculate Trichoderma bacteria on the other side, with a distance of 4 cm between the two bacteria plates, and use a single pathogenic fungus as a control, 3 replicates, and culture at a constant temperature of 28 °C. After 5 days of culture, the diameters of pathogenic fungal colonies in the treatment group and the control group were measured by the cross method, and the inhibition rates after 5 days of culture were calculated.

[0021] Colony growth diameter (mm) = average value of colony diameter - 6.0mm;

[0022] Mycelial growth inhibition rate (%) = (control colony growth diameter - treatment colony growth diameter) / control colony growth diameter × 1...

Embodiment 2

[0026] Example 2 , Determination of the survival rate of pathogenic spores

[0027] Through the full contact of pathogenic bacteria (Fusarium oxysporum, Fusarium graminearum, Rhizoctonia solani) with the spore suspension of biocontrol bacteria, four pieces of 5mm-diameter bacteria cakes were taken from the edge of the activated pathogenic bacteria for 5 days, and the Dip into Trichoderma spore suspension (10 6 cells / mL) for 30s, and then cultured overnight in a petri dish covered with 3 layers of wet filter paper. The sterile water was used to replace the Trichoderma spore suspension as a control, and the next day, the pathogenic bacteria cake was transferred to a culture medium containing 1 mg / L benzene In the prodazim selective medium, cultivate in a constant temperature incubator at 28°C, and repeat 3 times. The test results were recorded on the 5th day, and finally expressed in the growth diameter and survival rate of pathogenic bacteria, and the average value was taken...

Embodiment 3

[0031] Example 3 , Greenhouse live control test

[0032] Activate pathogenic bacteria and Trichoderma in PDA medium and culture them at 28°C for 7 days, rinse the spores of pathogenic bacteria on the medium with sterile water, filter the bacterial solution with sterilized filter paper to remove hyphae, and add appropriate amount of distilled water to the filtrate for pathogenic bacteria Dilute to a spore concentration of 1×10 6 individual / mL suspension, Trichoderma dilute the filtrate with appropriate distilled water to a spore concentration of 1×10 7 Individual / mL suspension, set aside.

[0033] (1) Cucumber Fusarium wilt control experiment

[0034] The surface-sterilized cucumber seeds were germinated in a petri dish with double-layer wet filter paper, 6 seeds per dish, and repeated 3 times. Moisturize and cultivate with sterile water for several days. After the seeds germinate and remove the seed coat, plant cucumber seedlings in pots in the greenhouse. When the cucumber...

technical field

[0001] The invention belongs to the field of biological control, and relates to a Trichoderma strain, in particular to a Trichoderma strain antagonizing soil-borne diseases. Background technique

[0002] Due to the long-term planting of a certain crop year after year, serious secondary salinization of the soil and aggravation of soil-borne diseases have resulted in reduced crop quality and reduced yield, which has become an important obstacle restricting the sustainable development of facility agriculture. Plant soil-borne diseases are an important class of plant diseases. There are many kinds of pathogens (fungi, bacteria, nematodes, viruses) that cause soil-borne diseases. They usually infect the roots of plants, causing diseases of plant roots and even the whole plant, causing major Economic losses. Among them, soil-borne pathogenic fungi have become one of the main restrictive factors for the stable and high yield of crops and vegetables.

[0003] Takin...

Examples

Embodiment 1

[0019] Example 1 , Antibacterial test in vitro

[0020] Flat plate confrontation culture method: Use a sterilized Φ6mm puncher to punch out Trichoderma and pathogenic bacteria (Fusarium moniliforme, Fusarium graminearum, Rhizoctonia solani) for about 4 days. Place on one side of a petri dish with a diameter of 90 mm, and inoculate Trichoderma bacteria on the other side, with a distance of 4 cm between the two bacteria plates, and use a single pathogenic fungus as a control, 3 replicates, and culture at a constant temperature of 28 °C. After 5 days of culture, the diameters of pathogenic fungal colonies in the treatment group and the control group were measured by the cross method, and the inhibition rates after 5 days of culture were calculated.

[0021] Colony growth diameter (mm) = average value of colony diameter - 6.0mm;

[0022] Mycelial growth inhibition rate (%) = (control colony growth diameter - treatment colony growth diameter) / control colony growth diameter × 1...

Embodiment 2

[0026] Example 2 , Determination of the survival rate of pathogenic spores

[0027] Through the full contact of pathogenic bacteria (Fusarium oxysporum, Fusarium graminearum, Rhizoctonia solani) with the spore suspension of biocontrol bacteria, four pieces of 5mm-diameter bacteria cakes were taken from the edge of the activated pathogenic bacteria for 5 days, and the Dip into Trichoderma spore suspension (10 6 cells / mL) for 30s, and then cultured overnight in a petri dish covered with 3 layers of wet filter paper. The sterile water was used to replace the Trichoderma spore suspension as a control, and the next day, the pathogenic bacteria cake was transferred to a culture medium containing 1 mg / L benzene In the prodazim selective medium, cultivate in a constant temperature incubator at 28°C, and repeat 3 times. The test results were recorded on the 5th day, and finally expressed in the growth diameter and survival rate of pathogenic bacteria, and the average value was taken...

Embodiment 3

[0031] Example 3 , Greenhouse live control test

[0032] Activate pathogenic bacteria and Trichoderma in PDA medium and culture them at 28°C for 7 days, rinse the spores of pathogenic bacteria on the medium with sterile water, filter the bacterial solution with sterilized filter paper to remove hyphae, and add appropriate amount of distilled water to the filtrate for pathogenic bacteria Dilute to a spore concentration of 1×10 6 individual / mL suspension, Trichoderma dilute the filtrate with appropriate distilled water to a spore concentration of 1×10 7 Individual / mL suspension, set aside.

[0033] (1) Cucumber Fusarium wilt control experiment

[0034] The surface-sterilized cucumber seeds were germinated in a petri dish with double-layer wet filter paper, 6 seeds per dish, and repeated 3 times. Moisturize and cultivate with sterile water for several days. After the seeds germinate and remove the seed coat, plant cucumber seedlings in pots in the greenhouse. When the cucumber...

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