A strain of Trichoderma harzianum and its application

Abstract

The invention discloses a strain of Trichoderma harzianum and its application. The Trichoderma strain is Trichoderma harzianum Hb13‑3‑2 CGMCC No.13163. The strain and its metabolites have broad-spectrum inhibitory activity against pathogenic fungi, and confrontation growth experiments show that the Trichoderma strain has the ability to antagonize various pathogenic bacteria such as rubber tree red root bacteria and anthracnose bacteria, and also has the ability to prevent and control other various polluting bacteria and promote The role of rubber seedling growth. Therefore, it can be immediately applied to the cultivation process of rubber tree seedlings, to promote the healthy growth of rubber seedlings, and to shield or inhibit the occurrence of root diseases of rubber trees.

Description

technical field

[0001] The invention relates to a strain of multifunctional Trichoderma harzianum strain and application thereof. Background technique

[0002] Trichoderma Trichoderma Pers. (the sexual stage was once called "Hypocrea Fr.") belongs to Sordariomycetes, Ascomycota, Hypocreaceae, and Hypocreales. This genus of fungi has a wide variety of species and is widely distributed. It is an important class of renewable natural resources with high economic value and application prospects (Doi 1969, 1972; Zhu Zhaoxiang and Zhuang Wenying, 2014). Trichoderma spp. is an important class of multifunctional probiotic fungi that widely exist in nature. They can not only control the infection of pathogenic bacteria and the occurrence of diseases through a variety of mechanisms, but also have the ability to promote crop growth and improve crop resistance to biotic and abiotic stress. They are recognized as environmentally friendly fungi and are used in agriculture, In the fields ...

Examples

Embodiment 1

[0041] Embodiment 1, screening and identification of Trichoderma harzianum (Trichoderma harzianum) Hb13-3-2 bacterial strain

[0042] 1. Screening of Trichoderma strains

[0043] The Hb13-3-2 strain was isolated from the rhizosphere soil of a rubber tree infected with red root disease in the rubber planting area of ​​Danzhou City, Hainan Province. The soil samples of the 5-10cm soil layer of the rubber tree rhizosphere were collected in order of collection places and separate sequentially numbered. Using the dilution plate method, each soil sample was divided by 10 -3 、10 -4 Diluted at different concentrations, inoculated on Bengal red medium (PDA+chloramphenicol 0.015g / L+streptomycin 0.015g / L+Rose Bengal 0.05g / L), cultured at 28°C for 3 days; picked Trichoderma colony and inoculated in On Bengal red medium (without antibiotics), culture at 28°C for 3 days, further isolate and purify single spores, obtain Trichoderma strains, and then number and identify them.

[0044] The...

Embodiment 2

[0062] The biological characteristic of embodiment 2, Hb13-3-2 bacterial strain

[0063] 1. Carbon source and nitrogen source requirement characteristics of Hb13-3-2 strain

[0064] 1. Determination of the optimum growth medium for Trichoderma harzianum Hb13-3-2 CGMCC No.13163

[0065] Comparison of growth of Trichoderma harzianum (Trichoderma harzianum) Hb13-3-2 CGMCC No.13163 on solid medium (pH6) plates as follows:

[0066] Min(Minimal agar medium) medium: KNO 3 10g, KH 2 PO 4 5g, MgSO 4 ·7H 2 O 0.5g, FeCl 3 0.02g, glucose 10g, agar 20g, dilute to 1000mL;

[0067] PDA (potato dextrose) medium: 200 g of peeled potatoes, 20 g of glucose, 14 g of agar, boil the peeled potatoes for 30 minutes and distill the filtrate to 1000 mL with distilled water;

[0068] CMV (corn flour) medium: corn flour 30g, agar 14g, after boiling for 30min, dilute the filtrate to 1000mL with distilled water;

[0069] OA (oat) medium: oatmeal 30g, agar 14g, after boiling for 30min, dilute th...

Embodiment 3

[0094] Embodiment 3, the tolerance of Trichoderma harzianum (Trichoderma harzianum) Hb13-3-2 bacterial strain to special culture conditions

[0095] 1. Salt-alkali tolerance of strain Hb13-3-2

[0096] Preparation contains salt NaCl (mass percentage concentration is respectively 1%, 3%, 5%, 7%, 10% or 15%) or KCl (mass percentage concentration is respectively 1%, 3%, 5%, 7% , 10% or 15%) PDA medium, PDA medium containing alkali portion KOH (mass percentage concentration is respectively 1%, 3%, 5%, 7% or 9%), containing NaOH (mass percentage concentration 1%, 3%, 5%, 7% or 9%) PDA medium or containing Na 2 CO 3 (1%, 3%, 5%, 7% or 9% by mass percentage concentration respectively) PDA medium, used for detecting the tolerance of Hb13-3-2 bacterial strain to different concentrations of saline-alkali.

[0097] A plate of the above-mentioned medium was prepared and inoculated with Hb13-3-2 strain block, and each medium treatment was repeated three times. Incubate in dark at 28°C ...

technical field

[0001] The invention relates to a strain of multifunctional Trichoderma harzianum strain and application thereof. Background technique

[0002] Trichoderma Trichoderma Pers. (the sexual stage was once called "Hypocrea Fr.") belongs to Sordariomycetes, Ascomycota, Hypocreaceae, and Hypocreales. This genus of fungi has a wide variety of species and is widely distributed. It is an important class of renewable natural resources with high economic value and application prospects (Doi 1969, 1972; Zhu Zhaoxiang and Zhuang Wenying, 2014). Trichoderma spp. is an important class of multifunctional probiotic fungi that widely exist in nature. They can not only control the infection of pathogenic bacteria and the occurrence of diseases through a variety of mechanisms, but also have the ability to promote crop growth and improve crop resistance to biotic and abiotic stress. They are recognized as environmentally friendly fungi and are used in agriculture, In the fields ...

Examples

Embodiment 1

[0041] Embodiment 1, screening and identification of Trichoderma harzianum (Trichoderma harzianum) Hb13-3-2 bacterial strain

[0042] 1. Screening of Trichoderma strains

[0043] The Hb13-3-2 strain was isolated from the rhizosphere soil of a rubber tree infected with red root disease in the rubber planting area of ​​Danzhou City, Hainan Province. The soil samples of the 5-10cm soil layer of the rubber tree rhizosphere were collected in order of collection places and separate sequentially numbered. Using the dilution plate method, each soil sample was divided by 10 -3 、10 -4 Diluted at different concentrations, inoculated on Bengal red medium (PDA+chloramphenicol 0.015g / L+streptomycin 0.015g / L+Rose Bengal 0.05g / L), cultured at 28°C for 3 days; picked Trichoderma colony and inoculated in On Bengal red medium (without antibiotics), culture at 28°C for 3 days, further isolate and purify single spores, obtain Trichoderma strains, and then number and identify them.

[0044] The...

Embodiment 2

[0062] The biological characteristic of embodiment 2, Hb13-3-2 bacterial strain

[0063] 1. Carbon source and nitrogen source requirement characteristics of Hb13-3-2 strain

[0064] 1. Determination of the optimum growth medium for Trichoderma harzianum Hb13-3-2 CGMCC No.13163

[0065] Comparison of growth of Trichoderma harzianum (Trichoderma harzianum) Hb13-3-2 CGMCC No.13163 on solid medium (pH6) plates as follows:

[0066] Min(Minimal agar medium) medium: KNO 3 10g, KH 2 PO 4 5g, MgSO 4 ·7H 2 O 0.5g, FeCl 3 0.02g, glucose 10g, agar 20g, dilute to 1000mL;

[0067] PDA (potato dextrose) medium: 200 g of peeled potatoes, 20 g of glucose, 14 g of agar, boil the peeled potatoes for 30 minutes and distill the filtrate to 1000 mL with distilled water;

[0068] CMV (corn flour) medium: corn flour 30g, agar 14g, after boiling for 30min, dilute the filtrate to 1000mL with distilled water;

[0069] OA (oat) medium: oatmeal 30g, agar 14g, after boiling for 30min, dilute th...

Embodiment 3

[0094] Embodiment 3, the tolerance of Trichoderma harzianum (Trichoderma harzianum) Hb13-3-2 bacterial strain to special culture conditions

[0095] 1. Salt-alkali tolerance of strain Hb13-3-2

[0096] Preparation contains salt NaCl (mass percentage concentration is respectively 1%, 3%, 5%, 7%, 10% or 15%) or KCl (mass percentage concentration is respectively 1%, 3%, 5%, 7% , 10% or 15%) PDA medium, PDA medium containing alkali portion KOH (mass percentage concentration is respectively 1%, 3%, 5%, 7% or 9%), containing NaOH (mass percentage concentration 1%, 3%, 5%, 7% or 9%) PDA medium or containing Na 2 CO 3 (1%, 3%, 5%, 7% or 9% by mass percentage concentration respectively) PDA medium, used for detecting the tolerance of Hb13-3-2 bacterial strain to different concentrations of saline-alkali.

[0097] A plate of the above-mentioned medium was prepared and inoculated with Hb13-3-2 strain block, and each medium treatment was repeated three times. Incubate in dark at 28°C ...

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