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Trichoderma protoplast fusion strain and application thereof

A protoplast fusion, Trichoderma technology, applied in the application, biocide, microorganism and other directions, can solve the problems of restricting popularization and application, the field effect of Trichoderma preparations is not ideal, etc., and achieves high control effect, excellent biocontrol properties, and large yield. Effect

Active Publication Date: 2017-02-22
山东济清科技服务有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the unsatisfactory field effect of Trichoderma preparations from natural sources, its further application is restricted.

Method used

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  • Trichoderma protoplast fusion strain and application thereof
  • Trichoderma protoplast fusion strain and application thereof
  • Trichoderma protoplast fusion strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1 Trichoderma fusion son Tpf-2

[0021] The preservation number of the Trichoderma fusion son tpf-2 of the present invention is CGMCC No.12477, the preservation date is May 30, 2016, the classification name is Trichoderma sp., and the reference microbial strain is Tpf-2. The preservation unit is the General Microbiology Center of the China Committee for the Collection of Microbial Cultures, and the address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The invention is formed by fusion of Trichoderma harzianum T. harzianum TW21990 and Trichoderma paracyanodichotomus TW21990-1 through protoplast fusion.

Embodiment 2

[0022] Embodiment 2 protoplast fusion

[0023] The parental strains used in the experiment were T.harzianum TW21990 and T.cyanodichotomus TW21990-1. After the young mycelia of the two kinds of Trichoderma were treated with a pretreatment agent, the mycelium of TW21990 was lysed with Lysing Enzyme, and the mycelium of TW21990-1 was lysed with Driselase. Filter with a layer of lens paper to remove residual mycelia, collect protoplasts by centrifugation and wash with STC, and adjust the protoplast volume by 10 6 individual / mL.

[0024] The biological strain preservation number of TW21990 is: CGMCC No.12864, and the preservation date is: September 1, 2016. The preservation unit is the General Microbiology Center of the China Microbiological Culture Collection Management Committee, and the address is No. 1 Beichen West Road, Chaoyang District, Beijing Hospital No. 3, the taxonomic name is Trichoderma harzianum.

[0025] The biological strain preservation number of TW21990-1 is: ...

Embodiment 3

[0035] Embodiment 3 Fusion screening

[0036] Determination of the biocontrol properties of the fusion, mainly including the inhibition of Pythium melon and Pythium ultima on the PDA plate; the width of the hydrolysis band produced on the enzyme activity plate, including β-1,3-glucanase, Cellulase and protease; yield of chlamydospores in liquid fermentation. The gray correlation degree analysis method was used to comprehensively evaluate the fusions obtained from the preliminary screening, and the excellent fusions were determined according to the correlation degree.

[0037] Inhibition rate%=100%×(control bacteria growth radius-treatment bacteria growth radius) / control bacteria growth radius

[0038] 50×Trace elements (L): FeS0 4 ·7H 2 0 0.25g, ZnSO 4 2H 2 O 0.07g, CoCl 2 ·6H 2 O 0.1g, MnSO 4 ·H 2 O 0.085g.

[0039] β-1,3-glucanase medium (L): glucose 20g, yeast extract 0.67g, K 2 HPO 4 15g, (NH 4 ) 2 S0 4 5g, CaCl 2 0.6g, MgCl 2 0.6g, 50×trace elements 20...

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Abstract

The invention relates to a trichoderma protoplast fusion strain and application thereof. The trichoderma protoplast fusion strain is a trichoderma fusant Tpf-2, wherein the preservation number is CGMCC No.12477, the preservation date is May 30, 2016, the preservation unit is China General Microbiological Culture Collection Center, and the address is 3#, yard No.1, Bechen west road, Chaoyang District, Beijing. The trichoderma protoplast fusion strain can effectively prevent and treat diseases caused by pythium aphanidermatum and pythium wltmum, the prevention and treatment effect on tomato damping-off caused by the pythium aphanidermatum and the pythium wltmum reaches 86.3 percent and 85.7 percent separately, the yield-increasing effect reaches 77.3 percent and 79.9 percent separately (p is less than 0.01), the survival rate of the trichoderma protoplast fusion strains which are colonized in tomato rhizosphere soil for five months reaches 75 percent, and the effective survival rate of chlamydospore preparations within 1-year guarantee period reaches 90 percent.

Description

technical field [0001] The present invention relates to a Trichoderma protoplast fusion strain and its application, in particular to a Trichoderma fusion obtained by fusion of heat-ultraviolet inactivated protoplasts and the application of the fusion in the control of tomato Pythium damping-off, Belongs to the field of biotechnology. Background technique [0002] Tomato is the main vegetable crop in my country. Various diseases and insect pests often occur during the planting process. Among them, tomato damping-off is an important disease of tomato, which mostly occurs in the seedling stage and leads to the collapse of seedlings. Pythium melon and fruit P.aphanidermatum and Pythium ultimum P.ultimum, this type of pathogen is a lower fungus of the genus Pythium in the subphylum Flagellate of Oomycota, with a fast growth rate, often saprophytic or parasitic in soil and water Survive for a long time and bring serious losses to agricultural production. [0003] The bacterial age...

Claims

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Application Information

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IPC IPC(8): C12N1/14A01N63/04A01P3/00C12R1/885
CPCA01N63/30C12N1/145C12R2001/885
Inventor 陈凯李纪顺魏艳丽王贻莲扈进冬李玲赵忠娟杨合同
Owner 山东济清科技服务有限公司
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