Isolated Streptomyces DL70 and biocontrol and growth promoting application thereof

A technology of streptomyces and bacteria agent, applied in the promotion of plant growth, streptomyces and its application in the field of plant disease prevention and control, can solve the problems of slow growth rate of streptomyces and low effective number of viable bacteria, and achieve rich strain resources , high number of effective viable bacteria, and the effect of promoting plant growth

Active Publication Date: 2019-11-19
DALIAN UNIV OF TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problems of slow growth rate and low number of effective viable bacteria in the existing Streptomyces, the present invention provides an isolated Streptomyces, and provides the application of the strain in preventing and controlling plant diseases and promoting plant growth.

Method used

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  • Isolated Streptomyces DL70 and biocontrol and growth promoting application thereof
  • Isolated Streptomyces DL70 and biocontrol and growth promoting application thereof
  • Isolated Streptomyces DL70 and biocontrol and growth promoting application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Isolation of Streptomyces sp. DL70

[0026] The Streptomyces sp. DL70 of the present invention is isolated from the deep soil in the rhizosphere of healthy plants on the coast of Dalian, and is obtained after gradient dilution and coating of the soil close to the end of the rhizosphere. The specific operation steps are:

[0027] a. The soil in the rhizosphere of healthy crops in Xinghai Beach, Dalian, was sampled at a depth of 15-20 cm. Weigh 1 g of rhizosphere soil, put it into a Erlenmeyer flask with 99 mL of sterile water added with glass beads, and shake and cultivate at 30° C. and 200 rpm for 60 min.

[0028] b. Perform gradient dilution with a pipette (10 -3 、10 -4 、10 -5 ), take 100 μL of the diluted solution and apply it on the Gaoshi No. 1 medium plate, apply 5 culture dishes for each dilution concentration, and cultivate in a 30° C. incubator for 3-5 days. Observe the growth of actinomycete colonies formed every day, pick the earliest actinomyc...

Embodiment 2

[0031] Example 2: Identification of Streptomyces sp. DL70

[0032] The morphological characteristics of the strain DL70 are as follows: at the optimum culture temperature of 30°C, after 3-5 days of culture on Gao’s No. 1 medium, the colonies formed are round, dense and yellowish, and the hyphae in the base have no septum, no breakage and It is tortoiseshell yellow, the aerial hyphae are branched and pink, the spores are straight or wavy, the spores are oval or round, the surface is smooth, pink and purple when mature. According to the Streptomyces Identification Manual, the strain DL70 has typical morphological characteristics of Streptomyces ( figure 1 ).

[0033] The genomic DNA of the strain DL70 isolated and purified above was extracted, and Sangon Bioengineering (Shanghai) Co., Ltd. carried out 16s rDNA sequencing analysis (see sequence SEQ ID NO.1 for the sequencing results), and BLAST homologous comparison in the NCBI database , selected model strains with high homolo...

Embodiment 3

[0034]Embodiment 3: the preparation of Streptomyces DL70 fermented bacterium liquid and the mensuration of its effective viable count

[0035] 1. Preparation of Streptomyces DL70 fermentation broth

[0036] Using the streptomyces DL70 obtained in Example 2, successively carry out spore suspension preparation, seed cultivation and fermentation cultivation, the specific operation steps are as follows:

[0037] a. Inoculate Streptomyces DL70 on the slant medium under sterile conditions, and after culturing at 30°C for 3 days, use sterile water to collect the spores on the slant medium to prepare a spore concentration of 10 10 Individual / mL spore suspension;

[0038] b. Inoculate the spore suspension in the above step a into the liquid seed medium under aseptic conditions, and culture at 30° C. with shaking at 180 rpm for 24 hours to prepare a liquid seed liquid;

[0039] c. Under sterile conditions, put the liquid seed liquid in the above step b into the liquid fermentation med...

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Abstract

The invention discloses an isolated streptomyces and biocontrol and growth promoting application thereof, and belongs to the field of biotechnology. The Streptomyces sp. DL70 of the invention is deposited in China Center for Type Culture Collection (CCTCC), the deposit number is CCTCC NO.M 2019475, and the deposit date is June20, 2019. The streptomyces is isolated from the deep rhizosphere soil ofhealthy plants along the coastal beach in Dalian, has low nutritional requirements and rapid cell growth, and can achieve an effective number of viable bacteria of not less than 1010 cfu/mL in 48 hours of liquid fermentation. According to the present invention, the Streptomyces DL70 and the fermentation products of the Streptomyces DL70 have highly effective antagonistic effects on a variety of plant pathogenic fungi, significantly promote the rapid growth of plants at the same time, and have obvious advantages of biocontrol and growth promoting application and industrial production potential.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an isolated streptomyces and its application in preventing and treating plant diseases and promoting plant growth. Background technique [0002] Streptomyces is a Gram-positive filamentous actinomycete with high G+C content widely distributed in soil, ocean and extreme environments. The suitable growth temperature is about 30°C and the pH is about 7.0. The metabolic pathway of Streptomyces is complex and can synthesize a variety of secondary metabolites including macrolides, quinones, terpenes, indoles, polyketides, polypeptides, enzymes and lipids. It is used in the production of antibacterial active substances such as antibiotics, auxin, phenylacetic acid, succinic acid and cytokinin and essential regulators of plant growth, so it has important application value in the fields of medical treatment, agriculture, food and environmental protection. Its biological control a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/02A01P3/00A01P21/00C12R1/465
CPCC12N1/20C12R2001/465C12N1/205
Inventor 薛闯吴又多李苗苗
Owner DALIAN UNIV OF TECH
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