Streptomyces sp. and screening method and application thereof

A screening method and technology of Streptomyces, applied in the field of microbiology, can solve the problems of wide host range, destructive long-term, no chemical control methods, etc., and achieve the effect of significant antagonism

Active Publication Date: 2020-07-03
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the wide geographical distribution of this disease, the wide range of hosts, and its destructive and long-term nature, there is no effective chemical control method at present, and the extensive use of chemical pesticides has brought a series of environmental problems

Method used

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  • Streptomyces sp. and screening method and application thereof
  • Streptomyces sp. and screening method and application thereof
  • Streptomyces sp. and screening method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1. Screening and identification of Streptomyces sp. NEAU-HV9.

[0029] 1. Isolation and cultivation of strains

[0030] The soil samples were collected in Bama Yao Autonomous County, Hechi City, Guangxi Zhuang Autonomous Region, China (24°15′N,107°26′E). 5g of the dried soil samples were dissolved in a 45ml Erlenmeyer bottle of sterile water (with glass beads) , Shake at 180rpm for 30-45 minutes, after standing for 10 minutes, take the upper layer liquid and dilute it by 1000 times, then pipet 200μl of the diluted liquid on HV medium (humic acid 0.1g, MgSO 4 ·7H 2 O 0.05g, KCl 0.17g, Na 2 HPO 4 ·3H 2 O 0.05g, FeSO 4 0.001g, CaCO 3 0.002g, multivitamin 0.1ml, agar 2g, sterile water 100ml) plate medium, placed at 28℃ for 7 days, pick a single colony to inoculate fresh ISP3 plate medium (Oats 2g, K 2 HPO 4 ·3H 2 O 0.05g, MgSO 4 ·7H 2 O 0.02g, KNO 3 0.02g, agar 2g, sterile water 100ml), the purified strain was obtained by repeated transfer, and the purified strain was n...

Embodiment 2

[0046] Example 2. Determination of the antagonistic activity of Streptomyces sp. NEAU-HV9.

[0047] Plant pathogen species and source: Tomato bacterial wilt pathogen Pseudomonas solanacearum is a well-known pathogen in the field that causes tomato bacterial wilt. The strain used in the present invention is provided by the State Key Laboratory of Plant Diseases and Insect Pest Biology, Institute of Plant Protection, Chinese Academy of Agricultural Sciences.

[0048] Preparation of tomato bacterial wilt pathogen suspension: 100 mL of physiological saline (0.9%) was used to wash the bacterial wilt pathogen in the eggplant bottle out of the conical flask for use.

[0049] Antagonistic activity determination: using agar well diffusion method. The specific steps are as follows: inoculate the strains in ISP2 liquid medium, culture in a shake flask to logarithmic phase at 28°C and 250 rpm, and inoculate the seed liquid in the fermentation medium (maltodextrin 4g, Lactose 4g, yeast extract p...

Embodiment 3

[0052] Example 3. Determination of the biological control characteristics of Streptomyces sp.NEAU-HV9 against tomato bacterial wilt at the tomato seedling stage.

[0053] Fill a sterile container with about 15-20ml of bacterial wilt pathogen suspension (OD 600 =0.3), take the sprouted seedlings, set up a total of 3 treatments, the first group, the positive control, the roots of the seedlings are immersed in sterile water for about 30s, then the seedlings are transferred to an empty 1.5 or 2 ml sterile EP tube ; The second group, the negative control, immersed the roots of the seedlings in the bacterial suspension (until the root-stem junction) for about 30 seconds, and then transferred the seedlings to an empty 1.5 or 2 ml sterile EP tube; the third group, In the treatment group, the roots of the seedlings were immersed in the bacterial suspension (until the root-stem junction) for about 30 seconds, and then the roots of the seedlings were respectively immersed in the antagonistic...

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Abstract

The invention discloses a streptomyces sp. and a screening method and application thereof, and belongs to the field of microbiology. In order to improve the resistance of tomatoes to plant diseases, the invention provides a streptomyces sp. strain named NEAU-HV9 with a strain preservation number of CGMCC 19436. The invention also provides application of the strain in inhibiting activity of pathogenic bacteria causing plant diseases and producing actinomycin D, and the streptomyces sp. strain NEAU-HV9 can be used for preparing microbial agents which can be used for inhibiting plant tomato bacterial wilt. The invention provides a high-quality material for the research and development of environment-friendly actinomycetic inoculant.

Description

Technical field [0001] The invention belongs to the field of microbiology, and specifically relates to a Streptomyces and its screening method and application. Background technique [0002] Tomato is one of the most important vegetable crops in the world, with an annual global output of approximately 160 million tons. In my country, long-term continuous cropping is the main cultivation method of tomato, which has caused serious bacterial wilt. It is a soil-borne bacterial disease caused by Ralstonia solanacearum that can cause the whole plant to wilt. Tomato bacterial wilt is not only a serious damage, but also a wide range of areas. It is an important disease for tomato production in tropical, subtropical and temperate regions. Due to the wide geographical distribution of this disease, the wide range of hosts, and its destructive and long-term nature, there is currently no effective chemical control method, and the extensive use of chemical pesticides has brought a series of en...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02A01N63/28A01N43/90A01P1/00C12P21/04C12R1/465
CPCC12N1/02C12N1/20A01N63/00A01N43/90C12P21/02C12R2001/465C12N1/205
Inventor 赵军伟向文胜凌玲王相晶
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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