Streptomyces hydrogenans OsiLf-2 capable of effectively antagonizing magnaporthe oryzae in vitro

A technology of endogenous actinomycetes and rice blast bacteria, applied in the direction of bacteria, biocides, microorganisms, etc., to achieve the effects of inhibiting mycelium growth, lasting drug effect, and strong antifungal activity

Active Publication Date: 2017-02-22
湖南新长山农业发展股份有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the reported disease-resistant endophytes used in the biological control of rice blast are easy-to-cultivate bacteria and fungi, and there are few reports on endophytic actinomycetes that are difficult to isolate
However, as the main source of antibiotics, actinomycetes have greater biocontrol potential

Method used

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  • Streptomyces hydrogenans OsiLf-2 capable of effectively antagonizing magnaporthe oryzae in vitro
  • Streptomyces hydrogenans OsiLf-2 capable of effectively antagonizing magnaporthe oryzae in vitro
  • Streptomyces hydrogenans OsiLf-2 capable of effectively antagonizing magnaporthe oryzae in vitro

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Embodiment 1: Isolation, purification and screening of OsiLf-2 bacterial strain

[0015] A susceptible indica rice variety was taken from Liuyang Daweishan Base of Hunan Yahua Seed Industry Science Research Institute. The leaves with less serious disease were washed, the plant surface was sterilized, dried and placed on mannitol soybean agar medium (MS) at 27 Cultivate at ℃, wait for the growth of endophytic actinomycetes, and then combine the streak separation method to obtain its pure culture on solid potato dextrose agar (PDA) plate. The plate confrontation method was used to inoculate the isolated strains and Magnaporthe grisea on a PDA solid plate at the same time, and put them into a mold incubator at 28°C for upside-down culture. Observed every day, the strain OsiLf-2 with the most obvious antifungal effect was screened out. The inhibition rate against blast fungus mycelium is up to 83%, see attached image 3 .

Embodiment 2

[0016] Example 2: Biological identification of the OsiLf-2 strain

[0017] 1. Strain Morphological Characteristics

[0018] (1) Colony morphology characteristics: Inoculate a single colony of OsiLf-2 into PDA solid medium by streaking, place the plate upside down in a constant temperature incubator, and culture at 28°C. Colorless basal mycelium and aerial mycelium will grow within 3 days , the surface of the colony is smooth and the edges are wrinkled. After 3 days, yellowish-white white spores covered the colony. See attached figure 1 a.

[0019] (2) The morphological characteristics of the bacteria: the bacteria were cultivated by the insert method, and the aerial hyphae and spores were observed under the electron microscope, see the attached figure 1 b.

[0020] 2. OsiLf-2 strain 16S rRNA analysis

[0021] Using the method of Gram-positive Bacteria Genomic DNA Extraction Kit (Shanghai Generay Biotech Co., Ltd), the genomic DNA of the bacterial strain was extracted, a...

Embodiment 3

[0022] Embodiment 3: OsiLf-2 bacterial strain is to the inhibition of different physiological races of Magnaporthe grisea

[0023] Using the plate confrontation method, inoculate the center of a 90mm-diameter petri dish with different physiological races of Magnaporthe grisea. Among them, RB3, 61, 62 and SO7 were from Hunan Yahua Seed Co., Ltd., and Guy11 was from Hunan Academy of Agricultural Sciences. Scrape the activated OsiLf-2 and draw a straight line 30mm away from the pathogenic fungus. Inverted culture at 28°C for more than 7 days, the results are shown in the appendix image 3 , the inhibitory rates of OsiLf-2 against Magnaporthe grisea RB3, 61, 62, Guy11, and SO7 were 74.1%, 74.6%, 82.9%, 83.0%, and 80.6%, respectively.

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Abstract

The invention discloses streptomyces hydrogenans OsiLf-2 capable of effectively antagonizing magnaporthe oryzae in vitro. The strain is collected in CGMCC (China general microbiological culture collection center), with the collection number of CGMCC No. 11673. Streptomyces hydrogenans OsiLf-2 has a good antagonistic effect on magnaporthe oryzae of at least five different physiological races in vitro. Fermentation broth of streptomyces hydrogenans OsiLf-2 also has a relatively high antagonistic effect on magnaporthe oryzae, and the antibacterial activity is very stable. Streptomyces hydrogenans OsiLf-2 and the fermentation broth have great application potential in biological prevention of rice blast, development of novel, safe, nontoxic and environment-friendly pesticides and the like.

Description

technical field [0001] The invention relates to the field of plant endophytic actinomycetes and biological control, in particular to the isolation and resistance-related research of a rice endophytic actinomycete capable of antagonizing rice blast fungus. Background technique [0002] Rice blast is a fungal disease of rice caused by Magnaporthe oryzae. Magnaporthe grisea is one of the most serious diseases of rice due to its strong viability and wide spread. At present, the control of rice blast mainly includes the selection of disease-resistant varieties and chemical control. However, long-term use of chemical pesticides in large quantities pollutes the environment, damages human health, and causes pathogenic bacteria to develop drug resistance. The genetic background of Magnaporthe grisea is complex and easily mutated, which makes it difficult for rice resistant varieties to keep up with the mutation speed of the physiological race of the pathogenic fungus, and the disea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01N63/02A01P3/00C12R1/465
CPCA01N63/00A01N63/10C12N1/205C12R2001/465
Inventor 朱咏华刘选明曾夏冬廖红东刘雨晴徐婷袁珊珊高艳
Owner 湖南新长山农业发展股份有限公司
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