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A Strain of Streptomyces that Produces Penicidin a1 and Inhibits the Growth of Phytopathogenic Xanthomonas

A technology of penidaemycin and phytopathogens, applied in the field of microorganisms, to achieve the effects of simple fermentation conditions, small genome and good results

Active Publication Date: 2022-04-08
周莲
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the fermentative titer of this producing strain to produce penicidin A is only 1.8-1.9 g / L

Method used

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  • A Strain of Streptomyces that Produces Penicidin a1 and Inhibits the Growth of Phytopathogenic Xanthomonas
  • A Strain of Streptomyces that Produces Penicidin a1 and Inhibits the Growth of Phytopathogenic Xanthomonas
  • A Strain of Streptomyces that Produces Penicidin a1 and Inhibits the Growth of Phytopathogenic Xanthomonas

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Obtaining of Streptomyces strain RX1101

[0040] This embodiment provides a method for isolating Streptomyces with antagonism activity against Xanthobacterium oryzae from rice rhizosphere, comprising the steps of:

[0041] Step 1. Isolate rice rhizosphere microorganisms: take a well-grown rice root system in a farmland in Wujing Town, Minhang District, Shanghai suburb, wash it with sterile water to remove surface soil; cut the root system short, and add 50 ml of phosphoric acid to 50 grams of wet weight root system Buffered saline solution (potassium dihydrogen phosphate (KH 2 PO4): 0.24g / L, disodium hydrogen phosphate (Na 2 HPO4): 1.44g / L, Sodium Chloride (NaCl): 8g / l, Potassium Chloride (KCl): 0.2g / L, pH 7.2), milled into a slurry; after filtering with gauze, take 2ml of the filtrate Perform 2-fold, 10-fold and 100-fold dilutions, and each take 100 microliters of the dilution and spread it on the "Gao Shi No. 1" agar plate (soluble starch 20g / L, KNO 3 1g...

Embodiment 2

[0044] Example 2 Gene sequence analysis of Streptomyces strain RX1101

[0045] 1. Genomic DNA extraction and purification of Streptomyces strain RX1101: RX1101 strain was inoculated in NA liquid medium and cultured at a constant temperature of 28°C for 36 hours, and 5 ml of bacterial liquid was collected by centrifugation. Whole-genome DNA was extracted using Bacterial DNA Kit (OMEGA bio-tek), dissolved in 100 µl sterile water, with a final concentration of 120 ng / µl.

[0046] 2. Analysis of the 16s rRNA gene sequence of RX1101 strain: using primers 16S-FOR: ACCATTCCGCAGCTGC (SEQ ID NO.2) and 16S-REV: GCTATGACCATGATTACGG (SEQ ID NO.3), using the RX1101 genomic DNA extracted in step 1 as a template, Amplification was performed using the PCR program described below.

[0047] The PCR program is as follows:

[0048] Pre-denaturation at 1.94°C for 5 minutes,

[0049] Denaturation at 2.94℃ for 30s,

[0050] Annealing at 3.55℃ for 30s,

[0051] Extend at 4.72°C for 40s, repeat t...

Embodiment 3

[0060] Liquid culture and metabolite extraction of embodiment 3 Streptomyces strain RX1101

[0061] 1. Pick a single colony of RX1101 with a toothpick, inoculate it in 50 ml of NA liquid medium, and culture it in a constant temperature shaker at 28°C for 48 h, with the shaker speed at 200 rpm. Add an equal volume of ethyl acetate, shake and extract for 5 minutes, let it stand for 10 minutes, transfer about 40 ml of ethyl acetate from the upper layer to another centrifuge tube; evaporate the ethyl acetate to dryness with a rotary distillation apparatus, add 1 ml of methanol to dissolve Extracts were used for further analysis of antibacterial activity.

[0062] 2. Identification of the activity of RX1101 metabolites in inhibiting the growth of Xanthomonas oryzae PXO99A: After diluting the crude methanol extract obtained above 10 times, 100 times, 1000 times and 10000 times, take 10 microliters and slowly drop it on a sterile circle Shaped filter paper, after drying in the ultra...

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Abstract

The invention discloses a Streptomyces strain with high-yield pieridin and high-efficiency inhibition of the growth of phytopathogenic Xanthomonas. The name of the strain is Streptomyces RX1101. NO: M 2020069. The genotype and phenotype of the Streptomyces strain were first discovered. The strain has a small genome, simple fermentation conditions, and the fermentation time of the target metabolite is only 3 days, which is far shorter than that of traditional Streptomyces. The Streptomyces strain highly produces a fat-soluble metabolite, Penicidin A1, which can effectively inhibit the growth of various phytopathogenic Xanthomonas, and the fermentation titer of its production of Penicidin A1 reaches 2.65 g / L, which is far It is much higher than the fermentation titer of other bacterial strains piericidin reported so far, which is more conducive to industrialization.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a streptomyces strain, in particular to a streptomyces strain which produces penicidin A1 and inhibits the growth of phytopathogenic xanthomonas. Background technique [0002] Streptomyces (Streptomyces) is a very large genus in the Bacteria Kingdom, Firmicutes, Actinomycetes, Actinomycetes, Actinomycetes, Streptomycetes, and Streptomycetaceae, and is a typical representative of Actinomycetes . Widely distributed in soils with low water content, good ventilation, rich in organic matter and slightly alkaline. Gram-positive, the mycelium is developed, without separation, and exists in two states: the aerial hyphae that stretch in the space and the mycelium that is thinner, branched, and does not break. Aerobic bacteria, chemoorganotrophic type, obtain energy through aerobic respiration. It has low nutritional requirements and can utilize a variety of carbon so...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P17/12A01N63/28A01P3/00C12R1/465
CPCC12N1/20C12P17/12A01N63/28C12R2001/465C12N1/205
Inventor 何亚文
Owner 周莲
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