Porphyra filament albinism pathogenic bacteria

A technology for albinism and filaments, applied in the direction of bacteria, climate change adaptation, biochemical equipment and methods, etc., can solve problems such as unresolved causes of disease

Active Publication Date: 2021-07-02
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] So far, Porphyra filamentous albinism has been reported for the first time, and its cause has not been resolved

Method used

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  • Porphyra filament albinism pathogenic bacteria
  • Porphyra filament albinism pathogenic bacteria
  • Porphyra filament albinism pathogenic bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1: Isolation and purification of pathogenic bacteria

[0019] Albino Porphyra shell filaments from a laver nursery in Lianyungang, Jiangsu in October 2018 ( figure 1 ) to isolate bacteria. Rinse the surface of the diseased shells with sterilized seawater (filtered with glass fiber membrane, heated to boiling, keep boiling for 2 minutes and then cooled for use) for 3 to 2 times, and then wipe dry with sterilized cotton balls (sterilized by moist heat) . Use a sterilized razor blade to scrape the affected part of the shell filament until the white calcareous part is exposed. Dissolve the scrapings with 1mL sterile seawater and fill them into a 1.2mL sterile centrifuge tube. The lysate was then serially diluted to 10 with sterile seawater -2 、10 -3 times.

[0020] Take 100 μL of the above diluted solution and spread them on sterilized TSA medium, 2216E seawater medium and TCBS medium plates with a NaCl concentration of 1%, respectively, and incubate at 28°...

Embodiment 2

[0023] Embodiment 2: the research of bacterial strain pathogenicity

[0024] The GN-R-1 strain was inoculated in 2216E liquid medium at a ratio of 1% (1:100), 180r / min, and cultured at 28°C for 16h. Collect the bacterial solution in a sterile centrifuge tube and centrifuge at 8000 r / min for 2 min. After centrifugation, discard the supernatant and collect the pellet. Resuspend and wash with an appropriate amount of sterilized nutrient seawater, and repeat 3 times. After resuspension and washing, add appropriate amount of sterilized nutrient seawater to resuspend to make bacterial suspension. The bacterial concentration was calculated by the plate colony count method. Adjust the bacterial concentration to 10 7 About CFU / ml.

[0025] The free filaments of Porphyra used for infection were provided by the algae bank of the Key Laboratory of Marine Biotechnology in Zhejiang Province. Use sterile seawater to cultivate filaments, add Ningda No. 3 mother liquor, and the nutrient ...

Embodiment 3

[0029] Example 3: 16S rRNA coding sequence analysis and strain characteristics of GN-R-1

[0030] Bacteria genomic DNA is extracted, with the following primers (sequence information is as follows:

[0031] 27F: 2'-GAGTTTGATCCTGGCTCAG-3';

[0032] 1429R: 2'-GGTTACCTTGTTACGACTT-3') for PCR amplification of 16S rDNA. After NCBI and phylogenetic tree comparison ( Figure 4 ), the bacteria were identified as belonging to the genus Loktanella.

[0033] The strain was named Loktanella sp. GN-R-1, and it was deposited in the General Microbiology Center of China Committee for the Collection of Microorganisms, with the preservation number CGMCC No.21174, and the preservation date was November 13, 2020; Preservation address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing.

[0034] The colonies of Loktanella sp. GN-R-1 strain are opaque, raised, and have regular edges. Scanning electron microscopy showed that GN-R-1 was oval and had flagella.

[0035] Set the envir...

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Abstract

The invention provides a pathogenic bacterium of laver filament albinism. The preservation number of the pathogenic bacterium is CGMCC No.21174. The strain provided by the invention is applied to screening of an albinism-resistant laver strain. A free protonema infection experiment proves that the laver protonema albinism pathogenic bacterium, namely, the rokella GN-R-1 strain obtained by screening can cause albinism of healthy laver protonema and is a pathogenic bacterium of albinism in the laver seedling stage; the separation and discovery of the strain are beneficial to porphyra albinism identification and disease prevention and control, and the strain can be used for screening porphyra strains with albinism resistance.

Description

technical field [0001] The invention belongs to the technical field of disease prevention and control of cultured seaweed, and in particular relates to a pathogenic bacterium of laver filamentous albinism. Background technique [0002] Seaweed is an important economical algae, known as the best seafood, delicious and nutritious. Places such as my country's Shandong and Jiangsu are mainly cultivated Porphyra variegata, and places such as Zhejiang and Fujian are mainly cultivated Porphyra laver. At present, my country is the world's largest producer of seaweed, with an annual cultivation area of ​​over one million hectares and an annual output of more than 1.7 million tons. It is an important pillar of my country's seaweed farming industry. With the expansion of breeding scale, the deterioration of breeding environment, the degradation of seedlings and improper breeding management and other problems are becoming more and more serious, the contradiction between the frequent oc...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12Q1/04C12Q1/18C12Q1/02A01G33/00A01G7/06C12R1/01
CPCC12Q1/04C12Q1/18C12Q1/025A01G33/00A01G7/06G01N2333/195Y02A40/80
Inventor 刘棋琴杨锐史含梦王颖骆其君陈娟娟陈海敏
Owner NINGBO UNIV
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