A pathogenic bacterium of kelp albinism

A technology for albinism and pathogenic bacteria, applied in the directions of bacteria, biochemical equipment and methods, microorganisms, etc., can solve the problem of difficult to determine the pathogenic bacteria of kelp disease

Active Publication Date: 2021-04-09
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, it has been difficult to identify the causative agent of kelp disease

Method used

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  • A pathogenic bacterium of kelp albinism
  • A pathogenic bacterium of kelp albinism
  • A pathogenic bacterium of kelp albinism

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Embodiment 1: Isolation and purification of pathogenic bacteria

[0014] In December 2017, bacteria were isolated from albino kelp seedlings in a kelp nursery in Rongcheng, Shandong. Select the kelp seedlings with albinism, wash away the surface impurities with sterile seawater, and grind them into a homogenous slurry in a sterile mortar. Dilute the homogenate by 10 with sterile seawater -2 、10 -3 、10 -4 、10 -5 For gradient dilution, take a little (0.5mL) of different dilutions and spread them on Zobell 2216E solid medium with a triangular sterile glass coating rod. Cultivate in a constant temperature culture at 25°C for 24-72h, and observe the growth and shape of the colonies. Then use the inoculation loop to pick 10 -3 The colonies on the dilution gradient plate were further separated and streaked on Zobell 2216E solid medium, and purified until a single colony was isolated. In this purification experiment, a total of 10 purified strains were isolated, and it w...

Embodiment 2

[0015] Embodiment 2: the research of bacterial strain pathogenicity

[0016] Take out the preserved bacterial liquid from the -20°C refrigerator, absorb 1 mL into 100 mL Zobell 2216E liquid medium and cultivate in a shaking incubator. After 24 hours, absorb 1 mL into 100 mL Zobell 2216E liquid medium to continue culturing. After 12 hours, absorb 2 mL of overnight culture solution Continue to culture in 200mL Zobell 2216E liquid medium, the culture condition is 25°C, and the rotation speed is 120r / min. After 4 hours, the bacterial liquid was drawn to measure the concentration of the bacterial liquid. At this time, the growth of the bacterial solution is in the logarithmic growth phase.

[0017] Select healthy kelp seedlings in good growth state, place them in a petri dish filled with sterile sea water, and then cut the sporophytes into small pieces (1.0cmx1.0cm) with a sterilized blade. Put the kelp tissue block into the culture plate, and add 1mL X-8 bacterial solution (conc...

Embodiment 3

[0019] Embodiment 3: Analysis of pathogenic strains

[0020] Extract bacterial genomic DNA, the specific primer sequence information is as follows:

[0021] 27F: 5'-GAGTTTGATCCTGGCTCAG-3'; 1429R: 5'-GGTTACCTTGTTACGACTT-3' PCR amplification of 16S rDNA, the size of the amplified fragment is 1397bp ( figure 2 ).

[0022] After NCBI comparison, the screened strain X-8 had the highest similarity with two strains of piscicidal Pseudoalteromonas bacteria from the coast of Delaware, Pesudoalteromonas piscicida strain DE2-A and Pesudoalteromonas piscicida strain DE2-B; Therefore, it is named as Pseudoalteromonas X-8 strain (Pesudoalteromonas X-8). 17th; Preservation address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing.

[0023] The X-8 colonies are orange in color, opaque, raised, and have regular edges. Scanning electron microscopy revealed that X-8 is rod-shaped with polar single flagella ( image 3 ).

[0024] The culture conditions of the X-8 strain are...

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Abstract

The invention provides a pathogenic bacterium of kelp albinism, which is Pseudoalteromonas X-8 strain (Pesudoalteromonas X-8), which is preserved in the General Microbiology Center of China Microbiological Culture Collection Management Committee, with a preservation number of CGMCC20099 and a preservation date of June 17, 2020; Preservation address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. The seawater Pseudoalteromonas X-8 strain provided by the present invention can be used to screen albinism-resistant kelp strains. The Pseudoalteromonas X-8 strain obtained by screening of the present invention is confirmed by restaining healthy kelp seedlings, which can cause healthy kelp to produce albinism, and is a pathogenic bacterium of albinism in kelp seedling stage; use this bacterial strain to be used for screening. Albino kelp strains.

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 kelp albinism pathogenic bacterium. Background technique [0002] Kelp is an important large-scale economically cultivated seaweed in my country, with an annual output of about 1.5 million tons. Kelp culture has become one of the main pillar industries of my country's mariculture industry. Like higher crops, kelp also has diseases in the seedling and breeding stages, which can lead to a 20-50% yield reduction in severe cases. Because algae pathogenic bacteria are mostly conditional pathogens, that is, under normal circumstances, they exist on healthy algae and do not show pathogenicity. Only when environmental factors change (such as temperature rise, light changes, and environmental pollution, etc.) showed pathogenicity. With the development of the kelp aquaculture industry, the frequent outbreaks of diseases in the pro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12Q1/18C12R1/01
CPCC12Q1/18C12N1/205C12R2001/01
Inventor 王高歌张晓阳韩琦
Owner OCEAN UNIV OF CHINA
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