Microalgae symbiotic bacteria isolation medium and separation method, and high-throughput screening method for key bacteria affecting growth of microalgae

A technology for separating medium and screening method, applied in the field of microalgae commensal bacteria separation, can solve the problems of low screening efficiency and large demand for human and material resources, and achieve the effects of reducing culture temperature, prolonging culture time, and increasing species.

Active Publication Date: 2019-05-07
XIAMEN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the wide variety of algae bacteria, this method requires a lot of manpower and material resources, and the screening efficiency is low.

Method used

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  • Microalgae symbiotic bacteria isolation medium and separation method, and high-throughput screening method for key bacteria affecting growth of microalgae
  • Microalgae symbiotic bacteria isolation medium and separation method, and high-throughput screening method for key bacteria affecting growth of microalgae
  • Microalgae symbiotic bacteria isolation medium and separation method, and high-throughput screening method for key bacteria affecting growth of microalgae

Examples

Experimental program
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Effect test

Embodiment 1

[0077] A method for isolating microalgae symbiotic bacteria in a marine microalgae culture solution, the steps are as follows:

[0078] 1. Medium

[0079] The solid medium of the present invention: 2.5g of peptone, 0.5g of yeast extract, 0.005g of ferric citrate, 1g of methylamine hydrochloride, 1L of artificial seawater, 15g of agar, after stirring evenly, sterilize under high pressure at 121°C for 20 minutes. When cooling to about 60°C, add 1ml / L of f / 2 mother solution, mix carefully to avoid a large amount of air bubbles, pour on a plate, and set aside after solidification.

[0080] 2216E solid medium: 37.4g of commercial 2216E powder, 15g of agar, 1000ml of distilled water, stirred evenly, sterilized at 121°C for 20 minutes, cooled to 60°C, poured onto a plate, solidified and dried before use (as a control).

[0081]Take 1 mL microalgae culture solution from the culture solution of 4 strains of microalgae Porphyridium cruentum CCMA-143, Phaeodactylum tricornutum CCMA-267,...

Embodiment 2

[0089] A high-throughput rapid screening method for key bacteria that affects the growth of Nannochloropsis gaditana CCMP526, specifically comprising the following steps:

[0090] Randomly select 16 strains of bacteria from the algae bacteria library established in Example 1, and screen the bacteria that have a key influence on the growth of the oil-producing model microalgae Nannochloropsis gaditana CCMP526. The specific operations are as follows:

[0091] (1) Determine the scope of microalgae inoculation: 10 9 2-fold serial dilution of Nannochloropsis stock solution of cells / ml until 10 3 Cells / ml, confirmed by counting Nannochloropsis cells of different concentrations by flow cytometry, and using a microplate reader to detect the corresponding chlorophyll fluorescence values ​​of algae cells at each concentration with an excitation light of 488nm and an emission light of 680nm, each The concentration was repeated three times, and the standard curve of the linear relationsh...

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Abstract

The invention relates to the field of microalgae symbiotic bacteria isolation, specifically speaking, relates to a microalgae symbiotic bacteria isolation medium and separation method, and a high-throughput screening method for key bacteria affecting the growth of microalgae. Improvement can be performed based on a common marine bacterial isolation medium - Zobell 2216E marine broth bacterial culture medium, so that the culture medium for increasing the culturable bacterial species of microalgae phycosphere symbiotic bacteria can be provided, and abundant strain resources can be provided for the screening of key bacteria affecting the growth of microalgae. Through the high-throughput rapid screening method for significantly promoting or inhibiting algae growth bacteria, bacteria having keyinfluences on the growth of the microalgae can be discovered in a short time.

Description

technical field [0001] The invention relates to the field of separation of microalgae symbiotic bacteria, in particular to a microalgae symbiotic bacteria isolation medium, an isolation method and a high-throughput screening method for key bacteria affecting the growth of microalgae. Background technique [0002] In the natural water environment and the applied cultivation process of microalgae, there is a close relationship between algae and bacteria, and some bacteria have a key influence on the growth of algae. In 1972, microalgae biologists Bell and Mitchell proposed the concept of Phycosphere (Bell & Mitchell, 1972). Similar to the influence of plant rhizosphere microorganisms on plant growth, the phycosphere often inhabits special groups of bacteria, which play an important role in regulating the growth and physiological morphology of microalgae. In the in-situ study of marine ecology and the co-culture study of algae and bacteria in the laboratory, it was found that ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/02
Inventor 周涵韬李妍妍林艺华杨淋
Owner XIAMEN UNIV
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