Microorganisms, methods and kits for promoting the growth, photosynthesis and secretion of ciguatoxin of Gambia algae

A technology for the growth of microorganisms and algae, applied in the biological field, can solve the problems of unsatisfactory demand, limited toxin amount, high price, etc., and achieve the effect of promoting the growth of gambiae algae

Active Publication Date: 2019-07-16
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the source of standard ciguatera toxin is mainly from highly toxic fish in natural toxin-producing areas, but the amount of toxin obtained in this way is very limited and expensive (the conservative market price is about 20,000-35,000 RMB / μg), unable to meet the needs of the global market

Method used

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  • Microorganisms, methods and kits for promoting the growth, photosynthesis and secretion of ciguatoxin of Gambia algae
  • Microorganisms, methods and kits for promoting the growth, photosynthesis and secretion of ciguatoxin of Gambia algae
  • Microorganisms, methods and kits for promoting the growth, photosynthesis and secretion of ciguatoxin of Gambia algae

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] The isolation and identification of embodiment 1 bacterial strain

[0082] 1. Isolation of strains

[0083] Four sampling points with different toxicity in Marakei Atoll (Marakei), Republic of Kiribati were selected for sampling, including two strong cigara toxicity areas M1 (2°01.394'N, 173°15.383'E) and M2 (1°59.879' N, 173°15.032'E), and two weak ciguatera toxicity regions M3 (1°58.246'N, 173°16.268'E) and M4 (2°00.150'N, 173°18.010'E). Collect samples from macroalgae, corals, seaweeds and other areas where gambiae are easy to attach, for algae isolation and bacterial screening. The sampled samples were aseptically preserved and brought back to the laboratory (Marine Ecology Laboratory, Shenzhen Graduate School, Tsinghua University) aseptically spread on a 12cm medium-sized solid medium plate containing 2216E, and cultured overnight at 30°C until clear Visible monoclonal.

[0084] Then about 200 single clones were picked from the culture plate and placed in 2ml EP...

Embodiment 2

[0099] Embodiment 2, Bacillus thuringiensis serovar konkukian (Bacillus thuringiensis serovar konkukian) GDMCCNo.60074 promotes the detection of gambiae growth and photosynthesis

[0100] 1. Preparation of experimental algal species

[0101] According to the source and toxicity of the isolated algal species of Gambiae gambiae, the strong ciguatoxin-producing sp. gambiae M2 was selected as the experimental object to study the growth-promoting effect of Bacillus thuringiensis serovar konkukian (Bacillus thuringiensis serovar konkukian) GDMCC No.60074. .

[0102] Algae cultivation: use a 40X optical microscope to observe the growth of algae gambiae in each well of the 96-well plate. Transfer the successfully propagated and well-grown Algae gambiae to 10ml of fresh K medium for expansion. After 10 days of culture, transfer 5ml of Gambiae to Corning 25cm containing 20ml of fresh K medium 2 The expanded culture was carried out in air-permeable cell culture flasks, and the concent...

Embodiment 3

[0139] Embodiment 3, Bacillus thuringiensis serovar konkukian (Bacillus thuringiensis serovar konkukian) GDMCCNo.60074 increases the detection of ciguatoxin secretion

[0140] 1. Extraction of ciguatera toxin

[0141] 1) Use a 0.22 μm water-phase microporous filter membrane to filter the algae liquid, and transfer the filter membrane to a 50ml centrifuge tube.

[0142] 2) Add 25ml of anhydrous methanol to the centrifuge tube and shake to fully immerse the filter membrane.

[0143] 3) Place the centrifuge tube in an ultrasonic wall breaker under ice bath conditions, and pre-rinse the probe.

[0144] 4) Set the Pulse to be on for 5 seconds, off for 1 second, power at 40%, and each tube to be sonicated for 5 minutes.

[0145] 5) Transfer the sonicated sample to an ultracentrifuge tube, and centrifuge at 12000 rpm for 10 minutes.

[0146] 6) Transfer the supernatant to a 250ml round-bottomed flask, add 25ml of anhydrous methanol to the centrifuge tube and pipette the precipitat...

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Abstract

The invention provides a microorganism, a method and a kit for promoting the growth, photosynthesis and secretion of ciguatoxin of Gambiae algae. The microorganism is Bacillus thuringiensis sp., which was deposited in the Guangdong Provincial Microorganism Culture Collection Center on September 5, 2016, and the deposit number is GDMCC No.60074. The microorganism of the invention can not only increase the biomass and photosynthetic efficiency of algae gambiae, but also increase the secretion of ciguatoxin, obtain a new method for the source of ciguatoxin standard, and contribute to the fine research of the toxin.

Description

technical field [0001] The present invention relates to the field of biology. Specifically, the present invention relates to microorganisms, methods and kits for promoting the growth, photosynthesis and secretion of ciguatoxin of algae gambiae. Background technique [0002] Ciguatera fish poisoning (CFP), also known as ciguatera toxin, is an algae toxin that seriously endangers human health, mainly produced by benthic Gambierdiscus sp. The toxin has a food chain cumulative effect. The toxic gambiae is enriched in the fish after being ingested by fish, and the fish meat flows through the food chain to the dining table and finally threatens human health. With the expansion of the scale of global fishery trade, poisoning incidents are increasing year by year. Consumption of ciguatera toxin can easily lead to abnormal functions of the nervous system, cardiovascular system and gastrointestinal system, which is extremely harmful to human health. In addition, ciguatera toxin is f...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P1/06C12N1/38C12N1/12C12P17/18C12R1/07
CPCC12N1/12C12N1/38C12P1/06C12P17/181C12N1/205C12R2001/075
Inventor 蔡中华周进王博兰霞劳永民晋慧姚蜜蜜
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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