A method for preparing bacteria and algae cells by two rounds of co-cultivation and flocculation

A technology of co-cultivation and charcoal algae is applied in the field of preparing bacteria and algae cells by two rounds of co-cultivation and flocculation, which can solve problems such as increasing the dry weight of plants, improve growth rate and biomass, reduce production and development costs, and reduce Effects of industrial production and development application costs

Inactive Publication Date: 2019-03-26
ZHEJIANG GONGSHANG UNIVERSITY
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Problems solved by technology

Rice plants contain many endophytic microorganisms, which are important endophytic bacterial resources, among which Pantoea is a common endophytic bacterium in rice, which can promote plant growth and increase plant dry weight [Sánchez-Matamoros R C, etal.Structure of the O-antigen of the lipopolysaccharide isolated from Pantoeaananatis AEP17, a rhizobacterium associated with rice.Carbohydrate Research, 2013,369:25-30.], the main mechanism of Pantoea endophytica in rice to promote the growth of host rice includes secreting auxin, promoting nitrogen fixation and dissolving phosphorus, etc. (RuizaD, et al.. Characterization and screening of plant probiotic traits of bacteria isolated from rice seeds cultivated in Argentina. The Journal of Microbiology, 2011, 49(6): 902-912), because microalgae have both plant cell photosynthesis Theoretically, endophytes that have symbiotic functions on plants may also play a similar biological role on microalgae cells, and a small amount of research evidence has shown that microorganisms that have growth-promoting effects on plants can also Improve the biomass of microalgae (Kim B H, et al.Role of rhizobium, a plant growth promotingbacterium, in enhancing algal biomass through mutualistic interaction. Biomass and Bioenergy, 2014, 69(3): 95-105.), but so far at home and abroad There is no research on the development of new technologies for the production of chlorella using the symbiosis of rice endophytes and microalgae

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  • A method for preparing bacteria and algae cells by two rounds of co-cultivation and flocculation

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Embodiment 1

[0029] 1) The rice endophytic Pantoea strains were purchased from the China Agricultural Microorganism Collection Center (ACCC), coded as 10454. After the strains were activated by conventional methods, they were inoculated into a Erlenmeyer flask containing 10 mL of LB liquid medium, and carried out For expansion culture, the bacteria liquid in the logarithmic growth phase after 16 hours of expansion culture is inoculated according to the volume ratio of bacterial liquid: liquid LB medium = 1:10, and then the second expansion culture is carried out. The culture conditions are 37°C, 200rpm constant temperature shaker Cultivate in the dark. The second expansion culture 10h to OD 600 =1 is used as the seed cell of the co-cultivation of bacteria and algae, according to the growth curve, the bacterial growth is in the logarithmic phase, and the effective number of viable bacteria (CFU, Colony-Forming Units) in the culture solution is measured by the conventional plate counting met...

Embodiment 2

[0035] 1) The rice endophytic Pantoea strains were purchased from the China Agricultural Microorganism Collection Center (ACCC), coded as 10454. After the strains were activated by conventional methods, they were inoculated into a Erlenmeyer flask containing 10 mL of LB liquid medium, and carried out For expansion culture, the bacteria liquid in the logarithmic growth phase after 16 hours of expansion culture is inoculated according to the volume ratio of bacterial liquid: liquid LB medium = 1:10, and then the second expansion culture is carried out. The culture conditions are 37°C, 200rpm constant temperature shaker Cultivate in the dark. The second expansion culture 10h to OD 600 =1 is used as the seed cell of the co-cultivation of bacteria and algae, according to the growth curve, the bacterial growth is in the logarithmic phase, and the effective number of viable bacteria (CFU, Colony-Forming Units) in the culture solution is measured by the conventional plate counting met...

Embodiment 3

[0041] 1) The rice endophytic Pantoea strains were purchased from the China Agricultural Microorganism Collection Center (ACCC), coded as 10454. After the strains were activated by conventional methods, they were inoculated into a Erlenmeyer flask containing 10 mL of LB liquid medium, and carried out For expansion culture, the bacteria liquid in the logarithmic growth phase after 16 hours of expansion culture is inoculated according to the volume ratio of bacterial liquid: liquid LB medium = 1:10, and then the second expansion culture is carried out. The culture conditions are 37°C, 200rpm constant temperature shaker Cultivate in the dark. Continuous expansion was carried out three times, and the culture conditions were 37°C, 200rpm constant temperature shaker and dark culture. The third expansion culture 10h to OD 600 =1 is used as the seed cell of the co-cultivation of bacteria and algae, according to the growth curve, the bacterial growth is in the logarithmic phase, and t...

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Abstract

The invention discloses a method for preparing bacterium-algae cells through a two-wheel coculturing and flocculation method. The method comprises the steps of 1, conducting the first cycle of coculturing on rice endogenous pantoea which is added for the first time and chlorella under illumination, so that after culturing is ended, a bacterium-algae coculturing solution of the first cycle is obtained; 2, supplementing rice endogenous pantoea which is added for the second time into the bacterium-algae coculturing solution of the first cycle, conducting the second cycle of bacterium-algae coculturing under illumination, so that after culturing is ended, a bacterium-algae coculturing solution of the second cycle is obtained; 3, conducting flocculation collection treatment on the bacterium-algae coculturing solution of the second cycle, so that the bacterium-algae cells are obtained. According to the method, bacterium-algae coculturing is conducted through the rice endogenous pantoea and the chlorella, the growth rate of the algae cells is raised, biomass of the algae cells is increased, meanwhile, flocculation performance of the algae cells is improved, application of chemical flocculating agents and toxic harm to the algae cells are reduced, algae cell collection efficiency is improved, algae cell collection cost is lowered, and thereby industrial production and development and application cost of chlorella are remarkably lowered.

Description

technical field [0001] The invention relates to the technical field of co-cultivation of bacteria and algae, in particular to a method for preparing bacteria and algae cells by two rounds of co-cultivation and flocculation. Background technique [0002] Chlorella is a single-celled green algae, which has strong environmental adaptability and is easy to cultivate on a large scale. The cells contain rich nutrients and are widely used in health food, feed, food additives, finely processed products and as raw materials for pharmaceutical preparations . Chlorella is rich in short-chain fatty acids and is a high-quality biodiesel raw material (Xu Jin, Xu Xudong, Fang Xiantao, Hu Hanhua. Screening of high oil-yielding chlorella and oil analysis. Acta Hydrobiology, 2012,36(3):426 -432.). Chlorella pyrenoidosa is a common chlorella species in my country. It has high protein content and rich essential amino acids. It is an excellent protein source for feed (Li Guoping. Analysis of a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/12C12R1/89
CPCC12N1/12C12N1/20
Inventor 赵艳
Owner ZHEJIANG GONGSHANG UNIVERSITY
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