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A special culture medium for isolating insect endophytic bacteria and its preparation method

An endophytic bacteria and culture medium technology, applied in the field of bioengineering, can solve the problems of difficulty in separating rare bacteria or uncommon bacteria resources, unable to achieve the original intention of the experiment, and the lack of good targeting of insect endophytic bacteria. Rich diversity, strong feasibility, and the effect of improving bacterial yield

Inactive Publication Date: 2018-02-23
HENAN UNIV OF SCI & TECH
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

[0005] In the above-mentioned technical scheme, although adopting beef extract peptone medium can isolate insect endophytic bacteria, existing problems are also relatively prominent, and these deficiencies are concentrated in two aspects (1) as the most traditional and commonly used bacterial culture medium formula, After a long period of repeated use, it has been difficult to isolate rare or uncommon bacteria resources, and many times the original intention of the experiment cannot be achieved; (2) Beef extract peptone medium is suitable for the isolation and culture of general bacteria, especially soil samples, active Experimental samples such as sludge and water samples are not well targeted to insect endophytic bacteria

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The medium formula (1000ml) of embodiment 1 insect endophytic bacteria isolation culture:

[0023] Tenebrio molitor dry powder 2g, yeast powder 1g (G0961, Sangon Bioengineering (Shanghai) Co., Ltd.), gellan gum 3g (EZ0001, Shanghai Eka Biotechnology Co., Ltd.), trehalose 0.5g (TB0966, Sangon Bioengineering (Shanghai) Co., Ltd.), carrot juice 100ml (prepared from 300g red carrots), NaCl 9g (analytical pure, Xilong Chemical), agar powder 16g (01-023, Beijing Aoboxing Biotechnology Co., Ltd.), distilled water to volume To 1 000ml, natural pH.

Embodiment 2

[0024] The preparation process of embodiment 2 medium:

[0025] A. Preparation of Tenebrio molitor dry powder: Weigh an appropriate amount of mature larvae of Tenebrio molitor, dry at 50°C for 12 hours, grind with a mortar until there are no obvious particles, and pass through a 200-mesh sieve to obtain Tenebrio molitor dry powder.

[0026] B. Prepare carrot dipping juice: weigh 300g red carrot, cut into 2cm 3 After crushing the cube with a small pulverizer, transfer it to a conical flask, add 200ml of distilled water, 180r / min, shake and mix at room temperature for 20min. Finally, filter with four layers of gauze, and filter the filtrate for a second time with a sand core filter to obtain the required carrot dipping juice, which is made now and placed in an oven at 60°C to keep warm.

[0027] C. First weigh 5g of gellan gum, slowly dissolve it with 800ml of distilled water, then add 2g of Tenebrio molitor dry powder, 1g of yeast powder, 0.5g of trehalose, and 9g of NaCl, and...

Embodiment 3

[0029] Use the prepared medium to make a plate, and use the dilution of the intestinal homogenate of tussah silkworm Antheraea pernyi to coat the plate and culture it. Each sample is repeated 3 times. The obtained typical strains were identified by the combination of characteristics, physiological and biochemical index determination and 16S rRNA sequence alignment. The results show that the strains grow vigorously on the invented culture medium, the number is large, the reproducibility is strong, and the identified species are abundant. The list of main species is shown in Table 1. The endophytic bacteria in tussah silkworm gut reported in the literature mainly belong to Bacillus, Staphylococcus and Enterobacter. Therefore, compared with the traditional formula, the species of endophytic bacteria isolated by the formula of the present invention has obvious advantages in diversity.

[0030] Table 1 The intestinal endophytic bacteria of tussah silkworm isolated and purified by ...

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Abstract

The invention discloses a special-purpose medium for separating insect endophytic bacteria and a preparation method thereof. A formula comprises the following components: 2g tenebrio molitor dried powder, 1g yeast powder, 3g gellan gum, 0.5g mycose, 9g NaCl, 100ml carrot extracting juice and 18g agar powder, the distilled water is added by metering volume to 1000ml, and the natural pH value is 7.0-7.2. According to the invention, the components are carefully selected, the preparation process is easily understood and is conveniently carried out, the prepared medium has good condition, and is suitable for bacteria separation and culture, and can be used for lineation purifying culture.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a special culture medium for isolating insect endophytic bacteria and a preparation method thereof. Background technique [0002] Biological resources are genetic resources, organisms or parts thereof, populations of organisms or any other biological component of an ecosystem that has actual or potential use or value for humans. The continuous expansion and development of biological resources is an inevitable trend and choice for the development of today's society. The development of biological resources derived from microorganisms has always been a hot field of research. Among them, microbial resources derived from insect endogenous sources have increasingly received people's attention, because there are no infinite resources in nature. Fungal animals, as an important part of the animal kingdom, have many types of insects, wide distribution, and rich ecotypes, and have huge...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12R1/01C12R1/385C12R1/085
CPCC12N1/20
Inventor 孙会忠宋月芹王小东董钧锋纠敏
Owner HENAN UNIV OF SCI & TECH
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