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Low-temperature-resistant lactobacillus strain lactobacillus plantarum CCZZ1 and application thereof

A technology of low temperature resistance and Lactobacillus plantarum, applied in the direction of application, bacteria, microorganisms, etc., can solve the problems of decline, poor quality, weak vitality of lactic acid bacteria, etc., and achieve the effect of overcoming poor fermentation, improving fermentation quality, and fast growth

Active Publication Date: 2014-05-28
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Conversely, undesirable microorganisms grow rapidly, leading to silage spoilage or poor quality
Lactic acid bacteria are the key bacteria for silage fermentation. If the number of lactic acid bacteria attached to the material itself is small, or the number of lactic acid bacteria is large, but the quality is not good (the vitality of lactic acid bacteria is weak) or it is not resistant to special environments (such as low temperature), it is difficult to obtain high-quality silage feed
[0004] Adding acidic chemicals such as formic acid during silage can directly reduce pH and improve the fermentation quality of silage, but it is difficult to use in production due to factors such as high cost, strong corrosion, and unsafe operation.
There are also many lactic acid bacteria additives on the market to improve the fermentation quality of silage, but generally do not take low temperature conditions into account, or the improvement effect is not obvious in low temperature environments

Method used

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  • Low-temperature-resistant lactobacillus strain lactobacillus plantarum CCZZ1 and application thereof
  • Low-temperature-resistant lactobacillus strain lactobacillus plantarum CCZZ1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1 screens low-temperature resistant good lactic acid bacteria

[0024] Isolate 90 strains of lactic acid bacteria from Kinggrass, Stylophyllum, corn, wild rice and other materials, carry out Gram staining and cell shape observation, and according to the growth temperature (10, 15, 20, 25, 30, 35, 40, 45 , 50°C) and growth pH (3.5, 3.75, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0) and other experiments screened out low temperature resistant, acid resistant and fast growing lactic acid bacteria CCZZ1. The results showed that the CCZZ1 strain was a Gram-positive, homofermentative bacillus that could grow at 10°C and pH 3.75, had strong resistance to low temperature and acid, and could ferment most sugars (Table 1).

[0025] Table 1 Characteristics of strain CCZZ1

[0026] strain CCZZ1 source corn shape Rod a + Fermentation type the same type Fertility temperature 10 ℃ + 15 ℃ ++ 45 ℃ + ...

Embodiment 2

[0036] The identification of embodiment 2 lactic acid bacteria

[0037] The strains were cultured overnight at 37°C in 5 mL of MRS medium, transferred to a 1.5 mL centrifuge tube, centrifuged at 10,000 rpm / min for 3 to 5 min to collect the bacteria, and washed with TE0.1 (10 mmol / L Tris–HCl, 0.1 mmol / L EDTA, pH 8.0), washed twice, and then extracted DNA with TIANamp Bacteria DNA Kit (TIANGEN BIOTECH CO., LTD, Beijing, China), and detected its absorbance at OD600 nm. Then, carry out PCR amplification, 16S rDNA amplification primers are 25f (SEQ ID NO:1) and 1492r (SEQ ID NO:2), RCR reaction is 95°C (5 min) - 94°C (30 s) - 55 °C (1 min)-72 °C (1.5 min)-72 °C (10 min), 94 °C (30 s)-55 °C (1 min)-72 °C (1.5 min) reaction cycle 30 times. The amplified product was sent to Boshang Biotechnology Co., Ltd. (China) for sequencing, and the results were compared in the NCBI gene bank to find out the standard strain Lactobacillus plantarum IMAD 80188 ( Lactobacillus plantarum IMAD 801...

Embodiment 3

[0038] Embodiment 3 is added to the effect in the silage

[0039] Silage addition experiments were carried out with lightly predried (predried refers to drying to reduce moisture content) and non-predried ryegrass as materials. Cut ryegrass to 1 cm to 2 cm, mix evenly, and add about 1.0×10 strain CCZZ1 per kg of fresh material 8 indivual. The ryegrass feed added with the strain CCZZ1 was packed into 30 cm×20 cm polyethylene silage bags, 3 bags per treatment, about 200 g per bag, and vacuum sealer (SINBO Vacuum Sealer, Hong Tai Home Electrical Appliance Co. Ltd.) was pumped, sealed, and stored at 15°C for 30 days.

[0040] The rice was cut to 1-2 cm, mixed evenly, and about 1.0×108 strain CCZZ1 was added per kg of fresh material. The rice feed added with strain CCZZ1 was put into 30 cm×20 cm polyethylene silage bags, 3 bags for each treatment, each bag was about 200 g, pumped and sealed with a vacuum sealing machine, and placed at room temperature (25°C-30°C). ℃) for 30 ...

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Abstract

The invention discloses a low-temperature-resistant lactobacillus strain which is lactobacillus plantarum CCZZ1, and an application thereof. The lactobacillus strain provided by the invention is collected at China General Microbiological Culture Collection Center with an address of Institute of Microbiology, Chinese Academy of Sciences, No.3, Court 1, West Beichen Road, Chaoyang District, Beijing. The strain has a collection number of CGMCC NO. 6078. A collection date is May 3rd, 2012. The lactobacillus strain provided by the invention is a Gram stain positive bacilli. The lactobacillus strain is glucose-homofermentative, and is acid-resisting. The strain has high growth speed and wide reproductive temperature range. With the strain, under a low-temperature or normal-temperature environment, silage fermentation quality can be substantially improved.

Description

technical field [0001] The invention relates to the field of microbial application and feed preparation and processing, in particular to a strain of low-temperature resistant lactic acid bacteria Lactobacillus plantarum CCZZ1 and its application in silage preparation. Background technique [0002] Silage is a long-term storage of fresh pastures, forage crops and other multi-moisture feeds in anaerobic and acidic environments. In recent years, with the rapid development of my country's dairy industry, the production of silage has been increasing rapidly. In 2003, the production of silage in China has exceeded 200 million tons (China Dairy Industry Association, 2003). It can be seen that the production and utilization of silage is of great significance to animal husbandry production, especially for high-yield dairy cows. It can not only ensure the quantity and quality of milk and meat products, but also save feed grain. In addition, through silage, rough or smelly plant mat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23K1/14C12R1/25
Inventor 张建国陈明霞
Owner SOUTH CHINA AGRI UNIV
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