Pear branch degradation bacteria L2 and inoculant thereof

A technology for bacterial inoculum and pear tree is applied in the field of agricultural waste pear tree pruning branches degrading bacteria L2 and its inoculum, which can solve the problems of air pollution, inability to return to the field, diseases and insect pests, etc., achieve broad application prospects, and promote agricultural The effect of sustainable development and environmental protection

Active Publication Date: 2017-07-21
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, pear tree branches contain a large amount of lignin and cellulose, so they are not easy to rot and cannot be returned to the field in situ.
Except for some branches used...

Method used

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  • Pear branch degradation bacteria L2 and inoculant thereof
  • Pear branch degradation bacteria L2 and inoculant thereof
  • Pear branch degradation bacteria L2 and inoculant thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Enrichment, isolation, screening and identification of pear branch degrading bacteria L2

[0027] 1) Enrichment of highly efficient degrading bacteria from pear tree branches: samples of degrading bacteria were collected from rotten pear tree branches piled up all year round in Lanzhou, and passed through a 20-mesh sieve to make samples. Take 0.2 g of branch samples and mix them in 20 g of solid-state enrichment medium with pear branches as the only carbon source for solid-state enrichment for 30 days, pick 0.2 g of medium with black degradation spots on the surface debris of branches, mix and inoculate into new Continue to enrich for 30d in the solid-state enrichment medium. Continue to pick 1g of the medium with black degradation spots on the surface of the branches and inoculate it into 100mL liquid enrichment medium, culture on a shaker at 160rpm, and add sterile water regularly; after 30 days, inoculate 1mL into the new liquid enrichment medium , continu...

Embodiment 2

[0032] Example 2 Degradation Enzyme Activity Determination

[0033] Inoculate the strain suspension of bacteria L2 by 1% inoculum amount (10 7 / mL) into a Erlenmeyer flask (250 mL) filled with 100 mL of liquid enzyme-producing medium, and placed at 28° C. for 21 d with shaking at 160 r / min. Samples were taken every 2 days, and the fermentation broth was centrifuged to remove bacteria and impurities to obtain a crude enzyme solution, in which the activities of lignin peroxidase (Lip) and manganese peroxidase (MnP) were determined. The result is as image 3 with Figure 4 As shown, the strain L2 can produce lignin-degrading enzymes with strong ability, fast enzyme production, and long-term enzyme production. at 10U·mL -1 The above is more than 3D, which has a good effect of producing Lip. The highest activity of lignin peroxidase is 21.76U·mL -1 , the highest activity of manganese peroxidase was 106.36U·mL -1 .

[0034] The enzyme activity of Lip is defined as the oxidat...

Embodiment 3

[0039] Example 3 Determination of Solid State Degradability

[0040] Prepare the L2 bacteria agent, the method is as follows: (1), inoculate the bacteria L2 in the LB liquid medium, shake at 30°C for 36h, so that the bacteria content reaches 10 9 / mL, the spore formation rate reaches more than 90%; (2), the bacterial suspension 6000rpm that step (1) has cultivated is centrifuged 10min, resuspended with a small amount of LB liquid culture medium, makes bacterial L2 reach 10 10 / mL, stored at 4°C for later use.

[0041] Add 5g of dried pear branch powder (40-60 mesh) and 10mL solid-state culture nutrient solution (solid-state culture nutrient solution: NH 4 Cl 2.0g, MgSO 4 ·7H 2 O 0.5g, KH 2 PO 4 1.0g, Na 2 HPO 4 0.2g, MnSO 4 0.035g, CuSO 4 ·5H 2 O 0.007g, FeSO 4 ·7H 2 (0.007g, water 1L), until the solid and liquid are mixed evenly and flat, sterilized at 121°C for 20min, inoculated with L2 bacterial agent (10 10 / mL), placed in a 30°C incubator for 30 days. Dur...

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Abstract

The invention discloses a bacterium L2 used for degrading pear branches, with a classification name of Bacillus megaterium, preserved in China General Microbiological Culture Collection Center on 8th, July, 2013, with a culture preservation number of CGMCC NO.7899. The bacterium L2 can grow on a culture medium using analogue of lignin, guaiacol as a unique carbon source or pear branch powder as the unique carbon source, and can generate a fading ring on an LB-aniline blue plate. Experiments show that after culture suspension of the acterium L2 with an inoculation amount of 1% is inoculated on the culture medium, and solid state fermentation is performed for 30d, a weight loss ratio of the culture medium can reach 105. During a liquid state fermentation process on the 21st day, the highest value of activity of generated lignin peroxidase is 21.76U*mL-1, and the highest value of activity of generated manganese peroxidase is 106.36U*mL-1.

Description

technical field [0001] The invention belongs to the agricultural intensive production technology, and relates to an agricultural waste pruned branch degrading bacterium L2 and a bacterial agent thereof. The bacterium L2 is specially used for degrading the discarded pear tree branch to produce organic fertilizer, and realizes resource utilization of organic waste. Background technique [0002] Pruning is an important technical measure for the cultivation and management of pear trees. Appropriate pruning has a positive impact on the growth of fruit trees and the yield and quality of fruits. For pear orchards in full fruit stage, the amount of pruned branches is generally 1500-2250kg·hm -2 . Statistics from the Department of Market and Economic Information of the Ministry of Agriculture show that in 2011, the cultivated area of ​​pear trees in my country was more than 1.1 million hectares. According to this figure, the annual pruning of pear orchards in my country can reach 1.6...

Claims

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Application Information

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IPC IPC(8): C12N1/20C05F11/00C05F17/00C12R1/11
CPCC05F11/00C05F17/00C12N1/20C12N1/205C12R2001/11C05F11/08Y02W30/40
Inventor 董彩霞张乃文徐阳春
Owner NANJING AGRICULTURAL UNIVERSITY
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