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A kind of Paenibacillus aerobacillus and its application
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A technology of bacillus and strains, applied in the field of microorganisms, can solve problems such as reduced production efficiency, and achieve the effect of good anti-penetration ability
Active Publication Date: 2021-12-24
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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Problems solved by technology
At present, Bacillus licheniformis and Bacillus subtilis are the most commonly used in fermentation engineering, but as the fermentation time prolongs, their production efficiency will also decrease.
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Embodiment 1
[0043] For the long-term paddy rice cultivation from Yanji City of Jilin Province in the rice rhizosphere soil collected examples used in this example.
[0044] Soil samples were collected, weighed 0.2g soil samples in 20mL R 2 A liquid medium at 30 ℃, 220rpm the enrichment culture 3d, taking a first enrichment culture 200μL, was added to 20mL fresh R 2 A broth culture 3d, the above-described operation is repeated twice, to give a final draw enrichment cultures 100 uL, 10 in accordance with -1 10 -2 10 -3 10-4 10 -5 And 10 -6 Sequentially dilutions were applied to take 200μL R 2 On A agar medium plates 30 upside down in an incubator culturing deg.] C 3d, isolates obtained.
[0045] The isolates were visible, rod-shaped bacteria, no flagella under a transmission electron microscope (e.g. figure 1 Below); this strain R 2 A colony on the agar medium showed yellow, convex shape, surface wetting, neat edge, no extracellular secretion (e.g. figure 2 Indicated.
[0046] Physiological and...
Embodiment 2
[0049] Example aerobic classes embodiment the isolated Bacillus genome using R14 Company bacterial genomic DNA extraction kit (centrifugal bars) extracted. Aerobic Paenibacillus 16S rDNA R14 PCR amplified with the universal primer sequences designed reference William G.Weisburg (1991) article, the primers are synthesized by gene technology Ltd. Beijing Genomics Luk. Amplification kit used to buy Nanjing Wei Chan Connaught Biotechnology Limited.
[0050] Positive primer
[0051] 27F (5'-AGAGTTTGATCATGGCTCAG-3 ', SEQ ID NO.4),
[0052] Reverse primer
[0053] 1492R (5'-TACGGTTACCTTGTTACGACTT-3 ', SEQ ID NO.5).
[0054] Table 1 shows the PCR amplification system:
[0055] Element Volume / UL 2 × Phanta PCR MIX BUFFER 25 27F 2 1492R 2 R14 genome 2 Sterilization DDH 2 O
Position to 50 ul
[0056] Table 2 PCR reaction procedure:
[0057]
[0058] The amplified 16S rDNA sequence is connected to The company's pJET 1.2 / blunt carrier, commissio...
Embodiment 3
[0060] Example 1 Class aerobic isolated Bacillus embodiment will be obtained on LB plates scribe line activation R14, 30 ℃ inverted culture. Colonies on the plates were picked and inoculated into 1 / 3TSB liquid culture medium, 30 ℃, 220rpm, bacteria shaking overnight. Determination of OD 600 , When OD 600 = 0.1, added to fresh LB broth, 30 ℃, 220rpm, grown to OD 600 0.4 to 0.5, bacteria take 100μL 900μL PBS was added to 2mL EP bacteria as a sample tube stand.
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Abstract
The invention provides a Paenibacillus aerobacillus, the classification of the Paenibacillus aerobic is Paenibacillus aerobic, and the preservation number of the Paenibacillus aerobic is GDMCC NO: 60620; the 16S rDNA sequence of the Paenibacillus aerobic As shown in SEQ ID NO:1. The Paenibacillus aerobacillus R14 has good oxidation resistance and can be used as an antioxidant strain; it can also participate in the construction of transgenic organisms with antioxidant functions; and it has potential value in mutation breeding and improving the antioxidant functional traits of plants ; At the same time, the strain has good anti-osmosis ability, and can participate in the construction of engineering strains with anti-osmosis function.
Description
Technical field [0001] The present application relates to the field of microbial, particularly, to an aerobic Paenibacillus their applications. Background technique [0002] Suitable environmental conditions is a prerequisite for good plant growth, the plants are abiotic stress (oxidative, high permeability, etc.), its growth will be inhibited, under conditions of oxidative stress will cause greatly increased concentration of free radicals. Wherein the oxidative stress is generated reactive oxygen species (ROS) of genomic DNA and biomacromolecules (especially proteins) in the cells will cause severe damage, and the bacteria to adapt to reactive oxygen species (ROS) by increasing the detoxifying enzyme and repair proteins expressed increases. Screening strains have antioxidant properties, identify the relevant functional genes, and then transformed into plants, can help to cultivate plants with resistance to adapt to the external environment changing climate it has important pract...
Claims
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