Three-year-old peony rhizosphere bacillus megaterium FD3-13 for oil and application thereof
A technology of Bacillus megaterium and peony root, applied in the field of microorganisms, to achieve good results
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Embodiment 1
[0079] Example 1 Isolation and purification of a three-year-old oil strain of Bacillus megaterium FD3-13 from the peony rhizosphere
[0080] Firstly, remove the visible debris on the rhizosphere soil surface of the three-year-old oil peony planted in the experimental farm of Kaiyuan Campus of Henan University of Science and Technology in Luoyang City, Henan Province. According to the five-point sampling method, the rhizosphere soil was collected with a soil drill at a distance of 10 cm from the plant. Put it into a sterile bag and store it in an ice box. After arriving at the laboratory, use a brush to brush off the soil attached to the root system, weigh 5g of fresh soil samples in the ultra-clean workbench into a sterile centrifuge tube, dilute to 50mL with sterile water, shake for 20min, and press 10 The dilutions were carried out sequentially, and a pipette was used to draw 10 -6 ~10 -4 Each 0.1mL sample of the three dilutions was evenly spread on the organophosphate sol...
Embodiment 2
[0082] Example 2 Identification of Moutan Rhizosphere Bacillus megaterium FD3-13 for three-year oil production
[0083] Bacterial genomic DNA was extracted using the AxyPrep Bacterial Genomic DNA Miniprep Kit. Genomic DNA was amplified using bacterial universal primers 27F: 5'-AGAGTTTGATCCTGGCTCAG-3' and 1492R: 5'-CTACGGCTACCTTGTTACGA-3' to obtain PCR products. PCR products were purified with AxyPrep DNA. The purified PCR products were bidirectionally sequenced using the ABI3730-XL sequencer at Shanghai Passino Biotechnology Co., Ltd. After using ChromasPro software to truncate the invalid sequence, use BioEdit software to splice the double-ended effective sequence, and then use the Blast program in the NCBI database to compare the effective sequence with the reference sequence in the NCBI database to obtain a sequence similar to the strain to be tested. The most sexual strain is Bacillus megaterium, with a homology of 99%.
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