An endophytic fungus bdf09 of salvia miltiorrhiza and its application
A BDF09, strain technology, applied in fungi, plant/algae/fungus/moss components, microorganisms, etc., can solve problems that have not yet been discovered, and achieve good anticoagulant effect.
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[0034] (1) The endophytic fungus BDF09 of Salvia miltiorrhiza according to the present invention is isolated from the root of the medicinal plant Salvia miltiorrhiza Bge.f.alba through strict surface disinfection.
[0035] (2) The endophytic fungus BDF09 of salvia miltiorrhiza according to the present invention has a white-purple velvet-like surface on the PDA medium and a purple backside, and grows rapidly. Optical microscope examination showed that the mycelium was colorless, and the microconidia were unicellular or twin-celled macroconidiospores with a little curvature, which was preliminarily determined to be Fusarium sp.
[0036] (3) For the endophytic fungus BDF09 of Salvia miltiorrhiza according to the present invention, its genomic DNA extraction method adopts the improved SDS method, utilizes ITS universal primers ITS1 and ITS4, and performs ITS amplification analysis to obtain a single DNA band with a length of 515bp, submit Blast comparison was performed in the GenB...
Embodiment 1
[0038] Embodiment one: the isolation and screening of endophytic Fusarium BDF09 of salvia miltiorrhiza
[0039] Healthy and disease-free whole plants of Salvia miltiorrhiza Bge.f.alba were collected, rotted leaves were removed, sediment and other impurities were washed with clean water, rinsed, and placed on filter paper to air dry naturally. Cut the root of Salvia miltiorrhiza into small pieces of about 0.5 cm, and place them in a clean Petri dish. In the ultra-clean workbench, treat with 75% ethanol for 2 minutes, rinse with sterile water for 5 times, treat with 0.1% mercuric chloride for 5 minutes, rinse with sterile water for 5 times, and finally rinse with 75% alcohol for 30 seconds, rinse with sterile water for 5 blot the excess water with sterilized filter paper. After peeling off the outer skin of the material with a sterile razor blade, cut into small pieces of 0.2cm×0.2cm, place them in PDA medium supplemented with penicillin, and incubate at a constant temperature ...
Embodiment 2
[0040] Example 2: Identification of Endophytic Fusarium BDF09 in Salvia miltiorrhiza
[0041] (1) Morphological identification:
[0042] The surface of BDF09 colony on PDA medium was white-purple velvet, the back was purple, and it grew rapidly ( figure 1 ). Optical microscope examination showed that the mycelium was colorless, and the microconidia were unicellular or twin-celled macroconidia with a little curvature, which was preliminarily determined to be Fusarium sp. ( figure 2 ).
[0043] (2) Molecular identification:
[0044] 采用改良SDS法提取基因组DNA;利用ITS通用引物ITS1和ITS4,通过PCR扩增其ITS序列,得到长度为515bp的单一DNA条带,其ITS rDNA序列为: TGGGATAGGACTCACTCCCAACCCCTGTGACATACCACTTGTTGCCTCGGCGGATCAGC CCGCTCCCGGTAAAACGGGACGGCCCGCCAGAGGACCCCTAAACTCTGTTTCTATATGT AACTTCTGAGTAAAACCATAAATAAATCAAAACTTTCAACAACGGATCTCTTGGTTCTGG CATCGATGAAGAACGCAGCAAAATGCGATAAGTAATGTGAATTGCAGAATTCAGTGAAT CATCGAATCTTTGAACGCACATTGCGCCCGCCAGTATTCTGGCGGGCATGCCTGTTCGAG CGTCATTTCAACCCTCAAGCACAGCTTGGTGTTGGGACTCGCGTTAATTCGCGTTCCCCA AATT...
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