Artificially domesticated phlcbopus portentosus strain HZ18006 and SSR marker fingerprint thereof
A technology of HZ18006, black boletus, applied in fungi, microorganism-based methods, and methods of using microorganisms, etc., can solve the problems of difficult to obtain strains, large individual differences, etc., to simplify harvesting procedures and labor, The effect of good image and stable genetic characteristics
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Embodiment 1
[0038] Obtain black boletus (Phlebopus portentosus) HZ18006 bacterial strain of the present invention after carrying out artificial domestication to wild black boletus fruiting body isolate bacterial strain:
[0039] 1. Specific methods of artificial domestication:
[0040] 1.1 Collection of wild fruiting bodies and strain acquisition: wild black cattle were collected in Jinghong, Mengla, Menghai of Xishuangbanna Dai Autonomous Prefecture, Yunnan Province, and Simao and Jinggu in Pu'er area (host trees include phoenix trees, pomelo trees and jackfruit trees) There were 100 fruiting bodies of Phlebopus portentosus, and the tissue pieces of the fruiting bodies were separated with PDA medium, and 100 wild strains were obtained after the mycelium was overgrown with the medium.
[0041] 1.2 Verification of fruiting ability: The 100 wild strains obtained are cultivated and fruited in a factory cultivation environment, and the strains with a fruiting rate of 50%-75% are selected for do...
Embodiment 2
[0051] The application of the strain HZ18006 in the industrial cultivation of edible fungi: The strain HZ18006 can easily complete the life cycle from the strain to the formation of basidiocarp on the artificial substrate, that is, the artificial cultivation of the fruiting body can be carried out through the strain under artificial conditions, and the Reach the purpose of harvesting black boletus fruiting bodies. Compared with other cultivated strains, it has excellent biological growth characteristics such as convenient fruiting body cultivation method, extensive management, fast fruiting, and high yield. The specific industrial cultivation application conditions are as follows:
[0052] 1. Make the most suitable medium M1 solid medium for HZ18006 strain: 20.0g potato, 20.0g glucose, 2.0g yeast extract, MgSO 4 1.0g, KH 2 PO 4 1.0g medium, 16.0g agar, 1000mL distilled water. M1 liquid medium does not add agar, and the other components are the same as M1 solid medium. On...
Embodiment 3
[0064] Embodiment 3: Construction of the SSR marker fingerprint of Boletus niger HZ18006 bacterial strain:
[0065] Through the development of microsatellite fingerprints, the genetic relationship analysis and species definition of HZ18006 strain and other black boletus strains can be accurately and quickly carried out:
[0066] 1. Extraction and detection of genomic DNA of Boletus nigricans HZ18006 strain: Omega mag-bind DNA kit was used to extract the total DNA of HZ18006 strain, and the operation method was carried out according to the instructions provided by the kit. The extracted DNA sample was mixed with 10X LoadingBuffer at a ratio of 10:1 and loaded on a 1% agarose gel for electrophoresis, and the quality of the extracted total DNA was detected using a gel imager. The concentration of total DNA was measured using a micro-volume ultraviolet spectrophotometer Nanodrop, and the concentration of sample DNA was adjusted to 50-100 ng / μL for PCR amplification reaction.
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