AGROBACTERIUM-MEDIATED GENETIC TRANSFORMATION METHOD OF LEPTOSPHAERIA BIGLOBOSA

• Main Authors: HUANGFU, Haiyan, LI, Ziqin, SHI, Zhidan, JIA, Xiaoqing, ZHANG, Ying, ZHU, Chunxia, HUANGFU, Jiuru, YAN, Mengjiao, SONG, Peiling, HAO, Lifen, YANG, Yongqing, GUO, Chen, ZHAO, Lili
• Format: Patent
• Language: Chinese; English; French
• Published: 24.12.2020
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

Provided is an Agrobacterium-mediated genetic transformation method of Leptosphaeria biglobosa. The method comprises the steps of: 1) inoculating Agrobacterium containing a plasmid pCHs-GFP in an LB liquid medium for cultivation, and then transferring the same to a CM liquid medium containing 150-250 μg/mL of acetosyringone to resuspend the bacteria to obtain an Agrobacterium liquid; 2) collecting Leptosphaeria biglobosa conidia and mixing same with sterile water to prepare a conidia suspension with a concentration of (0.5-9)×106 cells/mL; and 3) mixing the Agrobacterium liquid with the conidia suspension, and after 3-4 days of co-cultivation, transferring the mixture to a screening medium to carry out screening cultivation for 7-15 days to obtain transformants. L'invention concerne un procédé de transformation génétique à médiation par Agrobacterium de Leptosphaeria biglobosa. Le procédé comprend les étapes suivantes : 1) inoculation d'Agrobacterium contenant un plasmide pCHs-GFP dans un milieu liquide LB po