Method for direct transformation of exogenous DNA into resting spores of Penicillium amagasakiense

A method for direct transformation of exogenous DNA into resting spores of Penicillium amagasakiense, characterized in that, the method comprising steps of:

• 1) culture of Penicillium amagasakiense and collection of spores, wherein
• Penicillium amagasakiense is inoculated onto a surface of a solid agar medium, and cultured until Penicillium amagasakiense spores are overgrown on the surface of the medium, the Penicillium amagasakiense spores are washed off from the surface of the medium, suspension of the spores is aspirated off and filtered to remove mycelia, and filtrate containing the spores is collected, and centrifuged to collect the pelleted resting spores;
• 2) pretreatment of Penicillium amagasakiense spores, wherein
• the spores are re-suspended in an electroporation buffer, and centrifuged to collect the spore pellets, the re-suspension and centrifugation steps are repeated 3-4 times, and the last collected spore pellets are re-suspended in the electroporation buffer, to obtain an Penicillium amagasakiense spore suspension with a spore concentration of 104-1011 spores/ml,
• in which the electroporation buffer consists of 4-hydroxyethyl piperazineethanesulfonic acid having a final concentration of 0.01-100 mmol/L and mannitol having a final concentration of 0.5-5000 mmol/L, and the pH of the electroporation buffer is 3.0-9.5;
• and
• 3) electroporation of Penicillium amagasakiense spores by using HDEN method, wherein
• the Penicillium amagasakiense spore suspension prepared in the above steps and a plasmid to be transformed are added to wells of a cell culture plate and mixed uniformly, to obtain a mixture of the spores and the plasmid, the cell culture plate is placed on an ice bath for 10-15 min, subsequently electroporation is carried out by using the HDEN method using an Etta Biotech X-Porator H1 electroporator, by inserting an electroporator head fitted with a matrix electrode into the mixture of the spores and the plasmid, and energizing, to generate an electric field inside the mixture of the spores and the plasmid, the cell culture plate is placed on the ice bath again for 10-15 min after electroporation, and subsequently the mixture of the spores and the plasmid is aspirated off, to obtain resting spores of Penicillium amagasakiense with introduction of exogenous DNA,
• in which ratio of the Penicillium amagasakiense suspension to the plasmid to be transformed is 6-600000 μl of Penicillium amagasakiense spore suspension to 0.1-10000 μg of plasmid to be transformed;
• and
• wherein the parameters for the electroporation comprise:
• a voltage of 1-6000 V, pulse duration of 2-2000000 ms, and repeat for 1-100 times at an interval of 5-50000 ms.