Rusty J. Rodriguez Regina S. Redman 1994 <p><span>Factors affecting the efficient transformation of </span><i>Colletotrichum</i><span> species. </span><i>Experimental Mycology</i><span>, 18, 230-246. Twelve isolates representing four species of </span><i>Colletotrichum</i><span> were transformed either by enhanced protoplast, restriction enzyme-mediated integration (REMI), or electroporation-mediated protocols. The enhanced protoplast transformation protocol resulted in 100- and 50-fold increases in the transformation efficiencies of </span><i>Colletotrichum lindemuthianum</i><span> and </span><i>C. magna</i><span> , respectively. REMI transformation involved the use of </span><i>Hin</i><span> dIII and vector DNA linearized with </span><i>Hin</i><span>dIII to increase the number of integration events and potential gene disruptions in the fungal genome. Combining the enhanced protoplast and the REMI protocols resulted in a 22-fold increase in the number of hygromycin/nystatin-resistant mutants in </span><i>C. lindemuthianum</i><span> . Electroporation-mediated transformation was performed on mycelial fragments and spores of four </span><i>Colletotrichum</i><span> species, resulting in efficiencies of up to 1000 transformants/μg DNA. The pHA1.3 vector which confers hygromycin resistance contains telomeric sequences from </span><i>Fusarium oxysporum</i><span> , transforms by autonomous replication and genomic integration, and was essential for elevated transformation efficiencies of 100 to 10,000 transformants/μg DNA. Modifications of pHA1.3 occurred during bacterial amplification and post fungal transformation resulting in plasmids capable of significantly elevated transformation efficiencies in </span><i>C. lindemuthianum.</i></p> application/pdf 10.1006/emyc.1994.1023 en Elsevier Factors affecting the efficient transformation of Colletotrichum species article