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. 2017 Oct 10;8:1953. doi: 10.3389/fmicb.2017.01953

FIGURE 1.

FIGURE 1

Complementation of ChMfs1 gene in C. higginsianum. (A) Strategic map of gene complementation vector construction and sites for restriction enzymes at the ChMfs1 genomic region. A bold line below the construct represents the sequence used as a probe (probe 1) in Southern blot analysis. (B) PCR analysis of wild-type Ch-1, T-DNA insertion mutant Ch-1-T513, and complementation strain C-ChMfs1-1. Marker in the image is DL15000. The 9406-bp band containing the sequences of ChMfs1 gene and inserted hph cassette from T-DNA insertion vector pTFCM was amplified in the mutant Ch-1-T513 and complementation strain C-ChMfs1-1 while a 3848-bp fragment containing ChMfs1 gene was obtained in the wild-type and C-ChMfs1-1. (C) Southern hybridization analysis of wild-type, T-DNA insertion mutant Ch-1-T513, and complementation strain C-ChMfs1-1. Genomic DNAs were digested with SacI and separated in 0.8% agarose gel. Blot was hybridized with the probe 1 amplified from genomic DNA of wild-type. (D) RT-PCR analysis of the transcription of wild-type Ch-1, T-DNA insertion mutant Ch-1-T513, and complementation strain C-ChMfs1-1. The 4-day old mycelia from PDA plates were collected and used for RNA isolation. Actin was used as the reference gene.