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emi2871-sup-0001-si.doc713K

Table S1. The primers used for the manipulation of Ras1 and Ras2 genes in B. bassiana.

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Table S2. Primers used to assess the transcript levels of downstream effector genes in the Ras1 and Ras2 mutants of B. bassiana via qRT-PCR.

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Fig. S1. Construction and identification of B. bassiana Ras1 and Ras2 mutants.

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A. Diagram for Ras2 knockout (see Table S1 for the used primers).

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B. PCR detection for the disrupted and complemented Ras2 in the genomic DNAs of wild type (lanes 1 and 4), ΔRas2 (lanes 2 and 5) and ΔRas2/Ras2 (lanes 3 and 6) with paired primers R2id-F/R (lanes 1−3) and Sur2-F/R (lanes 4−6).

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C. Southern blotting of Ras2 in the XbaI-digested DNA extracts from wild type (lane 1), ΔRas2 (lanes 2) and ΔRas2/Ras2 (lanes 3) using the probe amplified with R2S-F/R.

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D and E. PCR detection for the presence of Ras1 (with partial PgpdA) and sur in the 3-day SDAY colonies of wild type (lane 1), mRas1G19V (lanes 2−4), mRas1D126A (lanes 5−7), mRas1 (lanes 8−10), ΔRas2/Ras1G19V (lanes 11−13), ΔRas2/Ras1D126A (lanes 14−16) and ΔRas2/Ras1 (lanes 17−19) respectively.

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