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Fig. S1 Southern blotting analysis of the copy numbers of MoSWI6 in Magnaporthe oryzae. Genomic DNA of wild-type Guy11 was digested with SmaI, EcoRV and EcoRI, respectively, and separated in a 0.7% agarose gel. The DNA was hybridized with MoSWI6 gene probe amplified with primers FL3157/FL2911.

Fig. S2 Phylogenetic analysis of Magnaporthe oryzae Swi6 amino acid sequence with its homologues in other organisms. The neighbour-joining tree (with 1000 bootstrap replicates) of phylogenetic relationships between Swi6 homologues in fungi was constructed with the following results: M. oryzae (Magnaporthe oryzae XP_365024), G. zeae (Gibberella zeae XP_384396), P. anserina (Podospora anserina XP_001903283), N. crassa (Neurospora crassa XP_962967), C. globosum (Chaetomium globosum XP_001224444), B. fuckeliana (Botryotinia fuckeliana XP_001557910), S. sclerotiorum (Sclerotinia sclerotiorum XP_001590455), A. nidulans (Aspergillus nidulans XP_664319), A. niger (Aspergillus niger XP_001391313), A. fumigatus (Aspergillus fumigatus XP_748947), A. oryzae (Aspergillus oryzae XP_001817491), A. terreus (Aspergillus terreus XP_001215548), C. immitis (Coccidioides immitis XP_001246031), C. albicans (Candida albicans ACH78334) and S. cerevisiae (Saccharomyces cerevisiae NP_013283). Evolutionary distances are indicated by the scale bar below.

Fig. S3 Amino acid sequence alignment of APSES domain and ankyrin repeat (ANK repeat) of Magnaporthe oryzae with their homologues from other fungi. (A) The amino acid alignment of APSES domains from M. oryzae (Magnaporthe oryzae XP_365024), B. fuckeliana (Botryotinia fuckeliana XP_001557910), G. zeae (Gibberella zeae XP_384396), N. crassa (Neurospora crassa XP_962967), C. globosum (Chaetomium globosum XP_001224444), P. anserina (Podospora anserina XP_001903283), S. sclerotiorum (Sclerotinia sclerotiorum XP_001590455) and A. oryzae (Aspergillus oryzae XP_001817491). (B) Alignment of ANK repeats of MoSwi6 and its homologues from other fungi. Identical amino acids are highlighted with a black background and similar amino acids with a grey background.

Fig. S4 Targeted deletion of MoSWI6 in Magnaporthe oryzae. (A) Schematic illustration for MoSWI6 targeted gene replacement. The organization of the MoSWI6 locus and the gene deletion vector; the positions and orientations of the primers FL2790 (1), FL2791 (2), FL2792 (3) and FL2793 (4) are labelled with small arrows. The FL2790/FL2793 fragment amplified from the gene deletion vector was purified by gel electrophoresis and used to transform M. oryzae Guy11 protoplasts. (B) Mutant transformants were verified by Southern blot analysis. Genomic DNA was digested with EcoRI and separated on a 0.7% agarose gel. The DNA was hybridized with probe A, amplified with primers FL3157 and FL3197, and probe B, the 1.4-kb HPH fragment, amplified with primers FL1111 and FL1112. (C) Mutant transformants were verified by polymerase chain reaction (PCR). Transformants #3 and #5 were representative mutants. ΔMoswi6/MoSWI6 was obtained by transformation of #3 ΔMoswi6 strain with the wild-type MoSWI6 gene. pMD-Moswi6, plasmid of knockout construct; pMD-Moswi6/MoSWI6, plasmid of complemented construct. (D) Mutants were verified by reverse transcriptase-polymerase chain reaction (RT-PCR). The primers used as an endogenous control were specific to MoSWI6, using the same total RNA to the M. oryzae actin gene. PCR products were separated on an agarose gel and stained with ethidium bromide. Data represent three independent experiments, each performed three times and yielding similar results.

Fig. S5 Deletion MoSWI6 resulted in reduced growth on various media. The ΔMoswi6 mutants displayed retarded growth on CM, OMA, V8, MM and SDC media.

Fig. S6 The expression level of two melanin biosynthesis genes in the wild-type and ΔMoswi6 mutant. Asterisks indicate significant differences at P = 0.01.

Table S1 Effects of stress conditions on the growth-inhibiting rate of Guy11 and ΔMoswi6 mutantsa

Table S2 Polymerase chain reaction (PCR) primers used in this study.

Table S3 Quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) primers used in this study.

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