Table1 Growth and Sensitivities of the Mutant to Antifungal Reagent at Different Temperature

Supplements:

Fig. 1

CM SDS Congo red (μg﹒ml-1) Calcofluor white(μg﹒ml-1)

Fig. 1 Hyphal growth and sensitivity to antifungal reagents of the mutant at 25℃ and 35℃. Mycelial discs 5 mm in diameter were cut from 7-d-old fungal culture and placed at the center of a 90-mm Petri dishcontaining CM agar, with and without the antifungal reagents, cultured at 25℃(a) and 35℃(b) for 12 d.

Table1 Growth and sensitivities of the mutant to antifungal reagent at different temperature

Temperature
(℃) / stain / CM / SDS(%) / Congo red (μg﹒ml-1) / Calcofluor white (μg﹒ml-1)
0.005 / 0.01 / 0.02 / 0.04 / 50 / 100 / 200 / 50 / 100 / 200
25 / Wild-type / 0.61±0.05 / 0.56±0.05 / 0.51±0.04 / 0.46±0.04 / 0.26±0.02 / 0.57±0.05 / 0.59±0.05 / 0.52±0.04 / 0.55±0.05 / 0.52±0.04 / 0.46±0.04
man1-79 / 0.52±0.04 / 0.53±0.04 / 0.47±0.04 / 0.39±0.03 / 0.29±0.02 / 0.54±0.05 / 0.51±0.04 / 0.44±0.04 / 0.52±0.04 / 0.57±0.05 / 0.55±0.05
man1-39 / 0.56±0.05 / 0.58±0.05 / 0.44±0.04 / 0.46±0.04 / 0.28±0.02 / 0.51±0.04 / 0.50±0.04 / 0.56±0.05 / 0.51±0.04 / 0.58±0.05 / 0.47±0.04
35 / Wild-type / 0.20±0.02 / 0.18±0.01 / 0.16±0.01 / 0.15±0.01 / 0.13±0.02 / 0.18±0.02 / 0.17±0.01 / 0.18±0.01 / 0.18±0.01 / 0.18±0.02 / 0.18±0.01
man1-79 / 0.16±0.01 / 0.17±0.01 / 0.14±0.01 / 0.13±0.01 / 0.12±0.01 / 0.16±0.01 / 0.14±0.01 / 0.16±0.01 / 0.16±0.01 / 0.16±0.01 / 0.15±0.01
man1-39 / 0.18±0.01 / 0.15±0.01 / 0.14±0.01 / 0.13±0.01 / 0.12±0.01 / 0.18±0.01 / 0.16±0.01 / 0.16±0.01 / 0.19±0.02 / 0.18±0.01 / 0.16±0.01

Fig. 2

Fig. 2.Germination and septation of the mutant. Freshly harvested conidia (5×104 conidia ml-1) were prepared from 7-day-old rice polish agar cultures, and aliquots (15μl) were applied to the hydrophobic side of a GelBond film (Cambrex Bio-Science, Rockland, ME). The conidial droplets were incubated in apetri dish with wet filter paper at room temperature. The septum of conidial germination were stained with calcofluor white(10 mg·ml-1,Sigma) aftert 1,2,3,4 and 5h postincubation. The septum are marked with an arrow.

Fig. 3

Fig. 3. Hyphal growth of the mutant.The gene replacement mutant△man1-79, ectopic transformant man1-39 and wide type control Guy11 werecultivated on starch yeast medium at 25℃.Theglass coverslipwere inserted lightly into the medium.After incubation for 2 d,the glass coverslip covered with the mycelia were stained with calcofluor white(10 mg·ml-1,Sigma).The septum are marked with an arrow.