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Figure 1:
Effect of temperature and nutrients on rate of growth of Hyalorbilia strains DoUCR50, StM, and ARF in agar culture. Mycelial growth from blocks of PDA cultures placed on PDA or water agar and grown at 18, 23, 28, or 33°C was measured after 10 days. The experiment was repeated twice, and data were pooled. Standard error bars are shown.
Figure 2:
Maximum likelihood tree depicting the phylogenetic relationships of three nematophagous fungi and related members of the Orbiliomycetes. The outgroup is Tuber borchii. Bootstrap values (out of 100) are shown on the branches. Branch lengths are the expected number of substitutions per nucleotide.
Figure 3:
Growth of Hyalorbilia strains on 3-week-old Heterodera schachtii females. Three-week-old H. schachtii females were placed on water agar cultures of DoUCR50 (A), StM (B), or ARF (C). After 3 days, females were transferred to water agar and incubated for an additional 6 days at 23°C. Parasitized females were cut in half with a scalpel blade and incubated for 24 hr at 23°C prior to being photographed.
Figure 4:
Effect of Heterodera schachtii female age on susceptibility to J2 hatch suppression by Hyalorbilia strains DoUCR50, StM, and ARF. For each treatment, 25 H. schachtii females were placed on water agar (Control) or water agar cultures of fungi for 3 days at 23°C. Females were transferred to water agar plates, five females per plate, for an additional 6 days to monitor J2 hatch. The experiment was repeated twice, and the data were pooled. For each time point, treatments with the same letter are not significantly different (P < 0.05).
Figure 5:
Effect of Heterodera schachtii female age on susceptibility of isolated eggs to parasitism by DoUCR50, StM, or ARF. Five groups of 10 eggs from females collected 3, 4, or 5 weeks after inoculation of cabbage were spotted onto water agar cultures of the fungi or on water agar without fungus (Control). Plates were incubated for 10 days at 23°C in the dark, and the eggs were monitored for parasitism and hatching of J2. Data are reported as the average number of parasitized eggs per group of 10. The experiment was repeated twice, and the data were pooled. Standard error bars with the same lowercase letter indicate no significant difference among fungi of a respective week.
API ZYM assay for extracellular hydrolytic enzyme activity.
Treatmenta
Trypsin
Acid phosphatase
α-glucosidase
β-glucosidase
N-acetyl-β-glucosamidase
C PDA
0.0 ± 0.00 c
0.0 ± 0.00 b
0.0 ± 0.00 c
0.0 ± 0.00 c
0.0 ± 0.00 c
C Fem
0.0 ± 0.00 c
1.3 ± 0.33 ab
0.7 ± 0.67 bc
0.3 ± 0.33 bc
0.0 ± 0.00 c
DoUCR50 PDA
2.3 ± 0.33 b
3.7 ± 1.33 a
3.7 ± 0.33 a
1.7 ± 0.88 abc
1.3 ± 0.33 b
DoUCR50 Fem
3.7 ± 0.33 a
3.0 ± 0.00 ab
2.3 ± 0.67 ab
3.3 ± 0.33 a
3.3 ± 0.33 a
ARF PDA
0.0 ± 0.00 c
3.0 ± 0.00 ab
3.3 ± 0.33 a
1.3 ± 0.33 abc
1.7 ± 0.33 b
ARF Fem
0.0 ± 0.00 c
3.4 ± 0.33 ab
2.0 ± 0.58 abc
3.0 ± 0.00 ab
3.3 ± 0.33 a
StM PDA
0.0 ± 0.00 c
3.7 ± 1.33 a
3.3 ± 0.67 a
2.3 ± 0.33 ab
2.0 ± 0.58 ab
StM Fem
0.0 ± 0.00 c
3.4 ± 0.33 ab
1.0 ± 0.00 abc
2.7 ± 0.88 ab
3.3 ± 0.33 a
Percent nucleotide sequence identities of Hyalorbilia strains DoUCR50, StM, and ARF.
