2.5. In vitro antifungal activity
The effects of Nitric oxide (0.1 mM SNP) on the in vitro growth of C. gloeosporioides and spore germination were tested following the method described by Zhang et al. (2013). Briefly, SNP solution or sterile water (control) was mixed with potato dextrose agar (PDA) to give a total volume of 15 mL per Petri plate (90 mm diameter). SNP concentrations in the PDA were 0 and 0.1 mM. After the PDA had solidified, an 8-mm diameter disk of mycelial mat from a 1-week-old culture was placed in the center of each Petri plate containing PDA and SNP. Mycelial growth as expressed by diameter (mm) was recorded after 2, 4, 6 and 8 d of incubation at 25 ± 1 ◦C. Each treatment concentration was replicated three times, and the experiment was repeated twice. For measurement of spore germination rate of C. gloeosporioides, aliquots of 100 L of spore suspension at 1 × 109 L−1 were individually transferred to glass tubes with potato dextrose broth (PDB), which contained SNP concentrations at 0 and 0.1 mM. All the tubes were put on a rotary shaker at 1.7 s−1 at 25 ◦C and incubated for up to 12 h. Approximately 200 spores were measured at 3 h intervals for germination rate per replicate, with 3 replicates for each treatment. The experiment was repeated twice.
2.5. In vitro antifungal activity The effects of Nitric oxide (0.1 mM SNP) on the in vitro growth of C. gloeosporioides and spore germination were tested following the method described by Zhang et al. (2013). Briefly, SNP solution or sterile water (control) was mixed with potato dextrose agar (PDA) to give a total volume of 15 mL per Petri plate (90 mm diameter). SNP concentrations in the PDA were 0 and 0.1 mM. After the PDA had solidified, an 8-mm diameter disk of mycelial mat from a 1-week-old culture was placed in the center of each Petri plate containing PDA and SNP. Mycelial growth as expressed by diameter (mm) was recorded after 2, 4, 6 and 8 d of incubation at 25 ± 1 ◦C. Each treatment concentration was replicated three times, and the experiment was repeated twice. For measurement of spore germination rate of C. gloeosporioides, aliquots of 100 L of spore suspension at 1 × 109 L−1 were individually transferred to glass tubes with potato dextrose broth (PDB), which contained SNP concentrations at 0 and 0.1 mM. All the tubes were put on a rotary shaker at 1.7 s−1 at 25 ◦C and incubated for up to 12 h. Approximately 200 spores were measured at 3 h intervals for germination rate per replicate, with 3 replicates for each treatment. The experiment was repeated twice.
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