After spray applications, five leaves per treatment were
collected from greenhouse plants in the morning, rinsed and
disinfested as previously described (Francis et al., 2010). Leaves
were rinsed three times with sterile distilled water in the same
plastic bags to remove any debris or spray residues, dipped in 70%
ethanol for 30 s, immersed in 0.5% sodium hypochlorite for 30 s,
and then immediately rinsed three times with sterile distilled
water. Leaves handled by the petiole end placed on a sterile paper
towel and inoculated with 105 cfumL1 as described above. Excess
inoculum was wiped from the leaf surface with a sterile paper
towel. Inoculated leaves were placed on the surface of soft water
agar (0.5%) with the abaxial side up. The petiole was removed and
the leaf pressed onto the agar surface with a plastic spreader to
obtain as much contact as possible. Petri dishes were sealed with
Parafilm and incubated in an environmentally controlled growth
chamber under fluorescent light at 60 mmol m2 s1 for 12 h
photoperiod at 28 C. Symptoms on the inoculated detached
leaves were assessed 7, 10 and 14 days post inoculation (dpi). The
experiment was conducted three times with six replications per
treatment.
After spray applications, five leaves per treatment werecollected from greenhouse plants in the morning, rinsed anddisinfested as previously described (Francis et al., 2010). Leaveswere rinsed three times with sterile distilled water in the sameplastic bags to remove any debris or spray residues, dipped in 70%ethanol for 30 s, immersed in 0.5% sodium hypochlorite for 30 s,and then immediately rinsed three times with sterile distilledwater. Leaves handled by the petiole end placed on a sterile papertowel and inoculated with 105 cfumL1 as described above. Excessinoculum was wiped from the leaf surface with a sterile papertowel. Inoculated leaves were placed on the surface of soft wateragar (0.5%) with the abaxial side up. The petiole was removed andthe leaf pressed onto the agar surface with a plastic spreader toobtain as much contact as possible. Petri dishes were sealed withParafilm and incubated in an environmentally controlled growthchamber under fluorescent light at 60 mmol m2 s1 for 12 hphotoperiod at 28 C. Symptoms on the inoculated detachedleaves were assessed 7, 10 and 14 days post inoculation (dpi). Theexperiment was conducted three times with six replications pertreatment.
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