Method of fungus inoculation
Potted plants of oilseed rape in the 5th leaf stage were used for the inoculation by the fungus. Only one leaf (4th leaf) per plant was inoculated by spraying it with the conidial suspension of the fungus strain (concentration: 5.0 × 107 conidia/ml). A small calibrated hand sprayer (100 ml capacity) was used for application of the required spray volume. The sprayer was standardized to apply fine jets of spray each of 0.5 ml spray volume. The upper and lower surfaces of the inoculated leaf were sprayed by applying two fine jets of spray per surface so that four sprays were applied per leaf with a total spray volume of 2.0 ml. During the inoculation process, care was taken to avoid the spray coming into contact with other plant parts by covering them carefully with aluminum foil. This precaution was undertaken to avoid contamination of other parts and to check whether the fungus moved and became endophytic in the unsprayed parts. For this purpose, re-isolation of the fungus from internal tissues distant from the initial site of inoculation (4th leaf) was attempted. Moreover, care was taken to ensure that the conidia did not splash from inoculated to non-inoculated leaves during the watering and fertilizing of plants which were accomplished by adding water directly to the soil surface and not spraying it onto the leaves. For the control plants of these bioassays, one leaf (4th leaf of each plant) was sprayed with a blank suspension (sterile distilled water + 0.1% Tween 20) instead of spraying the conidial suspension as was done for inoculated plants. The same spray volume as applied for inoculated plants was used. Moreover, considerable effort was exerted to avoid cross-contamination between plants (e.g. by covering the unsprayed portions of the plants; supplying water/fertilizer to the treated and control plants in a spatially segregated manner; fixing long, erect and thin transparent plastic sheets between potted plants to separate them from each other).
 
Method of fungus inoculationPotted plants of oilseed rape in the 5th leaf stage were used for the inoculation by the fungus. Only one leaf (4th leaf) per plant was inoculated by spraying it with the conidial suspension of the fungus strain (concentration: 5.0 × 107 conidia/ml). A small calibrated hand sprayer (100 ml capacity) was used for application of the required spray volume. The sprayer was standardized to apply fine jets of spray each of 0.5 ml spray volume. The upper and lower surfaces of the inoculated leaf were sprayed by applying two fine jets of spray per surface so that four sprays were applied per leaf with a total spray volume of 2.0 ml. During the inoculation process, care was taken to avoid the spray coming into contact with other plant parts by covering them carefully with aluminum foil. This precaution was undertaken to avoid contamination of other parts and to check whether the fungus moved and became endophytic in the unsprayed parts. For this purpose, re-isolation of the fungus from internal tissues distant from the initial site of inoculation (4th leaf) was attempted. Moreover, care was taken to ensure that the conidia did not splash from inoculated to non-inoculated leaves during the watering and fertilizing of plants which were accomplished by adding water directly to the soil surface and not spraying it onto the leaves. For the control plants of these bioassays, one leaf (4th leaf of each plant) was sprayed with a blank suspension (sterile distilled water + 0.1% Tween 20) instead of spraying the conidial suspension as was done for inoculated plants. The same spray volume as applied for inoculated plants was used. Moreover, considerable effort was exerted to avoid cross-contamination between plants (e.g. by covering the unsprayed portions of the plants; supplying water/fertilizer to the treated and control plants in a spatially segregated manner; fixing long, erect and thin transparent plastic sheets between potted plants to separate them from each other).
การแปล กรุณารอสักครู่..
