Abstract：Black spot is one of the most severe diseases affecting cucumber（Cucumis sativus L.）production．It is an essential step to set up a methodology for artificial seedling identification of cucumber resistance to black spot in disease resistance breeding．Pycnoconidia spore suspension with concentration 1×107-2×107 per mL was used as identification method of resistance to cucumber black spot．The spore suspension was sprayed on to the seedlings with 1 or 2 true leaf，and incubated at 22 ℃ in growth chamber under humid conditions for 48 hours and keeping relative humidity 100%，then kept in the condition（day 28-30 ℃，night 20 ℃，13 hours light per day，11 hours in dark for 5-15 days）before being evaluated by scoring the disease-lesion on each plant leaf．The inoculation results showed that 10 accessions were high resistant and 8 accessions were mid-resistant to black spot．D9 and W3 were high resistant varieties and 3-1，LW11，BA74，A8674 were resistant varieties to black spot．
