Soft rot caused by Pectobacterium carotovorum subsp. carotovorum is a destructive disease of the fleshy storage tissues of vegetables and ornamentals that occurs worldwide. This study was carried out to find an effective method for evaluating soft rot disease on a massive scale in greenhouse and to screen resistant accession for the disease with lettuce germplasm. Spray method with 5 mL suspension of 109 colony formation unit per milliliter (CFU/mL) of P. carotovorum subsp. carotovorum per plant at the 6 ~ 7 foliage leaf stage was the most effective to evaluate soft rot disease. We evaluated resistance to soft rot using this method for 812 accessions of lettuce germplasm introduced from 52 countries. As the results, withered rates for each germplasm accession ranged from 0 to 100 percent and among 812 accessions of lettuce germplasm investigated, 21 accessions showed very high resistance. Among the 21 accessions, 11 accessions were from Korea, each 2 accessions were from USA and Israel, and each 1 accession was from Afghanistan, Germany, Hungary, Netherlands, Pakistan, and Uzbekistan. Accessions from Israel, Syria, and Afghanistan showed high resistance level. Among very high resistant 21 accessions, 12 were landraces, 6 were cultivars, and 3 were wild relatives. The 21 accessions were consisted of 12 leaf lettuce, 5 romaine type, 3 wild relatives, and 1 butterhead lettuce. Leaf lettuce accessions showed the highest resistance level while crisphead lettuce accessions had the lowest resistance level among the cultivated lettuce types. Wild relatives had greater resistance than landraces and cultivars. These accessions which resistance to soft rot will be used for soft rot resistance breeding program in lettuce.