Abstract: In order to obtain the spoilage microorganisms in the swell barrelled tomato sauce,the main spoilage microorganisms in tomato sauce were separated and purified by gradient dilution method,selective culture method and scribing purification method. The morphological characteristics and 16S rDNA and ITS sequence analysis were used. The isolates were identified by combining physiological and biochemical identification as well. The results showed that the four bacteria numbered B001,B002,B003 and B004 included one Paenibacillus cineris,two Bacillus cereus,and one Bacillus mycoides. One yeast strain named F001 was Pichia kudriavzevil. Therefore,the main spoilage microorganisms in the swell barrelled tomato sauce should be Bacillus licheniformis,Bacillus cereus,Bacillus licheniformis and Pichia kudriavzevil.