Yellow flesh color in tomato is believed to be caused by abnormal transcript of phytoene synthase gene 1 (Psyl). However, our initial work indicated that a yellow fruited line PIll 4490 might have full length of genomic DNA sequence for Psyl. To understand the genetics of flesh color in PI114490, we crossed this line to red fruited lines, 0H9242 and LA3189, to develop F1 and F2 progenies, and sequenced the genomic DNA fragnlent of Psyl from Pll14490 and 0H9242. All F1 plants had red fruits, and F2 populations segregated as 3: 1 for red and yellow, indicating that one single recessive gene controlled the yellow flesh color in PI114490. This gene was allelic to the known r gene based on tests between PI114490 and four r mutants LA2056, LA3003, LA0353, and LA3532. Sequence analysis indicated that Pil14490 had a complete genomic DNA sequence for Psyl gene with only one nucleotide difference in the fourth intron: G in PI114490 and A in 0H9242. This substitution was associated with yellow flesh in the F2 population. There was no detectable or very low lycopene in PI114490 and all yellow fruited F2 individuals. Lycopene in the F1 (9.77 mg/100 g) was higher than the mean (6.52 mg/100g) of two parents, and ranged from O to 11.69 mg/100g in all red fruited F2 individuals. These results suggested that lycopene content was quantitatively inherited.