The study was to standardize days to root sampling of four tomato species under normal condition during kharif-2013 at University of Agriculture Sciences, Bangalore. The results indicated that there were significant differences among root and shoot traits sampled at 85 and 110 days after sowing (DAS). Moderate to high PCV and GCV were recorded for all root and shoot traits studied but higher magnitude of PCV as compared to GCV was found, especially for root volume. Highest heritability and moderate to high GAM was observed for root: shoot dry weight, followed by shoot: root length and root dry weight indicating suitable traits for selection. Four species also showed variation in morphological, yield related and fruit yield per plant. Using paired t-test to compare means of five roots and shoot traits between 60-85 and 85-110 DAS revealed that the root sampling at 85 DAS for S. lycopersicum and S. pimpinellifolium will be appropriate with expressing non-significance for all traits, viz., root length, root: shoot length ratio, root: shoot dry weight ratio and root volume, except root length of S. peruvianum between 85-110 DAS, while the remaining species, S. lycopersicum var. cerasiforme and S. peruvianum were at 110 DAS which exhibited significance for almost the traits, except root: shoot length ratio and root:shoot dry weight ratio of S. peruvianum and root: shoot length ratio of S. lycopersicum var. cerasiforme.