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Tomato cultivation is globally significant, demanding enhanced yields and biotic stress resilience for sustainability. Dwarf plant utilization in tomato genetic enhancement offers underexplored benefits. Yet, Santa Cruz dwarf tomato germplasm is unavailable. This study evaluated genetic dissimilarity, agronomic potential, and acylsugar content of Santa Cruz dwarf tomato plants across three successive backcrossings. Twelve advanced backcrossing populations (BC) and a commercial control (cv. Kada) were assessed, totaling 15 treatments. Agronomic traits and acylsugar content were measured, and analysis techniques were applied to assess genetic dissimilarity and backcrossing superiority. BC3 populations UFU-Sci#8, UFU-Sci#6, UFU-Sci#5, and UFU-Sci#1 excelled. Dwarf plants predominantly exhibited high leaflet acylsugar levels, suggesting potential for acquisition of hybrid with pest resistance. Notably, UFU MC TOM1 (dwarf plant) displayed significant glycine and L-serine presence, associated with various biotic stresses. These findings unveil the promising creation of dwarf Santa Cruz tomato hybrids with a broad spectrum of resistance. [ABSTRACT FROM AUTHOR]