Characterization of representative transgenic tomato HMGS‐OEs. (a, b) Western blot analysis using antibodies against BjHMGS1 to verify the expression of BjHMGS1 in tomato leaves of 5‐week‐old representative wild‐type HMGS‐OEs (OE‐wtBjHMGS1) in (a) and mutant HMGS‐OEs (OE‐S359A) in (b). The cross‐reacting HMGS band is indicated by an arrowhead. Putative tomato HMGS‐OEs were designated as OE‐wtBjHMGS1 lines (413, 430, 439, 442, 444 and 445) in (a) and OE‐S359A lines (622, 623, 624, 625, 631 and 635) in (b). PC, positive control (tobacco BjHMGS1 OE line 402 as reported in Liao et al., 2014b); pSa13, vector (pSa13)‐transformed tomato. Bottom, Coomassie Blue‐stained gel of 20 μg total protein in each well. Two independent lines from each construct selected for further tests are underlined. White lines have been inserted between lanes that have been spliced together from the same original gel/blot. (c) Semiquantitative RT‐PCR analysis on representative transgenic tomato plants. BjHMGS1‐specific primers (ML1666 and ML1667) and tomato ACTIN (SlACTIN)‐specific primers (ML1688 and ML1689) were used. The PCR bands of 147‐bp BjHMGS1 and 176‐bp SlACTIN are indicated. pSa13, vector (pSa13)‐transformed control.