Evaluation of auxin-responsive genes in soybean for detection of off-target plant growth regulator herbicides

The expression of a candidate auxin-responsive gene was evaluated for use in a diagnostic assay for plant growth regulator (PGR) herbicide injury in soybean leaves. Expression of GH3, a primary auxin-responsive gene, was evaluated in response to dicamba and clopyralid at the RNA and protein levels, and proteomic analysis evaluated global expression of proteins in response to dicamba. Expression of GH3 was also analyzed in response to heat, drought, salt stress, and infection by soybean mosaic virus (SMV) and bean pod mottle virus (BPMV) to determine the specificity of GH3 expression as a diagnostic marker for PGR herbicide injury. At the RNA level, GH3 was strongly induced by dicamba and clopyralid within 8 h after application. Expression peaked 1 to 3 d after treatment (DAT) in response to 10% and 1% of a labeled dose of dicamba and clopyralid, with higher expression levels detected at higher herbicide rates. At the protein level, GH3 expression was also strongly induced at 1, 2, and 3 DAT by 10% vs. 1% of a labeled dose of dicamba and clopyralid. Heat, drought, and salt stress and infection with SMV or BPMV had no effect on GH3 expression at either the RNA or protein level. Proteomic analysis identified three proteins that were up-regulated in response to dicamba. Two were induced for less than 7 DAT and a third was identified as a stress-response enzyme (Superoxide dismutase) that is likely not specific to PGR herbicide injury. Expression of GH3 was highly induced by PGR herbicides at the RNA and protein level and was not affected by environmental stresses or viral infection, indicating that GH3 expression has excellent potential for use in a diagnostic assay for PGR herbicide injury.