Trouble detecting infiltrated substrate in Nicotiana benthamiana (Agrobacterium system), works in vitro but not in planta Hi all, I’m running into an issue with substrate infiltration in *Nicotiana benthamiana* and would really appreciate any troubleshooting suggestions. **Setup:** * I transiently express my gene of interest via Agrobacterium infiltration. * After \~4 days of expression, I infiltrate an exogenous substrate into the leaves. * I then extract with ethyl acetate and analyze by GC-MS. **Problem:** * I cannot detect **either the infiltrated substrate or the expected product** in the extract. * This is surprising because: * The reaction works well in **crude protein extract (in vitro)**. * My extraction method seems fine, I can detect **products derived from endogenous** ***Nicotiana*** **substrates** using the same protocol. **Observations:** * The plants look **somewhat weak/stressed** after 4 days post-Agro infiltration.…