Quantifying Phytohormones and Sugars

Abscisic acid content was measured as described by Lim et al [51 (link)]. Crushed seed (0.1 g) was extracted in 2ml of methanol containing 500 mg/L citric acid monohydrate and 100 mg/L butylated hydroxyl toluene, at 4°C overnight on a rotary shaker. The mixture was centrifuged at 2000 × g for 10 min, and the supernatant was transferred and dried using a speed vac (CVE-2000, EYELA). Samples were quantified using a Phytodetek-ABA kit (Agdia Inc., USA) according to the manufacturer’s instructions. Gibberellic acid (GA, MBS9310617, MyBioSource) and sugars (K-SUFRG, Megazyme, Ireland), including sucrose, D-fructose, D-glucose, were evaluated according to the manufacturer’s protocol.