Samples were resuspended in 20 ul of 80:20 MeOH:H2O per mg dry tissue and incubated at 4C for 10 min. Plant tissue was harvested, lyophilized to dryness, and homogenized using a ball mill (Retsch MM 400) at 25 Hz for 2 min. Forty-eight h later, the three treated lower leaves (local, L) and three untreated upper leaves (distal, D) (leaf number 8-10) were harvested, pooled, respectively, then frozen in liquid nitrogen for metabolic profiling by LC-MS, and triple quadrupole (QQQ)-MS analysis. Methods: Three lower leaves (leaf number 5-7) of 30-32-day-old Col-0, fmo1 and ugt76b1 Arabidopsis plants were infiltrated with 10 mM MgCl2 and a 5e6 cfu/ml suspension of Pst avrRpt2 in 10 mM MgCl2. Experimental Goal: To determine the metabolic profile of Arabidopsis WT, ugt76b1, and fmo1 leaves during systemic acquired resistance.
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