The Background Akin To Fluorouracil

(2011). To harvest the plants, we carefully clipped the shoot and roots away from the rhizome. We also stained the roots of a subset of plants per treatment following Brundrett et al. (1984) to confirm RFS colonization. We then weighed the rhizome and immediately flash-froze it in liquid nitrogen. We stored samples at ?80 ��C until they could be lyophilized and ground. We followed the protocol of Zuleta and Sambucetti (2001) to analyse rhizome inulin (storage carbohydrate) and sucrose (mobile carbohydrate) content via high-performance liquid chromatography (HPLC). [Note: Starch is not present in the rhizome of Maianthemum (A. N. Hale et al., submitted Fluorouracil for publication).] In brief, a 0.03 g dried sample for each plant is boiled while stirring with a magnetic stir bar. Once samples cool to room temperature, they are filtered through a 0.20 ��m filter, and run on HPLC (Aminex HPX-87C anion-exchange column, deionized water at 85 ��C was set UNC2881 as the mobile phase with a flux rate of 0.6 mL min?1). Standards are used (inulin from dahlia tubers, Sigma-Aldrich; sucrose, Sigma-Aldrich) to confirm the identity of the sample peaks and to create standard curves to determine inulin and sucrose concentrations. Here, we express inulin and sucrose concentrations as a percentage of the HPLC dry sample mass. We also sum each plant's inulin and sucrose content to determine total NSC concentration (%). To explore the effect of our treatments on rhizome carbohydrate status, we use a multivariate analysis of covariance (MANCOVA). Following a significant MANCOVA, individual ANCOVA tests are conducted for inulin, sucrose and total NSC. For all models, we include harvest date as a main effect because rhizome carbohydrate concentration varies over the growing season in perennial herbs (e.g. Lapointe 1998; Wyka 1999; Kleijn et al. 2005). We also include initial plant mass as a covariate to account for differences in carbohydrate storage that are related to plant size/age (MANCOVA model: total NSC + inulin + sucrose = treatment + SRT1720 mw harvest date + initial plant mass; ANCOVA models: carbohydrate = treatment + harvest date + initial plant mass). We calculate least squares means and standard errors for all ANCOVA models with a significant (P