Plants from Phase 2 were evaluated for AMF colonisation using the acidified ink staining method of Vierheilig (1998). Plants were removed from their pot and roots washed free et al. of soil. A 2-5 g sub-sample from each root system was immersed in a 10% potassium hydroxide solution (10% w/v: 10g KOH in 100ml aqueous solution) and placed in a water bath at 80°C for 25 minutes. Thereafter roots were rinsed with water and blotted dry. Roots were placed in clean vials and covered with staining solution (84.4: 15: 0.6, dH2O: 1% HCI: Quink blue pen ink) in the water bath for a further 15 minutes. Colonisation was measured using the cross hair eye piece method of McGonigle (1990). et al. Statistical analysis All analyses were conducted using plants from Phase 2 as replicates. All data sets were checked for normality and homogeneity of variances prior to analyses and all percentage data were arc sine transformed. All data violating assumptions of homogeneity were transformed with logarithmic or reciprocal transformations. Linear mixed effect models (LMM) using the nlme package in R (Pinheiro 2012) was used to assess plant height and total leaf et al., number, as a response to the interaction between treatment and time. Plant replicate was assigned as a random effect. Exploratory analyses of the data revealed that the explanatory variable was quadratic. The quadratic term was only retained within the model ’time’ alongside the associated main effect when significant. A one-way ANOVA was used to examine soil treatment effects on fresh and dry biomass, as well as soil NO3, PO4 and K content and percentage root length colonized (% RLC) by AMF. One-way ANOVA’s were used to compare total mass of PLFAs, mass of bacteria attributed PLFAs alone and mass of fungal attributed PLFAs alone between invaded and native (control) soil. PLFA community analysis (Principal Component Analysis; PCA) was performed in R (R Core Team, 2014) by
Plants from Phase 2 were evaluated for AMF colonisation using the acidified ink staining method of Vierheilig et al. (1998). Plants were removed from their pot and roots washed free of soil. A 2-5 g sub-sample from each root system was immersed in a 10% potassium hydroxide solution (10% w/v: 10g KOH in 100ml aqueous solution) and placed in a water bath at 80°C for 25 minutes. Thereafter roots were rinsed with water and blotted dry. Roots were placed in clean vials and covered with staining solution (84.4: 15: 0.6, dH2O: 1% HCI: Quink blue pen ink) in the water bath for a further 15 minutes. Colonisation was measured using the cross hair eye piece method of McGonigle et al. (1990). Statistical analysis All analyses were conducted using plants from Phase 2 as replicates. All data sets were checked for normality and homogeneity of variances prior to analyses and all percentage data were arc sine transformed. All data violating assumptions of homogeneity were transformed with logarithmic or reciprocal transformations. Linear mixed effect models (LMM) using the nlme package in R (Pinheiro et al., 2012) was used to assess plant height and total leaf number, as a response to the interaction between treatment and time. Plant replicate was assigned as a random effect. Exploratory analyses of the data revealed that the explanatory variable ’time’ was quadratic. The quadratic term was only retained within the model alongside the associated main effect when significant. A one-way ANOVA was used to examine soil treatment effects on fresh and dry biomass, as well as soil NO3, PO4 and K content and percentage root length colonized (% RLC) by AMF. One-way ANOVA’s were used to compare total mass of PLFAs, mass of bacteria attributed PLFAs alone and mass of fungal attributed PLFAs alone between invaded and native (control) soil. PLFA community analysis (Principal Component Analysis; PCA) was performed in R (R Core Team, 2014) by
Pattison_et_al_2015_WR_PURE.docx