Effect of arsenate on endogenous levels of cytokinins with different existing forms in two Pteris species.

Our previous results showed that content of trans-zeatin (tZ) increases in leaves of heavy metal hyperaccumulators but decreases in non-hyperaccumulators growing in multiple heavy metal polluted soils. However, the relationship between arsenic (As) accumulation and endogenous forms of cytokinins (CTKs) in As hyperaccumulators remains unknown. Here a hydroponic experiment was conducted to compare the CTK forms in the As hyperaccumulator Pteris cretica var. nervosa and non-hyperaccumulator Pteris ensiformis under arsenate stress (0, 2, 5, and 10 mg L-1). A simple and cost-effective procedure for the determination of CTK forms in plants was established, and a stepwise regression analysis was used to study the relationship among total As contents and different forms of endogenous CTKs in fronds of two plants. The results showed that the optimized chromatographic parameters were Zobax SB-C18 column (5 μm × 4.6 mm × 250 mm), UV detection detector at 269 nm, a flow rate of 0.6 mL min-1, constant temperature of 45 °C and gradient elution with methanol-acetonitrile-1% acetic acid. Contents of chlorophylls in the fronds of P. ensiformis were significantly decreased with addition of As compared to P. cretica var. nervosa. Furthermore, the total As content in fronds of P. cretica var. nervosa was positively correlated to the contents of N6-(2-isopentenyl) adenine-7-β-D-glucoside (iP7G) and N6-(2-isopentenyl) adenosine (iPR). However, the total As content in fronds of P. ensiformis was negatively correlated to its trans-zeatin riboside (ZR) content. Therefore, iP7G and iPR could positively improve As accumulation by P. cretica var. nervosa.