Abstract

The silicon (Si) uptake system of two ryegrass (Lolium perenne L.) cultivars was characterised by assessing the concentration-and time-dependent kinetics. Additionally, a Si transporter gene was isolated from ryegrass and their expression pattern was analysed. The concentration-dependent kinetics was examined in Jumbo and Nui cultivars supplied with 0, 0.5, 1.0, 2.0, and 4.0 mM Si and harvested at 24 h and 21 d. The time-dependent kinetics was evaluated at 0, 0.5, or 2 mM Si doses after 0, 3, 6, 9, 12, and 24 h. RACE-PCR was performed to isolate a full-length sequence codifying for a Si transporter, and semi-quantitative and quantitative RT-PCR was used to analyse its expression pattern. Differential Si uptake between ryegrass cultivars was found. Moreover, Lineweaver-Burk linearization showed similar V-max values between cultivars; however, different K-m suggested that Jumbo and Nui may have different affinities for silicic acid. The dissimilarities in K-m between cultivars might involve either the differential contribution of known proteins responsible for Si uptake and transport or the involvement of undiscovered Si transporters. We identified a putative Si transporter from ryegrass Nui (LpLsi1), which was only expressed in roots and down-regulated by Si supply. The predicted amino acid sequence of LpLsi1 did not only show a high similarity and close phylogenetic relationship with monocot Si influx transporters but also indicated that it is a membrane protein possessing a high conservation of domains essential for silicic acid selectivity. Our findings provide evidence of LpLsi1 in ryegrass, which supports its high Si accumulation capacity.