Dissecting the Roles of Phosphorus Use Efficiency, Organic Acid Anions, and Aluminum-Responsive Genes under Aluminum Toxicity and Phosphorus Deficiency in Ryegrass Plants.

Aluminum (Al) toxicity and phosphorus (P) deficiency are widely recognized as major constraints to agricultural productivity in acidic soils. Under this scenario, the development of ryegrass plants with enhanced P use efficiency and Al resistance is a promising approach by which to maintain pasture production. In this study, we assessed the contribution of growth traits, P efficiency, organic acid anion (OA) exudation, and the expression of Al-responsive genes in improving tolerance to concurrent low-P and Al stress in ryegrass (Lolium perenne L.). Ryegrass plants were hydroponically grown under optimal (0.1 mM) or low-P (0.01 mM) conditions for 21 days, and further supplied with Al (0 and 0.2 mM) for 3 h, 24 h and 7 days. Accordingly, higher Al accumulation in the roots and lower Al translocation to the shoots were found in ryegrass exposed to both stresses. Aluminum toxicity and P limitation did not change the OA exudation pattern exhibited by roots. However, an improvement in the root growth traits and P accumulation was found, suggesting an enhancement in Al tolerance and P efficiency under combined Al and low-P stress. Al-responsive genes were highly upregulated by Al stress and P limitation, and also closely related to P utilization efficiency. Overall, our results provide evidence of the specific strategies used by ryegrass to co-adapt to multiple stresses in acid soils.

Biochemical and Molecular Responses Underlying the Contrasting Phosphorus Use Efficiency in Ryegrass Cultivars.

Improving plant ability to acquire and efficiently utilize phosphorus (P) is a promising approach for developing sustainable pasture production. This study aimed to identify ryegrass cultivars with contrasting P use efficiency, and to assess their associated biochemical and molecular responses. Nine ryegrass cultivars were hydroponically grown under optimal (0.1 mM) or P-deficient (0.01 mM) conditions, and P uptake, dry biomass, phosphorus acquisition efficiency (PAE) and phosphorus utilization efficiency (PUE) were evaluated. Accordingly, two cultivars with high PAE but low PUE (Ansa and Stellar), and two cultivars with low PAE and high PUE (24Seven and Extreme) were selected to analyze the activity and gene expression of acid phosphatases (APases), as well as the transcript levels of P transporters. Our results showed that ryegrass cultivars with high PAE were mainly influenced by root-related responses, including the expression of genes codifying for the P transporter LpPHT1;4, purple acid phosphatase LpPAP1 and APase activity. Moreover, the traits that contributed greatly to enhanced PUE were the expression of LpPHT1;1/4 and LpPHO1;2, and the APase activity in shoots. These outcomes could be useful to evaluate and develop cultivars with high P-use efficiency, thus contributing to improve the management of P in grassland systems.

Physiological and molecular insights involved in silicon uptake and transport in ryegrass.

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 Vmax values between cultivars; however, different Km suggested that Jumbo and Nui may have different affinities for silicic acid. The dissimilarities in Km 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.

Expression analysis and functional characterization of two PHT1 family phosphate transporters in ryegrass.

MAIN CONCLUSION: The phosphate transporters LpPHT1;1 and LpPHT1;4 have different roles in phosphate uptake and translocation in ryegrass under P stress condition. The phosphate transporter 1 (PHT1) family are integral membrane proteins that operate in phosphate uptake, distribution and remobilization within plants. In this study, we report on the functional characterization and expression of two PHT1 family members from ryegrass plants (Lolium perenne L.) and determine their roles in the specificity of Pi transport. The expression level of LpPHT1;4 was strongly influenced by phosphorus (P) status, being higher under P-starvation condition. In contrast, the expression level of LpPHT1;1 was not correlated with P supply. Yeast mutant complementation assays showed that LpPHT1;4 can complement the growth defect of the yeast mutant Δpho84 under Pi-deficient conditions, whereas the yeast mutant expressing LpPHT1;1 was not able to restore growth. Phylogenetic and molecular analyses indicated high sequence similarity to previously identified PHT1s from other species in the Poaceae. These results suggest that LpPHT1;1 may function as a low-affinity Pi transporter, whereas LpPHT1;4 could acts as a high-affinity Pi transporter to maintain Pi homeostasis under stress conditions in ryegrass plants. This study will form the basis for the long-term goal of improving the phosphate use efficiency of ryegrass plants.

Silicon-Mediated Alleviation of Aluminum Toxicity by Modulation of Al/Si Uptake and Antioxidant Performance in Ryegrass Plants.

Silicon (Si) has been well documented to alleviate aluminum (Al) toxicity in vascular plants. However, the mechanisms underlying these responses remain poorly understood. Here, we assessed the effect of Si on the modulation of Si/Al uptake and the antioxidant performance of ryegrass plants hydroponically cultivated with Al (0 and 0.2 mM) in combination with Si (0, 0.5, and 2.0 mM). Exposure to Al significantly increased Al concentration, mainly in the roots, with a consequent reduction in root growth. However, Si applied to the culture media steadily diminished the Al concentration in ryegrass, which was accompanied by an enhancement in root dry matter production. A reduced concentration of Si in plant tissues was also observed when plants were simultaneously supplied with Al and Si. Interestingly, Si transporter genes (Lsi1 and Lsi2) were down-regulated in roots after Si or Al was applied alone; however, both Lsi1 and Lsi2 were up-regulated as a consequence of Si application to Al-treated plants, denoting that there is an increase in Si requirement in order to cope with Al stress in ryegrass. Whereas Al addition triggered lipid peroxidation, Si contributed to an attenuation of Al-induced oxidative stress by increasing phenols concentration and modulating the activities of superoxide dismutase (SOD), catalase, peroxidase, and ascorbate peroxidase antioxidant enzymes. Differential changes in gene expression of SOD isoforms (Mn-SOD, Cu/Zn-SOD, and Fe-SOD) and the profile of peroxide (H2O2) generation were also induced by Si in Al-stressed plants. This, to the best of our knowledge, is the first study to present biochemical and molecular evidence supporting the effect of Si on the alleviation of Al toxicity in ryegrass plants.

Silicon in vascular plants: uptake, transport and its influence on mineral stress under acidic conditions.

MAIN CONCLUSION: So far, considerable advances have been achieved in understanding the mechanisms of Si uptake and transport in vascular plants. This review presents a comprehensive update about this issue, but also provides the new insights into the role of Si against mineral stresses that occur in acid soils. Such information could be helpful to understand both the differential Si uptake ability as well as the benefits of this mineral element on plants grown under acidic conditions. Silicon (Si) has been widely recognized as a beneficial element for many plant species, especially under stress conditions. In the last few years, great efforts have been made to elucidate the mechanisms involved in uptake and transport of Si by vascular plants and recently, different Si transporters have been identified. Several researches indicate that Si can alleviate various mineral stresses in plants growing under acidic conditions, including aluminium (Al) and manganese (Mn) toxicities as well as phosphorus (P) deficiency all of which are highly detrimental to crop production. This review presents recent findings concerning the influence of uptake and transport of Si on mineral stress under acidic conditions because a knowledge of this interaction provides the basis for understanding the role of Si in mitigating mineral stress in acid soils. Currently, only four Si transporters have been identified and there is little information concerning the response of Si transporters under stress conditions. More investigations are therefore needed to establish whether there is a relationship between Si transporters and the benefits of Si to plants subjected to mineral stress. Evidence presented suggests that Si supply and its subsequent accumulation in plant tissues could be exploited as a strategy to improve crop productivity on acid soils.