Silicon Transporter Research Articles

Molecular forecasting of domoic acid during a pervasive toxic diatom bloom

In 2015, the largest recorded harmful algal bloom (HAB) occurred in the Northeast Pacific, causing nearly 100 million dollars in damages to fisheries and killing many protected marine mammals. Dominated by the toxic diatom Pseudo-nitzschia australis, this bloom produced high levels of the neurotoxin domoic acid (DA). Through molecular and transcriptional characterization of 52 near-weekly phytoplankton net-tow samples collected at a bloom hotspot in Monterey Bay, California, we identified active transcription of known DA biosynthesis (dab) genes from the three identified toxigenic species, including P. australis as the primary origin of toxicity. Elevated expression of silicon transporters (sit1) during the bloom supports the previously hypothesized role of dissolved silica (Si) exhaustion in contributing to bloom physiology and toxicity. We find that coexpression of the dabA and sit1 genes serves as a robust predictor of DA one week in advance, potentially enabling the forecasting of DA-producing HABs. We additionally present evidence that low levels of iron could have colimited the diatom population along with low Si. Iron limitation represents an overlooked driver of both toxin production and ecological success of the low-iron-adapted Pseudo-nitzschia genus during the 2015 bloom, and increasing pervasiveness of iron limitation may fuel the escalating magnitude and frequency of toxic Pseudo-nitzschia blooms globally. Our results advance understanding of bloom physiology underlying toxin production, bloom prediction, and the impact of global change on toxic blooms.

Characterization and function of promoters of silicon transporter genes PeLsi1-1 and PeLsi1-2 from moso bamboo (Phyllostachys edulis).

Promoters of moso bamboo silicon transporter genes PeLsi1-1 and PeLsi1-2 contain elements in response to hormone, silicon, and abiotic stresses, and can drive the expression of PeLsi1-1 and PeLsi1-2 in transgene Arabidopsis. Low silicon 1 (Lsi1) transporters from different species have been shown to play an important role in influxing silicon from soil. In previous study, we cloned PeLsi1-1 and PeLsi1-2 from Phyllostachys edulis and verified that PeLsi1-1 and PeLsi1-2 have silicon uptake ability. Furthermore, in this study, the promoters of PeLsi1-1(1910bp) and PeLsi1-2(1922bp) were cloned. Deletion analysis identified the key regions of the PeLsi1-1 and PeLsi1-2 promoters in response to hormone, silicon, and abiotic stresses. RT-qPCR analysis indicated that PeLsi1-1 and PeLsi1-2 were regulated by hormones, salt stress and osmotic stress. In addition, we found that the driving activity of the PeLsi1-1 and PeLsi1-2 promoters was regulated by 2mM K2SiO3 and PeLsi1-1-P3 ~ P4 and PeLsi1-2-P4 ~ 5 were the regions regulated by silicon. Overexpression of PeLsi1-1 or PeLsi1-2 driven by 35S promoter in Arabidopsis resulted in a threefold increase of Si accumulation, whereas transgenic plants showed deleterious symptoms and dwarf seedlings and shorter roots under 2mM Si treatment. When the 35S promoter was replaced by PeLsi1-1 or PeLsi1-2 promoter, a similar Si absorption was achieved and the transgene plants grew normally. This study, therefore, demonstrates that the promoters of PeLsi1-1 and PeLsi1-2 are indeed effective in driving the expression of moso bamboo Lsi1 genes and leading to silicon uptake.

The allelopathic effects of Heterosigma akashiwo on Skeletonema costatum: Insights from gene expression and metabolomics analysis

The globally distributed harmful algal blooms (HAB) species, Heterosigma akashiwo, has been found to exhibit ichthyotoxicity. Previous studies have shown that H. akashiwo achieves a competitive edge during bloom occurrences by inhibiting the growth of a coexisting diatom, Skeletonema costatum, through allelopathy. However, the specific allelopathic mechanisms underlying the allelopathic effects of H. akashiwo on S. costatum remain unknown. To bridge this gap, our study utilized a combination of quantitative real-time PCR and metabolomics to examine the allelopathic processes of H. akashiwo on S. costatum. Our results demonstrate that the growth of S. costatum is hindered when co-cultured with H. akashiwo (initial cell concentration, 2 × 104 cell/mL). Gene expression investigation showed a substantial reduction in the mRNA levels of cytochrome b6, ribulose bisphosphate carboxylase large chain, and silicon transporter in S. costatum when grown in co-culture conditions. Furthermore, metabolic pathway analysis suggested that the allelopathic effects of H. akashiwo disrupted several vital metabolic pathways in S. costatum, including a reduction in purine and pyrimidine metabolism and an increase in fatty acid biosynthesis. Our investigation has revealed the intricate and substantial involvement of allelopathy in the formation of H. akashiwo blooms, demonstrating the complexity of the allelopathic interaction between H. akashiwo and S. costatum. These insights also contribute significantly to our understanding of the dynamics within HAB species.

The draft genome of Nitzschia closterium f. minutissima and transcriptome analysis reveals novel insights into diatom biosilicification

BackgroundNitzschia closterium f. minutissima is a commonly available diatom that plays important roles in marine aquaculture. It was originally classified as Nitzschia (Bacillariaceae, Bacillariophyta) but is currently regarded as a heterotypic synonym of Phaeodactylum tricornutum. The aim of this study was to obtain the draft genome of the marine microalga N. closterium f. minutissima to understand its phylogenetic placement and evolutionary specialization. Given that the ornate hierarchical silicified cell walls (frustules) of diatoms have immense applications in nanotechnology for biomedical fields, biosensors and optoelectric devices, transcriptomic data were generated by using reference genome-based read mapping to identify significantly differentially expressed genes and elucidate the molecular processes involved in diatom biosilicification.ResultsIn this study, we generated 13.81 Gb of pass reads from the PromethION sequencer. The draft genome of N. closterium f. minutissima has a total length of 29.28 Mb, and contains 28 contigs with an N50 value of 1.23 Mb. The GC content was 48.55%, and approximately 18.36% of the genome assembly contained repeat sequences. Gene annotation revealed 9,132 protein-coding genes. The results of comparative genomic analysis showed that N. closterium f. minutissima was clustered as a sister lineage of Phaeodactylum tricornutum and the divergence time between them was estimated to be approximately 17.2 million years ago (Mya). CAFF analysis demonstrated that 220 gene families that significantly changed were unique to N. closterium f. minutissima and that 154 were specific to P. tricornutum, moreover, only 26 gene families overlapped between these two species. A total of 818 DEGs in response to silicon were identified in N. closterium f. minutissima through RNA sequencing, these genes are involved in various molecular processes such as transcription regulator activity. Several genes encoding proteins, including silicon transporters, heat shock factors, methyltransferases, ankyrin repeat domains, cGMP-mediated signaling pathways-related proteins, cytoskeleton-associated proteins, polyamines, glycoproteins and saturated fatty acids may contribute to the formation of frustules in N. closterium f. minutissima.ConclusionsHere, we described a draft genome of N. closterium f. minutissima and compared it with those of eight other diatoms, which provided new insight into its evolutionary features. Transcriptome analysis to identify DEGs in response to silicon will help to elucidate the underlying molecular mechanism of diatom biosilicification in N. closterium f. minutissima.

Synergy of plant growth promoting rhizobacteria and silicon in regulation of AgNPs induced stress of rice seedlings

Silver Nanoparticles (AgNPs), as an emerging pollutant, have been receiving significant attention as they deepen the concern regarding the issue of food security. Silicon (Si) and plant growth-promoting rhizobacteria (PGPR) are likely to serve as a sustainable approach to ameliorating abiotic stress and improving plant growth through various mechanisms. The present study aims to evaluate the synergistic effect of Si and PGPRs on growth, physiological, and molecular response in rice seedlings (Oryza sativa) under AgNPs stress. Data suggested that under AgNPs exposure, the root and shoot growth, photosynthetic pigments, antioxidant enzymes (CAT and APX), expression of antioxidant genes (OsAPX and OsGR), silicon transporter (OsLsi2), and auxin hormone-related genes (OsPIN10 and OsYUCCA1) were significantly decreased which accompanied with the overproduction of reactive oxygen species (ROS), nitric oxide (NO) and might be due to higher accumulation of Ag in plant cells. Interestingly, the addition of Si along with the AgNPs enhances the level of ROS generation, thus oxidative stress, which causes severe damage in all the above-tested parameters. On the other hand, application of PGPR alone and along with Si reduced the toxic effect of AgNPs through the improvement of growth, biochemical, and gene regulation (OsAPX and OsGR, OsPIN10 and OsYUCCA1). However, the addition of L-NAME along with PGPR and silicon drastically lowered the AgNPs induced toxicity through lowering the oxidative stress and maintained the overall growth of rice seedlings, which suggests the role of endogenous NO in Si and PGPRs mediated management of AgNPs toxicity in rice seedlings.

Silicon transporters in plants: Unravelling the molecular Nexus with sodium and potassium transporters under salinity stress

Salt stress poses a serious hazard to plant growth by altering osmotic and ionic homeostasis, producing too many oxidants and radicals, and harming vital metabolic processes like photosynthesis. Plants use mechanistic cascades of biochemical and physiological processes to battle salt stress and prevent ion toxicity; nevertheless, repeated exposure can overwhelm the defence system, leading to plant death. The Salt-Overly Sensitive (SOS) pathway, which predominantly relies on Na+ exclusion from the cytosol, makes a significant contribution to salinity tolerance in plants. Although silicon (Si) is known to reduce salt stress in a variety of crops and to raise plant stress tolerance, its impact on Na+ transport is little understood. In this review, we emphasise recent research on the interaction between Si treatment and important Na+ and K+ transporters involved in ion homeostasis under salt stress. The following aspects will receive special consideration: (1) The effects of salinity on membrane stability and ion homeostasis and the involvement of Na+ and K+ transporters in ion homeostasis (2) The uptake, storage, and transport of Si in higher plants, as well as the discovered Si transporters in many plant species (3) Modulation of the expression of the Na+, K+, and Si transporters to affect the absorption, transport, and homeostasis of ions by Si. Finally, this review also highlights the necessity for further investigation into the function of Si in salt stress in plants and the discovery of knowledge gaps in the broader area of this process.

Editing Silicon Transporter Genes to Reduce Arsenic Accumulation in Rice.

Rice is a dominant source of inorganic arsenic (As) exposure for populations consuming rice as a staple food. Decreasing As accumulation in rice grain is important for improving food safety. Arsenite [As(III)], the main form of As in paddy soil porewater, is taken up inadvertently by OsLsi1 and OsLsi2, the two key transporters for silicon (Si) uptake in rice roots. Here, we investigated whether editing OsLsi1 or OsLsi2 can decrease As accumulation in rice grain without compromising grain yield. We used the CRISPR-Cas9 technology to edit the promoter region of OsLsi1 and the C-terminal coding sequence of OsLsi1 and OsLsi2, and we generated a total of 27 mutants. Uptake and accumulation of Si and As were evaluated in both short-term hydroponic experiments and in a paddy field. Deletion of 1.2-2 kb of the OsLsi1 promoter suppressed OsLsi1 expression in roots and Si uptake markedly and did not affect As(III) uptake or grain As concentration. Some of the OsLsi1 and OsLsi2 coding sequence mutants showed large decreases in the uptake of Si and As(III) as well as large decreases in Si accumulation in rice husks. However, only OsLsi2 mutants showed significant decreases (by up to 63%) in the grain total As concentration. Editing OsLsi2 mainly affected the accumulation of inorganic As in rice grain with little effect on the accumulation of dimethylarsenate (DMA). Grain yields of the OsLsi2 mutants were comparable to those of the wild type. Editing OsLsi2 provides a promising way to reduce As accumulation in rice grain without compromising the grain yield.

Gene Duplication, Shifting Selection, and Dosage Balance of Silicon Transporter Proteins in Marine and Freshwater Diatoms.

Numerous factors shape the evolution of protein-coding genes, including shifts in the strength or type of selection following gene duplications or changes in the environment. Diatoms and other silicifying organisms use a family of silicon transporters (SITs) to import dissolved silicon from the environment. Freshwaters contain higher silicon levels than oceans, and marine diatoms have more efficient uptake kinetics and less silicon in their cell walls, making them better competitors for a scarce resource. We compiled SITs from 37 diatom genomes to characterize shifts in selection following gene duplications and marine-freshwater transitions. A deep gene duplication, which coincided with a whole-genome duplication, gave rise to two gene lineages. One of them (SIT1-2) is present in multiple copies in most species and is known to actively import silicon. These SITs have evolved under strong purifying selection that was relaxed in freshwater taxa. Episodic diversifying selection was detected but not associated with gene duplications or habitat shifts. In contrast, genes in the second SIT lineage (SIT3) were present in just half the species, the result of multiple losses. Despite conservation of SIT3 in some lineages for the past 90-100 million years, repeated losses, relaxed selection, and low expression highlighted the dispensability of SIT3, consistent with a model of deterioration and eventual loss due to relaxed selection on SIT3 expression. The extensive but relatively balanced history of duplications and losses, together with paralog-specific expression patterns, suggest diatoms continuously balance gene dosage and expression dynamics to optimize silicon transport across major environmental gradients.

Genome-wide identification and characterisation of Aquaporins in Rosa chinensis

ABSTRACT Aquaporins (AQPs) are small integral membrane proteins that facilitate the efficient transport of small solutes like water, metalloids, urea, and CO2 across the membrane. Plant aquaporins are known to be involved in vital physiological processes like seed germination, stomatal movement, cell elongation, and reproductive growth. In the present study, 36 AQPs were identified through genome-wide computational analysis performed in the rose genome. Subsequent phylogenetic analysis classified the AQPs into five subfamilies, including 10 Plasma membrane intrinsic proteins (PIPs), 8 Tonoplast intrinsic proteins (TIPs), 9 Nodulin intrinsic proteins (NIPs), 3 Small intrinsic proteins (SIPs), and 6 uncharacterised intrinsic proteins (XIPs). Based on the prediction of silicon solute specificity, one of the NIPs is predicted to be a silicon transporter. Structural analysis revealed the characteristic six trans-membrane domains and two-half alpha helix harbouring two conserved NPA motifs in the majority of the predicted AQPs. The protein tertiary structures predicted with homology-based modelling were used to study the pore lining remains and to predict the solute transport activity of the AQPs. Expression profiling of the rose AQPs showed tissue-specific expression of many members. The data provided here will help in understanding the AQP-based solute transport system in roses and other related species.

A silicon transporter gene required for healthy growth of rice on land

Silicon (Si) is the most abundant mineral element in the earth’s crust. Some plants actively accumulate Si as amorphous silica (phytoliths), which can protect plants from stresses. Here, we report a gene (SIET4) that is required for the proper accumulation and cell-specific deposition of Si in rice and show that it is essential for normal growth. SIET4 is constitutively expressed in leaves and encodes a Si transporter. SlET4 polarly localizes at the distal side of epidermal cells and cells surrounding the bulliform cells (motor cells) of the leaf blade, where Si is deposited. Knockout of SIET4 leads to the death of rice in the presence but not absence of Si. Further analysis shows that SIET4 knockout induces abnormal Si deposition in mesophyll cells and the induction of hundreds of genes related to various stress responses. These results indicate that SIET4 is required for the proper export of Si from leaf cells to the leaf surface and for the healthy growth of rice on land.

Molecular Characterization of Silicon (Si) Transporter Genes, Insights into Si-acquisition Status, Plant Growth, Development, and Yield in Alfalfa

Molecular Characterization of Silicon (Si) Transporter Genes, Insights into Si-acquisition Status, Plant Growth, Development, and Yield in Alfalfa

A citrate efflux transporter important for manganese distribution and phosphorus uptake in rice.

The plant citrate transporters, functional in mineral nutrient uptake and homeostasis, usually belong to the multidrug and toxic compound extrusion transporter family. We identified and functionally characterized a rice (Oryza sativa) citrate transporter, OsCT1, which differs from known plant citrate transporters and is structurally close to rice silicon transporters. Domain analysis depicted that OsCT1 carries a bacterial citrate-metal transporter domain, CitMHS. OsCT1 showed citrate efflux activity when expressed in Xenopus laevis oocytes and is localized to the cell plasma membrane. It is highly expressed in the shoot and reproductive tissues of rice, and its promoter activity was visible in cells surrounding the vasculature. The OsCT1 knockout (KO) lines showed a reduced citrate content in the shoots and the root exudates, whereas overexpression (OE) line showed higher citrate exudation from their roots. Further, the KO and OE lines showed variations in the manganese (Mn) distribution leading to changes in their agronomical traits. Under deficient conditions (Mn-sufficient conditions followed by 8 days of 0 μm MnCl2 · 4H2 O treatment), the supply of manganese towards the newer leaf was found to be obstructed in the KO line. There were no significant differences in phosphorus (P) distribution; however, P uptake was reduced in the KO and increased in OE lines at the vegetative stage. Further, experiments in Xenopus oocytes revealed that OsCT1 could efflux citrate with Mn. In this way, we provide insights into a mechanism of citrate-metal transport in plants and its role in mineral homeostasis, which remains conserved with their bacterial counterparts.

Physiological and Molecular Analysis Revealed the Role of Silicon in Modulating Salinity Stress in Mung Bean

Salinity stress acts as a significant deterrent in the course of optimal plant growth and productivity, and mung bean, being a relay crop in the cereal cropping system, is severely affected by salinity. Silicon (Si), on the other hand, has exhibited promising outcomes with regards to alleviating salinity stress. In order to understand the critical mechanisms underlying mung bean (Vigna radiata L.) tolerance towards salt stress, this study examined the effects of different salinity concentrations on antioxidant capacity, proteome level alterations, and influence on Si-transporter and salt-responsive genes. Salinity stress was seen to effect the gaseous exchange machinery, decrease the soluble protein and phenolic content and NR activity, and increase the accumulation of reactive oxygen species. An efficient regulation of stomatal opening upon Si application hints towards proficient stomatal conductance and CO2 fixation, resulting in efficient photosynthesis leading to proficient plant growth. The soluble protein and phenolic content showed improved levels upon Si supplementation, which indicates an optimal solute transport system from source to sink. The content of superoxide radicals showed a surge under salinity stress treatment, but efficient scavenging of superoxide radicles was noted under Si supplementation. Salinity stress exhibited more damaging effects on root NR activity, which was notably enhanced upon Si supplementation. Moreover, the beneficial role of Si was further substantiated as there was notable Si accumulation in the leaves and roots of salinity-stressed mung bean plants. Furthermore, Si stimulated competent ROS scavenging by reinforcing the antioxidant enzyme activity, as well coordinating with their isozyme activity, as expressed by the varying band intensities. Similarly, the Si-mediated increase in peroxidase activity may reveal changes in the mechanical characteristics of the cell wall, which are in turn associated with salinity stress adaptation. Proteomic investigations revealed the upregulation or downregulation of several proteins, which were thereafter identified by LC−MS/MS. About 45 proteins were identified and were functionally classified into photosynthesis (24%), metabolic process (19%), redox homeostasis (12%), transmembrane transport (10%), stress response (7%), and transcription regulation (4%). The gene expression analysis of the silicon transporter genes (Lsi1, Lsi2, and Lsi3) and SOS pathway genes (SOS1, SOS2, and SOS3) indicated the role of silicon in mitigating salinity stress. Hence, the findings of this study can facilitate a profound understanding of the potential mechanisms adopted by mung bean due to exogenous Si application during salinity stress.

Silicate solubilizing and plant growth promoting bacteria interact with biogenic silica to impart heat stress tolerance in rice by modulating physiology and gene expression.

Heat stress caused due to increasing warming climate has become a severe threat to global food production including rice. Silicon plays a major role in improving growth and productivity of rice by aiding in alleviating heat stress in rice. Soil silicon is only sparingly available to the crops can be made available by silicate solubilizing and plant-growth-promoting bacteria that possess the capacity to solubilize insoluble silicates can increase the availability of soluble silicates in the soil. In addition, plant growth promoting bacteria are known to enhance the tolerance to abiotic stresses of plants, by affecting the biochemical and physiological characteristics of plants. The present study is intended to understand the role of beneficial bacteria viz. Rhizobium sp. IIRR N1 a silicate solublizer and Gluconacetobacter diazotrophicus, a plant growth promoting bacteria and their interaction with insoluble silicate sources on morpho-physiological and molecular attributes of rice (Oryza sativa L.) seedlings after exposure to heat stress in a controlled hydroponic system. Joint inoculation of silicates and both the bacteria increased silicon content in rice tissue, root and shoot biomass, significantly increased the antioxidant enzyme activities (viz. superoxidase dismutase, catalase and ascorbate peroxidase) compared to other treatments with sole application of either silicon or bacteria. The physiological traits (viz. chlorophyll content, relative water content) were also found to be significantly enhanced in presence of silicates and both the bacteria after exposure to heat stress conditions. Expression profiling of shoot and root tissues of rice seedlings revealed that seedlings grown in the presence of silicates and both the bacteria exhibited higher expression of heat shock proteins (HSPs viz., OsHsp90, OsHsp100 and 60 kDa chaperonin), hormone-related genes (OsIAA6) and silicon transporters (OsLsi1 and OsLsi2) as compared to seedlings treated with either silicates or with the bacteria alone. The results thus reveal the interactive effect of combined application of silicates along with bacteria Rhizobium sp. IIRR N1, G. diazotrophicus inoculation not only led to augmented silicon uptake by rice seedlings but also influenced the plant biomass and elicited higher expression of HSPs, hormone-related and silicon transporter genes leading to improved tolerance of seedling to heat stress.

Strategies for Reducing Arsenic Content in Rice: A review

Arsenic (As) is one of the most toxic metalloid that can enter the food chain through ingestion of As contaminated water or food, posing a serious threat to human health. Among cereals, rice could contain the highest amount of As because of the special growing conditions. Therefore, the importance of the reduction of As concentration in rice is essential. Many studies have been conducted to understand the mechanism of arsenic uptake, accumulation and translocation. The interactions between As and plants are influenced by soil type and other factors such as pH, mineral contents and redox status of the soil, As speciation, and microbial activity. Different nutrients including phosphates, iron, silicon and sulfur effectively regulate the uptake and accumulation of As in different parts of plants. Genetic variation has also effect on As accumulation of rice grain. Water management practices can help to decrease As content of rice plants due to changing the redox status of the soil. Phosphate and silicon transporters can be used by As to enter the rice root cells, therefore detoxification mechanisms of As in rice greatly depend on the activity of these transporters. In this review, we covered the main factors that affect the uptake, accumulation, and translocation of As in different plant organs in rice. We investigated the different soil factors and plant cell transporters needed to understand the mechanisms. This study may be useful for further research to develop strategies that inhibit As entry and transport in plant cells and contribute to safe food production.

The art of taking sides: Key determinants for the polar localization of silicon transporter.

The art of taking sides: Key determinants for the polar localization of silicon transporter.

Polar localization of a rice silicon transporter requires isoleucine at both C- and N-termini as well as positively charged residues.

Silicon (Si) is important for stable and high yields in rice (Oryza sativa), a typical Si hyperaccumulator. The high Si accumulation is achieved by the cooperation of 2 Si transporters, LOW SILICON 1 (OsLsi1) and OsLsi2, which are polarly localized in cells of the root exodermis and endodermis. However, the mechanism underlying their polar localization is unknown. Here, we identified amino acid residues critical for the polar localization of OsLsi1. Deletion of both N- and C-terminal regions resulted in the loss of its polar localization. Furthermore, the deletion of the C-terminus inhibited its trafficking from the endoplasmic reticulum to the plasma membrane. Detailed site-directed mutagenesis analysis showed that Ile18 at the N-terminal region and Ile285 at the C-terminal region were essential for the polar localization of OsLsi1. Moreover, a cluster of positively charged residues at the C-terminal region is also required for polar localization. Phosphorylation and Lys modifications of OsLsi1 are unlikely to be involved in its polar localization. Finally, we showed that the polar localization of OsLsi1 is required for the efficient uptake of Si. Our study not only identified critical residues required for the polar localization of OsLsi1, but also provided experimental evidence for the importance of transporter polarity for efficient nutrient uptake.

Genome-wide identification and functional analysis of silicon transporter family genes in moso bamboo (Phyllostachys edulis)

Silicon (Si) has crucial effects on plant development and stress resistance. Silicon transporters regulate Si absorption, transport, and distribution in plants. In this study, we identified and characterized the Si transporter gene family of moso bamboo (Phyllostachys edulis) and cloned seven putative Si transporter genes. Moso bamboo Si transporters contain conserved functional domains that mediate the accumulation of considerable amounts of Si. The analysis of gene duplication patterns and divergence times suggested that the expansion of the moso bamboo Si transporter family was mainly due to segmental duplications. The expression of moso bamboo Si transporter genes, which varied among organs, was significantly modulated by Si treatments. The subcellular localization analysis showed that Si transporters are plasma membrane proteins. The Si content increased in transgenic Arabidopsis overexpressing PeLsi1-1 or PeLsi1-2, which affected vegetative and reproductive growth. Our single-particle tracking analysis revealed the four diffusion modes of PeLsi1-1 on the plasma membrane. Moreover, the particle velocity, dwell time, and motion range of PeLsi1-1 decreased in response to Si treatments. The results of this study will further clarify the molecular mechanisms underlying Si absorption and accumulation in bamboo plants.

Computational modelling of diatom silicic acid transporters predicts a conserved fold with implications for their function and evolution

Diatoms are an important group of algae that can produce intricate silicified cell walls (frustules). The complex process of silicification involves a set of enigmatic integral membrane proteins that are thought to actively transport the soluble precursor of biosilica, dissolved silicic acid. Full-length silicic acid transporters are found widely across the diatoms while homologous shorter proteins have now been identified in a range of other organisms. It has been suggested that modern silicic acid transporters arose from the union of such partial sequences. Here, we present a computational study of the silicic acid transporters and related transporter-like sequences to help understand the structure, function and evolution of this class of membrane protein. The AlphaFold software predicts that all of the protein sequences studied here share a common fold in the membrane domain which is entirely different from the predicted folds of non-homologous silicic acid transporters from plants. Substrate docking reveals how conserved polar residues could interact with silicic acid at a central solvent-accessible binding site, consistent with an alternating access mechanism of transport. The structural conservation between these proteins supports a model where modern silicon transporters evolved from smaller ancestral proteins by gene fusion.

Genome-wide identification and functional prediction of silicon (Si) transporters in poplar (Populus trichocarpa)

Silicon (Si) enhances plant tolerance to various biotic and abiotic stressors such as salinity, drought, and heat. In addition, Si can be biomineralized within plants to form organic carbon-containing phytoliths that can have ecosystem-level consequences by contributing to long-term carbon sequestration. Si is taken up and transported in plants via different transporter proteins such as influx transporters (e.g., Lsi1, Lsi6) and efflux transporters (e.g., Lsi2). Additionally, the imported Si can be deposited in plant leaves via silicification process using the Siliplant 1 (e.g., Slp1) protein. Functional homologs of these proteins have been reported in different food crops. Here, we performed a genome-wide analysis to identify different Si transporters and Slp1 homologs in the bioenergy crop poplar (Populus trichocarpa Torr. and A. Gray ex W. Hook). We identified one channel-type Si influx transporter (PtLsi1; Potri.017G083300), one Si efflux transporter (PtLsi2; Potri.012G144000) and two proteins like Slp1 (PtSlp1a; Potri.004G168600 and PtSlp1b; Potri.009G129900) in the P. trichocarpa genome. We found a unique sequence (KPKPPVFKPPPVPI) in PtSlp1a which is repeated six times. Repeated presence of this sequence in PtSlp1a indicates that this protein might be important for silicification processes in P. trichocarpa. The mutation profiles of different Si transporters in a P. trichocarpa genome-wide association study population identified significant and impactful mutations in Potri.004G168600 and Potri.009G129900. Using a publically accessible database (http://bar.utoronto.ca/eplant_poplar/), digital expression analysis of the putative Si transporters in P. trichocarpa found low to moderate expression in the anticipated tissues, such as roots and leaves. Subcellular localization analysis found that PtLsi1/PtLsi2 are localized in the plasma membrane, whereas PtSlp1a/PtSlp1b are found in the extracellular spaces. Protein–Protein interaction analysis of PtLsi1/PtLsi2 identified Delta-1-pyrroline-5-carboxylate synthase (P5CS) as one of the main interacting partners of PtLsi2, which plays a key role in proline biosynthesis. Proline is a well-known participant in biotic and abiotic stress tolerance in plants. These findings will reinforce future efforts to modify Si accumulation for enhancing plant stress tolerance and carbon sequestration in poplar.