Screening for Sugarcane Root Phenes Reveals That Reducing Tillering Does Not Lead to an Increased Root Mass Fraction.

Sugarcane root systems are poorly studied and understood due to the perennial nature, tall stature, and the long cropping cycle. Whilst some field studies gave insights into sugarcane root traits, there is no detailed description of root and root system traits available. The objectives of our work were to establish a baseline of sugarcane root trait values that will serve for future studies, and to characterize the degree of root system resilience when restricting tiller number. We first conducted an initial screening for root trait diversity on a collection of twenty cultivars representative of sugarcane breeding from 1930 to now. Then we investigated the effect of reduced tillering, via manual de-tillering, on the plant root and root system traits of five varieties grown under optimal conditions in a glasshouse for 1700°Cd. In addition to establishing baseline means and variation for sugarcane root trait values that could serve as a reference for crop models, we demonstrated that the sugarcane root mass fraction was extremely resilient to drastic reduction in tiller number. Restricted plants were effectively maintaining their root system configuration (opening angle) by dramatically increasing the number of nodal roots produced per tiller as well as maximizing total root length by increasing the specific root length. Using this knowledge of sugarcane root traits in combination with the specific agronomic constraints for sugarcane will now underpin the development of a root system ideotype for sugarcane to enable targeted root trait selection for improving crop productivity.

Overexpression of TaCML20, a calmodulin-like gene, enhances water soluble carbohydrate accumulation and yield in wheat.

Calcium (Ca2+ ) is a universal messenger that mediates intracellular responses to extracellular stimuli in living organisms. Calmodulin (CaM) and calmodulin-like (CML) proteins are the important Ca2+ sensors in plants that decode Ca2+ -signatures to execute downstream intracellular level responses. Several studies indicate the interlinking of Ca2+ and sugar signaling in plants; however, no genes have been functionally characterized to provide molecular evidence. Our study found that expression of TaCML20 was significantly correlated with water soluble carbohydrate (WSC) concentrations in recombinant inbred lines in wheat. TaCML20 has four EF-hand motifs that may facilitate the binding of Ca2+ . To explore the role of CML20, we generated TaCML20 overexpressing transgenic lines in wheat. These lines accumulated higher WSC concentrations in the shoots, and we also found a significantly increased transcript level of sucrose:sucrose-1-fructosyltransferase (1-SST) in the internodes compared with the control plants. In addition, TaCML20 overexpressing plants showed significantly increased tillers per plant and also increased about 19% of grain weight per plant compared with control plants. The results also suggested a role for TaCML20 in drought stress, as its transcripts significantly increased in the shoots of wild-type plants under water deficit. These results uncovered the role of CML20 in determining multiple traits in wheat.

Expression of sugarcane genes associated with perception of photoperiod and floral induction reveals cycling over a 24-hour period.

The genetic network resulting in the production of an inflorescence is complex, involving one or more pathways including the photoperiod, maturity, gibberellin and autonomous pathways, and induction and repression of genes along the pathways. Understanding the cyclic expression profile of genes involved with photoperiod perception and floral pathway induction in sugarcane, an intermediate-short day plant (ISD), is crucial for identifying key genes and understanding how the profile changes in response to floral induction signals under decreasing daylengths. Homologues of 21 genes, and some gene alleles, associated with photoperiod perception and the flower induction pathway were examined in sugarcane variety Q174 over a 24-h light-dark cycle. The strongest expression of these genes was seen in the immature spindle leaves and levels of expression generally decreased with increasing leaf age. Significant changes in gene expression levels during a 24-h cycle were observed for 16 of the 21 genes tested. We have now defined an important baseline for expression patterns over a 24-h cycle in non-inductive conditions in sugarcane. These results can be utilised to select the optimal time for detecting changes during floral induction, differences between varieties that are responsive/non-responsive to photoperiod induction, and to identify genes that may be manipulated to enhance or inhibit flowering.

Identification and Functional Characterization of Sugarcane Invertase Inhibitor (ShINH1): A Potential Candidate for Reducing Pre- and Post-harvest Loss of Sucrose in Sugarcane.

In sugarcane, invertase enzymes play a key role in sucrose accumulation and are also involved in futile reactions where sucrose is continuously degraded during the pre- and post-harvest period, thereby reducing sugar yield and recovery. Invertase inhibitor (INVINH) proteins play a key role in post-translation regulation of plant invertases through which sucrose hydrolysis is controlled. INVINH proteins are small (18 kDa) members of the pectin methylesterase inhibitor superfamily and they are moderately conserved across plants. In the present study, we identified two INVINH genes from sugarcane, ShINH1 and ShINH2. In silico characterization of the encoded proteins revealed 43% sequence identity at the amino acid level, confirming the non-allelic nature of the proteins. The presence of putative signal peptide and subcellular targeting sequences revealed that ShINH1 and ShINH2 likely have apoplasmic and vacuolar localization, respectively. Experimental visualization of ShINH1-GFP revealed that ShINHI is indeed exported to the apoplast. Differential tissue-specific and developmental expression of ShINH1 between leaf, stalk, flower and root suggest that it plays a role in controlling source-sink metabolic regulation during sucrose accumulation in sugarcane. ShINH1 is expressed at relatively high levels in leaves and stalk compared to flowers and roots, and expression decreases significantly toward internodal maturity during stalk development. ShINH1 is expressed at variable levels in flowers with no specific association to floral maturity. Production of recombinant ShINH1 enabled experimental validation of protein function under in vitro conditions. Recombinant ShINH1 potently inhibited acid invertase (IC50 22.5 nM), making it a candidate for controlling pre- and post-harvest deterioration of sucrose in sugarcane. Our results indicate that ShINH1 and ShINH2 are likely to play a regulatory role in sucrose accumulation and contribute to the improvement of sugar yield and recovery in sugarcane.

An analysis of the role of the ShSUT1 sucrose transporter in sugarcane using RNAi suppression.

The role of ShSUT1 in sucrose mobilisation and storage in sugarcane was investigated by employing RNAi technology to reduce the expression of this gene. Transcript profiling in non-transformed plants showed an alignment between expression and sucrose concentration, with strongest expression in source leaves and increasing expression through the daylight period of a diurnal cycle. Five transgenic plant lines were produced with reduced ShSUT1 expression ranging from 52 to 92% lower than control plants. Differential suppression of ShSUT1 sequence variants in the highly polyploid sugarcane genome were also investigated. Amplicon sequencing of the ShSUT1 variants within the transgenic lines and controls showed no preferential suppression with only minor differences in the proportional expression of the variants. A range of altered sugar, fibre and moisture contents were measured in mature leaf and internode samples, but no phenotype was consistently exhibited by all five transgenic lines. Phenotypes observed indicate that ShSUT1 does not play a direct role in phloem loading. ShSUT1 is likely involved with retrieving sucrose from intercellular spaces for transport and storage.

Sucrose Transporter Localization and Function in Phloem Unloading in Developing Stems.

How sucrose transporters (SUTs) regulate phloem unloading in monocot stems is poorly understood and particularly so for species storing high Suc concentrations. To this end, Sorghum bicolor SUTs SbSUT1 and SbSUT5 were characterized by determining their transport properties heterologously expressed in yeast or Xenopus laevis oocytes, and their in planta cellular and subcellular localization. The plasma membrane-localized SbSUT1 and SbSUT5 exhibited a strong selectivity for Suc and high Suc affinities in X. laevis oocytes at pH 5-SbSUT1, 6.3 ± 0.7 mm, and SbSUT5, 2.4 ± 0.5 mm Suc. The Suc affinity of SbSUT1 was dependent on membrane potential and pH. In contrast, SbSUT5 Suc affinity was independent of membrane potential and pH but supported high transport rates at neutral pH. Suc transport by the tonoplast localized SbSUT4 could not be detected using yeast or X. laevis oocytes. Across internode development, SUTs, other than SbSUT4, were immunolocalized to sieve elements, while for elongating and recently elongated internodes, SUTs also were detected in storage parenchyma cells. We conclude that apoplasmic Suc unloading from de-energized protophloem sieve elements in meristematic zones may be mediated by reversal of SbSUT1 and/or by uniporting SWEETs. Storage parenchyma localized SbSUT1 and SbSUT5 may accumulate Suc from the stem apoplasms of elongating and recently elongated internodes, whereas SbSUT4 may function to release Suc from vacuoles. Transiting from an apoplasmic to symplasmic unloading pathway as the stem matures, SbSUT1 and SbSUT5 increasingly function in Suc retrieval into metaphloem sieve elements to maintain a high turgor to drive symplasmic unloading by bulk flow.

Abiotic stress upregulated TaZFP34 represses the expression of type-B response regulator and SHY2 genes and enhances root to shoot ratio in wheat.

Q-type C2H2 zinc finger proteins (ZFPs) are plant-specific DNA-binding proteins containing a conserved QALGGH motif. This study investigated the function of abiotic stress-inducible and predominantly root-expressed Triticum aestivum ZFPs (TaZFP22, TaZFP34 and TaZFP46) with a focus on TaZFP34. Expression of TaZFP34 in roots was upregulated by high salinity, dehydration, oxidative and cold stresses. Overexpression of TaZFP34 in wheat roots resulted in an increased root-to-shoot ratio, a phenomenon observed during plant adaptation to drying soil. Expression of a number of genes which are potentially involved in modulating root growth was significantly altered in the roots of TaZFP34 overexpressing lines. In particular, the transcript levels of TaRR12B, TaRR12D and TaSHY2 that are homologues of known negative regulators of root growth were significantly reduced. Expression of shoot growth-related genes, such as GA3-ox and expansins, was downregulated in the transgenic shoots. TaZFP34 bound to (C/G)AGT(G/A)-like elements in the promoters of TaZFP34 down-regulated TaRR12D and TaSHY2 and transrepressed the reporter gene expression driven by TaRR12D and TaSHY2 promoters. Expression of the above reporter genes was also repressed by TaZFP46 and TaZFP22. These data suggest that TaZFP34 is a transcriptional repressor and is involved in modulating the root-to-shoot ratio.

Drought-Up-Regulated TaNAC69-1 is a Transcriptional Repressor of TaSHY2 and TaIAA7, and Enhances Root Length and Biomass in Wheat.

A well-known physiological adaptation process of plants encountering drying soil is to achieve water balance by reducing shoot growth and maintaining or promoting root elongation, but little is known about the molecular basis of this process. This study investigated the role of a drought-up-regulated Triticum aestivum NAC69-1 (TaNAC69-1) in the modulation of root growth in wheat. TaNAC69-1 was predominantly expressed in wheat roots at the early vegetative stage. Overexpression of TaNAC69-1 in wheat roots using OsRSP3 (essentially root-specific) and OsPIP2;3 (root-predominant) promoters resulted in enhanced primary seminal root length and a marked increase in maturity root biomass. Competitive growth analysis under water-limited conditions showed that OsRSP3 promoter-driven TaNAC69-1 transgenic lines produced 32% and 35% more above-ground biomass and grains than wild-type plants, respectively. TaNAC69-1 overexpression in the roots down-regulated the expression of TaSHY2 and TaIAA7, which are from the auxin/IAA (Aux/IAA) transcriptional repressor gene family and are the homologs of negative root growth regulators SHY2/IAA3 and IAA7 in Arabidopsis. The expression of TaSHY2 and TaIAA7 in roots was down-regulated by drought stress and up-regulated by cytokinin treatment, which inhibited root growth. DNA binding and transient expression analyses revealed that TaNAC69-1 bound to the promoters of TaSHY2 and TaIAA7, acted as a transcriptional repressor and repressed the expression of reporter genes driven by the TaSHY2 or TaIAA7 promoter. These data suggest that TaNAC69-1 is a transcriptional repressor of TaSHY2 and TaIAA7 homologous to Arabidopsis negative root growth regulators and is likely to be involved in promoting root elongation in drying soil.

A strong root-specific expression system for stable transgene expression in bread wheat.

KEY MESSAGE: A strong, stable and root-specific expression system was developed from a rice root-specific GLYCINE - RICH PROTEIN 7 promoter for use as an enabling technology for genetic manipulation of wheat root traits. Root systems play an important role in wheat productivity. Genetic manipulation of wheat root traits often requires a root-specific or root-predominant expression system as an essential enabling technology. In this study, we investigated promoters from rice root-specific or root-predominant expressed genes for development of a root expression system in bread wheat. Transient expression analysis using a GREEN FLUORESCENT PROTEIN (GFP) reporter gene driven by rice promoters identified six promoters that were strongly expressed in wheat roots. Extensive organ specificity analysis of three rice promoters in transgenic wheat revealed that the promoter of rice GLYCINE-RICH PROTEIN 7 (OsGRP7) gene conferred a root-specific expression pattern in wheat. Strong GFP fluorescence in the seminal and branch roots of wheat expressing GFP reporter driven by the OsGRP7 promoter was detected in epidermal, cortical and endodermal cells in mature parts of the root. The GFP reporter driven by the promoter of rice METALLOTHIONEIN-LIKE PROTEIN 1 (OsMTL1) gene was mainly expressed in the roots with essentially no expression in the leaf, stem or seed. However, it was also expressed in floral organs including glume, lemma, palea and awn. In contrast, strong expression of rice RCg2 promoter-driven GFP was found in many tissues. The GFP expression driven by these three rice promoters was stable in transgenic wheat plants through three generations (T1-T3) examined. These data suggest that the OsGRP7 promoter can provide a strong, stable and root-specific expression system for use as an enabling technology for genetic manipulation of wheat root traits.

Tissue-specific transcriptome analysis within the maturing sugarcane stalk reveals spatial regulation in the expression of cellulose synthase and sucrose transporter gene families.

Sugarcane (Saccharum spp. hybrids) accumulates high concentrations of sucrose in its mature stalk and a considerable portion of carbohydrate metabolism is also devoted to cell wall synthesis and fibre production. We examined tissue-specific expression patterns to explore the spatial deployment of pathways responsible for sucrose accumulation and fibre synthesis within the stalk. We performed expression profiling of storage parenchyma, vascular bundles and rind dissected from a maturing stalk internode of sugarcane, identifying ten cellulose synthase subunit genes and examining significant differences in the expression of their corresponding transcripts and those of several sugar transporters. These were correlated with differential expression patterns for transcripts of genes encoding COBRA-like proteins and other cell wall metabolism-related proteins. The sugar transporters genes ShPST2a, ShPST2b and ShSUT4 were significantly up-regulated in storage parenchyma while ShSUT1 was up-regulated in vascular bundles. Two co-ordinately expressed groups of cell wall related transcripts were also identified. One group, associated with primary cell wall synthesis (ShCesA1, ShCesA7, ShCesA9 and Shbk2l3), was up-regulated in parenchyma. The other group, associated with secondary cell wall synthesis (ShCesA10, ShCesA11, ShCesA12 and Shbk-2), was up-regulated in rind. In transformed sugarcane plants, the ShCesA7 promoter conferred stable expression of green fluorescent protein preferentially in the storage parenchyma of the maturing stalk internode. Our results indicate that there is spatial separation for elevated expression of these important targets in both sucrose accumulation and cell wall synthesis, allowing for increased clarity in our understanding of sucrose transport and fibre synthesis in sugarcane.

Poor Fertility, Short Longevity, and Low Abundance in the Soil Seed Bank Limit Volunteer Sugarcane from Seed.

The recent development of genetically modified sugarcane, with the aim of commercial production, requires an understanding of the potential risks of increased weediness of sugarcane as a result of spread and persistence of volunteer sugarcane. As sugarcane is propagated vegetatively from pieces of stalk and the seed plays no part in the production cycle, the fate of seed in the environment is yet to be studied. In this study, sugarcane seed samples, collected in fields over a 2-year period, were used to determine the overall level of sugarcane fertility, seed dormancy, and longevity of seed under field conditions. A survey of the soil seed bank in and around sugarcane fields was used to quantify the presence of sugarcane seeds and to identify and quantify the weeds that would compete with sugarcane seedlings. We demonstrated that under field conditions, sugarcane has low fertility and produces non-dormant seed. The viability of the seeds decayed rapidly (half-life between 1.5 and 2.1 months). This means that, in Australia, sugarcane seeds die before they encounter climatic conditions that could allow them to germinate and establish. Finally, the soil seed bank analysis revealed that there were very few sugarcane seeds relative to the large number of weed seeds that exert a large competitive effect. In conclusion, low fertility, short persistence, and poor ability to compete limit the capacity of sugarcane seed spread and persistence in the environment.

A soluble acid invertase is directed to the vacuole by a signal anchor mechanism.

Enzyme activities in the vacuole have an important impact on the net concentration of sucrose. In sugarcane (Saccharum hybrid), immunolabelling demonstrated that a soluble acid invertase (ß-fructofuranosidase; EC 3.2.1.26) is present in the vacuole of storage parenchyma cells during sucrose accumulation. Examination of sequences from sugarcane, barley and rice showed that the N-terminus of the invertase sequence contains a signal anchor and a tyrosine motif, characteristic of single-pass membrane proteins destined for lysosomal compartments. The N-terminal peptide from the barley invertase was shown to be capable of directing the green fluorescent protein to the vacuole in sugarcane cells. The results suggest that soluble acid invertase is sorted to the vacuole in a membrane-bound form.

Fruit fly bioassay to distinguish "sweet" sugar structures.

Palatable response to dietary sugars plays a significant role in influencing metabolic health. New structures are being explored with beneficial health properties, although consumer acceptance relies heavily on desirable sensory properties. Despite the importance of behavioral responses, the ability to elucidate structure-preference relationships of sugars is lacking. A wild population of Drosophila melanogaster was used as a model to perform pairwise comparisons across structural groups to characterize a fruit fly bioassay for assessing sugar preference. Preference was successfully described in structurally relevant terms, particularly through the ability to directly test sugars of related structures in addition to standard sucrose comparisons. The fruit fly bioassay also provided the first report on the relative preference for the ß-linked sugar alcohol, gentiobiitol. In making reference to well-known human preferences, the bioassay also raises opportunities for greater understanding of behavioral response to sugar structures in general.

A bioinformatic approach to the identification of a conserved domain in a sugarcane legumain that directs GFP to the lytic vacuole.

Sugarcane is an ideal candidate as a biofactory for the production of alternate higher value products. One way of achieving this is to direct useful proteins into the vacuoles within the sugarcane storage parenchyma tissue. By bioinformatic analysis of gene sequences from putative sugarcane vacuolar proteins a motif has been identified that displays high conservation across plant legumain homologues that are known to function within vacuolar compartments. This five amino acid motif, represented by the sequence IRLPS in sugarcane is shown to direct an otherwise secreted GFP fusion protein into a large acidic and proteolytic vacuole in sugarcane callus cells as well as in diverse plant species. In mature sugarcane transgenic plants, the stability of GFP appeared to be dependent on cell type, suggesting that the vacuolar environment can be hostile to introduced proteins. This targeting motif will be a valuable tool for engineering plants such as sugarcane for production of novel products.

Sucrose partitioning between vascular bundles and storage parenchyma in the sugarcane stem: a potential role for the ShSUT1 sucrose transporter.

A transporter with homology to the SUT/SUC family of plant sucrose transporters was isolated from a sugarcane (Saccharum hybrid) stem cDNA library. The gene, designated ShSUT1, encodes a protein of 517 amino acids, including 12 predicted membrane-spanning domains and a large central cytoplasmic loop. ShSUT1 was demonstrated to be a functional sucrose transporter by expression in yeast. The estimated K(m) for sucrose of the ShSUT1 transporter was 2 mM at pH 5.5. ShSUT1 was expressed predominantly in mature leaves of sugarcane that were exporting sucrose and in stem internodes that were actively accumulating sucrose. Immunolocalization with a ShSUT1-specific antiserum identified the protein in cells at the periphery of the vascular bundles in the stem. These cells became lignified and suberized as stem development proceeded, forming a barrier to apoplasmic solute movement. However, the movement of the tracer dye, carboxyfluorescein from phloem to storage parenchyma cells suggested that symplasmic connections are present. ShSUT1 may have a role in partitioning of sucrose between the vascular tissue and sites of storage in the parenchyma cells of sugarcane stem internodes.

Over-expression of a high-affinity phosphate transporter in transgenic barley plants does not enhance phosphate uptake rates.

Transgenic barley plants that over-express the gene encoding a phosphate transporter were generated and used to test the hypothesis that manipulation of transporters may lead to improved phosphate uptake by plant roots. Replicate T2 seedlings from a homozygous line with a single locus insertion were grown in dilute flow culture. The phosphate contents and uptake rates of these plants were compared with control transgenic and wild-type plants. When external phosphate concentration was maintained at 10 µM, all plants including the transgenic over-expressing line displayed low rates of phosphate uptake and contained high levels of phosphate in the shoot tissue. When external phosphate concentration was maintained at 2 µM, the uptake rates increased to a similar level in all plant lines. Three transgenic over-expressing lines were then grown in soil at a range of phosphate concentrations and the dry weights and total phosphorus contents of the shoots were measured and compared to a transgenic control line. The results showed that over-expression of the gene encoding a phosphate transporter did not improve the uptake of phosphate under any of the conditions tested. Transporter activity is likely to be influenced by post-transcriptional mechanisms and will require further investigation before this strategy can be applied to improving plant nutrition.

Identification of a novel sugar transporter homologue strongly expressed in maturing stem vascular tissues of sugarcane by expressed sequence tag and microarray analysis.

The ability of sugarcane to accumulate sucrose provides an experimental system for the study of gene expression determining carbohydrate partitioning and metabolism. A sequence survey of 7242 ESTs derived from the sucrose-accumulating, maturing stem revealed that transcripts for carbohydrate metabolism gene sequences (CMGs) are relatively rare in this tissue. However, within the CMG group, putative sugar transporter ESTs form one of the most abundant classes observed. A combination of EST analysis and microarray and northern hybridization revealed that one of the putative sugar transporter types, designated PST type 2a, was the most abundant and most strongly differentially expressed CMG in maturing stem tissue. PST type 2a is homologous to members of the major facilitator super-family of transporters, possessing 12 predicted transmembrane domains and a sugar transport conserved domain, interrupted by a large cytoplasmic loop. Its transcript was localized to phloem companion cells and associated parenchyma in maturing stem, suggesting a role in sugar translocation rather than storage. In addition, other categories of CMGs show evidence of coordinated expression, such as enzymes involved in sucrose synthesis and cleavage, and a majority of enzymes involved in glycolysis and the pentose phosphate pathway. This study demonstrates the utility of genomic approaches using large-scale EST acquisition and microarray hybridization techniques for studies of the developmental regulation of metabolic enzymes and potential transporters in sugarcane.

Characterization of two phosphate transporters from barley; evidence for diverse function and kinetic properties among members of the Pht1 family.

Putative phosphate transporters have been identified in a barley (Hordeum vulgare L.) genomic library by their homology to known phosphate transporters from dicot species. The genes designated HORvu;Pht1;1 and HORvu;Pht1;6 encode proteins of 521 and 535 amino acids respectively with 12 predicted membrane-spanning domains and other motifs common to the Phtl family of phosphate transporters. HORvu;Pht1;1 is expressed exclusively in roots and is strongly induced by phosphate deprivation. HORvu;Pht1;6 is expressed in the aerial parts of the plant with strongest expression in old leaves and flag leaves. In situ hybridization showed that HORvu;Pht1;6 is expressed in the phloem of vascular bundles in leaves and ears. In order to study the biochemical properties of HORvu;Pht1;1 and HORvu;Pht1;6, the genes were expressed in transgenic rice (Oryza sativa L.) plants under the control of the rice actin promoter and suspension cell cultures were generated. Cells derived from transgenic plants were able to take up phosphate at a much higher rate than control cells, demonstrating that both genes encode functional phosphate transporters. The estimated Km for phosphate for cells expressing HORvu;Pht1;1 was 9.06 +/- 0.82 microM, which is characteristic of a high-affinity transporter. The rate of phosphate uptake decreased with increasing pH, suggesting that HORvu;Pht1;1 operates as a H+/H2PO4(-) symporter. In contrast, the estimated Km for phosphate for cells expressing HORvu;Pht1;6 was 385 +/- 61 microM, which is characteristic of a low-affinity transporter. Taken together, the results suggest that HORvu;Pht1;1 functions in uptake of phosphate at the root surface, while HORvu;Pht1;6 probably functions in remobilization of stored phosphate from leaves.

Localisation of expression of a high-affinity sulfate transporter in barley roots.

The tissue-specific expression pattern of the gene encoding the high-affinity sulfate transporter, HvST1, was examined in sections of barley (Hordeum vulgare L.) roots by in-situ hybridisation. The results showed expression in all cell layers close to the root tip, with increased expression under conditions of sulfate stress. In mature roots, HvST1 was not expressed in sulfate-sufficient conditions, but under sulfate stress, transcripts were detected in the xylem parenchyma, pericycle and endodermis. This pattern of expression is consistent with a role in sulfate uptake and may indicate that the majority of sulfate uptake occurs close to the root tip. In low-sulfate conditions, HvST1 may have an additional role in scavenging sulfate lost during transport.

Expression analysis suggests novel roles for members of the Pht1 family of phosphate transporters in Arabidopsis.

The completion of the Arabidopsis thaliana genome has revealed that there are nine members of the Pht1 family of phosphate transporters in this species. As a step towards identifying the role of this gene family in phosphorus nutrition, we have isolated the promoter regions from each of these genes, and fused them to the reporter genes beta-glucuronidase and/or green fluorescent protein. These chimeric genes have been introduced into A. thaliana, and reporter gene expression has been assayed in plants grown in soil containing high and low concentrations of inorganic phosphate (Pi). Four of these promoters were found to direct reporter gene expression in the root epidermis, and were induced under conditions of phosphate deprivation in a manner similar to previously characterised Pht1 genes. Other members of this family, however, showed expression in a range of shoot tissues and in pollen grains, which was confirmed by RT-PCR. We also provide evidence that the root epidermally expressed genes are expressed most strongly in trichoblasts, the primary sites for uptake of Pi. These results suggest that this gene family plays a wider role in phosphate uptake and remobilisation throughout the plant than was previously believed.