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1.
Promoter analysis of the barley Pht1;1 phosphate transporter gene identifies regions controlling root expression and responsiveness to phosphate deprivation
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Previous studies have shown that the promoter from the barley (Hordeum vulgare) phosphate transporter gene, HvPht1;1, activates high levels of expression in rice (Oryza sativa) roots and that the expression level was induced by up to 4-fold in response to phosphorus (P) deprivation. To identify promoter regions controlling gene regulation specificities, successive promoter truncations were made and attached to reporter genes. Promoters of between 856 and 1,400 nucleotides activated gene expression in a number of cell types but with maximal expression in trichoblast (root hair) cells. For shorter promoters the trichoblast specificity was lost, but in other tissues the distribution pattern was unchanged. The low P induction response was unaffected by promoter length. Domain exchange experiments subsequently identified that the region between -856 and -547 nucleotides (relative to the translational start) is required for epidermal cell expression. A second region located between 0 and -195 nucleotides controls root-tip expression. The HvPht1;1 promoter contains one PHO-like motif and three motifs similar to the dicot P1BS element. Analysis of promoters from which the PHO-like element was eliminated (by truncation) showed no change in the gene induction response to P deficiency. In contrast, mutation of the P1BS elements eliminated any induction of gene expression in response to low P. An internal HvPht1;1 promoter fragment, incorporating a single P1BS element, had an increased response to P deprivation in comparison with the unmodified promoter (containing three elements). Together these findings further our understanding of the regulation of the HvPht1;1 gene and provide direct evidence for a functional role of the P1BS element in the expression of P-regulated genes. 相似文献
2.
Expression analysis suggests novel roles for members of the Pht1 family of phosphate transporters in Arabidopsis 总被引:14,自引:0,他引:14
Mudge SR Rae AL Diatloff E Smith FW 《The Plant journal : for cell and molecular biology》2002,31(3):341-353
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. 相似文献
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Promoters of phosphate transporter genes MtPT1 and MtPT2 of Medicago truncatula were isolated by utilizing the gene-space sequence information and by screening of a genomic library, respectively. Two reporter genes, beta-glucuronidase (GUS) and green fluorescent protein (GFP) were placed under the control of the MtPT1 and MtPT2 promoters. These chimeric transgenes were introduced into Arabidopsis thaliana and transgenic roots of M. truncatula, and expression patterns of the reporter genes were assayed in plants grown under different phosphate (Pi) concentrations. The expression of GUS and GFP was only observed in root tissues, and the levels of expression decreased with increasing concentrations of Pi. GUS activities in roots of transgenic plants decreased 10-fold when the plants were transferred from 10 microM to 2 mM Pi conditions, however, when the plants were transferred back to 10 microM Pi conditions, GUS expression reversed back to the original level. The two promoters lead to different expression patterns inside root tissues. The MtPT1 promoter leads to preferential expression in root epidermal and cortex cells, while MtPT2 promoter results in strong expression in the vascular cylinder in the center of roots. Promoter deletion analyses revealed possible sequences involved in root specificity and Pi responsiveness. The promoters are valuable tools for defined engineering of plants, particularly for root-specific expression of transgenes. 相似文献
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OsCSLD1, a cellulose synthase-like D1 gene, is required for root hair morphogenesis in rice 总被引:3,自引:0,他引:3
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Kim CM Park SH Je BI Park SH Park SJ Piao HL Eun MY Dolan L Han CD 《Plant physiology》2007,143(3):1220-1230
Root hairs are long tubular outgrowths that form on the surface of specialized epidermal cells. They are required for nutrient and water uptake and interact with the soil microflora. Here we show that the Oryza sativa cellulose synthase-like D1 (OsCSLD1) gene is required for root hair development, as rice (Oryza sativa) mutants that lack OsCSLD1 function develop abnormal root hairs. In these mutants, while hair development is initiated normally, the hairs elongate less than the wild-type hairs and they have kinks and swellings along their length. Because the csld1 mutants develop the same density and number of root hairs along their seminal root as the wild-type plants, we propose that OsCSLD1 function is required for hair elongation but not initiation. Both gene trap expression pattern and in situ hybridization analyses indicate that OsCSLD1 is expressed in only root hair cells. Furthermore, OsCSLD1 is the only member of the four rice CSLD genes that shows root-specific expression. Given that the Arabidopsis (Arabidopsis thaliana) gene KOJAK/AtCSLD3 is required for root hair elongation and is expressed in the root hair, it appears that OsCSLD1 may be the functional ortholog of KOJAK/AtCSLD3 and that these two genes represent the root hair-specific members of this family of proteins. Thus, at least part of the mechanism of root hair morphogenesis in Arabidopsis is conserved in rice. 相似文献
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Cereal phosphate transporters associated with the mycorrhizal pathway of phosphate uptake into roots 总被引:1,自引:0,他引:1
A very large number of plant species are capable of forming symbiotic associations with arbuscular mycorrhizal (AM) fungi.
The roots of these plants are potentially capable of absorbing P from the soil solution both directly through root epidermis
and root hairs, and via the AM fungal pathway that delivers P to the root cortex. A large number of phosphate (P) transporters
have been identified in plants; tissue expression patterns and kinetic information supports the roles of some of these in
the direct root uptake pathways. Recent work has identified additional P transporters in several unrelated species that are
strongly induced, sometimes specifically, in AM roots. The primary aim of the work described in this paper was to determine
how mycorrhizal colonisation by different species of AM fungi influenced the expression of members of the Pht1 gene families in the cereals Hordeum vulgare (barley), Triticum aestivum (wheat) and Zea mays (maize). RT-PCR and in-situ hybridisation, showed that the transporters HORvu;Pht1;8 (AY187023), TRIae;Pht1;myc (AJ830009) and ZEAma;Pht1;6 (AJ830010), had increased expression in roots colonised by the AM fungi Glomus intraradices,Glomus sp. WFVAM23 and Scutellospora calospora. These findings add to the increasing body of evidence indicating that plants that form AM associations with members of the
Glomeromycota have evolved phosphate transporters that are either specifically or preferentially involved in scavenging phosphate
from the apoplast between intracellular AM structures and root cortical cells. Operation of mycorrhiza-inducible P transporters
in the AM P uptake pathway appears, at least partially, to replace uptake via different P transporters located in root epidermis
and root hairs.
Electronic Supplementary Material Supplementary material is available for this article at 相似文献
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Ton Gloudemans T. V. Bhuvaneswari Marja Moerman Ton van Brussel Albert van Kammen Ton Bisseling 《Plant molecular biology》1989,12(2):157-167
The mRNA population in pea root hairs was characterized by means of in vitro translation of total root hair RNA followed by 2-dimensional gel electrophoresis of the translation products. Root hairs contain several mRNAs not detectable in total RNA preparations from roots. Most of these root hair-specific mRNAs occur in elongating root hairs at higher levels than in mature root hairs. The expression of some genes in pea root hairs is typically affected by inoculation with Rhizobium leguminosarum. One gene, encoding RH-42, is specifically induced while the expression of another gene, encoding RH-44, is markedly enhanced. Using R. leguminosarum mutants it was shown that the nodC gene is required for the induction and enhancement of expression of the RH-42 and RH-44 genes, respectively, while the Rhizobium chromosomal gene pss1, involved in exopolysaccharide synthesis, is not essential. After induction of the nod genes with apigenin the bacteria excrete into the culture medium a factor that causes root hair deformation. This deformation factor stimulates the expression of the RH-44 gene but does not induce the expression of the gene encoding RH-42. 相似文献
10.
The phosphate uptake mechanism 总被引:11,自引:0,他引:11
Frank W. Smith 《Plant and Soil》2002,245(1):105-114
The slow rate of diffusion of phosphate in soil results in a zone of depletion of phosphate ions in solution around the roots of plants in low phosphate soils. Transfer of phosphate to the site of uptake into the root symplasm limits phosphate uptake in such soils. This transfer involves movement across the depletion zone and through the root apoplasm. The apoplasm is made up of the cell walls of epidermal and cortical cells, together with the associated intercellular spaces. Although the pores in the open latticework of these cell walls permit movement of nutrients around cells, they increase the path length across which phosphate ions have to diffuse. The structural components and net negative charges of the cell walls also influence the effective concentrations of phosphate in the apoplasm. This concentration may be further modified by excreted organic compounds around cell walls and the presence of micro-organisms that use such compounds as carbon sources. A membrane on the inner surface of the cell wall, the plasmalemma, separates the apoplasm from the symplasm. Uptake of nutrients into the root symplasm occurs through transporter proteins embedded in this membrane. Understanding of the mechanisms by which phosphate is transported across the plasmalemma into the plant symplasm has advanced considerably over the past 4 years due to the application of molecular techniques. Genes encoding the transporters involved in this process have been isolated from a number of plant species. These transporters belong to a family of membrane proteins characterized by having 12 membrane-spanning domains arranged in a '6+6' configuration. H2PO4
– ions, together with protons, are transported through this protein. This transport process is driven by the potential across the membrane maintained by the action of a H+-ATPase, the `proton pump', that extrudes protons to the outer surface of the membrane. The expression of genes encoding high-affinity root phosphate transporters is regulated by the phosphorus (P) status of the plant. The transduction pathway involved in this regulation is not known at present. It is a systemic response rather than a localized response, however, the overall phosphate status of the plant being the controlling factor. Under phosphate stress, the expression of genes encoding these phosphate transporters is up-regulated. This results in a greater number of transporter proteins in the plasmalemma and enhanced phosphate uptake rates, if phosphate is available at the membrane surface. Uptake occurs around the root tip, into epidermal cells with their associated root hairs and into cells in the outer layers of the root cortex. Further back along the root axis, phosphate can also be taken up by transfer from mycorrhizal fungi to root cortical cells.Strategies for increasing nutrient uptake by overexpressing genes encoding high-affinity phosphate transporters are likely to be mainly applicable to situations where a reasonable phosphate concentration can be maintained at the outer surface of the plasmalemma. Maintaining such a concentration is a major problem in the phosphate deficient soils of the semi-arid tropics (SAT), so emphasis in these soils is on strategies to improve the movement of phosphate to the surface of the plasmalemma. There may be scope, however, for manipulating the expression of genes involved in the internal mobilisation of phosphate within the plant, thereby improving phosphate utilisation. 相似文献
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Summary Measurements were made of the diffusion of P32-labelled phosphate to single roots of onion, leek and rye-grass growing in an Upper Greensand sandy loam (UGS) and a Coral
Rag Clay (CRC) to which different amounts of phosphate had been added. Concentration-dependent diffusion coefficients for
phosphate ions in the soils were calculated from phosphate desorption isotherms in calcium chloride. The experimental uptake
by roots of known dimensions was compared with supply expected by diffusion to a cylindrical model root of the same dimensions.
Allowance was made for absorption by the root hairs on rye-grass roots.
Phosphate absorption by a cm length of intact root was found to continue for at least 16 days for onion, 10 days for leek
and 5 days for rye-grass. Over a wide range of conditions (phosphate concentrations, soils, plant species), experimental uptake
was close to the maximum calculated to be possible for the diffusion model except on one soil at a high level of phosphate.
Although the concentration of phosphate in the soil solution at the root boundary appeared to be reduced to a small fraction
of the initial concentration, because of the extreme non-linear form of the desorption isotherm less than 1/2 of the P32 exchangeable pool of P was considered to contribute to diffusion.
Phosphate uptake by rye grass could only be accounted for if the root hairs were active. Although only a small fraction of
the uptake is derived from inside the root hair cylinder, this increases the efficiency of the central root 2.3 fold by providing
a zone close to the central root through which phosphate moves very readily. 相似文献
13.
The molecular physiology of ammonium uptake and retrieval 总被引:18,自引:0,他引:18
Plants are able to take up ammonium from the soil, or through symbiotic interactions with microorganisms, via the root system. Using functional complementation of yeast mutants, it has been possible to identify a new class of membrane proteins, the ammonium transporter/methylammonium permease (AMT/MEP) family, that mediate secondary active ammonium uptake in eukaryotic and prokaryotic organisms. In plants, the AMT gene family can be subdivided according to their amino-acid sequences into three subfamilies: a large subfamily of AMT1 genes and two additional subfamilies each with single members (LeAMT1;3 from tomato and AtAMT2;1 from Arabidopsis thaliana). These transporters vary especially in their kinetic properties and regulatory mechanism. High-affinity transporters are induced in nitrogen-starved roots, whereas other transporters may be considered as the 'work horses' that are active when conditions are conducive to ammonium assimilation. The expression of several AMTs in root hairs further supports a role in nutrient acquisition. These studies provide basic information that will be needed for the dissection of nitrogen uptake by plants at the molecular level and for determining the role of individual AMTs in nutrient uptake and potentially in nutrient efficiency. 相似文献
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Regulation of sulfate uptake and expression of sulfate transporter genes in Brassica oleracea as affected by atmospheric H(2)S and pedospheric sulfate nutrition
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Buchner P Stuiver CE Westerman S Wirtz M Hell R Hawkesford MJ De Kok LJ 《Plant physiology》2004,136(2):3396-3408
Demand-driven signaling will contribute to regulation of sulfur acquisition and distribution within the plant. To investigate the regulatory mechanisms pedospheric sulfate and atmospheric H(2)S supply were manipulated in Brassica oleracea. Sulfate deprivation of B. oleracea seedlings induced a rapid increase of the sulfate uptake capacity by the roots, accompanied by an increased expression of genes encoding specific sulfate transporters in roots and other plant parts. More prolonged sulfate deprivation resulted in an altered shoot-root partitioning of biomass in favor of the root. B. oleracea was able to utilize atmospheric H(2)S as S-source; however, root proliferation and increased sulfate transporter expression occurred as in S-deficient plants. It was evident that in B. oleracea there was a poor shoot to root signaling for the regulation of sulfate uptake and expression of the sulfate transporters. cDNAs corresponding to 12 different sulfate transporter genes representing the complete gene family were isolated from Brassica napus and B. oleracea species. The sequence analysis classified the Brassica sulfate transporter genes into four different groups. The expression of the different sulfate transporters showed a complex pattern of tissue specificity and regulation by sulfur nutritional status. The sulfate transporter genes of Groups 1, 2, and 4 were induced or up-regulated under sulfate deprivation, although the expression of Group 3 sulfate transporters was not affected by the sulfate status. The significance of sulfate, thiols, and O-acetylserine as possible signal compounds in the regulation of the sulfate uptake and expression of the transporter genes is evaluated. 相似文献
18.
This paper reports morphological and physiological characteristics of a first root-hairless mutant (RH2) of rice (Oryza sativa L.), which can be useful in advancing knowledge on the role of root hairs in water and nutrient uptake, and genetics of root hairs. The mutant was selected among NaN3 mutagenized progeny of the rice cultivar Oochikara. Microscopic observations showed absence of root hairs in RH2. At the seedling stage, RH2 showed shorter seedling height and shorter roots compared to the wild type variety Oochikara. Because of the differences in seedling growth, all comparisons between Oochikara and RH2 in uptake-related characters were made on the basis of values adjusted by the dry weight of either the shoot or the root. When grown at low water potential in soil, Oochikara and RH2 were similar in shoot water content and transpiration per unit shoot dry weight, and similarly, at low water potential in solution culture, there was no significant difference between Oochikara and RH2 in transpiration per unit shoot dry weight. These results suggest that at the seedling stage, root hairs do not significantly contribute to uptake of water. In solution culture, Oochikara and RH2 did not significantly differ in phosphate uptake per unit root dry weight. This result supports the previous work that root hairs do not contribute to phosphate uptake in solution culture. Regarding to response to plant hormones, RH2 showed a higher level of resistance to two synthetic auxins, 2,4-dichlorophenoxyacetic acid (2,4-D) and 1-naphthaleneacetic acid (NAA) than Oochikara. NAA treatment induced very short root hairs in RH2, suggesting that the absence of root hairs in RH2 may be due to a shortage of endogenous auxin. Genetic analysis showed that the root hairless character in RH2 is inherited as a single recessive gene. 相似文献
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The storage root (taproot) of sugar beet (Beta vulgaris L.) originates from hypocotyl and primary root and contains many different tissues such as central xylem, primary and secondary cambium, secondary xylem and phloem, and parenchyma. It was the aim of this work to characterize the promoters of three taproot-expressed genes with respect to their tissue specificity. To investigate this, promoters for the genes Tlp, His1-r, and Mll were cloned from sugar beet, linked to reporter genes and transformed into sugar beet and tobacco. Reporter gene expression analysis in transgenic sugar beet plants revealed that all three promoters are active in the storage root. Expression in storage root tissues is either restricted to the vascular zone (Tlp, His1-r) or is observed in the whole organ (Mll). The Mll gene is highly organ specific throughout different developmental stages of the sugar beet. In tobacco, the Tlp and Mll promoters drive reporter gene expression preferentially in hypocotyl and roots. The properties of the Mll promoter may be advantageous for the modification of sucrose metabolism in storage roots. 相似文献
20.
Nitrate induction of root hair density is mediated by TGA1/TGA4 and CPC transcription factors in Arabidopsis thaliana
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Javier Canales Orlando Contreras‐López José M. Álvarez Rodrigo A. Gutiérrez 《The Plant journal : for cell and molecular biology》2017,92(2):305-316
Root hairs are specialized cells that are important for nutrient uptake. It is well established that nutrients such as phosphate have a great influence on root hair development in many plant species. Here we investigated the role of nitrate on root hair development at a physiological and molecular level. We showed that nitrate increases root hair density in Arabidopsis thaliana. We found that two different root hair defective mutants have significantly less nitrate than wild‐type plants, suggesting that in A. thaliana root hairs have an important role in the capacity to acquire nitrate. Nitrate reductase‐null mutants exhibited nitrate‐dependent root hair phenotypes comparable with wild‐type plants, indicating that nitrate is the signal that leads to increased formation of root hairs. We examined the role of two key regulators of root hair cell fate, CPC and WER, in response to nitrate treatments. Phenotypic analyses of these mutants showed that CPC is essential for nitrate‐induced responses of root hair development. Moreover, we showed that NRT1.1 and TGA1/TGA4 are required for pathways that induce root hair development by suppression of longitudinal elongation of trichoblast cells in response to nitrate treatments. Our results prompted a model where nitrate signaling via TGA1/TGA4 directly regulates the CPC root hair cell fate specification gene to increase formation of root hairs in A. thaliana. 相似文献