scholarly journals High-affinity ammonium transport by Arabidopsis thaliana AMT1;4

2021 ◽  
Vol 43 (4) ◽  
Author(s):  
Nino Bindel ◽  
Benjamin Neuhäuser
Keyword(s):  
Arabidopsis Thaliana ◽  
Xenopus Laevis ◽  
Pollen Tube ◽  
Pollen Grains ◽  
Transport Proteins ◽  
Ammonium Transporter ◽  
Ammonium Transport ◽  
High Affinity ◽  
Ammonium Transporters ◽  
Unique Expression Pattern

AbstractIn plants high affinity transport proteins mediate the essential transport of ammonium across membranes. In Arabidopsis thaliana six of these AMmonium Transporters (AMTs) are encoded by the genome. All of them show a unique expression pattern. While most AMTs are highly expressed in the root, AtAMT1;4 expression is limited to the pollen grains and the pollen tube. Here, we addressed the transport characteristics of AtAMT1;4 in the heterologous Xenopus laevis oocytes system. The transport saturated and showed high affinity for ammonium with a Km value lower than 10 µM. Based on our electrophysiological analysis, we classified AtAMT1;4 as a high affinity ammonium transporter.

scholarly journals Electrophysiological characterization of AtAMT1;4, an extraordinarily high affinity ammonium transporter from Arabidopsis thaliana

2019 ◽  
Author(s):  
Nino Bindel ◽  
Benjamin Neuhäuser
Keyword(s):  
Arabidopsis Thaliana ◽  
Pollen Tube ◽  
Pollen Grains ◽  
Transport Proteins ◽  
Ammonium Transporter ◽  
High Affinity ◽  
Electrode Voltage ◽  
Electrophysiological Characterization ◽  
Unique Expression Pattern

In plants high affinity transport proteins mediate the essential transport of ammonium across membranes. In Arabidopsis thaliana six of these AMmonium Transporters (AMTs) are encoded on the genome. All of these show a unique expression pattern. While most AMTs are highly expressed in the root, AtAMT1;4 expression is limited to the pollen grains and the pollen tube. Here, we addressed the transport characteristics of AtAMT1;4 in the heterologous Xenopus laevis oocytes system. Two electrode voltage clamp measurements tagged AtAMT1;4 as an electrogenic high affinity ammonium transporter. The transport was saturable and showed extraordinarily high affinity for ammonium with a Km value lower than 10 µM.

scholarly journals Two Expansin Genes, AtEXPA4 and AtEXPB5, Are Redundantly Required for Pollen Tube Growth and AtEXPA4 Is Involved in Primary Root Elongation in Arabidopsis thaliana

Genes ◽  
2021 ◽  
Vol 12 (2) ◽  
pp. 249
Author(s):  
Weimiao Liu ◽  
Liai Xu ◽  
Hui Lin ◽  
Jiashu Cao
Keyword(s):  
Arabidopsis Thaliana ◽  
Pollen Tube ◽  
Pollen Tube Growth ◽  
Cell Walls ◽  
Root Elongation ◽  
Expression Patterns ◽  
Primary Root ◽  
Pollen Grains ◽  
Tube Growth ◽  
Cell Wall Binding

The growth of plant cells is inseparable from relaxation and expansion of cell walls. Expansins are a class of cell wall binding proteins, which play important roles in the relaxation of cell walls. Although there are many members in expansin gene family, the functions of most expansin genes in plant growth and development are still poorly understood. In this study, the functions of two expansin genes, AtEXPA4 and AtEXPB5 were characterized in Arabidopsis thaliana. AtEXPA4 and AtEXPB5 displayed consistent expression patterns in mature pollen grains and pollen tubes, but AtEXPA4 also showed a high expression level in primary roots. Two single mutants, atexpa4 and atexpb5, showed normal reproductive development, whereas atexpa4atexpb5 double mutant was defective in pollen tube growth. Moreover, AtEXPA4 overexpression enhanced primary root elongation, on the contrary, knocking out AtEXPA4 made the growth of primary root slower. Our results indicated that AtEXPA4 and AtEXPB5 were redundantly involved in pollen tube growth and AtEXPA4 was required for primary root elongation.

scholarly journals Differential Expression of Aspergillus nidulans Ammonium Permease Genes Is Regulated by GATA Transcription Factor AreA

2006 ◽  
Vol 5 (2) ◽  
pp. 226-237 ◽  
Author(s):  
Brendon J. Monahan ◽  
Marion C. Askin ◽  
Michael J. Hynes ◽  
Meryl A. Davis
Keyword(s):  
Aspergillus Nidulans ◽  
Sufficient Conditions ◽  
Optimal Growth ◽  
Ammonium Transporter ◽  
Ammonium Uptake ◽  
Ammonium Transport ◽  
Gata Factor ◽  
High Affinity ◽  
Gata Transcription Factor ◽  
Nitrogen Conditions

ABSTRACT The movement of ammonium across biological membranes is mediated in both prokaryotes and eukaryotes by ammonium transport proteins (AMT/MEP) that constitute a family of related sequences. We have previously identified two ammonium permeases in Aspergillus nidulans, encoded by the meaA and mepA genes. Here we show that meaA is expressed in the presence of ammonium, consistent with the function of MeaA as the main ammonium transporter required for optimal growth on ammonium as a nitrogen source. In contrast, mepA, which encodes a high-affinity ammonium permease, is expressed only under nitrogen-limiting or starvation conditions. We have identified two additional AMT/MEP-like genes in A. nidulans, namely, mepB, which encodes a second high-affinity ammonium transporter expressed only in response to complete nitrogen starvation, and mepC, which is expressed at low levels under all nitrogen conditions. The MepC gene product is more divergent than the other A. nidulans AMT/MEP proteins and is not thought to significantly contribute to ammonium uptake under normal conditions. Remarkably, the expression of each AMT/MEP gene under all nitrogen conditions is regulated by the global nitrogen regulatory GATA factor AreA. Therefore, AreA is also active under nitrogen-sufficient conditions, along with its established role as a transcriptional activator in response to nitrogen limitation.

scholarly journals Differential Activation of Ammonium Transporters During the Accumulation of Ammonia by Colletotrichum gloeosporioides and Its Effect on Appressoria Formation and Pathogenicity

2013 ◽  
Vol 26 (3) ◽  
pp. 345-355 ◽  
Author(s):  
Chen Shnaiderman ◽  
Itay Miyara ◽  
Ilana Kobiler ◽  
Amir Sherman ◽  
Dov Prusky
Keyword(s):  
Gene Disruption ◽  
Colletotrichum Gloeosporioides ◽  
Ammonium Transporter ◽  
Ammonium Transport ◽  
Wild Type ◽  
Ammonia Accumulation ◽  
Ammonium Transporters ◽  
Avocado Fruit ◽  
Appressoria Formation ◽  
Encoding Genes

Ammonium secreted by the post-harvest pathogen Colletotrichum gloeosporioides during host colonization accumulates in the host environment due to enhanced fungal nitrogen metabolism. Two types of ammonium transporter-encoding genes, AMET and MEP, are expressed during pathogenicity. Gene disruption of AMET, a gene modulating ammonia secretion, showed twofold reduced ammonia secretion and 45% less colonization on avocado fruit, suggesting a contribution to pathogenicity. MEPB, a gene modulating ammonium transport, is expressed by C. gloeosporioides during pathogenicity and starvation conditions in culture. Gene disruption of MEPB, the most highly expressed gene of the MEP family, resulted in twofold overexpression of MEPA and MEPC but reduced colonization, suggesting MEPB expression's contribution to pathogenicity. Analysis of internal and external ammonia accumulation by ΔmepB strains in mycelia and germinated spores showed rapid uptake and accumulation, and reduced secretion of ammonia in the mutant versus wild-type (WT) strains. Ammonia uptake by the WT germinating spores but not by the ΔmepB strain with compromised ammonium transport activated cAMP and transcription of PKA subunits PKAR and PKA2. ΔmepB mutants showed 75% less appressorium formation and colonization than the WT, which was partially restored by 10 mM exogenous ammonia. Thus, whereas both AMET and MEPB genes modulate ammonia secretion, only MEPB contributes to ammonia accumulation by mycelia and germinating spores that activate the cAMP pathways, inducing the morphogenetic processes contributing to C. gloeosporioides pathogenicity.

scholarly journals Five Gametophytic Mutations Affecting Pollen Development and Pollen Tube Growth in Arabidopsis thaliana

Genetics ◽  
2001 ◽  
Vol 158 (4) ◽  
pp. 1773-1783 ◽  
Author(s):  
Antonia Procissi ◽  
Solveig de Laissardière ◽  
Madina Férault ◽  
Daniel Vezon ◽  
Georges Pelletier ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Pollen Tube ◽  
Pollen Tube Growth ◽  
Pollen Development ◽  
Male Gametophyte ◽  
Pollen Grains ◽  
Segregation Ratio ◽  
Variable Number ◽  
Tube Growth ◽  
Mutant Lines

Abstract Mutant analysis represents one of the most reliable approaches to identifying genes involved in plant development. The screening of the Versailles collection of Arabidopsis thaliana T-DNA insertion transformants has allowed us to isolate different mutations affecting male gametophytic functions and viability. Among several mutated lines, five have been extensively studied at the genetic, molecular, and cytological levels. For each mutant, several generations of selfing and outcrossing have been carried out, leading to the conclusion that all these mutations are tagged and affect only the male gametophyte. However, only one out of the five mutations is completely penetrant. A variable number of T-DNA copies has integrated in the mutant lines, although all segregate at one mutated locus. Two mutants could be defined as “early mutants”: the mutated genes are presumably expressed during pollen grain maturation and their alteration leads to the production of nonfunctional pollen grains. Two other mutants could be defined as “late mutant” since their pollen is able to germinate but pollen tube growth is highly disturbed. Screening for segregation ratio distortions followed by thorough genetic analysis proved to be a powerful tool for identifying gametophytic mutations of all phases of pollen development.

scholarly journals Different Transport Mechanisms in Plant and Human AMT/Rh-type Ammonium Transporters

2006 ◽  
Vol 127 (2) ◽  
pp. 133-144 ◽  
Author(s):  
Maria Mayer ◽  
Gabriel Schaaf ◽  
Isabelle Mouro ◽  
Claude Lopez ◽  
Yves Colin ◽  
...  
Keyword(s):  
Xenopus Oocytes ◽  
Alkali Cation ◽  
Ammonium Transporter ◽  
Ammonium Transport ◽  
Transport Mechanisms ◽  
Gas Channel ◽  
Yeast Strains ◽  
Cation Conductance ◽  
Ammonium Transporters ◽  
Rh Proteins

The conserved family of AMT/Rh proteins facilitates ammonium transport across animal, plant, and microbial membranes. A bacterial homologue, AmtB, forms a channel-like structure and appears to function as an NH3 gas channel. To evaluate the function of eukaryotic homologues, the human RhCG glycoprotein and the tomato plant ammonium transporter LeAMT1;2 were expressed and compared in Xenopus oocytes and yeast. RhCG mediated the electroneutral transport of methylammonium (MeA), which saturated with Km = 3.8 mM at pHo 7.5. Uptake was strongly favored by increasing the pHo and was inhibited by ammonium. Ammonium induced rapid cytosolic alkalinization in RhCG-expressing oocytes. Additionally, RhCG expression was associated with an alkali-cation conductance, which was not significantly permeable to NH4+ and was apparently uncoupled from the ammonium transport. In contrast, expression of the homologous LeAMT1;2 induced pHo-independent MeA+ uptake and specific NH4+ and MeA+ currents that were distinct from endogenous currents. The different mechanisms of transport, including the RhCG-associated alkali-cation conductance, were verified by heterologous expression in appropriate yeast strains. Thus, homologous AMT/Rh-type proteins function in a distinct manner; while LeAMT1;2 carries specifically NH4+, or cotransports NH3/H+, RhCG mediates electroneutral NH3 transport.

scholarly journals A pore-occluding phenylalanine gate prevents ion slippage through plant ammonium transporters

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Pascal Ganz ◽  
Robin Mink ◽  
Toyosi Ijato ◽  
Romano Porras-Murillo ◽  
Uwe Ludewig ◽  
...  
Keyword(s):  
High Resolution ◽  
Protein Structures ◽  
Transport Proteins ◽  
Ammonium Ion ◽  
Ammonium Transport ◽  
Ionic Form ◽  
Potassium Ions ◽  
High Homology ◽  
Transport Capacity ◽  
Ammonium Transporters

AbstractThroughout all kingdoms of life, highly conserved transport proteins mediate the passage of ammonium across membranes. These transporters share a high homology and a common pore structure. Whether NH3, NH4+ or NH3 + H+ is the molecularly transported substrate, still remains unclear for distinct proteins. High-resolution protein structures of several ammonium transporters suggested two conserved pore domains, an external NH4+ recruitment site and a pore-occluding twin phenylalanine gate, to take over a crucial role in substrate determination and selectivity. Here, we show that while the external recruitment site seems essential for AtAMT1;2 function, single mutants of the double phenylalanine gate were not reduced in their ammonium transport capacity. Despite an unchanged ammonium transport rate, a single mutant of the inner phenylalanine showed reduced N-isotope selection that was proposed to be associated with ammonium deprotonation during transport. Even though ammonium might pass the mutant AMT pore in the ionic form, the transporter still excluded potassium ions from being transported. Our results, highlight the importance of the twin phenylalanine gate in blocking uncontrolled ammonium ion flux.

scholarly journals Abscisic-acid-dependent regulation of Arabidopsis thaliana ammonium transport relies on ABI1 control of CIPK23 and AMT1

2021 ◽  
Author(s):  
Pascal Ganz ◽  
Romano Porras-Murillo ◽  
Toyosi Ijato ◽  
Jochen Menz ◽  
Tatsiana Straub ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Abscisic Acid ◽  
Abiotic Stress ◽  
Protein Kinase ◽  
Growth Conditions ◽  
Plant Roots ◽  
Ammonium Uptake ◽  
Ammonium Transport ◽  
Ammonium Toxicity ◽  
Ammonium Transporters

SummaryAmmonium uptake at plant roots is regulated at the transcriptional, post-transcriptional and post-translational levels. Phosphorylation by the protein kinase CIPK23 transiently inactivates the ammonium transporters (AMT1s) but the phosphatases activating AMT1s remain unknown. Here, we have identified the PP2C phosphatase ABI1 as an activator of AMTs in Arabidopsis thaliana. We show that high external ammonium concentrations elevate the stress phytohormone abscisic acid (ABA) by de-glycosylation. Active ABA is sensed by ABI1-PYL complexes followed by the inactivation of ABI1 activating CIPK23. Under favourable growth conditions, ABI1 reduces AMT1 phosphorylation, both by binding and inactivating CIPK23, and by the direct dephosphorylation of AMT1s. Thus, ABI1 is a positive regulator of ammonium uptake, coupling nutrient acquisition to abiotic stress signalling. Elevated ABA reduces ammonium uptake during stress situations, such as ammonium toxicity, whereas ABI1 reactivates AMT1s under favourable growth conditions.

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