ABSTRACT
In Arabidopsis the plasma membrane nitrate transceptor (transporter/receptor) NRT1.1 governs many physiological and developmental responses to nitrate. Alongside facilitating nitrate uptake, NRT1.1 regulates the expression levels of many nitrate assimilation pathway genes, modulates root system architecture, relieves seed dormancy and protects plants from ammonium toxicity. Here, we assess the functional and phenotypic consequences of point mutations in two key residues of NRT1.1 (P492 and T101). We show that the point mutations differentially affect several of the NRT1.1-dependent responses to nitrate, namely the repression of lateral root development at low nitrate concentrations, and the short-term upregulation of the nitrate-uptake gene NRT2.1, and its longer-term downregulation, at high nitrate concentrations. We also show that these mutations have differential effects on genome-wide gene expression. Our findings indicate that NRT1.1 activates four separate signalling mechanisms, which have independent structural bases in the protein. In particular, we present evidence to suggest that the phosphorylated and non-phosphorylated forms of NRT1.1 at T101 have distinct signalling functions, and that the nitrate-dependent regulation of root development depends on the phosphorylated form. Our findings add to the evidence that NRT1.1 is able to trigger independent signalling pathways in Arabidopsis in response to different environmental conditions.
ABSTRACT
Determining the pH values of microscopic plant samples may help to explain complex processes in plants, so it is an area of interest to botanists. Fiber-optic probes with small dimensions can be used for this purpose. This paper deals with the fiber-optic detection of the pH values of droplets of plant xylem exudate based on ratiometric fluorescence intensity measurements with an internal reference. For this purpose, novel V-taper sensing probes with a minimum diameter of around 8 µm were prepared that enable the delivery of fluorescence signal from the detection site on the taper tip to the detector. The taper tips were coated with pH-sensitive transducer (8-hydroxypyrene-1,3,6-trisulfonic acid trisodium salt; HPTS) and a reference [dichlorotris-(1,10-phenanthroline) ruthenium (II) hydrate (Ru-phen dichloride)] immobilized in a xerogel layer of propyltriethoxysilane and (3-glycidoxy)propyl trimethoxysilane. The prepared probes were sensitive to pH values mainly in the range from 6.0 to 9.0. In the pH range 6-9, the results were limited by measurement errors of about 0.2 pH units, and in the pH range 5-6 by measurement errors of about 0.5 pH units. Using the developed V-taper sensing probes, the pH values of in vivo and in vitro samples of small volumes (~6 µl) of exudate were measured. The results were validated by comparison with conventional electrochemical pH measurements.
Subject(s)
Biosensing Techniques/methods , Fiber Optic Technology/methods , Cell Size , Fluorescence , Fluorescent Dyes/chemistry , Hydrogen-Ion Concentration , Xylem/chemistryABSTRACT
The PIN-FORMED (PIN) protein family is a group of plant transmembrane proteins with a predicted function as secondary transporters. PINs have been shown to play a rate-limiting role in the catalysis of efflux of the plant growth regulator auxin from cells, and their asymmetrical cellular localization determines the direction of cell-to-cell auxin flow. There is a functional redundancy of PINs and their biochemical activity is regulated at many levels. PINs constitute a flexible network underlying the directional auxin flux (polar auxin transport) which provides cells in any part of the plant body with particular positional and temporal information. Thus, the PIN network, together with downstream auxin signalling system(s), coordinates plant development. This review summarizes recent progress in the elucidation of the role of PIN proteins in polar auxin transport at the cellular level, with emphasis on their structure and evolution and regulation of their function.
