Molecular identification and physiological functional analysis of NtNRT1.1B that mediated nitrate long-distance transport and improved plant growth when overexpressed in tobacco.

Although recent physiological studies demonstrate that flue-cured tobacco preferentially utilizes nitrate ( NO 3 - ) or ammonium nitrate (NH4NO3), and possesses both high- and low-affinity uptake systems for NO 3 - , little is known about the molecular component(s) responsible for acquisition and translocation in this crop. Here we provide experimental data showing that NtNRT1.1B with a 1,785-bp coding sequence exhibited a function in mediating NO 3 - transport associated with tobacco growth on NO 3 - nutrition. Heterologous expression of NtNRT1.1B in the NO 3 - uptake-defective yeast Hp△ynt1 enabled a growth recovery of the mutant on 0.5 mM NO 3 - , suggesting a possible molecular function of NtNRT1.1B in the import of NO 3 - into cells. Transient expression of NtNRT1.1B::green fluorescent protein (GFP) in tobacco leaf cells revealed that NtNRT1.1B targeted mainly the plasma membrane, indicating the possibility of NO 3 - permeation across cell membranes via NtNRT1.1B. Furthermore, promoter activity assays using a GFP marker clearly indicated that NtNRT1.1B transcription in roots may be down-regulated by N starvation and induced by N resupply, including NO 3 - , after 3 days' N depletion. Significantly, constitutive overexpression of NtNRT1.1B could remarkably enhance tobacco growth by showing a higher accumulation of biomass and total N, NO 3 - , and even NH 4 + in plants supplied with NO 3 - ; this NtNRT1.1B-facilitated N acquisition/accumulation could be strengthened by short-term 15N- NO 3 - root influx assays, which showed 15%-20% higher NO 3 - deposition in NtNRT1.1B-overexpressors as well as a high affinity of NtNRT1.1B for NO 3 - at a K m of around 30-45 µM. Together with the detection of NtNRT1.1B promoter activity in the root stele and shoot-stem vascular tissues, and higher NO 3 - in both xylem exudate and the apoplastic washing fluid of NtNRT1.1B-transgenic lines, NtNRT1.1B could be considered as a valuable molecular breeding target aiming at improving crop N-use efficiency by manipulating the absorption and long-distance distribution/transport of nitrate, thus adding a new functional homolog as a nitrate permease to the plant NRT1 family.

Mechanism of Intermittent Deep Tillage and Different Depths Improving Crop Growth From the Perspective of Rhizosphere Soil Nutrients, Root System Architectures, Bacterial Communities, and Functional Profiles.

Long-term conventional shallow tillage reduced soil quality and limited the agriculture development. Intermittent deep tillage could effectively promote agricultural production, through optimizing soil structure, underground ecology system, and soil fertility. However, the microecological mechanism of intermittent deep tillage promoting agriculture production has never been reported, and the effect of tillage depth on crop growth has not been explored in detail. In this study, three levels of intermittent deep tillage (30, 40, and 50 cm) treatments were conducted in an experimental field site with over 10 years of conventional shallow tillage (20 cm). Our results indicated that intermittent deep tillage practices helped to improve plant physiological growth status, chlorophyll a, and resistance to diseases, and the crop yield and value of output were increased with the deeper tillage practices. Crop yield (18.59%) and value of output (37.03%) were highest in IDT-50. There were three mechanisms of intermittent deep tillage practices that improved crop growth: (1) Intermittent deep tillage practices increased soil nutrients and root system architecture traits, which improved the fertility and nutrient uptake of crop through root system. (2) Changing rhizosphere environments, especially for root length, root tips, pH, and available potassium contributed to dissimilarity of bacterial communities and enriched plant growth-promoting species. (3) Functions associated with stress tolerance, including signal transduction and biosynthesis of other secondary metabolites were increased significantly in intermittent deep tillage treatments. Moreover, IDT-30 only increased soil characters and root system architecture traits compared with CK, but deeper tillage could also change rhizosphere bacterial communities and functional profiles. Plant height and stem girth in IDT-40 and IDT-50 were higher compared with IDT-30, and infection rates of black shank and black root rot in IDT-50 were even lower in IDT-40. The study provided a comprehensive explanation into the effects of intermittent deep tillage in plant production and suggested an optimal depth.

Electrostatic Assembly of Platinum Nanoparticles along Electrospun Polymeric Nanofibers for High Performance Electrochemical Sensors.

A novel polyacrylonitrile (PAN) nanofibrous membrane conjugated with platinum nanoparticles (PtNPs) was fabricated by electrospinning and electrostatic assembly techniques. In this procedure, PAN was electrospun with 3-aminopropyltriethoxysilane (APS) together as precursor materials. First, amine groups were introduced onto PAN nanofibers, and then the as-prepared negative-charged platinum nanoparticles (PtNPs) were conjugated onto the surface of the amino-modified PAN nanofibers uniformly by the electrostatic interaction-mediated assembly. The fabricated PAN-PtNPs hybrid nanofibrous membrane was further utilized to modify the glassy carbon electrodes and was used for the fabrication of a non-enzymatic amperometric sensor to detect hydrogen peroxide (H2O2). The electrochemical results indicated that, due to the uniform dispersion of PtNPs and the electrostatic interaction between amine groups and PtNPs, the fabricated PAN-PtNPs nanofibrous membrane-based electrochemical sensor showed excellent electrocatalytic activity toward H2O2, and the chronoamperometry measurements illustrated that the fabricated sensor had a high sensitivity for detecting H2O2. It is anticipated that the strategies used in this work will not only guide the design and fabrication of functional polymeric nanofiber-based biomaterials and nanodevices, but also extend their potential applications in energy storage, cytology, and tissue engineering.

Electrospinning design of functional nanostructures for biosensor applications.

Electrospinning represents a simple and effective strategy for fabricating nanofibrous structures and materials with large surface-to-volume ratios and desirable engineered properties. Thus, incorporating nanoscale building blocks (NBBs) like nanoparticles, graphene quantum dots, carbon nanotubes, and graphene into electrospun fibers has become one of the most attention-getting research topics in the field of biosensing. However, the dispersion behavior of NBBs in the nanofibers, the limited surface area of nanofibers and the insufficient immobilization sites for tested biomolecules still restrict the better performances and broader applications of the fabricated biosensors. In this review, we present a comprehensive survey of strategies that have been utilized to fabricate functional fibrous nanostructures for the amplification of the detection signals of nanostructure-based biosensors. In particular, from the perspective of design configuration, we systematically summarized recent advances in the electrospinning fabrication of hybrid polymer nanofibers decorated with functionalized NBBs. The strategies for promoting better dispersion of NBBs in electrospun nanofibers, including direct blending before electrospinning and in situ synthesis during electrospinning, are introduced in detail. In addition, some effective processing routes for increasing immobilization sites of tested biomolecules such as arrangement of NBBs and morphological processing of nanofibers are also presented. In addition, the suitability of electrospun nanostructures for biosensors, and the advantages and disadvantages of each method for improving the biosensing performance are also discussed.

Nanoporous Carbon Nanofibers Decorated with Platinum Nanoparticles for Non-Enzymatic Electrochemical Sensing of H2O2.

We describe the preparation of nanoporous carbon nanofibers (CNFs) decorated with platinum nanoparticles (PtNPs) in this work by electrospining polyacrylonitrile (PAN) nanofibers and subsequent carbonization and binding of PtNPs. The fabricated nanoporous CNF-PtNP hybrids were further utilized to modify glass carbon electrodes and used for the non-enzymatic amperometric biosensor for the highly sensitive detection of hydrogen peroxide (H2O2). The morphologies of the fabricated nanoporous CNF-PtNP hybrids were observed by scanning electron microscopy, transmission electron microscopy, and their structure was further investigated with Brunauer-Emmett-Teller (BET) surface area analysis, X-ray photoelectron spectroscopy, X-ray diffraction, and Raman spectrum. The cyclic voltammetry experiments indicate that CNF-PtNP modified electrodes have high electrocatalytic activity toward H2O2 and the chronoamperometry measurements illustrate that the fabricated biosensor has a high sensitivity for detecting H2O2. We anticipate that the strategies utilized in this work will not only guide the further design and fabrication of functional nanofiber-based biomaterials and nanodevices, but also extend the potential applications in energy storage, cytology, and tissue engineering.