Optimized Synthesis of Layered Double Hydroxide Lactate Nanosheets and Their Biological Effects on Arabidopsis Seedlings

Background: Layered double hydroxide lactate nanosheets (LDH-lactate-NS) are powerful carriers for delivering macro-molecules into intact plant cells. In the past few years, some studies have been carried out on DNA/RNA transformation and plant disease resistance, but little attention has been paid to these factors during LDH-lactate-NS synthesis and delamination, nor has their relationship to the DNA adsorption capacity or transformation efficiency of plant cells been considered. Results: Since the temperature during delamination alters particle sizes and zeta potentials of LDH-lactate-NS products, we compared the LDH-lactate-NS stability, DNA adsorption rate and delivery efficiency of fluorescein isothiocyanate isomer I (FITC) of them, found that the LDH-lactate-NS obtained at 25℃ has the best characters for delivering biomolecules into plant cell. To understand the potential side effects and cytotoxicity of LDH-lactate-NS to plants, we compared the root growth rate between the Arabidopsis thaliana seedlings grown in the culture medium with 1-300 μg/mL LDH-lactate-NS and equivalent raw material, Mg(lactate)2 and Al (lactate)3. Phenotypic analysis showed LDH in a range of 1-300 μg/mL can enhance the root elongation, whereas the same concentration of raw materials dramatically inhibited root elongation, suggesting the nanocrystallization has a dramatical de-toxic effect to Mg(lactate)2 and Al (lactate)3. Since enhancing of root elongation by LDH is an unexpected phenomenon, we further designed experiments to investigate influence of LDH to Arabidopsis seedlings. We further used the gravitropic bending test, qRT-PCR analysis of auxin transport proteins, non-invasive micro-test technology and liquid chromatography-mass spectrometry to investigate the auxin transport and distribution in Arabidopsis root. Results indicated that LDH-lactate-NS affect root growth by increasing the polar auxin transport.Conclusions: Optimal synthesized LDH-lactate-NS can delivery biomolecules into intact plant cells with high efficiency and low cytotoxity. The working solution of LDH-lactate-NS can promote root elongation via increase the polar auxin transport in Arabidopsis roots.

Soot and charcoal are reservoirs of extracellular DNA

Soot and charcoal are carbonaceous materials widespread in the environment where they readily can come in contact with extracellular DNA shed from organisms. The adsorption at a surface protects DNA from chemical and biological degradation. However, a comprehensive insight into DNA adsorption at soot and charcoal is lacking. We measured DNA adsorption capacity at soot and charcoal as a function of solution composition, time and DNA length. We observed that the capacity for DNA is the highest at low pH, it increases with solution concentration and cation valency and that the activation energy for DNA adsorption at both soot and charcoal is ~50 kJmol-1. We demonstrate how the interaction between DNA and soot and charcoal partly occurs via terminal basepairs, suggesting that, besides electrostatic forces, hydrophobic interactions play an important role in binding. The importance of hydrophobic interactions increases as the hydrophobicity of a surface increases. Such strong binding and hydrophobic interactions need to be taken into account to improve DNA extraction protocols and for mitigation of the spread of antibiotic resistance genes in environmental matrices that contain soot and charcoal such as aerosol, wastewater and topsoil.

Clamped-end effect on static detection signals of DNA-microcantilever

Boundary constraint induced inhomogeneous effects are important for mechanical responses of nano/micro-devices. For microcantilever sensors, the clamped-end constraint induced inhomogeneous effect of static deformation, so called the clamped-end effect, has great influence on the detection signals. This paper is devoted to developing an alternative mechanical model to characterize the clamped-end effect on the static detection signals of the DNA-microcantilever. Different from the previous concentrated load models, the DNA adsorption is taken as an equivalent uniformly distributed tangential load on the substrate upper surface, which exactly satisfies the zero force boundary condition at the free-end. Thereout, a variable coefficient differential governing equation describing the non-uniform deformation of the DNA-microcantilever induced by the clamped-end constraint is established by using the principle of minimum potential energy. By reducing the order of the governing equation, the analytical solutions of the curvature distribution and static bending deflection are obtained. By comparing with the previous approximate surface stress models, the clamped-end effect on the static deflection signals is discussed, and the importance of the neutral axis shift effect is also illustrated for the asymmetric laminated microcantilever.

Sequence-Independent DNA Adsorption on Few-Layered Oxygen-Functionalized Graphene Electrodes: An Electrochemical Study for Biosensing Application

DNA is strongly adsorbed on oxidized graphene surfaces in the presence of divalent cations. Here, we studied the effect of DNA adsorption on electrochemical charge transfer at few-layered, oxygen-functionalized graphene (GOx) electrodes. DNA adsorption on the inkjet-printed GOx electrodes caused amplified current response from ferro/ferricyanide redox probe at concentration range 1 aM–10 nM in differential pulse voltammetry. We studied a number of variables that may affect the current response of the interface: sequence type, conformation, concentration, length, and ionic strength. Later, we showed a proof-of-concept DNA biosensing application, which is free from chemical immobilization of the probe and sensitive at attomolar concentration regime. We propose that GOx electrodes promise a low-cost solution to fabricate a highly sensitive platform for label-free and chemisorption-free DNA biosensing.

Influence of the Electrolyte Salt Concentration on DNA Detection with Graphene Transistors

Liquid-gated Graphene Field-Effect Transistors (GFET) are ultrasensitive bio-detection platforms carrying out the graphene’s exceptional intrinsic functionalities. Buffer and dilution factor are prevalent strategies towards the optimum performance of the GFETs. However, beyond the Debye length (λD), the role of the graphene-electrolytes’ ionic species interactions on the DNA behavior at the nanoscale interface is complicated. We studied the characteristics of the GFETs under different ionic strength, pH, and electrolyte type, e.g., phosphate buffer (PB), and phosphate buffer saline (PBS), in an automatic portable built-in system. The electrostatic gating and charge transfer phenomena were inferred from the field-effect measurements of the Dirac point position in single-layer graphene (SLG) transistors transfer curves. Results denote that λD is not the main factor governing the effective nanoscale screening environment. We observed that the longer λD was not the determining characteristic for sensitivity increment and limit of detection (LoD) as demonstrated by different types and ionic strengths of measuring buffers. In the DNA hybridization study, our findings show the role of the additional salts present in PBS, as compared to PB, in increasing graphene electron mobility, electrostatic shielding, intermolecular forces and DNA adsorption kinetics leading to an improved sensitivity.