μM Silver Research Articles

Improved Protocol for Efficient Agrobacterium-Mediated Transient Gene Expression in Medicago sativa L.

Medicago sativa L. (Alfalfa) is a globally recognized forage legume that has recently gained attention for its high protein content, making it suitable for both human and animal consumption. However, due to its perennial nature and autotetraploid genetics, conventional plant breeding requires a longer timeframe compared to other crops. Therefore, genetic engineering offers a faster route for trait modification and improvement. Here, we describe a protocol for achieving efficient transient gene expression in alfalfa through genetic transformation with the Agrobacterium tumefaciens pCAMBIA1304 vector. This vector contains the reporter genes β-glucuronidase (GUS) and green fluorescent protein (GFP), along with a selectable hygromycin B phosphotransferase gene, all driven by the CaMV 35s promoter. Various transformation parameters-such as different explant types, leaf ages, leaf sizes, wounding types, bacterial concentrations (OD600nm), tissue preculture periods, infection periods, co-cultivation periods, and different concentrations of acetosyringone, silver nitrate, and calcium chloride-were optimized using 3-week-old in vitro-grown plantlets. Results were attained from data based on the semi-quantitative observation of the percentage and number of GUS spots on different days of agro-infection in alfalfa explants. The highest percentage of GUS positivity (76.2%) was observed in 3-week-old, scalpel-wounded, segmented alfalfa leaf explants after 3 days of agro-infection at a bacterial concentration of 0.6, with 2 days of preculture, 30 min of co-cultivation, and the addition of 150 µM acetosyringone, 4 mM calcium chloride, and 75 µM silver nitrate. The transient expression of genes of interest was confirmed via histochemical GUS and GFP assays. The results based on transient reporter gene expression suggest that various factors influence T-DNA delivery in the Agrobacterium-mediated transformation of alfalfa. The improved protocol can be used in stable transformation techniques for alfalfa.

Effect of Packaging Material and Storage Period on Popping Behaviour of Roasted Makhana Seeds

The effect of storage period and type of packaging material on the popping behaviour of roasted makhana seeds was studied. Raw makhana seeds with a diameter of 9-10 mm were roasted using a batch-type primary roasting machine and tempered for 48 hours to bring the roasted seed moisture content to about 16% dry basis (d.b.). The roasted seeds were stored in 15, 25, and 80 µm Low-Density Polyethylene (LDPE), 110 µm Polypropylene (PP), 140 µm Biaxially Oriented Polypropylene (BOPP), and 60 μm Silver Laminate (SL) packets for six months. Popping of the stored seeds was conducted at 15-day intervals, and observations were made on popped makhana recovery, popping efficiency, colour of popped makhana, and expansion characteristics. The expansion ratio, expansion index, popped makhana recovery and popping efficiencies prior to storage were 2.37±0.22, 13.64±4.04, 48.53±0.50, and 82.89±0.78%, respectively. Duncan’s multiple range tests showed that the seeds packed in 15 µm LDPE packages maintained the quality and recovery up to 60 days only. The 80 μm LDPE packets, BOPP, PP, and SL packets were found effective in maintaining popping efficiency, recovery of popped makhana, and expansion ratio more than 81.44±0.70%, 47.95±0.43%, and 2.14±0.14, respectively with no significant colour change up to 135 days storage of the roasted seeds. Significant changes in colour values of popped makhana were observed after 135 days of storage and a* value increased whereas L* and b* values decreased. This study showed that makhana seeds may be popped effectively with >47% popped makhana recovery, >16 mm average diameter and acceptable colour values up to 120 days when the roasted seeds are stored at 15.53±0.49% (d.b.) in airtight packaging material, which was not possible until now.

Screening the Performance of a Reverse Osmosis Pilot-Scale Process That Treats Blended Feedwater Containing a Nanofiltration Concentrate and Brackish Groundwater.

A two-stage pilot plant study has been completed that evaluated the performance of a reverse osmosis (RO) membrane process for the treatment of feedwater that consisted of a blend of a nanofiltration (NF) concentrate and brackish groundwater. Membrane performance was assessed by monitoring the process operation, collecting water quality data, and documenting the blended feedwater's impact on fouling due to microbiological or organic means, plugging, and scaling, or their combination. Fluorescence and biological activity reaction tests were used to identify the types of organics and microorganisms present in the blended feedwater. Additionally, scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS) were used to analyze suspended matter that collected on the surfaces of cartridge filters used in the pilot's pretreatment system. SEM and EDS were also used to evaluate solids collected on the surfaces of 0.45 µm silver filter pads after filtering known volumes of NF concentrate and RO feedwater blends. Water quality analyses confirmed that the blended feedwater contained little to no dissolved oxygen, and a significant amount of particulate matter was absent from the blended feedwater as defined by silt density index and turbidity measurements. However, water quality results suggested that the presence of sulfate, sulfide, iron, anaerobic bacteria, and humic acid organics likely contributed to the formation of pyrite observed on some of the membrane surfaces autopsied at the conclusion of pilot operations. It was determined that first-stage membrane productivity was impacted by the location of cartridge filter pretreatment; however, second-stage productivity was maintained with no observed flux decline during the entire pilot operation's timeline. Study results indicated that the operation of an RO process treating a blend of an NF concentrate and brackish groundwater could maintain a sustainable and productive operation that provided a practical minimum liquid discharge process operation for the NF concentrate, while the dilution of RO feedwater salinity would lower overall production costs.

Emission of ions and electrons correlated with soft and hard x-rays evolution from thermal plasma

This paper presents a correlation between time evolution of ions and electrons with soft and hard x-rays emitted from argon plasma. The plasma setup comprises of two copper electrodes connected with a dc power supply. Faraday cups were used to monitor time evolution of ions, to extract their energy, temperature, and flux. Double Langmuir probe was employed to determine electron temperature, energy, and density. To explore time-resolved emission of soft x-rays, PIN photodiodes filtered with 24 μm aluminum, 90 μm Mylar, 80 μm copper, and 10 μm silver have been used. To evaluate temporal evolution of hard x-rays, a scintillator–photomultiplier system was utilized. The plasma was generated using argon gas at atmospheric pressure 760 Torr and constant flow rate 5 L/min at 7, 9, and 11 kV. The flux, energy, and temperature of Ar ions increase with the increase in the operating potential. Soft x-ray signals last much longer (3000 ns) than those of electrons' and ions' signals (300 ns). The plasma operated at 11 kV permits highest emission of ions, electrons, and x-rays. The ions, electrons, and hard and soft x-ray irradiation on silicon wafer exhibited the presence of damaged trails. Ion irradiation showed the presence of latent damaged trails. Electron irradiation caused more damage to the Si surface compared to ion irradiation due to higher flux and density. Soft x-rays had a lesser effect as compared to Si exposed to hard x-rays due to higher energy of the hard x-rays. In the case of hard x-rays irradiated Si, erupted volcano-like structure is formed.

Evaluation of Parameters Affecting Agrobacterium-Mediated Transient Gene Expression in Industrial Hemp (Cannabis sativa L.).

Industrial hemp Cannabis sativa L. is an economically important crop mostly grown for its fiber, oil, and seeds. Due to its increasing applications in the pharmaceutical industry and a lack of knowledge of gene functions in cannabinoid biosynthesis pathways, developing an efficient transformation platform for the genetic engineering of industrial hemp has become necessary to enable functional genomic and industrial application studies. A critical step in the development of Agrobacterium tumefaciens-mediated transformation in the hemp genus is the establishment of optimal conditions for T-DNA gene delivery into different explants from which whole plantlets can be regenerated. As a first step in the development of a successful Agrobacterium tumefaciens-mediated transformation method for hemp gene editing, the factors influencing the successful T-DNA integration and expression (as measured by transient β-glucuronidase (GUS) and Green Florescent Protein (GFP) expression) were investigated. In this study, the parameters for an agroinfiltration system in hemp, which applies to the stable transformation method, were optimized. In the present study, we tested different explants, such as 1- to 3-week-old leaves, cotyledons, hypocotyls, root segments, nodal parts, and 2- to 3-week-old leaf-derived calli. We observed that the 3-week-old leaves were the best explant for transient gene expression. Fully expanded 2- to 3-week-old leaf explants, in combination with 30 min of immersion time, 60 µM silver nitrate, 0.5 µM calcium chloride, 150 µM natural phenolic compound acetosyringone, and a bacterial density of OD600nm = 0.4 resulted in the highest GUS and GFP expression. The improved method of genetic transformation established in the present study will be useful for the introduction of foreign genes of interest, using the latest technologies such as genome editing, and studying gene functions that regulate secondary metabolites in hemp.

Silver-TiO2 coated D-shaped photonic crystal fiber based SPR sensor for ultrasensitive refractive index detection: design and FEM analysis

Photonic crystal fiber (PCF) based surface plasmon resonance (SPR) sensors are emerging as a promising technology for ultrasensitive detection of various biological and chemical analytes. This paper presents a novel D-shaped PCF based SPR sensor, which has shown great potential for highly sensitive detection of refractive index (RI) changes. The D-shaped configuration is achieved through the polishing of the upper side of the PCF fiber. To enhance sensitivity, a 0.1 μm silver layer is strategically placed between the fiber and analyte, intensifying light–matter interactions. Additionally, a 0.05 μm titanium dioxide (TiO2) layer is employed not only to further boost sensitivity but also to shield the metal from oxidation, ensuring the longevity and stability of the sensor. The finite element method (FEM) is employed to optimize the structural parameters of the sensor design. The findings demonstrate that the proposed SPR sensor is sensitive to RI changes in the 1.31–1.35 range, achieving a peak wavelength sensitivity of 30000 nm RIU−1 and an amplitude sensitivity of −185.33 RIU−1. The sensor holds promise for diverse applications, including chemical and biological sensing, making it a versatile tool with promising implications for advancing sensing technologies in various domains.

Development and Performance Evaluation of Stretchable Silver Pastes for Screen Printing on Thermoplastic Polyurethane Films

Efficient, stretchable wiring electrodes are achieved when the resistance change during expansion and contraction is minimal. Herein, we prepared silver pastes specifically designed for screen printing on thermoplastic polyurethane films; they exhibit minimal resistance changes. The pastes were prepared using silver particles with sizes of 2 and 7 μm as well as a mixture of 2 and 7 μm silver particles (50:50 wt%). These pastes were analyzed using methods such as rheological measurements, thermogravimetric analysis, printability tests, tensile and torsion tests, and light-emitting diode (LED) tests. The most promising results were obtained when exclusively using 2 μm silver flake particles. The pastes demonstrated a viscosity of 24,880 cps, a thixotropic index value of 2.82, excellent printability, and consistent resistance measurements even after 100% stretch, thus indicating exceptional tensile properties. Moreover, the pastes exhibited substantial stability, with no change in brightness after the attachment of seven LEDs at 20% tension.

Screening of Cork Oak for Resistance to Phytophthora cinnamomi and Micropropagation of Tolerant Seedlings

Massive propagation of cork oak (Quercus suber) individuals tolerant to Phytophthora cinnamomi (Pc) is probably the most important challenge for cork production. Screening for resistance to Pc of ca. 200 seedlings obtained from a single cork oak tree that has survived the epidemic was performed by soil infestation. Twenty months after Pc inoculation, 33 seedlings survived from Pc infection and the four most vigorous seedlings were selected. The plants were forced to produce new shoots under controlled climatic conditions, and the new shoots were used to establish the plants in vitro by axillary budding. High axillary shoot proliferation rates were achieved by culturing the new shoots on Lloyd and McCown (WPM) medium, followed by subculturing for 2 weeks on 0.22 µM benzyladenine (BA) and for 2 weeks further on 0.04 µM BA. Addition of 20 µM silver thiosulphate (STS) increased the proliferation rates and improved the appearance and development of shoots. Rooting rates of 80–100% were obtained by culturing the shoots for 24 or 48 h on Gresshoff and Doy medium with ⅓ macronutrients plus 122.5 µM indole-3-butyric acid and subsequent transfer to root expression medium containing 20 µM STS. The results of this study optimize the micropropagation of a relevant and recalcitrant tree species in forestry.

Development of 3D-Printed Collagen Scaffolds with In-Situ Synthesis of Silver Nanoparticles.

UV-irradiation method has grown as an alternative approach to in situ synthetize silver nanoparticles (AgNPs) for avoiding the use of toxic reducing agents. In this work, an antimicrobial material by in situ synthesizing AgNPs within 3D-printed collagen-based scaffolds (Col-Ag) was developed. By modifying the concentration of AgNO3 (0.05 and 0.1 M) and UV irradiation time (2 h, 4 h, and 6 h), the morphology and size of the in situ prepared AgNPs could be controlled. As a result, star-like silver particles of around 23 ± 4 μm and spherical AgNPs of 220 ± 42 nm were obtained for Ag 0.05 M, while for Ag 0.1 M cubic particles from 0.3 to 1.0 μm and round silver precipitates of 3.0 ± 0.4 μm were formed in the surface of the scaffolds at different UV irradiation times. However, inside the material AgNPs of 10-28 nm were obtained. The DSC thermal analysis showed that a higher concentration of Ag stabilizes the 3D-printed collagen-based scaffolds, while a longer UV irradiation interval produces a decrease in the denaturation temperature of collagen. The enzymatic degradation assay also revealed that the in situ formed AgNPs act as stabilizing and reinforcement agent which also improve the swelling capacity of collagen-based material. Finally, antimicrobial activity of Col-Ag was studied, showing high bactericidal efficiency against Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) bacteria. These results showed that the UV irradiation method was really attractive to modulate the size and shape of in situ synthesized AgNPs to develop antimicrobial 3D-printed collagen scaffolds with different thermal, swelling and degradation properties.

Analysis and study of the problems on the wires used in the MEG CDCH and the construction of the new drift chamber

In the MEG II detector, the measurement of the momentum of the charged particle is performed by a high transparency single volume, full stereo cylindrical Drift Chamber (CDCH). It is composed by 9 concentric layers, each consisting of 192 drift cells. The single drift cell is approximately squared, with a 20 μm gold plate tungsten sense wire surrounded by 40 μm/50 μm silver plated aluminum field wires in a ratio of 5:1. During the construction of the first CDCH, we observed the breaking of about hundred cathode wires: 97 of these were 40 μm aluminum wires, while 10 were 50 μm wires. Since the number of broken cathodes is less than 1% of the total, one can expect the influence on the track reconstruction efficiency to be not so dramatic. We verified by means of simulations that the loss of one cathode does not change the cell electric field appreciably. Here we present the results of the analysis of the effects of mechanical stress and chemical corrosion observed on these broken wires. Finally, we show the studies carried out on new wires to overcome the weaknesses found and the process that will be used for the construction of the new drift chamber (CDCH2). It will be built with the same modular technique, as for the previous one, the use of the wiring robot will be optimized to improve some weaker step in the procedure, new wires will be adopted with a 25% thicker diameter, which has very little effects on the resolution and efficiency of the detector. Furthermore these wires are made with a manufacturing process different from that used previously.

Improving culture initiation of mature oak shoots through use of silver thiosulfate.

PremiseOf the approximately 430 species of oaks (Quercus spp.) that have been assessed, 31% are threatened with extinction and in need of safeguarding. However, oak seeds cannot be seed banked, and thus rely on alternative strategies such as in vitro culture for ex situ conservation. One challenge to this approach is low culture initiation rates. Our objective was to identify factors that may improve the establishment of shoot cultures in vitro using new growth collected from mature trees.MethodsShoot cuttings were harvested from individuals of five different oak species (Q. alba, Q. bicolor, Q. macrocarpa, Q. muehlenbergii, and Q. palustris). Shoots were cultured onto medium with or without 50 µM silver thiosulfate (STS), a known inhibitor of the stress hormone ethylene. Cultures were grown for one month, at which point shoots were assessed for survival.ResultsShoot survival was significantly greater in shoots cultured on medium containing STS compared to the control group, with the overall survival rate increasing from 65% to 73%.DiscussionIncreasing the survival rate of newly established cultures is important in ensuring that material collected from endangered species has the best chance for survival, which is critical for successful ex situ conservation.

Numerical Simulation, Analysis, and Fabrication of MEMS-Based Solid Ag and Cu Microneedles for Biomedical Applications

Microelectromechanical system (MEMS)-based devices have gained attention recently due to their beneficial biomedical applications. MEMS-based devices like microneedles have set new trends in drug delivery, vaccination, skin, and eye treatment. Different materials like metals, sugars, polymers, and silicon have been used for fabrication. Various techniques have been used for their fabrication, including laser ablation, lithography, injection molding, and additive manufacturing. The tip diameter of different micron ranges has been achieved. The strength and stiffness of the microneedle’s tip have always been important in fabricating microneedles so that it does not break on insertion. This research paper presents a comparison between silver (Ag) and copper (Cu) solid microneedles by performing numerical analysis using the fuzzy approach, structural simulation, and fabrication. Firstly, structural simulation has been performed in ANSYS software to test the strength of silver (Ag) and copper (Cu) microneedles separately. The purpose is to compare the stress effect and fracture limit of both microneedles. The results collected from the simulation provide valuable target and prediction facts to fabricate improved designs of the solid Ag and Cu microneedles. Then, fuzzy-based numerical analysis has been performed in MATLAB software for both microneedles separately. In this numerical analysis, the effect on the range of microneedle tip diameter and cone length has been observed by varying input voltage and time. Finally, fabrication has been performed using a novel economical technique such as electrochemical etching. Electrochemical etching is a very low-cost and clean room-free technique as compared to other techniques used for the fabrication of microneedles. The fabrication technique adopted in this work is the same for both silver and copper microneedles. The scanning electron microscopy (SEM) characterization has been performed for both fabricated microneedle tips. The tip of the fabricated solid Ag and Cu microneedle has been then coated with drugs using the dip-coating method. The coated solid Ag and Cu microneedle’s tip has been then characterized again using SEM. The numerical results calculated from the fuzzy analysis have been then compared with fabrication results. The fuzzy analysis gives the simulated size of the microneedle’s tip for 5.05 μm silver and 5.12 μm copper which have very close approximation with the experimental values from the SEM micrographs which also give the values of the cone length from 400 to 500 μm and the tip size from 5 to 6 μm for the time of 10–15 minutes, whose values were optimized by the fuzzy analysis. The results of this research provide valuable benchmark and prediction data to fabricate improved designs of the silver solid microneedles for drug delivery and other biomedical applications.

M-brigde- and elicitor-assisted enhanced post-storage germination of Rauvolfia serpentina synthetic seeds, their genetic fidelity assessment and reserpine estimation

The present study reports an amelioration of synthetic seed germination efficiency with a simple intervention of filter paper (M-bridge) drenched in ½Murashige and Skoog liquid (½MSL) medium in Rauvolfia serpentina. Synthetic seeds were developed from in vitro shoot tips using 3% (w/v) sodium alginate and 75 mM calcium chloride. Media for synthetic seed gemination were tested in cotton bed drenched with sterile water (SW), cotton bed drenched with ½MSL medium, M-bridge drenched with ½MSL medium, and ½MS semisolid (½MSS) medium alone. Synthetic seeds inoculated on ½MSL or SW or ½MSS media, exhibited slower germination and inferior performance; however, with the involvement of M-bridge, fastest germination (5.6 ± 0.4 days) and maximum proliferation (4.8 ± 0.7 shoots with 42.8 ± 2.4 mm length, and 19.8 ± 2.7 leaves/synthetic seed after 42 days of inoculation) were recorded. Among the tested elicitors (silver nitrate, salicylic acid and sodium nitroprusside) at their different concentrations supplemented in ½MSL medium (with M-bridge), 50 µM silver nitrate resulted in 100% germination frequency and resulted in germination within 5 days. Further, the synthetic seeds were stored for 42 days at − 20 °C, 8 °C, and 25 °C. Maximum germination efficiency was obtained in case of seeds stored at 8 °C with respect to earliness in germination (6.9 ± 0.3 days), germination percentage (71.8 ± 3.4%) and subsequent proliferation (4.9 ± 0.6 shoots with 18.9 ± 0.9 mm length, and 8.9 ± 0.7 leaves/synthetic seed). Clonal fidelity analysis of the in vitro regenerants performed using RAPD markers exhibited monomorphic banding pattern. The reserpine content of the regenerants was found to be maximum following 42 days-storage at 8 °C (126.9 µg/g), followed by a substantial decline in the synthetic seed-derived regenerants following 42-days-storage at 25 °C (120.7 µg/g) or − 20 °C (121.8 µg/g).

Effect of amino acid supplementation, elicitation and LEDs on Hylocereus costaricensis callus culture for the enhancement of betalain pigments

Hylocereus costaricensis more popularly known as the purplish-red pitaya or dragon fruit belongs to the Cactaceae family. The present study analyzes the effect of amino acids, elicitors, and light quality to improve callus proliferation, enhance betalain content and antioxidant profiles through elicitation and amino acid supplementation. The treatments that stimulated the highest total betalain content in red callus cultures were 50 µM tyrosine that produced a 1.6-fold increase (11.28 ± 0.38 mg/g DM) in betalain content and 200 µM leucine that produced 2.56-fold increase (18.13 ± 5.98 mg/g DM) in betalain content. Elicitation of yellow callus cultures with 100 mg/L yeast extract enhanced betalain content by 2-fold without affecting callus biomass. Betalain content could also be improved by 1.23-fold with 20 µM silver nitrate (12.51 ± 0.55 mg/g DM) while callus dry mass and total phenolic content were elevated at 40 µM silver nitrate. The utilization of light was a promising strategy for increased production of secondary metabolites in callus cultures of H. costaricensis. Both yellow and red callus cultures incubated under red LED lights enhanced total betalain content by 3.8- fold (11.64 ± 0.54 mg/g DM) and 4.8-fold (9.42 ± 1.48 mg/g DM), significantly higher (p < 0.01) than cool white LED lights (control treatment). Amino acid supplementation and elicitation strategies in this study achieved betalain content in yellow and red callus that was higher than that in dragon fruit peel (6.31 ± 1.82 mg/g DM), the pulp (3.26 ± 0.26 mg/g DM), and in vitro seedlings (2.01 ± 0.12 mg/g DM). The present investigation reports a simple and efficient protocol for in vitro betalain production from H. costaricensis. This study may enable the future commercial production of betalain compounds under a controlled environment, independent of climate and soil conditions.

Effective reversal of hyperhydricity leading to efficient micropropagation of Dianthus chinensis L.

The online version contains supplementary material available at 10.1007/s13205-021-02645-7.

Assessment of metallic nanoparticles as radioenhancers in gastric cancer therapy by Geant4 simulation and local effect model

Gastric cancer ranks as one of the most common and poor prognosis human malignancies. We investigated the radioenhancer effects of gold, silver, platinum, bismuth, iron oxide, titanium oxide and germanium nanoparticles on simulated gastric adenocarcinoma cells, irradiated with 6 MV X-rays in presence of 0.05 µM, 0.25 µM and 0.5 µM homogeneous distributed metal nanoparticles throughout the cell, including the nucleus, cytoplasm and extracellular matrix using Geant4 Monte Carlo simulation along with LEM radiobiological model. The survival reduction coefficient was obtained 0.5 for light nanoparticles including 0.5 μM titanium oxide, iron oxide and germanium and also 0.3–0.1 for heavy nanoparticles including 0.5 μM gold, silver, bismuth and platinum. Nanoparticle-mediated Enhancement Ratio for 0.5 μM internuclear Gold, Silver, Bismuth, Germanium, Platinum, Iron oxide and Titanium oxide nanoparticles was calculated to 0.10, 0.27, 0.26, 0.47, 0.09, 0.50, and 0.58, respectively. Radiosensitization by NPs may enhance therapeutic efficacy of gastric adenocarcinoma.

Contactless electroreflectance spectroscopy with a semitransparent capacitor made of a silver mesh of ultrathin lines

Semitransparent electrodes operating in the broad spectral region are needed for many applications and one of them is contactless electroreflectance (CER) spectroscopy. In this work, the technology of printed electronics has been applied to fabricate mesh-like semitransparent electrodes composed of ultrathin (10 µm) silver lines of different densities. These electrodes have been printed on borosilicate glass and used in CER measurements. It has been shown that the CER signal is linearly proportional to the applied voltage and inversely proportional to the distance. In addition, a shadow effect has been observed for some distances because of non-transparent silver paths. Finally, a phenomenological formula has been proposed to describe the intensity of the CER signal in the capacitor with the semitransparent electrode. Moreover, the mesh-like printed electrodes have been applied to measure CER spectra for van der Waals crystals and perspectives for further development of semitransparent electrodes for CER measurements have been discussed.

Vegetative Propagation of Phytophthora cinnamomi-Tolerant Holm Oak Genotypes by Axillary Budding and Somatic Embryogenesis

Holm oak (Quercus ilex) is one of the most widely distributed tree species in the Mediterranean basin. High mortality rates have been observed in holm oak populations in the southwest of the Iberian Peninsula as a result of oak decline syndrome. Selection and propagation of genotypes tolerant to this syndrome could aid the restoration of affected areas. In this article, we report micropropagation and conservation procedures based on axillary budding and somatic embryogenesis (SE) of holm oak plants, selected for their tolerance to Phytophthora cinnamomi—the main biotic factor responsible for oak decline. Forced shoots were obtained from potted plants of eight different genotypes, and used as stock material to establish in vitro shoot proliferation cultures. Reliable shoot proliferation was obtained in seven out the eight genotypes established in vitro, whereas multiplication rates were genotype-dependent. The highest rooting rates were obtained by culturing shoots for 24 h or 48 h on rooting induction medium containing 25 mg L−1 indole-3-butyric acid, followed by transfer to medium supplemented with 20 µM silver thiosulphate. Axillary shoot cultures can be successful conserved by cold storage for 12 months at 4 °C under dim lighting. Shoot tips, excised from axillary shoot cultures established from tolerant plants, were used as initial explants to induce SE. Somatic embryos and/or nodular embryogenic structures were obtained on induction medium with or without indole-acetic acid 4 mg L−1, in two out the three genotypes evaluated, and induction rates ranged between 2 and 4%. Plantlet recovery was 45% after two months cold stratification of somatic embryos and eight weeks of culture on germination medium. Vegetative propagation of P. cinnamomi-tolerant Q. ilex trees is a valuable milestone towards the restoration of disease-affected areas.

Silver compounds regulate leaf drop and improve in vitro regeneration from mature tissues of Australian finger lime (Citrus australasica)

Finger lime is a small-leaved citrus native to Australia that is becoming increasingly popular worldwide. Finger lime is monoembryonic and cannot be propagated true to type from seeds. Initial studies to develop a tissue culture protocol for the mass production of elite lines being developed in our breeding program were unsuccessful due to complete leaf abscission during the culture establishment phase. In this study, silver-containing compounds such as silver thiosulfate (STS), silver nitrate (AgNO3), and silver nanoparticles (AgNPs) were applied at three concentrations (20, 40, and 60 µM) to manage ethylene biosynthesis in finger lime tissue cultures. The Murashige and Skoog (MS) medium supplemented with 2.2 µM BAP and 60 µM silver thiosulfate was the best medium for culture establishment. The same level of STS with 0.70 µM GA3 produced the largest number of shoots and prevented leaf abscission. Micropropagated shoots were successfully rooted in half-strength MS medium supplemented with 0.10 µM NAA. The addition of 60 µM silver thiosulfate (STS) to the MS medium resulted in efficient micropropagation of the finger lime explants and controlled leaf abscission. The study evaluates the efficacy of various silver-containing compounds on their ability to effectively control ethylene production and allow for the development of an efficient tissue culture protocol for Finger lime.

Silver nanorods induced oxidative stress and chromosomal aberrations in the Allium cepa model.

The production of different size and shape silver nanoparticles (AgNPs) has increased considerably in recent years due to several commercial and biological applications. Here, rod-shaped AgNPs (SNRs) were prepared using the microwave-assisted method and characterised by ultraviolet-visible spectroscopy, and transmission electron microscopy analysis. The present study aims to investigate the cyto-genotoxic effect of various concentrations (5, 10, and 15 µM) of SNRs using Allium cepa model. As a result, concentration-dependent cyto-genotoxic effect of SNRs was observed through a decrease in the mitotic index, and an increase in the chromosomal aberrations such as chromosome break, disturbed metaphase, and anaphase bridge. To check the impact of Ag+ ions, 15 µM silver nitrate (AgNO3) was prepared and tested in all the assays. Furthermore, cell viability and different reactive oxygen species assays were performed to test the cytotoxicity evaluation of SNRs. The authors found that in all the tested assays, SNRs at high concentrations (15 µM) and AgNO3 (15 µM) were observed to cause maximal damage to the roots. Therefore, the current study implies that the cytotoxicity and genotoxicity of SNRs were dependent on the concentration of SNRs.