Electrochemical Behaviour of Tocopherols: Possibilities of Their Simultaneous Voltammetric Detection

An electroanalytical study for possible simultaneous detection of three naturally occurring isomers of vitamin E (α, γ, and δ-tocopherol) was performed. This research includes several optimization steps, such as selection of electrode material, composition of working medium, selection of electrochemical technique, and parameters of square-wave voltammetry (SWV), to reach a well-defined recognition of peaks. A glassy carbon electrode, 99.9% acetonitrile containing 0.1 mol L−1 lithium perchlorate, SWV at the potential step of 1 mV, potential amplitude of 25 mV, and frequency of 25 Hz were decided as the most suitable working conditions. Nevertheless, the corresponding anodic peaks were not sufficiently separated due to their overlapping. Thus, four standard evaluation methods (polynomial or linear baseline, zero base, and deconvolution) were compared, and the last-mentioned method was chosen as optimum. Similar linear ranges from 3.0 × 10−6 to 1.0 × 10−5 mol L−1 were obtained for α, γ, and δ-tocopherol, characterized by determination coefficient of 0.998, 0.985, and 0.994, quantification limits of 11.28, 2.70, and 3.67 × 10−6 mol L−1 and detection limits of 3.72, 0.89, and 1.21 × 10−6 mol L−1, respectively. A recovery from 72.0 to 128.5% for different concetration ratios of tocopherols has been achieved. This recovery range is in the accordance with values reported for liquid chromatography.

Whole genome sequencing of colonies derived from cannabis flowers and the impact of media selection on benchmarking total yeast and mold detection tools

Background: Cannabis products are subjected to microbial testing for human pathogenic fungi and bacteria. These testing requirements often rely on non-specific colony forming unit (CFU/g) specifications without clarity on which medium, selection or growth times are required. We performed whole genome sequencing to assess the specificity of colony forming units (CFU) derived from three different plating media: Potato Dextrose Agar (PDA), PDA with chloramphenicol and Dichloran Rose Bengal with chloramphenicol (DRBC). Methods: Colonies were isolated from each medium type and their whole genomes sequenced to identify the diversity of microbes present on each medium selection. Fungal Internal Transcribed Spacer (ITS3) and Bacterial 16S RNA(16S) quantitative polymerase chain reactions (qPCR) were performed, to correlate these CFUs with fungi- and bacterial- specific qPCR. Results: Each plating medium displayed a ten-fold difference in CFU counts. PDA with chloramphenicol showed the highest diversity and the highest concordance with whole genome sequencing. According to ITS3 and 16S qPCR confirmed with whole genome sequencing, DRBC under counted yeast and mold while PDA without chloramphenicol over counted CFUs due to bacterial growth without selection. Conclusions: Colony Forming Unit regulations lack specificity. Each medium produces significant differences in CFU counts. These are further dependent on subjective interpretation, failure to culture most microbes, and poor selection between bacteria and fungi. Given the most human pathogenic microbes found on cannabis are endophytes which culture fails to detect, molecular methods offer a solution to this long-standing quantification problem in the cannabis testing field.

Evaluation of substrate composition and exogenous hormone application on vegetative propagule rooting success of essential oil hemp (Cannabis sativa L.)

To support the rapidly expanding industrial hemp industry, a commercial supply of high-quality starter plants with low genetic variability from nurseries will be key to consistent and efficient cultivation efforts. Rooting success was evaluated across four propagation medias, five rooting hormones, and eight commercially available high-cannabidiol (CBD) essential oil hemp cultivars. Cuttings were placed in a climate-controlled room and assessed for rooting success 12 days after cloning. Rooting success was determined by quantifying total root number, cumulative total root length, and total root mass. Propagation media had the greatest effect on rooting success (13–80%). Rockwool had the highest rooting success resulting in 10-fold increases in rooting traits over the next highest scoring medium (Berger BM6). Hormone applications significantly improved (15- to 18-fold) rooting success compared to no hormone application, while non-statistical differences were observed across auxin hormone concentrations and application methods. Genetic variation in rooting response was observed between cultivars with ‘Cherry Wine’ outperforming all other cultivars with an approximate 20% increase in rooting success over the next highest rooting cultivar, ‘Wife’. Although the ideal combination was not specifically identified in this study, findings provide insight into how rooting hormone application and medium selection impact vegetative propagule rooting success of essential oil hemp.

Whole genome sequencing of colonies derived from cannabis flowers and the impact of media selection on benchmarking total yeast and mold detection tools

Background: Cannabis products are subjected to microbial testing for pathogenic fungi and bacteria. These testing requirements often rely on non-specific colony forming unit (CFU/g) specifications without clarity on which medium, selection or growth times are required. We performed whole genome sequencing to assess the specificity of colony forming units (CFU) derived from three different plating media: Potato Dextrose Agar (PDA), PDA with chloramphenicol and Dichloran Rose Bengal with chloramphenicol (DRBC). Methods: Colonies were isolated from each medium type and their whole genomes sequenced to identify the diversity of microbes present on each medium selection. Fungal Internal Transcribed Spacer (ITS3) and Bacterial 16S RNA(16S) quantitative polymerase chain reactions (qPCR) were performed, to correlate these CFUs with fungi- and bacterial- specific qPCR. Results: Each plating medium displayed a ten-fold difference in CFU counts. PDA with chloramphenicol showed the highest diversity and the highest concordance with whole genome sequencing. According to ITS3 and 16S qPCR confirmed with whole genome sequencing, DRBC under counted yeast and mold while PDA without chloramphenicol over counted CFUs due to bacterial growth without selection. Conclusions: Colony Forming Unit regulations lack specificity. Each medium produces significant differences in CFU counts. These are further dependent on subjective interpretation, failure to culture most microbes, and poor selection between bacteria and fungi. Given the most pathogenic microbes found on cannabis are endophytes which culture fails to detect, molecular methods offer a solution to this long-standing quantification problem in the cannabis testing field.

Evaluation of substrate composition and exogenous hormone application on vegetative propagule rooting success of essential oil hemp (Cannabis sativa L.)

To support the rapidly expanding industrial hemp industry, a commercial supply of high-quality starter plants with low genetic variability from nurseries will be key to consistent and efficient cultivation efforts. Rooting success was evaluated across four propagation medias, five rooting hormones, and eight commercially available high-cannabidiol (CBD) essential oil hemp cultivars. Cuttings were placed in a climate-controlled room and assessed for rooting success 12 days after cloning. Rooting success was determined by quantifying total root number, cumulative total root length, and total root mass. Propagation media had the greatest effect on rooting success (13-80 %). Rockwool had the highest rooting success resulting in 10-fold increases in rooting traits over the next highest scoring medium (Berger BM6). Hormone applications significantly improved (15- to 18-fold) rooting success compared to no hormone application, while non-statistical differences were observed across auxin hormone concentrations and application methods. Genetic variation in rooting response was observed between cultivars with ‘Cherry Wine’ outperforming all other cultivars with an approximate 20% increase in rooting success over the next highest rooting cultivar, ‘Wife’. Although the ideal combination was not specifically identified in this study, findings provide insight into how rooting hormone application and medium selection impact vegetative propagule rooting success of essential oil hemp.

Highly Efficient Biolistic Transformation of Embryogenic Cell Suspensions of Banana Via a Liquid Medium Selection System

An efficient biolistic transformation system of banana combined with a liquid medium selection system was developed during this study. An embryogenic cell suspension (ECS) of Musa acuminata cv. Baxi (AAA) was bombarded with a particle delivery system. After 7 days of restoring culture in liquid M2 medium, embryogenic cells were transferred to a liquid selection M2 medium supplemented with 10 μg/mL hygromycin for resistance screening. The untransformed cell clusters were inhibited or killed, and a small number of transformants proliferated in the liquid selection medium. After the 0th, first, second, and third generation of antibiotic screening, there were 0, 65, 212, and 320, respectively, vitality-resistant buds obtained from a 0.5-mL packed cell volume (PCV) of embryogenic cell suspension. The β-glucuronidase (GUS) staining, polymerase chain reaction (PCR) analysis, and Southern blot hybridization results all demonstrated a 100% positive rate of regenerated resistant seedlings. Interestingly, the number of buds obtained through third-generation screening was almost equal to that obtained from the original ECS in M2 medium without antibiotics. These results suggested that the liquid medium selection system facilitated the proliferation of a positive transgenic ECS, which significantly improved the regeneration rate of transformants. This protocol is suitable for the genetic transformation of all banana genotypes and is highly advantageous to varieties with low callusing potential.

Multi-Media and Multi-Band Based Adaptation Layer Techniques for Underwater Sensor Networks

In the last few decades, underwater communication systems have been widely used for the development of navy, military, business, and safety applications, etc. However, in underwater communication systems, there are several challenging issues, such as limitations in bandwidth, propagation delay, 3D topology, media access control, routing, resource utilization, and power constraints. Underwater communication systems work under severe channel conditions such as ambient noise, frequency selectivity, multi-path and Doppler shifts. In order to collect and transmit the data in effective ways, multi-media/multi-band-based adaptation layer technology is proposed in this paper. The underwater communication scenario comprises of Unmanned Underwater Vehicles (UUVs), Surface gateways, sensor nodes, etc. The transmission of data starts from sensor nodes to surface gateway in a hierarchical manner through multiple channels. In order to provide strong and reliable communication underwater, the adaptation layer uses a multi-band/multi-media approach for transferring data. Hence, in this paper, existing techniques for splitting the band such as Orthogonal Frequency-Division Multiple Access (OFDMA), Frequency-Division Multiple Access (FDMA), or Orthogonal Frequency-Division Multiplexing (OFDM) are used for splitting the frequency band, and the medium selection mechanism is proposed to carry the signal through different media such as Acoustic, Visible Light Communication (VLC), and Infrared (IR) signals in underwater. For the channel selection mechanism, two phases are involved: 1. Finding the distance of near and far nodes using Manhattan method, and 2. Medium selection and data transferring algorithm for choosing different media.