A new method for non-invasive biomass determination based on stereo photogrammetry.

A novel, non-destructive method for the biomass estimation of biological samples on culture dishes was developed. To achieve this, a photogrammetric system, which consists of a digital single-lens reflex camera (DSLR), an illuminated platform where the culture dishes are positioned and an Arduino board which controls the capturing process, was constructed. The camera was mounted on a holder which set the camera at different title angles and the platform rotated, to capture images from different directions. A software, based on stereo photogrammetry, was developed for the three-dimensional (3D) reconstruction of the samples. The proof-of-concept was demonstrated in a series of experiments with plant tissue cultures and specifically with calli cultures of Salvia fruticosa and Ocimum basilicum. For a period of 14 days images of these cultures were acquired and 3D-reconstructions and volumetric data were obtained. The volumetric data correlated well with the experimental measurements and made the calculation of the specific growth rate, µ max, possible. The µ max value for S. fruticosa samples was 0.14 day-1 and for O. basilicum 0.16 day-1. The developed method demonstrated the high potential of this photogrammetric approach in the biological sciences.

Salvia broussonetii Benth.: aroma profile and micromorphological analysis.

The volatile profiles (VOC) and the essential oil (EO) composition from the aerial parts of Salvia broussonetii were analysed. Sesquiterpene hydrocarbons dominate the VOCs from leaves (95.7%) and flowers (67.6%), followed by monoterpene hydrocarbons (2.6 and 29.7%, respectively). The main common compounds are germacrene D, ß-bourbonene, α-pinene, α-copaene and α-gurjunene, even if with divergent relative abundances. In the leaf EOs the sesquiterpenes prevail, even if not overwhelmingly (about 50.0%), followed by monoterpenes (23.0-35.0%) and by minor fractions of diterpene hydrocarbons and non-terpene derivates. The most abundant common compounds across the two sampling periods are α-pinene, ß-pinene, isobornyl acetate, α-gurjenene, germacrene D and bifloratriene. A morphological characterisation of the trichomes responsible for the productivity in terpenes was also performed. Four different morphotypes were observed on the above ground organs of S. brussonetii: peltates and capitates of type II and III resulted the only producers of volatile substances.

Monitoring of rosmarinic acid accumulation in sage cell cultures using laccase biosensor.

INTRODUCTION: A recently developed laccase based biosensor is used for polyphenols determination from in vitro Salvia cultures, the results being expressed as rosmarinic acid equivalent content. OBJECTIVE: The aim of this work was to use a previously developed laccase biosensor for the determination of total phenolic content from in vitro cultivated Salvia, and to support the biosensors further application for the assessment of polyphenols metabolites. METHODOLOGY: The biosensor was constructed by drop casting 3 µL of laccase solution and stabilisation with 0.1 % Nafion solution onto a DropSens carbon screen-printed electrode. Electrochemical measurements were carried out in a 0.1 mol/L phosphate buffer (pH 4.50), the applied working potential being -30 mV versus reference electrode. RESULTS: The response of the biosensor developed was characterised in terms of repeatability, accuracy and precision; the limit of detection was 7.5 × 10(-7) mol/L, the limit of determination was 9.5 × 10⁻7 mol/L, and linear response range for rosmarinic acid was 1 × 10⁻6-10⁻5 mol/L. CONCLUSION: A stable, sensitive and simple biosensor based on laccase-nafion was used for monitoring the total polyphenolic content from two in vitro cultivated plants. The biosensor response was free of electrochemical interferences and of possible interferences from growth media constituents, demonstrating a high sensitivity for rosmarinic acid determination in cell culture suspensions.