Sensitivity and specificity improvement of an ion sensitive field effect transistors-based biosensor for potato glycoalkaloids detection

Butyryl cholinesterase of different origin along with variations of the time of enzyme immobilization on the potentiometric transducer surface is offered to control the ion sensitive field effect transistor (ISFET)-based biosensor sensitivity. Because butyryl cholinesterase has been already used to develop the sensors for heavy metals, organophosphorus/carbamate pesticides, and steroidal glycoalkaloids analysis, the present study has been focused on the investigation and adjustment of the ISFET-based biosensor specificity exclusively to the glycoalkaloids. Utilization of ethylendiaminetetracetate (a complexon of heavy metal ions) and phosphotriesterase (a highly efficient catalyst for the hydrolysis of organophosphorus compounds) enabled the highly specific determination of glycoalkaloids at the background of lead and mercury (up to 500 microM of ions concentration) and paraoxon (up to 100 microM of pesticide concentration). The developed biosensor has been validated for glycoalkaloids detection in potato varieties cultivated in Ukraine, and the results obtained are compared to those measured by the methods of HPLC and TLC.     

Citrus genebank collections: international collaboration opportunities between the US and Russia

Citrus germplasm is conserved in genebanks at sites around the world to provide genetic resources for breeding and research programs. The value of genebank collections is particularly evident as diseases and climate change threaten citrus production areas. We provide historical, inventory, and maintenance information about national citrus collections in Russia and in the United States. The Russian Research Institute of Floriculture and Subtropical Crops (RRIFSC) in Sochi, Russia maintains a collection of 132 citrus accessions representing 50 taxa. Southern Russia is one of the most northerly citrus growing areas in the world and many accessions in the RRIFSC collection were specifically selected for cold tolerance to facilitate citrus production in this region. Tree response data taken during the frequent severe winters in Sochi has provided an understanding of the relative cold tolerance of many RRIFSC accessions. Particularly noteworthy is the array of cold-tolerant lemon accessions maintained at the RRIFSC. The Aurantioideae collections at the United States Department of Agriculture (USDA), Agricultural Research Service (ARS) National Clonal Germplasm Repository for Citrus and Dates (NCGRCD) and at the University of California Citrus Variety Collection in Riverside, California, maintain 1328 accessions of citrus cultivars and wild relatives. Because of federal and state quarantine regulations, accessions at this facility are tested for graft-transmissible pathogens and undergo therapy to eliminate known pathogens. In contrast to Sochi, desert-adapted lemon and grapefruit varieties are a major interest of the California-based collection. The collection of citrus and citrus relatives is being screened for resistance and/or tolerance genes to diseases such as huanglongbing which is currently threatening the US citrus industry. Through collaborative exchange efforts, the diversity in the two genebanks could be expanded to increase availability of desirable genetic resources to breeding and research communities throughout the world.     

Kinetics of human and horse sera cholinesterases inhibition with solanaceous glycoalkaloids: Study by potentiometric biosensor

The kinetics of inhibition of human and horse sera butyryl cholinesterases by solanaceous glycoalkaloids α-solanine, α-chaconine and tomatine has been studied by means of a potentiometric biosensor based on pH-sensitive field effect transistors (pH-FETs). Using acetyl- and butyryl choline as substrates, the optimal pH and the apparent kinetic parameters (K_mapp, V_maxapp) of immobilized cholinesterases have been calculated in the absence of inhibitors. All studied glycoalkaloids were reversible inhibitors of both butyryl cholinesterases, and inhibited the horse and human immobilized enzymes in competitive and mixed modes, respectively. The affinity of each enzyme towards α-solanine, α-chaconine and tomatine has been estimated through calculation of apparent inhibition constants K_iapp and inhibition coefficients I₅₀. An application of the butyryl cholinesterases studied in the biosensors for glycoalkaloids determination in the concentration range of 10⁻⁷ to 10⁻⁴ M has been discussed.