METHOD OF MEASUREMENT OF NITRATE REDUCTASE ACTIVITY IN FIELD CONDITIONS

For the last three decades the interest in biomonitoring and ecological studies has been rapidly growing. Therefore, it was necessary develop of new methods of analysis for biochemical parameters which allow to quantify biological response of investigated organisms for environmental factors. The main goal of this paper demonstrates opti-mal conditions for enzyme kinetics analysis conducted in the fieldin situ.Nitrate reductase activity is typically assayed in vivo by measuring nitrite production in tissue which has been vacuum infiltratedwith buffered nitrate solution. For this study a nitrate reductase assay was adapted from a number of studies with own modi-ficationsof authors. Leaves of examined plants were collected from the investigated plots and immediately placed into test tubes with buffer solution (potassium phos-phate dibasic containing 0.6% propanol-1) and evacuated in 0.33 atm. for 10 minutes. Then, known amount of potassium nitrate was added, and the solution sample was analyzed in order to obtain a background level of nitrite. The foliage samples were incubated for 2 hours at 20 °C in darkness. Following this procedure, they were given the most optimal conditions for reaction stability.After incubation the amount of synthesized nitrite was determined colorimetrically using sulfanilamide and N-(1-naphthyl)ethylenediamine dihydrochloride, measured at 540 nm. The foliage samples were oven-dried to obtain dry mass. The level of ni-trate reductase activity was calculated as the amount of nitrite produced in nmol per gram of dry mass of foliage tissue per hour. The result obtained during the research demonstrate the changes of nitrate reductase dynamics according to change of incuba-tion parameters. Dynamics of enzyme activity with changes of solution pH and incubation temperature was presented. Installation for conducting infiltrationprocess and construction of in-cubation chamber is also described in this paper

MEASUREMENT OF NITRATE REDUCTASE ACTIVITY IN A FIELD CONDITIONS – METHODOLOGY

During recent three decades interest for biomonitoring and ecological studies was rapidly growing. Therefore was necessary develop of new methods of analysis biochemical parameters whose allow quantify biological response of investigated organisms for environmental factors. The main goal of this paper demonstrates optimal conditions for enzyme kinetics analysis conducted in the field in situ. Nitrate reductase activity is typically assayed in vivo by measuring nitrite production in tissue which has been vacuum infiltrated with buffered nitrate solution. For this study a nitrate reductase assay was adapted from a number of studies with own modifications of authors. Leaves of examined plants were collected on investigated plots and immediately placed into test tubes with buffer solution (potassium phosphate dibasic containing 0.6% propanol-1) and evacuated in 0.33 atm. for 10 minutes. Then known amount of potassium nitrate was added, and the solution sample was analyzed in order to obtain a background level of nitrite. The foliage samples were incubated for 2 hours at 20 °C in darkness. Follow this procedure have given the most optimal conditions for reaction stability. After incubation the amount of synthesized nitrite was determined colorimetrically using sulfanilamide and N-(1-naphthyl)ethylenediamine dihydrochloride, measured at 540 nm. The foliage samples were oven-dried to obtain their dry mass. Level of nitrate reductase activity was calculated as the amount of nitrite produced in nmol per gram of dry mass of foliage tissue per hour. The result obtained during these research demonstrate the changes of nitrate reductase dynamics according to change of incubation parameters. Dynamics of enzyme activity with changes of solution pH and incubation temperature was presented. Installation for conducting infiltration process and construction of incubation chamber is also described in this paper

Lichens and lichenicolous fungi of Magurski National Park (Poland, Western Carpathians)

The paper lists 337 species from Magurski National Park (MNP): 314 lichens, 18 lichenicolous fungi, four saprotrophic fungi and one lichenicolous myxomycete; 112 of them are new for MNP, 75 are reported for the first time for the Beskid Niski Mts, and two are new for Poland. Selected species are accompanied by taxonomic notes and remarks on their distribution in Poland and other Carpathian ranges. First records of Intralichen lichenicola, Burgoa angulosa and Verrucaria policensis and a second record of Epigloea urosperma are given for the whole Carpathian range, and Fuscidea arboricola was recorded for the first time in the Western Carpathians. Halecania viridescens and Mycomicrothelia confusa are new for the Polish Carpathians. The records of Absconditella pauxilla, Collema crispum, Licea parasitica and Rinodina griseosoralifera in MNP are their second known localities for the range. 93 species, mainly rare or threatened in Poland, were reported from MNP in the 20th century but were not refound