Abstract-Lizards (Gallotia galloti) were given either single or consecutive acute oral treatments of the organophosphorus (OP) insecticide parathion in two different experiments. Brain, serum, and liver microsomal esterase activities and liver microsomal monooxygenase activities were measured 6 and 24 h after the single acute treatment at each of four different doses (Experiment 1) or periodically up to 72 d after a number of consecutive acute treatments at two different doses (Experiment 2). Inhibition of serum butyrylcholinesterase (BChE) and carboxylesterase activities was observed in all treatment groups after 24 h and in the groups treated with 2.5, 5, and 7.5 mg/kg of parathion 6 h after treatment. Brain acetylcholinesterase (AChE) was inhibited at all doses after 6 h but only at the highest dose after 24 h. Highly significant nonlinear correlations, based on a piecewise linear regression model, were obtained between brain AChE activity and serum esterase activities at two sampling times after the single acute treatment. Liver microsomal carboxylesterase was found to be induced at the lower doses 6 and 24 h after treatment. Liver microsomal monooxygenase activity was higher 6 h after treatment than at 24 h, but the difference was not statistically significant. In Experiment 2, serum esterase activities recovered exponentially over a period of weeks. An increase in the recovery time to normal esterase activity was observed after each consecutive acute treatment. Brain AChE activity was inhibited at the end of consecutive administrations of parathion at the higher dose, and liver microsomal monooxygenase activity was inhibited at both doses. Symptoms of poisoning were observed in lizards treated with the higher dose of parathion, but no mortality was recorded. Two main conclusions can be drawn: (1) serum esterase activities recovered extremely slowly after acute treatment with parathion and even more slowly after consecutive acute treatments, and (2) there was a nonlinear correlation between the nondestructive biomarkers, serum ''B'' esterases, and the destructive biomarker, brain AChE, 6 and 24 h after exposure. These results suggest that G. galloti should be an ideal bioindicator organism to assess OP exposure in the Canary Islands instead of the birds commonly used for this purpose.
Cryopreservation is currently the only method which allows long-term conservation of living clonal plant material in the vapor or liquid phase of nitrogen (at −140 to −196 °C) allowing tissue to be viable for decades or perhaps centuries. Specifically, for species with recalcitrant seeds or requiring constant vegetative propagation, it is the method of choice for the long-term conservation of its genetic resources. The protocol described here is a modification of a previously developed plant vitrification solution 2 (PVS2)—droplet vitrification method of potato shoot tips, adapted from Musa species. Utilizing this protocol, the International Potato Center (CIP) has successfully stored in the cryobank more than 3000 cultivated potato accessions, belonging to seven species and nine different taxa [16], originating principally from ten countries in South and Central America. As part of CIP’s quality management system, all vegetative material placed in cryo is routinely subsampled, thawed, and assessed to confirm that whole plantlets can be produced after storage in liquid nitrogen. Complete plant recovery rates of thawed shoot tips range from 20% to 100% (average rate: 60%). This chapter describes the complete set of steps from the routine procedure of cryopreserving potato shoot tips for long-term conservation.
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In vitro propagation of Plant Genetic Resources is a basic step for routine genebank and biotechnology research activities. Accelerating growth and rooting of in vitro plants contributes to an improvement in process efficiency and plant quality. In the present study the effect of supplemental thiamine and explant size on biometric variables, ion content in plant sap, chlorophyll content in leaves and moisture content in plants were assessed in a replicated trial on a group of seven in vitro sweetpotato accessions and validated in a set of other 45 accessions. It was shown that adding 0.1 mg L -1 of thiamine to modified Murashige and Skoog culture medium significantly increased plant height, root length, and number of nodes of in vitro sweetpotato shoot culture plants. No significant differences were observed for N0 3 - , K + , Na + and Ca ++ - ion content in plant sap, nor in leaf area, chlorophyll, or moisture content between plants grown with or without thiamine. Uninodal stem segments showed on thiamine-free medium a significantly lower root and plant growth, and reduced number of nodes, than plants grown from uni- and binodal segments on thiamine-supplemented medium. A subsequent experiment tested all the parameters above in a non-replicated screen with a set of 45 diverse sweetpotato accessions. With this diverse set of germplasm, the average plant and root length increased by 41 and 51%, respectively on thiamine-supplemented culture medium compared to the control treatment, confirming that supplemental thiamine is generally beneficial to sweetpotato in vitro shoot culture.
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