In Vitro Propagation of the Protozoan Perkinsus Marinus, A Pathogen of the Eastern Oyster, Crassostrea Virginica

ABSTRACT. Perkinsus marinus , a pathogen of eastern oysters ( Crassostrea virginica ), has been successfully propagated in vitro. Cultures of the parasite were initiated from heart fragments of an infected oyster. the cultured protozoan (designated Parkinsus -1) was similar in morphology at both the light and transmission electron microscopy levels to histozoic stages of P. marinus in naturally infected oysters. In addition, cultured cells incubated in fluid thioglycollate medium produced enlarged cells (prezoosporangia) that stained blue-black in Lugol's solution, a response characteristic to Perkinsus spp. and used in routine diagnosis. Polyclonal antibodies raised against P. marinus prezoosporangia reacted positively to Perkinsus -1. Finally, the cultured cells infected susceptible oysters and reisolation of Perkinsus -1 cells was possible from the hearts of experimentally infected oysters. the culture medium contained most of the known constituents of cell-free hemolymph of oysters. the success achieved in culturing P. marinus will allow further investigations aimed at reducing mortalities caused by this important oyster pathogen and at addressing many unanswered questions about its biology and pathobiology.     

Plant Chromatin Replicated in the Absence of Protein Synthesis

The aim of the present work is to detect possible differencesin the chromatin of plants replicated in the absence of proteinsynthesis. The kinetics of nuclease digestion in Allium cepa L., evaluatedafter making the cells permeable, was faster for the chromatinof meristem cells replicated in the presence of 1.0 µgml^–1 cycloheximide than in control cells. In order to have a synchronous population in the meristems,cells were labelled as binucleate by a short treatment with5.0 mM caffeine. Treated cells failed to increase both theircontent in dense chromatin and intranuclear histones. Thesefacts suggest that chromatin replicated in the presence of cycloheximidedid not incorporate histones and was unable to be integratedinto dense chromatin patches. Key words: Allium cepa L., Chromatin replication, Protein synthesis     

Radiation-induced improvement of the freeze-resistance of a cold-sensitive wheat variety: Freeze-tolerance and fatty acid patterns

Effects of 3–15 krad ⁶⁰Co gamma radiation on cereal seeds were investigated with regard to the frost hardiness of leaves of 5–7-day-old seedlings. Comparative studies were carried out on the gas-chromatographically determined distribution patterns of fatty acids in different pools (total fatty acid, total lipid, polar lipids) of a cold-resistant (cv. Mironowskaya 808) and a cold-sensitive (cv. Penjamo 62) cultivar of wheat (Triticum aestivum L.). Parallelism between fatty acid distribution pattern, empirical parameters suggested for quantitative measures of cold tolerance (the ratio unsat/sat and double-bond index), and the low-temperature behaviour of shoots grown from radiation treated seeds of ‘Penjamo 62’, was also examined. To monitor differences in the fatty acid syntheses of ‘Mironowskaya 808’and ‘Penjamo 62’, and to demonstrate radiation-induced changes in fatty acid turnover a [1–¹⁴C]-acetate incorporation technique was employed. The results of practical importance are: 1. A significant improvement in the frost-resistance of the cold-sensitive ‘Penjamo 62’variety could be achieved with 6–9 krad irradiation, the half-freeze-killing temperature dropping from ?6 to about ?18°C. 2. Freeze-hardiness, no matter whether inherited or gained, could be abolished by gamma irradiation with higher dose. The following conclusions of theoretical interest can be drawn: 1. Low-temperature behaviour of plant tissues is a dynamic property rather than a static one, not only the formation but also the breakdown of certain hydrocarbon chains may be of paramount relevance in cold-tolerance. 2. Not the linolenic (18:3) component, but rather the overall distribution pattern of the C₁₈ family seems to correlate with low-temperature-responses of shoots. 3. Empirical parameters investigated do not provide reliable quantitative measures of the susceptibility to freeze (and cold-) injuries. 4. Changes in the syntheses of some key proteins (e.g. peroxidase, water-soluble proteins, etc.) may also be important in adaptation to low-temperature conditions.     

A new Spanish species of Tragopogon (Asteraceae: Lactuceae)

Tragopogon cazorlanum (Asteraceae: Lactuceae) is proposed as a new Spanish species with distribution restricted to the Baetic mountains (south-eastern Spain). Its morphological, palynological, chromosomal, biogeographical and ecological features are discussed, as well as its main relationships and differences with reference to other Spanish species of the genus.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 146 , 505–511.