Residual effects of High Temperature Pre-Treatments on the Germination of Celery Seeds (Apium graveolens)

Celery seeds (Apium graveolens L.) were allowed to imbibe in the dark for different periods at 28. 32. 37 or 41°(the high temperature pre-treatmem or HTP) prior to transfer to 15, 17, 19.5 or 22°C in white light (the low temperature treatment or LTT). The effect of HTP's at 28. 32 and 37°C was to lower the upper temperature limit for germination and this effect increased with increase in the temperature and duration of the HTP. Increasing exposure to an HTP of 41 °C, however, did not appear to lower the upper temperature limit for germination but reduced the viability of the seeds. This trend of increased inhibition of germination caused by increasingly higher temperatures was reversed when the transfer was to a 22°C LTT after a 4 day HTP at 37°C. When the temperature of (he HTP was relatively low and transfer was to a low LTT. or if the HTP was of a brief duration, the rate of germination was increased as compared to seeds not given an HTP. However, when the temperature and duration of the HTP were increased, the rate of germination became slower than that of seeds not given an HTP, i.e. the time taken for the seeds to recover from the HTP-induced dormancy increased with the temperature and duration of the HTP. The decrease in the germination rate was caused by a delay in the start and not a slowing of the rate of embryo elongation within the seed.     

Mutants impaired in vacuolar metal mobilization identify chloroplasts as a target for cadmium hypersensitivity in Arabidopsis thaliana

Cadmium (Cd) is highly toxic to plants causing growth reduction and chlorosis. It binds thiols and competes with essential transition metals. It affects major biochemical processes such as photosynthesis and the redox balance, but the connection between cadmium effects at the biochemical level and its deleterious effect on growth has seldom been established. In this study, two Cd hypersensitive mutants, cad1‐3 impaired in phytochelatin synthase (PCS1), and nramp3nramp4 impaired in release of vacuolar metal stores, have been compared. The analysis combines genetics with measurements of photosynthetic and antioxidant functions. Loss of AtNRAMP3 and AtNRAMP4 function or of PCS1 function leads to comparable Cd sensitivity. Root Cd hypersensitivities conferred by cad1‐3 and nramp3nramp4 are cumulative. The two mutants contrast in their tolerance to oxidative stress. In nramp3nramp4, the photosynthetic apparatus is severely affected by Cd, whereas it is much less affected in cad1‐3. In agreement with chloroplast being a prime target for Cd toxicity in nramp3nramp4, the Cd hypersensitivity of this mutant is alleviated in the dark. The Cd hypersensitivity of nramp3nramp4 mutant highlights the critical role of vacuolar metal stores to supply essential metals to plastids and maintain photosynthetic function under Cd and oxidative stresses.     

To mark the host or the patch: Decisions of a parasitoid searching for concealed host larvae

We found evidence for patch marking in the parasitic wasp Halticoptera laevigata (Hymenoptera: Pteromalidae) foraging for concealed hosts. Wasps attack larvae of the fruit fly Myoleja lucida (Diptera: Tephritidae) in fruits of honeysuckle. A special feature of this host-parasitoid system is the limited food supply of a patch (i.e. a fruit of honeysuckle), which allows the successful development of only a single host fly larva. Females of the parasitoid H. laevigata were found to mark the host patch with a pheromone and to abandon the patch following oviposition into a single host larva. Field data revealed that eggs of the parasitoid were spread out evenly among infested patches, with several larvae of the host fly left unparasitized in those patches that contained more than one host. Since many parasitic insects mark the parasitized host after oviposition, we assumed host marking to be the ancestral character state and studied the patch-marking behaviour of H. laevigata as a derived character state as an alternative foraging strategy. We used stochastic dynamic modelling to investigate under what conditions mutant (patch) markers would be able to invade a population of normal (larval) markers. The models suggested that, under a variety of conditions, wasps marking the patch obtained higher fitness than wasps only marking the larva. Consequently, the results from our model predict the evolution of the patch-marking behaviour found in the empirical investigation. Finally, we discuss alternative pathways to the evolution of patch marking and point out under what circumstances the evolution of a patch-marking behaviour can generally be expected.     

In Vitro Cultivation of the Vascular Phase of Sarcocystis singaporensis

To establish an in vitro culture system for the precystic phase of Sarcocystis singaporensis, we initially tested various excysting fluids for sporocysts. An excysting fluid containing 2.5% bovine taurocholate and 10% bile of the specific intermediate host, Rattus norvegicus, in RPMI medium was the most suitable resulting in excystation of 80% of the sporozoites. Subsequently, we identified brain endothelial cells and pneumonocytes of the rat to promote growth of sporozoites to schizonts. Hepatoma, fibroblastic, or myoblastic cells were not suitable for the parasite's development. First-generation schizonts were seen at days 3-10 postinoculation (PI); a distinct second peak of schizogonic development only occurred in endothelial cells at days 14-18 PI. First-generation schizonts were 26.0 (± 3.8) μm in diameter and contained 32-50 merozoites, second-generation schizonts measured 34.4 (± 10.6) μm and contained 54-72 merozoites. Merozoite yield at large-scale culture conditions (75 cm² flasks) using pneumonocytes as host cells was relatively low. Ultrastructurally, sporozoites and merozoites were quite similar to corresponding stages of other Sarcocystis species. With regard to host cell specificity and developmental kinetics, in vitro cultivation showed close similarities to the situation in vivo.     

An Electron Microscope-Cytochemical Method for Differentiating Membranes of Host Red Cells and Malaria Parasites*

Limiting membranes of malaria parasites and host red cells stain differently when exposed to positively charged iron colloid. Negatively charged red cell membranes avidly bind colloid, whereas parasite membranes do not. This selectivity in colloidal iron uptake by the 2 types of membranes can be utilized as an aid in discerning the amounts of contaminating host cell membranes in “free” malaria parasite preparations and in related cell-free membrane extracts.