Protein Uptake and Digestion in Bloodstream and Culture Forms of Trypanosoma brucei*

SYNOPSIS. The mechanisms of ferritin uptake and digestion differ in bloodstream and culture forms of Trypanosoma brucei. Ferritin enters bloodstream forms from the flagellar pocket by pinocytosis in large spiny-coated vesicles. These vesicles become continuous with straight tubular extensions of a complex, mostly tubular, collecting membrane system where ferritin is concentrated. From the collecting membrane system the tracer enters large digestive vacuoles. Small spiny-coated vesicles, which never contain ferritin, are found in the Golgi region, fusing with the collecting membrane system, and around the flagellar pocket. Acid phosphatase activity is present in some small spiny-coated vesicles which may represent primary lysosomes. This enzymic activity is also found in the flagellar pocket, pinocytotic vesicles, the collecting membrane system, the Golgi (mature face), and digestive vacuoles of bloodstream forms. About 50% of the acid phosphatase activity of blood forms is latent. The remaining nonlatent activity is firmly cell-associated and probably represents activity in the flagellar pocket. The structures involved in ferritin uptake and digestion are larger and more active in the short stumpy than in the long slender bloodstream forms. The short stumpy forms also have more autophagic vacuoles. No pinocytotic large, spiny-coated vesicles or Golgi-derived, small spiny-coated vesicles are seen in culture forms. Ferritin leaves the flagellar pocket of these forms and enters small smooth cisternae located just beneath bulges in the pocket membrane. The tracer then passes through a cisternal collecting membrane network, where it is concentrated, and then into multivesicular bodies. In the culture forms, acid phosphatase activity is localized in the cisternal system, multivesicular bodies, the Golgi (mature face), and small vesicles in the Golgi and cisternal regions. The flagellar pocket has no acid phosphatase activity, and almost all the activity is latent in these forms. The culture forms do not release acid phosphatase into culture medium during 4 days growth. Uptake of ferritin by all forms is almost completely inhibited by low temperature. These differences among the long slender and short stumpy bloodstream forms and culture forms are undoubtedly adaptive and reflect different needs of the parasite in different life cycle stages.     

Thick Filaments in Unstretched Mammalian Smooth Muscle

THE controversy concerning the organization of myosin in mammalian smooth muscle was reviewed (Nature New Biology, 231, 225; 1971) at a time when the studies of Rice's laboratory and our own demonstrated a regular, quasi-rectangular array of thick filaments in guinea-pig taenia coli (TC) and rabbit portal-anterior mesenteric vein (MV), and, further, that, by excessive stretch and by the use of hypertonic incubation solutions, the thick filaments in this lattice could be aggregated into ribbon-like structures^1,2. These observations were made on muscles stretched to approximately 1.5 times their excised length. Both the TC³ and the rabbit MV^2,4 are spontaneously active smooth muscles, which shorten to less than their in vivo length when excised from the body: stretching by approximately 1.5 times brings these muscles close to their in vivo length. Nevertheless, recent reports^5,6, indicating that thick filaments were more readily visualized (but see Figs. 2 and 3 in ref. 5) in stretched smooth muscles, prompted the editorial writer of Nature (231, 423; 1971) to consider it a debatable question whether thick filaments are present in unstretched muscle. Thick filaments have been observed in relaxed muscles^1,5,6 and we now show that an array of thick filaments can also be observed in completely unstretched guinea-pig and rabbit MV smooth muscle (excised and dropped into the fixative) and that such arrays are present after two different modes of fixation.     

The reproductive biology of a distylous tree, Sarcotheca celebica (Oxalidaceae) in Sulawesi, Indonesia

Sarcotheca celebica is a tree endemic lo Sulawesi (and Kabaena Island). Kike many species in the Oxalidaceae it is heterostylous (here distylous). The small red flowers, which last less than one day, are produced in loose inflorescences which bloom over a long flowering season (several months). There are two whorls of stamens in each plant, which overlap in length, perhaps indicating a tristylous origin. Long–styled and short–styled plants differed in the measurements of stamens, styles and pollen grains. Only a small amount of pollen was produced by long–styled plants. Effective insect pollinators were mainly large Hymenoptcra, especially Xylocopa species, which visited many flowers and different trees in rapid succession. A wide diversity of insects visited flowers of both morphs, and numbers visiting each morph were approximately equal. The only successful experimental pollinations were from pollen of short–styled plants onto stigmas of long–styled plants. Automatic self–pollination did not occur. In nature not all flowers produced fruit, some fruits were set with no seeds and a low number of seeds was set in all fruits. The differences in seed set per fruit between short–styled, long–styled and experimental crosses indicate that pollinator visits were insufficient for maximum seed set in some fruits, but selective abortion of seeds must also be occurring. We suggest that S. celebica may represent an intermediate stage in the evolution of dioecy from distyly, with the short–styled flowers making their major genetic contribution through pollen and long–styled flowers through ovules.     

Flow Cytometric Analysis of Immunoglobulin and Complement Component C3 on the Surface of Trypanosoma lewisi1

ABSTRACT. We have reinvestigated whether surface immunoglobulin (sIg) on Trypanosoma lewisi is antibody directed toward parasite antigen by using flow cytometry to analyze parasites stained with fluoresceinated F(ab′)₂ fragments of antibodies to rat IgG and IgM. We have confirmed that IgG antibody to the parasites is present both in the serum of rats and on the surface of parasites between the fourth and twentieth days of infection, that the amount of sIg per cell increases as the infection progresses, and that considerably more IgG is present on parasites harvested from intact rats than on those from rats that had been immunosuppressed by whole body γ-irradiation. In addition sIgM was detected on trypanosomes from intact, but not on parasites from irradiated rats. We have also made two observations suggesting that not all sIg is specific antibody made in response to T. lewisi. First, a low but significant amount of sIgG was detected on parasites throughout infection in irradiated rats; no sIgM was detected on these parasites. Second, when parasites harvested from immunosuppressed rats were incubated in normal rat serum, the amount of both sIgG and sIgM detected by flow immunofluorescence increased. Parasites harvested from intact animals bound IgM but not IgG from normal rat serum. These results suggest either that natural antibody to the trypanosomes is present in the serum of uninfected rats or that some rat immunoglobulins bind to structures on the trypanosome surface in ways that do not depend on usual antigen-antibody interactions. Finally, flow immunofluorescence was also used to detect complement component C3 on the surface of both intact and trypsinized bloodstream forms harvested from intact or immunosuppressed rats. The amount of sC3 per cell did not increase until late in the infection and consequently did not correlate with the increase of sIgG. Therefore, T. lewisi avoids destruction by the immune system although immune effector molecules, IgG, IgM, and C3, are on its surface.     

Gregarious flowering in a lowland tropical rainforest: A possible response to disturbance by Cyclone Winifred

Observations in tropical lowland rainforests damaged by Cyclone Winifred on 1 February 1986 suggest that the disturbance has been followed by an unusual burst of flowering. Fifty-four species of concurrently flowering shrubs and trees were recorded in early March along a 4 km road transect in a cyclone-damaged forest. Approximately 80% of the smaller tree and shrub species observed were flowering. Twelve species which had flowered and fruited prior to the cyclone flowered again. Some species which normally flower following periods of partial or complete deciduousness in the drier months of the year flowered after being defoliated by the cyclone. Widespread defoliation of the rainforests, together with the hot, dry weather which followed the cyclone, exposed the understorey layers to abrupt increases in temperature, insolation, and water stress. It is postulated that these environmental changes may have effected the synchronization of flowering of a larger number of species than would normally occur. It is suggested that longterm changes in the species composition of the damaged forests as a result of coincidences of seed availability and regenerative space may occur.     

Effects of Temperature and dl-Strigol on Seed Conditioning and Germination of Witchweed (Striga asiatica)

Seed conditioning and germination in witchweed (Striga asiatica(L.) Kuntze) were temperature-dependent. With higher conditioningtemperatures, shorter conditioning time was required for germinationwith terminal dl-strigol (strigol) treatment at 30 °C. Maximumgermination (80–100%) was obtained by conditioning inwater at 20, 25, 30 and 35 °C for 14, 7, 5 and 3 d, respectively,and terminally treating with 10^–6 M strigol at 30 °C.Seeds conditioned in 10^–8 M strigol instead of water germinatedmuch less with the same terminal strigol treatment. Generally,conditioning was slower when seeds were conditioned in strigolrather than water. The reduction in germination rate by pretreatmentin strigol or pretreatment at low temperatures could be overcomeby increasing the terminal strigol concentration in the germinationtest. Conditioned seeds did not germinate at 10 and 15 °Cwith a terminal 10^–6 M strigol treatment but yielded closeto maximum germination at 25, 30 and 35 °C with the sameterminal strigol treatment. To obtain maximum germination, boththe minimum conditioning temperature and the minimum germinationtemperature for conditioned seeds were 20 °C. Factors suchas conditioning time, and strigol concentration and temperatureduring conditioning and/or germination determine whether seedsremain in the conditioning phase or shift to a germination phase. dl-Strigol, germination stimulation, parasitic plants, seed conditioning, seed germination, Striga asiatica, temperature, weed control     

Dispersal ability and genetic structure in aquatic invertebrates: a comparative study in southern California streams and reservoirs

1. The natural seasonal drying and flooding of southern California streams have been altered over the past century by activities related to agriculture, flood control, and reservoir construction. The genetic structure and diversity of aquatic invertebrates inhabiting these environments is largely unexplored, and may be important for conservation. 2. We sampled two species of aquatic invertebrates with different dispersal abilities to assess genetic structure and diversity, and make inferences about the evolutionary processes that underlie these genetic patterns. The mayfly Fallceon quilleri, which has a winged terrestrial stage, was sampled from perennial and intermittent streams from three catchments across San Diego County. The amphipod Hyalella azteca was sampled from streams (perennial and intermittent) and reservoirs in a single catchment (San Dieguito). Because it is obligately aquatic throughout its life‐cycle, H. azteca was assumed to disperse less than F. quilleri. 3. Intrapopulation and overall genetic diversity was higher in F. quilleri than in H. azteca. In F. quilleri there was very little genetic divergence among populations, and most of the genetic differentiation that was observed could be attributed to a single population. In H. azteca, populations were markedly differentiated between the upper and lower segments of the San Dieguito basin, which are separated by a c. 10 km section of stream that rarely has surface flow. Within both segments, genetic divergence between sites connected by reservoirs and perennial streams was not significantly different. 4. Our results suggest that F. quilleri disperses widely and thus avoids genetic bottlenecks and marked levels of population differentiation that may be expected from frequent extinctions and recolonizations. In contrast, restricted dispersal in H. azteca is associated with relatively low genetic diversity and high genetic divergence across a portion of the catchment in which surface flow is rare.     

Genome evolution in allotetraploid Nicotiana

The nuclear cytoplasmic interaction (NCI) hypothesis of genome evolution and speciation in plants states that newly formed allopolyploids pass through a bottleneck of sterility and the fertile plants that emerge are fixed for species‐specific chromosome translocations. These translocations restore fertility and reduce negative effects of the maternal cytoplasm on an alien paternal genome. Using fluorescent in situ hybridization and genomic in situ hybridization and by reviewing published data, we test the NCI hypothesis using three natural Nicotiana allotetraploids (all 2n = 4x = 48, N. arentsii, N. rustica and several genotypes, including a feral plant and cultivars, of N. tabacum (tobacco)). We compare these data with three synthetic tobacco plants (Th37) that are F3 descendent progeny of an allotetraploid formed from ♀N. sylvestris (2n = 24) ×♂N. tomentosiformis (2n = 24). No intergenomic translocations were observed in N. arentsii and N. rustica. An analysis of subtelomeric tandem repeats in these allotetraploids and their putative parents shows minimal genetic changes; those that do occur may reflect evolution in the diploids or the polyploids subsequent to allopolyploidy. All natural N. tabacum genotypes have intergenomic translocations. This may reflect a large ‘genomic‐shock’ generated by allopolyploidy involving widely diverged parental species. Two of three synthetic tobacco plants had a translocation similar to that found in all cultivars of tobacco. This translocation may be significant in tobacco fertility and may have been fixed early in tobacco's evolution. But it is lacking in the feral tobacco, which might indicate a polyphyletic origin or early divergence from all cultivars examined. Overall, only in tobacco is there any evidence that NCI may have influenced genome evolution, and here further data are required to verify chromosome identity. © 2004 The Linnean Society of London, Biological Journal of the Linnean Society, 2004, 82 , 599–606.