RESUMO
1. An alkaline pH change occurred when L-rhamnose, L-mannose or L-lyxose was added to L-rhamnose-grown energy-depleted suspensions of strains of Escherichia coli. This is diagnostic of sugar-H+ symport activity. 2. L-Rhamnose, L-mannose and L-lyxose were inducers of the sugar-H+ symport and of L-[14C]rhamnose transport activity. L-Rhamnose also induced the biochemically and genetically distinct L-fucose-H+ symport activity in strains competent for L-rhamnose metabolism. 3. Steady-state kinetic measurements showed that L-mannose and L-lyxose were competitive inhibitors (alternative substrates) for the L-rhamnose transport system, and that L-galactose and D-arabinose were competitive inhibitors (alternative substrates) for the L-fucose transport system. Additional measurements with other sugars of related structure defined the different substrate specificities of the two transport systems. 4. The relative rates of H+ symport and of sugar metabolism, and the relative values of their kinetic parameters, suggested that the physiological role of the transport activity was primarily for utilization of L-rhamnose, not for L-mannose or L-lyxose. 5. L-Rhamnose transport into subcellular vesicles of E. coli was dependent on respiration, was optimal at pH 7, and was inhibited by protonophores and ionophores. It was insensitive to N-ethylmaleimide or cytochalasin B. 6. L-Rhamnose, L-mannose and L-lyxose each elicited an alkaline pH change when added to energy-depleted suspensions of L-rhamnose-grown Salmonella typhimurium LT2, Klebsiella pneumoniae, Klebsiella aerogenes, Erwinia carotovora carotovora and Erwinia carotovora atroseptica. The relative rates of subsequent acidification varied, depending on both the organism and the sugar. L-Fucose promoted an alkaline pH change in all the L-rhamnose-induced organisms except the Erwinia species. No L-rhamnose-H+ symport occurred in any organism grown on L-fucose. 7. All these results showed that L-rhamnose transport into the micro-organisms occurred by a system different from that for L-fucose transport. Both systems are energized by the trans-membrane electrochemical gradient of protons. 8. Neither steady-state kinetic measurements nor binding-protein assays revealed the existence of a second L-rhamnose transport system in E. coli.
Assuntos
Enterobacteriaceae/metabolismo , Fucose/metabolismo , Prótons , Ramnose/metabolismo , Ligação Competitiva , Transporte Biológico Ativo , Metabolismo dos Carboidratos , Carboidratos/farmacologia , Citocalasina B/farmacologia , Eletroquímica , Escherichia coli/metabolismo , Escherichia coli/ultraestrutura , Etilmaleimida/farmacologia , Fucose/farmacologia , Concentração de Íons de Hidrogênio , Cinética , Manose/metabolismo , Manose/farmacologia , Pentoses/metabolismo , Pentoses/farmacologia , Ramnose/farmacologiaRESUMO
A total of 22 grass species were examined from 5 sites spanning the altitudinal range 1550-4350 m.a.s.l. The presence of the C3 or C4 photosynthetic pathway was determined from δ13C values and chilling tolerance was assessed on the basis of electrolyte leakage from leaf slices incubated on melting ice. Most of the grasses studied at the lower altitude sites of 1550 m.a.s.l. (annual mean of daily minimum temperature, 14.6° C) and 2600 m.a.s.l. (9.4° C) possessed C4 photosynthesis and were chill-sensitive. The single except ion was Agrostis avenacea, a montane chill-resistant C3 species which occurred at 2600 m.a.s.l. The three species apparently most sensitive to chilling were Ischaemum polystachyum, Paspalum conjugatum and Saccharum robustum, all occurring at 1550 m.a.s.l. At the higher altitude sites of 3280 (5.6° C), 3580 (4.0° C) and 4350 (-0.7°C) m.a.s.l., most of the grasses exhibited C3 photosynthesis and were chill-resistant. However, an Upland population of the C4 species, Miscanthus floridulus was found at 3280 m.a.s.l. which had acquired chill-resistance as confirmed by additional in vivo variable chlorophyll fluorescence measurements. Cell sap osmotic potential values of the upland grasses at altitudes of 3280-4350 m.a.s.l. were lower (-8.1 to -19.8 bars) than values in grasses from 1550 and 2600 m.a.s.l. (-3.9 to -7.5 bars) due mainly to the presence of non-electrolyte osmoticants, which may be involved in frost avoidance mechanism(s).
RESUMO
The occurrence of Crassulacean acid metabolism (CAM), as judged from δ13C values, was investigated in epiphytes and some related plant species at a series of sites covering the approximate altitudinal range of epiphytes in Papua New Guinea. Comprehensive collections were made at each site and the occurrence of water storage tissue and blade thickness was also determined. Some 26% of epiphytic orchids from a lowland rainforest (2-300 m.a.s.l) showed δ13C values typical of obligate CAM and possessed leaves thicker than 1 mm. A second group of orchids, mostly with succulent leaves, possessed intermediate δ13C values between -23 and -26% and accounted for 25% of the total species number. Some species of this group may exhibit weak CAM or be facultative CAM plants. The remainder of the lowland rainforest species appeared to be C3 plants with δ13C values between -28 and -35%. and generally possessed thin leaves. Obligate CAM species of orchids from a lower montane rainforest (1175 m.a.s.l) comprised 26% of the species total and mostly possessed thick leaves. The remainder of the species were generally thin-leaved with δ13C values between -26 and -35%. largely indicative of C3 photosynthesis. Orchids with intermediate δ13C values were not found in the lower montane rainforest. Obligate CAM appeared to be lacking in highland epiphytes from an upper montane rainforest and subalpine rainforest (2600-3600 m.a.s.l). However the fern, Microsorium cromwellii had a δ13C value of -21.28%. suggesting some measure of CAM activity. Other highland ferns and orchids showed more negative °13C values, up to-33%., typical of C3 photosynthesis. The highland epiphytic orchids possessed a greater mean leaf thickness than their lowland C3 counterparts due to the frequent occurrence of water storage tissue located on the adaxial side of the leaf. It is suggested that low daytime temperatures in the highland microhabitats is a major factor in explaining the absence of CAM. The increased frequency of water storage tissue in highland epiphytes may be an adaptation to periodic water stress events in the dry season and/or an adaptation to increased levels of UV light in the tropicalpine environment.