Comparative studies of diurnal variations of nitrate reductase activity in wheat, oat and barley

Diurnal variations of in vitro and in vivo (intact tissue assay) nitrate reductase (EC 1.6.6.1) activity and stability were examined in leaves of wheat ( Triticum aestivum L. cv. Runar), oat ( Avcna saliva L. cv. Mustang) and barley ( Hordeum vulgure L. cv. Agneta and cv. Gunillu). Nitrate reductase activity was generally higher for wheat than for oat and barley. However, the diurnal variations of nitrate reductase activity and stability were principally the same for all species, e.g. the high activity during the photoperiod was associated with low stability. All species showed a rapid (30-60 min) increase in the in vitro and in vivo activity when the light was switched on. When light was switched off the in vitro activity decreased rapidly whereas decrease in in vivo activity was slower. These experiments support the hypothesis that an activation/ deactivation mechanism is involved in the regulation of diurnal variations in nitrate reductase activity. Red light enhanced nitrate reductase activity in etiolated wheat and barley leaves. In green leaves, however, the daily increase in nitrate reductase activity was not induced by a brief red light treatment. Indications of different regulation mechanisms for the diurnal variations of nitrate reductase activity among the cereals were not found.     

Light control of extractable nitrate reductase activity in higher plants

Light regulation of extractable nitrate reductase (NR) activity of higher plants is complicated by: 1) involvement of several photoreceptors, 2) differences in the relative importance of the several photoreceptors among species and among developmental stages of the same species, 3) two types of effects – alteration of activity of existing NR and influences on de novo synthesis of NR, and 4) differing forms of NR within the same species. The interrelationships of all of these factors are not clear. It may be that each system will have to be understood separately before a general model can be developed. Immunochemical quantification of NR from systems exposed to varied light regimes may enhance our understanding of this area. Currently few general conclusions can be made; however, we think that the following statements are true or are usually true: (1) Phytochrome influences extractable NR activity by the low irradiance response and high irradiance response in etiolated tissues. (2) In de-etiolated tissues phytochrome can influence NR activity decay at the end of a light period by the low irradiance response. (3) The phytochrome equilibrium or the absolute level of P_fr influences extractable NR activity in green tissues under white light. (4) Blue light influences extractable NR activity through phytochrome and another, unknown, blue light-absorbing pigment. Flavins may be involved in vitro in reactivation of inactivated NR. (5) Photosynthesis does not directly influence the induction of the forms of NR that require substrate and light for induction. (6) In some tissues there appears to be a close link between nitrite-reducing and nitrate-reducing capabilities. (7) Much circumstantial evidence from kinetic and protein-synthesis-inhibitor studies and the only available immunochemical data indicate that light induces de novo synthesis of NR, resulting in increased extractable activity.     

Relationships between macroalgal functional form groups and substrata stability in a subtropical rocky-intertidal system

The general hypothesis that morphological, physiological, and ecological adaptations of macro algal functional-form groups can be related to the level of disturbance encountered in a natural environment was examined. Two articulated calcareous coralline algae (Amphiroa van-bosseae Lemoine, 24% cover and Corallina frondescens Post. & Rupr. 20%) and one non-articulated coralline alga (Lithophyllum sp., 16%), all late-successional predation-tolerant strategists, comprise most of the community cover on stable bedrock substrata at Punta Las Cuevitas, Sonora, Mexico. Conversely, Ulva rigida C. ag. (26% cover) and a ralfsioid crust (23%), shows to be early-successional opportunistic strategists, cover more of the disturbed boulder habitat. Porolithon sonorense Daws., a stress-tolerant strategist, is uniquely abundant on both substratum types (13% cover on boulders, 10% on bedrock). The sheet-like and filamentous algae, prevalent in the temporally unstable habitat, generally show greater productivity (>2×) than the thicker and calcareous forms conspicuous in the more constant environment. It appears that selection for delicate thalli with high productivities, as well as selection for tougher morphologies having lower photosynthetic rates due to greater proportions of structural tissues, are widespread, divergent evolutionary forces among marine algae. Experiments with captive sea urchins (Echinometra vanbrunti Agassiz), in conjunction with fish-preference data published for some of the same algae studied here, offer strong support for the functional-form model. Parrotfishes, rudderfishes, surgeonfishes, damselfishes and E. vanbrunti, in the Gulf of California, preferentially feed on delicate, early-successional, sheet-like, and filamentous algae, while rejecting or ignoring the more structured, late-successional and calcareous algae. There is no significant (P > 0.05) gradation in calorific content between the first four of the six functional groups (i.e., Sheet-, Filamentous-, Coarsely Branched- and Thick Leathery-Groups), but the mean value for these fleshy forms (2.6 kcal · g ash-free dry wt^?1) is significantly greater than that for the last two groups (0.3 kcal, Jointed Calcareous- and Crustose-Groups). The approach used in this study demonstrates a realistic technique for predicting macrophyte community composition from knowledge of the disturbance levels in a given habitat or the reverse. The form group-disturbance relationship has important implications for future biological monitoring of rocky-inter-tidal and subtidal systems.     

An enzyme-probe study of motile domains in the subfragment-2 region of myosin

The temperature-dependence of local melting within the subfragment-2 region of rabbit skeletal muscle myosin has been investigated using an enzyme-probe technique. Rate constants of fragmentation of two long subfragment-2 particles (61,000 Mr and 53,000 Mr per polypeptide chain) and a short subfragment-2 particle (34,000 Mr per polypeptide chain) by three different enzymes (alpha-chymotrypsin, trypsin and papain) have been determined over the temperature range 5 to 40 degrees C. We followed the time-course of digestion at specific sites at high (I = 0.50, pH 7.3) and low (physiological, I = 0.15, pH 7.3) ionic strengths by electrophoresis of the digestion products on sodium dodecyl sulfate-containing gels. All rate constants were corrected for the intrinsic temperature-dependence of the enzymes by comparison with model substrates. Normalized rate constant versus temperature profiles for the three enzyme-probes are similar in showing that local melting in long subfragment-2 (61,000 Mr) occurs in two distinct stages as was observed earlier for the intact myosin rod. Over the temperature range 5 to 25 degrees C a restricted region at Mr = 53,000 to 50,000 from the N terminus of the rod (the light meromyosin/heavy meromyosin junction) shows the highest susceptibility to proteolytic cleavage. At temperatures above 25 degrees C local melting was detected by all three enzymes at several specific sites within the hinge domain (Mr = 53,000 to 34,000). Activation energies for cleavage at the susceptible sites were similar for the three enzyme probes. They suggest that this region of the myosin rod has significantly lower thermal stability than the flanking light meromyosin and short subfragment-2 segments. These results, together with other physico-chemical studies, point to the hinge domain of the myosin cross-bridge as an important functional element in the mechanism of force generation in muscle.     

Structure of tomato bushy stunt virus: V. Coat protein sequence determination and its structural implications

We report the chemically determined sequence of most of the polypeptide chain of the coat protein of tomato bushy stunt virus. Peptide locations have been determined by comparison with the high-resolution electron density map from X-ray crystallographic analysis as well as by conventional chemical overlaps. Three small gaps remain in the 387-residue sequence. Positively charged side-chains are concentrated in the N-terminal part of the polypeptide (the R domain) as well as on inward-facing surfaces of the S domain. There is homology of S-domain sequences with structurally corresponding residues in southern bean mosaic virus.     

Amino acid sequence of rabbit ventricular myosin light chain-2: Identity with the slow skeletal muscle isoform

Many studies have established a correlation of differences in the activities of various muscle types with differences in the expression of myosin isoforms. In this paper we report the sequence determination of myosin light chain-2 from rabbit slow skeletal (LC2s) and ventricular (LC2v) nmscles. We sequenced tryptic peptides from LC2v which account for all except a few terminal amino acid residues. The major part (87 residues) of the rabbit LC2s sequence, obtained from tryptic and cyanogen bromide (CNBr) peptides, was found to be identical to rabbit LC2v. Our results provide the first sequence information on LC2s from any species, and lend strong support to the hypothesis that LC2s and LC2v are identical. Comparisons of rabbit LC2v and LC2s with rabbit LC2f (from fast skeletal muscle), and also with chicken LC2f and LC2v, show clearly that LC2s and LC2v from mammalian and avian species are more closely related to each other than they are to LC2f isoforms from the same species.     

Variation in photosynthetic electron transport capacity in vivo and its effects on the light modulation of ribulose bisphosphate carboxylase

Photosynthetic electron transport capacity was varied in vivo in sugar beets using iron deficiency, and its effects on the light modulation of ribulose bisphosphate carboxylase (RuBPCase) studied. Three treatment groups corresponding to decreasing amounts of thylakoids per leaf area were examined: iron sufficient (control), moderately iron-stressed, and severely iron-stressed. Reduction in electron transport capacity in vivo was correlated with a substantial decrease in the level of RuBPCase activation, even at saturating irradiances. These results indicate a direct relationship between RuBPCase activation and photosynthetic electron transport. In addition, our data suggest that the activation of RuBPCase could not solely account for the increases in the photosynthetic rate at high irradiances; RuBPCase reached maximal activation at irradiances well below light saturation for net photosynthesis.Abbreviations Chl chlorophyll - FeCN ferricyanide - FBPase fructose 1,6-bisphosphatase - RuBP ribulose 1,5-bisphosphate - RuBPCase ribulose 1,5-bisphosphate carboxylase - SBPase sedoheptulose 1,7-bisphosphatase     

Effects of irradiance and temperature on photosynthesis in C3, C4 and C3/C4 Panicum species

Species in the Laxa and Grandia groups of the genus Panicum are adapted to low, wet areas of tropical and subtropical America. Panicum milioides is a species with C₃ photosynthesis and low apparent photorespiration and has been classified as a C₃/C₄ intermediate. Other species in the Laxa group are C₃ with normal photorespiration. Panicum prionitis is a C₄ species in the Grandia group. Since P. milioides has some leaf characteristics intermediate to C₃ and C₄ species, its photosynthetic response to irradiance and temperature was compared to the closely related C₃ species, P. laxum and P. boliviense and to P. prionitis. The response of apparent photosynthesis to irradiance and temperature was similar to that of P. laxum and P. boliviense, with saturation at a photosynthetic photo flux density of about 1 mmol m^-2 s^-1 at 30°C and temperature optimum near 30°C. In contrast, P. prionitis showed no light saturation up to 2 mmol m^-2 s^-1 and an optimum temperature near 40°C. P. milioides exhibited low CO₂ loss into CO₂-free air in the light and this loss was nearly insensitive to temperature. Loss of CO₂ in the light in the C₃ species, P. laxum and P. boliviense, was several-fold higher than in P. milioides and increased 2- to 5-fold with increases in temperature from 10 to 40°C. The level of dark respiration and its response to temperature were similar in all four Panicum species examined. It is concluded that the low apparent photorespiration in P. milioides does not influence its response of apparent photosynthesis to irradiance and temperature in comparison to closely related C₃ Panicum species.Abbreviations AP apparent photosynthesis - I CO₂ compensation point - gl leaf conductance; gm, mesophyll conductance - PPFD photosynthetic photon flux density - PR apparent photorespiration rate - RuBPC sibulose bisphosphate carboxylase     

Redistribution of growth during phototropism and nutation in the pea epicotyl

First positive phototropism of the third internode of intact, 5-d-old pea (Pisum sativum L.) seedlings, grown under continuous, dim red light, showed maximal response following a photon fluence of 3 mol·m^-2 blue light. Greater or lesser fluences (with irradiation time 100 s or less) caused less bending, no response being detectable above 300 or below 0.03 mol·m^-2. Bilateral irradiation with blue light caused no detectable inhibition of growth rate over that range of fluences. The linear nutation of the pea third internode was shown to be driven by a balanced oscillation of growth rate such that the overall growth rate was little changed during the oscillation. Phototropic stimulation changed neither the amplitude nor the period of nutation. Nutation and phototropism probably regulate growth independently. Phototropism in response to the optimal blue light fluence was caused by concomitant depressed growth on the irradiated side and stimulated growth on the shaded side of the bending internode. These results are consistent with the Cholodny-Went hypothesis which states that unilateral blue light induces a lateral redistribution of a growth regulator.Abbreviations R red light - BL blue light Carnegie Institution, Department of Plant Biology paper No. 921     

Blue light induced inhibition of seed germination: The necessity of the fruit coats for the blue light response

The seeds (achenes) of Laportea bulbifera require a chilling to break their dormancy and are negatively photoblastic. Their germination is inhibited by both continuous blue light and continuous or prolonged far-red radiation. The germination of de-coated seeds, prepared by removing the fruit coats, however, was strongly inhibited by continuous far-red, but not by continuous blue light. Photoreversible germination by a brief irradiation with red light occurred when the chilled seeds were exposed to prolonged far-red light. These results suggest that far-red light may regulate the germination of L. bulbifera seeds through the phytochrome system which exists in the regions other than fruit coats and that the blue light reaction may be governed by other photoreceptor system(s).