Far-red light (FR) inhibition of seed germination of tomato (Solanum lycopersicum L.) was studied with the phytochrome (phy)-hypersensitive mutants, hp-1w, hp-1w,fri1, a phyA-deficient double mutant, and hp-1w,tri1, a phyB1-deficient double mutant. Seeds of all mutants germinated readily in the dark at 25 degrees C, and the germination was retarded by a single 100-s FR pulse given 1-3 h after sowing. The effect of an FR pulse was red-light reversible in all mutants used. After 24 h where a single FR pulse was no longer effective, prolonged FR exposure or hourly FR pulses suppressed germination in hp-1w and hp-1w,tri1, whereas in hp-1w,fri1 the suppressive effect of FR was almost absent. The effect of the prolonged FR was greater than that of the hourly 3-min FR pulses having equal photon fluence, and was fluencerate dependent. Thus we conclude that the germination inhibition by FR in tomato seed consists of a low-fluence response and a high irradiance response (HIR); the latter is controlled by phyA, but not phyB1. This is the first indication of phyA being involved in the HIR of seed germination inhibition.
Abstract. Anthocyanin synthesis in the broom sorghum,Sorghum bicolor Moench cvs. Acme Broomcorn and Sekishokuzairai-Fukuyama, is mediated separately or synergistically by an ultraviolet light-B (UV-B) photoreceptor and phytochrome. When seedlings were exposed to moderate low temperatures ranging from 12 to 20 ~ C before irradiation, only the phytochrome-mediated anthocyanin synthesis was markedly enhanced compared with the control, which was kept throughout at 24 ~ C; synthesis mediated by the UV-B photoreceptor was unaffected. The effectiveness of an exposure to 20 ~ C increased as the duration of exposure increased up to 24 h and as the time of exposure became closer to the time of irradiation. However, when seedlings were exposed to 20~ from after irradiation until harvest, anthocyanin syntheses induced by both UV-B and red light were equally suppressed, probably due to the general reduction of metabolism involved in anthocyanin synthesis that is a consequence of lower temperature. The results support the view that the signal transduction of the pyhtochrome system is different from that of the UV-B photoreceptor, and indicate that the phytochrome system may involve a step or steps which are amplified by a previous exposure to the moderate low temperature.
The supply of phosphorus, the essential element for plant growth and development, is often limited in natural environments. Plants employ multiple physiological strategies to minimize the impact of phosphate deficiency. In deciduous trees, phosphorus is remobilized from senescing leaves in autumn and stored in other tissues for reuse in the following spring. We previously monitored the annual changes in leaf phosphate content of white poplar (Populus alba) growing under natural conditions and found that about 75 % of inorganic and 60 % of organic leaf phosphates observed in May were remobilized by November. In order to analyze this process (such annual events), we have established a model system, in which an annual cycle of phosphate re-translocation in trees can be simulated under laboratory conditions by controlling temperature and photoperiod (='shortened annual cycle'). This system evidently allowed us to monitor the annual changes in leaf color, phosphate remobilization from senescent leaves, and bud break in the next spring within five months. This will greatly facilitate the analysis of cellular and molecular mechanisms of annual phosphate re-translocation in deciduous trees.
Seasonal recycling of nutrients is an important strategy for deciduous perennials. Deciduous perennials maintain and expand their nutrient pools by the autumn nutrient remobilization and the subsequent winter storage throughout their long life. Phosphorus (P), one of the most important elements in living organisms, is remobilized from senescing leaves during autumn in deciduous trees. However, it remains unknown how phosphate is stored over winter. Here we show that in poplar trees (Populus alba L.), organic phosphates are accumulated in twigs from late summer to winter, and that IP6 (myo-inositol-1,2,3,4,5,6-hexakis phosphate: phytic acid) is the primary storage form. IP6 was found in high concentrations in twigs during winter and quickly decreased in early spring. In parenchyma cells of winter twigs, P was associated with electron-dense structures, similar to globoids found in seeds of higher plants. Various other deciduous trees were also found to accumulate IP6 in twigs during winter. We conclude that IP6 is the primary storage form of P in poplar trees during winter, and that it may be a common strategy for seasonal P storage in deciduous woody plants.
Contrary to the established notion that the apical hook of dark-grown dicotyledonous seedlings opens in response to light, we found in tomato (Solanum lycopersicum L.) that the apical hook curvature is exaggerated by light. Experiments with several tomato cultivars and phytochrome mutants, irradiated with red and far-red light either as a brief pulse (Rp, FRp) or continuously (Rc, FRc), revealed: the hook-exaggeration response is maximal at the emergence of the hypocotyl from the seed; the effect of Rp is FRp-reversible; fluence-response curves to a single Rp or FRp show an involvement of low and very low fluence responses (LFR, VLFR); the effect of Rc is fluence-rate dependent, but that of FRc is not; the phyA mutant (phyA hp-1) failed to respond to an Rp of less than 10(-2) micromol m(-2) and to an FRp of all fluences tested as well as to FRc, thus indicating that the hook-exaggeration response involves phyA-mediated VLFR. The Rp fluence-response curve with the same mutant also confirmed the presence of an LFR mediated by phytochrome(s) other than phyA, although the phyB1 mutant (phyB1 hp-1) still showed full response probably due to other redundant phytochrome species (e.g., phyB2). Simulation experiments led to the possible significance of hook exaggeration in the field that the photoresponse may facilitate the release of seed coat when seeds germinate at some range of depth in soil. It was also observed that seed coat and/or endosperm are essential to the hook exaggeration.
A comprehensive analysis of the levels of primary metabolites in wild type (WT) and several auxin-signaling mutants namely, tir1, slr and arf7 arf19 of Arabidopsis thaliana has been performed using CE-MS, a technique particularly sensitive for the measurement of polar compounds. We first measured the levels of primary metabolites in shoots and roots, most of the analyzed metabolites were found to be quantitatively and qualitatively comparable in WT and three kinds of mutants (tir1, slr and arf7 arf19). Some amino acids such as GABA, Arg, Orn, Val, Thr, Leu and Ile exhibited a unique pattern of distribution between shoots and roots in both WT and the mutants. On the other hand, the mutant slr showed a quite different pattern of metabolites measured in the present study.Subsequently, the responses of primary metabolites to a short-term (60 min) application of exogenous IAA (10 −7, 10 −8 M) in WT and the mutants were characterized. Due to IAA treatments, some amino acids such as GABA in WT roots and Gly and Ala in WT shoots were altered, but not in the mutants. Gln was altered in slr shoots by 10 −7 M IAA treatment. Levels of G6P from the glycolic pathway were altered in WT roots and those of 2PG, 3PG were altered in tir1 shoots in response to IAA treatments. The levels of succinate in TCA cycle were altered by IAA treatments in WT shoots but not in the mutants. IAA treatment inhibited the respiration in WT roots. The suppression of respiration might account for the IAA-dependent alteration of some metabolites. Difference of auxin responses between WT and auxin-signaling mutants suggests that some metabolic processes are under IAA control.
We analyzed the metabolites and proteins contained in pure intact vacuoles isolated from Arabidopsis suspension-cultured cells using capillary electrophoresis-mass spectrometry (CE-MS), Fourier transform-ion cyclotron resonance (FT-ICR)-MS and liquid chromatography (LC)-MS. We identified 21 amino acids and five organic acids as major primary metabolites in the vacuoles with CE-MS. Further, we identified small amounts of 27 substances including well-known vacuolar molecules, but also some unexpected substances (e.g. organic phosphate compounds). Non-target analysis of the vacuolar sample with FT-ICR-MS suggested that there are 1,106 m/z peaks that could predict the 5,090 molecular formulae, and we have annotated 34 compounds in these peaks using the KNapSAck database. By conducting proteomic analysis of vacuolar sap, we found 186 proteins in the same vacuole samples. Since the vacuole is known as a major degradative compartment, many of these were hydrolases, but we also found various oxidoreductases and transferases. The relationships between the proteins and metabolites in the vacuole are discussed.
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