Low Fluence Response Research Articles

Changes in the light sensitivity of buried Polygonum aviculare seeds in relation to cold‐induced dormancy loss: development of a predictive model

The effect of cold (stratification) temperature on changes in the sensitivity of Polygonum aviculare seeds to light was investigated. Seeds buried in pots were stored under stratification temperatures (1.6, 7 and 12 degrees C) for 137 d. Seeds exhumed at regular intervals during storage were exposed to different light treatments. Germination responses obtained for seeds exposed to different light treatments and stratification temperatures were used to develop a model to predict the sensitivity of buried seeds to light. Seed sensitivity to light increased as dormancy loss progressed, showing the successive acquisition of low-fluence responses (LFR), very low-fluence responses (VLFR), and the loss of the light requirement for germination for a fraction of the seed population. These changes were inversely correlated to stratification temperature, allowing the use of a thermal time index to relate observed changes in seed light sensitivity to stratification temperature. The rate of increase in sensitivity of P. aviculare seeds to light during stratification is inversely correlated to soil temperature, and these changes in light sensitivity could be predicted in relation to temperature using thermal-time models.

Circumnutation of rice coleoptiles: its occurrence, regulation by phytochrome, and relationship with gravitropism

It has been found that coleoptiles of dark-grown rice (Oryza sativa L.) seedlings undergo regular circumnutation in circular orbits with periods of about 180 min. Both clockwise and counter-clockwise movements were observed, but individual coleoptiles continued to rotate only in one direction. Light-grown seedlings did not show circumnutation. In fact, dark-grown seedlings were found to cease circumnutating in response to a pulse of red light (R). This light-induced inhibition of circumnutation was demonstrated to involve both a FR-inducible very-low-fluence response, solely mediated by phytochrome A, and a FR-reversible low-fluence response, mediated by phytochrome B and/or C. The R-induced inhibition of circumnutation showed temporal agreement with the R-induced inhibition of coleoptile growth, suggesting that the former results from the latter. However, about 25% of growth activity remained after R treatment, indicating that circumnutation is more specifically regulated by phytochrome. The R-treated coleoptile showed gravitropism. Investigation of the growth differential for gravitropic curvature revealed that gravitropic responsiveness was rather enhanced by R. The results suggested that gravitropism is not a cause of circumnutation. It remained probable, however, that gravity perception is a part of the mechanism of circumnutation. It is speculated that the circumnutation investigated aids the seedling shoot in growing through the soil.

Characterization of sunlight-grown transgenic rice plants expressing Arabidopsis phytochrome A

Transgenic rice (Oryza sativa) overexpressing Arabidopsis phytochrome A (phyA) was cultivated up to the T3 generation in paddy to elucidate the role of phyA in determining the plant architecture and the productivity of sunlight-grown rice plants. PhyA is light-labile and controls plant growth in response to the far-red light-dependent high-irradiance response as well as the very low fluence response. The Arabidopsis phyA gene linked to the rice rbcS promoter was transformed into embryogenic rice calli, and the calli were regenerated to whole plants. Compared to wild-type seedlings, the rbcS::PHYA transgenic seedlings contained more phyA when grown in the dark, and at least 10-fold more phyA when exposed to white light. When grown in paddy, the phyA transgenic plants in general exhibited reduced plant height (dwarfing), larger grain size, higher chlorophyll content, smaller tiller number, and low grain fertility compared to wild-type plants. The heading stage was not significantly changed. However, it is likely that a certain level of phyA is a prerequisite for induction of such changes. It is suggested that phyA overproduction in rice could be a useful tool to improve rice grain productivity by the larger grain size that increases grain yield and the dwarfing that tolerates lodging-associated damage.

Role of marble microstructure in near-infrared laser-induced damage during laser cleaning

When marble is cleaned by nanosecond neodymium yttrium–aluminum–garnet lasers (1064 nm), strongly absorbing surface contaminants are removed at fluences substantially below the damage threshold for the much less absorptive marble substrate. Recent studies have shown, however, that unacceptable roughening of the marble surface also may occur at low fluences due to removal of individual grains. In order to elucidate this effect, we have compared the low-fluence response of marbles with two different grain sizes and single-crystal calcite, in the fluence range 0.12–1.25 J cm−2. Damage was greater in fine-grained than coarse-grained marble, and did not occur in the single-crystal calcite at these fluences. The temperature rise following defect-mediated absorption triggers thermal plasma emission and generates shock waves; the concomitant surface damage depends on the size and crystallographic orientation of the crystals. Laser irradiation anneals the defects and increases “crystallite size.” The implications for the laser-assisted cleaning of marble artworks are outlined.

Expansin gene expression in Datura ferox L. seeds is regulated by the low-fluence response, but not by the high-irradiance response, of phytochromes

Expansins are a multi-gene family of proteins involved in changes in cell wall properties. In seeds where the embryo is completely surrounded by the endosperm, dormancy breakage requires weakening of the micropylar endosperm, increased embryo growth potential or both. Cell wall alterations are fundamental components of these processes, and expansins are thought to participate in them. Here, we explore the possible involvement of expansins in the control of germination by phytochrome in Datura ferox L. seeds. Based on the conserved sequences of known expansin genes, corresponding primers were designed to investigate the expression of expansin mRNAs by reverse transcriptase polymerase chain reaction (RT-PCR). One expansin mRNA was detected in micropylar endosperm of D. ferox, while two mRNAs were present in the embryo prior to radicle emergence. Expansin transcript content is promoted by red (R) light, both in the micropylar endosperm and the embryo; these effects of R are far-red light (FR) reversible, displaying a typical low-fluence response (LFR) in a way fully consistent with the photocontrol of germination. On the other hand, when the promotion of germination by the LFR is antagonized by exposing the seeds to continuous FR through a high-irradiance response (HIR) or by ABA, the inhibitory treatments do not affect expansin mRNA abundance. The results support the participation of expansins in the promotion of germination by LFR of phytochromes, and suggest that inhibition of germination by HIR or ABA does not include a reduction in the transcription of expansin genes.

Blue light-induced chloroplast reorientations in Lemna trisulca L. (duckweed) are controlled by two separable cellular mechanisms as suggested by different sensitivity to wortmannin.

Chloroplast reorientations within mesophyll cells are among the most rapid physiological responses of higher plants to blue light. At light intensities below the saturation point of photosynthesis, chloroplasts move to the cell walls perpendicular to the direction of light and maximize light absorption (low-fluence rate response [LFR]). At light intensities above the saturation point of photosynthesis, chloroplasts redistribute to cell walls parallel to the direction of light (high-fluence rate response [HFR]). The actin-based mechanism is responsible for the light-induced chloroplast movements. We have found that an inhibitor of phosphoinositide-3-kinases, wortmannin, potently and irreversibly inhibited LFR and HFR chloroplast responses to blue light in Lemna trisulca L. mesophyll cells. Microscopic observations and photometric measurement indicated that 100 nM wortmannin specifically inhibited LFR in Lemna, whereas HFR displayed no sensitivity to the inhibitor at this concentration. A complete inhibition of the HFR could be obtained by 1 microM wortmannin. These data indicate that LFR is more sensitive to wortmannin than HFR and suggest that these two responses may be under the control of different cellular mechanisms. Our results suggest that phosphoinositide kinases and other phosphoinositide cycle enzymes may play a role in the transduction of the light signal to the actin cytoskeleton in Lemna as factors specifying the direction of chloroplast movements. A hypothetical model assuming three signaling pathways regulating light-induced chloroplast reorientations in mesophyll cells is proposed.

Are NADP-dependent isocitrate dehydrogenases and ferredoxin-dependent glutamate synthase co-regulated by the same photoreceptors?

Activities of NADP-dependent isocitrate dehydrogenases (cytosolic and plastidic isoforms, ICDH1 and ICDH2; EC 1.1.1.42) and ferredoxin-dependent glutamate synthase (Fd-GOGAT; EC 1.4.7.1) in turions of Spirodela polyrhiza were all stimulated by light. Single or repeated red light (R) pulses induced the activity of the enzymes and this effect was reverted by subsequent far-red light (FR) pulses. The enzymes are, therefore, co-regulated by the low-fluence response of phytochrome. For ICDH, this is reported here for the first time. Neither an effect of the very low-fluence response nor of the FR-mediated high-irradiance response was detectable. Irradiance with continuous R resulted in enhanced enzyme activities and protein levels (Western analysis using polyclonal antibodies against ICDH1 and Fd-GOGAT). These additional effects of continuous R (called a "non-induction effect") could be inhibited for ICDH1 and ICDH2 by the inhibitor of photosynthetic electron transport, 3-(3',4'-dichlorophenyl)-1,1-dimethylurea, and are therefore related to the effect of photosynthesis. In contrast, the non-induction effect of Fd-GOGAT was resistant against this inhibitor. Moreover, hourly R pulses did not replace the effect of continuous R. The non-induction effect of light on the activity and protein level of Fd-GOGAT was therefore tentatively classified as an R-mediated high-irradiance response. The activity of Fd-GOGAT but not that of ICDHs was additionally regulated by a specific blue-light receptor. It can be concluded that the levels of ICDHs and Fd-GOGAT were coordinated by light but were not co-regulated by the same photoreceptors. Nitrate is necessary for the light regulation of both enzymes, contributing to the coordinated expression of the relevant genes.

A growth regulatory loop that provides homeostasis to phytochrome a signaling.

Phytochrome kinase substrate1 (PKS1) is a cytoplasmic protein that interacts physically with, and is phosphorylated by, the plant photoreceptor phytochrome. Here, we show that light transiently increases PKS1 mRNA levels and concentrates its expression to the elongation zone of the hypocotyl and root. This response is mediated by phytochrome A (phyA) acting in the very low fluence response (VLFR) mode. In the hypocotyl, PKS1 RNA and protein accumulation are maintained only under prolonged incubation in far-red light, the wavelength that most effectively activates phyA. Null mutants of PKS1 and its closest homolog, PKS2, show enhanced phyA-mediated VLFR. Notably, a pks1 pks2 double mutant has no phenotype, whereas overexpression of either PKS1 or PKS2 results in the same phenotype as the pks1 or pks2 single null mutant. We propose that PKS1 and PKS2 are involved in a growth regulatory loop that provides homeostasis to phyA signaling in the VLFR. In accordance with this idea, PKS1 effects are larger in the pks2 background (and vice versa). Moreover, the two proteins can interact with each other, and PKS2 negatively regulates PKS1 protein levels specifically under VLFR conditions.

LAF3, a novel factor required for normal phytochrome A signaling.

Phytochrome A (phyA) is the photolabile plant light receptor that mediates broad spectrum very low-fluence responses and high irradiance responses to continuous far-red light (FRc). An Arabidopsis mutant laf3-1 (long after far-red 3) was recovered from a screen for transposon-tagged mutants that exhibit reduced inhibition of hypocotyl elongation in FRc. The laf phenotype correlated well with a strongly attenuated disappearance of XTR7 transcript in FRc. The effects of laf3-1 on phyA-controlled CAB, CHS, and PET H expression were more subtle, and the mutation had no clear effects on PET E and ASN1 transcript levels in FRc. The use of two alternative transcription initiation sites in the LAF3 gene generates two isoforms that differ only at their N termini. Transcripts encoding both isoforms were induced during germination and were present at slightly higher levels in de-etiolated seedlings than in those grown in darkness. No significant differential regulation of the two isoforms was observed upon exposure to either FRc or continuous red light. Transcripts encoding the shorter isoform (LAF3ISF2) always appear to be more abundant than those encoding the longer isoform (LAF3ISF1). However, both isoforms were capable of full complementation of the laf3-1 hypocotyl phenotype in FRc. When fused to a yellow fluorescent protein, both isoforms localize to the perinuclear region, suggesting that LAF3 encodes a product that might regulate nucleo-cytoplasmic trafficking of an intermediate(s) involved in phyA signal transduction.

Light-mediated regulation defines a minimal promoter region of TOP2.

Light signaling has been demonstrated to be an important factor for plant growth and development; however, its role in the regulation of DNA replication and cell cycle has just started to be unraveled. In this work, we have demonstrated that the TOP2 promoter of Pisum sativum (pea) is activated by a broad spectrum of light including far-red light (FR), red light (RL) and blue light (BL). Deletion analyses of the TOP2 promoter in transformed plants, Arabidopsis thaliana and Nicotiana tobaccum (tobacco), define a minimal promoter region that is induced by RL, FR and BL, and is essential and sufficient for light-mediated activation. The minimal promoter of TOP2 follows the phytochrome- mediated low-fluence response similar to complex light regulated promoters. DNA-protein interaction studies reveal the presence of a DNA binding activity specific to a 106 bp region of the minimal promoter that is crucial for light-mediated activation. These results altogether indicate a direct involvement of light signaling in the regulation of expression of TOP2, one of the components of the DNA replication/cell cycle machinery.

Photomorphogenesis of rice seedlings: a mutant impaired in phytochrome-mediated inhibition of coleoptile growth.

A mutant showing a long coleoptile phenotype under white light was isolated from gamma-ray-mutagenized rice (cv. Nihonmasari). This mutant, named cpm1 (coleoptile photomorphogenesis 1), has been found to be impaired in phytochrome-mediated inhibition of coleoptile growth. Another outstanding feature of the mutant is impaired anthesis. Under red light (R), cpm1 coleoptiles elongate at a higher rate than wild-type (WT) coleoptiles, owing to substantially reduced responsiveness to R. This phenotype occurs in an age-dependent manner, and cpm1 coleoptiles become responsive to R as they elongate. The impairment was found in both very-low-fluence and low-fluence responses. Mutant coleoptiles also elongate longer than WT coleoptiles in darkness, but in this case the long coleoptile results from an extended elongation period. The cpm1 mutation does not affect the following phytochrome responses: the growth stimulation in submerged coleoptiles (uncovered in this study), potentiation of greening, and down-regulation of PHYA transcription. The cpm1 mutation does not significantly affect the level of spectroscopically detectable phytochrome and the transcription levels of three phytochrome genes (PHYA-C). It is concluded that the CPM1 gene is involved in the phytochrome signal transduction that specifically leads to growth inhibition. Some aspects of rice seedling photomorphogenesis are discussed in relation to the results obtained.

Multiple light inputs control phototaxis in Synechocystis sp. strain PCC6803.

The phototactic behavior of individual cells of the cyanobacterium Synechocystis sp. strain PCC6803 was studied with a glass slide-based phototaxis assay. Data from fluence rate-response curves and action spectra suggested that there were at least two light input pathways regulating phototaxis. We observed that positive phototaxis in wild-type cells was a low fluence response, with peak spectral sensitivity at 645 and 704 nm. This red-light-induced phototaxis was inhibited or photoreversible by infrared light (760 nm). Previous work demonstrated that a taxD1 mutant (Cyanobase accession no. sll0041; also called pisJ1) lacked positive but maintained negative phototaxis. Therefore, the TaxD1 protein, which has domains that are similar to sequences found in both bacteriophytochrome and the methyl-accepting chemoreceptor protein, is likely to be the photoreceptor that mediates positive phototaxis. Wild-type cells exhibited negative phototaxis under high-intensity broad-spectrum light. This phenomenon is predominantly blue light responsive, with a maximum sensitivity at approximately 470 nm. A weakly negative phototactic response was also observed in the spectral region between 600 and 700 nm. A deltataxD1 mutant, which exhibits negative phototaxis even under low-fluence light, has a similar action maximum in the blue region of the spectrum, with minor peaks from green to infrared (500 to 740 nm). These results suggest that while positive phototaxis is controlled by the red light photoreceptor TaxD1, negative phototaxis in Synechocystis sp. strain PCC6803 is mediated by one or more (as yet) unidentified blue light photoreceptors.

Brassinosteroid Mutants Uncover Fine Tuning of Phytochrome Signaling

Phytochromes (phy) A and B provide higher plants the ability to perceive divergent light signals. phyB mediates red/far-red light reversible, low fluence responses (LFR). phyA mediates both very-low-fluence responses (VLFR), which saturate with single or infrequent light pulses of very low fluence, and high irradiance responses (HIR), which require sustained activation with far-red light. We investigated whether VLFR, LFR, and HIR are genetically coregulated. The Arabidopsis enhanced very-low-fluence response1 mutant, obtained in a novel screening under hourly far-red light pulses, showed enhanced VLFR of hypocotyl growth inhibition, cotyledon unfolding, blocking of greening, and anthocyanin synthesis. However, eve1 showed reduced LFR and HIR. eve1 was found allelic to the brassinosteroid biosynthesis mutant dim/dwarf1. The analysis of both the brassinosteroid mutant det2 in the Columbia background (where VLFR are repressed) and the phyA eve1 double mutant indicates that the negative effect of brassinosteroid mutations on LFR requires phyA signaling in the VLFR mode but not the expression of the VLFR. Under sunlight, hypocotyl growth of eve1 showed little difference with the wild type but failed to respond to canopy shadelight. We propose that the opposite regulation of VLFR versus LFR and HIR could be part of a context-dependent mechanism of adjustment of sensitivity to light signals.

Brassinosteroid Mutants Uncover Fine Tuning of Phytochrome Signaling

Phytochromes (phy) A and B provide higher plants the ability to perceive divergent light signals. phyB mediates red/far-red light reversible, low fluence responses (LFR). phyA mediates both very-low-fluence responses (VLFR), which saturate with single or infrequent light pulses of very low fluence, and high irradiance responses (HIR), which require sustained activation with far-red light. We investigated whether VLFR, LFR, and HIR are genetically coregulated. The Arabidopsis enhanced very-low-fluence response1mutant, obtained in a novel screening under hourly far-red light pulses, showed enhanced VLFR of hypocotyl growth inhibition, cotyledon unfolding, blocking of greening, and anthocyanin synthesis. However,eve1 showed reduced LFR and HIR. eve1 was found allelic to the brassinosteroid biosynthesis mutantdim/dwarf1. The analysis of both the brassinosteroid mutant det2 in the Columbia background (where VLFR are repressed) and the phyA eve1 double mutant indicates that the negative effect of brassinosteroid mutations on LFR requires phyA signaling in the VLFR mode but not the expression of the VLFR. Under sunlight, hypocotyl growth of eve1 showed little difference with the wild type but failed to respond to canopy shadelight. We propose that the opposite regulation of VLFR versus LFR and HIR could be part of a context-dependent mechanism of adjustment of sensitivity to light signals.

Temporal and spatial pattern of light-dependent acquisition of competence for shoot formation in tomato hypocotyl. I Light pulse conditions

In the present paper, we compare, in photomorphogenic mutants of tomato, the effect of a single exposure to light of etiolated seedlings on acquisition of competence for subsequent adventitious shoot formation in hypocotyl segments cultured in vitro under continuous white it light. The results indicate that the efficiency of 1 hr white light is maximal and constant in time in apical segments, while decreases with age in middle and basal segments. In apical segments of wild type (WT), au and hp-1 mutant of tomato the promoting effect of 1 hr pulse of far red light was similar to that of red and blue light, while it was substantial but reduced in phyA,phyB1, and phyA/phyB1 mutants. The residual response displayed by phyA,phyB1 and phyA/phyB1 background indicates that besides phyA and phyBl some other phytochromes are involved in the far-red mediated response. Pre-irradiation with fluence of red light inducing very low fluence responses (VLFR, 10 - 8 - 10 - 6 mol m 2 ) as well as low fluence responses (LFR, > 10 - 6 mol m 2 ) enhanced the frequency of shoot formation in apical segments of all the genotypes, whereas it was ineffective in middle and basal segments of phyA, phyB1 and phyA/phyB1 mutants. Only in WT tomato VLFR induced substantial enhancement of capability for shoot regeneration in middle and basal segments. Thus presence of phyB1 and phyA appears to be absolutely necessary for LFR- and VLFR-induced acquisition of competence for shoot formation in middle and basal segments of tomato hypocotyl, whereas in apical segments the remaining phytochromes, besides phyA and phyBl, are sufficient.

Phytochrome E Controls Light-Induced Germination of Arabidopsis

Germination of Arabidopsis seeds is light dependent and under phytochrome control. Previously, phytochromes A and B and at least one additional, unspecified phytochrome were shown to be involved in this process. Here, we used a set of photoreceptor mutants to test whether phytochrome D and/or phytochrome E can control germination of Arabidopsis. The results show that only phytochromes B and E, but not phytochrome D, participate directly in red/far-red light (FR)-reversible germination. Unlike phytochromes B and D, phytochrome E did not inhibit phytochrome A-mediated germination. Surprisingly, phytochrome E was required for germination of Arabidopsis seeds in continuous FR. However, inhibition of hypocotyl elongation by FR, induction of cotyledon unfolding, and induction of agravitropic growth were not affected by loss of phytochrome E. Therefore, phytochrome E is not required per se for phytochrome A-mediated very low fluence responses and the high irradiance response. Immunoblotting revealed that the need of phytochrome E for germination in FR was not caused by altered phytochrome A levels. These results uncover a novel role of phytochrome E in plant development and demonstrate the considerable functional diversification of the closely related phytochromes B, D, and E.

Association of alpha-amylase and the R1 protein with starch granules precedes the initiation of net starch degradation in turions of Spirodela polyrhiza.

In turions of Spirodela polyrhiza (L.) Schleiden, net degradation of storage starch is controlled by a special low fluence response of phytochrome requiring illumination for several days. This light effect has been used to study protein-starch interactions that occur prior to and during net degradation of starch. Following various pretreatments on S. polyrhiza turions, native starch granules were isolated and two fractions of starch-related proteins were distinguished: proteins enclosed within the starch particles (starch-internalized proteins) and those attached to the surface (starch-associated proteins). The pattern of starch-associated proteins as resolved by SDS-PAGE was more complex than that of starch-internalized proteins and varied depending upon the pretreatment of the turions. Two starch associated proteins were identified immunochemically as alpha-amylase (EC 3.2.1.1) and the R1 protein (Lorberth et al. (1998) Nature Biotechnology 16: 473-477). Dark-pretreatment of non-dormant turions does not induce starch net degradation. Under these conditions, alpha-amylase and R1 were bound to the surface of the starch granules. Continuous illumination with red light induces a rapid degradation of starch. Within the first 24 h of illumination the level of starch-associated alpha-amylase transiently increased and subsequently decreased rapidly. Similarly, the amount of the starch-associated R1 also decreased during illumination. The dissociation of both alpha-amylase and R1 from the starch granules preceded the decrease in starch content. However, binding of the two proteins to starch granules remained unchanged when the turions did not perform net starch degradation (as observed during continuous darkness, orthophosphate deficiency, or dormancy of the turions). Thus, during net starch degradation, so far unidentified changes are postulated to occur at the surface of the starch particles that are relevant for protein binding. This conclusion was supported by in vitro studies in which the binding of purified beta-amylase (EC 3.2.1.2) to starch granules isolated from turions following various pretreatments was monitored. The enzyme did bind to starch granules prepared from dark-stored turions (in which starch degradation had not been initiated), but not to those isolated from illuminated (starch degrading) turions.

Light Regulation of Nitrate Reductase in Higher Plants: Which Photoreceptors are Involved?

Abstract: The regulation by light of nitrate reductase (NR) in higher plants is very complex. NR is firstly regulated by transcriptional control. In etiolated plants, phytochrome is the main photoreceptor involved and its low fluence response (LFR) is the common response mode. The effect of the very low fluence response (VLFR) has been reported for one of the isoforms in Arabidopsis thaliana, NIA2. The existence of a far‐red light‐mediated high irradiance response (HIR) has been demonstrated in phytochrome A over‐expressing tobacco, confirming classical photophysiological experiments. For Lemna aequinoctialis, the existence of a red light‐mediated high irradiance response has been assumed. In green plants, the effect of phytochrome on the level of NR protein is very modest, and the effect of light is mediated via photosynthesis, most probably through biochemical signals. The molecular basis of this switch from the action of phytochromes in etiolated tissue to effects of photosynthesis in green tissue is not known.The influence of light on NR is also regulated by fast post‐translational modification. This mechanism is based on the phosphorylation/dephosphorylation of a serine residue in the hinge 1 region of NR and the subsequent Mg2+/polyamine‐dependent binding of the phosphorylated form to a 14‐3‐3 protein. Several kinases phosphorylate NR, and phosphatases not only act on NR but also dephosphorylate NR kinases. No influence of phytochrome is detectable. Instead, post‐translational modification depends on the effect of photosynthesis. Photosynthesis may exert its effect on this process via biochemical products of photosynthesis (glucose, sugar phosphates), via substrates (nitrate, NADH) and/or by influencing Ca2+ flux.

Phytochrome and temperature control of seed germination in Muntingia calabura L. (Elaeocarpaceae)

The temperature range of germination of seeds of Muntingia calabura L. was 15-35° C and the optimum temperature at 35° C where more seeds germinated at short period. The fluence response curves demonstrated the involvement of phytochrome in the control of germination through the low fluence response. The effect of photoperiod showed that the species need long daily exposures to white light and that shade light inhibited completely seed germination, confirming as pioneer species that colonized only large gaps in the forest.

Phytochrome A-mediated inhibition of seed germination in tomato.

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.