Phytochrome

In the most comprehensive study to date we explored the phylogeny and evolution of the genus Asparagus, with emphasis on the southern African species. We included 211 accessions, representing 77 (92%) of the southern African, 6 (17%) of the tropical African, 10 (56%) of the strictly European and 6 (9%) of the Eurasian species. We analysed DNA sequences from three plastid regions (trnH-psbA, trnD-T, ndhF) and from the nuclear region phytochrome C (PHYC) with parsimony and maximum likelihood methods, and recovered a monophyletic Asparagus. The phylogeny conflicts with all previous infra-generic classifications. It has many strongly supported clades, corroborated by morphological characters, which may provide a basis for a revised taxonomy. Additionally, the phylogeny indicates that many of the current species delimitations are problematic. Using biogeographic analyses that account for phylogenetic uncertainty (S-DIVA) and take into account relative branch lengths (Lagrange) we confirm...

Acetylcholine (ACh) has been detected in representatives of many taxonomic groups throughout the plant kingdom. The site of its synthesis in plants is probably young leaves. In some plant species choline acetyltransferase (ChAT) activity has been found. This enzyme showing properties similar to animal ChAT, probably participates in ACh synthesis from its precursors, choline and acetyl-Coenzyme A. Acetylcholinesterase (AChE) activity has also been found in many plant tissues. This enzyme decomposes ACh and exhibits properties similar to animal AChE. The presence of both ChAT and AChE in plant tissues suggests that ACh undergoes similar metabolism in plants as it does in animals. Exogenous ACh affects phytochrome-controlled plant growth and development. Mimicking red light (R), ACh stimulates adhesion of root tips to a glass surface and influences leaf movement and membrane permeability to ions. It also affects seed germination and plant growth. Moreover, ACh can modify some enzyme activity and the course of some metabolic processes in plants. Acetylcholine in the presence of calcium ions (Ca2+), like R stimulates swelling of protoplast isolated from etiolated wheat leaves. It is proposed that the primary mechanism of action of ACh in plant cells is via the regulation of membrane permeability to protons (H+), potassium ions (K+), sodium ions (Na+) and Ca2+. Acetylcholin (ACh) wurde in Vertretern vieler taxonomischer Gruppen des Pflanzreiches gefunden. Es wird wahrscheinlich inden jungen Blättern synthetisiert. In einigen Pflanzen hat man daneben Cholin-Acetyltransferase (ChAT)-Activität nachweisen können; dieses Enzym ziegt ähnliche Eigenschaften wie tierische ChAT und ist offenbar an der ACh-Synthese aus sienen Vorstufen Cholin und Acetyl-Coenzym A beteiligt. Acetylcholineesterase (AChE)-Activität wurde ebenfalls in vielen Pflanzengeweben gefunden; dieses Enzym spaltet ACh und ziegt ähnliche Eigenschaften wie tierische AChE. Die Anwesenheit von ChAT und AChE in pflanzlichem Gewebe läßt vermuten, daß ACh in Pflanzen einem ähnlichen Metabolismus unterliegt wie im tierischen System. Ähnlich wie Rotlicht stimuliert ACh die Anheftung von Wurzelspitzen an Glasoberflächen und beeinflußt Blattbewegung und Membranpermeabilität für Ionen; darüber hinaus beeinflußt es Samenkeimung und pflanzliches Wachstum. Des weiteren kann ACh Enzym-Aktivitäten modifizieren und dadurch den Ablauf einiher metabolischer Prozesse in Pflanzen. Schließlich stimuliert ACh in Gegenwart von Calcium-Ionen (Ca2+), ähnlich wie Rotlicht, das Schwellen von Protoplasten etiolierter Weizenblätter. Es wird vermutet, daß die Primärwirkung von ACh in Pflanzenzellen durch Regulation der Membranpermeabilität für Protonen (H+), Kaliumionen (K+), Natriumionen (Na+) und Ca2+ erfolgt.

PHYTOCHROME-INTERACTING FACTOR5 (PIF5), a basic helix-loop-helix transcription factor, interacts specifically with the photoactivated form of phytochrome B (phyB). Here, we report that dark-grown Arabidopsis thaliana seedlings overexpressing PIF5 (PIF5-OX) exhibit exaggerated apical hooks and short hypocotyls, reminiscent of the triple response induced by elevated ethylene levels, whereas pif5 mutants fail to maintain tight hooks like those of wild-type seedlings. Silver ions, an ethylene receptor blocker, rescued the triple-response phenotype, and we show that PIF5-OX seedlings express enhanced levels of key ethylene biosynthesis enzymes and produce elevated ethylene levels. Exposure of PIF5-OX seedlings to prolonged continuous red light (Rc) promotes hypocotyl elongation relative to dark controls, the reciprocal of the Rc-imposed hypocotyl inhibition displayed by wild-type seedlings. In contrast with this PIF5-OX hyposensitivity to Rc, pif5 mutant seedlings are hypersensitive rela...

Life occurs in an ever-changing environment. Some of the most striking and predictable changes are the daily rhythms of light and temperature. To cope with these rhythmic changes, plants use an endogenous circadian clock to adjust their growth and physiology to anticipate daily environmental changes. Most studies of circadian functions in plants have been performed under continuous conditions. However, in the natural environment, diurnal outputs result from complex interactions of endogenous circadian rhythms and external cues. Accumulated studies using the hypocotyl as a model for plant growth have shown that both light signalling and circadian clock mutants have growth defects, suggesting strong interactions between hypocotyl elongation, light signalling and the circadian clock. Here, we review evidence suggesting that light, plant hormones and the circadian clock all interact to control diurnal patterns of plant growth.

The light-regulated mRNA and polypeptide accumulation of the nuclear encoded subunit II (PsaD) of the photosystem I reaction center was studied during the greening of etiolated spinach seedlings. Upon exposure to continuous white light, the mRNA, detected at low levels in etiolated seedlings, accumulated in a specific pattern. In contrast, the PsaD subunit could not be detected in the etiolated seedlings; the polypeptide could first be detected in thylakoid membranes approximately 4 h after exposure to continuous light. A pulse of red light induced the expression of the PsaD mRNA, but the polypeptide could not be detected unless the seedlings were exposed to light. In the light (but not in the dark), the PsaD mRNA was found associated with the polysomal fraction. Taken together, the data suggest a dual regulatory mechanism in which both the level of mRNA and the presence of light control the accumulation of the PsaD polypeptide.

Plants use phytochrome (phy) photoreceptors to detect and respond to changes in the quantities and proportions of red (R) and far-red (FR) light in their environments. The principal mediators of responses to R and FR in Arabidopsis thaliana are phyA and phyB, which are found in all angiosperms surveyed. The present study is concerned with a phytochrome gene pair in Arabidopsis, PHYB and PHYD, which are of relatively recent origin, share high sequence identity, and are partially redundant. Our data suggest that the duplication occurred after the mustard family (Brassicaceae) diverged from its closest relatives but before the radiation of extant Brassicaceae, and that both copies have persisted for up to 40 myr. We detected no evidence of positive selection in the divergence of PHYD from PHYB; the evolution of both sequences is constrained by purifying selection. Levels of diversity at both loci are among the lowest observed at nuclear genes in A. thaliana. In common with other loci in A. thaliana, PHYB and PHYD showed elevated levels of intraspecific replacement variation, and each showed an excess of rare nucleotide polymorphisms, consistent with a recent, rapid population expansion. Our results are consistent with the functional importance of amino acid divergence in the central regions of phyB and phyD and suggest specific sites for mutagenesis that may yield insights into the functional differences of phyB and phyD.

The structure of the gene encoding the apoprotein of phytochrome B1 (PHYB1) in tomato has been determined from genomic and cDNA sequences. In contrast to PHYA, PHYB1 lacks an intron upstream of the first ATG. A single transcription start site was found by 5′ RACE at –116. Tomato PHYB1 spans 7 kb starting from the first ATG. The coding region is organized into four exons as for other angiosperm PHY. The deduced apoprotein consists of 1131 amino acids, with a molecular mass of 125.4 kDa. Tomato phytochrome B1 shares 78% and 74% identity with Arabidopsis phytochromes B and D, respectively. Along with the normally spliced full-length transcripts, sequences of reverse transcriptase-PCR clones revealed five types of alternative transcripts. Each type of alternative transcript was missing a considerable part of the coding region, including the chromophore-binding site. The four putative PHYB1 mutants in tomato, which are temporarily red-light insensitive (tri), were each confirmed to have a mutation in PHYB1. Each mutation arose from a different, single-base substitution. Allele tri1 is presumably a null because the mutation introduces a stop at codon 92. In tri3, val-238 is replaced by Phe. The importance of this valine residue is evidenced by the fact that the tri3 phenotype is as strong as that of tri1. Alleles tri2 and tri4 encode proteins truncated at their C-termini. The former lacks either 170 or 438 amino acids, depending upon which of two types of splicing occurs during transcript maturation, while the latter lacks 225.

Using sucrose gradient centrifugation, anion exchange chromatography on a SMART system, isoelectric focusing (IEF) and sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS PAGE), a 45.5 kDa membrane protein was isolated from amoebae of the cellular slime mould Dictyostelium discoideum. The absorption spectrum of the isolated protein corresponds well with the action spectrum for the photoaccumulation of the amoebae of Dictyostelium discoideum showing a major peak between 405 and 412 nm and some minor peaks between 500 and 600 nm. The remarkable similarity of the two spectra leads to the hypothesis that the isolated protein-pigment complex may serve as a photoreceptor for amoebal phototaxis.

Rapid responses to changes in incident light are critical to the guidance of behavior and development in most species. Phytochrome light receptors in particular play key roles in bacterial physiology and plant development, but their functions and regulation are less well understood in fungi. Nevertheless, genome-wide expression measurements provide key information that can guide experiments that reveal how genes respond to environmental signals and clarify their role in development. We performed functional genomic and phenotypic analyses of the two phytochromes in Neurospora crassa, a fungal model adapted to a postfire environment that experiences dramatically variable light conditions. Expression of phy-1 and phy-2 was low in early sexual development and in the case of phy-2 increased in late sexual development. Under light stimulation, strains with the phytochromes deleted exhibited increased expression of sexual development-related genes. Moreover, under red light, the phy-2 knoc...