Phytosulfokine-alpha, a peptide growth factor found in higher plants: its structure, functions, precursor and receptors

Phytosulfokine-alpha, a sulfated pentapeptide growth factor universally found in both monocotyledons and dicotyledons, strongly promotes proliferation of plant cells in culture. It is similar to animal polypeptide hormones in that it is processed from a larger precursor, preprophytosulfokine, although the putative processing sites do not conform to consensus sequences for endoproteolytic processing sites flanking animal prohormones. Like the animal preprohormones, preprophytosulfokine also has a signal peptide at the N-terminus for targeting to secretory pathways. The preprophytosulfokine gene has been confirmed to be expressed in vivo as well as in vitro.     

A rapid and efficient system of Agrobacterium infection-mediated transient gene expression in rice Oc cells and its application for analysis of the expression and antisense suppression of preprophytosulfokine, a precursor of phytosulfokine-a, encoded by OsPSK gene

A rapid and efficient system for Agrobacterium infection-mediated transient gene expression in rice has been developed. Using this system, transient expression of preprophytosulfokine, a precursor of phytosulfokine-a, encoded by OsPSK gene was analyzed. The results suggest that the Agrobacterium infection-mediated transient gene expression system is as efficient in rice Oc cells as in tobacco BY-2 cells and might be useful for rapid analysis not only of foreign gene expression, but also of antisense gene suppression.     

A secreted peptide growth factor, phytosulfokine, acting as a stimulatory factor of carrot somatic embryo formation

Somatic embryogenesis of the carrot (Daucus carota L.) depends on a set of factors, some of which accumulate in culture medium (conditioned medium, CM). When embryogenic cell clusters were transferred to an embryo-inducing medium, addition of CM derived from somatic embryo culture markedly stimulated somatic embryo formation. The active principles were purified using a simple bioassay system and identified to be phytosulfokines (PSKs), sulfated oligopeptide growth factors originally isolated from a CM derived from asparagus (Asparagus officinalis L.) mesophyll culture. Quantification studies using a competition ELISA system employing an anti-PSK-alpha polyclonal antibody showed that PSK production might be related to growth of cells, rather than development of somatic embryos. Thus the stimulatory effect of PSK on somatic embryo formation might be due to promotion of cell proliferation.     

Phytosulphokine-{alpha}, a peptidyl plant growth factor, stimulates somatic embryogenesis in carrot

Phytosulphokine- (PSK-) is the first chemically characterized peptide that acts as a plant growth factor. It stimulates the proliferation of asparagus and rice cells, but no information is yet available on its effects on plant morphogenesis. The effects of PSK- on somatic embryogenesis in carrot (Daucus carota L.) were examined. PSK-, when added to the induction medium for somatic embryogenesis, increased the number of somatic embryos. The chemical analogues [2-5]PSK- and tyrosine sulphate ester (Tyr-SO3 H), which have been used as negative controls in other systems, had no effect. Moreover the proliferation of cells during somatic embryogenesis was also enhanced by PSK- these results indicate that PSK- enhanced cell division and, as a consequence, stimulated carrot somatic embryogenesis. PSK- also stimulated the proliferation of embryogenic cells in medium that contained 2,4-dichlorophenoxyacetic acid (2,4-D), in which somatic embryos did not form, as well as the proliferation of non-embryogenic cells (cells that had lost the ability to form somatic embryos) in medium without 2,4-D. These results indicate that PSK- has a stimulatory effect on cell division generally in carrot cell cultures.Key words: Daucus carota, plant growth factor, somatic embryogenesis, sulphated peptide.      

Promotive effects of the peptidyl plant growth factor, phytosulfokine-alpha, on the growth and chlorophyll content of Arabidopsis seedlings under high night-time temperature conditions

In order to investigate the function of the peptidyl plant growth factor, phytosulfokine-alpha (PSK-alpha), in plants, we examined the effect of PSK-alpha on the growth and chlorophyll content of Arabidopsis seedlings under high night-time temperature conditions. Although exposure to high night-time temperatures markedly reduced the fresh weight and chlorophyll content of the seedlings, these parameters in the plants supplied with PSK-alpha remained at the same levels as those of non-treated controls. These effects were not apparent when [2-5]PSK, Tyr-SO3H and kinetin were similarly supplied. The results suggest that PSK-alpha not only promotes cell proliferation, but may aid plants in their tolerance of heat stress.     

A video-sensor system for predicting the time of parturition in the chimpanzee

A system of predicting the time of parturition in the chimpanzee was developed to assure care of the newborn as soon as possible after delivery. To quantify activity, the number of times that a parturient chimpanzee crossed a sensor marker of a TV monitor was recorded by a videosensor on the market for crime prevention. The amount of activity increased remarkably just prior to parturition. A system using a personal pocket alarm ("beeper"), which rang automatically when the number of crossings by the chimpanzee exceeded a selected level within a certain period of time, was tested. In the present study, the threshold for sounding the alarm was set at 50 crossings within 10 minutes. As a result, the pocket alarm began to ring at about 40 minutes before parturition. This enabled us to be present at parturition and to tend to a newborn abandoned by its mother in one case. The present system could be useful to predict the time of parturition in the chimpanzee.     

Purification of chloroplast elongation factor Tu and cDNA analysis in tobacco: the existence of two chloroplast elongation factor Tu species

We have purified a chloroplast elongation factor Tu (EF-Tu) from tobacco (Nicotiana tabacum) and determined its N-terminal amino acid sequence. Two distinct cDNAs encoding EF-Tu were isolated from a leaf cDNA library of N. sylvestris (the female progenitor of N. tabacum) using an oligonucleotide probe based on the EF-Tu protein sequence. The cDNA sequence and genomic Southern analyses revealed that tobacco chloroplast EF-Tu is encoded by two distinct genes in the nuclear genome of N. sylvestris. We designated the corresponding gene products EF-Tu A and B. The mature polypeptides of EF-Tu A and B are 408 amino acids long and share 95.3% amino acid identity. They show 75–78% amino acid identity with cyanobacterial and chloroplast-encoded EF-Tu species.     

Asymmetry of membrane fluidity in the lipid bilayer of blood platelets: Fluorescence study with diphenylhexatriene and analogs

Summary Membrane fluidity of bovine platelets was examined with diphenylhexatriene (DPH), its cationic trimethylammonium derivative (TMA-DPH) and anionic propionic acid derivative (DPH-PA). After addition of these probes to platelet suspensions at 37°C, the fluorescence intensity of DPH-PA reached equilibrium within 2 min, whereas those of DPH and TMA-DPH increased gradually. With increase in the fluorescence intensity of TMA-DPH, its fluorescence anisotropy decreased significantly, but the fluorescence anisotropies of DPH-PA and DPH did not change during incubation. The gradual increase of fluorescence intensity of TMA-DPH was due to its penetration into the cytoplasmic side of the platelet membrane, as shown quantitatively by monitoring decrease in its extractability with albumin. Transbilayer movement of TMA-DPH was markedly temperature-dependent, and was scarcely observed at 15°C. The fluorescence intensity of TMA-DPH was much higher in platelet membranes and vesicles of extracted membrane lipids than the initial intensity in intact platelets. Moreover, the fluorescence anisotropy of TMA-DPH was much lower in the former preparations than the initial value in intact platelets. These results suggest that binding sites for TMA-DPH in the cytoplasmic side of the platelet membrane are more fluid than those in the outer leaflet of the plasma membrane. Platelet activation by ionomycin induced specific change in the fluorescence properties of TMA-DPH without causing transbilayer incorporation of the probe.