In vitro food production for isolated closed environments: formation of ripe tomato fruits from excised flower buds.

Excised preanthesis flower buds of young Pixie Hybrid tomato plants develop into red ripe fruits in aseptic culture on a modified Murashige-Skoog medium with 3% sucrose at pH 5.8. The addition of certain synthetic auxins (IAA, NAA, IBA), auxin precursors (ISA), or cytokinins (KIN, IPA, ZEA, BAP) to the medium improved the percentage of buds developing into fruits, the weight of the ripe fruits, or both. The best results were obtained by an auxin-cytokinin combination of 10 microM IBA with 1 microM BAP. Storage of the excised buds at low temperature (6 degrees C) for up to 4 weeks before transfer to 27 degrees C caused only minimal deterioration in size and number of the fruit crop. Extension of low-temperature storage to 8 weeks produced smaller fruits that took longer to develop. This system could produce fresh, ripe small tomatoes on a sustained basis for up to 2 months for an isolated environment such as a space vehicle or submarine.

Isatin as an auxin source favoring floral and vegetative shoot regeneration from calli produced by thin layer explants of tomato pedicel.

Thin layer explants taken from the pedicels and peduncles of flowering tomato plants yielded calli with great organogenetic potential. Of the 15 cultivars tested, 7 regenerated roots, shoots and eventually entire fruit-bearing plants. Calli grown on modified Murashige-Skoog medium responded to varied auxins and cytokinins with different morphogenetic patterns. Thus, naphthaleneacetic acid yielded root-producing calli, while the auxin precursor isatin (indole 2,3-dione) caused the production of calli with vegetative and floral shoots, rarely yielding roots. This may be related to isatin's slow, steady conversion to an active auxin (Plant Physiol 41:1485-1488, 1966) in contrast with naphthaleneacetic acid's immediate presentation of a high level of active auxin. The highest incidence of vegetative shoot (100%) and flower (50%) formation was obtained with 10 micromoles isatin and 3 micromoles zeatin. A few of the flowers developed into ripe fruits. The high frequency of induction of vegetative shoots and flowers before roots with isatin suggests its utility in micropropagation from plant tissue cultures.

Polyamine binding to proteins in oat and Petunia protoplasts.

Previous work (A Apelbaum et al. [1988] Plant Physiol 88: 996-998) has demonstrated binding of labeled spermidine (Spd) to a developmentally regulated 18 kilodalton protein in tobacco tissue cultures derived from thin surface layer explants. To assess the general importance of such Spd-protein complexes, we attempted bulk isolation from protoplasts of Petunia and oat (Avena sativa). In Petunia, as in tobacco, fed radioactive Spd is bound to protein, but in oat, Spd is first converted to 1,3,-diaminopropane (DAP), probably by polyamine oxidase action. In oat, binding of DAP to protein depends on age of donor leaf and conditions of illumination and temperature, and the extraction of the DAP-protein complex depends upon buffer and pH. The yield of the DAP-protein complex was maximized by extraction of frozen-thawed protoplasts with a pH 8.8 carbonate buffer containing SDS. Its molecular size, based on Sephacryl column fractionation of ammonium sulfate precipitated material, exceeded 45 kilodaltons. Bound Spd or DAP can be released from their complexes by the action of Pronase, but not DNAse, RNAse, or strong salt solutions, indicating covalent attachment to protein.

Binding of spermidine to a unique protein in thin-layer tobacco tissue culture.

The mechanism by which spermidine induces the appearances of floral buds in thin-layer tobacco (Nicotiana tabacum) tissue culture was studied by following the fate of the radioactive compound. [3H]Spermidine was taken up rapidly by the tissue, and after a brief lag, a portion was bound to trichloroacetic acid precipitable macromolecules. Such binding increased to a maximum on day 4 of culture, coinciding with the onset of bud differentiation, and declined thereafter until shortly before flowering. About 82% of the label in the trichloroacetic acid precipitate remained as spermidine, 14% was metabolized to putrescine, 3% to spermine, and 1% to gamma-aminobutyric acid. Spermidine was covalently bound to a protein with a molecular size of about 18 kilodaltons. Hydrolysis of this protein and analysis of the labeled entities revealed 81% spermidine, 16% putrescine, and 3% spermine. This post-translational modification of a unique protein by attachment of spermidine may be causally connected to the appearance of flower buds in thin-layer tobacco cultures.

A Simple and Automatic Leaflet Movement-monitoring System.

A simple and automatic leaflet movement-monitoring system using phototransistors is described for Albizzia julibrissin.

Protein synthesis inhibition alters Drosophila mating behavior.

Fruit fly Drosophila pseudoobscura virgin AR females mate preferentially with AR males given a choice between AR and or males. However, AR females which mated with or males when young show a significant change in sexual selection in favor of or over AR males in subsequent simultaneous choice tests. Ingestion of food moistened with 4 ml cycloheximide (400 microng/ml) produces 75% protein synthesis inhibition in the female flies. Females exposed to cycloheximide (CXM) immediately after their initial copulation with or males resemble virgin flies in their choice of mates, and mate preferentially with AR males. Females exposed to CXM before their initial copulatory experience with or males resemble or-experienced but untreated (no CXM) flies in their choice of mates, and mate preferentially with or males. The change in sexual preference shown by females with prior copulatory experience resembles learning in that it is subject to disruption by CXM in ways analogous to those reported in the literature.

Potassium flux and leaf movement in Samanea saman. I. Rhythmic movement.

Samanea leaflets usually open in white light and fold together when darkened, but also open and dose with a circadian rhythm during prolonged darkness. Leaflet movement results from differential changes in the turgor and shape of motor cells on opposite sides of the pulvinus; extensor cells expand during opening and shrink during closure, while flexor cells shrink during opening and expand during closure but change shape more than size. Potassium in both open and closed pulvini is about 0.4 N. Flame photometric and electron microprobe analyses reveal that rhythmic and light-regulated postassium flux is the basis for pulvinar turgor movements. Rhythmic potassium flux during darkness in motor cells in the extensor region involves alternating predominance of inwardly directed ion pumps and leakage outward through diffusion channels, each lasting ca 12 h. White light affects the system by activating outwardly directed K(+) pumps in motor cells in the flexor region.

Rhythmic potassium flux in albizzia: effect of aminophylline, cations, and inhibitors of respiration and protein synthesis.

Rhythmic leaflet movement in Albizzia is controlled by rhythmic K(+) flux in pulvinal motor cells. The angle assumed by darkened leaflets during the open phase of the rhythm can be altered by various compounds and changes in temperature; such treatments are ineffective during the closed phase. In all cases, effects on leaflet angle are correlated with and probably a consequence of K(+) flux in pulvinal motor cells. Incubation at low temperature (6C) or on sodium azide (1.0 mm) reduces K(+) in the ventral motor region and increases K(+) in the dorsal motor region, thereby decreasing leaflet angle. Incubation on cycloheximide (0.1 mm) or sodium acetate (0.05 m) inhibits protein synthesis; if the incubation period immediately precedes the opening phase, these compounds prevent both K(+) flux into the ventral motor cells and leaflet opening. Magnesium nitrate (0.05 m), supplied after leaflets have started to open, promotes K(+) secretion from the dorsal motor cells and increases the angle of opening.The data support the following hypothesis. Active K(+) transport into the ventral motor cells and out of the dorsal motor cells leads to opening; K(+) leakage in the opposite directions causes closure; and the interaction of these processes results in a rhythmic oscillation. Proteins in the ventral cell membranes that are required for active transport or membrane integrity turn over rhythmically and are resynthesized before opening. The availability of divalent cations determines the phase relationships between the K(+) rhythms in the dorsal and ventral motor cells.White light phases the rhythm. The "light on" signal turns on a K(+) secreting pump in the dorsal motor cells, while the "light off" signal initiates a period of protein synthesis which in turn leads to active K(+) transport into the ventral motor cells. Aminophylline (0.1-6.0 mm) inhibits white light-promoted opening and nyctinastic closure.

Protein synthesis during endogenous rhythmic leaflet movement in Albizzia.

A rhythm was found in protein synthesis accompanying rhythmic leaflet opening and closing in the dark in the plant Albizzia. More protein was synthesized during the opening of the leaflets than during the closing. Furthermore, an inhibitor of protein synthesis, cycloheximide, prevented rhythmic opening of leaflets but had no effect on rhythmic closing. It is suggested that protein synthesis is involved in the movement across membranes of K(+) ions that cause turgor changes and leaflet movement.

Rhythmic Leaflet Movement in Albizzia julibrissin: Effect of Electrolytes and Temperature Alteration.

The rhythmic movement of darkened Albizzia leaflets is accompanied by K(+) flux in pulvinule motor cells whose turgor changes control opening and closing. The azide-sensitive open phase is promoted by an increase in temperature from 16 to 33C (Q(10) = 3), implying active transport of K(+) ions during this period. The azide-insensitive closed phase is less temperature-sensitive and has a Q(10) less than 1, implying diffusion or some other physical process as the predominant pathway of K(+) flux at this time. Thus rhythmic leaflet movement is probably due to oscillation in active K(+) transport or membrane permeability or both. External electrolytes (0. 1 n) alter leaflet angle during the open, but not the closed, phase of the rhythm. All chlorides except NH(4) (+) promote opening, with divalent more effective than monovalent ions. Some anions promote and others inhibit opening; activity is not correlated with charge. It is likely that electrolytes alter leaflet movement by altering K(+) flux, accomplishing this by interacting with key macromolecules in motor cell membranes.Pfr phytochrome dampens the amplitude of rhythmic leaflet movement; this process is temperature sensitive (Q(10) = 2) and unaltered by 0.1 n salt solutions. Although K(+) flux is a common basis for phytochrome and rhythmic control of leaflet movement, different mechanisms are clearly involved.