The Optimum Angle of Geotropic Stimulation and its Relation to the Starch Statolith Hypothesis

Roots which are turned from their normal direction to directions at various angles with the plumb line develop the largest geotropic curvatures during a subsequent klinostat rotation period when the stimulation angle is well above the horizontal. In experiments with roots of Lepidium sativum L., the optimum is located at 120 to 140° when the stimulation time is between 2 and 15 min. If this fact is to be explained by the movements of amyloplasts in the root cap cells, one would expect roots which bad been kept inverted before the stimulation (so that the moveable amyloplasts are accumulated in the opposite end of the cells) to show an optimum angle well below 90°. — Pre-inversion of the roots did suppress the curvatures produced by stimulation at angles larger than 90° when measured after 10 to 30 min of klinostat rotation. This suppression may be taken as a support for the starch statolith hypothesis, since the amyloplasts in pre-inverted roots placed at angles exceeding 90° have a restricted opportunity to slide along the cell walls compared to non-inverted roots placed at the same angles. In pre-inverted roots measured after a period of klinostat rotation, however, no optimum was found at angles below 90°. When the stimulation time was 3.75 min, the response curves were nearly symmetrical about 90°. Stimulation for 15 min, on the other hand, resulted in curvatures which were much larger (although suppressed in comparison with non-inverted roots) when the stimulation angle was 165° than when it was 15°. During the 15 min stimulation period itself, however, pre-inverted roots curved 0.3° when stimulated at 15, but only 3.4° at 165°. This small difference was very highly significant and is in agreement with the starch statolith hypothesis insofar as the amyloplasts in pre-inverted roots placed at 15° have the greatest opportunity to slide along the cell walls. The lack of further development (and the actual decrease) of their curvatures during the subsequent klinostat rotation must then be due to other, depressing, factors, summarily designated as tonic. At angles above 90°, the tonic factors are either absent or even enhancing. Tbe tonic effects cannot be explained by amyloplast movements.     

Elimination of Geotropic Responsiveness in Roots of Cress (Lepidium sativum) by Removal of Statolith Starch

Roots of garden cress (Lepidium sativum L.) seedlings were made starch-free by treatment with gibberellic acid and kinetic for 29 hours at 35°C in the dark. After 3 hours of temperature adaptation at 21°C the starch-depleted roots were unable to respond to gravity, but elongated 0.48 mm por hour. Under the same conditions control roots pretreated in plain water at 21 and at 35°C elongated 0.64 and 0.33 mm per hour, respcetively (at 21°C). When the hormone-treated seedlings were illuminated, their roots reformed starch after 20 to 24 hours; simultaneously the geotropic responsiveness was restored. The results are interpreted in support of the statolith theory.     

Geotropic Response of Wheat Coleoptiles in Absence of Amyloplast Starch

Young coleoptiles of wheat (Triticum durum var. Henry), depleted of amyloplast starch by incubation at 30°C with gibberellin plus kinetin, retained their geotropic responsiveness. Depleted coleoptiles curved upward more slowly than controls, but this was commensurate with their slower growth. The ratio of curvature to growth was about 50° per mm of elongation in both cases. Newly excised coleoptiles, though containing much more starch than incubated controls, curved only about 25° per mm. The tissue treated in gibberellin plus kinetin appeared to contain no starch when examined (a) freshly squashed, (b) as fixed material sectioned thin and stained by the PAS procedure, and (c) as electron micrographs. Shrunken, starch-free amyloplasts could be identified in certain regions, but these did not show evidence of asymmetric distribution under the influence of gravity. The possibilities that other organelles function as statoliths are considered, and it is concluded not only that georeception is independent of starch grains, but further that it may not be due to statoliths at all in an ordinary sense.     

Investigations of the Geotropic Curvature of the Avena Coleoptile

The geotropic reaction in Avena coleoptiles is studied as a function of the stimulation time. The direction of the stimulation with respect to the vascular bundles must be defined when studying geotropic responses. It is found that the threshold time to evoke geotropic response is less than half a minute, i.e., at least ten times lower than the presentation time usually reported in the literature. An extrapolation procedure can be used to give a so-called extrapolated presentation time t_b, which is intimately related to the logarithmic part of the geotropic response curve and has a physical meaning in the reciprocity rule. The problem of the duration of the true threshold time for stimulation with 1 g is discussed. An experiment indicates that it is not necessary for mass particles (“statoliths”) to settle on the lateral cell wall in order to start the geotropic reaction chain. The slope of the logarithmic part of the geotropic response curve is independent of the transverse force applied to the coleoptiles. Support is given to the view that the slope is determined by the number of sedimenting mass particles.     

Orientation and Geotropic Reaction of Seminal Roots of Wheat

The geotropic orientation of seminal roots of wheat has been studied on seedlings grown in five different positions, stationary and on clinostats. The roots perceive a geoinduc-tion before they have emerged from the grain and perform curvatures inside the grain. These are very sharp and transient, the following root growth is straight in any direction unless the positions are shifted. The roots are insensitive to a static gravi-induction but react to a change in gravitation with a geotropic curvature in positive direction. The roots may not reach or reach, or even pass the plumb-line. The orientation of a root depends upon the direction of its initiation and the geotropic curvature attained before the reaction has ceased. There is no nastic component in the reactions. The ‘plagiotropic’ orientation is explained by the limited positive reaction followed by an ageotropic state. Main root and adventitious roots react in the same way. Reactions to later stimuli give likewise limited curvatures which are weaker but of longer duration. — The effect of temperatures from 10°C to 25°C has been studied and compared to the temperature effect on cell elongation. It is concluded that the whole reaction may be explained by the regular auxin effects on cell elongation. No other hormone should be required and no plagiotropic mechanism is necessary.     

Diurnal Geotropic responses of the Primary Leaves of Phaseolus vulgaris

When seedlings of Phaseolus vulgaris with leaves in the daytimeposition (almost horizontal to the ground) were turned upside-downduring the light period, their leaves moved upward away fromthe ground after about 20 min and ceased moving after about1.5 h. But when seedlings with leaves in the night time position(directed downward) were turned upside-down, their leaves moveddownward toward the ground after about 30 min and stopped movingabout 2 h later. Thus, Phaseolus primary leaves showed positiveor negative geotropic responses that correspohded to the darkor light period. This geotropic response of primary leaves was accompanied bythe redistribution of K⁺, Cl^– and NO₃- in the laminarpulvinus. These facts suggest that the circadian endogenousclock that is assumed to exist in Phaseolus vulgaris has atleast two regulation echanisms; one which measures time andanother which determines leaf postition in relation to gravityby changing the ion distribution in the pulvinus (Received February 12, 1983; Accepted May 17, 1983)     

The Effects of Paraffin Oil on Phototropic and Geotropic Responses in Avena Coleoptiles

Experiments are described which were designed to test the significanceof the coleoptile tip as the site of reception of light stimulusleading to negative photo-tropic response under paraffin oil.The results show clearly that the tip is of paramount importancein this respect. Further experiments in which the coleoptiletips were bisected in a plane at right angles to the light rayslend support to the hypothesis that negative phototropism underoil is related tolateral transport of material in the tip. Lastly,experiments which show that the geotropic response of coleoptilesis not reversed by immersion in oil are described. These findingsare discussed in relation to certain hypotheses concerning themechanism of negative phototropism under oil.     

Effects of Exogenous Thyroxine on Statolith Synthesis and Resorption in Aurelia

Aurelia ephyrae which were maintained in thyroxine during theirmetamorphosis from the polyp state and another group of ephyraewhich were in thyroxine forfour and eight days of starvation,had significantly reduced numbers of statoliths as comparedwith control ephyrae in equimolar concentrations of iodine andin artificial sea water. We conclude, therefore, that thyroxinesignificantly affects the mineralization and demineralizationof statocytes resulting in changes in statolith numbers. A mineralization role of thyroxine in other invertebrates hasnot been reported in spite of the fact that thyroxine and itsprecursors, monoiodotyrosine and diiodotyrosine, have been locatedin numerous invertebrates, particularly in mineralizing areassuch as developing shells. On the other hand, an important roleof thyroxine in mineralization and demineralization of bonesand teeth of vertebrates including humans, has been known forsome time. The specific role of thyroxine in affecting mineralizationin vertebrates is not known, due perhaps to the presence ofmany other hormones and second messengers which also affectmineralization. The use of the relatively simple Aurelia statolithtest systems and the exploration for other invertebrate mineralizingsystems which respond to thyroxine will hopefully lead to abetter understanding of the basic mechanisms of thyroxine actionin mineralizing systems in the future.