Production of ethylene by gravistimulation: a potential problem with the interpretation of data from some experimental techniques

Abstract. Gravistimulation was investigated as a potential and unwanted component in the interpretation of physiological investigations on plants. Using both seedlings and mature sunflower plants, two situations are described where gravistimulation contributes significantly to the outcome of an experiment not initially designed to include this parameter. The number of adventitious roots formed in derooted seedings decreased when the tops of the plants were allowed to bend over under their own weight, and the effect correlated positively with the rate of ethylene production by non-vertical stems. In droughted mature plants, and increase in leaf and stem ethylene caused by water stress was supplemented by additional ethylene produced in the lower halves of stems. Drought had caused these stem tissues to wilt, become gravistimulated, and thus produce more ethylene. Other situations in which gravistimulated ethylene production and its physiological consequences are likely to be complicating factors in experiments are discussed. It is concluded that procedures that unnecessarily place experimental material in non-vertical orientations should be avoided.     

Immunochemical characterization and localization of nitrate reductase in norflurazon-treated soybean cotyledons

Immunochemical procedures were used to characterize and localize NADH:nitrate reductase (NR; EC 1.6.6.1) in cotyledons of norflurazon-treated soybeans [ Glycine max (L.) Merr. cv. 'Hill']. Antiserum prepared to NR isolated from Chlorella strongly reacted against NR from norflurazon-treated cotyledons. This serum inhibited the NR activity in crude extracts of norflurazon-treated soybean cotyledons by 98% even at a 1:2000 dilution of crude serum. Pre-immune serum had no effect on the activity. These data indicate that there are similar antigenic determinants at the active site of both Chlorella and norflurazon-treated soybean NR. Whole cotyledons were homogenized in lithium dodecyl sulfate-containing buffer, electrophoretically separated and blotted to nitrocellulose. When the blots were reacted with the anti-NR serum only a single protein (M_r= 98 kdalton) was visualized. Immunofluorescence studies on fixed tissue sections revealed intense fluorescence in the cytoplasm. Weaker reactions were associated with organelles tentatively identified as plastids. Pre-immune serum controls were completely unstained using immunocytochemical procedures.     

Relation of blood-free to blood-inclusive postmortem lung water measurements in sheep

Our purpose was to see if the postmortem weight ratio of extravascular lung water to blood-free dry lung (blood-free ratio) was related to similar ratios in blood-inclusive lung and in blood. We developed linear regressions of blood-free ratio on ratios for blood-inclusive lung and blood together and for blood-inclusive lung alone for 73 sheep studied under 11 different protocols and for two subgroups of sheep, one with plasma space expansion and the other without expansion. The relation of ratios of blood-free to blood-inclusive lungs was different between the two subgroups. Although all regressions were highly correlated, the fits of the blood-free ratio on ratios for blood-inclusive lung and blood together were better than for blood-inclusive lung alone. The mean error of prediction of extravascular lung water for all sheep was significantly less for the regression of blood-free ratio on ratios for blood and blood-inclusive lung together (11 g) than for blood-inclusive lung alone (18 g). This study shows that weights of lung homogenate and blood samples before and after simple oven drying can be used to provide accurate inexpensive estimates of postmortem extravascular lung water.     

The phosphorylation region of lysine-rich histone in dividing cells

N-Bromosuccinimide cleavage of in vivo ³²P-labelled lysine-rich histone isolated from rapidly dividing cells has been studied. N-Bromosuccinimide cleaves F₁-histone into two fragments, a small N-terminal piece and a larger C-terminal portion. The phosphate-induced microheterogeneity and associated radioactivity which has been linked to cell replication, is found in the carboxyterminal fragment. No phosphorous is found associated with the amino-terminal fragment when histone phosphorylation is associated with cell division. The specific tryptic phosphopeptides obtained from in vivo labelled F₁ are clearly different from those obtained from in vitro incubations of free F₁-histone and cytoplasmic protein kinase.     

Further evidence for cortical control over replication of the macronucleus and basal bodies in Stentor

This paper presents further evidence that the cortex controls macronuclear replication and basal body production during the cell cycle of Stentor. At the onset of cell division, basal body production occurs on the ventral side of the cell to form an oral primordium; this structure develops slowly into the oral apparatus destined for the posterior daughter cell. Meanwhile, a series of morphological changes in the macronucleus (coalescence, elongation, nodulation) doubles the number of nodes in preparation for division. When a cell undergoing oral development is grafted to a morphostatic cell of equal size, oral development is usually induced in the morphostatic component and the two members of the graft complex eventually become synchronized with respect to macronuclear morphology. However, primordium induction and nuclear synchronization usually do not occur when the 2 members of the graft complex are separated by cortical discontinuities which heal gradually, even though the graft components demonstrably share a common endoplasm throughout the experiment. These results suggest that the cell surface may control the replication of organelles such as the macronucleus and basal bodies which are normally kept “in step” with the cell cycle in such a way that they are not lost or reproduced too frequently.     

EFFECT OF ECDYSTERONE ON VITELLOGENIN CONCENTRATION IN HAEMOLYMPH OF MALE AND FEMALE SARCOPHAGA BULLATA

Using Polyacrylamide gel electrophoresis, cycloheximide, incorporation of ³H-labelled amino acids and immunological methods, we have demonstrated that injection of ecdy- sterone induces de novo synthesis and release of vitellogenin in both sexes of Sarcophaga bullata. Vitellogenin concentrations were measured by the Mancini-radial immunodiffusion technique. In males a dose as low as 1 ng always makes vitellogenin appear in the haemolymph but very reproducible results are only obtained when doses varying from 10 to 250 ng were injected. In this range, the dose-response curve was linear on a semi- logarithmic scale.

In females, vitellogenin concentration remained low until a few hours after liver feeding and thereafter it rose sharply and reached its maximum about 24 h after the protein meal. 100 μg 6-hydroxydopamine HCl, injected before liver feeding in 4-day-old females, inhibited vitellogenin synthesis and yolk deposition, probably by interfering with the release of a brain hormone. This inhibitory effect on vitellogenin synthesis, but not that on yolk deposition, could be overruled by injection of ecdysterone. Juvenile hormone was ineffective on both. Females, ovariectomized on day 2 or 3, accumulated vitellogenin in their haemolymph, indicating that the continuous presence of the ovaries was not required for vitellogenin synthesis. The possible relation between the gonadotrophs hormone from the brain, vitellogenin synthesis and moulting hormone metabolism is discussed.