An investigation of ethylene inhibition of growth in etiolated Avena sativa coleoptiles /

Growth rates of etiolated Avena sativa coleoptiles in pH 7.0 buffered medium are stimulated in a synergistic manner by IAA and 320 ~l/l carbon dioxide. The suggestion that carbon dioxide stimulated growth involves dark fixation is supported by the ability of 1 mM malate to replace carbon dioxide, with neither factor able to stimulate growth in the presence of the other (Bown, Dymock and Aung, 1974). The regulation of Avena coleoptile growth by ethylene has been investigated in the light of this data and the well documented antagonism between carbon dioxide and ethylene in the regulation of developmental processes. The influence of various permutations of ethylene, IAA, carbon dioxide and malate on the rates of growth, l4c-bicarbonate incorporation, l4C-bicarbonate fixation, and malate decarboxylation have been investigated. In the presence of 320 ~l/l carbon dioxide, 10.8 ~l/l ethylene inhibited growth both in the absence and presence of 20 ~M IAA with inhibition times, of 8-10 and 12-13 minutes respectively. In contrast ethylene inhibition of growth was not significant in the absence of growth stimulation by CO2 or 1 mM malate, and the normal growth increases in response to CO2 and malate were blocked by the simultaneous application of ethylene. The rates of incorporation and dark fixation of l4C-bicerbonate were not measurably. influenced by ethylene, IAA or malate, either prior to or during the changes in growth ,ates induced by these agents. The data does not support the hypothesis that ethylene inhibition of growth results from an inhibition of dark fixation, but suggests that ethylene may inhibit a process which is subsequent to fixation.

Changes in the magnitude and pattern of translocation of photoassimilated ¹CO in soybean plants following an acute exposure to gamma radiation /

Young soybean plants (Glycine ~. L. cultivar Harosoy '63), grown under controlled conditions, were exposed to gamma radiation on a single occasion. One hour following exposure to 3,750 rads, the mature trifoliate leaf of the soybean plant was isolated in a closed system and permitted to photoassimilate approximately 1-5 pCi of 14C02 for 15 minutes. After an additional 45 minute-period, the plant was sacrificed and the magnitude of translocation and distribution pattern of 14C determined. In the non-irradiated plants 18~ of the total 14C recovered was outside the fed leaf blades and of this translocated 14c, 28~ was above the node of the fed leaf, 38~ in the stem below the node, 28~ in the roots and 7~ in the petiole. As well, in the irradiated plants, a smaller per cent (6~) of the total 14 C recovered was exported out of the source leaf blades. Of this translocated 14c , a smaller per cent (20~) was found in the apical region above the node of the source leaf and a higher per cent (45~) was recovered from the stem below the node and in the petiole (11~). The per cent of exported 14 C recovered from the root was unaffected by the radiation. Replacement of the shoot apex with 20 ppm IAA immediately following irradiation, only J partially increased the magnitude of translocation but did completely restore the pattern of distribution to that observed in the non-irradiated plants. From supplementary studies showing a radiationinduced reduction of photosynthetic rates in the source leaf and a reduction of the cumulative stem and leaf lengths in the apical sink region, the observed effects of radiation on the translocation process have been correlated to damage incurred by the source and sink regions. These data suggest that the reduction in the magnitude of translocation is the result of damage to both the source and sink regions rather than the phloem conducting tissue itself, whereas the change in the pattern of translocation is probably the result of a reduced rate of 14C-assimilate movement caused by a radiation-induced decrease of sink metabolism, especially the decrease in the metabolism of the apical sink.

An investigation into differences between indoleacetic acid and fusicoccin in their influence on RNA synthesis, protein synthesis and growth in Avena coleoptile tissue

Growth stimulation of Avena coleoptile tissue by indoleacetic acid (IAA) and fusicoccin (FC) was compared by measuring both their influence on RNA and protein synthesis during IAA or FC stimulated growth. FC stimulated growth more than IAA during the initial four hour exposure, after which the growth rate gradually declined to the control rate. FC, but not IAA, increased the uptake of 3H-Ieucine into tissue and the specific radioactivity of extracted protein. Cycloheximide inhibited the incorporation of 3H-Ieucine into protein by approximately 60% to 70% in all cases. In the presence of cycloheximide 3H-radioactivity accumulated in FC-treated tissue, whereas IAA did not seem to influence 3H-accumulation. These results suggest that FC stimulated leucine uptake into the tissue and that increased specific activity of coleoptile protein is due to increased leucine uptake, not an increased rate of protein synthesis. There was no measurable influence of IAA and/or FC on RNA and protein synthesis during the initial hours of a growth stimulation. Inhibitors of RNA and protein synthesis, actinomycin D and cycloheximide, respectively, severely inhibited IAA enhanced growth but only partially inhibited FC stimulated growth. The data are consistent with suggestions that a rapidly turning over protein participates in IAA stimulated growth, and that a continual synthesis of RNA and proteins is an absolute requirement for a long term growth response to IAA. On the contrary, FC-stimulated growth exhibited less dependency on the transcription and translation processes. The data are consistent with proposals suggesting different sites of action for FC and IAA stimulated growth. l?hen compared to CO2-free air, CO2 at 300 ppm had no significant influence on coleoptile growth and protein synthesis in the presence or absence of lAA or FC. Also, I mM malate, pH 6.0 did not influence growth of coleoptiles in the presence or absence of lAA. This result was obtained despite reports indicating that 300 ppm CO2 or I mM malate stimulates growth and protein synthesis. This lack of difference between CO2-treated and untreated tissue could indicate either that the interstitial space CO2 concentration is not actually different in the two treatments due to significant endogenous respiratory CO2 or else the data would suggest a very loose coupling between dark CO2 fixation and growth. IAA stimulated the in vivo fixation of 14c-bicarbonate (NaHI4c03) by about 25% and the addition of cycloheximide caused an inhibition of bicarbonate fixation within 30 min. Cycloheximide has also been reported to inhibit IAA-stimulated H+ excretion. These data are consistent with the acid growth theory and suggest that lAA stimulated growth involves dark CO2 fixation. The roles of dark CO2 fixation in lAA-stimulated growth are discussed.

The influence of carbon dioxide on growth and metabolism of etiolated Avena sativa 1 coleoptiles

The influence of carbon dioxide on growth and protein synthesis of etiolated Avena coleoptiles was investigated. Evidence is presented that 0.03% carbon dioxide stimulated both these processes; and that carbon dioxide stimulated growth depends on carbon dioxide stimulated protein synthesis, In addition the evidence indicates that carbon dioxide stimulated growth is mediated by metabolism, and that carbon dioxide stimulates growth through a dark fixation process. Growth studies also demonstrated that IAA and carbon dioxide stimulated growth in a synergistic manner.