Induction of Solute Release from Nicotiana tabacum Tissue 10

For determination of the effects of polymyxin B, polymyxin E,or ethylenediamine tetra-acetic acid (EDTA) on plant cell membranes,the rates at which three solutes, K⁺, P₁, and sugar, leakedfrom treated tissue culture cell suspensions of Nicotiana tabacumwere measured. The kinetics of leakage from cells treated witheither of the polymyxins was biphasic, whereas kinetics forcells treated with EDTA was monophasic. Only K⁺ leaked frompolymyxin-treated cells during the first phase, and all threesolutes leaked during the second phase. The slower first phaseis interpreted as leakage of K⁺ from the Donnan free space andcytoplasm, and the faster second phase as the leakage of solutesfrom the vacuole. The monophasic kinetics of EDTA treatmentindicated that solutes were leaking simultaneously from cytoplasmand vacuole. Of the divalent cations tested, only Ca⁺⁺ and Mn⁺⁺counteracted the effects of polymyxin and EDTA. Ca⁺⁺ even restoredP¹ and sugar uptake. Addition of Mg⁺⁺ or Sr⁺⁺ to polymyxin-treatedcells did not stop solute leakage but actually enhanced theleakage rates. A model is presented that suggests that polymyxinor EDTA induces solute leakage by forming pores in plant cellmembranes. The effects of divalent cations on membranes oncethe pores are formed are also discussed. Key words: Polymyxin, EDTA, Nicotiana tabacum, Solute leakage     

ADENOSINE TRIPHOSPHATASE ACTIVITY IN GERMINATING LETTUCE SEEDS

Mitochondria isolated from lettuce seeds and seedlings are capableof hydrolyzing ATP. On basis of pH optima, there are at leasttwo and probably three enzymes possessing ATPase activity. Oneof them is inhibited by EDTA and all of them are inhibited byNaF. The ATP ase activity of the mitochondria is very slight in imbibedseeds but increases considerably with seedling growth. Othercomponents of the cell also contain ATP hydrolyzing enzymes;however, most of these phosphatases in the seedling seem tobe soluble enzymes. (Received December 12, 1961; )     

COMPARISON OF THE OSMOTIC VOLUME CHANGES IN CHLOROPLASTS ISOLATED FROM DIFFERENT PLANTS

Chloroplasts were isolated from leaves of four plant species(spinach, New. Zealand spinach, Swiss Chard and tomato) andtheir osmotic behaviour was compared. Maximal contraction ofchloroplasts from all plant sources occurred at 0.3–0.4M sucrose. The volume of the particles at this sucrose concentrationwas also similar in all cases; 300 mm³ per 10¹⁰ chloroplasts.At higher and lower sucrose concentrations, the chloroplastsswell. The degree of swelling, especially in higher sucroseconcentrations, differed in chloroplasts from different sources.Tomato chloroplasts are most sensitive, and the spinach chloroplastsare the most resistant to all types of treatment. Tomato chloroplastsrequire rather high phosphate concentrations for good preservation. Changes in optical density at 520 mµ can be used to geta rough idea about the condition of the particles. (Received February 7, 1966; )     

PROPERTIES AND CHANGES OF GLUCOSE-6-PHOSPHATE DEHYDROGENASE IN GERMINATING LETTUCE SEEDS

Glucose-6-phosphate dehydrogenase is present in lettuce seedsand seedlings in a particulate, 20,000 g fraction and in a solublefraction of the cells. In both fractions the enzyme activityoccurs in the presence of either NAD or NADP. It seems probablethat an enzyme with two active sites is involved. Total activity does not change during 48 hours germination inwater. It is depressed by coumarin or thiourea treatment. Specificactivity rises in the soluble fraction when germination occurs,but not in the mitochondrial one. Coumarin treatment largelydepresses the increase in specific activity. In vitro 100 ppmcoumarin totally inhibit enzyme activity. 1,250 ppm thioureado not affect enzyme activity. The results are analysed and it is concluded that glucose-6-phosphatedehydrogenase is important, if at all, only in the very earlystages of germination. (Received August 13, 1965; )     

Interaction Between Mineral Nutrients, Cytokinin and Gibberellic Acid During Growth of Sorghum at High NaCI Salinity

Sorghum bicolor (L.) Moench, cv. 610, adapted to high salinitywas able to grow at 300 mol m^–3 NaCl only when half-strengthHoagland's solution was enriched with mineral nutrients. Theoptimal growth rate was observed in full strength Hoagland'ssolution; at higher or lower concentrations growth rates werelower. In contrast, growth rate of plants exposed to 150 molm^–3 NaCl was not affected by similar modification of theHoagland solution concentration. At high salinity, additionof cytokinin (CK) or gibberellic acid (GA), or a mixture ofboth, can induce the same effect on growth as the increasedmineral nutrient concentration. Phytohormones and increasedmineral concentration have similar effects, possibly becausean imbalance in phytohormones, rather than a mineral deficiency,limits growth at 300 mol m^–3 NaCl in the presence of half-strengthHoagland solution. The change in mineral concentration in thenutrient medium, in addition to its nutritional effect, alsoapparently acts as a signal involved in hormonal balance whichallows growth at high salinity. Exposure of Sorghum to 300 molm^–3 NaCl causes a decrease in the range of nutrient concentrationswhich can sustain growth. Adjustment of the nutrient concentrationmay induce the synthesis of endogenous CK and GA concentrationsrequired for growth. In contrast, addition of CK or GA at similarconcentrations during the adaptation (pretreatment) period inhibitsgrowth and prevents the adaptation process. The response tothe exogenous phytohormone treatments depends on the time elapsedfrom the beginning of salinization. Key words: Adaptation to salinity, cytokinin, gibberellic acid, mineral nutrition, growth, Sorghum, NaCl     

Exogenous ABA as a Modulator of the Response of Sorghum to High Salinity

Treatment of Sorghum bicolor (L.) Moench, cv. 610, with abscisicacid (ABA) during the first week of sahnization with 150 molm^–3 NaCl induced enhancement of growth and acceleratedadaptation to high salinity (300 mol m^–3 NaCl) Adaptationis defined as the development of the ability of the plant tosurvive, grow, and set seeds upon exposure to a NaCl concentrationwhich is lethal for the unadapted plant In the absence of ABAthe saline pretreatment requires 20 d for the development ofadaptation (Amzallag et al., 1990), with ABA treatment the sameresult is achieved within approximately one week The exposureof the plants to non-lethal salinity (150 mol m^–3 NaCl)apparently triggers a transient sensitivity to ABA lasting forabout 8 to 10 d following the beginning of sahnization Thisperiod coincides with an increase in leaf PEP carboxylase activitywhich seems to occur faster if the plants are treated with ABA.Exogenous ABA-induced enhancement of growth and control of shootNa⁺ concentration, occur at a lower ABA concentration (10 mmolm^–3) than the induction of adaptation to salinity whichoc curs at 40 mmol m^–3 or above. The lowered shoot Na⁺concentration which is induced by a low ABA concentration isnot sufficient to induce survival of the plants in high salinity(300 mol m^–3 NaCl). Key words: Growth, adaptation to salinity, ABA     

Some Quantitative Changes in the Fat Metabolism of Germinating Lettuce Seeds and their Inhibition by Coumarin

The fat reserve of lettuce seeds of two varieties, ‘GrandRapids’ and ‘Progress’, was examined. It wasfound to be an unsaturated lipid, closely resembling sunflower-seedoil. The changes in the fat content with time and under treatmentwith thiourea and with coumarin were studied, as were the changesin the acidity of the fat. No free volatile saturated acidswere detected. On hydrolysis the seeds yield one such acid whoseidentity was not finally established and whose origin is uncertain,it not being certain that it is connected directly with fatmetabolism.     

Ion Absorption and Allocation of Carbon Resources in Excised Pea Roots Grown in Liquid Medium in Absence or Presence of NaCl

Freshly excised pea roots (Pisum sativum cv. Alaska) when transferredto growth medium (130 mOsm) or growth medium containing salt( 370 mOsm) suffer an initial osmotic shock and lose water.Contol roots tended to accumulate potassium, particularly inthe apical zone, while those exposed to NaCl accumulated mainlysodium, potassium accumulation being depressed. Exposure tosalinity for 6 d caused increases in root protein, cellulose,uronic acid and lignin content per cell. In roots supplied with¹⁴C-glucose for 24 h immediately after excision there was littledifference in uptake of glucose and in its use in respirationbetween control and salt treated roots. However, there werenoticeable differences in incorporation of labelled carbon intoseveral cell fractions, and particularly into the cellulosefraction in the upper parts of the root. When roots were grownfor six days in culture before being supplied with [¹⁴C]glucose,uptake per root was greater in the 120 mM NaCl treatment, andthe fraction diverted to respiration was decreased by salinity.On a per cell basis incorporation into soluble starch, uronicacid and cellulose fractions was increased in the salt treatedroots. The data obtained are in accord with the previous findingsand are suggestive of increased synthesis of cell wall materials.No conclusion could be drawn as to whether the changes describedare of adaptive value. Pisum sativum L. cv. Alaska, root culture, salinity, osmoregulation, cell wall     

Lactic acid content and formation in pea roots exposed to salinity

Lactic acid content and production, as well as lactic dehydrogenaseactivity were studied in pea root tips grown in media salinatedwith NaCl or Na₂SO₄. Salinity of both types depressed the lacticacid content of the tissue. Lactic dehydrogenase activity linkedto NADH was depressed by high concentrations of NaCl in thegrowth medium, but was stimulated by increasing concentrationsof Na₂SO₄. There was a trace of NADPH linked activity of theenzyme; this activity was not affected by chloride salinitybut was depressed by sulphate salinity. It is not yet clearhow these events are connected with the overall effect of salinityon the carbohydrate metabolism of pea root tips. (Received June 27, 1970; )