Effect of {alpha}-Tomatine on the Response of Wheat Coleoptile Segments to Exogenous Indole-3-acetic Acid

A concentration of 10^–5 M tomatine had no effect on leakagefrom, or elongation of, wheat coleoptile segments, but consistentlyreduced IAA-enhanced extension growth by c. 50 per cent. Therewas no evidence of chemical interaction between the alkaloidand the auxin in solution, and IAA action was not affected bypre-treatment for up to 3 h with 10^–5 M tomatine. Studieswith [2-¹⁴C]IAA revealed that 10^–5 M tomatine did notinhibit uptake of auxin into segments. The effect of pre-treatingsegments for up to 3 h with IAA could be virtually nullifiedby 10^–5 M tomatine, as could also IAA-induced changesin properties of coleoptile cell walls. Results are discussedin relation to the ability of tomatine to disrupt membrane functionand to current hypotheses implicating membranes in the primaryaction of auxin.     

Response of Tissues2

Efflux of K⁺ from tomato fruit pericarp disks was not affectedby 100 µM tomatine, and only slightly by 1 mM. No differentialresponses to the alkaloid were observed in disks obtained fromfruits at different stages of development, and there was noevidence of any correlation between the level of endogenoustomatine and the effect of exogenous tomatine. Growth of tomatoseedlings in liquid nutrient medium was unaffected by 1 mM tomatinewhereas the growth oflettuce seedlings was stunted in the presenceof 100 µM alkaloid. The polyene antibiotic nystatin gavesimilar resultsto tomatine, having little effect on tomato tissuesbut causing marked inhibition of lettuce seedling development.     

The NADP$-specific isocitrate dehydrogenase of Rhodospirillum rubrum

The NADP^$-specific isocitrate dehydrogenase was partially purifiedfrom photosynthetically-grown Rhodospirillum rubrum. The pHoptimum is between 7.5 and 9.0 in phosphate buffer. The apparentKm is 3.1x10^–5 M for isocitrate, 5.1x10^–5 M forNADP^$, 1.7x10^–5 M for manganese, 1.5x10^–4 M formagnesium, and 3.5x10^–3 M for inorganic orthophosphate.Arsenate exerts a slight inhibition. The Q₁₀ between 17.5°Cand 40°C is 1.62, and the energy of activation at 25°Cis 9.74 Kcal/mole. Glyoxylate and oxalacetate cause concertedinhibition of the enzyme activity. Various nucleotides inhibitthe activity. The kinetics of inhibition by ATP was found tobe mixed type with respect to NADP^$ and isocitrate, the K_i valuesbeing 1.17x10^–3 M and 1.10x10^–3 M respectively.The inhibition between ATP and orthophosphate is competitivewith a K_i of 10^–4M. Thiol binding reagents are inhibitory;this inhibition is reversed by cysteine or reduced glutathione. (Received October 1, 1971; )     

Environmental and hormonal control of turion formation in Myriophyllum verticillatum

The turions of Myriophyllum verticillatum, an aquatic vascularplant, develop in the fall and function in propagation and dispersalas well as in over-wintering. Experiments with controlled evnironmentsindicate that both temperature and photoperiod regulate turionformation. Turions can be induced at 15°C or lower, butnot at 20°C. At 15°C, turions form in both 8- and 12-hrdays, but not in 16-hr days. Plants collected in early springdo not form turions readily in response to short days unlesspreviously exposed to long days; thus, turion formation is along-day-short-day response. This combination of photoperiodand temperature requirements probably prevents turion developmentin early spring when the temperature and photoperiod are similarto those in the fall. Treatment of plants with ABA (10^–5M) enhances turion development under marginally inductive conditions(12-hr days at 15°C) but cannot induce it under long days.On the other hand, the cytokinin benzyladenine (10^–5 M)blocks turion formation. GA3 (10^–5 M) and AMO-1618 (10^–5M) exert only small qualitative effects on turion development,while IAA (10^–5 M) retards it. During turion development,the level of ABAlike activity and of one or two unidentifiedinhibitors increases. Cytokinin activity decreases at the startof turion formation, increases during development, then decreasesat abscission. Thus two lines of evidence suggest that a decreasein cytokinin activity and an increase in acidic inhibitor activityplay important roles in turion induction. ¹Present address: Biological Station, University of Michigan,Ann Arbor, Michigan 48109, U. S. A. (Received December 1, 1975; )     

Effects of xylopine on physiological activities of Scenedesmus obliquus

Xylopine, an aporphine alkaloid contained in the bark of Xylopiadiscreta strongly inhibits growth of Scenedesmus obliquus; 50%inhibition was caused by 2x10^–5 M of xylopine. Photosynthesis was also strongly inhibited by xylopine at concentrationsabove 10^–4 M. Effects on photosynthesis, however, differeddepending on the ratio of alkaloid concentration to cell concentrationin the reaction mixture, so that under certain experimentalconditions with concentrations of xylopine below 10^–4M, there was some acceleration of photosynthesis. Xylopine strongly inhibited the HILL reaction with benzoquinoneas the HILL oxidant. Respiration was not affected at concentrationscompletely inhibiting photosynthesisd growth of the alga. A probable role of the aporphine alkaloid in controlling lichengrowth in nature is discussed. (Received March 25, 1970; )     

Enzymes involved in the last steps of the biosynthesis of mannitol in brown algae

Mannitol-1-phosphate dehydrogenase (EC 1.1.1.17 [EC] ) and mannitol-1-phosphatase(EC number yet unassigned) were detected in the brown algae,Spatoglossum pacificum and Dictyota dichotoma. The enzymes wereextracted from algal fronds and their properties were investigatedusing partially purified preparations. Mannitol-1-phosphatase shows maximum activity at pH 7. The enzymehad a narrow substrate specificity. The Km value for mannitol-1-phosphateis 8.3x10^–4 M (30°C, pH 7.0). The enzyme is activatedby Mg⁺⁺ and Mn⁺⁺and is strongly inhibited by PCMB, Hg⁺⁺and NaF. Mannitol-1-phosphate dehydrogenase showed maximum activitiesat pH values 6.5 and 10.2 in reductive and oxidative reactions,respectively. The dehydrogenase also showed narrow substratespecificity; mannitol-1-phosphate and NAD or fructose-6-phosphateand NADH₂ are utilized, respectively, in oxidative and reductivereactions by the enzyme. Km values for these substrates andthe coenzymes are 2.5x10^–4 M and 7.1x10^–5 M forthe first pair and 2.8x10^–4 M and 1.3x10^–5 M forthe latter pair. This enzyme was strongly inhibited by PCMBand Hg⁺⁺, but was only slightly affected by adenosine phosphates. Possible roles of these enzymes in the biosynthesis of mannitolin brown algae are discussed. ¹ Contributions from the Shimoda Marine Biological Station ofTokyo Kyoiku University, No. 233. This work was supported inpart by a Grant-in-Aid for Co-operative Research from the Ministryof Education, Japan and in part by a grant to one of us (T.Ikawa) from the Matsunaga Science Foundation. ² Present address: Chemical and Physical Laboratory, HoechstJapan Research Laboratory, Minamidai, Kawagoe, Japan. (Received February 22, 1972; )     

Effects of Anoxia on Wheat Seedlings: I. INTERACTION BETWEEN ANOXIA AND OTHER ENVIRONMENTAL FACTORS

Anoxia was imposed on 4–6-d-old, intact wheat seedlings,after the roots had first been exposed for 1 d to O₂ concentrationsbetween 0·016 and 0·06 mol m^–3. Apices ofthe main axis of the seminal roots were considered to have toleratedanoxia if elongation occurred after return from anoxia to air,hereafter called ‘retention of elongation potential’.During anoxia, elongation potential was retained longer in rootsof intact seedlings than in 0–5 mm excised root tips suppliedwith 50 mol m^–3 glucose. In intact seedlings, elongation potential was retained longerat 15°C than at 25°C, and at pH 50 and 60 than at pH40. These differences between treatments were maintained inthe presence of exogenous glucose, and glucose supply prolongedthe retention of elongation potential in all anoxic treatments. Elongation potential was retained much longer at very low 02concentrations (0006 to 00l mol m^–3) than under anoxia;this was established at pH 40. Anoxia inhibited the transport of sugars from the shoots and/orendosperm to the root by 79-97%, as assessed from experimentswith roots of intact plants exposed to anoxia at pH 60 and 15°C. Overall, the results demonstrate: (i) that the occurrence ofadverse effects of anoxia during waterlogging in the field mayinteract with other environmental factors and (ii) that thereare pronounced difficulties integrating data on tolerance toanoxia obtained in different laboratories. Key words: Anoxia, wheat seedlings, pH, temperature     

Effects of Temperature and dl-Strigol on Seed Conditioning and Germination of Witchweed (Striga asiatica)

Seed conditioning and germination in witchweed (Striga asiatica(L.) Kuntze) were temperature-dependent. With higher conditioningtemperatures, shorter conditioning time was required for germinationwith terminal dl-strigol (strigol) treatment at 30 °C. Maximumgermination (80–100%) was obtained by conditioning inwater at 20, 25, 30 and 35 °C for 14, 7, 5 and 3 d, respectively,and terminally treating with 10^–6 M strigol at 30 °C.Seeds conditioned in 10^–8 M strigol instead of water germinatedmuch less with the same terminal strigol treatment. Generally,conditioning was slower when seeds were conditioned in strigolrather than water. The reduction in germination rate by pretreatmentin strigol or pretreatment at low temperatures could be overcomeby increasing the terminal strigol concentration in the germinationtest. Conditioned seeds did not germinate at 10 and 15 °Cwith a terminal 10^–6 M strigol treatment but yielded closeto maximum germination at 25, 30 and 35 °C with the sameterminal strigol treatment. To obtain maximum germination, boththe minimum conditioning temperature and the minimum germinationtemperature for conditioned seeds were 20 °C. Factors suchas conditioning time, and strigol concentration and temperatureduring conditioning and/or germination determine whether seedsremain in the conditioning phase or shift to a germination phase. dl-Strigol, germination stimulation, parasitic plants, seed conditioning, seed germination, Striga asiatica, temperature, weed control     

Light-induced changes in membrane potential in Spirogyra

Spirogyra cells exhibited changes in membrane potential whenthey were exposed to light. Cells made chloroplast-free didnot show any light-induced potential change (LPC) upon illuminationwith white light and also monochromatic red (680 nm) and farred (720 nm) light. LPC was observed when the cell containedonly a small fragment of chloroplast, whether the cell had anucleus or not. The magnitude of LPC depended on the amountof chloroplast in the cell. DCMU at 10^–5 M, CCCP at 10^–5 M and DNP at 10^–4M at pH 5.5 suppressed LPC, while CCCP at 1–5 ? 10^–6M, NH₄Cl at 5 ? 10^–2 M and DNP at 10^–4 M at pH 7.0stimulated LPC. PMS at 10^–4 M stimulated LPC and couldinduce LPC which was completely inhibited by DCMU. These factssuggest that LPC is related to noncyclic and cyclic electronflows. The influences of light and dark conditions and various metabolicinhibitors (DCMU, DNP, CCCP, NH₄Cl) on ATP level have been investigated.No significant difference in the ATP level was observed betweencells in the light and dark. DNP at 10^–4 M (pH 5.5) andCCCP at 5 ? 10^–6 M decreased the ATP level significantly,while DCMU and NH₄Cl only slightly. Good correlation was notfound between the total ATP level and LPC in Spirogyra. LPC occurred even when the external medium contained only asingle salt such as KCl, NaCl or CaSO₄. LPC was also recorded in chloroplasts in situ and in vitro.The mode of LPC of chloroplasts was quite different from thatof the cell. On illumination, the chloroplast potential changedvery rapidly and transiently in the positive direction thenrecovered spontaneously to almost the original potential level. Possible causes of LPC are discussed in relation to the electrogenicion pump. ¹ Present address: Department of Botany, Faculty of Science,University of Tokyo, Hongo, Bunkyo, Tokyo 113, Japan. (Received November 9, 1977; )     

Effect of temperature and food concentration on post-embryonic development, egg production and adult body size of calanoid copepod Eurytemora affinis

Post-embryonic development time, egg production rate and adultbody size of calanoid copepod Eurytemora affinis from Lake Ohnuma,Japan, were determined under six temperature-food conditions(10³,5 x 10³, 10⁴ and 5 x 10⁴ cells ml^–1 at 15°C,5 x 10⁴ cells ml^–1 at 10and20°C) in the laboratory.The measured parameters varied with both temperature and foodconcentration. Development time from hatching to adult femalewas 9.2, 11.4 and 22.8 days at 20, 15 and 10°C respectively,at the highest food concentration. The males developed to adultat one to two days earlier than the females. An effect of foodshortage on development time occurred at the lowest food concentration.This development time was 24.8 days even at 15°C, beingtwice as long as that at the highest food concentration. Prosomelength of these food-limited females was approximately 75% ofwell-fed ones, which reduced by only 10% with increasing temperaturefrom 10 to 20°C. Clutch size (C, eggs clutch^–1) ofwell-fed individuals depended on prosome length of the adultfemale (L, mm), and was expressed as an equation: C = 65.2 L³⁸³. Clutch size of individuals reared at less than 10⁴ cellsml^–1, however, mostly laid below the estimated curve,especially at the lowest food concentration being only 10% ofthat at the highest food concentration. These results suggestthat food availability is a more important factor affectingpopulation growth of E.affinis in Lake Ohnuma than variationof temperature.     

Control of Plasma Membrane Cl- Fluxes in Chara corallina by External Cl- and Light

The efflux of Cl^– at the plasma membrane of Chara wasstudied in relation to two treatments known to affect the flux:that of removal of external Cl^– and of light. It is shownthat although removal of external Cl^– results in a rapidreduction in Cl^– efflux (consistent with a direct effectof external Cl^– on the transport system) the magnitudeof this reduction in the dark is greater than the measured darkinflux. Therefore, in the dark at least, it is proposed that1:1 exchange diffusion cannot account for the trans-stimulationof efflux by external Cl^–. Light induces an inhibition of efflux and a concomitant stimulationof Cl^– influx at 20 °C, but at 10 °C the responsesto light of the two fluxes can be separated temporally. It istherefore suggested that the fluxes are not reciprocally dependenton the same factor which mediates in the light response. Furtherconsiderations show that it is unlikely that the cytoplasmicCl^– concentration mediates in the light response of eitherflux, but that changes in cytoplasmic pH may do so.     

Studies on NADP-isocitrate dehydrogenase from maturing castor bean seeds

NADP-specific isocitrate dehydrogenase from the soluble fractionof maturing castor bean endosperm was partially purified (approximately180-fold) and some of its enzymatic properties were studied.Mg⁺⁺, Mn⁺⁺, Cd⁺⁺, Ba⁺⁺, Co⁺⁺, Zn⁺⁺, and Sr⁺⁺ were activatorsof the enzyme reaction at a concentration of 6.7x10 M. The optimumpH of this enzyme was about 8.5. The enzyme was stable in thenarrow range from pH 7.0 to pH 8.0. Km values for isocitrateand NADP at pH 8.5 were 3.5x10^–6 M and 3.6x10^–6M, respectively. Enzyme stability was not affected by NaCl concentrationand enzyme reaction was inhibited at 5x10^–6 M PCMB (80%inhibition). It is suggested that the condensation product ofglyoxylate and oxalacetate also inhibits the reaction. NADP-IDHin the crude extract from maturing castor bean endosperm washeat-stable but the dialyzed enzyme preparation and the partiallypurified enzyme were labile against heat treatment at 57°C.When Mg⁺⁺ was added to the partially purified enzyme in thepresence of isocitrate or NADP, the enzyme was stabilized againstheat treatment. Mn⁺⁺, Ca⁺⁺, Co⁺⁺, Sr⁺⁺ or Ba⁺⁺ could be substitutedfor Mg⁺⁺. Addition of only one of the factors, Mg⁺⁺, isocitrateor NADP, had no effect on the heat stability. Moreover, a combinationof isocitrate and NADP did not establish stabilization. A divalentcation plays a central role, while adenine nucleotide, especiallyATP, may have an important part in stabilization. (Received August 14, 1972; )     

Acclimation of NO-3 fluxes to low root temperature by Brassica napus in relation to NO-3 supply

Acclimation of NO^–₃ transport fluxes (influx, efflux)in roots of oilseed rape (Brassica napus L. cv. Bien venu) andtheir sensitivity to growth at low root temperature was studiedin relation to external NO^–₃ supply, defined by constantconcentrations ranging from sub- to supra-optimal with respectto plant growth rate. Plants were grown from seed in flowingnutrient solutions containing 250 mmol m^–3 NO^–₃at 17°C for 20d, and solution temperature in half the cultureunits was then lowered decrementally over 3 d to 7°C. Threedays later plants were supplied with NO^–₃ at 1, 10, 100or 1000 mmol m^–3 maintained for 18 d. Dry matter productionwas decreased more by low root zone temperature than low [NO^–₃]_e. Root specific growth rates were inversely related to [NO^–₃]_eand shoot:root ratios increased with time at [NO^–₃]_e between10–1000 mmol m^–3. Net uptake of NO^–₃ at 17°Cwas twice that at 7°C, and at both temperatures it doubledwith increasing [NO^–₃]_e between 1–10 mmol m^–3with further small increases at higher [NO^–₃]_e. Mean unitabsorption rates of NO^–₃ between 0–6 d and 6–14d were linearly related (r² of 0.79–0.99) to log₁₀[NO].Steady-state Q₁₀ (7–17°C) for uptake between 0–6d were 0.91, 1.62, 1.27, and 1.10, respectively, at [NO^–₃]_eof 1, 10, 100, and 1000 mmol m^–3, compared with correspondingvalues of 0.98, 1.38, 1.68, and 1.89 between 6–14 d. Thedata indicated that net uptake rates at 7 and 17°C divergedover time at high [NO^–₃]_e. Short-term uptake rates from1 mol m^–3 NO^–₃ measured at 17°C were higherin plants grown with roots at 7°C than at 17°C; for7°C plants there was a strong inverse linear relationship(r²=0.94) between uptake rate and treatment log₁₀ [NO^–₃]_ewhilst rates in 17°C plants were independent of prior [NO^–₃]_e. Rates of NO^–₃ influx and efflux under different steady-stateconditions of NO^–₃ supply and root temperature were calculatedfrom dilution of ¹⁵N added to culture solutions. Efflux wassubstantial relative to net uptake in all treatments, and wasinversely related to [NO^–₃]_e at 17°C but not at 7°C.Ratios of influx: efflux ranged from 1.6–2.9 at 17°Cand 1.3–1.8 at 7°C, indicating the proportionatelygreater impact of efflux at low root temperature. Ratios ofefflux: net uptake were 0.53–1.56 at 17°C and 1.21–3.58at 7°C. The apparent sensitivities of influx and effluxto steady-state root temperature varied with [NO^–₃]_e.Both fluxes were higher at 17°C than 7°C in the presenceof 100–1000 mmol m^–3 NO^–₃ but the trend wasreversed at 1–10 mmol m^–3 NO. Concentrations oftotal N measured in xylem exudate were at least 2-fold higherat 7°C compared with 17°C, attributable mainly to higherconcentrations of NO^–₃ glutamine and proline. The resultsare discussed in terms of acclimatory and other responses shownby the NO^–₃ transport system under conditions of limitingNO^–₃ supply and low root temperature. Key words: Brassica napus, nitrate supply, efflux, influx, root temperature, xylem exudate     

Involvement of cellulose synthesis in actions of gibberellin and kinetin on cell expansion. Gibberellin-coumarin and kinetin-coumarin interactions on stem elongation

In azuki bean (Azukia angularis = Vignia angularis) epicotylsections, 5 ? 10^–4 M coumarin inhibited the incorporationof radioactivity from [U^–14C]glucose into the cellulosefraction by 35% in the absence of indole-3-acetic acid (IAA)and by 40% in the presence of 1 ? 10^–4 M IAA. There wasno inhibitory effect on the incorporation of radioactivity intothe other fractions. Coumarin at 5 ? 10^–4 M reversed thepromoting effect of 1 ? 10^–5 M gibberellin A₃ (GA) andthe inhibitory effect of 1 ? 10^–5 M kinetin on IAA-inducedelongation of sections with no significant effects on IAA-inducedelongation. Neither GA nor kinetin had any appreciable effectson cellulose synthesis. No inhibition of cellulose syntheiswas observed with 1 ? 10^–3 M colchichine, which has beenreported to have effects similar to those of coumarin on GA-or kinetin-affected stem elongation. Coumarin at 5 ? 10^–4M was ineffectual in breaking up wall microtubules, while adisrupting effect on wall microtubules was clearly demonstratedwith 3 ? 10^–4M colchicine. From these results, the possible involvement of cellulose synthesisin cell expansion controlled by GA or kinetin was suggested. (Received August 3, 1973; )     

ABA and Low Temperature Induce Freezing Tolerance via Distinct Regulatory Pathways in Wheat

The role of ABA in the induction of freezing tolerance was investigatedin two wheat (T. aestivum L.) cultivars, Glenlea (spring var)and Fredrick (winter var). Exogenous application of ABA (5x10^–5M for 5 days at 24°C) increased the freezing tolerance ofintact plants by only 3°C (LT₅₀) in both cultivars. Maximalfreezing tolerance (LT₅₀ of –9°C for Glenlea and –17°Cfor Fredrick) could only be obtained with a low temperaturetreatment (6/2°C; day/night) for 40 days. These resultsshow that exogenously applied ABA cannot substitute for lowtemperature requirementto induce freezing tolerance in intactwheat plants. Furthermore, there was no increase in the endogenousABA level of wheat plants during low temperature acclimation,suggesting the absence of an essential role for ABA in the developmentof freezing tolerance in intact plants. On the other hand, ABAapplication (5x10^–5 M for 5 days at 24°C) to embryogenicwheat calli resulted in an increase of freezing tolerance similarto that achieved by low temperature. However, as in intact plants,there was no increase in the endogenous ABA level during lowtemperature acclimation of calli. These results indicate thatthe induction of freezing tolerance by low temperature is notassociated with an increase in ABA content. Using an antibodyspecific to a protein family associated with the developmentof freezing tolerance, we demonstrated that the induction offreezing tolerance by ABA in embryogenic wheat calli was correlatedwith the accumulation of a new 32 kDa protein. This proteinis specifically induced by ABA but shares a common antigenicitywith those induced by low temperature. These results suggestthat ABA induces freezing tolerance in wheat calli via a regulatorymechanism different from that of low temperature. (Received June 15, 1993; Accepted September 16, 1993)     

The Osmotic Pressure of Concentrated Solutions of Polyethylene Glycol 6000, and its Variation with Temperature

The vapour pressures of aqueous solutions of polyethylene glycol6000 have been measured (by equilibration with sucrose solutions)up to the saturation point at 25 °C (1.45 g g^–1 water).The reduced-osmotic-pressure (/c), when plotted versus concentration(c), rapidly and linearly increased up to a concentration ofabout 0.8 g g^–1 (crossing the similar plot for sucrose).Above this concentration, the reduced-osmotic-pressure rosemore slowly, but still more rapidly than sucrose. The maximumosmotic pressure achieved at saturation was nearly 18 MPa. Usingthe virial equation: /c= RT/M + RTA₂c, the calculated secondvirial coefficient (A₂) for the linear part is 4.5 x 10^–3mol g^–1, a value slightly greater than most literaturevalues at 25 °C. Data are cited showing that A₂ varies linearlyfrom 5–6 x 10^x3 at 0 °C, to zero at 80–90 °C     

Carbon Translocation in the Tomato: Pathways of Carbon Metabolism in the Fruit-

The rate of carbon import by tomato fruits has been relatedto their carbon metabolism by examining the effects of fruittemperature on the metabolism of imported assimilates. ¹⁴C–sucrose,–glucose, –fructose, –malic acid and –citricacid were injected individually into young growing tomato fruitswhich were subsequently maintained at 25 or 5 °C for 48h. Fruit temperature greatly affected the proportions of ¹⁴Clost from the fruits by export and respiration. Only 40 percent of the injected ¹⁴C from ¹⁴C–sugars and 20 per centfrom ¹⁴C–acids was recovered from fruits at 25 °C.Less than 10 per cent of the injected ¹⁴C was exported, thebalance being respired. In contrast, more than 50 per cent ofthe injected ¹⁴C was recovered from cooled fruits, in whichthe import rate of carbon was presumably reduced, and 20–36per cent of injected ¹⁴C was exported. Cooling enhanced thesynthesis of ¹⁴C–sucrose from injected ¹⁴C–hexosesand inhibited the incorporation of ¹⁴C into starch and insolubleresidue. When ¹⁴C–sugars were injected, radioactivityexported from the cooled fruits was detected as sucrose in thephloem of the peduncles; radioactivity was also detected instems and roots when fruits were cooled. In almost fully–grownfruits injected ¹⁴C–compounds were metabolized less readilythan in smaller fruits. Conversion of ¹⁴C–hexoses to ¹⁴C–sucrosewas again enhanced by cooling (5 °C, but was less in fruitsmaintained at 35 °C than in controls. Lycopersicon esculentum, tomato, fruit, translocation, carbon metabolism     

Calcium Involvement In the IAA-induced Leaflet Opening of Cassia fasciculata

Opening of Cassia fasciculata leaflets was induced in darknessafter application of indole-3-acetic acid (IAA). This movementwas obtained with concentrations from 10^–6 M to 10^–4M, after a corresponding time-lag ranging from 120 to 30 min.IAA (5x10^–5 M) allowed leaflet opening at all the pH valuestested (from 3·5 to 7·5), the largest aperturebeing obtained at pH 60 in MES 2·5 mM. Our data suggesta functional involvement of calcium in the regulation of theturgor variations occurring in the pulvinar motor cells duringIAA-induced leaflet opening which occurs in darkness: indeed,this movement was inhibited by the Ca²⁺ chelator EGTA (thisinhibition was reversed by CaCl²) or by antagonists (LaCl³,TMB-8); on the contrary, the IAA-opening was enhanced by ionophoreA 23187. Calcium mobilization through specific channels was tested usingantagonists such as verapamil and nifedipine: at physiologicaldoses, these compounds did not significantly affect leafletresponse. The possibility that calcium could originate frominternal stores was checked using lithium chloride which isknown to block the phosphatidylinositol cycle in animal cells.This compound hindered auxin-induced opening for concentrationshigher than 5x10^–4 M. The calcium-binding protein calmodulinwas shown to be implicated in the IAA-induced response sinceopening was inhibited in a concentration-dependent manner aftertreatment with compound 48/80 and with W-7. Key words: Cassia fasciculata, auxin, calcium, second messenger, turgor regulation     

Respiratory Efflux in Relation to Temperature of Simulated Swards of Perennial Ryegrass with Contrasting Soluble Carbohydrate Contents

Fully light-intercepting simulated swards of S24 perennial ryegrasswere exposed to contrasting environmental conditions in a growthroom for 4 days. Half experienced 20 h days of 120 Wm^–2(400–700. nm) and 5 °C, and came to have a WSC (watersoluble carbohydrate) content of 235 mg g^–1 and half 4h days of 20 Wm^–2 and 25 °C leading to a WSC of 25mg g^–1. Their rates of CO₂ efflux were monitored at anumber of temperatures during an 8 h dark period; half experiencedincreasing (5–30 °C) and half decreasing (30–5°C) temperatures. The ‘high’ WSC swards hadrespiration rates of 3.7 mg CO₂ g^–1 (d. wt) h^–1at 15 °C, and the ‘low’ swards 0.8 mg CO₂ g^–1h^–1. The order in which the temperatures were experiencedwas immaterial. Even the ‘low’ WSC swards showedno evidence of a respiratory decline during the dark periodthat could be attributed to substrate shortage. The relationshipbetween temperature and CO₂ efflux was best represented by logisticcurves. Even so, a Q₁₀ of 2 fitted the data reasonably well,at least up to 20 °C, and has practical advantages wheninterpolating estimated between measured values of respirationin the construction of a carbon balance sheet. Lohum perenne L., ryegrass, respiration, temperature, Q₁₀, soluble carbohydrate content, simulated sward     

Induction of DNA synthesis and callus formation from tuber tissue of Jerusalem artichoke by 2,4-dichlorophenoxyacetic acid

Cellus induction was observed from Jerusalem artichoke tubertissue on a synthetic medium containing 2,4-D at 10^–6,10^–5 (optimum conc.) and 10^–4 M. The first DNA synthesis(thymidine incorporation) was observed only at 2,4-D concentrationsof 10^–5 to 10^–4M. In 10^–5 M 2,4-D treatedtissue, DNA synthesis increased after a 20 hr lag and reacheda maximum at 36 hr, after which it decreased. Actinomycin Dand 8-aza-guanine; inhibitors of RNA synthesis, inhibited DNAsynthesis completely. 2,4-D caused the characteristic changesin RNA and protein syntheses. In comparison with the control,RNA and protein syntheses were first repressed then inducedbefore the peak of DNA synthesis. Treatment with cycloheximide(10^–4M) for one hour before inoculation inhibited proteinsynthesis completely for 12 hr; consequently DNA synthesis wasalso delayed. The results suggest that RNA and protein synthesesneeded for callus induction are regulated by 2,4-D in the firstDNA synthesis. (Received July 19, 1973; )