Betaine Improves Freezing Tolerance in Wheat

The accumulation of the osmolyte betaine was found to be correlatedwith the development of freezing tolerance (FT) of two wheatcultivars where it increases by about three fold during thecold acclimation period. Exogenous betaine application resultedin a large increase in total osmolality mostly due to betaineaccumulation. Plants that accumulated betaine are more tolerantto freezing stress since a four day exposure to 250 mM betaineresulted in a LT₅₀ of –8°C (in spring wheat Glenlea)and –9°C (in winter wheat Fredrick) compared to –3°C(Glenlea) and –4°C (Fredrick) for control non-exposedplants. Betaine treatment (250 mM) during cold acclimation increasedFT in an additive manner since the LT₅₀ reached –14°C(Glenlea) and –22°C (Fredrick) compared to –8°C(Glenlea) and –16°C (Fredrick) for plants that arecold acclimated in the absence of betaine. These results showthat betaine treatment can improve FT by more than 5°C inboth non-acclimated and cold-acclimated plants. The betainetreatment resulted in the induction of a subset of low temperatureresponsive genes, such as the wcor410, and wcor413, that arealso induced by salinity or drought stresses. In addition tothese genetic responses, betaine treatment was also able toimprove the tolerance to photoin-hibition of PSII and the steady-stateyield of electron transport over PSII in a manner that mimickedcold-acclimated plants. These data also suggest that betaineimproves FT by eliciting some of the genetic and physiologicalresponses associated with cold acclimation. (Received April 23, 1998; Accepted September 4, 1998)     

Changes in the Translatable RNA Population during Abscisic Acid Induce Freezing Tolerance in Bromegrass Suspension Culture

Abscisic acid (ABA) has been shown to increase freezing toleranceof bromegrass (Bromus in-ermis Leyss cv. Manchar) cell suspensioncultures from a LT₅₀ (the temperature at which 50% cells werekilled) of –7 to – 30?C in 5 days at 23?C. Our objectivewas to study the qualitative changes in the translatable RNApopulation during ABA induced frost tolernace. In vitro translationproducts of poly(A)⁺ RNA isolated from bromegrass cells withor without 75 µM ABA treatment for various periods oftime were separated by 2D-PAGE and visualized by fluorography.SDS soluble proteins from the same treatments were also separatedby 20-PAGE. After 5 days treatment, at least 22 new or increasedabundance SDS soluble polypeptides were observed. From fluorographs,29 novel or increased abundance in vitro translation productscould be detected. The pattern of changes between ABA inducedSDS-soluble proteins and translation products from the 2D gelswere similar. A time course study (0–7 days) showed that17 of the 29 translation products were detected after 1 dayABA treatment, and at least 14 were present after 1 h. Coldtreatment (+4?C) induced fewer changes in the pool of translatableRNA than with ABA treatment. Three translation products inducedby cold appear to be similar to 3 of the ABA induced translationproducts. The majority of the ABA inducible translatable RNAsappeared at 10 µM or higher which coincides with the inductionof freezing tolerance. Many of these ABA inducible RNAs persisted7 days after ABA was removed from the media and correspondinglythe LT₅₀ (–17?C) was still well above the control level(–17?C). The results suggest that ABA alters the poolof translatable RNAs during induction of freezing tolerancein bromegrass suspension culture cells. ¹Oregon Agricultural Experiment Station Technical Paper No.9256. (Received August 3, 1990; Accepted October 18, 1990)     

Effect of Photoperiod and Irradiance Changes Upon Development of Freezing Tolerance and Accumulation of Soluble Carbohydrate in Seedlings of Lolium perenne Grown at 2 {degrees}C

Two contrasting cultivars of Lolium perenne were exposed toa range of daily radiation integrals during hardening at 2°Cfor 15 d. The maximum induced freezing tolerance measured asLT₅₀ (temperature for 50 % kill) differed markedly between thecultivars. The observed LT₅₀ values were unaffected by changesin the radiation integral above 10 mol m^–2 d^–1,whereas accumulation of water-soluble carbohydrate showed astrong positive correlation with the radiation integral overthe entire range of the experiment. The correlation betweenLT₅₀ and soluble carbohydrate content at the end of the hardeningperiod was poor and showed no obvious connection with genotype.Fructan polymers and sucrose were the major components of thesoluble carbohydrates in both cultivars. The depression of freezingpoint attributable to the accumulation of soluble, osmoticallyactive carbohydrate was not sufficient to account for the observedchanges in LT₅₀ in the hardy genotype. These results are discussedin relation to the interactions between growth, photosynthesisand assimilate partitioning during hardening. Lolium perenne, hardening, freezing tolerance, irradiance, carbohydrate, fructan     

Freezing tolerance, protein composition, and abscisic acid localization and content of pea epicotyl, shoot, and root tissue 3

The freezing tolerance of many plants, such as pea (Pisum sativum),is increased by exposure to low temperature or abscisic acidtreatment, although the physiological basis of this phenomenonis poorly understood. The freezing tolerance of pea shoot tips,root tips, and epicotyl tissue was tested after cold acclimationat 2C, dehydration/rehydration, applications of 10^–4M abscisic acid (ABA), and deacclimation at 25C. Tests wereconducted using the cultivar ‘Alaska’, an ABA-deficientmutant ‘wil’, and its ‘wildtype’. Freezinginjury was determined graphically as the temperature that caused50% injury (T₅₀) from electrical conductivity. Endogenous ABAwas measured using an indirect enzyme-linked immunosorbant assay,and novel proteins were detected using 2-dimensional polyacrylamidegel electrophoresis. The maximum decrease in T₅₀ for root tissuewas 1C for all genotypes, regardless of treatment. For ‘Alaska’shoot tips and epicotyl tissue, exogenous ABA increased thefreezing tolerance by –1.5 to –4.0C, while coldtreatment increased the freezing tolerance by –7.5 to–14.8C. Cold treatment increased the freezing toleranceof shoot tips by –9 and –15C for ‘wil’and ‘wild-type’, respectively. Cold acclimationincreased endogenous ABA concentrations in ‘Alaska’shoot tips and epicotyls 3- to 4-fold. Immunogold labeling increasednoticeably in the nucleus and cytoplasm of the epicotyl after7 d at 2C and was greatest after 30 d at the time of maximumfreezing tolerance and soluble ABA concentration. Cold treatmentinduced the production of seven, three, and two proteins inshoot, epicotyl, and root tissue of ‘Alaska’, respectively.In ‘Alaska’ shoot tissue, five out of seven novelproteins accumulated in response to both ABA and cold treatment.However, only a 24 kDa protein was produced in ‘wil’and ‘wild-type’ shoot and epicotyl tissues aftercold treatment. Abscisic acid and cold treatment additivelyincreased the freezing tolerance of pea epicotyl and shoot tissuesthrough apparently independent mechanisms that both resultedin the production of a 24 kDa protein. Key words: Pisum sativum, cold acclimation, immuno-localization     

Developmental control of xylem hydraulic resistances and vulnerability to embolism in Fraxinus excelsior L.: impacts on water relations

The freezing tolerance of many plants, such as pea (Pisum sativum),is increased by exposure to low temperature or abscisic acidtreatment, although the physiological basis of this phenomenonis poorly understood. The freezing tolerance of pea shoot tips,root tips, and epicotyl tissue was tested after cold acclimationat 2C, dehydration/rehydration, applications of 10^–4M abscisic acid (ABA), and deacclimation at 25C. Tests wereconducted using the cultivar ‘Alaska’, an ABA-deficientmutant ‘wil’, and its ‘wildtype’. Freezinginjury was determined graphically as the temperature that caused50% injury (T₅₀) from electrical conductivity. Endogenous ABAwas measured using an indirect enzyme-linked immunosorbant assay,and novel proteins were detected using 2-dimensional polyacrylamidegel electrophoresis. The maximum decrease in T₅₀ for root tissuewas 1C for all genotypes, regardless of treatment. For ‘Alaska’shoot tips and epicotyl tissue, exogenous ABA increased thefreezing tolerance by –1.5 to –4.0C, while coldtreatment increased the freezing tolerance by –7.5 to–14.8C. Cold treatment increased the freezing toleranceof shoot tips by –9 and –15C for ‘wil’and ‘wild-type’, respectively. Cold acclimationincreased endogenous ABA concentrations in ‘Alaska’shoot tips and epicotyls 3- to 4-fold. Immunogold labeling increasednoticeably in the nucleus and cytoplasm of the epicotyl after7 d at 2C and was greatest after 30 d at the time of maximumfreezing tolerance and soluble ABA concentration. Cold treatmentinduced the production of seven, three, and two proteins inshoot, epicotyl, and root tissue of ‘Alaska’, respectively.In ‘Alaska’ shoot tissue, five out of seven novelproteins accumulated in response to both ABA and cold treatment.However, only a 24 kDa protein was produced in ‘wil’and ‘wild-type’ shoot and epicotyl tissues aftercold treatment. Abscisic acid and cold treatment additivelyincreased the freezing tolerance of pea epicotyl and shoot tissuesthrough apparently independent mechanisms that both resultedin the production of a 24 kDa protein. Key words: Pisum sativum, cold acclimation, immuno-localization     

Evidence for the Cell Wall Involvement in Temporal Changes in Freezing Tolerance of Jerusalem Artichoke {Helianthus tuberosus L.) Tubers during Cold Acclimation

We studied the mechanism of cold acclimation of Jerusalem artichoke{Helianthus tuberosus L.) tubers with special reference to therole of the cell wall. During the cold-acclimation process fromSeptember to January, the freezing tolerance of tubers increasedfrom – 2.8°C to –8.4°C (LT₅₀). By contrast,the isolated protoplasts con- stitutively showed a consistenthigh level of freezing toler ance (LT₅₀; below – 25°C)throughout the period. In tuber tissues, freezing injury waseffectively protected by the ex ternal addition of isotonicsolutions. Cryomicroscopic ob servations revealed that tissuecells mounted in isotonic so lutions plasmolyzed upon freezing;tissue cells mounted in water collapsed with a tight attachmentof plasma mem brane to the cell wall. Upon freezing of intacttissues in water to temperatures below the critical range, thecyto plasm was irreversibly acidified as revealed by a fluorescence pH-ratiometry, suggesting that occurrence of detri mentalcellular events leading to permanent cell injury. The freeze-inducedacidification of cytoplasm was also effective ly prevented bythe external addition of isotonic solutions. These results suggestthat the tight attachment of the plas ma membrane to the cellwall during freezing may have a harmful effect on cells, inparticular on the plasma mem brane, possibly due to mechanicalor some sort of chemi cal/physico-chemical interaction withthe cell wall. ¹Contribution no. 3946 from The Institute of Low TemperatureScience, Hokkaido University. This research was supported inpart by the grant from Japan Society for the Promotion of Science(JSPS-RFTF 96L00602) ²Present address: Tohoku National Agricultural Experiment Station, Morioka, Iwate, 020-01 Japan     

Early Developments in RNA, Protein, and Sugar Levels during Cold Stress in Winter Rye (Secale cereale) Leaves

Changes in freezing tolerance of winter rye (Secale cerealeL. cv. Voima) were determined for leaf tissues during a 1-weekcold stress, which was performed by transferring the 7-d-oldseedlings from a greenhouse (25°C, long day) to 3°Cand short day conditions. The development of cold hardeningwas shown by using an ion leakage test and by determining theamounts of carbohydrates, soluble proteins and RNA. The firstevidence of the development of freezing resistance was foundafter 1 d at low temperature, i.e. an LT₅₀ value increased from-5 to -7°C. Plants cold treated for 7 d reached an LT₅₀value of -9°C. This increase in freezing tolerance was foundto be associated with the increased levels of soluble carbohydrates,total RNA and soluble proteins. These metabolic changes indicatethe association with adjustment of growth and cell metabolismto low temperatures at the beginning of cold acclimation ofwinter rye.Copyright 1994, 1999 Academic Press Secale cereale L., winter rye, cold stress, proteins, RNA, sugars     

Normalizing Development of Cultured Triticum aestivum L. Embryos: I. LOW OXYGEN TENSIONS AND EXOGENOUS ABA

Wheat (Triticum aestivum L.) embryos form in dynamically-regulatedovular environments. Our objectives were to improve developmentof cultured immature wheat embryos by simulating, in vitro,abscisic acid (ABA) levels and O₂ tensions as found in wheatovules during zygotic embryogenesis. We characterized from intactwheat kernels embryo respiration, embryo morphology and embryoand endosperm + ABA levels at 13, 19 and 25 d post-anthesis(DPA). Young (13 DPA) embryos were then excised and culturedin vitro, where they were exposed to 0·2 or 2·Ommol m^–3 ±ABA and 2.·1, 2·5 or 7·4mol m^–3 (6, 7 and 21%, respectively) gaseous O₂. At 6and 12 d in culture, + ABA levels, embryo respiration and embryomorphology were characterized by treatment. Thirteen-day-oldembryos from two different plant populations differed by 17-foldin initial ABA content. However, this difference did not affectprecocious germination in vitro, nor did it affect the amountof exogenous ABA required to reduce precocious germination by40%. In this respect, embryos from both populations were equallysensitive to exogenous ABA. Cavity sap O₂ levels (2·1to 2·5 mol m^–3) were much more effective in preventingprecocious germination of cultured embryos than were cavitysap levels of ABA (0·2 to 2·0 mmol m^–3).The combination of physiological levels of both ABA and O₂ largelynormalized DW accumulation and embryo morphology without alteringendogenous + ABA levels. Residual respiration of cultured embryoswas higher than that of embryos grown in situ, and was not influencedby the exogenous O₂ and ABA treatments Key words: Abscisic acid, embryo development, oxygen tensions, respiration, wheat     

Freezing Tolerance and Solute Changes in Contrasting Genotypes of Lolium perenne L. Acclimated to Cold and Drought

Clonal ramets of 12 contrasting genotypes of Lolium perenneL. were grown in sand or soil-based compost and maintained underwell-watered conditions at 20/15°C or acclimated to lowtemperature (2°C) or to a restricted water supply. Freezingtolerance was measured as LT₅₀ following exposure to sub-zerotemperatures in a freezing tank. Measurements were also madeof osmotic potential, water-soluble carbohydrates, free proline,free amino acids, and minerals in entire tillers. Acclimationto both drought and cold lowered LT₅₀, induced osmotic adjustment,and increased concentrations of proline and amino acids, Rootingmedium had little effect on LT₅₀, but caused large differencesin osmotic potential and in proline and amino-acid concentrations.There was considerable genetic diversity for all charactersmeasured, except for mineral contents. There was, however, norelationship between LT₅₀ and osmotic potential or solute contentthat was consistent across the three sources of variation (growingmedium, acclimation, genotype). Furthermore, the diverse genotypicvalues of cold-induced freezing tolerance were not correlatedwith those of drought-induced tolerance. It is concluded thatmore precise measurements are needed of the partitioning ofsolutes during acclimation and of the sensitivity of differentorgans and tissues to freezing.Copyright 1993, 1999 AcademicPress Perennial ryegrass, hardening, acclimation, osmotic potential, solute potential, carbohydrates, proline     

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     

Concurrent Rates of N2 Fixation, Nitrate and Ammonium Uptake by White Clover in Response to Growth at Different Root Temperatures

Nodulated white clover plants (Trifolium repens L. cv. Huia)were grown for 71 d in flowing nutrient solutions containingN as 10 mmol m_–3 NH₄NO₃, under artificial illumination,with shoots at 20/15°C day/night temperatures and root temperaturereduced decrementally from 20 to 5°C. Root temperatureswere then changed to 3, 7, 9, 11, 13, 17 or 25°C, and theacquisition of N by N₂ fixation, NH₄+ and NO₃^– uptakewas measured over 14 d. Shoot specific growth rates (d. wt)doubled with increasing temperature between 7 and 17°C,whilst root specific growth rates showed little response; shoot:root ratios increased with root temperature, and over time at11°C. Net uptake of total N per plant (N₂ fixation + NH₄++ NO₃^–) over 14 d increased three-fold between 3 and 17°C.The proportion contributed by N₂ fixation decreased with increasingtemperature from 51% at 5°C to 18% at 25°C. Uptake ofNH₄+ as a proportion of NH₄+ + NO₃^– uptake over 14 d variedlittle (55–62%) with root temperature between 3 and 25°C,although it increased with time at most temperatures. Mean ratesof total N uptake per unit shoot f. wt over 14 d changed littlebetween 9 and 25°C, but decreased progressively with temperaturebelow 9°C, due to the decline in the rates of NH₄+ and NO₃^–uptake, even though N₂ fixation increased. The results suggestthat N₂ fixation in the presence of sustained low concentrationsof NH₄+ and NO₄^– is less sensitive to low root temperaturethan are either NH₄+ or NO₃^– uptake systems. White clover, Trifolium repens L. cv. Huia, root temperature, nitrogen fixation, ammonium, nitrate     

Variations in Cold Resistance among Apple Cultivars during Deacclimation

Coleman, W. K. 1985. Variations in cold resistance among applecultivars during deacclimation.——J. exp. Bot. 36:1159–1171. One-year-old vegetative twig samples from mature, bearing treesof nine apple cultivars were monitored over two years for theirdormancy intensity and relative cold hardiness levels duringthe winter/spring deacclimation period. The apple cultivarsexhibited a consistent response during the dehardening processwhich included a higher initiation temperature for the low temperatureexotherm (LT₂) and the development of an intermediate freezingexotherm (LT₁.). Imperial Red Mac/Antonovka was the hardiestcultivar during the two-year period while Imperial Red Mac/M.111was the most tender. Cortland/Beautiful Arcade and Rogers RedMac/M.111 varied considerably in their relative hardiness responsesfrom year to year. Mid-winter hardiness levels were significantlyand positively correlated with dormancy intensity in the ninecultivars. However, this relationship did not exist when thehardiness indices for late winter or early spring were comparedwith dormancy intensity. An intensive correlation and path analysisof the response of four cultivars (Jersey Mac/M.111, Vista Bella/M.111,Spur Mac/M.111 and Rogers Red Mac/M.111) to previous maximum/minimumair temperatures indicated that past maximum temperature primarilyaffected LT₂ while past minimum temperature affected LT₁. Whenlinear regression equations were fitted to the data, the meanair temperature of 0°C coincided with LT₁ values of —18 °C and LT₂ values of –36°C to –38°Cfor all four cultivars. Correlation analyses between % moisturecontent and LT₁/LT₂ for the four cultivars were often positivebut generally non-significant. Injury in living cells slightlypreceded the initiation temperature of LT₁ and supports theidea that membrane destabilization may be an important and immediateprecursor to intracellular freezing. Key words: Apple, cold hardiness, deacclimation     

Effect of Root Zone Temperature on the Mineral Composition of Xylem Sap and Plasma Membrane K+-Mg++-ATPase Activity of Grafted-Cucumber and -Figleaf Gourd Root Systems

Cucumber (Cucumis sativus L.) seedlings were grafted onto cucumber-(CG) or figleaf gourd- (FG, Cucurbita ficifolia Bouché)seedlings in order to determine the effect of solution temperature(12, 22, and 32°C) on the mineral composition of xylem sapand the plasma membrane K⁺-Mg⁺⁺-ATPase activities of the roots.Low solution temperature (12°C) lowered the concentrationof NO^–₃ and H₂PO^–₄ in xylem sap of CG plants butnot of FG plants. Concentrations of K⁺, Ca⁺⁺ and Mg⁺⁺ in xylemsap were less affected than anions by solution temperature.The plasma membrane of FG plants grown in 12°C solutiontemperature showed the highest K⁺- Mg⁺⁺-ATPase activity at allATP concentrations up to 3 mM and at low reaction temperatureup to 12°C, indicating resistance of figleaf gourd to lowroot temperature. (Received December 27, 1994; Accepted March 10, 1995)     

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     

Effect of Light Intensity, Carbon Dioxide Concentration, and Leaf Temperature on Gas Exchange of Spray Carnation Plants

The rates of CO₂ assimilation by potted spray carnation plants(cv. Cerise Royalette) were determined over a wide range oflight intensities (45–450 W m^–2 PAR), CO₂ concentrations(200–3100 vpm), and leaf temperatures (5–35 °C).Assimilation rates varied with these factors in a way similarto the response of single leaves of other temperate crops, althoughthe absolute values were lower. The optimal temperature forCO₂ assimilation was between 5 and 10 °C at 45 W m^–2PAR but it increased progressively with increasing light intensityand CO₂ concentration up to 27 °C at 450 W m^–2 PARand 3100 vpm CO₂ as expressed by the equation TOpt = –6.47-h 2.336 In G + 0.031951 where C is CO₂ concentration in vpmand I is photo-synthetically active radiation in W m^–2.CO₂ enrichment also increased stomatal resistance, especiallyat high light intensities. The influence of these results on optimalization of temperaturesand CO₂ concentrations for carnation crops subjected to dailylight variation, and the discrepancy between optimal temperaturesfor growth and net photosynthesis, are discussed briefly     

ABA Levels and Effects in Chilled and Hardened Phaseolus vulgaris

Leaf abscisic acid (ABA) levels of chilled P. vulgaris weremeasured after 18 h chilling at 5°C, at a saturation deficitof 1.24 g m^–3 (SD), and after chilling in a water-saturatedatmosphere. Changes were also followed during a chill hardeningperiod of 4 d at 12°C, 2.1 g m^–3 SD. It was foundthat hardening resulted in an almost 5. fold increase in ABAlevels after 3 d at 12°C, and this decreased to approximatelycontrol levels on the fourth day. Subsequent chilling of hardenedplants produced no change in ABA levels from that of controlplants (22° C). In contrast, non-hardened plants chilledat 1.24 g m^–3 SD had ABA levels almost 3 times the levelof control plants. However, chilling in a water-saturated atmosphereresulted in a decrease in ABA levels. In addition, the response of leaf diffusion resistance (LDR)to exogenous ABA fed via the transpiration stream was measuredat 5 ° C and 22° C in hardened and non-hardened plants.Use of tritium-labelled ABA was made to calculate the stomatalsensitivity to ABA. It was found that exogenous ABA caused anincreased in LDR at 22°C in both hardened and non-hardenedplants. However, the sensitivity of the hardened plants to ABAwas greater in terms of rate of closure and amount of ABA requiredto close the stomata. At 5°C, however, ABA caused stomatalopening and the maintainance of open stomata in non-hardenedplants. In hardened plants, ABA caused stomatal closure at 5°C.These results are discussed in relation to the locking-openresponse of chilled P. vulgaris stomata. Key words: Chilling, Stomata, ABA, Phaseolus vulgaris     

Effects of growth temperature on photosynthetic carbon metabolism in green plants III. Differences in structure, photosynthetic activities and activities of ribulose diphosphate carboxylase and glycolate oxidase in leaves of wheat grown under varied temperatures

The rate of ferricyanide photoreduction in broken chloroplastsisolated from leaves of wheat acclimatized to a low temperature(mean temperature, 5–7?C) was similar to that in chloroplastsfrom wheat acclimatized to a high temperature (20–25?C). There was no practical difference in glycolate oxidase activityin leaf extracts of wheat plants grown at low and high temperatures.In contrast, the ribulose diphosphate carboxylase activity ofchloroplasts from low temperature sample was less than halfthat for the high temperature sample. Chloroplasts having a high rate of photosynthetic CO₂-fixationwere obtained from wheat acclimatized to a low temperature,whereas the CO₂-fixation activity in chloroplasts isolated fromhigh temperature-acclimatized wheat was very low. Electron microscopy revealed that chloroplasts in high temperature-acclimatizedwheat were ellipsoidal, electron dense and contained starchgranules. Those in low temperature-acclimatized leaves wereround and did not contain starch granule. ²Present address: Department of Botany, Faculty of Science,University of Tokyo, Tokyo, Japan (Received August 7, 1973; )     

Carbon Dioxide Exchange of Spring-wheat in Relation to Age and Photon-flux Density at Different Growth Temperatures

CO₂-exchange rates (CER) of the sixth and the flag leaves oftwo spring-wheat varieties, Kolibri and Famos, were comparedusing an open-circuit infrared gas analysing system. Measurementswere repeated every two weeks starting when leaf blades werefully expanded. Single plants were grown in a controlled environmenthaving a photopuiod of 15 h and a day/night temperature of 24/19°C(H), 18/13 °C (M), and 12/7 °C (L) respectively untilapprox. 2 weeks after anthesis and at 18/13 °C until maturity.The photosynthetic photon-flux density (PPFD) at the top ofthe plants was 500 µE m^–2 sec^–1. During themeasurements PPFD was gradually reduced from 2000 to 0 µEm^–2 sec^–1 whereas the temperature was maintainedat the respctive growth-temperatures during the light period.The CER of the sixth leaf declined fairly similarly for bothvarieties, except for Kolibri where a faster decline was observedduring the first two weeks after full leaf expansion. The CERof the flag leaf declined more slowly than that of the sixthleaf. With the flag leaf of Famos, the decline was nearly linear,whereas with Kolibri it was very slow during the first few weeksbut rapid as the leaves further senesced. This pattern becamemore pronounced as the growth temperature decreased. The declinein relation to leaf age was much smaller at low PPFD than athigh PPFD during the same period. At full leaf expansion Kolibrireached higher maximum CER than Famos except at H. As the PPFDwas reduced the difference became smaller and at very low PPFDsuch as 50 µE m^–2 sec^–1 was reversed for thesixth leaf. Under optimum growth conditions maximum values ofCER were greater than 50mg CO₂ dm^–2h^–1 and PPFDfor light saturation was close to 2000 µE m^–2 sec^–1.A comparison between the actual CER and a fitted curve widelyused, PN=(a+b/l)^–1–DR, showed that the goodnessof fit strongly depends on cultivar, treatment and leaf ageas well as on the number and the level of PPFD from which datafor calculations are taken. Triticum aestivum, L., wheat, photosynthesis, photon-flux density, light response, carbon, dioxide exchange     

The Effects of Temperature and Form of Nitrogen Supply on the Relative Contribution of Root Uptake and Remobilization in Supplying Nitrogen for Laminae Regrowth of Lolium perenne L.

Plants of Lolium perenne L. cv. S23 were grown in sand culturesupplied with either ammonium (NH₄⁺) or nitrate (NO₃^–)in an otherwise complete nutrient solution at 12°C or 20°C.Three weeks after germination, plants were clipped weekly tosimulate grazing. After 10 weeks growth all nitrogen (N) wassupplied enriched with ¹⁵N to quantify the effects of form ofN supply and temperature on the relative ability of currentroot uptake and remobilization to supply N for laminae regrowth. The form of N supply had no effect on the dry matter partitioning,while at 20°C more dry weight was allocated to laminae regrowthand less to the remaining plant material. The current root uptakeof N, which subsequently appeared in the laminae regrowth, wassimilar for plants supplied with NH₄⁺ or NO₃^–, and bothwere equally reduced at the lower temperature of growth. Remobilizationof N to laminae regrowth was greater for plants receiving NH₄⁺than NO₃^–; remobilization with either form of N supplywas reduced at the lower temperature of growth. Remobilizationwas reduced to a lesser extent at 12°C than current rootuptake. It was concluded that remobilization became relativelymore important in supplying N for regrowth of laminae at lowertemperatures. Key words: Lolium perenne, ammonium, nitrate, temperature, remobilization     

Kinetics of Leaf Growth in Lolium temulentum at Optimal and Chilling Temperatures

Lolium temulentum plants were grown at 20 °C, under an 8-hdaylength, in a controlled-environment chamber, and the kineticsof leaf expansion were observed by measuring the movement ofan optical grid attached to the fourth leaf. The leaf emerged23–24 d after sowing and was fully expanded 9–10d later. Extension rate was maximal between the second and fifthdays after emergence and declined markedly thereafter. Duringthe rapid growth phase the rate of elongation exhibited a distinctdiurnal rhythm, fluctuating between 1.9 to 2.3 mm h^–1in the light period, and 1.3 to 1.7 mm h^–1 in the dark.A circadian oscillation with a period of about 27 h was observedin leaves elongating in continuous darkness. When plants weretransferred to 5 °C soon after emergence of the fourth leafthere was an immediate reduction in rate of growth to about22 per cent of the rate at 20 °C: the Q₁₀ for the mean elongationrate in the range 20–5 °C was 3.7. When plants weretransferred from 20 to 2 °C at fourth leaf emergence, meanextension rate declined to less than 5 per cent, correspondingto a Q₁₀ in the range 5–2 °C of more than 300. Furthermore,growth at 2 °C was confined almost entirely to the darkphase of the photoperiod cycle. The responsive tissue was shownto be a small area of expanding leafless than 1.5 cm above theshoot apex and the possible mechanisms underlying low temperatureeffects in this region are discussed. Lolium temulentum L., leaf growth, auxanometer, low temperature, diurnal rhythm