Control of the germination of secondary dormant cocklebur seeds by various germination stimulants

The responsiveness of non-dormant, upper cocklebur (Xanthiumpennsylvanicum Wallr.) seeds to various germination stimulants,such as CO₂ C₂H₄ CS(NH₂)₂, BA and enriched O₂, decreased withincreasing periods of water imbibition and was completely lostin the state of secondary dormancy. Unlike CO₂ BA and CS(NH₂)₂however, C₂H₂ and enriched O₂ effectively prevented the developmentof secondary dormancy, and their combination was the most effectivefor stimulating the germination of seeds which had undergoneimbibition for a long time. CS(NH₂)₂ and BA were effective,not by themselves but either under anaerobiosis or elevatedO₂ tension. Growth of the axial and cotyledonary segments excisedfrom aged seeds remained responsive to these germination stimulantsand could be further stimulated by exogenous C₂H₂. With imbibitionat a lower temperature, the seeds maintained high germinationin response to various stimulants and a high rate of C₂H₂ andCO₂ production during a long period of water imbibition. Theseresults are discussed in terms of the two possible causes forthe loss of responsiveness or induction of the secondary dormancy. (Received June 27, 1978; )     

不同灌溉处理对铁观音茶树光合作用的影响

以田间栽培的2年生铁观音茶树为试验材料,应用叶绿素荧光诱导动力学技术,以不灌溉为对照,分析不同灌溉间隔时间处理［5 d（T₁）、10 d (T₂)、15 d (T₃)、20 d (T₄) 和25 d (T₅)］对铁观音茶树叶片光合作用的影响.结果表明: 随着灌溉间隔期的延长,2年生铁观音茶树叶片水势和叶绿素含量降低; 净光合速率(P_n)先上升后降低,在T₂下达到最大(15.55 μmol·m^-2·s^-1); 光系统Ⅱ(PSⅡ)的原初光能转化效率(F_v/F_m)、可变荧光衰减(ΔF_v)和可变荧光淬灭速率(ΔF_v/F_o)均在T₂下达到最大值,分别为0.844、342.5和4.03.初始荧光(F_o)随着灌溉间隔期的延长而降低,而对照的F_o则呈上升趋势,表明干旱可对茶树叶片PSⅡ造成损害.灌溉间隔期为10 d处理有利于茶树叶片光合电子的传递和CO₂的同化,提高茶树的光合作用效率.     

Control of cocklebur seed germination by nitrogenous compounds : Nitrite, nitrate, hydroxylamine, thiourea, azide and cyanide

Germination of non-dormant upper cocklebur (Xanthium pinsylvanicumWallr.) seeds was stimulated by not only CS(NH₂)₂ but also NH₂OH,KCN and NaN₃. This stimulation was not via the enhancement ofaerobic C₂H₄ production. NH₂OH, KCN and NaN₃ in certain concentrationspromoted the initial growth of axial and/or cotyledonary parts,but the degree of growth promotion by NH₂OH, NaN₃ and KCN wasslight compared with that by CS(NH₂)₂. As in the case of CS(NH₂)₂,however, the germinationstimulating effect of NH₂OH disappearedrapidly as the preceding imbibition period was prolonged. Incontrast, KCN and NaN₃ were still effective in stimulating thegermination of aged seeds maintained on a water substratum,as previously seen with anaerobiosis. Anaerobic induction wasenhanced not only by NaN₃ and KCN but also by NH₂OH, KNO₃, KNO₂CO(NH₂)₂ and CS(NH₂)₂ applied during the anaerobic treatment,but without causing an increase in anaerobic production of C₂H₄.Furthermore, KCN and NaN₃, given prior to the anaerobic treatmentacted additively with anaerobic induction. The germination-stimulatingactions of nitrogenous compounds are discussed in comparisonwith those of C₂H₄ and anaerobiosis. (Received May 6, 1978; )     

Accumulation of Hydrogen Peroxide in Chloroplasts of SO2-Fumigated Spinach Leaves

Illuminated chloroplasts isolated from SO₂-fumigated spinachleaves accumulated more H₂O₂ than those from non-fumigated ones.This H₂O₂ formation was dependent on light and was inhibitedby DCMU. It also was depressed by cytochrome c and superoxidedismutase (EC 1.15.1.1 [EC] ). The addition of sulfite to rupturedchloroplasts isolated from non-fumigated leaves caused an H₂O₂accumulation that accompanied O₂ uptake. Spinach leaves losttheir catalase (EC 1.11.1.6 [EC] ), ascorbate peroxidase and glutathionereductase (EC 1.6.4.2 [EC] ) activities at the beginning of SO₂ fumigation,when H₂O₂ was accumulated. These results suggest that the accumulationof H₂O₂ in SO₂-fumigated spinach leaves is caused by the increasein O₂^–production, the precursor for H₂O₂, with a sulfite-mediatedchain reaction at the reducing site of photosystem I, and byinactivation of the H₂O₂ scavenging system. (Received October 7, 1981; Accepted June 16, 1982)     

A Modified Micro-Drop Bioassay Using Dwarf Rice for Detection of Femtomol Quantities of Gibberellins

The sensitivity of the micro-drop assay with dwarf rice (Oryzasativa L., cv. Tan-ginbozu and cv. Waito-Q to gibberellins (GAs)was increased conspicuously by the use of assay plants thathas been treated with uniconazole (S-3307), an inhibitor ofthe biosynthesis of GAs. The Tan-ginbozu plants treated withS-3307 responded to 10 fmol/plant of GA₃ (ca. 3.5 pg/plant)and to 30 fmol/plant of gibberellins A₁, A₄, A₇, A₁₉ and A₂₀.Waito-C plants treated with S-3307 responded to 10 fmol of GA₃and to 30 fmol/plant of gibberellins A₁, A₄ and A₇. GibberellinsA₉, A₁₉ and A₂₀ had much less of an effect on the treated Waito-Cplants than did gibberellins A₁, A₃, A₄ and A₇. Furthermore,treatment with S-3307 counteracted the inhibition of growthof both cultivars by abscisic acid. Thus, the modified micro-dropassay should prove very useful for the detection of minute amountsof GAs in plant extracts. (Received October 3, 1988; Accepted March 29, 1989)     

Aqueous two-phase extraction coupled with ultrafiltration for purification of amyloglucosidase

Aqueous two phase extraction (ATPE) in combination with ultrafiltration was employed for concentration and purification of amyloglucosidase produced by solid state fermentation. After extraction (with water) from dry moldy bran the dilute enzyme extract was concentrated by ATPE in a polyethylene glycol (PEG)/maltodextrin (MDX) system. The enzyme in the top PEG rich phase was then extracted into a Na₂HPO₄ rich bottom phase and further concentrated by ultrafiltration. The partitioning behavior of amyloglucosidase was examined in PEG/MDX, PEG/Na₂SO₄, PEG/Na₂HPO₄, PEG/KH₂PO₄ aqueous two phase systems. Effect of buffering salts such as NaCl, Na₂HPO₄, KH₂PO₄ and Na₂SO₄ on the partitioning behavior of enzyme was studied in PEG/MDX system. Maximum partitioning of amyloglucosidase was seen with KH₂PO₄ (m = 18.1). A two stage ATPE employing PEG/MDX (buffered with KH₂PO₄) and PEG/Na₂HPO₄ systems, followed by ultrafiltration has resulted in an overall recovery of 78.4% with 3.1 fold purification and 9.4 fold concentration of the enzyme.     

Partial Reduction in Activities of Photorespiratory Enzymes in C3-C4 Intermediates of Alternanthera and Parthenium

The maximum catalytic activities of several photorespiratoryand photosynthetic enzymes were determined in leaf extractsof three C₃–C₄ intermediates (Alternanthera ficoides,A. tenella and Parthenium hysterophorus) and were compared tothose of C₃ (A. sessiles, Pisum sativum) and C₄ (A. pungens,Zea mays and Amaranthus hypochondriacus) species. The activitylevels of key photorespiratory enzymes, glycolate oxidase, catalase,NADH-hydroxypyruvate reductase and glycerate kinase were less(28 to 35% reduced) in intermediates than those of typical C₃species. Similarly, the activities of photorespiratory aminotransferasesin the C₃–C₄ intermediates were also partially reduced(23 to 37% reduction). The activities of phosphoenolpyruvatecarboxylase (PEPC), pyruvate, orthophosphate dikinase and NAD-malicenzyme were higher (2 to 7 times) in leaf extracts of the intermediatesthan those of C₃ species. But the ratios of PEPC/rubisco inthe C₃–C₄ intermediates were more like C₃ than C₄ species.We draw attention to the partial reduction in enzyme activityof photorespiratory metabolism, which could be an importantfactor for restriction of photorespiration in the C₃–C₄intermediate species, in addition to enzyme compartmentationand/or operation of a ‘C₄-like’ cycle Key words: C₃–C₄ intermediates, C₄ pathway, enzyme profile, glycolate metabolism, photorespiration, photosynthesis     

Identification and Quantification of Cambial Region Hormones of Eucalyptus globulus

Gibberellins GA₁, GA₄, GA₈, GA₉, GA₁₉, GA₂₀, GA₂₉, GA₄₄, GA₈₁,indole-3-acetic acid (IAA) and abscisic acid (ABA) were identifiedin cambial region tissues of Eucalyptus globulus by comparingmass spectra and Kovats retention indices with those of authenticstandards. Using stable isotope labelled internal standardsGA₁₉, GA₂₀ and GA₄₄ were quantified at levels of 2–7 ng(g fr wt)^-1, other GAs were present at levels < 1 ng (g frwt)^-1. Levels of IAA and ABA ranged from 417–1, 140 ng(g fr wt)^-1 and 86–305 ng (g fr wt)^-1 respectively. Thepresence of brassinosteroid-like substances was also indicatedbased on activity in the rice seedling leaf inclination assay. (Received April 28, 1995; Accepted June 20, 1995)     

Fluctuation of Endogenous Levels of Free and Conjugated Gibberellins throughout Seed Maturation in Leucaena leucocephala (Lmk) De Wit

Combined gas chromatography-selected ion monitoring (GC-SIM)analysis of a purified extract from seeds of Leucaena leucocephalashowed the presence of GA₁, GA₈, GA₁₇, GA₁₉, GA₂₀, GA₂₃, GA₂₉and GA₅₃. GA₁, GA₈ and GA₂₉ were also found both as glucosylester-like and glucosyl ether-like conjugates, and GA₂₀ as aglucosyl ester-like conjugate; these conjugates were analyzedas free GAs after enzyme hydrolysis. GA₂₃ was shown to be themain free gibberellin in immature seeds (268 ng/seed), thoughits level drastically decreased during their maturation. GA₁was the main free C₁₉-gibbercllin and GA₁ glucosyl ester-likeand glucosyl ether-like conjugates were found at the highestlevels in the seeds prior to maturation. Fluctuation of endogenouslevels of gibberellins is discussed in terms of seed development. (Received May 9, 1984; Accepted August 25, 1984)     

Analysis by {delta}13C Measurement on Mechanism of Cultivar Difference in Leaf Photosynthesis of Rice (Oryza sativa L.)

Our previous paper showed that cultivar difference of flag leafphotosynthesis (LPS) in rice (Oryza sativa L.) was attributedto the difference in mesophyll resistance (r_m). In this paper,we tried to divide r_m into CO₂ transfer resistance (r_m) andCO₂ fixation resistance (r_c) for further analysis of r_m by meansof     

Osmotic potential of Norway spruce [Picea abies (L.) Karst.] secondary phloem in relation to anatomy

Variations in water status of secondary phloem of Picea abies (L.) Karst., caused by (1) radial and (2) vertical differences within the tree, (3) seasonal influences, and (4) tree class, and their relation to bark anatomy were investigated. The water status parameters measured were the osmotic potential at full saturation [O_{o (sat)}], the in situ osmotic potential [O_{o (in situ)}], the in situ water content (C_{in situ}), and the in situ relative water content (R_{in situ}). O_{o (sat)} reached most negative values in the conducting part of the secondary phloem, whereas O_{o (in situ)} was similar in conducting (P_C) and non-conducting secondary phloem (P_N). The remarkable discontinuity in the radial course of C_{in situ} and O_{o (sat)} at the transition from P_C to P_N can be attributed to the degeneration of sieve cells and Strasburger cells. In P_C, the vertical decrease of O_{o (sat)} towards the crown was compensated by an increase in R_{in situ}, so that O_{o (in situ)} did not change along the stem. With stem height, O_{o (sat)} decreased and P_C width increased. The determining factor for vertical gradients in O_{o (sat)} was the distance to the sources; similar gradients were also measured in P_N. Seasonal differences in O_{o (sat)} could only be detected in P_C, where they corresponded exactly to changes of O_{o (sat)} in needles. Suppressed trees showed less negative O_{o (sat)} values in P_C, smaller annual secondary phloem increments and smaller radial lumen diameter of living sieve cells than predominant or dominant trees.     

Role of H2O2 and heme-containing O2 sensors in hypoxic regulation of tyrosine hydroxylase gene expression

In the currentstudy, we investigated links betweenO₂-regulatedH₂O₂formation and the hypoxic induction of mRNA for tyrosine hydroxylase(TH), the rate-limiting enzyme in catecholamine synthesis, inO₂-sensitive PC-12 cells. Duringexposure of PC-12 cells to 5% O₂,H₂O₂concentration decreased by 40% as measured with2',7'-dichlorofluorescein (DCF). Treatment withH₂O₂reduced TH mRNA during normoxia and prevented the induction of TH mRNAduring hypoxia. Treatment with catalase orN-(2-mercaptopropionyl)-glycine, areducing antioxidant agent that decreasesH₂O₂concentration, also induced TH mRNA. Deferoxamine (DF), an ironchelator, failed to affectH₂O₂formation but induced TH mRNA in normoxia and hypoxia.CoCl₂ led to a decrease inH₂O₂at 20 h of treatment but induced TH mRNA during normoxia and hypoxiabefore it affectedH₂O₂.In conclusion, TH gene expression correlates inversely withH₂O₂formation. DF and Co²⁺ seem toaffect TH gene expression in themechanism downstream from theH₂O₂formation rather than by interfering with theH₂O₂-generating activity of the O₂ sensor.

     

Effects of shell morphological traits on the weight traits of Manila clam (Ruditapes philippinarum)

The shell length, height, and width, live body weight, and edible tissue weight of Manila clam of 1, 2, and 3 years of age were measured, and their correlation coefficients were calculated. The shell morphological traits were used as independent variables, and live body weight or edible tissue weigh used as a dependent variable for calculating the path coefficients, correlation index and determination coefficients. The results showed that the correlation coefficients between each shell morphological trait and the live body weight or edible tissue weight were all highly significant (P < 0. 01). The shell height at 1-year old clams was highly correlated with the live body weight and edible tissue weight. The shell width of 2- to 3-year-old clams was strongly associated with the live body weight, while the shell length was closely linked to the edible tissue weight. The results of coefficients of determination for the morphological traits against weight traits agreed well with the results of path analysis. The correlation indices for all morphological traits against weight traits were approximately the same as determination coefficients regardless of clam age. The correlation indices (R²) of morphological traits against the live body weight of clams of all ages and edible tissue weight of 1-year-old clams were larger than 0.85, but R² of morphological traits against the edible tissue weight of 2- and 3-year-old clams was smaller than 0.85, indicating that some other factors might be associated with the edible tissue weight of 2- and 3-year-old clams. Multiple regression equations were obtained to estimate shell length X₁ (cm), shell height X₂ (cm), shell width X₃ (cm) against live body weight Y (g), edible tissue weight Z (g): for 1-year-old clams: Y = ?4.317 + 0.18X₁ + 0.147X₂, (X₁ < 0.01, X₂ < 0.01), Z = ?1.011 + 0.095X₂, (X₂ < 0.01); for 2-year-old clams: Y = ?15.119 + 0.249X₁ + 0.176X₂ + 0.688X₃, (X₁ < 0.01, X₃ < 0.01), Z = ?4.248 + 0.198X₁, (X₁ < 0.05, X₃ < 0.01); and for 3-year-old clams: Y = ?25.013 + 0.415X₁ + 1.184X₃, (X₁ < 0.01, X₃ < 0.01), Z = ?7.082 + 0.119X₁ + 0.332X₃, (X₁ < 0.05, X₃ < 0.01).     

The Biological Activity of Organoboron Compounds in the Promotion of Root Growth

The growth of the bean (Vicia faba var. minor) radicle in nutrientsolution requires the presence of borate. Optimum extensiongrowth, over 41 h, was obtained in the presence of 0.5 µMboric acid. This requirement of borate for root growth couldalso be satisfied by PhB(OH)₂, 2-OCH₃PhB(OH)₂, and 4-OCH₃PhB(OH)₂.These three compounds also complex in vitro with catechol-3,5-disulphonic acid. NaPh₄B and 2, 6-OCH₃)₂PhB(OH)₂ did not formsuch complexes in vitro, but were biologically active as sourcesof boron for radicle growth. This activity of NaPh₄B was absentif roots were grown in solutions which were changed every 8h. The activity of both NaPh₄B and 2,6-(OCH₃)₂PhB(OH)₂ was increasedby using stock aqueous solutions which were not freshly prepared.Theresults are considered to provide support for the hypothesisthat the activity of borate, as an essential plant nutrient,depends upon its ability to form a biologically active complexwith an in vivo cis-diol compound.     

Oximetry of retinal vessels by dual-wavelength imaging: calibration and influence of pigmentation

A method for noninvasive measurement of HbO₂ saturation(SO₂) inretinal blood vessels by digital imaging was developed and tested.Images of vessels were recorded atO₂-sensitive andO₂-insensitive wavelengths (600 and 569 nm, respectively) by using a modified fundus camera with animage splitter coupled to an 18-bit digital camera. Retinal arterialSO₂ wasvaried experimentally by having subjects breathe mixtures ofO₂ andN₂ while systemic arterial SO₂ wasmonitored with a pulse oximeter. Optical densities (ODs) of vascularsegments were determined using a computer algorithm to track the pathof reflected light intensity along vessels. During graded hypoxia theOD ratio (ODR = OD₆₀₀/OD₅₆₉)bore an inverse linear relationship to systemicSO₂.Compensation for the influence of choroidal pigmentation significantlyreduced variation in the arterialSO₂measurements among subjects. An O₂sensitivity of 0.00504 ± 0.00029 (SE) ODRunits/%SO₂was determined. Retinal venousSO₂ atnormoxia was 55 ± 3.38% (SE). Breathing 100%O₂ increased venousSO₂ by19.2 ± 2.9%. This technique, when combined with bloodflow studies in human subjects, will enable the study of retinalO₂ utilization under experimentaland various disease conditions.     

Role of Carbonic Anhydrase in Photosynthesis of Blue-Green Alga (Cyanobacterium) Anabaena variabilis ATCC 29413

The affinity for NaHCO₃ (CO₂) in photosynthesis of Anabaenavariabilis ATCC 29413 was much higher in the cells grown underordinary air (low-CO₂ cells) than in those grown in air enrichedwith 2–4% CO₂ (high-CO₂ cells) (pH 8.0, 25?C). Ethoxyzolamide(50 µM) increased the K_m(NaHCO₃ in low-CO₂ cells aboutnine times (from 14.3 to 125), while the maximum rate of photosynthesisdecreased about 20%. When high-CO₂ cells were transferred tolow-CO₂ conditions, carbonic anhydrase (CA) activity increased,while K_m(NaHCO₃) in photosynthesis decreased from 140 to 30µM within about 5 h. The addition of CA to the suspensionof both high- and low-CO₂ cells enhanced the rates of photosyntheticO₂ evolution under CO₂-limiting conditions. The rate of ¹⁴CO₂fixation was much faster than that of H¹⁴CO₃^– fixation.The former reaction was greatly suppressed, while the latterwas enhanced by the addition of CA. These results indicate thatthe active species of inorganic carbon utilized for photosynthesiswas free CO₂ irrespective of the CO₂ concentration given duringgrowth. It is suggested that CA plays an active role in increasingthe affinity for CO₂ in photosynthesis of low-CO₂ cells of thisblue-green alga. (Received January 24, 1984; Accepted October 22, 1984)     

Pb2+胁迫对两种小麦幼苗生理特性影响的研究

以两种小麦西旱2号和宁春4号幼苗为供试材料,研究了不同浓度Pb(NO₃)₂处理对其叶绿素含量、抗氧化酶活性及渗透性调节物等生理特性的影响。结果发现,两种小麦在低浓度铅处理下幼苗叶片叶绿素含量及超氧化物歧化酶（SOD）活性均无显著变化,而高浓度铅胁迫使其叶绿素a（Chla）、叶绿素b（Chlb）及叶绿素总量明显减少,但SOD活性显著升高,且相同浓度铅胁迫对小麦宁春4号幼苗叶片叶绿素的破坏作用明显强于对西旱2号的作用;不同浓度硝酸铅处理诱导两种小麦幼苗过氧化氢酶（CAT）、过氧化物酶（POD）和抗坏血酸过氧化物酶（APX）活性升高,但不影响丙二醛（MDA）含量;此外,Pb²⁺处理使小麦幼苗叶片脯氨酸含量升高,此效应具有浓度依赖性,但铅处理不影响可溶性糖相对含量。结果表明,铅胁迫对两种小麦幼苗叶片叶绿素造成了破坏,却不同程度诱导抗氧化酶活性及脯氨酸含量升高,即表现出较强的抗氧化能力和渗透调节能力,增强了小麦对铅的耐受性,因而胁迫诱导两种小麦叶片MDA含量变化与对照比无显著性差异。     

Plasmalemma Transport of HCO-3 and OH- in Chara corallina: Inhibitory Effect of Ammonium Sulphate

The influence of (NH₄)₂SO₄ on ¹⁴C assimilation and cyclosisin internodal cells of Chara corallina was investigated. Severeinhibition of ¹⁴C assimilation was found at pH values above7·0, this inhibition being correlated with the exogenouslevel of NH3 rather than NH⁺₄. Cyclosis was also affected athigher concentrations of (NH₄)₂SO₄. This effect was similarlycorrelated with exogenous levels of NH₃. ¹⁴C assimilation was inhibited non-competitively by (NH₄)₂SO₄,the apparent Km being increased from 0·55 to 1·5mM. The results suggest that the site(s) of inhibition is locatedat the plasmalemma, rather than at the chloroplasts. (Evidencein support of in vivo uncoupling of photophosphorylation, bylow concentrations of (NH₄)₂SO₄, was not obtained). Significant perturbation of the OH^– efflux pattern wasobserved as the level of (NH₄)₂SO₄ was increased. Induced migrationof efflux sites indicates that NH₃ may interfere with the cellularmechanism that controls OH^– transport. Using a cell-segmentisolating chamber it was shown that (NH₄)₂SO₄ inhibited OH^–efflux rather than HCO^–₃ transport. This inhibitory effectwas readily reversible. These data are discussed in terms of a possible relationshipbetween the observe NH₄)²SO₄ stimulation of ³⁶Cl^– influxand the effect of this compound on ¹⁴C assimilation.     

Germination and Storage of Pollen of Phytolacca dodecandra L. (endod)

The effect of sucrose, H₂BO₃, KNO₃, Ca(NO₂)₂.4H₂O and MgSO₄.7H₂O on pollen germination of Phytolacca dodecandra L. (endod)in a liquid medium was investigated. Sucrose and H₃BO₃ werecritical to pollen germination. A concentration of 10% sucroseand 161.8 µm H₂BO₃ gave over 70% germination. The germinationof pollen was not enhanced by Ca(NO₃)₂.4H₂O, KNO₃ and MgSO₄.7H₂O.Endod pollen was dehydrated over CaCl₂ and stored in gelatincapsules in cryogenic vials at –175 °C, 1±1°C and 24±2 °C. The pollen moisture content atcollection was approx. 7.8% (f. wt basis) and dehydration overCaCl₂ reduced it to about 1.4%. Pollen stored at 1±1°C and –175 °C maintained viability for over 6months. Pollen stored at room temperature lost viability within4 weeks of storage. Pollination with cryopreserved pollen resultedin normal fruit set. Phytolacca dodecandra, endod, pollen germination, pollen storage     

Phase Shifting Effects in the Photoperiodic Rhythm of Flowering in Dark-Grown Seedlings of Pharbitis nil Choisy

Dark-grown seedlings of Pharbitis nil Choisy received an initialsaturating fluence of red (R) light (R₁), followed at intervalsby further R pulses (R₂ and R₃). R₂ was given at different timesafter R₁. R₂ was used to scan the subsequent 72 h period. The initial exposure to R (R₁) initiated a circadian rhythmin the flowering response to the scanning R exposure (R₂). Thephase of the rhythm was shifted by the second exposure to R(R₂) and the sensitivity of the phase-shifting response variedwith the time of giving the R₂ pulse. The direct response toR₂ (i.e., the magnitude of flowering produced in the absenceof a scanning R₂ exposure) also varied in sensitivity. WhenR² was given 4h after R₁, the phase-shift was achieved by anexposure of 20 s (sufficient to establish 20–25% Pfr/P)but more than 80 s was required to saturate the direct floweringresponse at this time. When given 16 h after R₁, 80 s of R₂(sufficient to establish 55% Pfr/P) was required for the phase-shift,whereas the maximum promotion of flowering was produced by only5 s R. These differences in fluence-response relationships indicatethat the direct flowering response to a dark interruption withR and the effect of such an interruption to phase-shift theunderlying rhythm are distinct processes. (Received April 30, 1986; Accepted November 11, 1986)