Light- and Auxin-Induced Leaflet Opening in Detached Pinnae of Mimosa pudica

Leaflet pairs from detached pinnae of Mimosa pudica opened afterthe pinnae had been irradiated with light (2 W·m^–2)of 726 or 403 nm, whereas they remained almost closed with lightof 585 or 656 nm. Light-induced leaflet opening was observedonly in the daytime, from 6:00 to 16:00. Application of IAAat more than 30 µg/ml to the cut end of the pinna rachisesmade the leaflets open even in darkness with almost constantlag times of about 100 min which were independent of the concentration.NAA and 2,4-D also made the leaflet open at lower concentrationsthan IAA. Auxin-induced leaflet opening showed diurnal variation.Application of IAA for 2 to 6 min, depending on the concentration,was enough to open the leaflets. Autoradiography showed thatIAA was transferred from the cut end of a rachis throughouta pinna within 4 min. ¹Present address: Biological Institute, Faculty of Science,Kobe University, Kobe 657, Japan. (Received September 24, 1982; Accepted March 4, 1983)     

Photosynthetic Oxygen Production Facilitates Translocation of 11C-Labelled Photoassimilates from Tendrils and Leaflets of Pisum sativum L

The translocation profiles of ¹¹C-photoassimilates from eithertendrils or leaflets of the compound leaf of Pisum sativum weresimilar in shape, speed and susceptibility to blockage by chillingand heat girdling. When the feed leaf component was exposedto an anaerobic gas stream consisting of N₂ gas supplementedwith 40 Pa CO₂, the export of previously-fixed ¹¹C-photoassimilatesfrom both leaflets and tendrils continued in the light, butstopped in the dark. However, in the light, translocation of¹¹C-assimilates from the leaflet was rapidly blocked by a flowof pure N₂ (i.e. anoxia). Movement of ¹¹C-assimilates from theleaf of another C₃ plant, sunflower, was similar to that fromthe pea leaflet. In contrast to both laminar leaf components,export from the tendrils was stopped under pure N₂ only in thedark. Taken together the data suggest that photosynthetic O₂production facilitated the movement of ¹¹C-assimilates in theabsence of exogenous O₂. The differences observed between thetendrils and the leaflets exposed to pure N₂ could be attributedto the greater capacity of tendrils to produce and recycle CO₂to support photosynthetic O₂ production in the light. Key words: Pea, ¹¹C-translocation, anoxia, tendril, leaflet     

Role of pulvinar chloroplasts in light-driven leaf movements of the trifoliate leaf of bean (Phaseolus vulgaris L.)

Laminar pulvini of bean (Phaseolus vulgaris L.) contain numerouschloroplasts in cells of their motor tissue. The quantitativerelationships of the chloroplast pigments, chlorophyll a andb, ß-carotene, lutein, neoxanthin as well as the xanthophyllcycle carotenoids (violaxanthin, antheraxanthin and zeaxanthin)were similar to those of mesophyll chloroplasts from leafletlaminae. Exposure of pulvinules to light caused deepoxidationof violaxanthin to zeaxanthin, showing that the xanthophyllcycle is functioning. Chlorophyll fluorescence analysis of pulvinulesconfirmed that their chloroplasts are capable of both photosyntheticelectron transport and non-photochemical fluorescence quenching,showing that they build up a considerable transthylakoid protongradient in the light. Application of DCMU to excised pulvinulesand laminar discs, as well as to pulvinules of intact, attachedterminal leaflets blocked electron transport and fluorescencequenching. Application of the uncoupler CCCP to intact pulvinulesalso prevented non-photochemical fluorescence quenching. Therate of movement of the low-light-adapted terminal leaflet inresponse to exposure of its pulvinule to overhead red light(500 µmol m^–2 s^–1) was reduced when the pulvinulewas pretreated with DCMU. The pulvinar response to overheadblue light (50 µmol ^–2 s^–1), which is morepronounced than to red light, was not affected by similar pretreatmentwith DCMU. Pretreatment with CCCP caused a short lag in theresponse to red light, but did not affect its subsequent rate.The results suggest that the pulvinar response to red, but notto blue light, requires non-cyclic electron transport and theresulting generation of ATP Key words: Leaf movements, light, non-cyclic electron transport, Phaseolus, pulvinar chloroplasts     

Carbon Dioxide Photoassimilation in Normal-leaved and Mutant Forms of Pisum sativum L.

The reproducible steady-state carbon dioxide (CO₂) photoassimilationpotentials of three mutants and a normal form of pea (Pisumsativum L.) have been compared. The three mutants studied differed markedly in foliar morphology:genotype af af Tl Tl had leaflets converted to tendrils; AfAf tl tl had tendrils converted to leaflets; af af tl tl hadrelatively minute leaflets on a branched petiole. Interest layprimarily in the phenotype with only tendrils since it provideda potential means of reducing the volume of haulm that has tobe rapidly processed in the case of vining peas, and dried inthe case of harvest peas. These mutants had been derived from relatively unimproved cultivars.Before completion of the lengthy backcrossing required to makea full assessment of the value of such mutants an interim studyusing infra red gas analysis indicated that, in terms of CO₂ photoassimilation perunit area of youngest expanded attached leaf of glasshouse-grownplants, the mutants were comparable to normal. The phenotypewith only tendrils was the least efficient of those assayedat utilizing light of an intensity below 100 J m² sec¹ (400–700nm), and on a unit dry-weight basis it was only 18 per centas efficient as a normal-leaved pea. The other mutants werecomparable to normal in this respect. Comparison of CO₂ photoassimilation of glasshouse-grown andfield-grown plants showed them to be similar though they differedin dry weight, transpiration, and dark respiration.     

An efficient method of selecting photoautotrophic cells from cultured heterogeneous cells

A new and efficient method was demonstrated for the establishmentof photoautotrophic cultures of plant cells. Leaf segments ofAtropa belladonna, Datura stramonium and Hyoscyamus niger wereinoculated on sugar-free Linsmaier-Skoog agar medium then aeratedwith 1% CO₂ enriched air under 3,000 to 5,000 lux of illumination.Under these regulated conditions we could select photoautotrophicgreen cells efficiently, and these cells subsequently have grownwell under photoautotrophic conditions. ¹Department of Horticulture and Agriculture, Faculty of Agriculture,Kobe University, Kobe 657, Japan (Received June 12, 1980; )     

An effective time for CO2 gas treatment in overcoming self-incompatibility in Brassica

An investigation was made to determine the effective time forCO₂ treatment in overcoming self-incompatibility in Brassica.CO₂ was effective when supplied to a self-pollinated flowerwhile hundreds of pollen grains were germinating on the stigma.Since the effective time coincides with the attachment of pollentubes to papilla cells, it is thought that CO₂ produces a metabolicchange in these cells during attachement. ¹Part of a thesis submitted for the Dr. of Agr. degree by thesenior author at Tohoku University. ²Present address: Faculty of Agriculture, Kobe University, Nada-ku,Kobe, Japan. (Received December 7, 1972; )     

The Photosynthetic and Respiratory Potential of the Fruit in Relation to Seed Yield of Leafless and 'Semi-leafless Mutants of Pisum sativum L.

The photosynthetic and respiratory net CO₂ exchange potentialof the fruit of Pisum sativum was evaluated in one normal andthree mutant genotypes differing widely with respect to foliagearea and morphology: AfAf.StSt, conventional; AfAf.StSt, vestigialstipules; afaf.St.St, all leaflets replaced by tendrils butstipules normal (here termed ‘semi-leafless’) andafaf.stst, tendrils and vestigial stipules (‘leafless’).Agronomically the latter phenotype offered improvement in resistanceto lodging, crop drying, and harvester through-put. On a dry weight basis, fruits of the leafless mutant were consistentlymore active photosynthetically in terms of CO² uptake from theatmosphere during the initial 18 days post-anthesis than werethe corresponding fruits of the other three phenotypes. Duringthe subsequent 16-day period of seed filling there was no markeddifference between phenotypes and the fruit continuously lostCO² to the atmosphere, but significantly less CO² was lost inthe light (40 k lux) than in the dark. That the fruit of theleafless mutant may therefore benefit from the increase in lightavailable to the fruit within a sward canopy comprised of tendrilsin place of leaflets is discussed. However, there was stilla significant reduction in seed yield associated with the leaflessmutant despite a normal complement of ovule initials. The growthcurves and mean dry weights per seed were not significantlydifferent between phenotypes. In fruit of the conventional phenotypethe attainment of maximal pod wall weight coincided with theinitiation of seed fillingonday 15 after anthesis. In a seriesof isogenic lines the gene af delayed maximal wall developmentby 6 days whilst the gene st markedly lowered the maximum wallweight attainable. Possible causes of yield reduction in theleafless mutant are discussed in relation to the observed actionof these genes in the homozygous double recessive condition.     

Changes in Extracellular pH of the Motor Cells of Mimosa pudica L. during Movement

By using a pH micro-electrode, the changes in the pH of theextracellular fluid in the motor tissue of the main pulvinusof Mimosa pudica were examined. During the rapid movement inducedby dropping ice-cold water on the petiole, the extracellularpH in the lower half shifted from 5.1 to 5.7, and then decreasedgradually to the initial pH within the next 30 min. A similarrise in extracellular pH from 5.1 to 5.4 was also observed inthe upper half. ¹ Present address: Department of Biology, Kobe Women's University,Higashisuma, Kobe 654, Japan. (Received May 18, 1984; Accepted December 6, 1984)     

Effect of EGTA on Nyctinastic Closure Mediated by Phytochrome in Albizzia lophantha

The role of extracellular calcium on nyctinastic closure ofAlbizzia lophantha leaflets has been studied by testing theeffect of ethyleneglycol-bis-(ß-aminoethylether)-N,N,N',N'-tetraaceticacid (EGTA) and its reversibility by calcium. EGTA (1 and 5mM) causes an inhibition of nyctinastic closure and at a concentrationof 1 mM EGTA it decreases the difference between the effectof red light (R) and far-red light (FR) irradiation on leafletclosure. A simultaneous or subsequent supply of CaCl₂ (5 or10 mM) reverses EGTA (5 mM) inhibition on closure as well ascausing an additional promotion of closure. We suggest that external calcium could play a dual role in nyctinasticclosure. Phytochrome control of leaflet closure probably needsexternal Ca²⁺ and, in addition, Ca²⁺ could regulate the closuremechanism by controlling ionic fluxes through the plasma membranein pulvinular motor cells. (Received June 9, 1989; Accepted November 27, 1989)     

The Effect of Osmotic Stress on the Solutes in Guard Cells of Vicia faba L.

We investigated the changes in the levels of solutes in guardcells under osmotic stress. Epidermal strips peeled from Viciafaba L. leaflets were sonicated and incubated in 0.4 M mannitolsolution (osmotic stress) in either light or dark. Stomata wereclosed by osmotic stress. Under osmotic stress, malate accumulatedlight-dependently and sucrose accumulated light-independentlyin the guard cells. The level of K⁺ in guard cells increasedslightly under osmotic stress in the light, although withoutstatistical significance. The levels of all these solutes werereduced by 10 µM ABA treatment. These results suggestthat osmotic stress affects carbon metabolism in guard cells;this metabolic change is different from that caused by ABA alone.Respiratory activity of guard cells decreased under osmoticstress. Therefore, the accumulation of malate and sucrose maybe caused by reduced respiration under osmotic stress. Accumulationof solutes in guard cells by osmotic stress may result in increasedosmotic pressure of guard cells and may play a role in protectionof guard cells from osmotic stress. (Received December 17, 1998; Accepted May 28, 1999)     

Photosynthetic electron transport in Dunaliella tertiolecta (Chlorophyceae) measured by fast repetition rate fluorometry: relation to carbon assimilation

A comparison of photosynthesis-irradiance response curves (P–Eresponse curves) obtained through fast repetition rate (FRR)fluorometry and radiocarbon (¹⁴C) tracer method was made inthe chlorophyte, Dunaliella tertiolecta, grown under differentirradiance conditions. In FRR-based P–E response curveexperiments, actinic light provided by white light-emittingdiodes (LEDs) was increased gradually from 0 to 1500 µmolquanta m^–2 s^–1 and the rate of photosyntheticelectron transport was determined at each light level. Short-termexperiments (20 min) of ¹⁴C-based P–E response curvewere carried out with an improved photosynthetron, which containswhite LEDs as the light source. Irrespective of growth irradiance,the ratios of FRR to ¹⁴C-based initial slopes were almost uniform.The ratios of FRR- to ¹⁴C-based maximum rates were 25–36%higher than those of FRR- to ¹⁴C-based initial slopes. The relationshipbetween electron transport and carbon assimilation was non-linearwith increasing discrepancy towards high actinic light. Thisnon-linear relationship between FRR- and ¹⁴C-based estimatesis primarily due to the effect of physiological processes stimulatedat high levels of light, such as cyclic electron flow and theMehler reaction. The results of this study indicate that theFRR fluorometry can be used as a good indicator of photosyntheticrates from low to middle light levels, but becomes increasinglyquestionable as the maximum photosynthetic rate is approached.The degree to which this relationship is further affected bynutrient-status warrants investigation.     

Donation of Electrons from Cytosolic Components to the Intersystem Chain in the Cyanobacterium Synechococcus sp. PCC 7002 as Determined by the Reduction of P700+

Cells, of Synechococcus sp. PCC 7002 showed a low oxidationlevel of P700 under a far-red light at 6 W m^–2 which inducednearly complete oxidation of P700 in spinach leaves, and a strongerfar-red light was required to observe the oxidation of P700.DCMU did not affect the level of P700⁺² but 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinoneinduced the oxidation of P700 under far-red light, indicatingthat the low oxidation level of P700 was due to the donationof electrons to P700⁺² from the cytosolic respiratory donorsthrough the intersystem chain at the plastoquinone pool. Theelectron transfer from the cytosolic donors to the intersystemchain was inhibited by HgCl₂ but not by antimycin A. The reductionof P700⁺ in Synechococcus cells, after illumination by strongfar-red light was mostly accounted for by the electron flowto the inter system chain from the respiratory donors (t     

Mutant dn Influences Dry Matter Distribution, Assimilate Partitioning and Flowering in Lathyrus odoratus L.

The flowering mutant dn in sweet pea was used as a tool to study¹⁴C-assimilate and dry matter partitioning with respect to nutrientdiversion theories on the control of flower initiation. Wildtype plants (Dn^h) are photoperiodic and exhibit late floweringand profuse basal branching in short days while mutant plants(dn) are day neutral, early flowering and devoid of basal laterals.In short days, dn plants exported a significantly greater proportionof assimilate acropetally than (Dn^h) plants and the upper portionof dn plants had a greater dry weight. These differences werereduced dramatically when basal laterals were excised regularlyfrom the (Dn^h) plants although the difference in flowering remained.However, the effect of dn on resource allocation within theapical region may be more important in regard to flowering thanthe effect on acropetal versus basipetal movement. In shortdays, the dn plants partitioned significantly more resourcesinto their internodes and petioles, and less into their leaflets,than Dn^h plants as shown by dry weight and ¹⁴C-assimilate measurements.These differences were apparent from as early as node 7 up tothe node of flower initiation in dn plants (node 30) and theywere not eliminated by removal of basal laterals from Dn^h plants.Differences between dn and Dn^h plants in partitioning and floweringwere largely eliminated under long days. The fact that in thisspecies a single gene influences both resource allocation andflower initiation lends further support to nutrient diversionhypotheses on the control of flowering. Key words: Assimilate partitioning, branching, flowering, mutant, sweet pea     

A Rapid Method for the Determination of the Leaf Area of Oxalis corniculata L. and Tephrosia purpurea (Linn.) pers.

A rapid simple method has been described for the determinationof leaf area of Oxalis corniculata L. (Oxalidaceae) and Tephrosiapurpurea (Linn.) pers. (Leguminoseae) by evaluating area = a²x0.866and a²x0.335 respectively for the two plants, where a representsa single linear measurement. The relation has been derived afterintegrating the equation of the curve r² = a² cos 2 coveringthe area of each leaflet. Areas of a large number of leaflets have been determined withthe help of two methods (1) Planimeter method, (2) Evaluationmethod. The significance of the difference between the two valuesfor a particular leaf has been statistically examined and itappears that the former method can be replaced by the latterin the case of these plants.     

Biosynthesis, Intracellular Transport and In Vitro Processing of 11S Globulin Precursor Proteins of Developing Castor Bean Endosperm

In vivo labeling experiments to study the biosynthesis of 11Sglobulin in developing castor bean (Ricinus communis) endospermdemonstrated that the subunit polypeptides of the 11S globulinwere synthesized as high molecular weight precursors with heterogeneousmolecular weights. These proglobulin species were not synthesizedconcomitantly during seed maturation. The largest proglobulinwas synthesized from 20 days after anthesis, whereas the smallerproglobulins were synthesized from 30 days after anthesis. Subcellularfractionation of the pulse-labeled endosperm showed that the[³⁵S]methionine label was present in proglobulins in both theendoplasmic reticulum (ER) and dense vesicles shortly afterthe pulse labeling. The label in the proglobulin in ER decreasedduring the chase and appeared in mature globulins associatedwith crystalloids of vacuoles (protein bodies). Proglobulinsin the ER fraction prepared from the pulse-labeled developingendosperm were processed in vitro into globulins by the matrixfraction of protein bodies isolated from the dry castor bean.Overall results indicate that precursor proglobulin moleculessynthesized on rough ER are transported to vacuoles via densevesicles, and are cleaved there by the matrix protease to yieldmature globulin. ¹Department of Botany, University of Maryland, Present address:CollegePark, MD 20742, U.S.A. ²Department of Biology, Faculty of Science, Kobe University,Present address:Rokkoudai, Nada, Kobe 657, Japan (Received June 1, 1987; Accepted December 16, 1987)     

The Role of the Stem in Phloem Loading of Minerals in Lupinus albus L. cv. Ultra

Collections of phloem sap made over a 40-day period from a varietyof locations on nodulated white lupin plants (Lupinus albusL. cv. ultra) showed considerable enrichment with K⁺ and Mg²⁺in the phloem streams destined for the shoot apices or fruitsrelative to the streams arising from the leaflets (up to 5.5times). Sodium showed enrichment in the streams destined forthe roots (up to 2.5 times) but only when present in the watersupply at a high level (3 mM). The stem, in view of its centrallocation in the transport pathway, is seen as an organ capableof redistributing minerals in the phloem independently of photosynthate. Lupinus albus L., lupin, phloem loading, magnesium, potassium, sodium, mineral elements     

Studies on chloroplast development in Chlamydomonas reinhardtii I. Effect of brief illumination on chlorophyll synthesis

When dark grown cells of Chlamydomonas reinhardtii y-1 mutantwere exposed to continuous light, an immediate transformationof small amounts of protochlorophyll(ide), which had been presentin the dark grown cells, to chlorophyll was observed. Afterthis, there was a slow accumulation of chlorophyll lasting for2.5-3 hr before the start of exponential synthesis. Initialaccumulation of chlorophyll was distinctly slower at a highlight intensity (13,000 lux) than it was at moderate intensitiesof light (2,000–5,000 lux). However, the exponential synthesisof chlorophyll started after the same 2.5–3 hr of illumination. A brief pre-illumination of cells followed by incubation indarkness was effective in promoting chlorophyll synthesis undersubsequent continuous illumination at high, as well as moderatelight intensities. Pretreatment alleviated retardation of theinitial chlorophyll accumulation by light of high intensity.The promoting effect of preillumination on chlorophyll synthesiswas sufficient, even when a light impulse as short as 10 secwas given. However, the effect was dependent on length of thedark period after the short pre-illumination. The full extentof this effect was observed when the dark period was about 2.5–3hr long. Further dark incubation gradually decreased the effect. On the basis of these findings, it is assumed that a factor(s)responsible for promotion of chlorophyll (or chloroplast) synthesisin the process of greening of dark grown cells is produced duringthe dark period after a brief pre-illumination, and that thefactor is turned over at a relatively fast rate. The possiblenature of the presumed factor is discussed in relation to chloroplastdevelopment. ¹Present address: Department of Biology, Faculty of Science,Kobe University, Nada-ku, Kobe, Japan. (Received August 18, 1970; )     

Diurnal Rb+ Transport from Roots to the Laminar Pulvinus in Phaseolus vulgaris L.

The primary leaves of kidney bean (Phaseolus vulgaris L.) openunder light and close in the dark by the deformation of thepulvinus resulting from diurnal distribution changes of K⁺,Cl^–, organic acid (or H⁺) and NO₃^–. When Rb⁺ was added as a tracer of K⁺ to the seedlings throughtheir roots, it was transported to the pulvinus cells duringthe light period but not during the dark period. Transpirationoccurred vigorously in the light but almost stopped in the dark.We concluded that Rb⁺ absorbed by the roots was carried to thepulvinus by the transpiration stream. Phaseolus vulgaris L., pulvinus, Rb⁺, diurnal transport transpiration stream     

WATER PERMEABILITY OF THE CELL WALL IN NITELLA

1. A method has been developed to measure the hydraulic conductivityof the wall of the internodal cell of Nitella flexilis.
2. Therate of water penetration through the cell wall varies linearlywith the hydrostatic pressure difference between the two sidesof the wall, showing that water permeability of the cell wallremains independent of the pressure difference applied.
3. Waterpermeability of the cell wall is inversely proportionalto itsthickness It is 30µµmin^–3{dot}atm^–3when the thickness of the wall is 10 µ.
4. Water permeabilityof the cell wall is the same for inward andoutward water flow.The polar water permeability of the entiremembrane system (walland protoplasmic part) of the living celldemonstrated by KAMIYAand TAZAWA (1) is, therefore, due tothe living protoplasmicpart.
5. The ratio of the inward to outward permeability constantsofthe protoplasmic layer alone is higher than that of the entiremembrane system composed of protoplasmic layer and cell wall.

¹ Dedicated to Prof. H. TAMIYA on the occasion of his 60th birthday.The present work was supported in part by a Grant-in-Aid forFundamental Scientific Research from the Ministry of Education. ² Present address: Sh?in Women's College, Kobe. (Received July 21, 1962; )     

Cloning of the Gene Encoding a Protochlorophyllide Reductase: the Physiological Significance of the Co-Existence of Light-Dependent and -Independent Protochlorophyllide Reduction Systems in the Cyanobacterium Plectonema boryanum

Cyanobacteria have two protochlorophyllide (Pchlide) reductasescatalyzing the conversion of Pchlide to chloro-phyllide, a keystep in the biosynthetic pathway of chlorophylls (Chls); a light-dependent(LPOR) and a light-independent (DPOR) reductase. We found anopen reading frame (ORF322) in a 2,131-bp EcoRI fragment fromthe genomic DNA of the cyanobacterium Plectonema boryanum. Becausethe deduced amino acid sequence showed a high similarity tothose of various plant LPORs and the LPOR activity was detectedin the soluble fraction of Esche-richia coli cells over-expressingthe ORF322 protein, ORF322 was defined as the por gene encodingLPOR in P. boryanum. A por-disrupted mutant, YFP12, was isolatedby targeted mutagenesiss to investigate the physiological importanceof LPOR. YFP12 grew as well as wild type under low light conditions(10-25 µE m^–2 S^–1). However, its growth wassignificantly retarded as a result of a significant decreasein its Chl content under higher light conditions (85-130 µEm^–2 s^–1). Furthermore, YFP12 stopped growing andsuffered from photobleaching under the highest light intensity(170 µE m^–2 s^–1). In contrast, a chlL-dis-rupted(DPOR-less) mutant YFC2 grew as well as wild type irrespectiveof light intensity. From these phenotypic characteristics, weconcluded that, although both LPOR and DPOR contribute to Chlsynthesis in the cells growing in the light, the extent of thecontribution by LPOR increases with increasing light intensity;without it, the cells are unable to grow under light intensitiesof more than 130 µ Em^–2s-^–. (Received September 26, 1997; Accepted November 21, 1997)