CHANGES IN THE CONTENT OF GIBBERELLIN-LIKE SUBSTANCES IN RIPENING SEED AND POD OF LUPINUS LUTEUS

The amount of gibberellin-like substances in seed, pod, embryoand "endosperm" of Lupinus luteus in relation to their developmentwas studied by means of the rice seedling test. The amount of gibberellin-like substances per seed increasedremarkably at the first stage of development, when the growthof seed and pod was very slow, and attained its maximum at suchan early stage as the dry weight of seed reached a value ofonly 10 per cent and that of pod 28 per cent of their respectivemaximum levels. Changes in amount of gibberellin-like substancespresent in the pod were very closely related to the changesin growth of pod. Gibberellin-like substances equivalent to0.3 µg gibberellin A₃ per seed and 0.028 µg perpod were found at the 20th day after anthesis, and no activityin mature seed and 0.09 µg per pod at the 50th day. Noactivity was found in the embryo itself at all stages, indicatingthat the gibberellin-like substances in seed occur only in the"endosperm," i.e. in the tissues that will eventually form testain fully ripened stage. (Received December 18, 1962; )     

CHANGES IN THE AMOUNT OF GIBBERELLIN-LIKE SUBSTANCES IN THE SEEDLING OF PHARBITIS NIL WITH SPECIAL REFERENCE TO EXPANSION OF COTYLEDON

Two gibberellin-like substances were separated chromatographicallyfrom the ethanol-extract of germinating cotyledons of Pharbitisnil. In the dark-grown seedling, these substances in the cotyledonincreased in amount gradually with the slow increase in freshweight and area of the tissue, and attained, 4 days after sowing,a maximum level of 0.03 µg gibberellin A₃ equivalent pera couple of cotyledon; in the light-grown seedling, they increasedrapidly with the rapid increase in the fresh weight and area,and attained a maximum content of 0.07 µg gibberellinA₃ equivalent per cotyledon on the 6th day. When the etiolatedseedling was exposed to light, the amount of the gibberellin-likesubstances in the cotyledon increased in parallel with openingof the hypccotyl hook and expansion of the cotyledon. The openingof the hook and increase of the substances occurred in red lightas well as in far-red. Relatively small amounts of the gibberellin-like substanceswere also contained in the germinating hypocotyl and root. The growth of cotyledonary disks was promoted by gibberellinA₃ and the gibberellin-like substances from the expanding cotyledon. The dwarf rice seedling and dwarfs 1, 3 and 5 of maize respondedto the gibberellin-like substances, but dwarf Pharbitis didnot. (Received August 20, 1963; )     

Hormonal and Structural Aspects of Fruit Development in the Orchid, Epidendrum

Endogenous cytokinin and gibberellin-like activity were measuredby bioassay in developing fruit of the orchid Epidendrum ibaguense.Cytokinins decline during the first 30 d after pollination,then begin to accumulate, with very high levels (1–13µ g zeatin eq. g^–1 dry wt. ) in the mature fruitand seed. The major structural change in developing fruit duringthe first 30 d is the ongoing cell division in the fruit wall.By day 30 most ovules have been fertilized and embryo developmentbegins. The increase in cytokinin activity thus coincides withthe onset of embryo development. Gibberellin levels declinein the fruit throughout development, although high activity(0.9 µ g GA₃ eq. g^–1 dry wt. ) is observed in themature seed. The mature embryo shows no obvious structural differentiationinto embryonic axis and cotyledon and no endosperm develops.     

CHANGES IN THE CARBOHYDRATES, PROTEINS AND NUCLEIC ACIDS DURING SEED DEVELOPMENT IN OPIUM POPPY

This investigation is concerned with the major changes in thelevels of carbohydrates, proteins, nucleic acids and some enzymesin the ovules of Papaver somniferum at various stages of seeddevelopment. Of the soluble sugars, fructose and glucose arepresent in large amounts up to the free nuclear stage of theendosperm but decrease rapidly when the latter turns cellular;then sucrose becomes most abundant. At this time the concentrationof insoluble carbohydrates is almost one-seventh that of solubleand the activities of - and ß-amylases are at theirhighest. The nitrogen in the seed is accumulated in two phases,the first coinciding with the development of endosperm and thesecond with the development of embryo. Cell wall formation inthe coenocytic endosperm is accompanied with a marked decreasein the soluble nitrogen. The activity of glutamic-alanine transaminaseincreases concurrently with the increase of protein in the ovules.The maximal RNA content is attained after the ovules have completedtheir increase in size but prior to the deposition of proteinaceousreserves in the endosperm. The amount of DNA increases withthe growth of endosperm but decreases during the maturationof embryo. Both ribonuclease and deoxyribonuclease show twopH optima corresponding to pH 5.5 and 7.5 (ribonuclease) and5.0 and 7.5 (deoxyribonuclease). Their activities seem to becorrelated to the levels of nucleic acids. ¹ Present address: AEC Plant Research Laboratory, Michigan StateUniversity, East Lansing, Mich., U.S.A.     

Identification of Gibberellins in Cotyledonary Embryos of Phaseolus coccineus L.

Gibberellins present in cotyledonary embryos of Phaseolus coccineusL. have been identified by combined gas chromatography-massspectrometry as GA₁, GA₄, GA₅, GA₆; also an unidentified gibberellinwas present. The total amount of gibberellins was estimated by gas chromatographyto be 26.5 µg per g fresh weight; the individual gibberellinscontributed to the total amount as follows: GA₁ 18.4µg,GA₅ 2.6 µg, GA₄ 2.3 µg, GA₆ 1.9 µg, unidentifiedGA 1.3 µg. Data are discussed in relation to previousresults in P. coccineus seed as well as in relation to the embryo-suspensorsystem. (Received December 12, 1985; Accepted July 8, 1985)     

Changes in Levels of Naturally Occurring Gibberellin-like Substances during Germination of Seed of Lycopersicon esculentum Mill.

Naturally occurring gibberellin-like substances possessing acidic,basic, and neutral properties were detected, by paper partitionchromatography, in ethanolic extracts of tomato seed and ofetiolated seedlings after 72 and 116 hours' growth. Dwarf maizemutants of the d-1 and d-5 types, ‘Meteor’ pea seedlingsand young ‘Potentate’ tomato plants were used asbioassay material. Hydrolysis of seed and seedling proteinsby ficin in phosphate buffer, pH 6.2, after removal of ethanol-solublesubstances, liberated more and different ‘bound’gibberellin-like substances. It is suggested that protein hydrolysisduring germination is an important means of liberating thesesubstances at different stages of seedling development. Acidic substances were present in all the extracts prepared,but in general two with Rfs 0.25 and 0.55 in iso-propanol: ammonia:water : : 10:1:1 v/v were differentiated on d-2 and d-5 maizerespectively. Neutral substances in dry seed extracts chromatographedin the same solvent, had Rfs of 0.05, 0.35, and 0.95 and thesewere found only in the ethanolic (‘free’) extracts.They were active on d-1 and d-2 maize and ‘Meteor’pea. Basic gibberellin-like substances with Rfs of 0.05 and0.35 were found in ‘free’ extracts of both dry seedand etiolated seedlings after 116 hours' growth which were activeon d-2 maize only. Two others with Rfs 0.45 and 0.95 were extractedfrom seedlings after 72 hours' growth and these were activeon young ‘Potentate’ tomato plants. It is suggested that certain gibberellin-like substances, capableof reversing dwarfism in test plants, may be responsible formorphogenetic or other responses not involving stem extensionin the parent species. Changes were found in the levels of gibberellin-likesubstances but there was no evidence of changes in levels ofseed inhibitors relative to seed growth substances.     

CHANGES IN EMISSION SPECTRA OF PHOTOSYNTHETIC PIGMENTS IN VIVO

1. The effects of 3-(4'-chlorophenyl)-1, 1-dimethylurea (CMU)onthe fluorescence of photosynthetic pigments in vivo wereinvestigatedin blue-green, red and brown algae and in isolatedspinach chloroplasts.CMU caused an increase in steady statelevel of fluorescenceof chlorophyll a, but did not influencethe fluorescence ofphycobilins. The spectrum of the fluorescenceincrement hada peak at 685 m/µ and a shoulder at 730–740mµ.These two bands probably arise from chlorophyll a(C_f684) belongingto pigment system II.
2. On excitation of chlorophylla in a red alga, Porphyra yezoensis,a fluorescence band witha peak at 720 mµ was observedbesides a shoulder at 685mµ. The 720 m band is inferredto arise from chlorophylla (probably, C_f-1) in pigment systemI.
3. On addition of CMUto the algal cells, the induction of fluorescencewas modifiedto take a simple time course. The induction wasobserved onlywith respect to the fluorescence of chlorophylla, but not inthe fluorescence of phycobilins. The spectrumof the "transient"fluorescence showed two emission bands ofchlorophyll a at 685mµ and 740 mµ, and was quitesimilar in form tothe spectrum of the CMU-caused increase insteady state fluorescence.
4. These facts were interpreted in terms of the correlation offluorescence of chlorophyll a and the photochemical reactionsof photosynthesis

(Received July 20, 1967; )     

Phase Change in Hedera helix L: I. GIBBERELLIN-LIKE SUBSTANCES IN THE TWO GROWTH PHASES

Gibberellin-like substances were extracted from all parts ofthe juvenile and adult growth phases of Hedera helix. A numberof extraction techniques were employed and estimatos of theamount of gibberellin-like activity present were made usingthree bioassays. Gibberellin like substances were present inapproximately equal amounts in leaves at the two growth phasesand appeared to be localized, in the main, in chloroplasts.Apical ‘buds’ of the juvenile phase were shown tocontain higher levels of gibberellin-like substances than theiradult counterparts. Roots, which under normal circumstancesare associated only with the juvenile phase, contained highlevels of extractable gibberellin-like substances.     

An Analysis of Seed Development in Pisum sativum III. The Relationship Between the r Locus, the Water Content and the Osmotic Potential of Seed Tissues in vivo and in vitro

The water content and osmotic potential (₂) of the differentparts of the pea fruit have been measured during developmentof the seed in two lines near-isogenic except for the r locus.During the early development of both genotypes, the testa possesseda more negative ₂ than either embryo, endosperm or pod while,at stages when liquid endosperm was present, the embryo alwaysmaintained ₂, more negative than the endosperm. A clear effectof the r locus on water content and ₂ was only observed in embryotissue — wrinkled (rr) embryos possessing more water andmaintaining a more negative ₂ than round (RR) for most of thedevelopmental period studied. The more negative ₂ of wrinkledembryos correlated with their greater uptake of water in vivo. When cultured in vitro, the embryos of both genotypes showedmaximum growth (fresh or dry weight) if 10 per cent sucrosewas added to the medium (equivalent to about — 1.2 MPa).Round embryos, however, increased in weight more than wrinkledat all sucrose concentrations examined. Cultured embryos maintaineda similar or more negative ₂ than their surrounding medium;wrinkled more negative than round. Embryo culture, osmotic potential, Pisum sativum, pea, r locus, seed development, tissue culture, water content     

EFFECTS OF VARIOUS FACTORS ON THE INCREASE OF {alpha}-AMYLASE ACTIVITY IN RICE ENDOSPERM INDUCED BY GIBBERELLIN A3

The increase of -amylase activity in embryoless rice endosperminduced by the addition of gibberellin A₃ was examined undervarious conditions with an aim to establish a bioassay methodfor gibberellins. Sterilized embryoless rice endosperms were incubated in a testtube containing 0.2–1.0 ml of test solution for 4 daysat 30. -Amylase activity in the endosperm was determined bymeasuring digestion of added starch. The increase of amylaseactivity during the incubation was not affected by the additionof various vitamins, amino acids, organic acids, protease, sucrose,indoleacetic acid or kinetin. Helminthosporol, helminthosporicacid and sclerin (1–10 µg/ml) had weak promotingeffects. Under appropriate conditions, 10^–5 µg/mlof gibberellin A₃ could be detected. In double logarithmic plot,the increase in the enzyme activity was proportional to thegibberellin A₃ concentration in the range from 10^–5 to10^–2 µg/ml. (Received April 13, 1966; )     

VINYL PHEOPORPHYRIN A5 IN THE INNER SEED COATS OF PUMPKINS

The pigments contained in the inner seed coats of Cucurbitapepo, C. maxima and C. moschata were investigated spectroscopicallyusing both intact tissues and their ether extracts. The pigment(s) contained in C. pepo and C. maxima was inferred to be "protochlorophyll"and/or Mg vinyl pheoporphyrin a₅ which are indistinguishablefrom each other by mere spectroscopic observations. The coatsof C. moschata were found to contain Mg-free vinyl pheoporphyrina₅ which has so far been reported to exist only in some Chlorellamutants. The spectra of these pigments in intact tissues wereremarkably different from those of the ether extracts, indicatingthe profound difference in their physical states in vitro andin vivo. It was discussed that the "protochlorophyll b" reportedearlier by SEYBOLD and EGLE might have been vinyl pheoporphyrina₁. (Received July 14, 1960; )     

Gibberellin-like Activity in the Developing Avocado Fruit

Gibberellin-like activity in avocado (Persea americana) fruit extracts was measured by the barley endosperm bioassay. Fruit tissues were anlyzed separately during fruit growth. The level of the activity found was very high in the endosperm and in the seed coats, but decreased in the latter during fruit growth. No measurable gibberellin-like activity was detected in the mesocarp or in the embryo. It is assumed that the seed coats are a site of production of gibberellin-like substances in the avocado fruit.     

The Movement of Ions to the Xylem Exudate of Maize Roots: II. A COMPARISON OF THE ELECTRICAL POTENTIAL AND ELECTROCHEMICAL POTENTIALS OF IONS IN THE EXUDATE AND IN THE ROOT CELLS

The electrical potential differences (E_ex) and the electrochemicalpotential differences for potassium and chloride in the xylemexudate of excised maize roots were determined and the valuesobtained indicated active transport of both ions to the xylemexudate. The potential differences were depolarized by increasesin the external KCl concentration ([KCl]_o), and E_ex was a linearfuction of log[KCl]_o. The corresponding data for the cell vacuolesof the same roots were also determined and trends similar tothose found for the exudate were obtained. The electrochemicalpotential differences between the cell vacuoles and the externalsolution for potassium (µ_rac.k) and chloride (µ_vac.Cl)indicated that both ions were actively transported to the vacuoles.Both µ_vac.k and µ_vac.cl decreased in a linear manneras [KCl]_o was increased. However, in spite of the similar trendsin both sets of data, the values of the electrical potentialand chloride electrochemical potential differences were alwaysmarkedly larger for the vacuoles than for the exudate. For potassium,there were no significant differences. It is concluded thatthe discrepancy between the data for the vacuoles and the wholeroots does not support the idea that the exuding root systembehaves like a single cell, as has been suggested by some workers.     

Paradoxical helium and sulfur hexafluoride single-breath washouts in short-term vs. sustained microgravity

Lauzon, Anne-Marie, G. Kim Prisk, Ann R. Elliott, SylviaVerbanck, Manuel Paiva, and John B. West. Paradoxical helium andsulfur hexafluoride single-breath washouts in short-term vs. sustainedmicrogravity. J. Appl. Physiol. 82(3):859-865, 1997.During single-breath washouts in normal gravity (1 G), the phase III slope of sulfur hexafluoride(SF₆) is steeper than that ofhelium (He). Two mechanisms can account for this:1) the higher diffusivity of Heenhances its homogeneous distribution; and2) the lower diffusivity ofSF₆ results in a more peripherallocation of the diffusion front, where airway asymmetry is larger.These mechanisms were thought to be gravity independent. However, weshowed during the Spacelab Life Sciences-2 spaceflight that insustained microgravity (µG) theSF₆-to-He slope difference isabolished. We repeated the protocol during short periods (27 s) of µG(parabolic flights). The subjects performed a vital-capacityinspiration and expiration of a gas containing 5% He-1.25%SF₆-balanceO₂. As in sustained µG, thephase III slopes of He and SF₆decreased. However, during short-term µG, theSF₆-to-He slope differenceincreased from 0.17 ± 0.03%/l in 1 G to 0.29 ± 0.06%/l inµG, respectively. This is contrary to sustained µG, in which theSF₆-to-He slope difference decreased from 0.25 ± 0.03%/l in 1 G to 0.01 ± 0.06%/lin µG. The increase in phase III slope difference in short-term µGwas caused by a larger decrease of He phase III slope compared with that in sustained µG. This suggests that changes in peripheral gasmixing seen in sustained µG are mainly due to alterations in thediffusive-convective inhomogeneity of He that require >27 s of µGto occur. Changes in pulmonary blood volume distribution or cardiogenicmixing may explain the differences between the results found inshort-term and sustained µG.

     

STUDIES ON THE CONDITIONS FOR GIBBERELLIN ASSAY USING RICE SEEDLING

Of many varieties of rice plant tested, a dwarf mutant "Tamanishiki"was the most suitable material for gibberellin assay. It respondsto neither kinetin nor indoleacetic acid in the concentrationrange from 0.01 to 10 µg/l. Seeds were sterilized withsodium hypochiorite solution (chlorine content, one per cent)for one hour and soaked in water for 2 days. Germinating seedsselected for uniformity were then placed in test tubes containing1–3 ml of test solution, and the length of the 2nd leafsheath of the seedlings was measured after 7 days of incubation.The best conditions for the incubation were: pH of the testsolution, 4; temperature, 30°; continuous illumination ofintensity stronger than 50 lux. Under the best conditions statistically significant growth promotionwas found with 7 seedlings grown in one ml of the test solutionwhich contained 0.001 µg gibberellin A_s In double logarithmicplot, the growth promotion over control increased linearly withincreasing gibberellin concentration up to 1 µg in oneml test solution. (Received March 7, 1963; )     

On the causes of interspecific differences in the growth-irradiance relationship for phytoplankton. II. A general review

The causes of interspecific differences in the µ-l relationshipare examined in the context of a mechanistic model which relatesµ to irradiance in terms of six factors:, k_c photosyntheticquotient (PQ), Chl a:C, respiration and excretion. The effectof cell size on the light saturated growth rate is also considered.It is shown that photosynthetic efficiency and PQ exhibit remarkablylittle interspecific variability, and average 0.024 ±0.005 µg C(µg Chl a)^–1 h^–1 (µEm^–2 s^–1)^–1 and 1.5 ± 0.2 mol 02 molC^–1 (when NO₃^– is the nitrogen source) respectively.Two useful relationships were derived: (i) between growth efficiency,_g and Chl a:C at µ. = 0; (ii) between the compensationintensity, I_c and the Chl a-specific maintenance respirationrate. Both relationships were independent of temperature anddaylength. Species best adapted to growth at low light werefound to exhibit high Chl a:C ratios and low maintenance respirationrates. As a group, diatoms were consistently the best adaptedfor growth at low irradiance. Chiorophytes, haptophytes, chrysophytesand cryptophytes were intermediate in their performance at lowirradiance. Dinoflagellates exhibited extreme diversity, withspecies spanning the spectrum from very good performance atlow irradiance to very poor. A new µ_max-cell carbon relationshipis given based on growth rates normalized to 15°C. Evidenceis presented to show that noise in this relationship can besignificantly reduced by using only carbon-specific growth ratesand using only data for species grown at the same daylength.     

Effects Produced on Tomato Plants, Lycopersicum esculentum, by Seed or Root Treatment with Gibberellic Acid and Indol-3yl-Acetic Acid

Growth in lengths of tomato stems and leaves was acceleratedby 5.0 µg gibberellic acid (GA₂) applied to the seed,or by 5.0, 0.5, and 0.05 µg given to the roots. Treatmentwith 5.0 µg also decreased bud number and lengthened thetime between bud appearance and fruit formation on the firsttruss by 1–8 days. Smaller amounts applied to roots shortenedthis time by 1–4 days. Indol-3yl-acetic acid at 0.5 µghad no effect, nor was simultaneous application of GA₃ and IAAto the roots more effective than GA₃, alone. Single applicationsof very small amounts of GA₃ to seeds or seedling roots thusproved capable of changing growth-rates of stems, leaves, andtrusses.The effects of treating tomatoes with GA₂ and with culturesof Azotobacter chroococcum, which contain small amounts of GA₃,and IAA, are compared.     

UPTAKE OF ACETATE BY NEOTRICULA APERTA (GASTROPODA: POMATIOPSIDAE), THE SNAIL HOST OF SCHISTOSOMA MEKONGI IN THE LOWER MEKONG BASIN

All three races of Neotricula aperta, an epilithic, schistosometransmitting, snail of the Mekong and Mul rivers of NortheastThailand and southern Laos, were found to take up acetate froma dilute solution. After 48 h incubation the mean specific netuptake rates (µmol^–1 g^–1 h^–1), from750 µM acetate, were: 1.86, -race; 1.39, ß-raceand 3.25, y-race. Over 48 h the snails were able to achievereductions in the ambient acetate concentration of up to 60%.Incubations under bacteriostatis suggested that bacteria arenot involved in the uptake of acetate by N. aperta. The uptakecharacteristics conform to the Michaelis-Menten model. The transportconstants, J_max (µmol^–1 g^–1 h^–1) andK_t (µM) were 1.10 and 102 respectively (-race). Racialdifferences in uptake characteristics are discussed in relationto micro-habitat differences. HPLC indicated concentrations of acetate in y-N. aperta microhabitatsof around 325 µM. This suggests a pool size sufficientto satisfy only 6% of the snail's basal metabolic rate (BMR).Levels within the epilithic aufwuchs, however, are probablyhigh enough to provide for more than 50% of the BMR. The possible role of acetate in the energy metabolism of N.aperta is discussed. Short-chain carboxylic acids (such as acetate),arising from the decomposition of the aufwuchs, could representsources of fermentable organics that may be taken up by N. apertasnails and used to supplement their nutrition during times offood shortage. However, further investigations involving ¹⁴C-labellingtechniques are required. The findings of this investigationhave implications for the chemical ecology and life-cycle ofN. aperta. (Received 16 June 1994; accepted 28 July 1994)     

THE RATIO OF CALCAREOUS AND ORGANIC SHELL COMPONENTS OF FRESHWATER SPHAERIID CLAMS IN RELATION TO WATER HARDNESS AND TROPHIC CONDITIONS

Shells from 14 populations of sphaeriid clams including Sphaeriumstriatinum, S. simile, Pisidium walkeri, Musculim partumeiumand M. iransversum were analyzed for organic carbon (µgCmg^–1 shell), nitrogen (µg,N mg^–1 shell) andCaCO_s (%CaCO₃ of total clam dry weight). Habitat waters wereassessed for total hardness (expressed as ppm CaCo₃), ppm Ca,ppm Mg, conductivity (µmho) and suspended organic Carbon(µgCl^–1). For all populations, shell organic C andN are positively correlated and there is an inverse relationshipbetween the amounts of shell CaCO₃ and shell organic carbon.Trophic considerations give the best correlation with shelltype at the generic level of consideration since species ofMusculium are found at the opposite end of the trophic scale(eutrophic) from all other populations studied. For S. striatinum,the most extensively studied species, the amount of shell CaCO₃is inversely related to water hardness. The selection of shellsin the Sphaeriidae is discussed in relation to structural-functionalneeds and habitat conditions ¹ Present Address: Department of Biology, Syracuse University,Syracuse, New York 13210, U.S.A. ² Present Address: Department of Zoology, Miami University,Oxford, Ohio 45056, U.S.A. (Received 5 December 1978;     

COMPONENTS OF THE ELECTRON-TRANSFERRING SYSTEM IN ACETOBACTER SUBOXYDANS AND RECONSTRUCTION OF THE LACTATE OXIDATION SYSTEM

1. Cytochromes a₁⁵⁹⁰, b⁵⁶⁰, c₁⁵⁵⁴ and c₁⁵⁵² were isolated andpurifiedfrom a strain of Acetobacter suboxydans. The proceduresusedwere described in detail.
2. The main cytochrome band at550-560 mµ in intact cellssplitted at liquid air temperatureinto two bands, 551 mµ(strong) and 559 mµ (weak).
3. Optical and physiological properties of the four cytochromeswere investigated. Lactic dehydrogenase activity was found tobe associated with cytochrome c₁⁵⁵⁴. The two c₁-type cytochromes,especially cytochrome c₁⁵⁵⁴, persisted in their reduced formafter the purification through many steps.
4. By some combinationsof isolated components reconstruction ofthe oxygen uptake systemcould be realized.
5. The oxygen-consuming activity of purifiedoxidase preparationswas accelerated by a-tocopherol but notby Emasoll 4130 andTween 80.
6. Some discussions were made onthe nature of terminal oxidase,the role of cytochrome c₁⁵⁵²in the electron-transport system,and persistence of reducedstate of c₁-type cytochromes.
7. A possible scheme of the electron-transferringsystem of Acetobactersuboxydans was presented.

(Received May 16, 1960; )