Glycosylation sites identified by solid-phase Edman degradation: O-linked glycosylation motifs on human glycophorin A

The human red blood cell sialoglycoprotein, glycophorin A (GpA),contains a ‘mucin-like’ extensively O-glycosylatedextracellular domain which carries the MN blood group antigens.We have revised the sites of O-glyccsylation in the extracellulardomain of GpA by automated solid-phase Edman degradation, whichallowed positive identification and quantitation of O-glycosylatedSer and Thr residues, as well as the single N-glycosylationsite. One N-linked and 16 O-linked sites were identified. Carbohydratewas absent on Ser 1, Ser14, Ser15, Ser23, Thr28 and Thr58 inGpA. We propose that the glycosyltransferases present in erythrocytesrecognize specific flanking sequences around potential O-glycosylationsites. All 16 O-glycosylation sites are explained on the basisof four motifs. Three motifs are associated with Thr-glycosylation:Xaa—Pro—Xaa—Xaa where at least one Xaa = Thr;Thr—Xaa—Xaa—Xaa where at least one Xaa = Thr;Xaa—Xaa—Thr—Xaa where at least one X = Argor Lys. The fourth motif is associated with Ser-glycosylation:Ser—Xaa—Xaa—Xaa where at least one Xaa = Ser.These simple rules explain the glycosylation (or lack of it)on 21 of 22 Ser/Thr in the extracellular domain of GpA. glycophorin A O-glycosylation motif solid-phase Edman degradation     

Physical Analysis of Tissue Mechanics in Amphibian Gastrulation

During morphogenesis, intercellular attachments constrain cellmobility so that embryonic tissues may (i) deform as solid sheetsof tightly bound cells, (ii) disperse to migrate as separatecells or (iii) flow as multicellular liquids (in which cellsremain aggregated yet can still slide past one another). Bymodelling deep germ layers as multicellular liquids in amphibiangastrulation, Davis and I have predicted, and then confirmedexperimentally, (i) the area-invariance of deep-germ-layer surfacetensions in vitro, (ii) spontaneous cell slippage in deformeddeep—germ—layer cell aggregates and (iii) correlationsof tissue surface tensions with tissue positioning in deep-germ-layercell-sorting and aggregate—fusion experiments. Liquid—tissueflow involves intercalations of interior cells into expandingtissue interfaces (or withdrawal of surface cells from shrinkingtissue interfaces). Tissue surface tensions are macroscopicreflections of the microscopic, tissue—specific adhesivedifferentials which direct these cell translocations. Such adhesivedifferentials may act independently of, or together with, activecell—shape changes, chemotaxis, contact guidance and/orhaptotaxis in controlling embryonic tissue rearrangements. Deepcell intercalations in vivo occur throughout amphibian gastrulation:during ectodermal epiboly; during marginal—zone extensionand convergence (and therefore blastopore closure); during mesodermalinvolution; and probably during the anterior spreading and axialextension (and therefore dorsal convergence) of involuted mesoderm.Tissue—surface—tension measurements may help determine(i) which of these cell—intercalation processes are activeand which are passive, (ii) the specific contributions of variousmicroscopic cell properties to the regulation of liquid—likegerm—layer assembly and (iii) similarities and differencesbetween in vitro and in vivo control mechanisms governing amphibiangastrulation.     

Sexuality in Diploid and Hexaploid Races of Mercurialis annua L

The variation in sex expression found between individuals ofMercurialis annua agg. is described, distinction being madebetween ‘regular’ and ‘irregular’ monoecism.There is shown to be a relationship between sexuality, morphology,and chromosome number. M. annua sensu stricta and M. annua var. ambigua are shown tobe cytologically distinct, the former being diploid, the latterhexaploid. It is recommended that M. annua var. ambigua be regardedas a species—M. ambigua—as originally proposed byLinnaeus fils. Past investigations into the inheritance of sex are reviewedand results of further investigations into the inheritance ofmonoecism in M. annua var. ambigua are presented. Evidence obtainedsupports the hypothesis that the allosome constitution of maleplants is XXXXXY and that of monoecious plants is XXXXXX.     

Beware of mis-assembled genomes

With hundreds of genomes now in GenBank, researchers might beforgiven for assuming that genome sequence data are correct,at least at a large scale. Certainly there might be errors atsome small rate, perhaps 1 in 50 000 or 100 000 bases (Schmutz et al., 2004;Read et al., 2002), but at a large scale thesegenomes are put together correctly, are not they? Well, notalways. We have been looking at the assemblies of large genomes forseveral years now, and for every ‘draft’ genomewe look at, we find hundreds—and sometimes thousands—ofmis-assemblies. These include regions where a genome is incorrectlyre-arranged as well as places where large chunks of DNA sequenceare simply     

Freezing avoidance mechanism of primordial shoots of conifer buds

Excised winter buds of very hardy fir supercooled to —30or — 35?C, though primordial shoots excised from thesewinter buds (freezing point: about —5.5?C) supercooledonly to —12 to — 14?C. Also, excised primordialshoots did not tolerate freezing, but were rather resistantto desiccation. Differential thermal analysis (DTA) of primordialshoots revealed that the capability of supercooling increasedwith decreasing water content and that no exotherm could bedetected in the primordial shoots with a water content belowabout 20%. When excised whole buds were cooled very slowly,the exotherm temperature shifted markedly to a lower value andthe exotherm became much smaller. Also, masses of needle icewere observed, mainly beneath the crown of the primordial shoot.From these results, it may be concluded that most of the waterin primordial shoots gradually migrates out through the crownand freezes as the temperature decreases (extraorgan freezing),which enables primordial shoots to survive at very low temperatures.Winter buds of Abies balsamea held at — 20?C for 30 daysand then slowly cooled down to —50 or —60?C remainedalive. Thus, there seems to be no low temperature limit to thisfrost avoidance mechanism, if the primordial shoots can resistintensive freeze-dehydration. Low temperature exotherms wereobserved in all genera which belong to Abietoideae and Laricoideaeof Pinaceae, all of which have a crown in the primordial shoots,but not in other conifers. ¹ Contribution No. 2037 from the Institute of Low TemperatureScience. (Received June 25, 1979; )     

Grazing of echinoderm larvae (Paracentrotus lividus and Arbacia lixula) on naturally occurring particulate matter

Larvae (72 hr old) of P. lividus and A. lixula grazed on varioussuspensions of natural particulate matter with a size rangeof 2 to 30 microns, and on two species of algae (Phaeodactylumtricor-nutum and Nitzschia sp.) — Larvae graze most in the size range where the particleconcentration is highest. — If larvae deplete certain size categories of particlesthey then graze other size ranges in which the concentrationis still high. — The grazing rate of the two species varied between 988and 91.949 µm³ per pluteus per hour. — For A. lixula larvae the grazing rate increases withincreasing temperature to a maximum at 22°C.     

Metabolism of (--)-kaurene-3H and (--)-kaurenol-3H during the photoperiodic floral induction in Pharbitis nil

(—)-Kaurene-U-³H was metabolized in seedlings of Pharbitisnil, a short-day plant, to labeled ( — )-kaurenol, ( —)-kaurenal, ( — )-kaurenoic acid, and unidentified polarsubstances, in this sequence. No significant effect of photoperiodicfloral induction upon the metabolism of ( — )-kaurene-U-³Hor ( — )-kaurenol-U-³H was observed, which suggests that( — )-kaurene metabolism is not involved in photoperiodicfloral induction ¹This work was supported in part by grants from the Ministryof Education. (Received September 24, 1970; )     

Resting membrane potential and action potential of Nitella expansa protoplasts

Inside negative membrane potentials were observed for protoplastsobtained from Nitella expansa leaf internodal cells in mediacontaining 1 to 100 mM CaCl₂ using the microelectrode technique.The potential values were less negative than the membrane potentialof intact N. expansa leaf internodal cells. In addition, anaction potential consisting of two components—a fast componentand a slow component—was induced by electrical stimulationfor the protoplasts as well as the intact cells. (Received December 18, 1979; )     

Analysis of the Promoter of the Auxin-Inducible Gene, parC, of Tobacco

The auxin-responsive region (AuxRR) in the promoter of the parCgene was analyzed in transgenic tobacco plants in which the5' flanking region of the parC promoter was placed upstreamof the gene for rß-glucuronidase (GUS). The AuxRRwas located between nucleotides (nt) –226 and –54.Detailed dissection of this segment revealed that the presenceof the non-contiguous sequences from nt –226 to –151and from nt –84 to –54 was required for the expressionof the auxin responsiveness of the parC promoter. The sequencefrom nt –226 to –151 was found to contain a sequencewhich resembles the as-1 element in the 35S promoter of cauliflowermosaic virus (CaMV). Although it has been reported that theas-1 element is involved in auxin responsiveness [Liu and Lam(1994) J. Biol. Chem. 269: 668], we showed that introductionof a point mutation into the as-1-like sequence completely eliminatedauxin responsiveness, a result that suggests that the sequenceis indispensable for auxin responsiveness. However, the presenceof the as-1-like sequence alone was not sufficient for auxinresponsiveness, since the segment (nt –226 to –84)that included the as-1-like sequence failed to confer auxinresponsiveness on the core promoter. It is possible that thetwo separately located sequences play specific roles in interactionswith trans-factors that are required for the expression of theauxin responsiveness of the parC promoter. (Received March 11, 1996; Accepted July 9, 1996)     

Mineral nutrition, of cultured chlorophyllous cells of tobacco VII. Effects of the NH4/NO3 ratio and of Cl in the media on the growth and ionic balance of cells

NG, a strain of cultured tobacco cells of Nicotiana glutinosahad high growth rates and carboxylate contents (C—A) of100 to 130 meq/100 g of dry cells on media containing 42 meqNO₃/liter as the sole N source. (C—A) is the amount ofinorganic cations minus inorganic anions in meq per 100 g ofdry cells. NG, cultured on media containing NH₄ 10+NO₃ 42 in meq/liter,had lower growth rates and lower (C—A) values as comparedwith NG on media containing NO₃ as the sole N source. NG, cultured on media containing NH₄ 30+NO₃ 42 in meq/liter,had high growth rates and (A—C) values of 22 to 53 meq/100gof dry cells. In this case, the (A—C) content may correspondto organic cations, basic organic N compounds such as free asprotein-bound basic amino acids. The easily absorbed Cl mayhave been required maintain good growth conditions such as ionicbalance and a favorable pH in the cells. Thus cultured cells of Nicotiana glutinosa may have physiologicaladaptability against variations in a relatively wide range of|C—A| contents [|C—A| being the absolute valuesof (C—A)]. (Received May 15, 1980; )     

Two Types of Embryo Sac Development in Scrophularia himalensis Royle

The development of the embryo sac in Scrophularia himalensisRoyle (tribe Cheloneae, Scrophulariaceae) has been investigated.The ovules are anatropous, unitegminal and tenuinucellar. Twotypes of embryo sac development—the monosporic (polygonumtype) and the bisporic (allium type)—coexist in this species.This is the first record of its kind in Scrophularia as previousworkers reported only the Polygonum type of development in thetribe Cheloneae.     

Variation in the Durations of the Photoperiod-sensitive and Photoperiod-insensitive Phases of Development to Flowering Among Eight Maturity Isolines of Soyabean [Glycine max (L.) Merrill]

In soyabean [Glycine max (L.) Merrill] the period between sowingand flowering is comprised of three successive developmentalphases—pre-inductive, inductive and post-inductive—inwhich the rate of development is affected, respectively, bytemperature only, by photoperiod and temperature, and then againby temperature only. A reciprocal-transfer experiment (carriedout at a mean temperature of 25°C) in which cohorts of plantswere transferred successively between short and long photoperiodsand vice-versa showed that eight combinations of three pairsof maturity alleles (E₁/e₁, E₂ /e₂, E₃ /e₃) had their greatesteffect on the duration of the inductive phase in long days.This phase was increased with the increasing photoperiod sensitivityinduced by the different gene combinations, and ranged fromabout 27 to 54 d according to genotype. In a short day regime(11·5 h d^-1), less than the critical photoperiod, theduration of the inductive phase was brief—requiring about11 photoperiodic cycles in the less photoperiod-sensitive genotypesand only about seven cycles in the more sensitive ones. Thematurity genes also affected the duration of the two photoperiod-insensitivephases; these durations were positively correlated with thephotoperiod-sensitivity potential of the gene combinations.The largest effect was on the pre-inductive phase which variedfrom 3 to 11 d, while the post-inductive phase varied from about13 to 18 d. As a consequence of these non-photoperiodic effectsof the maturity genes, even in the most inductive regimes (daylengthsless than the critical photoperiod) the time taken to flowerby the less photoperiod-sensitive combinations of maturity geneswas somewhat less than in the more sensitive combinations—rangingfrom about 28 to 34 d. The genetic and practical implicationsof these findings are discussed.Copyright 1994, 1999 AcademicPress Glycine max (L.) Merrill, soyabean, maturity genes, isolines, flowering, photoperiod     

EFFECT OF DIFFERENT MOLLUSC ASSOCIATIONS ON TARGET MOLLUSC GROWTH AND PARASITE CERCARIAL PRODUCTION IN THE TRIPLE SYSTEM: BULINUS WRIGHTI-SCHISTOSOMA BOVIS AND MELANOPSIS PRAEMORSA

The influence of different mollusc associations in the triplesystem: Bulinus wrighti (target mollusc: TM)—Schistosomabovis (parasite)—Melanopsis praemorsa (non-target mollusc:NTM) is analysed. The results show that: (i) for a TM/NTM densityratio of 1/1, the NTM stimulates both growth of the TM and parasitecercarial production; (ii) for a TM/NTM density ratio of 1/5,the effects of the NTM on TM growth and cercarial productionwere not additive and were less marked than for the 1/1 ratio.Hypotheses on a chemical explanation for these density-dependenteffects are discussed. (Received 30 December 1991; accepted 9 March 1992)     

Physiological and Ecological Studies in the Analysis of Plant Environment: VII. An Analysis of the Differential Effects of Light Intensity on the Net Assimilation Rate, Leaf-Area Ratio, and Relative Growth Rate of Different Species

Since relative growth rate is the product of net assimilationrate and leaf-area ratio (leaf area/plant weight), it followsthat if the effects of shading on both net assimilation rateand leaf-area ratio can be expressed mathematically, then therelationship between light intensity and relative growth ratecan be derived from the product of the two mathematical expressions. For all the ten species investigated in field and pot cultureexperiments, it has been found that during the early vegetativephase both the changes in leaf-area ratio and net assimilationrate, over the range of 0·1 to full daylight, are linearlyrelated to the logarithm of the light intensity. In consequence,the relationship between relative growth rate and the logarithmof light intensity—being the product of the two linearregressions—is curvilinear. For species of shady habitats (Geum urbanum, Solamun dulcamara)neither the levels of assimilation rate nor the ‘compensation-point’values are very different from those of the eight species fromopen situations (e.g. Hordeum vulgare, Pisum sativum, Fagopyrumesculentum). Nevertheless the intensity at which growth rateis maximal varies between species: it is 0•5 for G. urbanum,0•7 for H. annuus, full daylight for F. esculentum, whilefor Trifolium subterraneum the calculated value is 1·8daylight. Such specific differences can be largely accountedfor in terms of the differences in leaf-area ratio at the differentlight levels. On the basis of this analysis of the light factor, a ‘shade’plant is best redefined as a species in which a reduction ofthe light intensity causes a rapid rise in the leaf-area ratiofrom an initial low value in full daylight: for a ‘sun’plant the converse definition holds.     

Reproduction rates and secondary production of three species of the rotifer genus Synchaeta in the estuarine Potomac River

Rotifers are a relatively well-studied component of lacustrinesystems but their role is only poorly understood in estuaries.Three species of the genus Synchaeta—S. baltica, S. triophthalmaand S.cecilia—dominate the cold-water assemblage of rotifersin Chesapeake Bay. Laboratory experiments were conducted todetermine the temperature dependence of egg development time(EDT) for each species; EDT varied over an approximate rangeof 90–9 h as temperature (T) varied from 2 to 22°C.The EDT/temperature relationships could be closely fitted bya simple polynomial equation of the form log(EDT) = a + b(logT) + c(log T)² for each species. Natural populations of thesethree rotifers were sampled during a cruise in the Potomac River(7–11 March 1983). Estimates of specific reproductiverates (b) were calculated based on the previously defined EDT/temperaturerelationship and the observed ratio of eggs/rotifers for eachspecies. The two most abundant species, S.triophthalma and S.cecilia,showed a clear dependence of b on the observed chlorophyll aconcentrations. Maximum reproductive rates ({small tilde}0.015h^–1) were attained only at relatively high phytoplanktondensities within a bloom of Heterocapsa triquetra where thechlorophyll a concentrations exceeded 10 µg l^–1.Estimates of secondary production suggest that Synchaeta spp.may contribute to the trophic flow of carbon in this systemwith a significance at least similar to that of the planktoniccopepods.     

The response of tomato roots (Lycopersicon esculentum Mill.) to iron deficiency stress: alterations in the pattern of protein synthesis

Dicotyledon plants adapt to iron (Fe) deficiency through a seriesof reactions that increase the ability of the plant to assimilateFe and to increase the efficiency of Fe utilization. In an attemptto gain an insight into these adaptive processes, the specificchanges in protein synthesis associated with the onset of theFe deficiency response in tomato roots (Lycopersicon esculentumMill cv. Rutgers) have been investigated. Roots were grown underFe—sufficient and —deficient conditions, and thepattern of protein synthesis was analysed by in vitro translationof root mRNA and by in vivo labelling of root proteins. Polypeptideswere resolved by two—dimensional polyacrylamide gel elec—trophoresis.Seven polypeptides were identified by in vitro translation,whose synthesis was significantly increased during Fe deficiency.The increase was probably specific to Fe deficiency in thatthe polypep—tide synthesis was not increased during phosphatedeficiency stress, was less prominent following prolonged Fedeficiency and was decreased following re—supply of Feto the hydroponic medium. The pattern of in vitro translation of mRNA isolated from Fe—deficientroots was compared to the results obtainedin vivo followingradiolabelling of proteins. In these analyses, eight polypeptideswere identified, tentatively including the seven polypeptidespreviously identified by in vitro translation. All polypeptideswere characterized with regard to molecular mass and pl andtheir localization in the cell, whether being membrane boundor soluble. It is suggested that members of this group of polypeptidesare involved in the response of the root to Fe deficiency: althoughtheir functions remain to be identified. Key words: In vitro protein synthesis, iron, iron deficiency, root, 2-dimensional PAGE     

PROMOTING ACTIVITY OF TERPENIC LACTONES IN PHASEOLUS ROOTING AND THEIR REACTIVITY TOWARD CYSTEINE

Sesquiterpenoids which contain exomethylenes conjugated to -lactonecarbonyl-heliangine, helianginol, pyrethrosin and cyclopyrethrosinacetate—promoted the adventitious root formation on hypocotylsof cuttings taken from light-grown (1900 lux) 6-day old Phaseolusmungo seedlings, but their derivatives in which the methylene-lactone systems are reduced-to the saturated lactones—dihydroheliangine,hexahydro-heliangine, dihydrohelianginol, dihydrocyclopyrethrosinacetate and tetrahydrocyclopyrethrosin acetate—showedno effect on the root formation. As far as the present experiment is concerned, every substancewhich showed promoting activity in Phaseolus rooting reactedwith cysteine and formed an adduct, but any of the substancesincapable of promoting root formation did not react with cysteine.The correlation of promoting activity of terpenic lactones inPhaseolus rooting with their reactivity toward SH groups maythus be demonstrated. ¹ Contribution No. 14 from the Botanical Gardens, Faculty ofScience, University of Tokyo, Tokyo.     

Some Complicating Factors in the Study of the Calcium Metabolism of Teleosts

A number of factors which may complicate studies dealing withthe calcium metabolism of teleosts are pointed out. Thus, lowenvironmental levels of calcium markedly affect both gill RNAand sodium metabolism in dilute sea water, and the absence ofmonovalent cations affects calcium metabolism in dilute environments.Hypophysectomy is not always followed by the disappearance ofcortisol from the plasma, and prolactin—which is requiredfor the survival of either hypophysectomized or intact animalsin a divalent ion-free environment—stimulates cortisolproduction. It is also pointed out that Fundulus kansae can be maintainedin divalent ion-free environments for long periods, and thatcalcium is mobilized from internal reservoirs such as acellularbone and scales. Failure to survive in such environments isdue to the regulatory effect of environmental calcium on sodiummetabolism rather than on calcium metabolism per se. Finally,it is pointed out that while one parameter of calcium metabolism—suchas plasma calcium levels in male fish—may remain constantthroughtout the year, other parameters such as calcium uptakemay vary considerably. Seasonal differencees in responses tomammalian calcitonin have also been noted.     

Growth and Dry-weight Distribution in Callistephus chinensis as Influenced by Lighting Treatment

Plants of two cultivars of Callistephus chinensis (Queen ofthe Market and Johannistag) were grown in 8 h of daylight perday with one of the following treatments given during the 16h dark period: (a) darkness—‘uninterrupted night’,(b) I h of light in the middle of the dark period—a ‘nightbreak’, (c) I min of light in every hour of the dark period—‘cycliclighting’, (d) light throughout—‘continuouslight’. The plants receiving uninterrupted dark periods remained compactand rosetted in habit with small leaves, while leaf expansion,stem extension, and flower initiation were promoted in all threeillumination treatments (b, c, d). Although these three treatmentsproduced similar increases in leaf area, continuous light wasthe most effective for the promotion of both stem growth andflower initiation while cyclic lighting was generally more effectivethan a I-h night break. Continuous light also caused more dry matter to be divertedto stems at any given vegetative dry weight and it was shownthat the stem weight ratio of both varieties was correlatedwith stem length.