Isolation of the ALG6 locus of Saccharomyces cerevisiae required for glucosylation in the N-linked glycosylation pathway

N-Linked protein glycosylation in most eukaryotic cells initiateswith the transfer of the oligosaccharide Glc₃Man₉GlcNAc₂ fromthe lipid carrier dolichyl pyrophosphate to selected asparagineresidues. In the yeast Saccharomyces cerevisiae, alg mutationswhich affect the assembly of the lipid-linked oligosaccharideat the membrane of the endoplasmic reticulum result in the accumulationof lipid-linked oligosaccharide intermediates and a hypoglycosylationof proteins. Exploiting the synthetic growth defect of alg mutationsin combination with mutations affecting oligosaccharyl transferaseactivity (Stagljar et al., 1994), we have isolated the ALG6locus. alg6 mutants accumulate lipid-linked Man₉GlcNAc₂, suggestingthat this locus encodes an endoplasmic glucosyltransferase.Alg6p has sequence similarity to Alg8p, a protein required forglucosylation of Glc₁Man⁹GlcNAc². Saccharomyces cerevisiae endoplasmic reticulum glycosyltransferase dolichol     

Homonojirimycin and N-methyl-homonojirimycin inhibit N-Iinked oligosaccharide processing

Homonojirimycin (HNJ) and N-methylhomonojirimycin (MHNJ) weretested as inhibitors of the purified glycopro-tein processingenzymes, glucosidase I and glucosidase II. MHNJ was a reasonablygood inhibitor of glucosidase I (K₁ = 1 x 10^–6 M) andwas about three times as effective on this enzyme as was HNJ.On the other hand, HNJ inhibited glucosidase II with a K₁ ofabout 1 x 10^x6 M, whereas MHNJ was three times less effective(K₁ = 3 x 10^–5 M). However, the butyl derivative of HNJhad very low activity toward these two processing glucosidases.HNJ and its methyl derivative were also tested in vivo usinginfluenza virus-infected MDCK cells, and measuring the inhibitionof N-linked oligosaccharide processing of the viral envelopeglycoproteins. With 100 µg/ml of MHNJ in the medium, essentiallyall of the N-linked oligosaccharide chains of the virus wereof the "high-mannose" type with the major structure being characterizedas Glc₃Man₉(GlcNAc)₂. Similar results were obtained with HNJalthough this compound was less effective in vivo as well asin vitro. These results are in keeping with these inhibitorsbeing effective at the glucosidase I step. Both inhibitors werealso tested in MDCK cell cultures to determine whether theyaffected the in vivo synthesis of proteins, or of lipid-linkedsaccharides. In contrast to deoxynojirimycin, which has beenreported to inhibit the formation of lipid-linked saccharides,no effects were seen on either the incorporation of mannoseinto lipid-linked saccharides or the incorporation of leucineinto protein. glucosidase lipid influenza virus oligosaccharide     

Molecular dynamics simulations of high-mannose oligosaccharides

Conformations of several high-mannose-type oligosaccharidesthat are generated during the biosynthetic degradation of Man₉GlcNAc₂to Man₅GlcNAc₂ have been studied by molecular dynamics (MD).Simulations were performed on NCI-FCRDC's Cray Y-MP 8D/8128supercomputer using Biosym's CVFF force field for 1000 Ps withdifferent initial conformations. The conformations of the two1,3- and the two 1,6-linkages in each oligomannose were different,suggesting that deriving oligosaccharide conformations basedon the conformational preferences of the constituent disaccharidefragments will not always yield correct results. Unlike otheroligomannoses, Man₉GlcNAc₂ appears to take more than one distinctconformation around the core 1,6-linkage. These various conformationsmay play an important role in determining the processing pathways.Using the data on the preferred conformations of these oligomannosesand the available experimental results, possible pathways forprocessing Man₉GlcNAc₂ to Man₅GlcNAc₂ by 1,2-linkage-specificmannosidases have been proposed. Conformational analysis ofMan₅GlcNAc₂ indicates that the addition of ß1,2-GlcNActo the 1,3-linked core mannose, besides serving as a prerequisitefor mannosidase II action as suggested earlier, may also preventthe removal of 1,3-mannose. The MD simulations also suggestthat the processing of the precursor oligosaccharide duringAsn-linked complex and hybrid glycan biosynthesis proceeds ina well-defined pathway involving more than one 1,2-linkage-specificmannosidase. Knowledge of the conformation of the processingintermediates obtained from the present study can be used todesign highly specific substrate analogues to inhibit a particularmannosidase, thereby blocking one processing pathway withoutinterfering with the others. carbohydrates conformation glycosidase inhibitors mannosidase oligosaccharide processing     

Structure of the N-linked oligosaccharide of the main diagnostic antigen of the pathogenic fungus Paracoccidiodes brasiliensis

The major diagnostic antigen of Paracoccidioides brasiliensisis the exocellularly secreted 43,000 Da glycoprotein (gp43)which contains a single N-linked oligosaccharide chain. Thisoligosaccharide, although poorly immunogenic in man, is responsiblefor the cross-reactivity of the gp43 with sera from patientswith histoplasmosis, and may have a role in fungal virulence.It contains a neutral highmannose core (Man₇GlcNAc₂) to whicha (1     

Cloning and characterization of the ALG3 gene of Saccharomyces cerevisiae

The Saccharomyces cerevisiae alg3-1 mutant is descilbed as defectivein the biosynthesis of dolichol-linked oligosaccharides (Huffakerand Robbins, Proc. Natl. Acad. Sci. USA, 80, 7466–7470,1983). Man₅GlcNAc₂-PP-Dol accumulates in alg3 cells and EndoH resistant carbohydrates are transferred to protein by theoligosaccharyltransferase complex. In this study, we describethe cloning of the ALG3 locus by complementation of the temperaturesensitive growth defect of the alg3 stt3 double mutant. Theisolated ALG3 gene complements both the defect in the biosynthesisof lipidlinked oligosaccharides of the alg3-mutant and the underglycosylationof secretory proteins. The inactivation of the nonessentialALG3 gene results in the accumulation of lipid-linked Man₅GlcNAc₂and protein-bound carbohydrates which are completely Endo Hresistant. The ALG3 locus encodes a potential ER-transmembraneprotein of 458 amino acids (53 kDa) with a C-terminal KKXX-retrievalsequence. lipid-linked oligosaccharide N-glycosylation synthetic lethality     

Comparison of NMR and molecular modeling results for a rigid and a flexible oligosaccharide

Three-bond heteronuclear coupling constants (³J_CH) are extremelyuseful in describing flexible models for oligosaccharides. Weshow that antiphase methods for measuring ³J_CH in oligosaccharideshave limited reliability but that the coupling constants canbe reliably measured in natural abundance by quantitative J-correlationmethods. Interpretation of ³J_CH data for a pentasaccharide (lacto-N-fuco-pentaose2) from human milk are consistent with a rigid model for theLewis^a trisaccharide epitope but for an antigenic tetrasaccharidefragment from the cell wall polysaccharide of viridans streptococci,³J_CH data imply a considerably more flexible model. NuclearOverhauser effect (NOE) data are reported for a heptasacchariderepeating unit isolated from the cell wall polysaccharide ofStreptococcus gordonii 38. The results for a tetrasaccharidefragment are similar to data reported for the same fragmentin the cell wall polysaccharide from S.mitis 322. This resultimplies a similar conformation for the tetrasaccharide fragmentin the polysaccharide and in the heptasaccharide and also impliesthat anisotropy of motion is not significant in the interpretationof the nuclear Overhauser effects in the polysaccharide. Interpretationof the NOE results for the tetrasaccharide fragment, like the³J_CH data, implies a flexible model with three conformationsin fast exchange. The results of the two experimental techniquesare combined with molecular modeling results including moleculardynamics simulation to provide a clear delineation between flexibleand rigid oligosaccharide epitopes. The blood group Lewis^a trisaccharideantigenic determinant is highly restricted in its motions bysteric interactions while the antigenic tetrasaccharide fragmentof the S.gordonii 38 heptasaccharide is considerably more mobile.We propose that some branched oligosaccharides are relativelyrigid and some are flexible depending on subtle details of thelinkages. oligosaccharide conformation molecular dynamics     

Elongation of the N-glycans of fowl plague virus hemagglutinin expressed in Spodoptera frugiperda (Sf9) cells by coexpression of human {beta}1,2-N-acetylglucosaminyltransferase I

Spodoptera frugiperda (Sf9)-cells differ markedly in their proteinglycosylation capacities from vertebrate cells in that theyare not able to generate complex type oligosaccharide side chains.In order to improve the oligosaccha ride processing propertiesof these cells we have used baculovirus vectors for expressionof human (ß1,2-N-acetylglucosaminyltransferase I (hGNT-I),the enzyme catalysing the crucial step in the pathway leadingto complex type N-glycans in vertebrate cells. One vector (Bac/GNT)was designed to express unmodified GNT-I protein, the secondvector (Bac/tagGNT) to express GNT-I protein with a tag epitopefused to its N-terminus. In Sf9-cells infected with Bac/tagGNT-virusa protein of about 50 kDa representing hGNT-I was detected withan antiserum directed against the tag epitope. HGNT-I activitywas increased at least threefold in lysates of infected cellswhen N-acetylglucosamine (GlcNAc)-free ovalbumine was used assubstrate. To monitor hGNT-I activity in intact Sf9-cells, theglycosylation of coexpressed fowl plague virus hemagglutinin(HA) was investigated employing a galactosylation assay andchromatographic analysis of isolated HA N-glycans. Coexpressionof hGNT-I resulted in an at least fourfold increase of HA carryingterminal GlcNAc-residues. The only structure detectable in thisfraction was GlcNAcMan₃GlcNAc₂. These results show that hGNT-Iis functionally active in Sf9-cells and that the N-glycans ofproteins expressed in the baculovirus/insect cell system areelongated by coexpression of glycosyltransferases of vertebrateorigin. Complete complex type oligosaccharide side chains werenot observed when hGNT-I was overexpressed, thus supportingthe concept that Sf9-cells do not contain glycosyltransferasesacting after hGNT-I. ß1,2-N-acetylglucosaminyltransferase I baculovirus expression of recombinant protiens N-glycosylation in Sf9-cells     

Characterization of lipophosphoglycan from a Ricin-resistant mutant of Leishmania major

One of the virulence factors of the protozoan parasite Leishmaniamajor is the surface glycoconjugate, lipophosphoglycan (LPG).A Ricin-resistant mutant of L.major was generated and characterisedwith respect to its virulence in mice and the structure andexpression of LPG. The LPG from this mutant (1F6-B5) retainedthe tripartite structure of wild-type LPG, comprising a glycosylphosphatidylinositol(GPI) anchor linked to a phosphorylated disaccharide backboneterminating in a nonreducing neutral oligosaccharide cap. Thestructure of the GPI anchor and the major capping oligosaccharidewere identical to wild-type LPG. However, there were variationsin the number of phosphorylated repeats (PO₄-6Gal(ß1-4)Man(     

Oligosaccharide profiles of HIV-2 external envelope glycoprotein: dependence on host cells and virus isolates

The glycosylation pattern of the external envelope glycoproteinof human immunodeficiency virus type 2 (HIV-2) was studied independence on host cells and virus isolates. Strains HIV-2_ALT,HIV-2_ROD and HIV-2_D194, differing in their biological propertiesand in the amino acid sequences of their env genes, were propagatedin MOLT4, HUT78 and U937 cells, in human peripheral blood lymphocytesand monocytes/macrophages in the presence of [6-³] glucosamine.Radiolabelled viral glycoproteins were isolated from the cell-freesupernatants and digested with trypsin. Glycans were sequentiallyliberated by endo-ß-N-acetylglucosaminidase H andpeptide-N⁴-(N-acetyl-ß-glucosaminyl) asparagine amidaseF, and fractionated according to charge and size. Comparisonof the oligosaccharide profiles revealed that the envelope glycoproteinsof different virus isolates, propagated in the same host cells,yielded very similar glycan patterns, whereas cultivation ofan isolate in different host cells resulted in markedly divergentoligosaccharide maps. Variations concerned the proportion ofhigh-mannose-, hybrid- and complex-type substituents, as wellas the state of charge and structural parameters of the complex-typespecies. As a characteristic feature, complex-type glycans ofmacrophage-derived viral glycoprotein were almost exclusivelysubstituted by lactosamine repeats. Hence, glycosylation ofthe HIV-2 external envelope glycoprotein seems to be primarilygoverned by host cell-specific factors rather than by the aminoacid sequence of the corresponding polypeptide backbone. envelope glycoprotein glycosylation human immunodeficiency virus type 2     

Sulfated glycans on oral mucin as receptors for Helicobacter pylori

Helicobacter pylori is able to colonize gastric epithelia, causingchronic active gastritis, gastric and duodenal ulcers and presumablygastric malignancies. Attempts to identify the natural reservoirfor this microorganism other than the stomach have been unsuccessful.It is suspected that H.pylori can be transmitted orally, sincethe microorganism has been detected at various sites of theoral cavity. The aim of the present study was to determine whetherH.pylori can bind to salivary mucins, which in vivo coat theoral epithelia, and characterize further the interaction. Bindingof salivary mucins and of synthetic oligosaccharides was studiedIn ELISA and immunoblotting, using specific mono-and polyclonalantibodies, and synthetic neoglycoconjugates. H.pylori boundmost avidly to a highly sulfated sub-population of high molecularweight salivary mucins, secreted from the palatine salivaryglands, and with less avidity to mucin species secreted by thesublingual and submandibular salivary glands, which are lesssulfated. Binding was strongly enhanced upon decreasing pH from6.0 to 5.0. Using synthetic polyacrylamide coupled oligosaccharidesit was found that SO₃-3-Gal and the SO₃-3-Lewis^a blood groupantigen bound to H.pylori. In contrast, binding of sialylatedLewis^a and Lewis^b antigens was much weaker. This study indicatesthat sulfated oligosaccharides on salivary mucins may providereceptor structures for adhesion of H.pylori to oral surfaces. H.pylori saliva sulfomucin nickel oligosaccharide     

Dodecyl-{beta}-D-maltoside as a substrate for glucosyl and xylosyl transfer by glycogenin

Glycogenin is the core protein of glycogen proteoglycan andis, at the same time, a self-glucosylating enzyme which catalysesearly glucosyl transfer steps in the biosynthesis of glycogen.In the course of this process, glycogenin is glucosylated progressivelyuntil an oligosaccharide containing 8–11 glucose residueshas been formed. Although glycogenin, under physiological conditions,is both enzyme and acceptor in the glucosyl transfer reactions,it is also capable of utilizing p-nitrophenyl-linked malto-oligosaccharidesas exogenous acceptors. In view of the potential usefulnessof exogenous acceptors in the study of the mechanism of theglycogenin reaction, we have expanded the search for such compoundsand report here that several alkyl glucosides and alkyl maltosidesmay serve as acceptors in glucosyl transfer by beef kidney glycogenin.Dodecyl-ß-D-maltoside (K_m {small tilde}100 µM)was the most effective acceptor among the compounds tested andyielded 30 times as much product as p-nitrophenyl-     

N-Glycosylation in trypanosomatid protozoa

Trypanosomatid protozoa are parasites of considerable medicaland economic importance in developing countries. The pathwayleading to N-glycosylation in these micro-organisms is characterizedby the following features: (i) dolichols are composed of only10–13 isoprene units; (ii) oligosaccharides transferredin N-glycosylation have the compositions Man_{(6, 7, 9)}GlcNAc₂,depending on the species; (iii) trypanosomatids are unable tosynthesize dolichol-P-Glc and, in addition, some species lackcertain dolichol-P-Man-dependent mannosyltransferases; (iv)the oligosaccharyl-transferase does not require the presenceof glucose units in the oligosaccharide in order to catalysean efficient transfer reaction; (v) trypanosomatids have a glucosidaseII-like enzyme, but lack glucosidase I; (vi) glucosidase IIis required for deglucosylation of oligosaccharides glucosylatedby the UDP-Glc: glycoprotein glucosyltransferase, an activityfirst detected in those parasites; (vii) the structures of polymannose-typecompounds in these protozoa have no significant differenceswith those of their mammallan counterparts except for the presence,in certain species, of oligosaccharides having galactofuranoseunits linked to external mannose residues; (viii) biantennarycomplex-type oligosaccharides having in some cases terminal     

Simulation and Prediction of Plant Phenology for Five Crops Based on PhotoperiodxTemperature Interaction

This paper presents a plant phenological model based on genotypextemperaturexphotoperiodinteraction (GPTmodel). In the model, rate of development towardsa specified stage (e.g. flowering) for a given genotype is composedof three components: the genotype's maximum rate of development;any delay due to a non-optimal temperature; and any delay dueto a photoperiod response. It is assumed that development tothe specified stage is an autonomous process established bymost, if not all, genes other than the vernalization genes andthe photoperiod genes; and that this autonomous process is delayedby any activity of the photoperiod genes. Since all physiologicalprocesses are modulated by temperature, any photoperiod responseis inevitably a photoperiodxtemperature interaction. This interactionis simulated by assuming that the photoperiod gene activityoccurs only beyond a critical photoperiod (P_c) and is enlargedby temperature above a base temperature (T_bp) that allows thephotoperiod gene activity. The model is written asR=1/D_b-S_t(T-T_opt)²-S_p(T-T_bp)|P-P_c|, whereRis the expected rate of development to the specifiedstage under any combination of temperature (T) and photoperiod(P). The other model parameters are:S_p, the sensitivity to adelaying photoperiod;T_opt, the optimum temperature for developmentin the absence of the photoperiod response;S_t, the sensitivityto a non-optimum temperature; andD_b, the basic duration to thespecified stage (or intrinsic earliness), the inverse of whichis the maximum rate of development.D_bis observable only ifT=T_optandsimultaneouslyP     

Potential Dry Matter and Water Import Rates in the Tomato Fruit in Relationship to Fruit Size

The tomato fruit was compared to a sphere with a radius R. Radialgrowth rates in the fruit (F_IW and F_ID) due to water importor to dry matter import, respectively, which are also the waterimport rate or dry matter import rate per unit surface areaof fruit, were calculated from two sets of published results.This data referred to fruits which swelled in such a way thatthe availability of assimilates had little effect on growth.Two varieties differentiated the two series of results and inone series, three trials were differentiated by the salinityof the nutrient solution. In all trials, it was found that F_IW and F_ID decreased whenR increased. Two phases were observed for F_IW: after a firstphase, F_IW decreased more quickly and almost linearly when Rincreased. F_ID was constant or decreased with respect to R.Except at the beginning of growth at the greatest salinity,there were clearly linear regressions between F_IW and F_ID suchas F_ID = aF_IW-b; where b was lower with higher salinity. Thechanges of the concentration of imported dry matter (F_ID/F_IWwere examined in terms of R and F_IW/R. The mechanisms controllingthe changes in F_IW and F_ID were discussed. The results suggestedfruit radius was an important parameter of these mechanisms.Thus, water import rate and dry matter import rate could eachbe considered to be the product of two factors: fruit surfacearea, which is directly dependent on fruit radius, and waterimport rate or dry matter import rate per unit of fruit surfacearea.Copyright 1993, 1999 Academic Press Dry matter, fruit growth, logistic model, sink size, tomato, water transfer     

Effects of Photoperiod and Maturity Genes on Plant Growth, Partitioning, Radiation Use Efficiency, and Yield in Soyabean [Glycine max(L.) Merrill] 'Clark'

Plants of four isolines of soyabean [Glycine max(L.) Merrill]‘Clark’, viz‘L71-920’ (maturity genecomplemente₁e₂e₃ ), ‘L80-5914’ (E₁e₂e₃), ‘Clark’(e₁E₂E₃), and ‘L65-3366’ (E₁E₂E₃), were grown inshort (12.25 h d ^{- 1}natural light) and long days (12.25 h d^{- 1}natural light supplemented with 2.75 h d ^{- 1}low-irradianceartificial light) from first flowering to maturity in a polythenetunnel maintained at 30/24°C (day/night). Whereas therewere few differences among the isolines grown in short days,in long days the dominant alleles increased crop duration, biomassand seed yield substantially. Increases in biological and economicyield were not solely a consequence of longer crop duration:the dominant alleles also increased crop growth rate and radiationuse efficiency in long days (from 1.3 g MJ ^{- 1}total radiationine₁e₂e₃ to 2.8 g MJ ^{- 1}inE₁E₂E₃ ). Greater radiation use efficiencyresulted from a relatively longer leaf area duration, betterdistribution and orientation of a larger mass of leaves withinthe canopy, and smaller partitioning of assimilates to reproductivestructures. The work reveals the substantial effects of thethree lociE₁ / e₁, E₂/ e₂and E₃/e₃ on the response of plantgrowth, as well as development, to environment. Their relevanceto crop adaptation is discussed. Copyright 2000 Annals of BotanyCompany Glycine max(L.) Merrill, soyabean, maturity genes, flowering, phenology, growth, yield     

Partitioning of Transfer and Carboxylation Components of Intracellular Resistance to Photosynthetic CO2 Fixation: A Critical Analysis of the Methods Used

The partitioning of intracellular resistance to CO₂ transferin two components: mesophyll (r_m.) and carboxylation (r_x) resistances,is based upon the combination of two sub-models: one encompassingtransfer processes and the second, the carboxylation system.All the determinations derived from this method yield a highr_m/r_x ratio. it is demonstrated in low oxygen conditions, whateverthe model used, that this conclusion is highly-dependent uponthe form of the equation used at the carboxylation level. Thepossible influence of O₂ concentration on the r_m/r_x ratio isdiscussed. Starting from a Rabinowitch model (rectangular hyperbola), someof the conditions necessary to yield lower r_m/r_x ratio are considered.It is shown that the most relevant factor when modelling theCalvin cycle is the number of limiting pools (enzymes or cofactors)rather than the number of limiting reactions.     

Growth of Plants in Different Oxygen Concentrations

Dwarf french beans (Phaseolus vulgaris var. Canadian Wonder)were grown in chambers at 25?C with the roots aerated at 20per cent oxygen and tops variously maintained at: T₁ O₂ 0.21;CO₂ 270?10^–6: T₂; O₂ 0.05, CO₂, CO₂ 270?10^–6: T₃;O₂ 0.21; CO₂ 550?10^–6. Experiment 1 (T₁ and T₂) lasted2 weeks: Experiment 2 (T₁ T₂ and T₃) only one week. Hourly estimatesof CO₂ uptake were made by gas analysis and weekly estimatesof fresh weight, dry matter in tops and roots, and leaf area,by sampling. Light intensity was 80 W m^–2 of photosyntheticallyactive radiation. An attempt was made to explain the results in terms of a simplelight absorption model such that where dV/dt is the rate of CO₂ uptake per plant, ßis the photosynthetic efficiency, I₀ is the incident light intensity,f is the fraction of incident light absorbed by unit leaf layerand L is the leaf area index. The analysis showed that ß(T₂)was at least double ß(T₁), whilst f(T₂) was smallerthan f(T₁) at a given leaf area. The results also required thatthroughout the period of the experiment, fL(T₁)=fL(T₂) at anygiven time, i.e. the treatment with the larger leaf area (T₂)has the smaller value of f, and therefore intercepts less lightper unit leaf area. This could be advantageous for plant growth,but requires further experiments. The photosynthetic rates per unit leaf are about 40 per centgreater in T₂ than T₁. Over the relatively short period of the experiment the resultsare adequately described by U=btⁿ, where U is the accumulatedcarbon dioxide uptake, b is related to the photosynthetic efficiency(different for the differing treatments), and n is a constant(similar for all treatments). This relationship with time isbelieved to be a relationship with accumulated radiation, forthe light was constant throughout the experiments. Comparisons of carbon fixed (measured gas uptake) and dry matteraccumulation (sampling) show great scatter with an average valueof 0.43. The first week's results were generally smaller thanthis value and the second week's greater. Energy fixation as a fraction of photosynthetically active radiationon the ground area covered by the plants ranged from 3.5 to10 per cent. The results from treatment T₃ were similar to T₂ suggestingthat increasing CO₂ concentration decreases the growth inhibitionat 21 per cent O₂.     

Characterization of Photothermal Flowering Responses in Maturity Isolines of Soyabean [Glycine max (L.) Merrill] cv. Clark

All eight isolines of three maturity genes (E₁/e₁, E₂ /e₂, andE₃ /e₃) of soyabean [Glycine max (L.) Merrill] cv. Clark weregrown in widely different combinations of photoperiod and temperature.Under the more inductive conditions, i.e. in a warm mean temperature(30°C) when daylengths were less than the critical value(i.e. less than about 13 h), the isolines flowered at similartimes (23-24 d). The responses of all isolines to temperaturewere also similar, if not identical. Increase in daylength abovethe critical photoperiod progressively delayed flowering untilthe time taken to flower (f) reached a maximum at the ceilingphotoperiod. The relations between the rate of progress towardsflowering (1/f) and photoperiod (between the critical and ceilingvalues) were linear. The coefficient characterizing the slopeof the response (photoperiod sensitivity) varied amongst theisolines. These responses could be grouped into three categoriesof increasing sensitivity: (1) least sensitive, e₁e₂e₃ , e₁E₂e₃, e₁e₂E₃ ; (2) intermediate, E₁e₂e₃ , e₁E₂E₃ ; and (3) mostsensitive, E₁E₂e₃, E₁e₂E₃ , E₁E₂E₃ . Thus, in the Clark cultivargenetic background, E₁ induces greater photoperiod sensitivitybut neither E₂ nor E₃ on their own have any effect. However,both E₂ and E₃ together induce photoperiod sensitivity comparableto that induced by E₁ alone. Furthermore, in addition to thisepistasis, either E₂ or E₃ has considerable epistatic effecton E₁, further increasing photoperiod sensitivity. The effectsof these genes and their epistasis were also reflected in theextent of the maximum delays to flowering which occur when theceiling photoperiod is exceeded, and also possibly in earlinessin circumstances when photoperiods were below the critical value.Copyright1994, 1999 Academic Press Glycine max (L.) Merrill, soyabean, maturity genes, flowering, photoperiod, temperature