srb: a Bacillus subtilis Gene Encoding a Homologue of the {alpha}-Subunit of the Mammalian Signal Recognition Particle Receptor

We cloned a Bacillus subtilis gene (srb) encoding a homologueof the mammalian signal recognition particle receptor -subunit(SR). The gene is 987 bp in length and encodes a 329-amino acidprotein. The deduced amino acid sequence of the protein shared26.6, 36.2 and 49.7% identity with those of mammalian SR, archaebacterialDP and Escherichia coli FtsY, respectively. The protein containsthree conserved GTP-binding elements like the other three SRPreceptor proteins, though the N-terminal portion of the putativeB. subtilis protein was shorter than the others. Secondary structureprediction showed that an amphipathic -helix is positioned inthe N-terminal region. A defect in srb inhibited cell growthand protein translocation.     

Genotypic Differences in Leaf Water Relations between Brassica juncea and B. napus

Various kinds of measurement of tissue water status were madeseveral times during water stress and recovery in Brassica juncea(cv Canadian Black) and B napus (cv Drakkar) Unstressed plantsof the two species had similar leaf water potentials (_w), solute(_s) and turgor potentials (_p) Values of relative water content(RWC) and the slope of the linear relationship between _p andRWC (_p/RWC) were greater in B napus than in B juncea Statistical correlations of pooled data for the watered andstressed treatments differentiated the relationships among RWC,_w and its components in the two species The major statisticaldifference was that _p/RWC was related to RWC in B napus andto _w and _s in B juncea A decline in _p/RWC with decreasing _sin B juncea may be a mechanism for maintaining _p at low soilwater potentials through maintenance of more elastic cell walls. Brassica juncea, Brassica napus, osmotic adjustment, tissue elasticity, water relations     

Effects of Priming and Endosperm Integrity on Seed Germination Rates of Tomato Genotypes: II. GERMINATION AT REDUCED WATER POTENTIAL

Seed germination rates (GR =inverse of time to germination)are sensitive to genetic, environmental, and physiological factors.We have compared the GR of tomato (Lycopersicon esculentum Mill.)seeds of cultivar T5 to those of rapidly germinating L. esculentumgenotypes PI 341988 and PI 120256 over a range of water potential(). The influence of seed priming treatments and removal ofthe endosperm/testa cap enclosing the radicle tip on germinationat reduced were also assessed. Germination time-courses atdifferent 's were analysed according to a model that identifieda base, or minimum, allowing germination of a specific percentage(g) of the seed population (_b(g)), and a ‘hydrotime constant’(_H) indicating the rate of progress toward germination per MPa.h.The distribution of _b(g) determined by probit analysis was characterizedby a mean base (_b) and the standard deviation in _b among seeds(_b). The three derived parameters, _b, _b) and _H, were sufficientto predict the time-courses of germination of intact seeds atany . A normalized time-scale for comparing germination responsesto reduced is introduced. The time to germination at any (t_g())can be normalized to be equivalent to that observed in water(t_g(0)) according to the equation t_g(0)=[l–(/_b(g))]t_g().PI 341988 seeds were more tolerant of reduced and had a morerapid GR than T5 seeds due to both a lower _b and a smaller _H.The rapid germination of PI 120256, on the other hand, couldbe attributed entirely to a smaller _H. Seed priming (6 d in–1.2 MPa polyethylene glycol 8000 solution at 20 ?C followedby drying) increased GR at all >_b(g), but did not lower theminimum allowing germination; i.e. priming reduced _H withoutlowering _b. Removing the endosperm/testa cap (cut seeds) markedlyincreased GR and lowered the mean required to inhibit germinationby 0.7 to 0.9 MPa. However, this resulted primarily from downwardadjustment in _b during the incubation of cut seeds at low inthe test solutions. The difference in _b between intact and cutseeds incubated at high was much less (0.l MPa), indicatingthat at the time of radicle protrusion, the endosperm had weakenedto the point where it constituted only a small mechanical barrier.In the intact seed, endosperm weakening and the downward adjustmentin embryo _b ceased at < –0.6 MPa, while the reductionin _H associated with priming proceeded down to at least –1.2MPa. Based on these data and on the pressure required to pushthe embryos from the seeds at various times after imbibition,it appears that the primary effect of priming was to shortenthe time required for final endosperm weakening to occur. However,as priming increased GR even in cut seeds, priming effects onthe embryo may control the rate of endosperm weakening. Key words: tomato, Lycopersicon esculentum Mill., water potential, germination rate, seed priming, genetic variation     

Physiological Responses to Drought of Lolium perenne L.: Measurement of, and Genetic Variation in, Water Potential, Solute Potential, Elasticity and Cell Hydration

Thomas, H. 1987. Physiological responses to drought of Loliumperenne L.: Measurement of, and genetic variation in, waterpotential, solute potential, elasticity and cell hydration.—J.exp. Bot. 38: 115–125. Clonally-replicated genotypes of Loiium perenne L. were grownin a controlled environment. Leaf water potential (_w) osmoticpotential (_s), turgor potential (_p = _w – _s), elasticity(E), leaf hydration (g water per g dry matter, H) and numberof green leaves per tiller (N_GL) were measured before and duringa 42 d drought treatment. A simplified method of estimating E (at _w < 1?0 MPa) usingonly six measurements was developed to permit a measurementrate of 8 leaves per hour. Measurement errors in all characterswere 3% or less. During drought, _w and _s (at _w = 0?5 MPa) decreased significantly,_p and E increased significantly, and H decreased slightly. Plantsize during drought was negatively correlated with _s, and ^Hand positively correlated with _p, osmotic adjustment, E andN_GL. Measurements made on the genotypes before draughting didnot give a reliable indication of their physiological conditionafter adaptation to drought. Genetically controlled variation (‘broad sense heritability’)of drought-adapted plants for E was 15%, _w 23%, _s, 34%, _p, 35%,H 34% and N_GL 64%. The possibilities for, and effectivenessof, divergent selection of genotypes with high and low expressionof the characters are discussed. Key words: Water relations, Lolium, genetic variation     

Sugar-responsive gene expression, invertase activity, and senescence in aborting maize ovaries at low water potentials

• Background and Aims Ovary abortion can occur in maize(Zea mays) if water deficits lower the water potential (_w) sufficientlyto inhibit photosynthesis around the time of pollination. Theabortion decreases kernel number. The present work exploredthe activity of ovary acid invertases and their genes, togetherwith other genes for sucrose-processing enzymes, when this kindof abortion occurred. Cytological evidence suggested that senescencemay have been initiated after 2 or 3 d of low _w, and the expressionof some likely senescence genes was also determined. • Methods Ovary abortion was assessed at kernel maturity.Acid invertase activities were localized in vivo and in situ.Time courses for mRNA abundance were measured with real timePCR. Sucrose was fed to the stems to vary the sugar flux. • Key Results Many kernels developed in controls but mostaborted when _w became low. Ovary invertase was active in controlsbut severely inhibited at low _w for cell wall-bound forms invivo and soluble forms in situ. All ovary genes for sucroseprocessing enzymes were rapidly down-regulated at low _w exceptfor a gene for invertase inhibitor peptide that appeared tobe constitutively expressed. Some ovary genes for senescencewere subsequently up-regulated (RIP2 and PLD1). In some genes,these regulatory changes were reversed by feeding sucrose tothe stems. Abortion was partially prevented by feeding sucrose. • Conclusions A general response to low _w in maize ovarieswas an early down-regulation of genes for sucrose processingenzymes followed by up-regulation of some genes involved insenescence. Because some of these genes were sucrose responsive,the partial prevention of abortion with sucrose feeding mayhave been caused in part by the differential sugar-responsivenessof these genes. The late up-regulation of senescence genes mayhave caused the irreversibility of abortion.     

Analysis of Pressure-Volume Curves of Leaves of Rosa hybrida cv. Sonia

By analysing the relationship between inverse water potential(^–1), and relative water content (RWC) measured on leavesof roses (Rosa hybrida cv. Sonia), grown soilless, it was foundthat a non-linear (NL) model was better suited than a linearmodel to reproduce values observed in the non-turgid region.To explain this apparent curvature, it is assumed that a reductionof the non-osmotic water fraction (A_p) takes place when decreases.Osmotic potentials () measured on fresh and frozen leaf discstend to support this hypothesis. A method for exploiting PVcurves, which takes into account the variation of A_p, is described.It delivers values for the turgor pressure (_p), the relativeosmotic water content, and the mean bulk volumetric elasticitycoefficient, lower than those given by the linear model. Onthe other hand, it gives higher estimates for A_p and for . Whenapplying the traditional model to obtain estimates for waterrelations characteristics of rose leaves, and comparing resultsfrom two distinct salinity treatments (electrical conductivitiesof 1·8 mS cm^–1 and 3·8 mS cm^–1, respectively),one deduces a significant reduction of at turgor-loss in thehigh salinity treatment. The NL method is, in addition, ablesimultaneously to reveal a reduction of and a significant increasein _p at RWC=100% this proves that soilless–grown roseplants are able to osmoregulate when subjected to a constantand relatively high degree of salinity. Key words: Apoplastic water, non-linear regression, pressure-volume curves, tissue-water relations     

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     

Integrated likelihood functions for non-Bayesian inference

Consider a model with parameter = (, ), where is the parameterof interest, and let L(, ) denote the likelihood function. Oneapproach to likelihood inference for is to use an integratedlikelihood function, in which is eliminated from L(, ) by integratingwith respect to a density function (|). The goal of this paperis to consider the problem of selecting (|) so that the resultingintegrated likelihood function is useful for non-Bayesian likelihoodinference. The desirable properties of an integrated likelihoodfunction are analyzed and these suggest that (|) should be chosenby finding a nuisance parameter that is unrelated to and thentaking the prior density for to be independent of . Such anunrelated parameter is constructed and the resulting integratedlikelihood is shown to be closely related to the modified profilelikelihood.     

Abscisic Acid and Water Relations of Rice (Oryza sativa L.): Sequential Responses to Water Stress in the Leaf

Water stress was imposed by withholding water at an early vegetativestage from plants of two rice cultivars (IR20 and 63–83)grown in pots. As stress intensified the following sequenceof responses of the leaves was observed: (i) rise in abscisicacid (ABA) content, (ii) closure of stomata, (iii) initiationof leaf rolling. In both cultivars, turgor (_p) declined linearly with total waterpotential () of the leaf. Bulk leaf ABA content increased linearlyas _p declined, and attained twice the control (unstressed) levelfollowing a reduction in _p of about 0.12 MPa. Stomatal conductance exhibited a sigmoidal relationship to _p,declining abruptly when a particular ‘critical’_p was reached (threshold response). The critical potentialsvaried considerably between experiments, but were closely correlatedwith control potentials and with the potentials at which ABAconcentration doubled relative to controls. Leaf rolling was initiated at s near to zero _p. Increases inthe ratio of adaxial to abaxial conductance were associatedwith rolling. Variations in the above responses could be accounted for byvariations in the rate of stress development, which in termsof reduction ranged from 0.38 to 0.86 MPa day^–1. Fastdrying rates resulted in: (a) reduced osmotic adjustment, (b)increased amounts of ABA in the leaf at a given level of or_p, (c) an increase in the ABA concentration present at 50 percent stomatal closure, and (d) initiation of leaf rolling ata higher . Oryza sativa L., rice, water stress, stomata, leaf rolling, abscisic acid     

Respiration and germination rates of tomato seeds at suboptimal temperatures and reduced water potentials

Suboptimal temperature (T) affects germination rates (reciprocalof time to radicle emergence) on a thermal time basis; thatis, the T in excess of a base or minimum temperature multipliedby the time to a given per cent germination [t_g) is a constant.Respiration rates are also sensitive to T, and proportionalrelationships are often found between respiration rates andgermination rates. Reduced water potential () delays seed germinationon a hydrotime basis (i.e. the in excess of a base water potentialmultiplied by t_g is a constant). It was tested whether respirationrates prior to radicle emergence vary in proportion to T and as expected from the thermal and hydrotime models. Respirationrates (C0₂ evolution) of cold-tolerant, rapidly germinating(PI 341988) and cold-sensitive, more slowly germinating (T5)tomato [Lycopersicon esculentum Mill.) seeds were evaluatedover a range of T and conditions. For both genotypes, respirationrates until the beginning of radicle emergence were relatedto T on a thermal time basis and increased approximately linearlywith above -2.0 MPa, consistent with the hydrotime model. Respirationrates were uniquely related to germination rates, regardlessof whether germination timing was affected by T, , or genotype.However, germination timing was unaffected when respirationrates were manipulated by varying 0₂ partial pressure. Thus,while both germination and respiration rates vary with T and consistent with thermal and hydrotime models of biologicaltime, respiration rates per se were not the limiting factorin germination timing of tomato seeds. Key words: Lycopersicon esculentum Mill., tomato, germination, respiration, temperature, water potential     

Water Relations of Sitka Spruce Seedlings After Root Damage

Sitka spruce[Picea sitchensis(Bong.) Carr] seedlings were subjectedto varying degrees of root damage in a growth room, rangingfrom careful transplanting to exposure of the root system toair for up to 3 h. After replanting, transpiration (E), leafwater potential (₁) and growth of the shoot and root were measuredand observations made on plant survival. Some plants in the root exposure treatments died 20–85days after planting. In plants which eventually died, E wasdepressed directly after treatment, but ₁ showed a variableresponse. In some plants ₁ decreased from —8·0x 10⁵ to —30 x 10⁵ Pa after only 10 days but in othersthere was little change in ₁ for 50 days. In spite of the maintenanceof a high water potential in some of the latter plants for longperiods, no root or shoot growth occurred. In plants which lived, the root damage reduced root and shootgrowth relative to untreated controls, and most treatments stronglydepressed E but had little or no effect on ₁. The changes of E and ₁ in treated plants suggest that the suppressionof E was often independent of ₁ although water stress eventuallydeveloped in some of the severely treated plants. Sitka spruce, Picea sitchensis (Bong.)Carr, water relations, root damage, transpiration, leaf water potential     

Water Relations of Potato Leaves. II. Pressure--Volume Analysis and Inferences About the Constancy of the Apoplastic Fraction

Water relations characteristics of potato (Solanum tuberosumL. cv. Bintje) leaves were determined from pressure—volumeanalysis using a pressure chamber. Turgor was 077 MPa and thebulk volumetric modulus of elasticity 81 MPa at full turgidity;turgor loss occurred when water potential () had declined to–087 MPa at a relative water content (RWC) of 0912;the apoplastic water fraction (A) was 0235. As is usually found,there was a linear relation between 1/ and RWC beyond turgorloss. This finding supports the assumptions of the constancyof A during leaf dehydration. Beyond turgor loss the difference between and [measured afterfreezing and thawing (_d)] was about 01 MPa. This differencedid not increase as the leaf water content decreased. This resultcontradicts the constancy of A. It was concluded from calculations with a simple model of leafdehydration that analysis of the relation between and _d providesmore insight in the changes in the apoplastic fraction thanthe relation between 1/ and RWC. Research on the size of theapoplastic fraction and its changes with water potential wouldcomplement current understanding of leaf water relations. Solanum tuberosum, L., water potential, pressure chamber, osmotic potential, pressure potential, relative water content, apoplast, symplast     

Molecular dynamics simulations of oligosaccharides and their conformation in the crystal structure of lectin-carbohydrate complex: importance of the torsion angle {psi} for the orientation of {alpha}1,6-arm

The conformation of the heptasaccharide Man-1,6-(Man-1,3)(Xyl-ß1,2)-Man-ß,4-GlcNAc₂-ß1,4-(L-Fuc-1,3)-GlcNAc₁,the carbohydrate moiety of Erythrina corallodendron lectin (EcorL),the hexasaccharide Man-1,6-(Man-1,3) (GlcNAc-ß1,4)-Man-ß1,4-GlcNAc-ß1,4-GlcNAcand their disaccharide fragments have been studied by moleculardynamics (MD) simulations for 1000 ps with different initialconformations. In the isolated heptasaccharide, the most frequentlyaccessed conformation during MD has a value of 180° aroundMan-1,6-Man linkage. This conformation is stabilized by theformation of a hydrogen bond between the carbonyl oxygen ofGlcNAc₂ with the O3/O4 hydroxyls of the 1,6-linked mannose residue.The conformation of the heptasaccharide found in the crystalstructure of the EcorL-lactose complex (Shaanan et al., Science,254, 862, 1991), that has a value of 76° around Man-1,6-Manlinkage, is accessed, although less frequently, during MD ofthe isolated oligosaccharide. The ,, = 58°,–134°,–60°conformation around Man-₁,6-Man fragment observed in the crystalstructure of the Lathyrus ochnrs lectin complexed with a biantennaryoctasaccharide (Table I in Homans,S.W., Glycobiology, 3, 551,1993) has also been accessed in the present MD simulations.These values for the 1,6-linkage, which are observed in theprotein-carbohydrate crystal structures and are accessed inthe MD simulations, though occasionally, have not been predictedfrom NMR studies. Furthermore, these different values of leadto significantly different orientations of the 1,6-arm for thesame value of . This contrasts with the earlier predictionsthat only different values of can bring about significant changesin the orientation of the ₁,6-arm. The MD simulations also showthat the effects of bisecting GlcNAc or ß1,2-xyloseare very similar on the ₁,3-arm and slightly different on the₁,6-arm. bisecting GlcNAc carbohydrates glycoprotein lectinsaccharide complex     

Application of N-terminally Truncated DNA Polymerase from Thermus thermophilus ({Delta}Tth Polymerase) to DNA Sequencing and Polymerase Chain Reactions: Comparative Study of {Delta}Tth and Wild-Type Tth Polymerases

N-Terminally truncated DNA polymerase from Thermus thermophilus(Tth polymerase) lacking 5'-3' exonuclease activity was usedfor DNA sequencing and polymerase chain reaction (PCR). In contrastto the high background of the sequencing ladder observed withthe wild-type Tth polymerase, Tth polymerase gave readable sequencingpatterns which extend up to more than 500 bases from the primersite on cycle sequencing and automated sequencing. The Tth polymerasewas used for the standard and mutagenic PCR, and net amplificationof the DNA and the mutations accumulated during PCR were analyzed.Under mutagenic PCR, the mutation rates were 7.0 x 10^–4(Tth) and 8.3 x 10^–4 (Tth) per nucleotide per cycle ofamplification, which were 4–9 times higher than the ratesunder standard PCR.     

Wilting of Leaves in Vicia faba L.

Wilting of leaves of Vicia faba L., which occurs when the pressurepotential (_p) is zero, and the leaf-water potential () at wiltingboth depend entirely upon the solute potential at incipientplasmolysis (_so) and not on soil-water status. Wilting in V.faba is acropetal; this is consistent with the hypothesis thatthere is a gradient of decreasing _so up the plant and that wateris transferred from the lower to the upper leaves, hasteningthe overall water loss from the lower leaves to the point when_p is zero. The gradient in _so up the plant is of the order of3–8 bar. It is proposed that wilting when _p>0 (i.e. > _so) shouldbe ‘apparent wilting’ and that when _p0 (i.e. _so),‘true wilting’.     

Changes with Age in the Status of Water and Tissue Characteristics in Individual Leaves of Vicia faba L.

Leaf-water potential () and relative water content (R) of leavesat different position on the plant were different on any oneday. This was because the effect of environment on and R andof age on physical and chemical properties of tissues variedwith the position of the leaf. Changes with age in the physicaland chemical properties of a leaf depended on the time it emergedin the life of the plant.     

The Derivation of the Cell Wall Elasticity Function from the Cell Turgor Potential

An equation is derived expressing average turgor pressure ofa leaf (_p) as a function of relative water content (RWC). Basedon this derivation, the relationships of the bulk elastic modulus(_v) and both RWC and _p, are formulated and discussed. The bulkelastic modulus (_v) becomes zero for _p = 0, that is at the turgorloss point for the leaf. At full water saturation the valueof ev is proportional to the water saturation turgor potential_p(max). The factor relating _P and _v (structure coefficient ,Burstrom, Uhrstr?m and Olausson, 1970) changes only very littlefor values of _p, which are not too close to zero. An exampleis given for the calculation from experimental data of the turgorpressure function, the structure coefficient function, and the_v function. Key words: Cell wall, Turgor pressure, Bulk elastic modulus