Characterization of RNA-dependent RNA Polymerases in Healthy and Tobacco Mosaic Virus-infected Tomato Plants

Healthy tomato plants were shown to contain high levels of RNA-dependentRNA polymerase activity, mainly in a ‘soluble’ form,but also partly in a ‘ bound’ form. The ‘bound’enzyme was solublized by EDTA treatment. Both forms of enzymewere partially purified and characterized. The ion and pH optimaof the two forms were identical at all stages of purification.Both enzymes exhibited uridylyl transferase activity, whichmade up 35 per cent of total incorporation. Infection with tobacco mosaic virus (TMV) increased activityof ‘soluble’ enzyme by twofold, and of solubilized‘bound’ enzyme by less than twofold. Uridylyl transferaseactivity was also increased by infection. General propertiesof the enzymes were unaltered by infection with one exception:in the presence of TMV RNA as added template, the ‘soluble’enzyme from infected plants incorporated ³H-UTP into productswith the electrophoretic properties and RNase sensitivitiesexpected for replicative form and replicative intermediate ofTMV. ‘Soluble’ enzyme from healthy plants, and solublized‘ bound’ enzyme from either healthy or infectedplants did not synthesize these products. The ‘soluble’ and solubilized ‘bound’enzymes behaved differently on ion-exchange chromatography.Under the conditions used, ‘soluble’ enzyme didnot bind to the column, whereas solublized ‘bound’enzyme did. No differences in chromatographic behaviour werefound between enzymes from healthy or infected plants. Withboth ‘soluble’ and solublized ‘bound’enzymes, the uridylyl transferase activity co-chromatographedwith the polymerase activity. Tomato, Lycopersicon esculentum, RNA-dependent RNA polymerase, tobacco mosaic virus, tobacco mosaic virus replicase     

RNA secondary structures: comparison and determination of frequently recurring substructures by consensus

A method for assessing the preserved stem - loops of RNA secondarystructures is presented. Frequently recurring helical stemsin a set of secondary structures resulting from the simulatedfolding process of a given RNA are assessed and consensus structuralmotifs can then be selected to construct a secondary structureof the RNA. Alternatively, it can be applied to a series of‘optimal’ and ‘suboptimal’ secondarystructures computed using the dynamic program developed by Williamsand Tinoco. To demonstrate the power and the usefulness of theprogram we give examples of this procedure. Received on October 28, 1987; accepted on April 2, 1989     

Plant Morphology: The Historic Concepts of Wilhelm Troll, Walter Zimmermann and Agnes Arber

Recent molecular systematic and developmental genetic findingshave drawn attention to plant morphology as a discipline dealingwith the phenotypic appearance of plant forms. However, sincedifferent terms and conceptual frameworks have evolved overa period of more than 200 years, it is reasonable to surveythe history of plant morphology; this is the first of two paperswith this aim. The present paper deals with the historic conceptsof Troll, Zimmermann and Arber, which are based on Goethe'smorphology. Included are contrasting views of ‘unity anddiversity’, ‘position and process’, and ‘morphologyand phylogeny’, which, in part, are basic views of currentplant morphology, phylogenetic systematics and developmentalgenetics. Wilhelm Troll established the ‘type concept’and the ‘principle of variable proportions’. Hehas provided the most comprehensive overview of the positionalrelations of plant forms. Agnes Arber started from the universaldynamics of life and attempted to describe all structures asprocesses. She paid attention to ‘repetitive branching’,‘differential growth’, and ‘parallelism’.As a result she has recently been rediscovered by developmentalbotanists. Walter Zimmermann rejected any metaphysical influenceon plant form and instead called for objective procedures. Hewas mainly interested in phylogenetic ‘character transformation’and the ‘reconstruction of genealogical lines’.Guided by the example of flower-like inflorescences, a futurepaper will deal with functional and developmental constraintsinfluencing plant forms. Recent morphological concepts (‘trialectical’,‘continuum’/‘fuzzy’, ‘processmorphology’) will be discussed and related to currentmorphological and developmental genetic research. Copyright2001 Annals of Botany Company Plant form, plant morphology, natural philosophy, homology, phylogeny, Goethe, Troll, Arber, Zimmermann, typology, character transformation, differential growth, complementarity     

A toolkit for creating three-dimensional RNA pencil models

The advantages of nucleotide-resolution models over atomic-resolutionand cylinder models of large RNA structure are discussed, anda toolkit of RNA substructures for use in building ‘pencil’models is described. Prefabricated elements from the toolkitcan be used to quickly assemble complex RNA structure modelsfor the visualization of known RNA structures, or for exploringpotential tertiary structure configurations based on secondarystructure and other information.     

Effect of large- and of small-bodied zooplankton on phytoplankton in a eutrophic oxbow

Macrozooplankton and microzooplankton effects on the phytoplanktonwere measured in situ in a eutrophic lake. Indigenous phytoplanktonwere incubated for 5 days in 301 mesocosms with either the macro-and microzooplankton (complete), microzooplankton only (micro)or no zooplankton (none). Changes in phytoplankton biovolumewere investigated. Rotifer densities became significantly higherin the ‘micro’ treatment than in the ‘complete’and ‘none’ treatments. Total algal biovolume changedlittle in the ‘complete’ and ‘none’treatments, but increased significantly in the ‘micro’treatment. The results suggest that macrozooplankton (Daphniamagna) suppressed it and microzooplankton (Keratella cochlearis)enhanced it. They had opposite net effects on the phytoplankton.Suppression of microzooplankton by Daphnia probably had an indirectnegative effect on the phytoplankton.     

The Extraction, Solubility, and Characterization of Two Groups of Barley Storage Polypeptides

This paper reports further studies on the characteristics ofthe storage protein fraction (hordein) of barley. Hordein consistsof two groups of polypeptides (termed ‘B’ and ‘C’)coded by two separate but linked loci. Whereas the ‘C’polypeptides are readily soluble and extracted in 60% (v/v)ethanol at room temperature, the ‘B’ group is moresoluble in, and therefore more efficiently extracted by, 50%(v/v) propan-1-ol or 45% (v/v) propan-2-ol at elevated temperaturesand in the presence of 2-mercaptoethanol. However, the mostefficient conditions for hordein extraction (50% propan-1-ol+ 2% (v/v) 2-mercaptoethanol at 60 °C) also extract somecontaminating non-hordein polypeptides resulting in an apparentlyincreased lysine content of the hordein fraction. Amino acid analysis of the purified ‘B’ and ‘C’hordein groups shows that, whereas ‘C’ hordein containsmore glutamate + glutamine, proline, and phenylalanine than‘B’ hordein, it contains only traces of lysine andsulphur amino acids in contrast to ‘B’ hordein whichcontains 0·5% lysine 0·6% methionine, and 2·5%cysteine. Equilibrium sedimentation analyses carried out on the purified‘B’ and ‘C’ groups indicates that thepreparations were reasonably monodisperse with molecular weightsof approximately 32 000 and 52 000 respectively. These valuesare considerably lower than those previously determined by SDS-PAGE.     

A program for predicting significant RNA secondary structures

We describe a program for the analysis of RNA secondary structure.There are two new features in this program. (i) To get vectorspeeds on a vector pipeline machine (such as Cray X-MP/24) wehave vectorized the secondary structure dynamic algorithm. (ii)The statistical significance of a locally ‘optimal’secondary structure is assessed by a Monte Carlo method. Theresults can be depicted graphically including profiles of thestability of local secondary structures and the distributionof the potentially significant secondary structures in the RNAmolecules. Interesting regions where both the potentially significantsecondary structures and ‘open’ structures (single-strandedcoils) occur can be identified by the plots mentioned above.Furthermore, the speed of the vectorized code allows repeatedMonte Carlo simulations with different overlapping window sizes.Thus, the optimal size of the significant secondary structureoccurring in the interesting region can be assessed by repeatingthe Monte Carlo simulation. The power of the program is demonstratedin the analysis of local secondary structures of human T-celllymphotrophic virus type III (HIV). Received on August 17, 1987; accepted on January 5, 1988     

CORRECTIONS, VOLUME 29

Part 1, under the frontispiece portrait of Dr. N. B. Eales,the words ‘President 1948–1951’ should havebeen added. Page 103, line 49, for ‘Newton Collection’ read‘Norman Collection (Canon Norman)’. 185, line 37, for ‘capillaris’ read ‘capillacca’. 188, Table 1, for ‘bemoralis’. read ‘nemoralis’. 188, Table 2, for ‘Cochlicella acuta (Müll)? ventrosa(Fér.)’ read ‘Cochlicella ventrosa (Fér.)’. 191, line 24, for ‘araheo-’ read ‘archeo-’.     

Uptake and Metabolism of Sugars by Suspension cultured Catharanthus roseus Cells

p. 186, right column line 11 ‘27.2 KBq’ change to‘37.2 MBq’ p. 187, left column line 8 ‘27.2 KBq’ change to‘37.2 MBq’ line 10 ‘13.6 KBq’ change to ‘18.6 MBq’ line 11 ‘13.6 KBq’ change to ‘18.6 MBq’ line 21 ‘89%’ change to ‘80%’ right column line 24 ‘CLC-NH₄’ change to ‘CLC-NH₂’ P. 189, Table 1 appeared incorrectly: it should appear as indicated.     

Theodoropoulos, D.I. Invasion biology. Critique of a pseudoscience.

The back cover of this book states that ‘contrary to theclaims of the nativists, research shows that man-dispersed speciesincrease biological diversity, benefit ecosystems, and act asan important force for healing the planet’. This is anuncompromising statement, and David Theodoropoulos divides hisdevelopment of the arguments supporting this statement intothree parts. Part I (Chapters 1–6) is ‘Nature, Dispersaland Reaction’. Part II (Chapters 7 and 8) is ‘Why?Psychology, Politics and Pseudoscience’. Part III (Chapters9–11) is ‘Humanity and Diversity’. There isalso an ‘Introduction’ including a summary of findingsand ‘An outline for a new theory of anthropogenic dispersal’,     

Confusion in the use of the taste adjectives 'sour' and 'bitter'

Subjects from Britain and America were tested on their skillat applying the taste adjectives ‘sweet’, ‘sour’,‘salty’, and ‘bitter’ to clearly distinguishablesolutions of sucrose, citric acid, NaCl and quinine sulphate,respectively. The main error that occurred was calling citricacid ‘bitter’ while the tendency to call quininesulphate ‘sour’ was not so common; this is the wellknown sour-bitter confusion. A sour-salty confusion was alsonoted as well as a tendency to call citric acid ‘sweet’.All these confusions were rectified by mere definition usingstandards. Skill at applying taste adjectives was not alwaysfound to be consistent over time. More errors occurred at lowersolution concentrations, even though stimuli were clearly distinguishable;indistinguishability of stimuli may account for some confusionsin other studies. ^*Present address: Department of Psychology, University of Bristol,Bristol BS8 1HH, England.     

Cultivar Differences in the Performance of Bean Seedlings at Suboptimal Temperatures

Growth analysis of plants raised under controlled environments(10–5, 12, 15, 18 and 20 °C, and 21 h photoperiod)was used to examine whether varietal differences in the minimumgermination temperature of four bean cultivars persist duringgrowth at suboptimal temperatures. A method to estimate theminimum vegetative growth temperature, based on axis relativegrowth rate, was developed. In order to compensate for ontogeneticdrift, the harvests were conducted at the same stage of developmentof the plants. Axis relative growth rates, reduction rates ofthe cotyledons and other growth parameters were calculated inorder to compare the cultivars. Cultivar ‘Marschall’showed better growth potential at 12 °C than the others,‘Pergousa‘ at 15 °C, and ‘Marschall’,‘Olsok’ and ‘Pergousa’ at 18 and 20°C. The effect of temperature on axis RGR was similar for‘Marschall’, ‘Olsok’ and ‘Pergousa’(Q₁₀ = 2·1) and more pronounced than for ‘Processor’(Q₁₀ = 1·3). Although there were significant differencesin the growth parameters among the cultivars within each temperatureused, the differences did not correspond with the differencesduring germination at low temperatures. The minimum vegetativegrowth temperature was close to 10 °C for all the cultivarstested. Phaseolus vulgaris L., beans, suboptimum temperature, growth analysis, minimum germination temperature, minimum vegetative growth temperature     

Effects of Inorganic Phosphate on the Utilization of Sucrose by Suspension-cultured Catharanthus roseus Cells

The metabolic fate of [U-¹⁴C]sucrose in suspension culturesof Catharanthus roseus cells was monitored for 96 h after thecells were transferred to fresh complete (‘+Pi’)or to phosphate-deficient Murashige and Skoog (‘–Pi’)medium. Sucrose was hydrolysed extracellularly to glucose andfructose. The rate of uptake of sugars by the cells was 1.5–3times higher in ‘+Pi’ culture than in ‘–Pi’culture. Little difference in the rate of incorporation of radioactivityinto the ethanol-soluble fraction was found between the ‘+Pi’and ‘– Pi’ cultures during the initial 24h of culture, but after 48 h the rate in ‘ +Pi’cultures was higher than that in ‘–Pi’ cultures.Incorporation of radioactivity into ethanol-insoluble macromoleculeswas always significantly higher in the cells in ‘+Pi’cultures than in those in ‘–Pi’ cultures.The results suggest that Pi strongly affects the utilizationof sugars by cultured plant cells through the stimulation oftransport of sugars as well as through the activation of metabolism. Catharanthus roseus, Madagascar periwinkle, suspension culture, inorganic phosphate, sucrose, transport, metabolism     

The Nitrogen Content of Plants and the Self-thinning Rule of Plant Ecology: A Test of the Core-skin Hypothesis

The ‘core-skin’ hypothesis postulates that secondarilythickened plants behave energetically as an inert ‘core’covered by an active ‘skin’, the ‘skin’being two-imensional, the ‘core’ three-dimensional.This would explain the ‘self-thinning ‘or‘–3/2’ rule of plant ecology, that is, the tendencyfor log (dry weight per plant) and log (number of plants perunit area) to progress along a straight line relationship, withslope = – 3/2’. The hypothesis was tested as follows. Plant nitrogen contentwas used as an estimate of the mass of ‘skin’ perplant, and dry weight as an estimate of the mass of the ‘core’.As plants mature the slope of the relationship between y = log(mass of nitrogen per plant) and x = log (mass of dry matterper plant) is expected to decline from an initial value of 1.0towards a final value of 0.66. The intercept of the relationshipis expected to reflect the intrinsic content of ‘skin’per unit of ‘core’. Genotypic variation in thisparameter should cause genotypic differences in the maximumattainable yield of biomass per unit area. The expectations were investigated by fitting the function y= p+qx+r exp – x to 30 sets of data on plant nitrogencontent, plant weight and time in 18 different vegetables. Simplelinear regressions of y on x were fitted to more limited setsof data on weights and nitrogen contents of mature trees. Theexpectations were, with some minor exceptions, confirmed. Nitrogen, yield, plant competition, self-thinning     

Diurnal Phototropism in Leaves of Lavatera cretica L. under Conditions of Simulated Solar-Tracking

Diurnal laminar reorientation was followed in solar-trackingleaves of Lavatera cretica L. under simulated conditions. Asimulated ‘sun’ was moved over the lamina in a 180?arc in the vertical plane of the mid-vein, at an angular velocityof 15? h^–1 in a regime of 12-h photoperiods. In one groupof plants the petioles of the experimental leaves were arrangedto face ‘sunrise’, while in the other they werearranged to face ‘sunset’. At ‘sunrise’,the laminae in both groups, which were inclined towards theanticipated direction of ‘sunrise’, changed theirelevation towards the rising ‘sun’, resulting inprogressive reduction in the angle of incidence (AI) of lighton the laminar surface (AI= differential between laminar and‘solar’ elevation). As a result, laminar and ‘solar’elevation converged, and laminar reorientation gradually ceased,until the ‘solar’ elevation had passed the normalto the laminar surface (AI=0?). laminar reorientation was thenresumed, but its direction was reversed to follow the directionof ‘solar’ reorientation. During most of the remaining‘day’, laminar elevation (LE) trailed that of the‘sun’ by an average of 11?-14?. Laminar reorientationthen anticipated ‘sunset’ by starting to slow down60 to 90min in advance. During the 12-h dark period, the laminareoriented towards the anticipated direction of the subsequent‘sunrise’. The time-course of nocturnal reorientationwas qualitatively different in the two groups of experimentalplants. The time-course of diurnal phototropism under naturaland simulated conditions is analysed and compared and differencesand similarities between them are discussed. Key words: Diurnal phototropism, solar-tracking, vectorial excitation     

Relative Sensitivity and Tolerance of some Gladiolus Cultivars to Sulphur Dioxide

Five Gladiolus cultivars, namely ‘Aldebaran’, ‘BrightEye’, ‘Illusion’, ‘Manisha’ and‘Manmohan’, were exposed to 1 and 2 µg l^–1sulphur dioxide to test their relative-sensitivity toleranceto the pollutant Plants were fumigated experimentally for 2h daily Foliar injury symptoms were observed first in ‘Manisha’followed by ‘Aldebaran’ and ‘Illusion’at the higher dose Photosynthetic pigments and leaf extractpH were significantly decreased, particularly in ‘Manisha’and ‘Illusion’ Overall disturbances in the plantmetabolism due to SO₂ treatment led to retarded growth of plants,as evident from decreased shoot length and phytomass valuesThe order of sensitivity of the five Gladiolus cultivars toSO₂ was as follows, with the greatest first Manisha, Illusion,Aldebaran, Bright Eye, Manmohan Cultivars, Gladiolus, sensitivity, sulphur dioxide, tolerance     

The Influence of the Plant Growth-regulator, 2-Methyl-4-Chlorophenoxyacetic Acid, on the Metabolism of Carbohydrate, Nitrogen and Minerals in Solanum lycopersicum (Tomato)

The effects of the foliar application of phytocidal concentrationsof 2-methyl-4-chlorophenoxyacetic acid (MCPA) on change in totaldry weight, and in ‘available carbohydrate’ (starch,‘total’ and ‘reducing’ sugars), totalnitrogen, phosphorus, potassium, calcium, and magnesium of ‘tops’and roots of tomato plants have been followed over a periodof 14 days following spraying. There were two main treatments—‘nutrient’(nutrient supply to roots continued after spraying) and ‘water’(distilled water only supplied to roots after spraying) and‘water’ (distilled water only supplied to rootsafter spraying)—the sub-treatments consisting of ‘MCPA’versus ‘no-MCPA’ for each of the main treatments.Twelve different times of sampling were used. In analysing the present data, the quantity ‘residualdry weight’ (total dry weight less ‘available carbohydrate’),which was originally introduced by Mason and Maskell as a basisof reference for analyses of plant organs in short-period experimentsnot involving appreciable growth, has been used as an estimateof the permanent structure of plant growth. This new use ofthe ‘residual dry weight’ basis has brought outimportant features which were obscured when the data were leftin their primary form (as percentages of total dry weight oramounts per plant). Growth, as measured by increase in ‘residual dry weight’,was greatly inhibited by 2-methyl-4-chlorophenoxyacetic acidshortly after spraying, in both the presence and the absenceof nutrient. In the presence of 2-methyl-4-chlorophenoxyacetic acid, netassimilation rate (estimated as rate of increase in total dryweight per gram ‘residual dry weight’ of the ‘tops’)was greatly diminished while uptake of total nitrogen and ofP₂O₅ (estimated as increase in total nitrogen or of P₂O₅ ofthe whole plant per day per 1 g. ‘residual dry weight’of the roots) appeared to undergo a similar but much smallerdiminution. It seemed probable, however, that in the presenceof MCPA a larger proportion of the carbohydrate actually formedwas utilized for synthesis of aminoacids and protein. In the plant as a whole there was no evidence of actual depletionof ‘available carbohydrate’ as a result of MCPAtreatment, this fraction showing a steady increase in all treatmentsthroughout the experiment. The rate of increase was, however,much reduced by MCPA treatment. The ‘tops’ presentedmuch the same picture as the whole plant, but for the rootsthe situation was quite different. While the roots of the ‘no-MCPA’plants and also of the ‘MCPA-water’ plants showeda steady increase in available carbohydrate, those of the ‘MCPA-nutrient’plants rose only very slightly (from the initial value of 8mg. per plant to about 10 mg.) during the first 2 days, andthen in the next 2 days declined to a value (about 6 mg.) belowthe initial and remained at this low level for the rest of theexperiment. It is suggested that the phytocidal effect of 2-methyl-4-chlorophenoxyaceticacid in the presence of nutrient may be due to depletion ofthe ‘available carbohydrate’ supplies in the roots,which is shown to be brought about, in part, by reduced transportfrom the tops, and partly by the relatively greater utilizationof the carbohydrate present. These results offer an explanationfor the facts that plants showing vigorous growth are more easilykilled by MCPA and that perennial plants, particularly thosewith storage tissues in their roots, are more resistant. Further,they suggest the useful practical application that MCPA treatmentshould be given when the carbohydrate reserves of the rootsare at a minimum. For perennial plants, conditions might beexpected to be optimal for the application of MCPA in late spring,at a time when the first ‘flush’ of growth is slowingdown and before any appreciable new reserves of carbohydratehave been accumulated. It was also shown that 2-methyl-4-chlorophenoxyacetic acid preventedthe net synthesis of starch, but still permitted an appreciablenet formation of sucrose. 2-methyl-4-chlorophenoxyacetic acid appeared to have no effecton the uptake of potassium, calcium, or of magnesium. The lackof effect on potassium is contrasted with the previous observationby Rhodes, Templeman, and Thruston (1950) that sub-lethal concentrationsof MCPA, applied over a relatively long period to the rootsof tomato plants, specifically depressed the uptake of potassium.     

Properties of Mitochondria Prepared by Two Different Isolation Procedures From the Cotyledons of Germinating Bean Seedlings

Mitochondria from 1-day-old bean cotyledons were isolated bya ‘slow’ isolation procedure involving a wash andby a ‘rapid’ procedure. The mitochondria isolatedby the ‘rapid’ procedure were more tightly coupledthan those isolated by the ‘slow’ procedure. Anexogenous supply of cytochrome c or NAD was shown to improvethe activity of mitochondria isolated by the ‘slow’procedure, but not those isolated by the ‘rapid’procedure. The phosphorylative abilities of the latter weremuch greater than the former and were retained for longer periods.It was found that there was a leakage of NAD out of the mitochondria.     

Restriction map construction using a 'complete sentences compatibility' algorithm

We have developed a new algorithm ‘Complete sentencescompatibility’ (CSC) which uses single and double digestionfragments to rapidly determine restriction maps of circularDNA. From possible combinations of fragments of each simpledigestion, which we call ‘sentences of decomposition’,we construct a restriction map which combines the sentenceswhile taking into account compatibility rules. The algorithmcan also deal with experimental errors of fragment weight andcan suggest solutions that account for non-readable bands (fragmentsof zero length or multiple bands) on the gel. Because experimentsusing pairs of restrictive enzymes often result in multiplesolutions, a complementary algorithm tries to reduce the numberof proposed solutions by establishing consensus maps. The restrictionmap construction algorithm was tested on real cases, some containingmore than fifteen fragments. Execution times range from 1 –10 s on an IBM PC compatible microcomputer. Received on July 21, 1987; accepted on December 31, 1987