A combined morphological and molecular approach to Nitzschia varelae sp. nov., with discussion of symmetry in Bacillariaceae

A previously unknown member of the Bacillariaceae was discovered almost simultaneously in four different brackish coastal wetlands on the Atlantic and Mediterranean coasts of the Iberian Peninsula. It appears to tolerate a wide range of salinities but was never common in samples where it occurred. The frustules were consistently hantzschioid (i.e. with the raphe systems always on the same side of the frustule) and the valve outline was asymmetrical about the apical plane, two features that have until recently been considered characteristic of Hantzschia. Molecular phylogenies based on rbcL and LSU rDNA indicated, however, that the new species does not belong in Hantzschia but among the several disparate lineages that comprise the paraphyletic genus Nitzschia. This finding, coupled with the recent discovery of other diatoms with constant hantzschioid symmetry but with a morphology very similar to the type species of Nitzschia, is discussed in relation to the status and characterization of Hantzschia as an independent genus. It is concluded that, while a core of hantzschioid species may exist that can be classified together, corresponding to the traditional understanding of the genus Hantzschia, there is no single morphological feature common to all of them that can be used to diagnose the group and differentiate it from the various hantzschioid lineages that are separate from true Hantzschia and currently placed in e.g. Nitzschia or Cymbellonitzschia. Testing whether a hantzschioid species does or does not belong to Hantzschia will in many cases require molecular evidence. Although the new coastal species does not belong to the same lineage as the type species of Nitzschia, N. sigmoidea, it is described for the moment as N. varelae Carballeira, D.G. Mann & Trobajo, sp. nov., until there is a better understanding of generic limits in the Bacillariaceae following a wider molecular and morphological survey of that family.     

Studies on Mature Seeds of Cuscuta pedicellata and C. campestris by Electron Microscopy

The seeds of Cuscuta pedicellata have been investigated by transmissionand scanning electron microscopy. Additional observations havebeen made on seeds of C. campestris by SEM only. The seed coatconsists of an outer single epidermis, two different palisadelayers, and an inner multiparenchyma layer. The outer epidermalwall in C. pedicellata has a thick cuticle and zones rich inpectic substances. The thicker ‘U-shaped’ cell wallsin the outer palisade layer are strengthened by a wall layerof hemicellulose. The inner palisade layer has thick walledcells with a ‘light line’. The inner cell wall ofthe compressed multiparenchyma layer has a thin cuticle. A fairlythick cuticle is positioned directly on the endosperm surface.The aleurone cell walls are different from the remaining endospermwalls. The latter are thick and believed to be of galactomannans.There is a ‘clear’ zone between the plasmalemmaand the cell wall in the aleurone cells. The embryo cells arepacked with lipids and proteins. In Cuscuta campestris mostendosperm has been absorbed during the seed development. Theembryo apex has two minute leaf primordia. The features of theCuscuta seeds are discussed in relation to functional and environmentalconditions. Cuscuta pedicellata, Cuscuta campestris, seed, seed coat, cuticle, cell walls, endosperm, aleurone cells, galactomannan, embryo, TEM, SEM     

A Rapid Method for the Determination of the Leaf Area of Oxalis corniculata L. and Tephrosia purpurea (Linn.) pers.

A rapid simple method has been described for the determinationof leaf area of Oxalis corniculata L. (Oxalidaceae) and Tephrosiapurpurea (Linn.) pers. (Leguminoseae) by evaluating area = a²x0.866and a²x0.335 respectively for the two plants, where a representsa single linear measurement. The relation has been derived afterintegrating the equation of the curve r² = a² cos 2 coveringthe area of each leaflet. Areas of a large number of leaflets have been determined withthe help of two methods (1) Planimeter method, (2) Evaluationmethod. The significance of the difference between the two valuesfor a particular leaf has been statistically examined and itappears that the former method can be replaced by the latterin the case of these plants.     

Action Potentials in Lupinus angustifolius L. Shoots: II. DETERMINATION OF THE STRENGTH-DURATION RELATION AND THE ALL-OR-NOTHING LAW

In Lupinus shoots an electrical stimulus (d.c.) produces a potentialwave analogous to the action potential wave (AP) in stimulatedsimple plant cells or in nerves. The method used (Paszewskiand Zawadzki, 1973a, b) is similar to those applied in neurophysiologyin research on the excitability of nerves. In the present paper the strength-duration relation (Eqn. (1))of the excitation in Lupinusitalic has been used to calculatethe values of the rheobase, chronaxie, and useful time of pulse.The rheobase, as a threshold value of the stimulus, is examinedin terms of the applicability of the all-or-nothing law. The results suggest that the origin and propagation of AP inplants and in nerves may occur in a similar way. The analogybetween the impulse propagation in nerve trunks and in Lupinusshoots is discussed.     

Temporal and Spatial Heterogeneity in the Accumulation of Anthocyanins in Cell Cultures of Catharanthus roseus (L.) G.Don.

The accumulation of anthocyanins, a group of pigmented secondarymetabolites, in cell cultures of the Madagascar periwinkle Catharanthusroseus has been investigated. In these cultures it was foundthat anthocyanin accumulation was restricted to the post-divisionphase of the culture growth cycle, during which the culturesbecame deep purple in colour. As a result of anthocyanin visibilityit has been possible to ascertain that accumulation of thesemetabolites occurred in only a small proportion of the cellpopulation. Approximately 10% of cells regularly accumulateddetectable levels. Considerable variation within this ‘productive’population was observed and using a standard integrating microdensitometerit has been possible to quantify directly this heterogeneityand compare it with data obtained from whole plants. Analysishas revealed that the variation in both intracellular anthocyanincontent and concentration in cell cultures was much greaterthan that observed within tissues of mature plants. Significantdifferences in mean values were however found between the wholeplant tissues. The relevance of this temporal and spatial heterogeneityobserved in vitro to our understanding of the control of secondarymetabolite accumulation and to the potential use of tissue culturesystems as a means to produce these compounds is discussed. Key words: Heterogeneity, anthocyanins, cell culture     

Total Uptake and Incorporation into DNA of 3H-Thymidine, 3H-Deoxyuridine and 3H-Thymine in the Primary Root of Vicia faba L.: I. INTACT ROOTS

Total uptake and incorporation of ³H-thymidine, ³H-thymine,and ³H-deoxyuridine into DNA have been investigated in the apical3 cm of the primary root of Vicia faba. Evidence has been obtainedthat endogenous TdR in these roots may be transported eitherapically or basally; apical movement being greater than movementfrom the apex towards the base of the root. The results havebeen discussed with respect to the possible distribution ofendogenous pools of thymidine, thymine, and deoxyuridine inthe primary root of V. faba.     

Direct Somatic Embryoid Formation on Immature Embryos of Trifolium repens, T. pratense and Medicago sativa, and Rapid Clonal Propagation of T. repens

By direct somatic embryogenesis in vitro a clone of asepticplantlets can be raised from a single immature embryo of Trifoliumrepens (white clover) within about 6 weeks of pollination. Embryoidsare induced directly from intact zygotic embryonic tissue ona culture medium containing 0·025 or 0·05 mg 1^–1BAP and 1·0 g 1^–1 yeast extract. Similar directsomatic embryogenesis has also been achieved for Trifolium pratense(red clover) and Medicago sativa (lucerne). Applications ofembryo propagation by direct somatic embryogenesis are discussed,particularly in relation to multiple screening of host genotypesfor analysis of host/pathogen and legume/Rhizobium interactions. Trifolium repens L., Trifolium pratense L., Medicago sativa L., clover, lucerne, tissue culture, embryoid, somatic embryogenesis, legumes     

Changes in the Distribution of Plasmodesmata in Developing Fibre-tracheid Pit Membranes of Sorbus aucuparia L.

The distribution of plasmodesmata in cambial pit fields anddeveloping fibre-tracheid pit membranes in Sorbus aucupariaL. has been studied using conventional and high voltage transmissionelectron microscopy. In cambial pit fields, plasmodesmata arewidely separated while, during cell enlargement and the stageof secondary wall formation, they occur exclusively in a denselypacked cluster, eccentrically located on the pit membrane ina thickening of primary wall material. It is not clear at themoment whether the cluster arises by aggregation of the cambialpit field plasmodesmata, or by de novo formation in a localizedregion coupled with loss of plasmodesmata outside this region.The significance of these changes is discussed. Sorbus aucuparia L., plasmodesmata, pit membranes, xylem differentiation     

STEROLS OF CHLORELLA. I. THE NATURALLY OCCURRING STEROLS OF CHLORELLA VULGARIS, C. ELLIPSOIDEA, AND C. SACCHAROPHILA

The characteristics of the sterols naturally occurring in threespecies of Chlorella were examined. The algae were grown heterotrophicallyon glucose. Sterols were extracted and isolated from the lipidfraction and were characterized by means of chemical and physicaltests. Chlorella vulgaris contained three sterols. Only the principalone, chondrillasterol, was identified. Chondrillasterol hasbeen isolated previously from the genus Scenedesmus. Chlorella ellipsoidea and Chlorella saccharophila were foundto contain sterols with ß-oriented alkyl groups atC-24 in contrast to the -oriented groups commonly found in higherplants. Poriferasterol was identified as the principal sterolof both algae. Clionasterol and 22-dihydrobrassicasterol wereidentified as the two secondary sterols present. None of thesesterols have previously been reported to occur in plants. Theisolation of 22-dihydrobrassicasterol has not been previouslyreported from any natural source. ¹Scientific Article A1153, Contribution No. 3623 of the Universityof Maryland Agricultural Experiment Station. ²This work has been supported in part by a grant from the NationalAeronautics and Space Administration.     

Further Observations on the Structure of Plant Cilia, by a Combination of Visual and Electron Microscopy

By combining visual microscopy of stained material with electronmicroscopy it has been possible to demonstrate fibrillar disintegrationin the cilia of Saprolegnia (front and back), Chlorosaccus (frontand back), Allomyces, and Olpidium. These represent three fungiand a yellow-green alga. All flagella showed eleven componentstrands, in most cases clearly differentiated into nine strandsand a central pair. In addition, stained and unstained materialof Saprolegnia and Chlorosaccus have been closely compared inthe electron microscope and the comparison has included observationson one and the same cell with die electron microscope and thelight microscope. The nature of the artifacts produced by staininghas been investigated and the value of the combination of thesevarious technical processes demonstrated. The phyletic importanceof the hairs on the front flagellum in the yellow-green algae,brown algae, and water moulds is discussed.     

Experimental and Analytical Studies of Pteridophytes: XXIII. The Induction of Buds in Ophioglossum vulgatum L.

A study has been made of the development of buds on isolatedroot and stem segments of Ophioglossum vulgatum. In both casesbuds may develop endo-genously. The anatomical details of developmentare fully described. The bearing of the facts on current theoriesof shoot initiation is discussed.     

Vegetable Plant Part Relationships. II. A Quantitative Hypothesis for Shoot/Storage Root Development

A simple quantitative formulation of the concept of the controlof partitioning of assimilated carbon by the behaviour of plantcomponents as competing sinks is developed. An equation, In s = + In r—t, relating shoot (s) and storage root (r) d. wts, and the lengthof growth period (t), is constructed by considering possiblefates of imported assimilates into different plant parts. Thevalues of the equations' parameters depend on the relative sinkactivities of the plant parts, tissue respiration rates andinitial weights of plant components. The equation closely fitteddata collected from a number of carrot and beet experimentsin which planting density had been varied. Estimates of shootand storage root maintenance respiration rates, derived fromthe parameter , were of the correct order of magnitude. Othersets of experimental data are also discussed in the light ofpredictions of the theory and possible uses and extensions ofthis approach to assimilate partitioning are briefly discussed. Daucus carota L., Beta vulgaris, carrot, red beet, partition of assimilated carbon, maintenance respiration, storage root     

Studies in the Diatom Genus Hantzschia 3. Infraspecific Variation in H. virgata

Infraspecific variation has been investigated in the marineepipelic diatom Hantzschia virgata using light and electronmicroscopy. Although the variation pattern in this species iscomplex and to some extent discontinuous, allowing the recognitionon phenetic grounds of four or five discrete infraspecific taxaincluding vars. virgata (= H. insolita), gracilis, leptocephala(= capitellata) and wittii (= intermedia), it is concluded thatto split the species would be imprudent. Variation has beenfound not only in characters such as valve outline and size,or stria density, but also in ‘qualitative’ characterssuch as raphe, fibula and girdle structure, characters whichhave often been assumed to be constant within species, generaor taxa of higher rank: previous generalizations about the usefulnessof certain characters in distinguishing taxa of a given rankare seen to have been incautious. Hantzschia virgata (Roper) Grun., diatom taxonomy, infraspecific variation     

Late Transplanting and the Yield of Phaseolus vulgaris L.

The prediction that very high seed yields of dry beans (Phaseolusvulgaris L.) would be produced by the delayed transplantingof large plants has been tested in a factorial experiment withfour dates of transplanting and eight plant populations. Therewere significant differences in yield between transplantingdates and between population densities, and there was a significantdate-density interaction. At low plant densities (up to about30 plants m^–2) the three transplanted treatments yieldedless than the hand-sown controls, and late transplanting yieldedless than early. At the highest density the situation was reversed;all three transplanted treatments out-yielded the controls andlate transplanting tended to out-yield plants transplanted early.The biggest yield was 340 g seed m^–2 from a transplantedcrop grown at 35 plants m^–2. The data on yield fitted a modified rectangular hyperbola ofthe form where y is yield per unit area, p is the number of plants perunit area, t is the number of days between sowing and transplanting,and B_o, n, m, and p are arbitrary parameters. This equationaccounted for 91 per cent of the variation in yield with t andp. It is suggested that late transplanting had adverse effects,due to transplanting ‘shock’ and which were mostmarked at low plant densities; and beneficial effects, ascribableto an effect on plant ‘plasticity’, which were mostmarked at high plant densities. Possible physiological mechanismsof these effects are discussed. Phaseolus vulgaris, yield, density, transplanting     

Production of 2n Gametes in Diploid Subspecies of Dactylis glomerata L. 2. Occurrence and Frequency of 2n Eggs

Whereas polyploidy has been found in more than half the Gramineae,the occurrence of 2n gametes, a potentially major mechanismof polyploid production, has been rarely studied. In the presentwork, the frequency of 2n egg production in nine diploid (n= 2x = 14) subspecies of the polyploid complex Dactylis glomeratahas been estimated by determining the ploidy level of the progenyin 98 2X-4X crosses. The 2n egg origin of the 4X offspring wasverified using enzyme markers. The frequency of 2n egg productionin each subspecies was estimated from the number of 4X offspring,the number of florets in the seed plant and the fertility ofthe subspecies. The ploidy level of the progeny was determinedby chromosome counts or nuclear DNA quantification using flowcytometry. Plants producing 2n eggs were detected in 47% ofthe fertile crosses, involving eight subspecies. Variabilitywas related to individuals and not to subspecies. Average 2negg frequency was 0·49% for all crosses, and 1·53%for crosses where 2n egg production was detected. Individualfrequencies did not exceed 3·5%, with the exception of26% in one plant. Triploid block was higher and fertility, frequencyof high 2n gamete producers and average 2n gamete frequencywere lower in the 2X-4X crosses than in the opposite 4X-2X crossesinvolving the same plants, previously studied. Diplogynous anddiplandrous gamete production were not correlated with eachother. However, the widespread occurrence of plants producingboth kinds of 2n gametes in almost all the studied subspecies,as well as the presence of several individuals able to producehigh rates of 2n eggs or 2n pollen, support the hypothesis thattetraploids can be formed anywhere and anytime by sexual polyploidizationin diploid populations of Dactylis glomerata L. 2n eggs, polyploidy, Dactylis glomerata L., flow cytometry, sexual polyploidization     

Karyological Anatomy of Fibre Development in the Leaves of Agave americana L. var. marginata alba Trel.

From a study of 100 metaphase plates in root-tip squashes and75 fibre-cells in leaf-base squashes, it is found that the chromosomenumber, although having an irregular type of variation, commonlytend to occur as multiples of 15, the haploid number. In developingfibre-cells the tendency of complete endomitosis (resultingin 2n, 4n, and 8n numbers) is stronger than the partial endomitosis(resulting in 5n, 6n, 7n, etc.). Nuclear volumes of fibre-cellsalso roughly correspond to 2K, 4K, and 8K. Significant linearcorrelations between chromosome number and nuclear volume andbetween nuclear volume and fibre-cell volume has been obtained.Thus the development and differentiation of fibres involve completeand partial endomitoses, accompanied by somatic non-disjunctionphenomena.     

Studies on Foliar Penetration: IX. PATTERNS OF PENETRATION OF 2,4-DICHLOROPHENOXYACETIC ACID INTO THE LEAVES OF DIFFERENT SPECIES

The comparative patterns of penetration of 2,4-dichlorophenoxyaceticacid (2,4-D) into the leaves of Phaseolus vulgaris, Zea mays,Pisum sativum, Beta wlgaris, Helianthus annuus and Gossypiumhirsuium have been examined. Save for Zea and Gossypium where there is little change withthe stage of leaf development the rates of penetration intoboth surfaces decrease as the leaf matures. The relative ratesare dependent on the species and the age of the leaf but thereare differences between the surfaces. In Phaseolus the characteristicsof primary leaves differ from those of trifoliate leaves sinceonly in immature trifoliate leaves is penetration into the adaxialsurface greater. In darkness the rates of penetration over 24 h remain constantor fall but slightly for all species. Light consistently promotespenetration but with Beta there is a lag before entry is acceleratedinto the abaxial surface as has previously been reported foryoung primary leaves of Phaseolus. For the remaining speciesthe courses of penetration in both light and darkness into bothsurfaces follow similar patterns. As the light intensity isincreased entry is enhanced but the limit of response variesbetween species, between surfaces within species, and in trifoliateleaves of Phaseolus with age. For the six species the order of the relative rates of entryis closely similar whether comparisons are made in light ordarkness or between abaxial and adaxial surfaces: viz. Zea >Helianthus > Phaseolus (primary) > Phaseolus (trifoliate)> Pisum = Beta = Gossypium. The observed specific differencesare discussed in relation to variations in leaf structure, theproperties and thickness of the cuticle and the physiologicaland metabolic processes which influence transport within theepidermal tissues after it has passed through the cuticle bydiffusion.