Models of the Use of Root-zone CO2 by Selected North American Isoetids

Sediment CO₂, entering via the roots, contributes a significantportion of the total carbon uptake for isoetids (small, evergreen,submersed, vascular plants). Laboratory studies of inorganiccarbon uptake via the roots and shoots by five isoetids wereused to model the use of root-zone CO₂. Simple first-order linearmodels accounted for at least 75 per cent of the variation inthe data for Gratiola aurea, Isoetes macrospora, Littorellauniflora and Lobelia dortmanna. For Eriocaulon septangulare,which relies almost exclusively on root-zone CO₂, models couldaccount for only about 62 per cent of the variation in root-zoneCO₂ use. For each species, we present the best fitting regressionof root-zone CO₂ use as a function of root- and shoot-zone CO₂concentrations. For the species studied, carbon uptake was not saturated atfield concentrations of root and shoot-zone CO₂. Maximum ratesof carbon uptake were lower for species that naturally occurredat greater depths, compared with species more common in shallowwater. At equal external CO₂ concentrations carbon entry perunit root surface area was several times more rapid than entryper unit shoot surface area for L. dortmanna. The entry ratesper unit root and shoot surface area were about equal for G.aurea and E. septangulare. Shoots were equally or more permeablethan the roots of L. uniflora and I. macrospora, a fact thatmay be related to the functioning of crassulacean acid metabolismin these plants. Carbon, CO₂, photosynthesis, isoetid, Eriocaulon septangulare, Gratiola aurea, Isoetes macrospora, Littorella uniflora, Lobelia dortmanna     

Utilization of Sediment CO2 by Selected North American Isoetids

The photosynthetic uptake of root-zone CO₂ was determined forEriocaulon septangulare, Gratiola aurea, Isoetes macrospora,Littorella uniflora var. americana and Lobelia dortmanna aspart of a study of the photosynthetic carbon economy of submergedaquatic isoetids. The pH and dissolved inorganic carbon (DIC)of the sediment interstitial water in four Wisconsin lakes reflectedthe water column character, where the DIC increased with depthin the sediment to concentrations five to ten times those ofthe water column. Sediment free CO₂ concentrations were 5–50times those in the water column and were similar at all sites(about 05–1.0mM CO₂ in the root-zone). In ‘pH-drift’studies these plants were unable to take up HCO₂^–. Laboratory determinations of the carbon uptake from the rootand shoot-zones were made for all five species. These experimentsshowed that CO₂ in the root-zone accounted for 65–95 percent of external carbon uptake for the five species. For G.aurea and E. septangulare, root-zone CO₂ was > 85 per centof carbon uptake. Carbon, CO₂, photosynthesis, sediment, isoetid, Eriocaulon septangulare, Gratiola aurea, Isoetes macrospora, Littorella uniflora, Lobelia dortmanna     

Studies of Diurnal Acid Fluctuations in British Isoetid-type Submerged Aquatic Macrophytes

British isoetid species are examined for the presence of diurnalfluctuations in tritratable acidity (to pH 6·4), in plantscollected directly from a small lake and in plants grown inconstant conditions in the laboratory. Wide diurnal fluctuationsare present in Isoetes lacustris and in both submerged and terrestrialpopulations of Littorella uniflora. They are absent in Lobeliadortmanna, Subularia aquatica, Eriocaulon septangulare, Ranunculusflammula and Pilularia globulifera. The significance of submerged CAM is discussed in relation toother carbon accumulating mechanisms in isoetids and in considerationof their general ecology. Crassulacean acid metabolism, photosynthesis, isoetid, oligotrophic lakes     

Root Cap Structure in Isoetes macrospora Dur

Root meristem cells of Isoetes macrosporausually have one plastidwhich is associated with the prominent nucleus, numerous ribosomesand mitochondria, and small vacuoles. During mitosis each plastidappears to replicate so that each daughter cell contains oneplastid. The cell walls of the meristem cells are traversedby numerous plasmodesmata. Central cells of the root cap lackdistally displaced plastids but have one or more amyloplastsassociated with the nucleus. These cells also contain largeprotein deposits. Peripheral root cap cells are characterizedby being vacuolated, and by possessing a few dictyosomes andprotein deposits. They appear to be sloughed infrequently. Isoetes macrospora Dur, root cap, protein bodies, ultrastructure     

The quillworts (Isoetes) of India: distribution, endemism and species radiation

The genus Isoetes L. in India is represented by 14species, of which eight species are recognized as being local endemics [confinedto one particular phyto-geographical division (PGD)] while the remaining sixoccur in more than one PGD and are described as endemic to a wider range. Thelocal endemic species are Isoetes dixitei,Isoetes panchganiensis and Isoetessahyadriensis in Western Ghats region; Isoetespantii, Isoetes bilaspurensis, Isoetesreticulata and Isoetes tuberculata inChotanagpur Malwa Vindhya Plateau and Isoetes debii innortheastern India. The wider endemic species are Isoetespanchananii, Isoetes sampathkumaranii,Isoetes rajasthanensis, Isoetesmahadevensis, Isoetes indica andIsoetes coromandelina. Our studies on the patterns ofendemism suggest that the radiation of quillworts advanced from dry lowlandareas to the rainy uplands and mountains. Isoetescoromandelina is the first Indian quillwort to colonize in lowlandsof the coastal zone (Coromandel), from where it spread to different parts of thesub-continent and gave rise to new species. Thus this species is a key Indianspecies which has played an important role in the radiation of quillwort in thecountry and appeared as the connecting link among the quillwort flora of variousPGDs. The centres of diversity for almost all the presently known Indianquillworts species are recognized.     

Ultrastructure, Origin, and Composition of the Protein Bodies in the Ligule of Isoetes lacustris L.

The basal cells in the ligule of Isoetes lacustris contain numerousprotein bodies, the contents of which can be digested enzymicallyby pronase and are stained red by treatment with ninhydrin Schiff'sreagent. Two types of protein bodies can be distinguished ultrastructurally:spherically-shaped bodies with granular contents and spindle-likebodies with fibrillar contents. Both are ensheathed by singlemembranes and do not show any solid inclusions within theirmatrix. The protein bodies probably arise from dilatation of the endoplasmicreticulum (ER) cisternae. This conclusion is based upon threeobservations: (a) The protein bodies occasionally show membranecontinuity with the ER; (b) ribosomes and polysomes are frequentlyattached to the protein-body membranes; (c) the contents ofthe protein bodies and of the dilated ER cisternae show similarultrastructural features. The dilatation of the ER cisternae is assumed to be a resultof protein accumulation in the intracisternal space. Based upon the results of polyacrylamide gel electrophoresis,it is likely that the spherically-shaped protein bodies storepredominately two proteins with molecular weights of 51300 and55800 D, while the spindle-like bodies store two proteins withmolecular weights of 92000 and 98000 D. The results presented do not permit a definite conclusion regardingthe function of the ligule of Isoetes lacustris but it is suggestedthat it may have a nutritive role. Isoetes lacustris L., ligule, protein bodies, endoplasmic reticulum, ultrastructure     

Activities of Carboxylation Enzymes in Freshwater Macrophytes

Fifteen species of freshwater macrophytes, mainly from cool,temperate waters, were assayed for ribulose bisphosphate carboxylase-oxygenase(RuBPCase) and phosphoenolpyruvate carboxylase (PEPCase) activities.In extracts from all the species RuBPCase was the most activecarboxylation enzyme, and the RuBPCase/PEPCase ratio was atleast 2·0, even for the submersed species Isoetes lacustrisL. and Littorella unifiora (L.) Aschers. which have been reportedto show Crassulacean Acid Metabolism (CAM) activity. The PEPCaseactivity in I.lacustris was lower than that found in some non-CAM-likespecies. In this respect, I.lacustris and L unifiora differfrom most terrestrial CAM plants. However, these two species,along with Potamogeton praelongus Wulf. and Juncus bulbosusvar.fluitans L., had the lowest RuBPCASE/PEPCase ratios, lowerthan found in terrestrial C₃ species; suggesting that the potentialfor substantial photosynthetic metabolism of C₄ acids existsin some temperate, submersed plants. In the three amphibiousspecies (Potamogeton polygonifolius Pourr., Mentha aquaticaL., and Hippuris vulgaris L.) examined, the aerial leaves exhibitedhigher RuBPCase activities than the submersed leaves. Key words: Ribulose bisphosphate carboxylase-oxygenase, phosphoenolpruvate carboxylase, freshwater macrophytes     

Investigation of the Presence of Phenolic Compounds in Monocotyledonous Cell Walls, using UV Fluorescence Microscopy

Harris and Hartley (1976, 1980) demonstrated the presence offerulic acid in cell walls of certain monocotyledons using UVfluorescence microscopy (fluorescing green after treatment withammonium hydroxide solution). The presence or absence of thistype of fluorescence is apparently critical in higher levelsystematics of monocotyledons. In order to evaluate the significanceof this character, cell wall fluorescence was investigated ina range of monocotyledon species, particularly the AustralianXanthorrhoeaceae sensu lato (Bedford et al., 1986), which werenot investigated in earlier studies. This family is widely regardedas polyphyletic and was divided into several families by Dahlgren,Clifford and Yeo (1985). Some of its constituent genera, suchas Dasypogon, Kingia and Calectasia, have been linked with bothcommelinoid and non-commelinoid monocotyledons, and are of obscureaffinity. Some genera of Xanthorrhoeaceae sensu lato (Baxteria,Calectasia, Dasypogon and Kingia) show this type of green cellwall fluorescence and may therefore be more closely linked withthe commelinoid monocotyledons, rather than the Lilianae-Asparagales,as previously placed (Dahlgren et al., 1985).Copyright 1994,1999 Academic Press Asparagales, Dasypogonaceae, fluorescence, Hanguana, monocotyledons, systematics, Xanthorrhoeaceae     

Endodermis-like Sheaths in the Submerged Freshwater MacrophyteRanunculus trichophyllusChaix

Light, fluorescence and electron microscopical analysis of therooted freshwater plantRanunculus trichophyllusrevealed a peculiaranatomical feature. In addition to the true endodermis encirclingthe root stele, endodermis-like sheaths occurred around eachvascular bundle of the leaf segments and of the eustelic stemwith its large central cavity, which assumed an anatomical featureresembling that of some pteridophyte stems. These impermeablesheaths, whose cells differentiate suberized walls, can playa major role in hampering the apoplastic leakage of the pressurizedwater solution which flows throughout the plant in xylem vesselsand contains the mineral nutrients taken up by the roots fromthe sediment. Moreover, these sheaths can function in preventingflooding of the aerenchymatic cavities of the submerged organs.In this way the endodermis-like sheaths preserve the correctcirculation of gas and nutrient solution through the entireorganism and assume great significance as a structural mechanismevolved by this species to survive and grow underwater.Copyright1999 Annals of Botany Company. Ranunculus trichophyllus,freshwater macrophyte, submerged angiosperm, anatomy, endodermis, endodermis-like sheaths, light microscopy, fluorescence microscopy.     

Epicuticular Phenolics Over Guard Cells: Exploitation for in situ Stomatal Counting by Fluorescence Microscopy and Combined Image Analysis

Guard cells emit an alkali-induced, blue fluorescence upon excitationby ultraviolet radiation (emission maximum energy at 365 nm).Fluorescence emission of guard cells was brighter than thatof the neighbouring epidermal cells in a number of wild andcultivated plants including conifers, but the relative fluorescenceintensity and quality was species-dependent. Three representativeplants possessing stomatal complexes which differed morphologicallywere studied: Olea europaea, Vicia faba and Triticum aestivum.Immersing leaves of these plants in chloroform for 30 s (therebyremoving epicuticular waxes) significantly reduced the intensityof the fluorescence emitted by guard cells. This indicates thatguard cell fluorescence could be due to either an increasedconcentration of fluorescing compounds (probably wax-bound phenolics),or a thicker cuticular layer covering the guard cells. Giventhat the alkali-induced blue fluorescence of the guard cellsis a common characteristic of all plants examined, it couldbe used as a rapid and convenient method for in situ measurementsof the number, distribution and size of stomatal complexes.The proposed experimental procedure includes a single coatingof the leaf surface by, or immersion of the whole leaf in, a10% solution of KOH for 2 min, washing with distilled water,and direct observation of the leaf surface under the fluorescencemicroscope. Fluorescence images were suitable for digital imageanalysis and methodology was developed for stomatal countingusing Olea europaea as a model species. Copyright 2001 Annalsof Botany Company Cuticle, epicuticular waxes, fluorescence microscopy, image analysis, phenolics, stomata     

Nuclear DNA Content in Twelve Species of Alstroemeria L. and Some of their Hybrids

Nuclear DNA content (2C-value), estimated through flow cytometryusing propidium iodide (PI), was shown to vary from 36.5 pgto 78.9 pg among 29 accessions of 12Alstroemeria species (2n=2x =16). The extremes were found inA. magnifica ssp.magnificaand inA. ligtu ssp.simsii , both belonging to the Chilean speciesgroup. The four Brazilian species exhibited less variation innuclear DNA content (49.8–56.4 pg), than the eight Chileanspecies (36.5–78.9 pg). Nuclear DNA content was positivelycorrelated (r =0.92,n =7,P <0.01) with the total chromosomelength. It was also positively correlated (r =0.85,n =5,P <0.01)with the length of C-bands, when only the Chilean species wereconsidered. When both karyotype parameters, length of non-C-bandedchromosome regions (x) and length of C-bands (y) were determined,it was possible to predict the nuclear DNA content (z) withthe formula z=0.65x +1.31y-0.45 (R ²=0.97,P =0.004). The DAPI fluorescence of most accessions was proportional tothe PI fluorescence (r =0.98,P <0.001), except for one accessionofA. ligtu , that had a relatively high PI/DAPI ratio (1.88).The PI/DAPI ratios of the Brazilian species were lower (1.59–1.67)than those of the Chilean species (1.68–1.88), which mightreflect a difference in base pair composition. Four groups ofspecies could be distinguished on the basis of fluorescencevalues. Diploid interspecific hybrids were shown to have a DNAcontent intermediate to the values of the parents involved.Both the PI and the DAPI fluorescence values of these hybridsapproximated the mid parent values. Tetraploids, derived fromselfing of diploids, had PI and DAPI fluorescence values thatwere twice that of the diploid hybrids. It was possible to distinguishaneuploids from euploids based on fluorescence values. Alstroemeria ; aneuploidy; C-banding; DAPI; evolution; flow cytometry; genome size; geophytes; karyotypes; Inca Lily; nuclear DNA; propidium iodide     

Phytoplankton monitoring by flow cytometry

The application of flow cytometry to the monitoring of phytoplanktonis demonstrated. A comparison is made with conventional approachesto phytoplankton monitoring: light microscopy for the determinationof species abundance, and chlorophyll a determination and insitu chlorophyll a measurement by fluorescence for the determinationof the biomass. Flow cytometric measurements correlate wellwith these conventional types of measurements, as has been shownby comparing a full year of monitoring data obtained at a fixedmonitoring location 10 km off the Dutch coast. Flow cytometrybridges the gap between labour-intensive, but highly informative,microscopic observations and simple biomass measurements withless information content: via flow cytometry optical data areobtained at high speed for individual particles, which can betranslated into biomass information. On the basis of the flowcytometric measurements, rough discrimination of phytoplanktonspecies groups is possible, particularly for the abundant species.Of crucial importance is careful calibration of the flow cytometer,to ensure quantitative and comparable measurements over a longperiod of time. Calibration and quality assurance aspects arecovered in detail. ³Present address: Akzo Nobel Central Research Laboratories Arnhem,Department CRL, PO Box 9300, NL-6800 SB Arnhem, The Netherlands     

Comparative Embryo Sac Morphology at Anthesis of Cultivated and Wild Species of Arachis

Introgression of germplasm from wild to cultivated species ofArachis is severely impeded because abortion processes oftenoccur as a prepeg-, peg-(gynophore), or postpeg-elongation event.A comparative study of embryo sac morphology at anthesis wasundertaken to determine if observable differences were presentthat could possibly explain abortion prior to peg tip swellingfollowing soil penetration. Two wild Arachis species (A. duranensisand A. stenosperma) plus A. hypogaea cultivars NC 6 and Argentinewere studied. Differences in starch grain concentration andcytoplasmic stranding organization were observed between A.hypogaea cultivars and the wild Arachis species. These differencesprobably have a significant impact on energy availability atsyngamy and the subsequent early cell division of the embryo.An improper energy balance could contribute to the onset ofabortion in interspecific hybrids. Modification in egg apparatusorganization among all species was also observed which may accountfor low percentages of seed recovery resulting from interspecifichybridization attempts. Embryo sac morphology, interspecific hybridization, Arachis hypogaea L., Arachis species, light microscopy, scanning electron microscopy, embryo abortion, fertilization incompatibility, peanuts, groundnut     

Integrating conventional microscopy and molecular analysis to analyse the abundance and distribution of four Calanus congeners in the North Atlantic

Analysis of the demographic structure of Calanus species inthe North Atlantic presents particular difficulties due to theoverlapping spatial distributions of four main congeneric species(Calanus finmarchicus, Calanus helgolandicus, Calanus glacialisand Calanus hyperboreus). These species have similar morphologies,making microscopic discrimination only possible between someof the species at late copepodite or adult stages. However,molecular techniques now offer the possibility of screeningsignificant numbers of specimens and unambiguously identifyingthem to species, regardless of developmental stage. Unfortunately,the processing rate of specimens by molecular methods is stilltoo low to offer a realistic alternative to microscopy for analysisof samples from large field surveys. Here, we outline and testan approach involving the use of molecular methodology in conjunctionwith conventional microscopy to assess the species assignmentof developmental stage abundances of Calanus congeners. Ourstudy has highlighted many important methodological issues.First, it cannot be assumed that the species composition ishomogeneous across the development stages; applying proportionalspecies composition of adults to morphologically undistinguishableearlier development stages can result in error. The second importantconclusion is that prosome length may be a highly unreliablediscriminator of C. finmarchicus and C. glacialis.     

Structure and Development of Sieve Elements in the Root of Isoetes muricata Dur.

Root tissues of Isoetes muricata Dur. were fixed in glutaraldehydeand postfixed in osmium tetroxide for electron microscopy. Veryyoung root sieve elements can be distinguished from contiguousparenchyma cells by the presence of crystalline and/or fibrillarproteinaceous material in dilated cisternae of rough endoplasmicreticulum (ER). Similar crystalline-fibrillar material accumulatesin the perinuclear space. During differentiation, the portionsof ER enclosing this proteinaceous substance become smooth surfacedand migrate to the cell wall. Along the way many of them formmultivesicular bodies which fuse with the plasmalemma, dischargingtheir contents toward the wall. Nuclear degeneration is pycnotic.At maturity, the sieve element contains a degenerate, filiformnucleus, plastids, and mitochondria. In addition, the wall ofthe mature sieve element is lined by a plasmalemma and a parietalnetwork of smooth ER. Sieve-area pores are present in both endand lateral walls of mature sieve elements. Whereas a singlecluster of pores occurs in each end wall, the pores of the lateralwalls are solitary and few in number.     

Variation in the DNA Content of Glycine Species

Nuclei were isolated from cotyledons of a range of accessionsfrom 14 species of Glycine. These were stained with ethidiumbromide and the relative fluorescence for each genotype wasmeasured by flow cytometry. The DNA content was estimated bycomparison of relative fluorescence with that from nuclei fromseedling leaves of Allium cepa, whose DNA content has been calculatedpreviously by chemical assay. The 4C amounts for diploid Glycineranged from 3.80 to 6.59 pg. Two groups of diploid species appearedfrom the analysis. The first consisted of species with amountsranging from 3.80 to 5.16 pg and included G. canescens (AA),G. argyrea (A₁ A₁), G. clandestina (A²A²), G. microphylla(BB),G. latifolia (B₁B₁), G. tabacina 2n=40 (B²B²), G. tomentella2n=38 (EE) and 2n=40 (DD), G. max and G. soja (GG), G. arenariaand G. latrobeana. A second group had higher DNA contents rangingfrom 5.27 to 6.59 pg, and consisted of G. curvata, G. cyrtoloba(CC), and G. falcata (FF). The polyploid species, G. tabacina2n=80 (AABB, BBB¹B¹), G. tomentella 2n=78 and 2n=80 (AAEE andDDEE, respectively) contained amounts approximating to the sumsof the respective parental diploid species thought to have givenrise to these allotetraploids. Intraspecific variation was detectedin the DNA content of G. canescens. Within the overall distributionof DNA amounts found in A genome species, each genome containeda range of DNA contents specific to that species. This phenomenonwas also detected amongst B genome species.     

Micromorphological approach to the systematics of Mediterranean Isoëtes species (Isoëtaceae,Lycopodiophyta): analysis of the megaspore surface

Megaspore ornamentation is one of the few morphological characters used in the taxonomy of the genus Isoëtes. In the present work, we test the application of this character for distinguishing some Isoëtes species occurring in the Mediterranean: Isoëtes sicula, which according to some authors should be included as a variety in I. histrix, and the recently described I. todaroana, whose affinity with other taxa is currently unknown. Two additional species (I. duriei, I. velata) were also included in the analysis. The megaspores were studied using scanning electron microscopy combined with energy-dispersive X-ray spectroscopy and chemical treatment. In all species, the megaspore surface is covered by a siliceous coating; removal of this coating reveals an underlying three-dimensional network of fused rodlets. A unique pattern of this network occurs in I. sicula, supporting its separation from I. histrix. Comparison between the patterns observed in the examined Mediterranean species shows that I. todaroana is most similar to I. histrix.     

The Ligule of Isoetes lacustris: Ultrastructure, Mucilage Composition, and a Possible Pathway of Secretion

Mature ligules of Isoetes lacustris can be divided anatomicallyinto three ultrastructurally different regions. First, the basalregion contains large numbers of two types of protein bodies.Second, the cells of the sub-marginal region are characterizedby a well developed Golgi apparatus closely associated withthe tubular compartments of the endoplasmic reticulum. Third,the peripheral region consists of one to three layers of cellsshowing an extremely well developed rough asternal and smoothtubular endoplasmic reticulum (ER). The tubular ER compartmentsare frequently observed in close attachment to the plasmalemma.The outermost peripheral cells are covered with a mucilaginouslayer. The dry matter in the mucilage consists of 49 per centpolysaccharides and 22 per cent proteins. The polysaccharidefraction, analysed by ion exchange chromato-graphy, consistsmainly of glucose, arabinose, galactose and uronic acids. Theprotein fraction was analysed by SDS gel dectrophoresis andby high performance liquid-chromatographic separation of theamino acids. The analysis shows a protein pattern very similarto that of the peripheral ligule tissue. It is suggested, therefore,that the material of the external mucilage is secreted by theperipheral ligule cells. The secretional mechanism appears tobe a direct release of polysaccharides and proteins by the tubularcomponents of the ER. There is no indication of secretion viathe Golgi apparatus. Because of its high activity in proteinsynthesis and secretion, it is suggested that the ligule isa vestigial structure, which, in extinct genera, might havefunctioned as a digestive organ. Isoetes lacustris, endoplasmic reticulum, ligule, ultrastructure, Polysaccharide secretion, protein secretion     

Effect of High Temperature During Grain-filling on the Structure of Developing and Malted Barley Grains

High temperatures (up to 35 °C) were applied to plants ofmalting barley,Hordeum vulgareL. (‘Schooner’) fora period of 5 d during grain-filling. Heat treatment had a profoundeffect on the structure of the mature barley grain. There wasevidence of degradation of endosperm storage products in heat-treatedgrain. Starch granule development was reduced in sub-aleuronecells following heat treatment and alterations to starch granuledistribution and growth were observed in the endosperms of thesegrains. Endosperm cell wall and crushed cell layer (CCL) developmentwere sensitive to high temperatures, with the reduced thicknessof the CCL and generally patchy Calcofluor fluorescence of endospermcell walls indicative of partial hydrolysis of ß-glucans.Increased growth of the embryo took place in heat-treated grainscompared with control grains. Endosperm texture was generallymore friable in heat-treated grains than in control grains,and these grains overmodified during malting, with considerabledegradation of starch in the form of extensive pitting of A-typestarch granules. Evidence is presented for developmental andgermination events occurring simultaneously within the developinggrain.Copyright 1998 Annals of Botany Company Barley,Hordeum vulgareL., starch granules, crushed cell layer, scutellum, embryo, fluorescence microscopy, scanning electron microscopy, confocal microscopy, malting quality.     

MEGASPORE WALL ULTRASTRUCTURE IN ISOETES

In Selaginella, megaspore wall ultrastructure (unit morphology and arrangement) is correlated with taxonomic position. In some Pennsylvanian lycopods there is a correlation with dispersal strategy. This study was designed to assess any correlation between habitat preference and wall ultrastructure in Isoetes. Except for a few minor structural correlations, wall ultrastructure appears to be constant in terrestrial, amphibious, and aquatic species of Isoetes. Several distinct sizes of megaspores occur in some megasporangia and correspond to 1) full-size megaspores with siliceous coatings, 2) small megaspores with siliceous coatings, and 3) full-size megaspores without siliceous coatings. The uniformity in wall ultrastructure within modem Isoetes together with the lack of uniformity of fossil isoetalean megaspore wall ultrastructure suggest that the modem species of Isoetes are closely related.