Stomatal density and aperture in non-vascular land plants are non-responsive to above-ambient atmospheric CO2 concentrations

Background and Aims Following the consensus view for unitary origin and conserved function of stomata across over 400 million years of land plant evolution, stomatal abundance has been widely used to reconstruct palaeo-atmospheric environments. However, the responsiveness of stomata in mosses and hornworts, the most basal stomate lineages of extant land plants, has received relatively little attention. This study aimed to redress this imbalance and provide the first direct evidence of bryophyte stomatal responsiveness to atmospheric CO₂.Methods A selection of hornwort (Anthoceros punctatus, Phaeoceros laevis) and moss (Polytrichum juniperinum, Mnium hornum, Funaria hygrometrica) sporophytes with contrasting stomatal morphologies were grown under different atmospheric CO₂ concentrations ([CO₂]) representing both modern (440 p.p.m. CO₂) and ancient (1500 p.p.m. CO₂) atmospheres. Upon sporophyte maturation, stomata from each bryophyte species were imaged, measured and quantified.Key Results Densities and dimensions were unaffected by changes in [CO₂], other than a slight increase in stomatal density in Funaria and abnormalities in Polytrichum stomata under elevated [CO₂].Conclusions The changes to stomata in Funaria and Polytrichum are attributed to differential growth of the sporophytes rather than stomata-specific responses. The absence of responses to changes in [CO₂] in bryophytes is in line with findings previously reported in other early lineages of vascular plants. These findings strengthen the hypothesis of an incremental acquisition of stomatal regulatory processes through land plant evolution and urge considerable caution in using stomatal densities as proxies for paleo-atmospheric CO₂ concentrations.     

COMPARATIVE ULTRASTRUCTURAL STUDIES OF SPERMATOGENESIS IN THE METZGERIALES (HEPATICAE). I. THE BLEPHAROPLAST OF PALLAVICINIA LYELLII

The blepharoplast in the young spermatid of Pallavicinia is similar to that of other hepatics in that it comprises a four-layered multilayered structure (MLS) and two staggered, dimorphic basal bodies. The spline, approximately 40 μm in length and extending through nearly two full gyres, comprises 20 parallel microtubules at its anterior end and narrows to 17 at the posterior limit of the subjacent lamellar strip (LS). Behind this, the spline shank, approximately 32 μm in length, is reduced to six tubules. The LS curves around the spermatid, following the anterior one-third of the first gyre of the spine, and is approximately 7.5 μm in length, the longest yet recorded for the bryophytes. It is spatulate in outline, equaling the width of the spline anteriorly but tapering steeply from the right-hand side behind the anterior basal body (ABB). It then extends posteriorly as a narrow strip beneath the left-hand margin of the spline. The basal bodies of the greatly staggered flagella are nonoverlapping and separated by a distance of about 4.4 μm. The subapical ABB and PBB measure (including the ventral triplet extensions and transition zones) 1.2 μm and 2.4 μm in length, respectively. A short, narrow aperture equaling one tubule-diameter in width is located in the spline directly beneath the ABB. The anterior mitochondrion is about 7 μm long and follows the outline of the overlying LS, while a cupshaped posterior mitochondrion is appressed to the plastid. Comparisons with other taxa indicate that major distinguishing features of metzgerialian blepharoplasts are highly staggered, nonoverlapping basal bodies, greatly elongate anterior mitochondria, and six-tubule shanks. Great differences between the spermatids suggest wide phylogenetic discontinuities between the genera of the Metzgeriales.     

Thallus Differentiation in the Marchantialean Liverwort Asterella wilmsii (Steph.) with Particular Reference to Longitudinal Arrays of Endoplasmic Microtubules in the Inner Cells

This light and electron microscope study of the liverwort Asterellareveals that, as in other Marchantiales, the cells lining thedorsal air chambers are highly vacuolate with numerous amylochloroplastsin the peripheral cytoplasm. The ventral parenchyma in the midribof the thallus contains aseptate fungal hyphae surrounded byan interfacial matrix and host cytoplasm forming transvacuolarstrands. These are lined with microtubules, rarely seen in otherfungal-hepatic associations or in mycorrhizas. Numerous lipidbodies found in all the thallus cells are thought to be associatedwith perennation during the winter dry season. Elongated, thick walled inner thallus cells, between the dorsalair chambers and the fungus-containing tissue, have a cytologicalorganization not previously recorded in land plants. Initiallyhighly vacuolate, with numerous microtubules of random orientationlining the tonoplast, these cells subsequently show interdigitationof vacuoles and cytoplasm producing a labyrinth of sphericaland elongate tonoplast profiles lined by longitudinal arraysof microtubules. At the same time the cytoplasm becomes increasinglyelectron-lucent and the ribosomes, progressively lost from theER, clump together. At maturity the inner thallus cells arehighly polarized with most of the vacuoles lying nearer thethallus apex. In pits in the end walls, numerous plasmodesmata,with expanded cytoplasmic annuli recall the plamodesmatal fieldsin the mesophyll and phloem of the leaves in vascular plants. Far from being supporting parenchyma or sclerenchyma as assumedhitherto, the inner thallus cells of Asterella are clearly highlydifferentiated. Their vacuole microtubule associations are highlysuggestive of a microtubule-based translocation system akinto that seen in many animal cells and perhaps fungal hyphae,but very different from bulk flow in sieve elements and actin-basedcytoplasmic streaming.Copyright 1994, 1999 Academic Press Cytoskeleton, liverwort, microtubules, plasmodesmata, solute transport     

The structure of the Holliday junction, and its resolution

The Holliday (four-way) junction is a critical intermediate in homologous genetic recombination. We have studied the structure of a series of four-way junctions, constructed by hybridization of four 80 nucleotide synthetic oligonucleotides. These molecules migrate anomalously slowly in gel electrophoresis. Each arm of any junction could be selectively shortened by cleavage at a unique restriction site, and we have studied the relative gel mobilities of species in which two arms were cleaved. The pattern of fragments observed argues strongly for a structure with two-fold symmetry, based on an X shape, the long arms of which are made from pairwise colinear association of helical arms. The choice of partners is governed by the base sequence at the junction, allowing a potential isomerization between equivalent structural forms. Resolvase enzymes can distinguish between these structures, and the resolution products are determined by the structure adopted, i.e., by the sequence at the junction. In the absence of cations, the helical arms of the junction are fully extended in a square configuration, and unstacking results in junction thymines becoming reactive to osmium tetroxide.     

Effects of de- and rehydration on food-conducting cells in the moss Polytrichum formosum: a cytological study

BACKGROUND AND AIMS: Moss food-conducting cells (leptoids and specialized parenchyma cells) have a highly distinctive cytology characterized by a polarized cytoplasmic organization and longitudinal alignment of plastids, mitochondria, endoplasmic reticulum and vesicles along endoplasmic microtubules. Previous studies on the desiccation biology of mosses have focused almost exclusively on photosynthetic tissues; the effects of desiccation on food-conducting cells are unknown. Reported here is a cytological study of the effects of de- and rehydration on food-conducting cells in the desiccation-tolerant moss Polytrichum formosum aimed at exploring whether the remarkable subcellular organization of these cells is related to the ability of mosses to survive desiccation. METHODS: Shoots of Polytrichum formosum were dehydrated under natural conditions and prepared for transmission and scanning electron microscopy using both standard and anhydrous chemical fixation protocols. Replicate samples were then fixed at intervals over a 24-h period following rehydration in either water or in a 10 microM solution of the microtubule-disrupting drug oryzalin. KEY RESULTS: Desiccation causes dramatic changes; the endoplasmic microtubules disappear; the nucleus, mitochondria and plastids become rounded and the longitudinal alignment of the organelles is lost, though cytoplasmic polarity is in part retained. Prominent stacks of endoplasmic reticulum, typical of the hydrated condition, are replaced with membranous tubules arranged at right angles to the main cellular axis. The internal cytoplasm becomes filled with small vacuoles and the plasmalemma forms labyrinthine tubular extensions outlining newly deposited ingrowths of cell wall material. Whereas plasmodesmata in meristematic cells at the shoot apex and in stem parenchyma cells appear to be unaffected by dehydration, those in leptoids become plugged with electron-opaque material. Starch deposits in parenchyma cells adjoining leptoids are depleted in desiccated plants. Rehydration sees complete reestablishment over a 12- to 24-h period of the cytology seen in the control plants. Oryzalin effectively prevents leptoid recovery. CONCLUSIONS: The results point to a key role of the microtubular cytoskeleton in the rapid re-establishment of the elaborate cytoplasmic architecture of leptoids during rehydration. The reassembly of the endoplasmic microtubule system appears to dictate the time frame for the recovery process. The failure of leptoids to recover normal cytology in the presence of oryzalin further underlines the key role of the microtubules in the control of leptoid cytological organization.     

The crystal structure of Neisseria gonorrhoeae PriB reveals mechanistic differences among bacterial DNA replication restart pathways

Reactivation of repaired DNA replication forks is essential for complete duplication of bacterial genomes. However, not all bacteria encode homologs of the well-studied Escherichia coli DNA replication restart primosome proteins, suggesting that there might be distinct mechanistic differences among DNA replication restart pathways in diverse bacteria. Since reactivation of repaired DNA replication forks requires coordinated DNA and protein binding by DNA replication restart primosome proteins, we determined the crystal structure of Neisseria gonorrhoeae PriB at 2.7 Å resolution and investigated its ability to physically interact with DNA and PriA helicase. Comparison of the crystal structures of PriB from N. gonorrhoeae and E. coli reveals a well-conserved homodimeric structure consisting of two oligosaccharide/oligonucleotide-binding (OB) folds. In spite of their overall structural similarity, there is significant species variation in the type and distribution of surface amino acid residues. This correlates with striking differences in the affinity with which each PriB homolog binds single-stranded DNA and PriA helicase. These results provide evidence that mechanisms of DNA replication restart are not identical across diverse species and that these pathways have likely become specialized to meet the needs of individual organisms.     

Adjacent 2 meiotic disjunction. Report of a case resulting from a familial 13q;15q balanced reciprocal translocation and review of the literature

Summary An abnormal short-lived female infant with almost complete trisomy 13 (pterq32 or 33) and partial monosomy 15 (pterq14 or 15) resulting from an adjacent 2 meiotic disjunction of a paternal reciprocal translocation is described. Cases with monosomy of chromosome 15 material are reviewed. It appears likely that monosomy of an interstitial long arm segment, approximating to 15q2124, imparts the lethality associated with the full monosomic condition. Adjacent 2 disjunction in man has been further characterised by reviewing the literature.