Orientation of Wall Microfibrils in Avena Coleoptiles and Mesocotyls and in Pisum Epicotyls

Microfibrils (MFs) on the inner surface of the walls of Avenacoleoptile and mesocotyl cells and of Pisum epicotyl cells wereexamined by a replica method. In the elongating epidermis ofthese three organs, cells having MFs that were transverse, obliqueor longitudinal to the elongation axis were intermingled. Inthe elongating parenchymal tissues, all cells deposited MFstransversely. In non-elongating cells of Avena coleoptiles andPisum epicotyls, the orientation of MFs on the inner wall surfaceof both epidermal and parenchymal cells was more longitudinalthan in elongating cells. These observations on the orientationsof MFs are compatible with those our previously reported observationson the orientations of microtubules (MT) (Iwata and Hogetsu1988). Disruption of MTs of Avena coleoptiles by treatment withamiprophosmethyl caused changes in the orientation of depositionof MFs. These results support the idea that MFs are usuallyco-aligned with MTs in organ cells and that the orientationof MFs is controlled by MTs. The averaged direction of MFs, visualized under polarized light,showed a clear difference between the epidermal and inner-tissuecell walls in the elongating regions of the three organs. Inalmost all elongating and non-elongating epidermal cells, theaveraged direction of MFs was longitudinal, while it was transversein all inner-tissue cells. (Received December 16, 1988; Accepted April 28, 1989)     

Specific arrangements of cortical microtubules are correlated with the architecture ofmeristems in shoot apices of angiosperms and gymnosperms

Arrangements of corticalmicrotubules (MTs) as seen in median longitudinal cryosections of shoot apices of several angiosperms and gymnosperms were studied by indirect immunofluorescence microscopy.Bryophyllum, Clethra, Helianthus, Houttuynia, Vinca (angiosperms), andPinus, Cedrus, Cedrus andGinkgo (gymnosperms) were examined. In all angiosperm apices collected during the growing season, MTs were mainly arranged anticlinally in the tunica, randomly in the corpus, and transversely in the rib meristem. This pattern of arrangements of MTs was further confirmed by electron microscopy inBryophyllum apices. In the apices of winter shoots MTs in the rib meristem were arranged randomly, indicating a seasonal change with respect to their arrangment. In all examined gymnosperm apices, populations of superficial cells showed both random and anticlinal arrangements of MTs, in contrast to those of angiosperm apices that consistently show anticlinally arranged MTs. In the shoot apices of both angiosperms and gymnosperms, cortical MTs were arranged perpendicularly to the directions of cell expansion. The significance of MTs in the maintrnance of the different architectures of shoot apices in angiosperms and gymnosperms is discussed.     

Mechanism for formation of the secondary wall thickening in tracheary elements: Microtubules and microfibrils of tracheary elements of Pisum sativum L. and Commelina communis L. and the effects of amiprophosmethyl

Arrangements of microfibrils (MFs) and microtubules (MTs) were examined in tracheary elements (TEs) of Pisum sativum L. and Commelina communis L. by production of replicas of cryo-sections, and by immunofluorescence microscopy, respectively. The secondary wall thickenings of TEs of Pisum and Commelina roots have pitted and latticed patterns, respectively. Most MFs in the pitted thickening of Pisum TEs retain a parallel alignment as they pass around the periphery of pits. However, some groups of MFs grow into the pits but then terminate at the edge of the thickening, indicating that cellulose-synthase complexes are inactivated in the plasma membrane under the pit. Microtubules of TEs of both Pisum and Commelina are localized under the secondary thickening and few MTs are detected in the areas between wall thickenings. In the presence of the MT-disrupting agent, amiprophosmethyl, cellulose and hemicellulose, which is specific to secondary thickening, are deposited in deformed patterns in TEs of Pisum roots, Pisum epicotyls and Commelina roots. This indicates that the localized deposition of hemicellulose as well as cellulose involves MTs. The deformed, but heterogeneous pattern of secondary thickening is still visible, indicating that MTs are involved in determining and maintaining the regular patterns of the secondary thickening but not the spatial heterogeneous pattern of the wall deposition. A working hypothesis for the formation of the secondary thickening is proposed.Abbreviations APM amiprophosmethyl - DMSO dimethyl sulfoxide - F-WGA fluorescein-conjugated wheat-germ agglutinin - M F microfibril - MT microtubule - PEG polyethyleneglycol - TE tracheary element I thank Ms. Aiko Hirata (Institute of Applied Microbiology, University of Tokyo, Japan) for help in taking stereomicrographs. This work was supported in part by a Grant-in-Aid from the Ministry of Education, Science and Culture of Japan.     

Application of an enzyme-linked immunosorbent assay for screening of T-2 toxin-producing Fusarium spp.

Culture filtrates of Fusarium species were subjected without clean-up procedures to an indirect competitive enzyme-linked immunosorbent assay with anti-T-2 toxin monoclonal antibody. Fusarium sporotrichioides, F. poae, F. tricinctum, and F. culmorum strains were positive for T-2 toxin, with a minimum detection limit of 5 pg per assay (100 pg/ml of culture filtrate), and the assay data correlated well with the gas-liquid chromatographic data.     

Specific receptors for thromboxane A2 in cultured vascular smooth muscle cells of rat aorta

The specific binding site for thromboxane A2 (TXA2) was studied in cultured vascular smooth muscle cells (VSMC) of the rat aorta. [3H]SQ29,548, a potent and selective TXA2 receptor antagonist, displayed high-affinity and specificity, as well as saturable and displaceable binding to rat VSMC in culture. Scatchard analysis of equilibrium binding at 24 degrees C revealed a single class of binding sites with a Kd of 1.7 nM and a Bmax of 8.0 fmol/10(6) cells. A series of TXA2 receptor antagonists completely suppressed [3H]SQ29,548 binding to rat VSMC, and the rank order of their inhibitory potencies (Ki) correlated well with the potencies for suppression of the U46619-induced contraction of rat thoracic aorta. These results suggest that specific binding sites for [3H]SQ29,548 represent the TXA2 receptor in rat VSMC.     

Branching Photocycle of Sensory Rhodopsin in Halobacterium Halobium

Temperature effect of the photocyle of sensory rhodopsin (sR) was studied by nanosecond spectroscopy. Though the formation yield of sR_M (sR₃₇₀) was sharply decreased with temperature, those of sR_K (sR₆₈₀) and sR_L were insensitive to temperature changes. These results show the existence of the branching process back to sR from sR_L. The absorption maxima for sR_K and sR_L were 595 ± 5 and 555 ± 15 nm, respectively.     

Immunofluorescence Microscopy of Microtubule Arrangement in Root Cells of Pisum sativum L. var Alaska

The arrangement of wall microtubules (MTs) in Pisum sativumroots was viewed immunofluorescently using cryosectioning. Mostcells in the tip region of pea roots (0–2 mm from tip)had wall MTs arranged transversely to the root axis. In theregion elongating at a higher rate (2–4 mm), wall MTsof epidermal, cortical and stelar cells were all transverselyarranged. In the region of about 5 mm from the tip, in whichcell elongation had already ceased, wall MTs in cortical cellschanged from a transverse to an oblique arrangement in relationto the root axis. Some cells had a crossed arrangement of wallMTs, which was interpreted as representing two sets of unidirectional,oblique wall MTs in opposite cell cortices of a single cell.This change was completed within a region of 1-mm width. Sinceroots elongated at a rate of 0.6 mm h^–1, it means thatthe arrangement of wall MTs changed within 2 h. An oblique arrangementof wall MTs was also observed in stelar cells. As the cellsaged, the oblique arrangement tended to change to a steeperor even a longitudinal one. (Received January 24, 1986; Accepted May 15, 1986)     

An investigation for the occurrence of C4 photosynthesis in the Cyperaceae from Australia

Two hundred and twenty species of 38 genera in the Cyperaceae from Australia were examined for the possible occurrence of the C₄ photosynthesis and the anatomical features of leaves and culms. The Kranz type of anatomy and the carbon isotope ratios typical of C₄ plants were found in 84 species in the following six genera of four tribes belonging to subfamily Cyperoideae:Bulbostylis, Crosslandia, andFimbristylis (Fimbristylideae);Lipocarpha (Lipocarpheae);Cyperus (Cypereae);Rhynchospora (Rhynchosporeae). The anatomical observation revealed that the C₄ species possessed any one of the three Kranz anatomical types found by previous investigators. It was suggested that in the Cyperaceae the C₄ syndrome evolved independently within several taxa of the subfamily. The relative distribution of C₃ and C₄ species of the Cyperaceae in Australia was investigated by use of floristic data. It was recognized that the C₄ species dominated in the northern part of the continent which was characterized by tropical and subtropical savannas and hot dry areas with summer rainfall, and the C₃ species in the southern part, which contained temperate areas and mediterranean climatic areas with winter rainfall.     

Variations of Natural 13C Abundances in Leguminous Plants

The natural ¹³C abundance (¹³C value) of the field-grown leguminousplants (soybean, kidney bean, pea, azuki bean, mung bean, peanutand cowpea) was investigated by mass spectrometry with a precisionbetter than %0.2 for ¹³C. Among organs of premature plants,the leaves had the most negative values, and the nodules generallyhad the least negative values, and other organs, fruits, stemsand roots, showed intermediate values. In the soybeans so farinvestigated, the grains of nodulating plants exhibited higher¹³C values than nonnodulating lines. The ¹³C values of the grainsvaried depending on the species: peanuts showed the most negativevalues. Possible causes underlying these variations are discussed. ³Institute of Biological Sciences, University of Tsukuba, Sakura-mura,Ibaraki 305, Japan. (Received December 3, 1982; Accepted May 25, 1983)