The Value of Molecular vs. Morphometric and Acoustic Information for Species Identification Using Sympatric Molossid Bats

A fundamental condition for any work with free-ranging animals is correct species identification. However, in case of bats, information on local species assemblies is frequently limited especially in regions with high biodiversity such as the Neotropics. The bat genus Molossus is a typical example of this, with morphologically similar species often occurring in sympatry. We used a multi-method approach based on molecular, morphometric and acoustic information collected from 962 individuals of Molossus bondae, M. coibensis, and M. molossus captured in Panama. We distinguished M. bondae based on size and pelage coloration. We identified two robust species clusters composed of M. molossus and M. coibensis based on 18 microsatellite markers but also on a more stringently determined set of four markers. Phylogenetic reconstructions using the mitochondrial gene co1 (DNA barcode) were used to diagnose these microsatellite clusters as M. molossus and M. coibensis. To differentiate species, morphological information was only reliable when forearm length and body mass were combined in a linear discriminant function (95.9% correctly identified individuals). When looking in more detail at M. molossus and M. coibensis, only four out of 13 wing parameters were informative for species differentiation, with M. coibensis showing lower values for hand wing area and hand wing length and higher values for wing loading. Acoustic recordings after release required categorization of calls into types, yielding only two informative subsets: approach calls and two-toned search calls. Our data emphasizes the importance of combining morphological traits and independent genetic data to inform the best choice and combination of discriminatory information used in the field. Because parameters can vary geographically, the multi-method approach may need to be adjusted to local species assemblies and populations to be entirely informative.     

Molecularly cloned simian immunodeficiency virus SIVagm3 is highly divergent from other SIVagm isolates and is biologically active in vitro and in vivo.

Simian immunodeficiency viruses have been isolated from African green monkeys originating from Ethiopia. A molecular clone, termed SIVagm3, was found to be highly divergent from SIVagmTYO-1 in terms of its restriction map and partial nucleotide sequence. A premature stop codon present in the transmembrane protein of SIVagm TYO-1 was absent in SIVagm3. SIVagm3 was biologically active in vitro and in vivo and displayed characteristics reminiscent of the wild-type virus. Biological activity was demonstrated by seroconversion of juvenile African green monkeys and Macaca nemestrina after inoculation. In contrast to antibody reactivity mainly directed against env proteins in naturally infected African green monkeys. African green monkeys and M. nemestrina infected with the cloned virus showed antibody reactivity directed against all major proteins as demonstrated by immunoblot analysis. The availability of a biologically fully competent molecular clone of SIVagm allows us now to address various pertinent questions in an animal model system which should help to understand features of human immunodeficiency virus infection in human beings.     

Ultrastructural localization of the high molecular weight proteins associated with in vitro-assembled brain microtubules

Microtubules isolated from brain extracts by in vitro assembly (1, 19, 23) are composed principally of two tubulins and two high molecular weight proteins (microtubule-associated proteins [MAPS] 1 and 2) (2,5,7,20). Recently, it was demonstrated that in vitro-assembled brain microtubules (neurotubules) are coated with filaments (5, 7) which are similar to the filaments attached to neurotubules in situ (4, 15, 21, 24, 25), and it was suggested that the filaments are composed of the higher molecular weight MAPs (5, 7, 12). In this study, microtubules were assembled in the presence and absence of the MAPs, and thin sections of the microtubules were examined by electron microscopy. The results show that the filaments only occur on microtubules assembled in the presence of the MAPs and it is therefore concluded that the filaments are composed of the high molecular weight MAP's.     

Development and replenishment of the P-depletion zone around the primary root of maize during the vegetation period

The dynamics of the development and replenishment of P-depletion zones around the primary root of maize (Zea mays L. cv ‘Garbo’) was studied during a vegetation period (80 days) under greenhouse conditions in a loamy sand of low P-availability. A recently described freeze-cutting technique was used to determine radial diffusion of labelled phosphate to the primary root. The development of the depletion zone was biphasic. In the initial phase after two days of growth of the primary root in a soil layer labelled with³³P a minimum of isotopically exchangeable P (EP) was observed which had decreased to about 30% of its original amount at the root surface. At that time the corresponding P-concentration in the soil solution was calculated to be as low as 5×10⁻⁷ M. The depletion zone had already spread 0.4 mm from the root surface. During the second phase, between the 10th and 20th day of plant growth the concentration of EP at the root surface increased slowly but did not change markedly. However, the depletion zone continued to spread and after the 20th day of growth reached its maximal diameter (1.07 mm from the root surface) but remained completely within the root hair cyclinder; the single root hairs never exceeded 1.14 mm in length. The biphasic growth of the depletion zone was probably caused by proton extrusion of the root tip. Acidification of the soil solution from pH 5.8 to about 3.9 results in an about 3-fold rise of the concentration of desorbed phosphate and might also have activated acidophilic P-translocators of the root during the initial phase. Anion over cation uptake normally prevailing during the later stage of root development might resulted in a rise of the soil pH within the root hair zone. Consequently P-availability, as well as P-uptake capacity declined, but P-uptake by the seminal root still continued until the 20th day. Subsequently, the P-concentration within the depletion zone increased again while simultaneously its extent was reduced until it was almost completely replenished after 60 days indicating a loss of P-uptake capacity of the primary root. Within the root tissue³³P was accumulated to about twice the concentration of that in the undepleted soils. This accumulation corresponded to periods of high uptake due to the development of root laterals. In the root cortex a high P-content was observed during the first 30 days of growth. At the onset of the reproductive stage of the plant the P-content of the shoot and especially in the developing seeds rose considerably at the cost of phosphate stored in the root cortex. The accumulation of³³P in the root tissue indicated that nutrient gain was mainly achieved during the early stages of plant development and that P was temporarily stored to some extent within the root system.     

Characterization of a Tetrahymena thermophila mutant strain unable to develop normal thermotolerance.

For Tetrahymena thermophila cells to survive extended periods of time at 43 degrees C, they must continuously synthesize heat shock proteins. For its translational machinery to function at 43 degrees C, T. thermophila requires either prior nonlethal heat shock treatment or brief treatment with partially inhibiting doses of cycloheximide or emetine. We have identified and characterized a mutant strain of T. thermophila (MC-3) in which prior nonlethal heat shock does not prevent protein synthesis inactivation at 43 degrees C. In addition, treatment of MC-3 cells with either of the antibiotics that normally confer 43 degrees C thermoprotection on wild-type cells elicited no similar thermoprotective response in these cells. Despite these phenotypic characteristics, by other criteria MC-3 synthesized a normal, functional array of heat shock proteins at 40 degrees C, a nonlethal heat shock protein-inducing temperature. The mutation in MC-3 which prevents the thermostabilization of protein synthesis by nonlethal heat shock is, by genetic criteria, most likely the same one which prevents the induction of thermotolerance by drug treatments. We present evidence that this mutation may affect some ribosome-associated functions.     

Zur Morphologie,Systematik und Funktion der Lymphgefässe

Zusammenfassung Die Kombination der H₂O₂-Methode mit mikroskopischer Betrachtung läßt säckchenförmige Anfänge des Lymphsystems erkennen, die als Lymphkapillaren bezeichnet werden. Aus morphologischen und physiologischen Gründen wird jedoch vorgeschlagen, statt der Bezeichnung Lymphkapillaren die Benennung Lymphalveolen oder Lymphrezeptoren zu gebrauchen. Anatomische und physiologische Befunde sprechen weiterhin dafür, daß diffusible Stoffe die Blut- und Lymphkapillarwände in beiden Richtungen durchsetzen können, undiffusible Stoffe hingegen die Richtung Blutkapillare—Gewebe und Gewebe—Lymphkapillare einhalten müssen (Ventilwirkung). Dieser Richtungszwang für undiffusible Stoffe auf das Lymphgefäß zu erklärt, warum z.B. in der Darmwand entstehendes Neutralfett oder ins Gewebe eingedrungene Bakterien in der Regel zunächst nur in die Lymphgefäße mit ihren Sondereinrichtungen, den Lymphknoten, gelangen können. Diese Kreislaufsteuerung wird unter anderem durch das bisher wenig beachtete, aber bei den meisten Lymphgefäßdarstellungen auftretende Phänomen der aus schließlichen Lymphgefäßfüllung begründet. Ein Schema veranschaulicht die Kreislaufverhältnisse im intrakapillären Bereich, die vorwiegend passiv, durch morphologische Unterschiede zwischen Blut- und Lymphkapillaren bedingt sein dürften.Die abführenden Lymphgefäße zeigen in ihren Zwischenklappenabschnitten eine Tropfenform, die als Folge starken seitlichen Druckes auf die in ihnen eingeschlossene Lymphe gewertet wird. Auf die Problematik der Bezeichnung Saugadern für die peripheren Lymphgefäße wird hingewiesen. Besonderheiten in der Wandstruktur der abführenden Lymphgefäße werden kurz erörtert.Herrn Prof. Dr. Grau danke ich für Auskünfte und Ratschläge, weiterhin Herrn Oberveterinärrat Dr. Mayer, Herrn Veterinärrat Dr. Sauer sowie Herrn Dr. Walter, München, für die Anfertigung der photographischen Bilder.     

Crystallization of a complex of hemoglobin components II and III of the symbiont-harboring clam Lucina pectinata.

Diffraction data to 3.1 A resolution were collected on crystals of a complex of components II and III of the cytoplasmic hemoglobin of the symbiont-harboring clam Lucina pectinata. The crystal system is tetragonal, a = 76.3 A, c = 153.1 A and the space group is P42212. The asymmetric unit probably contains a dimer of the tetrameric complex.     

Uniconazole inhibits stress-induced ethylene in wheat and soybean seedlings

Previous studies have shown that uniconazole inhibits ethylene synthesis and protects plants from various stresses. The present research was conducted to delineate the mechanism of ethylene inhibition by uniconazole [(E)-(p-chlorophenyl)-4,4-dimethyl-2-(1,2,4-triazol-1-yl)-1-penten-3-ol]. Following heat stress of 48°C for 3 h, the shoots of the control wheat seedlings became desiccated, and the seedlings lost 23% of their fresh mass 8 h after stress. The control soybean seedlings had epinastic unifoliate leaves 5 h after foliar application (4.4 g.a.i./ha) of the herbicide triclopyr [(3,5,6-trichloro-2-pyridinyl)oxyacetic acid]. Soil drench applications of uniconazole, a potent member of the triazole family, reduced these symptoms associated with heat and herbicide stress in wheat (5.0 mg/L) and soybean (0.4 mg/L) seedlings, respectively. Basal ethylene production was inhibited 32 and 48% by uniconazole in the wheat and acotyledonous soybean seedlings, respectively. Following a 48°C heat stress, 1-aminocyclopropane-1-carboxylic acid (ACC) levels increased 40% in both the control and uniconazole-treated wheat seedlings. After triclopyr application, ACC levels increased 400% in both the control and uniconazoletreated soybean seedlings. The increased ACC levels, following stress, were accompanied by increased ethylene production from the control, but not from the uniconazole-treated wheat and acotyledonous soybean seedlings. Uniconazole treatment did not significantly change the basal or stress-induced N-malonyl-1-aminocyclopropane-1-carboxylic acid (MACC) levels compared to controls. These results suggest that uniconazole inhibits ethylene synthesis by interfering with the conversion of ACC to ethylene in wheat and acotyledonous soybean seedlings. Ethylene production and ACC conversion were not inhibited by uniconazole in excised soybean cotyledons. These results indicate that different ethylene-forming enzyme (EFE) systems operate in the soybean acotyledonous seedling and cotyledon, and the system in the former is inhibited by uniconazole.