Compartmentation of Aldolase Isoforms in Maize Leaves

The leaves of maize seedlings contain two principal isozymesof fructose 1,6-bisphosphate aldolase (E.C. 4.1.2.13 [EC] ), one chloroplasticand one cytosolic (Gasperini and Pupillo, 1982). Mesophyll protoplastswere separated from bundle sheath (BS) strands of both light-grownand dark-grown maize leaves. Aldolase isozymes were separatedfrom extracts of chloroplasts, etioplasts, protoplasts and BSstrands by column isoelectric focusing. The major isozyme ofgreen leaves (pI 4.2) was exclusively in BS chloroplasts, andthere was no evidence of other isozymes occurring in BS tissue.The cytosolic isozyme (pI 6.7) was present in protoplasts ofmesophyll cells, where it may limit the synthesis of hexose-phosphates(estimated activity of 9.4 µmol h^–1 g^–1 fr.wt.) together with lower activities of an acidic form (pI 4.6).Etiolated leaves contained significant amounts of the pI 6.7isozyme in both mesophyll and BS cells, but also minor activitiesof one or more acidic forms with pI values of 4.4–4.7(average pI 4.6) which appear to be located partly in BS etioplasts.The main developmental events for maize leaf aldolase afterillumination were a moderate decrease of cytosolic isozyme (pI6.7) which disappears from the BS within hours and a large,gradual increase of the BS plastid isozyme (pI 4.2). The isoformwith a pI 4.6 also increased rapidly to a low, steady activityin greening mesophyll protoplasts. Key words: C₄, fructose 1,6-bisphosphate, aldolase, Zea mays     

SPECIFIC ANOSMIAS TO 5{alpha}-ANDROST-16-EN-3-ONE AND {omega}-PENTADECALACTONE: THE URINOUS AND MUSKY PRIMARY ODORS

About 46% of human subjects are specifically anosmic to theodor of 5-androst-16-en-3- one, and about 9% are specificallyanosmic to the odor of -pentadecalactone. Odor threshold measurementson 33 representative steroids, synthetic musks and related compoundswere made with panels of normal observers and each of thesevarieties of specific anosmics. The androstenone anosmia wasmost pronounced with steroid ketones in the androstane seriesand some isosteric analogs. It is suggested that this specificanosmia corresponds with the absence of a recently-suspectedolfactory primary, the ‘urinous’ odor. The pentadecalactoneanosmia was associated with a number of diverse but approximatelyisosteric synthetic musks. This specific anosmia delineatesmore clearly the boundaries of the ‘musky’ odor,whose probably primacy has long been recognized. Threshold measurementswere made on mixtures of androst-16-en-3-one and pentadecalactone.The results depart little from the ‘rule of additivity’for odor ratios up to 100:1, and suggest a quantitative interpretationfor the anosmic defects measured with single compounds. ^* Permanent address: Istituto di Industrie Agrarie della Universitàdegli Studi, Pisa, Italy.     

Lower Cretaceous (Albian) shell-armoured and associated echinoid trace fossils from the Sácaras Formation, Serra Gelada area, southeast Spain

The Sácaras Formation (Albian, Lower Cretaceous) of the Serra Gelada succession (Prebetic of Alicante), southeast Spain, comprises carbonate‐rich, upwards thickening parasequences in which many types of trace fossils have been identified. The present study focuses on two types of tubular trace fossil characterized by features of their external coating. The first type is represented by a shell‐covered, structured trace fossil, up to 4?cm in diameter and 40?cm in length, built horizontally, from rectilinear (type 1) to gently curved (type 2), which envelopes an unstructured pipe of grey silty sediment. The coating is characterized by imbricated, flat particles, mainly orbitolinid foraminifers and other planar bioclasts, forming thin concentric layers; in cross section the bioclasts produce a typical plumed structure. This trace fossil represents a new ichnogenus and ichnospecies, here named Ereipichnus geladensis. Particle arrangement of the external coating is similar to that of terebelloid tubes, but Ereipichnus is a horizontal trace fossil, whereas structured worm tubes are vertical. The second type is a grain‐coated trace fossil, tubular in shape, with a simple internal structure. The coating is often reddish with respect to the neighbouring dark grey sediment and shows a slightly coarser‐grained texture, which envelopes the internal muddy pipe. This type, which yielded echinoids, was produced by irregular or heart‐shaped sea‐urchins (spatangoids) and is attributed to Scolicia or Cardioichnus. Facies analysis of the Serra Gelada succession with Ereipichnus and Scolicia or Cardioichnus locally shows other types of branched trace fossils (primarily represented by different forms of Thalassinoides) and bioturbation is developed in tiers, increasing upwards in abundance and diversity.     

Conservation of Scops Owl Otus scops in the Alps: relationships with grassland management, predation risk and wider biodiversity

The Scops Owl Otus scops is the most endangered and least studied owl in Europe, with widespread declines reported throughout Europe. The species is qualitatively associated with two threats: changes in agricultural practices and predation by Tawny Owls Strix aluco . We investigated these two threats to a population in the Alps, where land abandonment is causing widespread woodland expansion with unknown consequences. Predation risk and environmental indicators of the degree of agricultural change both predicted Scops Owl distribution. Furthermore, agricultural change also affected broader biodiversity, as estimated by the richness and diversity of bird and diurnal butterfly species. This resulted in a strong association between Scops Owl presence and wider biodiversity, which held at different spatial scales and justified a conservation focus on a single, threatened species. The persistence of Scops Owls in the Alps could be promoted through subsidies halting land abandonment and promoting extensive grassland management practices. Such interventions would have the additional advantage of yielding broader biodiversity benefits.     

Isolation of the Flagellar Swelling and Identification of Retinal in the Phototactic Flagellate, Ochromonas danica (Chrysophyceae)

Ochromonas danica, a freshwater, planktonic chrysophyte, is capable of sensing the light conditions of its environment. This biflagellate alga has a swelling near the base of the short flagellum and a chloroplastidic stigma in close association with it. A procedure is described for the isolation of this three dimensional flagellar swelling, the presumed photoreceptor. In contrast to an earlier method developed for the isolation of the paraflagellar swelling from Euglena gracilis, the protocol reported here for Ochromonas results in higher yields that should facilitate future biochemical investigations and could open avenues of investigation for the isolation and purification of the presumptive receptor protein. To verify the hypothesis that a rhodopsin-like protein might be present in this alga, we applied a standard extraction procedure successfully used in the identification of retinal. We here report the purification and identification of all-trans retinal in Ochromonas cells by column chromatography, HPLC and GC-MS. Since retinal is the chromophore of rhodopsin-like proteins, this finding may suggest that in these unicellular algae, too, a rhodopsin-like protein could be the photoreceptor pigment.     

Molecular phylogeny of Nitidulidae: assessment of subfamilial and tribal classification and formalization of the family Cybocephalidae (Coleoptera: Cucujoidea)

We present a molecular phylogeny of Nitidulidae based on thirty ingroup taxa representing eight of the ten currently recognized subfamilies. Approximately 10 K base pairs from seven loci (12S, 16S, 18S, 28S, COI, COII and H3) were used for the phylogenetic reconstruction. The phylogeny supports the following main conclusions: (i) Cybocephalidae are formally recognized as a distinct family not closely related to Nitidulidae and its constituent taxa are defined; (ii) Kateretidae are sister to Nitidulidae; (iii) Cryptarchinae are monophyletic and sister to the remaining nitidulid subfamilies; (iv) subfamily Prometopinae stat. res. is reinstated and defined, to accommodate taxa allied to Axyra Erichson, Prometopia Erichson and Megauchenia MacLeay; (v) Amphicrossinae, Carpophilinae and Epuraeinae are shown to be closely related taxa within a well‐supported monophyletic clade; (vi) tribal affinities and respective monophyly within Nitidulinae are poorly resolved by our data and must be more rigorously tested as there was little or no support for prior morphologically based tribes or genus‐level complexes; (vii) Nitidulinae are found to be paraphyletic with respect to Cillaeinae and Meligethinae, suggesting that they should either be subsumed as tribes, or Nitidulinae should be divided into several subfamilies to preserve the status of Cillaeinae and Meligethinae; (viii) Teichostethus Sharp stat. res. is not a synonym of Hebascus Erichson and the former is reinstated as a valid genus. These conclusions and emendations are discussed in detail and presented within a morphological framework.     

Genetic and environmental influences on shape variation in the European sea bass (Dicentrarchus labrax)

This paper demonstrates the contribution of both genetic and environmental effects on cultured European sea bass shape. We used the progeny of five populations of sea bass, in a partly diallel design, to investigate the genetics of shape (estimated with geometric morphometrics) in European sea bass. This was done using a common garden experiment with microsatellite markers assignment to parents and populations to avoid confusion between genetic and environmental effects. Additionally, one of the populations was studied over four different aquaculture facilities to investigate the effects of environment on shape. For the first time in this species, shape‐related traits were linked with genetic variation. The first relative warp analysis axis clearly differentiated rearing sites, demonstrating that the main shape/weight effects are related to culturing conditions, thereby accounting for ecomorphologically related differences. The second axis strongly differentiated groups by parental origins; there was a good correlation between shape differences and geographic distances between broodstock sampling locations. High heritabilities of axes scores (0.40–0.55) showed high genetic variation for shape within populations. This study shows that variation in shape has a high genetic component in sea bass, both at the population level and within populations. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 101 , 427–436.     

Isoforms of NADP-dependent Malic Enzyme in Tissues of the Greening Maize Leaf

Scagliarini, S., Pupillo, P. and Valenti, V. 1988. Isoformsof NADP-dependent malic enzyme in tissues of the greening maizeleaf.—J. exp. Bot. 39: 1109–1119. The compartmentation of the isoforms of NADP-dependent malicenzyme (E.C. 1.1.1.40 [EC] ) has been studied in cell-free extractsand in enzymatically-isolated protoplasts of mesophyll tissue(MT) and bundle sheath (BS) strands of greening maize leaves.The etiolated leaf of 10-d-old seedlings contains a cytosolicisozyme with a pl of 5.4 ?0.1 at low specific activity (s.a,45 ? 3 nmol min^–1 mg^–1 protein), found both in MTand BS. The green leaf on the other hand contains the dominantBS chloroplast isozyme with pl 4.6 ? 0.2 at a s.a, of 370 ?40 nmol min^–1 mg^–1 protein (3.2 ? 0.5 µmolmin^–1 mg^–1 chl) and a minor, previously undescribedisoform with pl 6.5 ? 0.1 also localized in the BS at a s.a.of 38 ? 6 nmol min^–1 mg^–1 protein. Green MT protoplastshave only traces of pl 4.6 isozyme. After illumination of dark-grown seedlings, the total leaf activityshows a rapid increase (1.5-fold within 2 h), attributed mainlyto the pl 5.4 isozyme of MT protoplasts and BS strands. Thisis followed by a large increase of enzyme activity due to thecontinued rise of pl 5.4 isozyme for about 24 h and, after aninitial lag of a few hours, to the accumulation of pl 4.6 isozyme.After 18 h illumination, pl 4.6 and 5.4 isozyme activities tendto decline in the MT whereas they are still increasing in theBS, particularly the former. This pl 4.6 species has becomethe major one by 48 h illumination. The final pattern of greenleaves is established around 96 h light, when the chloroplastisozyme has attained its maximum level, the pl 5.4 isozyme ofBS strands has been superceded by the pl 6.5 species (also supposedto be cytosolic) and MT protoplasts retain little residual activity.Some metabolic implications of the changing pattern of NADP-dependentmalic isozymes during maize leaf greening are discussed. Key words: C₄, isozymes, malic enzyme, photodifferentiation, Zea mays     

The Distribution of some NADP-Linked Dehydrogenases in Photosynthetic Tissues5

Mesophyll protoplasts of one-month-old maize leaves were separatedenzymatically from bundle sheath strands, and purified by centrifugationthrough a Percoll layer. The protoplasts and BS strands wereessentially pure as judged by microscopy, chl a/b ratios, andlevels of enzyme markers (PEP carboxylase and NADP-malic enzyme).Chioroplasts were obtained from the protoplasts and from homogenates,and purified through Percoll. The distribution of four NAD P-linked dehydrogenases in tissuesand organdies was examined. NADP-triose phosphate dehydrogenasc,used as a chloroplast marker, shows high and comparable specificactivities in both main tissues. Glucose 6-phosphate dehydrogenaseis located mainly in the mesophyll (at a specific activity of15.1 µmol h^–1 mg^–1chl in protoplasts) andis exclusively cytosolic. 6-Phosphogluconate dehydrogenase,also present in both tissue types, has a higher activity inthe BS (12.6 in purified strands versus 7.3 µmol h^–1mg^–1 chl in protoplasts). It is a cytosolic enzyme, althoughplastids may contain a low activity. Glyceraldehyde 3-phosphate:NADP reductasc is entirely in the mcsophyll cytoplasm (11.2µmol h^–1 mg^–1 chl). It is suggested that thecytoplasm of mcsophyll cells is a site of diversion of sugarphosphates for production of NADPH, at rates, however, compatiblewith the operation of the triose phosphate shuttle to bundlesheath cells for the synthesis of starch. Key words: Glucose-6-phosphate dehydrogenase, 6-phosphogluconate dchydrogenase, glyceralde-hyde-3-phosphate : NADP reductase, Zea mays