Ferredoxin–NADP+ Reductase from Tsuga canadensis. A Procedure for Isolation and Properties

Activity of ferredoxin-NADP⁺ reductase in leaf extracts of eastern hemlock [Tsuga canadensis (L.) Carr.] was relatively low, but could be markedly increased by use of protective agents. The best method employed polyvinylpolypyrrolidone (PVP) in the extraction medium plus removal of phenolic compounds by filtering the extracts through an insoluble PVP (Polyclar AT) column. Further purification of the enzyme was achieved by means of DEAE cellulose chromatography and DEAE Sephadex chromatography. A 94-fold purification of the enzyme with a total recovery of 43% was obtained. The eastern hemlock ferredoxin-NADP⁺ reductase was characterized by its diaphorase activity, i.e. the transfer of electrons from NADPH to an electron acceptor. 2,6-dichlorophenol indophenol. The pH optimum for the oxidation of NADPH is between 8.5 and 9.0. The enzyme is highly specific for its electron donor. NADPH, but shows low specificity for electron acceptors. The apparent Michaelis constant values of the enzyme for NADPH. NADH, and 2,6-dichlorophenol indophenol are 2.4 × 10^?5, 5.4 × 10^?3, and 4.7 × 10^?5M respectively. The molecular weight of the enzyme, as estimated by gel filtration, is about 45,000. The enzyme is inhibited by both organic and inorganic mercurials and certain cations. Comparison of properties of eastern hemlock ferredoxin-NADP⁺ reductase and spinach ferredoxin-NADP⁺ reductase shows that both enzymes are similar.     

RNA Synthesis in Division Synchronized Tetrahymena: Macronuclear and Cytoplasmic RNA*

SYNOPSIS. Synthesis of RNA in the macronucleus and appearance of RNA in the cytoplasm were studied in heat synchronized Tetrahymena pyriformis GL and compared to those found under conditions of logarithmic growth (28 C) and during heat shocks (34 C). In macronuclei of logarithmically growing cells precursors were processed to 2 rRNA species (25S and 17S). In addition, another RNA (15S), more homogeneous than the RNA (8-15S) in the cytoplasm, was observed in the macronucleus. Both 17S and 25S rRNA species were found in the cytoplasm, 17S rRNA appearing more rapidly than 25S rRNA. Synthesis of rRNA was suppressed at 34 C in cells subjected to heat synchronization; 8-15S RNA synthesis appeared to be inhibited to a lesser extent. During the time preceding the first synchronized division, the synthesis of rRNAs in the macronucleus slowly recovered. Early in the cycle, almost no newly synthesized rRNAs were extracted. By 30 min after the last heat shock (EH), most of the RNA synthesized was not identified as rRNA. By 60 min after EH, the pattern of RNA synthesis had not returned to that observed in logarithmically growing cells.     

The taxonomy of some British Littorina species, with notes on their reproduction (Mollusca: Prosobranchia)

The rough periwinkle Littorina saxatilis is considered, in current literature, to consist of various sympatric "subspecies" and "varieties". These, it has been suggested, are incipient species and are evidence for sympatric speciation.
This paper shows that in Wales the "saxatilis" aggregate consists of four separate, fully sympatric species: L. rudis, patula, nigrolineata and neglecta . Differences between these species include proportions, sculpture and size of the shell, range of colour patterns, the anatomy of the genitalia, and isozyme patterns. The possible evidence for incipient species is discussed and rejected.
The diversity of penis form amongst the British winkles is remarkable, and it is suggested that in these sympatric species the structure of the penis could be of importance in species recognition.
The four species occupy different zones on the shore and three certainly differ also in their methods of reproduction: L. rudis is ovoviviparous and contains embryos throughout the year, patula is ovoviviparous but contains embryos only in winter, nigrolineata is oviparous. The reproduction and dispersal methods of all British winkles are reviewed. The evidence available supports the view that non-planktonic dispersal is an adaptation which, in species whose shell colour varies considerably from one locality to another, enables each population to become adapted to its local environmental conditions. Ovoviviparity occurs in high-shore non-planktonic species, and may be an adaptation to avoid the dangers of egg desiccation. Planktonic dispersal is found in species whose shell colour does not vary as much as in non-planktonic ones. In these species, it is argued that a common planktonic pool enables a rapid re-colonization in the case of local decrease in population numbers, and also the rapid dispersal of successful mutations.     

A LuxR homologue of Xanthomonas oryzae pv. oryzae is required for optimal rice virulence

In Gram-negative bacteria a typical quorum sensing (QS) system usually involves the production and response to acylated homoserine lactones (AHLs). An AHL QS system is most commonly mediated by a LuxI family AHL synthase and a LuxR family AHL response regulator. This study reports for the first time the presence of a LuxR family-type regulator in Xanthomonas oryzae pv. oryzae ( Xoo ), which has been designated as OryR. The primary structure of OryR contains the typical signature domains of AHL QS LuxR family response regulators: an AHL-binding and a HTH DNA binding motif. The oryR gene is conserved among 26 Xoo strains and is also present in the genomes of close relatives X. campestris pv. campestris and X. axonopodis pv. citri . Disrupting oryR in three Xoo strains resulted in a significant reduction of rice virulence. The wild-type Xoo strains do not seem to produce AHLs and analysis of the Xoo sequenced genomes did not reveal the presence of a LuxI-family AHL synthase. The OryR protein was shown to be induced by macerated rice and affected the production of two secreted proteins: a cell-wall-degrading cellobiosidase and a 20-kDa protein of unknown function. By expressing and purifying OryR it was then observed that it was solubilized when grown in the presence of rice extract indicating that there could be a molecule(s) in rice which binds OryR. The role of OryR as a possible in planta induced LuxR family regulator is discussed.     

Integrative taxonomy,or iterative taxonomy?

The recently introduced term ‘integrative taxonomy’ refers to taxonomy that integrates all available data sources to frame species limits. We survey current taxonomic methods available to delimit species that integrate a variety of data, including molecular and morphological characters. A literature review of empirical studies using the term ‘integrative taxonomy’ assessed the kinds of data being used to frame species limits, and methods of integration. Almost all studies are qualitative and comparative – we are a long way from a repeatable, quantitative method of truly ‘integrative taxonomy’. The usual methods for integrating data in phylogenetic and population genetic paradigms are not appropriate for integrative taxonomy, either because of the diverse range of data used or because of the special challenges that arise when working at the species/population boundary. We identify two challenges that, if met, will facilitate the development of a more complete toolkit and a more robust research programme in integrative taxonomy using species tree approaches. We propose the term ‘iterative taxonomy’ for current practice that treats species boundaries as hypotheses to be tested with new evidence. A search for biological or evolutionary explanations for discordant evidence can be used to distinguish between competing species boundary hypotheses. We identify two recent empirical examples that use the process of iterative taxonomy.     

Sandhill Crane Abundance and Nesting Ecology at Grays Lake,Idaho

Abstract: We examined population size and factors influencing nest survival of greater sandhill cranes (Grus canadensis tabida) at Grays Lake National Wildlife Refuge, Idaho, USA, during 1997–2000. Average local population of cranes from late April to early May, 1998–2000, was 735 cranes, 34% higher than that reported for May 1970–1971. We estimated 228 (SE = 30) nests in the basin core (excluding renests), 14% higher than a 1971 estimate. Apparent nest success in our study (x̄ = 60%, n = 519 nests) was lower than reported for Grays Lake 30–50 years earlier. Daily survival rates (DSRs) of all nests averaged 0.9707 (41.2%). The best model explaining nest survival included year and water depth and their interaction. Nest survival was highest (DSR = 0.9827) in 1998 compared with other years (0.9698-0.9707). Nest survival changed little relative to water depth in 1998, when flooding was extensive and alternative prey (microtines) irrupted, but declined markedly with lower water levels in 2000, the driest year studied. Hypotheses relating nest survival to vegetation height, land use (idle, summer grazing, fall grazing), and date were not supported. In a before-after-control-impact design using 12 experimental fields, nest survival differed among years but not among management treatments (idle, fall graze, fall burn, and summer—graze—idle rotation), nor was there an interaction between year and treatments. However, DSRs in fall-burn fields declined from 0.9781 in 1997–1998 to 0.9503 in 1999–2000 (posttreatment). Changes in the predator community have likely contributed to declines in nest success since the 1950s and 1970s. Our results did not support earlier concerns about effects of habitat management practices on crane productivity. Nest survival could best be enhanced by managing spring water levels. Managers should continue censuses during late April to evaluate long-term relationships to habitat conditions and management.     

OXIDATION OF PHLORIDZIN BY ISOLATED CHLOROPLASTS

Phloridzin was shown to be oxidized by chloroplast fragmentsfrom swiss-chard. From inhibitor studies, kinetics and affinitytoward oxygen, it was inferred that the oxidation was mediatedby a phenolase in a "cresolase" type reaction. Atebrin was foundto inhibit the enzymatic oxidation of phloridzin and of 4-methylcatechol. (Received November 2, 1966; )     

Potato virus X: structure, disassembly and reconstitution

This paper summarizes some structural characteristics of Potato virus X (PVX), the flexuous filamentous plant potexvirus. A model of PVX coat protein (CP) tertiary structure in the virion proposed on the basis of tritium planigraphy combined with predictions of the protein tertiary structure is described. A possible role of glycosylation and phosphorylation in the CP structure and function is discussed. Two forms of PVX virion disassembly are discussed: (i) the virion co-translational disassembly after PVX CP in situ phosphorylation and (ii) disassembly of PVX triggered by different factors after linear destabilization of the virion by binding of the PVX-coded movement protein (TGBp1) to one end of the polar CP-helix. Special emphasis was placed on a translational activation of encapsidated PVX RNA and rapid disassembly of TGBp1-PVX complexes into free RNA and CP. The results of experiments on the PVX CP repolymerization and PVX reconstitution are considered. In particular, the products assembled from PVX RNA, CP and TGBp1 were examined. Single-tailed particles were found with a helical, head-like structure consisting of helically arranged CP subunits located at the 5'-tail of RNA; the TGBp1 was bound to the end of the head. Translatable 'RNA-CP-TGBp1' complexes may represent the transport form of the PVX infection.     

Climate controls on C3 vs. C4 productivity in North American grasslands from carbon isotope composition of soil organic matter

We analyzed the δ¹³C of soil organic matter (SOM) and fine roots from 55 native grassland sites widely distributed across the US and Canadian Great Plains to examine the relative production of C₃ vs. C₄ plants (hereafter %C₄) at the continental scale. Our climate vs. %C₄ results agreed well with North American field studies on %C₄, but showed bias with respect to %C₄ from a US vegetation database (statsgo ) and weak agreement with a physiologically based prediction that depends on crossover temperature. Although monthly average temperatures have been used in many studies to predict %C₄, our analysis shows that high temperatures are better predictors of %C₄. In particular, we found that July climate (average of daily high temperature and month's total rainfall) predicted %C₄ better than other months, seasons or annual averages, suggesting that the outcome of competition between C₃ and C₄ plants in North American grasslands was particularly sensitive to climate during this narrow window of time. Root δ¹³C increased about 1‰ between the A and B horizon, suggesting that C₄ roots become relatively more common than C₃ roots with depth. These differences in depth distribution likely contribute to the isotopic enrichment with depth in SOM where both C₃ and C₄ grasses are present.     

Indirect effects of introduced trout on Cascades frogs (Rana cascadae) via shared aquatic prey

1. The introduction of trout to montane lakes has negatively affected amphibian populations across the western United States. In northern California’s Klamath–Siskiyou Mountains, introduced trout have diminished the distribution and abundance of a native ranid frog, Rana (=Lithobates) cascadae. This is primarily thought to be the result of predation on frog larvae. However, if trout feed on larval aquatic insects that are available to R. cascadae only after emergence, then resource competition may also affect this declining native amphibian. 2. Stomach contents of R. cascadae were compared between lakes that contained trout and those from which introduced trout were removed. Total prey mass in stomach contents relative to frog body mass was not significantly different between lakes with fish and fish‐removal lakes, but in the former R. cascadae consumed a smaller proportion of adult aquatic insects. The stomach contents of fish included larvae of aquatic insects that are, as adults, eaten by R. cascadae. 3. Rana cascadae consumed fewer caddisflies (Trichoptera) and more grasshoppers (Orthoptera) at lakes with higher densities of fish. At lakes with greater aquatic habitat complexity, R. cascadae consumed more water striders (Hemiptera: Gerridae) and terrestrial spiders (Araneae). 4. We suggest that reductions in the availability of emerging aquatic insects cause R. cascadae to consume more terrestrial prey where trout are present. Thus, introduced trout may influence native amphibians directly through predation and, indirectly, through pre‐emptive resource competition.