Gametic differentiation in Chlamydomonas reinhardtii. I. Production of gametes and their fine structure

Gametogenesis in Chlamydomonas reinhardtii has been studied in mating-type plus cells utilizing several different culture conditions, all of which are shown to depend on the depletion of nitrogen from the medium, and the fine structure of gametes prepared under these conditions has been compared by using thin sections of fixed materials. We document alterations in ribosome levels, in chromatin morphology, in starch levels, in the organization of chloroplast membranes, and in the appearance of nuclear envelope and endoplasmic reticulum membranes during gametogenesis. We also noted the acquisition of two new organelles: a mating structure (Friedman, L., A. L. Colwin, and L. H. Colwin. 1968. j. cell Sci. 3:115-128; goodenough, U. W., and R. L. Weiss. 1975. J. Cell Biol. 67:623-637), and Golgi-derived vesicles containing a homogeneous material. We chart the time course of these morphological changes during synchronous gametogenesis. We note that many of these changes may represent adjustments to nitrogen starvation rather than direct features of gametic differentiation, and we also document that cells can differentiate so that they survive conditions of nitrogen starvation for many weeks after they become gametes. We conclude that metabolic alterations, the acquisition of mating ability, and the preparation for long-term survival are all elicited in this organism by nitrogen withdrawal, and we discuss how the various structural alterations observed in this study may relate to these three interrelated avenues of cellular differentiation.     

Crystals of the Chlamydomonas reinhardtii cell wall: polymerization, depolymerization, and purification of glycoprotein monomers

Two of the three major outer layers of the Chlamydomonas reinhardtii cell wall (W6 and W4) can be solubilized from living cells with sodium perchlorate or other chaotropes and will repolymerize in vitro to form milligram amounts of wall crystals. Conditions for optimal crystalization are presented, and conditions that fail to induce polymerization are exploited to maintain monomers in aqueous solution for ion-exchange chromatography. The four major glycoproteins of the complex (GP1, 1.5, 2, and 3) have in this way been purified to apparent homogeneity and have been characterized morphologically by transmission electron microscopy using the quick-freeze, deep-etch technique and by amino acid composition. Three of the four are hydroxyproline-rich species that copolymerize to form the W6 layer. The fourth (GP1.5) is a glycine-rich species that binds to the interior of the in vitro crystal; it is apparently equivalent to the granules within the W4 layer in situ.     

Alterations to the catalytic properties of polyphenoloxidase in detergentless microemulsions and ternary water-organic solvent mixtures

Summary The catalytic properties of mushroom polyphenoloxidase could be substantially altered by entrapment into hexane- and toluene-based microemulsions stabilized with isopropanol. The fast irreversible inactivation and drastic substrate inhibition of the enzyme were significantly reduced in detergentless microemulsions in comparison to conventional aqueous media. Similar changes in the catalytic behavior of polyphenoloxidase were observed in the normal ternary solutions of hexane-(toluene)-isopropanol-water, and in the H-bonded aggregates of isopropanol and water in toluene, but not in hexane.     

Quantifying the robustness of first arrival dates as a measure of avian migratory phenology

As the climate changes, many long‐term studies have shown that the timing of bird migration is shifting, increasing the need for reliable measures of migratory phenology. Ideally, daily counts of birds at a site are used to calculate the mean arrival date (MAD) but, as this approach is not always possible and is very labour‐intensive, simpler metrics such as first arrival date (FAD) have commonly been used. Here, we examine the relationship between FAD and MAD in 28 summer migrant bird species over a 42‐year period (1970–2011) at Portland Bird Observatory, UK. Although significant correlations between FAD and MAD were detected, relationships were weak, particularly in long‐distance migrants. We suggest that FAD, although a simple and straightforward measure, is not particularly robust as a proxy for overall migratory phenology at a population level.     

High-pressure liquid chromatography fractionation of Chlamydomonas dynein extracts and characterization of inner-arm dynein subunits

A rapid procedure for fractionating salt-stable dynein subunits from high-salt extracts of Chlamydomonas axonemes has been developed using a high-pressure liquid chromatography system with an anion exchange column and gradient salt elution. Five distinct fractions are shown to be highly enriched for five distinct subunits or subunit complexes by SDS/polyacrylamide gel electrophoresis. ATPase activity and electron microscopy. Peaks 1 and 4 contain, respectively, the single-headed gamma-subunit and the two-headed alpha/beta-heteropolymer that form the outer arm in situ and are dissociated by salt exposure; both peaks are absent from the outer arm-less mutant pf-28. Peaks 2, 3 and 5 contain, respectively, two distinct single-headed species and a double-headed species that derive from inner arms; all three peaks are missing from the inner arm-less mutant pf-23. Sucrose-gradient sedimentation analysis confirms these assignments and provides additional information on the intermediate-chain and light-chain composition of the inner-arm species. Electron microscopy of the purified inner-arm species visualized by the quick-freeze deep-etch technique complements a previous analysis of outer-arm species. Each protein is shown to have a unique morphology, and both the inner- and outer-arm proteins clearly belong to a common family whose structural divergence presumably reflects functional specialization.     

Can Handheld Thermal Imaging Technology Improve Detection of Poachers in African Bushveldt?

Illegal hunting (poaching) is a global threat to wildlife. Anti-poaching initiatives are making increasing use of technology, such as infrared thermography (IRT), to support traditional foot and vehicle patrols. To date, the effectiveness of IRT for poacher location has not been tested under field conditions, where thermal signatures are often complex. Here, we test the hypothesis that IRT will increase the distance over which a poacher hiding in African scrub bushveldt can be detected relative to a conventional flashlight. We also test whether any increase in effectiveness is related to the cost and complexity of the equipment by comparing comparatively expensive (22000 USD) and relatively inexpensive (2000 USD) IRT devices. To test these hypotheses we employ a controlled, fully randomised, double-blind procedure to find a poacher in nocturnal field conditions in African bushveldt. Each of our 27 volunteer observers walked three times along a pathway using one detection technology on each pass in randomised order. They searched a prescribed search area of bushveldt within which the target was hiding. Hiding locations were pre-determined, randomised, and changed with each pass. Distances of first detection and positive detection were noted. All technologies could be used to detect the target. Average first detection distance for flashlight was 37.3m, improving by 19.8m to 57.1m using LIRT and by a further 11.2m to 68.3m using HIRT. Although detection distances were significantly greater for both IRTs compared to flashlight, there was no significant difference between LIRT and HIRT. False detection rates were low and there was no significant association between technology and accuracy of detection. Although IRT technology should ideally be tested in the specific environment intended before significant investment is made, we conclude that IRT technology is promising for anti-poaching patrols and that for this purpose low cost IRT units are as effective as units ten times more expensive.     

What prevents phenological adjustment to climate change in migrant bird species? Evidence against the “arrival constraint” hypothesis

Phenological studies have demonstrated changes in the timing of seasonal events across multiple taxonomic groups as the climate warms. Some northern European migrant bird populations, however, show little or no significant change in breeding phenology, resulting in synchrony with key food sources becoming mismatched. This phenological inertia has often been ascribed to migration constraints (i.e. arrival date at breeding grounds preventing earlier laying). This has been based primarily on research in The Netherlands and Germany where time between arrival and breeding is short (often as few as 9 days). Here, we test the arrival constraint hypothesis over a 15-year period for a U.K. pied flycatcher (Ficedula hypoleuca) population where laying date is not constrained by arrival as the period between arrival and breeding is substantial and consistent (average 27 ± 4.57 days SD). Despite increasing spring temperatures and quantifiably stronger selection for early laying on the basis of number of offspring to fledge, we found no significant change in breeding phenology, in contrast with co-occurring resident blue tits (Cyanistes caeruleus). We discuss possible non-migratory constraints on phenological adjustment, including limitations on plasticity, genetic constraints and competition, as well as the possibility of counter-selection pressures relating to adult survival, longevity or future reproductive success. We propose that such factors need to be considered in conjunction with the arrival constraint hypothesis.     

A Composite Gel–Polymer/Glass–Fiber Electrolyte for Sodium‐Ion Batteries

An integrated preparation of a low‐cost composite gel–polymer/glass–fiber electrolyte with poly(vinylidene fluoride‐co‐hexafluoropropylene) (PVDF‐HFP) reinforced by a glass–fiber paper and modified by a polydopamine coating to tune the mechanical and surface properties of PVDF‐HFP is shown to be applicable to a sodium‐ion battery. The composite polymer matrix exhibits excellent mechanical strength and thermal stability up to 200 °C. After saturating with a liquid electrolyte, a wide electrochemical window and high ionic conductivity is obtained for the composite gel–polymer/glass–fiber electrolyte. When tested in a sodium‐ion battery with Na₂MnFe(CN)₆ as cathode, the rate capability, cycling performance, and coulombic efficiency are significantly improved. The results suggest that the composite polymer electrolyte is a very attractive separator for a large‐scale battery system where safety and cost are the main concerns.