A mutation in aspergillus nidulans that blocks the transition from interphase to prophase

In order to develop a method for obtaining mitotic synchrony in aspergillus nidulans, we have characterized previously isolated heat-sensitive nim mutations that block the nuclear division cycle in interphase at restrictive temperature. After 3.5 h at restrictive temperature the mitotic index of a strain carrying one of these mutations, nimA5, was 0, but when this strain was subsequently shifted from restrictive to permissive temperature the mitotic index increased rapidly, reaching a maximum of 78 percent after 7.5 min. When this strain was examined electron-microscopically, mitotic spindles were absent at restrictive temperature. From these data we conclude that at restrictive temperature nimA5 blocks the nuclear division cycle at a point immediately preceding the initiation of chromosomal condensation and mitotic microtubule assembly, and upon shifting to permissive control over the initiation of microtubule assembly and chromosomal condensation in vivo through a simple temperature shift and, consequently, nimA5 should be a powerful tool for studying these processes. Electron-microscopic examination of spindles of material synchronized in this manner reveals that spindle formation, although very rapid, is gradual in the sense that spindle microtubule numbers increase as spindle formation proceeds.     

The evolution of alternative reproductive strategies: fitness differential, heritability, and genetic correlation between the sexes

Paternity analyses using molecular markers have become standard in studies of mating systems, parentage, and kinship. In systems where individuals exhibit alternative mating strategies, molecular analyses have been productively used to estimate the reproductive success of each behavioral type and hence the fitness consequences to each individual. Here we review the fitness results in a system of five alternative mating strategies present in one population of side-blotched lizards (Uta stansburiana). Males in this population adopt one of three behavioral strategies that differ in their degree of territoriality and mate guarding. In contrast, females adopt one of two strategies that differ in offspring quantity and quality. We use paternity analyses to estimate the fitness of each morph, the heritability of reproductive strategy, and the correlation in strategy between the sexes and discuss the implications of our findings for the evolution and maintenance of reproductive polymorphism in this and other systems.     

Uptake of amino acids and peptides by developing barley embryos

Developing embryos of barley ( Hordeum vulgare L. cv. Bomi) detached 21–27 days after anthesis took up 1 mM [¹⁴C]-glutamine at pH 5 and 30°C at a rate of about 20 nmol embryo^−l h⁻¹ (5 μol g⁻¹h⁻¹). The uptake was inhibited by about 50% by di-nitrophenol and by about 80% by 300 m M unlabelled glutamine or alanine. The bulk of the uptake appeared, therefore, to be due to carrier-mediated active transport. The pH optimum of the uptake was 4.5. Leucine, proline, lysine, arginine and as-paragine were taken up at approximately similar rates as glutamine, and they also inhibited the uptake of glutamine. This, suggests that the uptake of glutamine was at least partly due to an unspecific carrier(s) also shared by other amino acids. The embryos also took up the dipepti.de glycykarcosine; the rate was about 6 nmol embryo⁻¹h⁻¹ (1.5 μol g⁻¹h⁻¹) (2 mM glycylsarcosine, pH 4.5, 30°C). The uptake was inhibited by about 70% by dinitrophenol or by 300 m M glycylglycine. This indicates that the bulk of the uptake was due to carrier-mediated active transport. The pH optimum of the uptake was about 4.5.
The rates of glutamine and glycylsarcosine uptake increased during the early and middle stages of embryo development (until day 28 after anthesis), but decreased towards the end of the maturation of the grain. These changes, as well as the relatively high activities, suggest that carrier-mediated active uptake of amino acids, and possibly also that of peptides, plays a role in the nutrition of the developing embryo.     

Heritable body size mediates apparent life-history trade-offs in a simultaneous hermaphrodite

Physiological trade-offs between life-history traits can constrain natural selection and maintain genetic variation in the face of selection, thereby shaping evolutionary trajectories. This study examines physiological trade-offs in simultaneously hermaphroditic banana slugs, Ariolimax dolichophallus. These slugs have high heritable variation in body size, which strongly predicts the number of clutches laid, hatching success and progeny growth rate. These fitness components were associated, but only when examined in correlation with body size. Body size mediated these apparent trade-offs in a continuum where small animals produced rapidly growing progeny, intermediate-sized animals laid many clutches and large animals had high hatching success. This study uses a novel statistical method in which the components of fitness are analysed in a mancova and related to a common covariate, body size, which has high heritability. The mancova reveals physiological trade-offs among the components of fitness that were previously masked by high variation in body size.     

Discrete genetic variation in mate choice and a condition-dependent preference function in the side-blotched lizard: implications for the formation and maintenance of coadapted gene complexes

Variation in female preference functions, both genotypic andphenotypic, has been largely ignored in the literature, despiteits implications to the maintenance of genetic variation inpopulations and the resolution of the "Lek paradox." Polymorphicpopulations, such as in the side-blotched lizard, provide idealstudy systems for its investigation, especially in the contextof incipient processes of sympatric speciation. Females of theside blotch lizard exist in 2 genetically distinct morphs, yellowthroated and orange throated, that experience disruptive selectionfor life history traits. Males express 3 throat color morphs,blue, orange, and yellow, that exhibit alternative strategiesin intrasexual competition. We experimentally tested for femalepreference in triadic mate choice trials to identify the presenceof discrete genetic and condition-dependent variation in femalepreference function. We found that females did in fact showgenetic variation in preference for males but that females alsooperate a multicondition preference function dependent uponthe genotype of the female and her state (number of clutcheslaid). Females exhibited positive assortative mating prior tothe first clutch. However, prior to later clutches, orange femalesswitched choice, preferring yellow males. These findings arediscussed in relation to the maintenance of coadapted gene complexeswithin populations and the prevention of divergent directionalselection (population bifurcation) by condition-dependent variationin mate choice.     

Direct fitness correlates and thermal consequences of facultative aggregation in a desert lizard

Social aggregation is a common behavioral phenomenon thought to evolve through adaptive benefits to group living. Comparing fitness differences between aggregated and solitary individuals in nature - necessary to infer an evolutionary benefit to living in groups - has proven difficult because communally-living species tend to be obligately social and behaviorally complex. However, these differences and the mechanisms driving them are critical to understanding how solitary individuals transition to group living, as well as how and why nascent social systems change over time. Here we demonstrate that facultative aggregation in a reptile (the Desert Night Lizard, Xantusia vigilis) confers direct reproductive success and survival advantages and that thermal benefits of winter huddling disproportionately benefit small juveniles, which can favor delayed dispersal of offspring and the formation of kin groups. Using climate projection models, however, we estimate that future aggregation in night lizards could decline more than 50% due to warmer temperatures. Our results support the theory that transitions to group living arise from direct benefits to social individuals and offer a clear mechanism for the origin of kin groups through juvenile philopatry. The temperature dependence of aggregation in this and other taxa suggests that environmental variation may be a powerful but underappreciated force in the rapid transition between social and solitary behavior.     

WHY DOES THE YELLOW‐EYED ENSATINA HAVE YELLOW EYES? BATESIAN MIMICRY OF PACIFIC NEWTS (GENUS TARICHA) BY THE SALAMANDER ENSATINA ESCHSCHOLTZII XANTHOPTICA

Color patterns commonly vary geographically within species, but it is rare that such variation corresponds with divergent antipredator strategies. The polymorphic salamander Ensatina eschscholtzii, however, may represent such a case. In this species, most subspecies are cryptically colored, whereas E. e. xanthoptica, the Yellow eyed ensatina, is hypothesized to be an aposematic mimic of highly toxic Pacific newts (genus Taricha). To test the mimicry hypothesis, we conducted feeding trials using Western Scrub-Jays, Aphelocoma californica. In every feeding trial, we found that jays, following presentation with the presumed model (T. torosa), were more hesitant to contact the presumed mimic (E. e. xanthoptica) than a control subspecies lacking the postulated aposematic colors (E. e. oregonensis). The median time to contact was 315 sec for the mimic and 52 sec for the control. These results support the mimicry hypothesis, and we suggest that E. e. xanthoptica is likely a Batesian mimic, rather a Müllerian or quasi-Batesian mimic, of Pacific newts.     

Corticosterone manipulation reveals differences in hierarchical organization of multidimensional reproductive trade-offs in r-strategist and K-strategist females

Life history trade-offs are often hierarchical with decisions at one level affecting lower level trade-offs. We investigated trade-off structure in female side-blotched lizards (Uta stansburiana), which exhibit two evolved strategies: yellow-throated females are K-strategists and orange-throated are r-strategists. Corticosterone treatment was predicted to differentially organize these females' reproductive decisions. Corticosterone-treated yellow females suppressed reproduction but survived well, and augmented egg mass without decreasing clutch size. Conversely, corticosterone enhanced mortality and reproductive rates in orange females, and increased egg mass only after lengthy exposure. Corticosterone did not affect post-laying condition, suggesting that corticosterone increased egg mass through enhanced energy acquisition (income breeding). Corticosterone enhanced survival of lightweight females, but decreased survival of heavy females, introducing a foraging vs. predation trade-off. We conclude that rather than being a direct, functional relationship, observed trade-offs between offspring size and number represent evolved differences in hierarchical organization of multidimensional trade-offs, particularly in response to stress.     

SOCIAL CAUSES OF CORRELATIONAL SELECTION AND THE RESOLUTION OF A HERITABLE THROAT COLOR POLYMORPHISM IN A LIZARD

Abstract When selection acts on social or behavioral traits, the fitness of an individual depends on the phenotypes of its competitors. Here, we describe methods and statistical inference for measuring natural selection in small social groups. We measured selection on throat color alleles that arises from microgeographic variation in allele frequency at natal sites of side‐blotched lizards (Uta stansburiana). Previous game‐theoretic analysis indicates that two color morphs of female side‐blotched lizards are engaged in an offspring quantity‐quality game that promotes a density‐and frequency‐dependent cycle. Orange‐throated females are r‐strategists. They lay large clutches of small progeny, which have poor survival at high density, but good survival at low density. In contrast, yellow‐throated females are K‐strategists. They lay small clutches of large progeny, which have good survival at high density. We tested three predictions of the female game: (1) orange progeny should have a fitness advantage at low density; (2) correlational selection acts to couple color alleles and progeny size; and (3) this correlational selection arises from frequency‐dependent selection in which large hatchling size confers an advantage, but only when yellow alleles are rare. We also confirmed the heritability of color, and therefore its genetic basis, by producing progeny from controlled matings. A parsimonious cause of the high heritability is that three alleles (o, b, y) segregate as one genetic factor. We review the physiology of color formation to explain the possible genetic architecture of the throat color trait. Heritability of color was nearly additive in our breeding study, allowing us to compute a genotypic value for each individual and thus predict the frequency of progeny alleles released on 116 plots. Rather than study the fitness of individual progeny, we studied how the fitness of their color alleles varied with allele frequency on plots. We confirmed prediction 1: When orange alleles are present in female progeny, they have higher fitness at low density when compared to other alleles. Even though the difference in egg size of the female morphs was small (0.02 g), it led to knife‐edged survival effects for their progeny depending on local social context. Selection on hatchling survival was not only dependent on color alleles, but on a fitness interaction between color alleles and hatchling size, which confirmed prediction 2. Sire effects, which are not confounded by maternal phenotype, allowed us to resolve the frequency dependence of correlational selection on egg size and color alleles and thereby confirmed prediction 3. Selection favored large size when yellow sire alleles were rare, but small size when they were common. Correlational selection promotes the formation of a self‐reinforcing genetic correlation between the morphs and life‐history variation, which causes selection in the next density and frequency cycle to be exacerbated. We discuss general conditions for the evolution of self‐reinforcing genetic correlations that arise from social selection associated with frequency‐dependent sexual and natural selection.     

Selecting representative model micro-organisms

Background

The sexually transmitted disease, gonorrhea, is a serious health problem in developed as well as in developing countries, for which treatment continues to be a challenge. The recent completion of the genome sequence of the causative agent, Neisseria gonorrhoeae, opens up an entirely new set of approaches for studying this organism and the diseases it causes. Here, we describe the initial phases of the construction of an expression-capable clone set representing the protein-coding ORFs of the gonococcal genome using a recombination-based cloning system.

Results

The clone set thus far includes 1672 of the 2250 predicted ORFs of the N. gonorrhoeae genome, of which 1393 (83%) are sequence-validated. Included in this set are 48 of the 61 ORFs of the gonococcal genetic island of strain MS11, not present in the sequenced genome of strain FA1090. L-arabinose-inducible glutathione-S-transferase (GST)-fusions were constructed from random clones and each was shown to express a fusion protein of the predicted size following induction, demonstrating the use of the recombination cloning system. PCR amplicons of each ORF used in the cloning reactions were spotted onto glass slides to produce DNA microarrays representing 2035 genes of the gonococcal genome. Pilot experiments indicate that these arrays are suitable for the analysis of global gene expression in gonococci.

Conclusion

This archived set of Gateway^® entry clones will facilitate high-throughput genomic and proteomic studies of gonococcal genes using a variety of expression and analysis systems. In addition, the DNA arrays produced will allow us to generate gene expression profiles of gonococci grown in a wide variety of conditions. Together, the resources produced in this work will facilitate experiments to dissect the molecular mechanisms of gonococcal pathogenesis on a global scale, and ultimately lead to the determination of the functions of unknown genes in the genome.