Interactions of Vesicular Stomatitis Virus Structural Proteins with HeLa Plasma Membranes

THE processes whereby nucleoprotein core particles of certain animal viruses become enveloped by and bud off from host cell membranes can be studied by preparing membrane^1,2 or “sedimentable”³ fractions from infected cells and examining them for the presence of virus proteins. We find that similar experiments designed to monitor assembly of vesicular stoma-titus virus (VSV) at sites along the plasma membranes of HeLa cells are best interpreted after first investigating the possibility that virus proteins adsorb to plasma membranes during cell fractionation and membrane isolation. In this report, we show that at 0° C the membrane protein of VSV, among other virus proteins, adsorbs to plasma membranes isolated from uninfected HeLa cells. With appropriate pulse-chase experiments, however, we are able to demonstrate the progressive association, in vivo, of VSV core protein with plasma membranes of infected HeLa cells.     

Sixteen new simple sequence repeat markers from Brassica juncea expressed sequences and their cross‐species amplification

The availability of expressed sequence data derived from gene discovery programs enables mining for simple sequence repeats (SSR), providing useful genetic markers for crop improvement. These markers are inexpensive, require minimal labour to produce and can frequently be associated with functionally annotated genes. This study presents the development and characterization of 16 expressed sequence tags (EST)‐SSR markers from Brassica juncea and their cross‐amplification across Brassica species. Sixteen primer pairs were assessed for polymorphism in all genomes of the diploid and amphidiploid Brassica species. The markers show reliable amplification, considerable polymorphism and high transferability across species, demonstrating the utility of EST‐SSRs for genetic analysis of brassicas.     

Coat colour variation in the banded palm civet Hemigalus derbyanus and in Owston's civet Chrotogale owstoni

1. Coat colour variation was studied in two viverrid species belonging to the subfamily Hemigalinae, on the basis of museum specimens and observations of live animals. 2. In both species, polymorphism and ontogenetic colour change occur, and in the Owston's civet, sexual dimorphism in coat colour was identified.     

Aphid and host‐plant genotype × genotype interactions under elevated CO2

1. Elevated CO₂ can alter plant physiology and morphology, and these changes are expected to impact diet quality for insect herbivores. While the plastic responses of insect herbivores have been well studied, less is known about the propensity of insects to adapt to such changes. Genetic variation in insect responses to elevated CO₂ and genetic interactions between insects and their host plants may exist and provide the necessary raw material for adaptation. 2. We used clonal lines of Rhopalosiphum padi (L.) aphids to examine genotype‐specific responses to elevated CO₂. We used the host plant Schedonorus arundinaceus (tall fescue; Schreb), which is capable of asexual reproduction, to investigate host plant genotype‐specific effects and possible host plant‐by‐insect genotype interactions. The abundance and density of three R. padi genotypes on three tall fescue genotypes under three concentrations of CO₂ (ambient, 700, and 1000 ppm) in a controlled greenhouse environment were examined. 3. Aphid abundance decreased in the 700 ppm CO₂ concentration, but increased in the 1000 ppm concentration relative to ambient. The effect of CO₂ on aphid density was dependent on host plant genotype; the density of aphids in high CO₂ decreased for two plant genotypes but was unchanged in one. No interaction between aphid genotype and elevated CO₂ was found, nor did we find significant genotype‐by‐genotype interactions. 4. This study suggests that the density of R. padi aphids feeding on tall fescue may decrease under elevated CO₂ for some plant genotypes. The likely impact of genotype‐specific responses on future changes in the genetic structure of plant and insect populations is discussed.     

Patterns of cryptic hybridization revealed using an integrative approach: a case study on genets (Carnivora, Viverridae, Genetta spp.) from the southern African subregion

Recent years have seen the development of molecular‐based methodologies to investigate hybridization and its impact on the evolutionary process. However, morphological characterization of hybrid zones has only scantily been considered, especially in zootaxa. Thus, the level of congruence between molecular and morphological characters when attempting to detect hybrids remains a poorly tackled area. The genets (genus Genetta) provide an ideal case study for further investigation of the respective contribution of morphology and DNA in hybrid zone characterization because (1) their morphology has recently been exhaustively explored and (2) the existence of hybrid zones in southern Africa was proposed in the literature. We assessed levels of hybridization among the southern African genets, and questioned the role of ecological factors on the hybridization patterns detected. We used an integrative approach involving nine discrete morphological characters and a diagnostic discriminant function, geometric morphometrics and sequences of cytochrome b including collection specimens. The combination of independent materials allowed us to accurately reassess the level of hybridization in southern African genets, and revealed cryptic, interspecific gene flows. Morphology unambiguously detected a low number of G. maculata × G. tigrina hybrids and rejected the hypothesis of a large intergradation zone in KwaZulu‐Natal, thus supporting the species status of the two genets. Cytochrome b analyses revealed: (1) cryptic, massive hybridization between G. tigrina and the sympatric G. felina, and (2) a trace of reticulation (one sequence) between G. tigrina and the allopatric G. genetta. The type specimen of G. mossambica Matschie, 1902 is considered to be a morphological hybrid between G. maculata and G. angolensis. Remarkably, the morphological approaches (discrete characters and morphometrics) proved complementary to conclusions derived from cytochrome b sequences. Whilst morphometrics was generally unable to accurately identify all putative hybrids, this approach revealed diagnostic cranial shape differences between recognized species as well as the cryptic G. ‘letabae’ (included in the super‐species G. maculata). Morphometrics also confirmed the diagnostic value and age dependency of discrete characters. Our integrative approach appeared necessary to the detection of cryptic hybridizations and to the comprehensive characterization of hybrid zones. The recurrent detection of hybrids exhibiting tigrina‐like coat patterns may suggest (1) asymmetric hybridization of G. tigrina males to females of other species and (2) positive selection for tigrina‐like phenotype in South African habitats, but these hypotheses will have to be further tested using other sources of evidence. Despite the precise mosaic of hybrid zones identified in southern African genets, the environmental factors that shape patterns of distribution of hybrids remain unclear. Nevertheless, in the light of our range reassessment, it appears that seasonality of precipitation and periods of annual frost may play stringent roles in the distribution of genets. The complementarity of our results based on morphology and molecules is regarded as encouraging for the further development of integrative approaches in order to better understand the complex phenomena that underlie hybridization processes. © 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 86 , 11–33.     

Towards an understanding of how phloem amino acid composition shapes elevated CO2‐induced changes in aphid population dynamics

1. The performance of foliage feeders tends to decrease under elevated CO₂, but the responses of phloem‐feeding insects have been much more equivocal. As phloem tissues are less accessible than whole‐plant tissues, much less is known about how phloem composition is altered under elevated CO₂ and the mechanisms driving changes in aphid performance. 2. In this study, the plant mechanisms underlying the performance of Rhopalosiphum padi aphids on Hordeum vulgare (barley) grown under ambient (390 ppm) and elevated (700 ppm) CO₂ were examined. We used aphid stylectomy to sample pure phloem from plants in CO₂‐controlled conditions and high‐performance liquid chromatography to analyse phloem samples for amino acid concentrations. 3. Aphid abundance significantly increased by 127% under elevated CO₂. Consequently, plant biomass decreased under elevated CO₂ in trials with herbivores present, possibly due to the increased herbivore load, but increased when aphids were absent. The intrinsic rate of population increase (r_m) was significantly higher under elevated CO₂; however, there were no statistically significant effects on aphid fecundity or development time. The concentration of individual amino acids tended to increase, although these increases were statistically significant in only a few cases. A principal components analysis revealed that the relative abundance (mol %) of those amino acids considered essential for aphids tended to increase under elevated CO₂. 4. These results indicate that CO₂ may affect nutrient translocation in plants in ways that are contrary to predictions about nitrogen metabolite responses to CO₂. Such plant biochemical responses may underlie observations of improved phloem feeder performance under elevated CO₂.     

Phylogenetic relationships within the turacos (Musophagidae)

Cytochrome b sequences were used to investigate both the systematic position of the Musophagidae and the species relationships within the family. Phylogenetic analyses (neighbour-joining, maximum likelihood and maximum parsimony) supported the hypothesis that the Musophagidae are more closely related to the cuckoos, parrots and the Hoatzin than the gamebirds. Within the family, three major clades are evident that correspond to the three subfamilies generally recognized. The 'grey turacos' and the turacin-bearing turacos form two separate monophyletic groups. Corythaixoides leucogaster appears to be basal within the subfamily Criniferinae (the 'grey turacos'). This analysis yields a hypothesis for the controversy surrounding the position of Gallirex porphyreolophus and Tauraco johnstoni, which appear to be closely related and basal to the other turacin-bearing turacos. The Musophaga species are placed within the Tauraco clade and appear to be specialized forms of typical turacos. The superspecies persa forms a monophyletic group, within which T. hartlaubi falls. This is an association that has not been suggested previously.     

Insensitivity to Gibberellin in Dwarf Wheats

The responses to GA were observed in three genetically distinctdwarf wheats and compared to that of tall controls. Height,internode number and length, dry weight, and flowering timewere greatly affected by GA in the tall varieties, and hardlyat all in the dwarfs. However, the dwarfs did respond by producingmore tillers, unlike the tall varieties which produced lesslateral growth with GA treatment. The aleurone layers of the seeds of the tall varieties and Norin10-Brevor 14 produced -amlyase in response to GA, whereas inthe other dwarfs this response was greatly diminished. The possible modes of action of the hormone in wheat is discussedin the light of these results.