Biological and biochemical properties of an isolate of cherry rasp leaf virus from red raspberry

A mechanically transmissible virus obtained from symptomless plants of a red raspberry selection imported into Scotland from Quebec, Canada was indistinguishable serologically from a cherry isolate of cherry rasp leaf virus (CRLV). The raspberry isolate, CRLV-R, was graft transmitted to several virus indicator species and cultivars of Rubus without inducing noticeable symptoms. In Chenopodium quinoa sap, CRLV-R lost infectivity after dilution to 10^-5 or heating for 10 min at 60°C but was infective after 16 days (the longest period tested) at 18°, 4° or - 15°C. The virus particles are isometric, c. 28 nm in diameter, and were purified with difficulty from infected C. murale and C. quinoa plants. The particles comprise two nucleoprotein components with sedimentation coefficients of 89 and 115 S and are prone to aggregate during purification. When centrifuged to equilibrium in CS₂SO₄ solution, purified virus preparations formed two major components with p= 1·28 and 1·36 g/cm³. Virus particles contained two RNA species which, when denatured in glyoxal and electrophoresed in agarose gels, had estimated mol. wt of 2·56 × 10⁶ (RNA-1) and 1·26 × 10⁶ (RNA–2). Infectivity of CRLV-R RNA was abolished by treatment with proteinase K, suggesting that the RNA is linked to protein necessary for infectivity; RNA molecules contained polyadenylate. In reticulocyte lysates, CRLV-R RNA stimulated the incorporation of ³H-leucine, mainly into two polypeptides of estimated mol. wt 200 000 and 102 000. When electrophoresed in polyacrylamide gels, protein obtained from CRLV-R particles purified by centrifugation to equilibrium in Cs₂SO₄ separated into three bands with estimated mol. wt 26 000 , 23 000 and 21 000.     

Characterization of a Neutral Aminoacyl-Peptide Hydrolase from Naegleria fowleri1

An intracellular alpha-aminoacyl-peptide hydrolase (EC 3.4.11.-) from Naegleria fowteri nN68 (ATCC 30894) has been characterized. The enzyme preparation hydrolyzed phenylalanyl-, tyrosyl-, leucyl-, arginyl-, alanyl-, tryptophanyl-, histidyl-, methionyl-, and lysyl-naphthylamide but not benzoylleucyl-, leucylglycyl-, glycylprolylleucyl-, glycyl-, threonyl-, aspartyl-, or glutamyl-naphthylamide. The aminopeptidase activity was inhibited by the cysteine-protease inhibitors—hydroxymercuribenzoate, chloromercurisulfate, and iodoacetate- by the aminopeptidase inhibitors-bestatin and trans-epoxysuccinyl-leucyl-agmatine- by an inhibitor of soluble alanyl aminopeptidase EC 3.4.11.14, puromycin, and by the metalloprotease inhibitor, o-phenanthroline. The exopeptidase activity was not inhibited by the chelator, ethylenediaminetetraacetate, or the serine-protease inhibitor, phenylmethylsulfonylfluoride. The pH optimum of the exopeptidase was between 7.0 and 8.0. Enzyme activity was stable at 55°C for 30 min, but all activity was lost after 15 min at 80°C. Enzyme activity was inhibited by 100 μM HgCI₂ and CdCl₂ but not by 1 mM CoCl₂, CuCl₂, MnCl₂, NiCl₂, FeCl₂, or ZnCl₂. Enzyme activity was inhibited by 0.1% sodium dodecyl sulfate but not by 0.2% Brij 35, Tween 20, Tween 80, or Triton X-100.     

Behavioural response of spider mites (Tetranychus urticae) to induced resistance of cotton plants

ABSTRACT.

• 1 Spider mites (Tetranychus urticae Koch) given a choice between a cotton plant previously damaged by mites and an undamaged control preferentially moved to the control plant.
• 2 This host-choice behaviour was seen in adult female mites but not in immatures.
• 3 Adult females were not found to be significantly more fecund on control plants than on previously damaged plants, but the duration of this choice experiment precluded full assessment of effects on fecundity.
• 4 Mites responded very quickly, choosing previously undamaged plants without contacting or feeding on the test plants. This suggests that an olfactory response to a chemical substance(s) is involved.

     

Synthesis of Substance P

SUBSTANCE P has been synthesized by the solid-phase procedure of Merrifield^1,2 according to the sequence H-Arg-Pro-LysPro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH₂ reported in the previous letter.     

Ultrastructure of Acrasis rosea,a Cellular Slime Mold,During Development*

SYNOPSIS. An ultrastructural study of the myxamoebae of Acrasis rosea in the vegetative, aggregative and culminative stages was made. An intracytoplasmic system of microfibrillar bundles develops as the cells enter the aggregative stage and commence the morphogenetic sequence leading to the construction of a fruiting body. The fibrillar bundles disappear in the cells of the mature fruiting body. No relevant ultrastructural differences were observed between spores, stalk cells and microcysts. Each of these cells is surrounded by a single-layered coat of fibrillar material that is oriented parallel to the cell surface. Tubular structures were observed between the plasma membrane and the cell coat. The tubules may be layered along the cell periphery or they may be recessed in pockets formed by the plasma membrane. They resemble lomasomes typical of fungal cells. The myxamoebae of A. rosea clearly differ from the Dictyostelium-type myxamoebae in mitochondrial structure, the presence of lamellate structures in the nucleolus and the absence of prespore vacuoles.     

THE USE OF A MULTIPLE-TRANSFER METHOD IN PLANT VIRUS TRANSMISSION STUDIES—SOME STATISTICAL POINTS ARISING IN THE ANALYSIS OF RESULTS

The frequency of success of attempts to transmit a virus disease from one plant to another has long been used to measure the effects of any of the factors which influence transmission. Samples are taken from the population under test (e.g. vectors, diseased plants, etc.), and usually one sample is tested on each test plant; this is binomial sampling. However, in the procedure we name the 'multiple-transfer method' more than one sample may be tested on each test plant. This increases the number of samples tested without increasing the number of test plants used, and errors due to heterogeneity in the population under test are therefore minimized. Results from experiments using the multiple-transfer method may be evaluated by using the maximum likelihood estimator. The method is particularly reliable when the proportion of infected samples being studied is small, but can lead to considerable over-estimation when the proportion is high.     

Pollen movement to flowering canopies of pistillate individuals of three rain forest tree species in tropical Australia

This study measured the quantities of effective pollen vectors and their pollen loads arriving at the canopies of dioecious tropical rain forest trees in north-east Queensland. Population flowering synchrony, effective pollinator populations and pollen loads transferred between staminate and pistillate trees were compared among three insect-pollinated tree species. All three were visited by a wide range of insects, 75% of which (mostly 3–6 mm long) carried conspecific pollen. Fewer than 8% of individual insects were found to be carrying single-species pollen exclusively and none could be described as specialist pollen foragers. The introduced honeybee carried greater quantities of pollen than any native species but was not necessarily a reliable pollinator. The brief flowering periods in Neolitsea dealbata (3–4 weeks) and Litsea leefeana (4–5 weeks) populations were synchronized among individuals. Flowering in the Diospyros pentamera population extended over 15 weeks and most individuals were in flower for most of this period. Staminate trees began flowering earlier, produced more flowers and attracted relatively more insects than did pistillate trees, suggesting a density-dependent response of pollinators to flowering performance. Pollen was trapped in greater quantities on insects at staminate trees than at pistillate trees. Insect numbers increased at peak flowering periods and Diptera were the most abundant flower visitors. Anthophilous Coleoptera were more numerous at staminate than at pistillate trees in all three tree species populations. Larger quantities of pollen were mobilized during peak flowering times although the greatest quantities were transferred to pistillate canopies towards the end of the population flowering periods. Diptera carried pollen more often to pistillate N. dealbata and L. leefeana trees than did other groups whereas Coleoptera carried pollen more often to pistillate D. pentamera trees. The two contrasting flowering performances in the three tree species are discussed with reference to mechanisms that facilitate pollen transfer between staminate and pistillate trees.     

Structure and Function in Vertebrate Skeletal Muscle

SYNOPSIS. The functional diversity of vertebrate skeletal musclelargely depends upon its structure. An important aspect of thisis its hierarchical design. At the cellular level, muscle fibersform three categories whose functional properties grade intoeach other: slow-oxidative fibers with high endurance to fatigue,fast-oxidative\glycolytic fibers also endurant but with greatermetabolic diversity, and fast-glycolytic fibers with limitedendurance but quick response. This partitioning of functionalproperties found among single muscle fibers is conserved ata second level of the structural hierarchy, since the groupof myofibers innervated by a single motor neuron (together calleda motor unit) is composed of the same fiber type. Differentmotor units may be recruited in an orderly pattern dependingupon the functional demands of a particular behavior. Finally,groups of motor units innervated by axons travelling togetherin the primary nerve branches may form discrete neuromuscularcompartments at a third level of structural hierarchy. Differentmotor units may be found in regional arrays in these compartmentsso that slow or fast units tend to be clumped together and maybe recruited together as larger functional units. This hierarchicalorganization of skeletal muscle may be a fundamental vertebrateplan that allows the diversity of functions so evident in vertebratebehavior.     

Past vegetation changes in the Brazilian Pantanal arboreal–grassy savanna ecotone by using carbon isotopes in the soil organic matter

Measurements of the organic carbon inventory, its stable isotopic composition and radiocarbon content were used to deduce vegetation history from two soil profiles in arboreal and grassy savanna ecotones in the Brazilian Pantanal. The Pantanal is a large floodplain area with grass-dominated lowlands subject to seasonal flooding, and arboreal savanna uplands which are only rarely flooded. Organic carbon inventories were lower in the grassy savanna site than in the upland arboreal savanna site, with carbon decreasing exponentially with depth from the surface in both profiles. Changes in ¹³C of soil organic matter (SOM) with depth differed markedly between the two sites. Differences in surface SOM ¹³C values reflect the change from C₃ to C₄ plants between the sites, as confirmed by measurements of ¹³C of vegetation and the soil surface along a transect between the upland closed-canopy forest and lowland grassy savanna. Changes of ¹³C in SOM with depth at both sites are larger than the 3–4 per mil increases expected from fractionation associated with organic matter decomposition. We interpret these as recording past changes in the relative abundance of C₃ and C₄ plants at these sites. Mass balances with ¹⁴C and ¹³C suggest that past vegetational changes from C₃ to C₄ plants in the grassy savanna, and in the deeper part of the arboreal savanna, occurred between 4600 and 11 400 BP, when major climatic changes were also observed in several places of the South American Continent. The change from C₄ to C₃, observed only in the upper part of the arboreal savanna, was much more recent (1400 BP), and was probably caused by a local change in the flooding regime.     

Genetic analysis of populations of threatened snake species using RAPD markers

Snakes are a particularly threatened vertebrate taxon, with distributions of many species and populations becoming increasingly fragmented. At present, little is known about the degree of genetic differentiation that exists between isolated populations even though such information may be critical to their survival and conservation. As an example of how recently developed RAPD genetic markers can be used in conservation genetics, we present preliminary results from a study which used these DNA-based markers to assess population divergence in two threatened Canadian snakes, the black rat snake ( Elaphe o. obsoleta ) and the eastern massasauga rattlesnake ( Sistrurus c. catenatus ). We present information on the levels of variation and reliability of amplification for fragments generated from five primers. We then use a recently developed analytical technique to estimate levels of nucleotide diversity within populations and sequence divergence between populations. Our results show that intrapopulation levels of divergence as estimated by the methods of Clark & Lanigan ( Molecular Biology and Evolution 1993, 10 , 1096–1111) approximate those found for mtDNA in vertebrates and that diversity between snake populations is small and non-significant when tested using randomization procedures. Thus, our study provides an example of how RAPDs can be applied to conservation genetic studies of vertebrates and suggest that the snake populations we examined have only recently become isolated and maybe considered genetically equivalent from a conservation perspective, although this conclusion needs to be confirmed with other DNA-based markers.