The endosymbiont (Buchnera sp.) of the aphid Diuraphis noxia contains plasmids consisting of trpEG and tandem repeats of trpEG pseudogenes.

Most aphids are dependent for their survival on prokaryotic endosymbionts assigned to the genus Buchnera. Among the functions of Buchnera species is the synthesis of tryptophan, which is required by the aphid host. In Buchnera species from the aphid Diuraphis noxia, the genes for anthranilate synthase (trpEG) were found on a plasmid which consisted of seven tandem repeats of a 3.2-kb unit and one 2.6-kb unit which differed in containing a 0.6-kb deletion. One of the 3.2-kb units contained open reading frames corresponding to trpEG; the remaining units contained trpEG pseudogenes (psi). The nucleotide sequence upstream of trpE contained a region that has characteristics of an origin of replication (ori). Relative to trpB (a chromosomal gene), there were about two copies of the trpEG-containing plasmid. Comparisons of the nucleotide sequences of the 3.2-kb units containing trpEG and psi trpEG indicated that most changes occurred in a 700-nucleotide segment that included the region upstream of trpE and the portion of this gene coding for the N terminus. The consequence of these changes was the silencing of trpEG by inactivation of the putative promoter region and premature termination of the TrpE peptide. In contrast, the nucleotide sequence of the segment corresponding to ori was conserved in the units containing trpEG and psi trpEG. We offer a number of speculations on the evolutionary pressure in this lineage which resulted in the silencing of most of trpEG while still retaining the regions resembling ori.     

The tryptophan biosynthetic pathway of aphid endosymbionts (Buchnera): Genetics and evolution of plasmid-associated anthranilate synthase (trpEG) within the aphididae

The bacterial endosymbionts (Buchnera) from the aphids Rhopalosiphum padi, R. maidis, Schizaphis graminum, and Acyrthosiphon pisum contain the genes for anthranilate synthase (trpEG) on plasmids made up of one or more 3.6-kb units. Anthranilate synthase is the first as well as the rate-limiting enzyme in the tryptophan biosynthetic pathway. The amplification of trpEG on plasmids may result in an increase of enzyme protein and overproduction of this essential amino acid, which is required by the aphid host. The nucleotide sequence of trpEG from endosymbionts of different species of aphids is highly conserved, as is an approximately 500-bp upstream DNA segment which has the characteristics of an origin of replication. Phylogenetic analyses were performed using trpE and trpG from the endosymbionts of these four aphids as well as from the endosymbiont of Schlechtendalia chinensis, in which trpEG occurs on the chromosome. The resulting phylogeny was congruent with trees derived from sequences of two chromosome-located bacterial genes (part of trpB and 16S ribosomal DNA). In turn, trees obtained from plasmid-borne and bacterial chromosome-borne sequences were congruent with the tree resulting from phylogenetic analysis of three aphid mitochondrial regions (portions of the small and large ribosomal DNA subunits, as well as cytochrome oxidase II). Congruence of trees based on genes from host mitochondria and from bacteria adds to previous support for exclusively vertical transmission of the endosymbionts within aphid lineages. Congruence with trees based on plasmid-borne genes supports the origin of the plasmid-borne trpEG from the chromosomal genes of the same lineage and the absence of subsequent plasmid exchange among endosymbionts of different species of aphids. Received: 22 August 1995 / Accepted: 6 September 1995     

Cloning and sequence comparison ofAvaI andBsoBI restriction-modification systems

AvaI andBsoBI restriction endonucleases are isoschizomers which recognize the symmetric sequence 5CYCGRG3 and cleave between the first C and second Y to generate a four-base 5 extension. TheAvaI restriction endonuclease gene (avaIR) and methylase gene (avaIM) were cloned intoEscherichia coli by the methylase selection method. TheBsoBI restriction endonuclease gene (bsoBIR) and part of theBsoBI methylase gene (bsoBIM) were cloned by the endo-blue method (SOS induction assay), and the remainder ofbsoBIM was cloned by inverse PCR. The nucleotide sequences of the two restriction-modification (RM) systems were determined. Comparisons of the predicted amino acid sequences indicated thatAvaI andBsoBI endonucleases share 55% identity, whereas the two methylases share 41% identity. Although the two systems show similarity in protein sequence, their gene organization differs. TheavaIM gene precedesavaIR in theAvaI RM system, while thebsoBIR gene is located upstream ofbsoBIM in theBsoBI RM system. BothAvaI andBsoBI methylases contain motifs conserved among the N⁴ cytosine methylases.     

Novel biopolymers for drug discovery

The solid phase synthesis and generation of libraries of “unnatural biopolymers” is described. These polymers are characterized by novel backbones and building blocks, the properties of which may modify their pharmacological and folding properties. © 1994 John Wiley & Sons, Inc.     

A trophic cascade in a diverse arthropod community caused by a generalist arthropod predator

We tested the hypothesis that a generalist arthropod predator, Tenodera sinensis Saussure, could trigger a trophic cascade in an old-field ecosystem. These mantids had relatively weak effects on abundance and biomass of other carnivorous arthropods as a group. The effect of mantids on herbivores was stronger than on carnivores, mainly concentrated in Homoptera and Diptera. Herbivore load was reduced by mantids with the consequence that overall plant biomass (mainly grasses) was increased. Percapita interaction strengths between mantids and other arthropod taxa were, for the most part, weakly negative. Our study demonstrates that a significant trophic cascade can be triggered by a generalist predator even within the framework of a diverse community with relatively diffuse interactions. Received: 13 January 1997 / Accepted: 28 July 1997     

Identification and mode of action of quantitative trait loci affecting seedling height and leaf area in Eucalyptus nitens

 Regions of the genome influencing height and leaf area in seedlings of a three-generation outbred pedigree of Eucalyptus nitens have been identified. Three QTLs affecting height and two QTLs affecting leaf area were located using single-factor analysis of variance. The three QTLs affecting height each explained between 10.3 and 14.7% of the phenotypic variance, while the two QTLs for leaf area each explained between 9.8 and 11.6% of the phenotypic variation. Analysis of fully informative marker loci linked to the QTLs enabled the mode of action of the QTLs to be investigated. For three loci the QTL effect segregated from only one parent, while for two loci the QTL showed multiple alleles and the effect segregated from both parents in the pedigree. The two QTLs affecting leaf area were located in the same regions as two of the QTLs affecting height. Analysis of these regions with fully informative markers showed that both QTLs were linked to the same markers, but one had a similar size of effects and a similar mode of action for both height and leaf area, whilst the other showed a different mode of action for the two traits. These regions may contain two closely linked genes or may involve a single gene with a pleiotrophic effect on both height and leaf area. The QTL with the greatest effect showed multiple alleles and an intra-locus interaction that reduced the size of the effect. Assessment for two of the QTLs in a second related family did not show an effect associated with the marker loci; however, this was consistent with the mode of action of these QTLs and the pattern of inheritance in the second family. Received: 1 August 1996 / Accepted: 25 October 1996     

Reproduction of Diaeretiella rapae on Russian wheat aphid hosts at different temperatures

Diaeretiella rapae (M'Intosh), a polyphagous and cosmopolitan parasite of aphids, was imported from China to the USA for biological control of Diuraphis noxia (Mordwilko), a pest of small grains. We studied several aspects of its biology on D. noxia hosts in the laboratory at 3 constant temperatures, 10.0, 21.1, and 26.7 °C. Females lived significantly longer than males at all 3 temperatures, and longevity was greatest at 10 °C, followed by 21.1 and 26.7 °C. Fecundity (= number of mummies) did not differ significantly among the 3 temperatures studied. The ovarian egg-load was 129.1 ± 9.3, and was significantly affected by the size of adult females. A pre-oviposition period ± 0.26 days) was detected at 10 °C, but not at the other temperatures. Pre-imaginal survivorship was similar among temperatures, while R _o and T _c decreased with temperature, and r _m increased with temperature. The offspring sex ratio (proportion females) was lowest at 26.7 °C, and similar between 10.0 and 21.1 °C. In addition, the offspring sex ratio significantly declined with the age of the female parent. The suitability of D. rapae for colonization against D. noxia in North America is discussed in relation to its responses to temperature and the climate of its home range.