Sequence of the iaa and ipt region of different Agrobacterium tumefaciens biotype III octopine strains: reconstruction of octopine Ti plasmid evolution

The TA regions of biotype III octopine/cucumopine (OC) Ti plasmids are closely related to the TL region of the biotype I octopine Ti plasmids pTiAch5 and pTi15955. Sequence analysis shows that the limited and wide host range biotype III OC TA regions are derived from a common ancestor structure which lacked the 6a gene found in the biotype I octopine TL region. The TA region of the wide host range OC Ti plasmids has conserved most of the original TL-like structure. In most wide host range OC isolates the TA-iaaH gene is inactivated by the insertion of an IS866 element. However, the TA region of the wide host range isolate Hm1 carries an intact TA-iaaH gene. This gene encodes a biologically active product, as shown by root induction tests and indole-3-acetic acid measurements.The limited host range OC Ti plasmids pTiAB3 and pTiAg57 have shorter TA regions which are derived from a wide host range TA region. The AB3 type arose by an IS868-mediated, internal TA region deletion which removed the iaa genes and part of the ipt gene and left a copy of IS868 at the position of the deleted fragment. The pTiAB3 iaa/ipt deletion was followed by insertion of a second IS element, IS869, immediately 3 of the ipt gene. pTiAg57 underwent the same iaa-ipt deletion as pTiAB3, but lacks the IS868 and IS869 elements.Analysis of the various TA region structures provides a detailed insight into the evolution of the biotype III OC strains.     

The geographic distribution and host range of Capillaria hepatica (Bancroft) (Nematoda) in Australia.

The geographic distribution, host range and prevalence of Capillaria hepatica were recorded in 4629 house mice, Mus domesticus, 263 black rats, Rattus rattus, and 58 Norway rats, R. norvegicus. The parasite was found at five localities, all in or near large towns along the coast. The two Rattus species appeared to be the primary hosts of C. hepatica in Australia. Published and unpublished data on helminth infections of Australian native mammals from 1162 murids (26 species), 3018 marsupials (67 species) and 99 monotremes (two species) were compiled. Only seven animals from three murid species were infected with C. hepatica; all were from the same rainforest in northern Queensland. C. hepatica was distributed widely, occurring in the house mouse, black rat and Norway rat on a 10,850 ha farm but there was no infection in cattle, sheep or goats (abattoir records). Also, 52 rabbits, four cats and one fox (shot samples) and 27 marsupial mice, Sminthopsis crassicaudata (museum specimens), had no sign of C. hepatica infection. Overall, the results indicate that transmission of C. hepatica to native, domestic and feral mammals is rare, presumably because of ecological constraints on egg embryonation and survival. In the light of these findings, the potential use of C. hepatica as a biological agent to control mouse plagues in Australia is discussed.     

The Streptomyces ghanaensis low copy plasmid pSG2 and its use for vector construction

A plasmid, pSG2, was isolated from Streptomyces ghanaensis and characterized by electron microscopy, buoyant density measurement, and restriction enzyme analysis. The length of 13.8 kb, single restriction sites for HindIII, EcoRV and PvuII and the possibility of deleting non-essential regions of the plasmid made pSG2 a suitable basic replicon for vector development. pSG2 has a copy number of about four. Plasmid pSG2 was fused to a pACYC184 derivative modified to harbour a thiostrepton resistance gene. The resulting plasmid, designated pSW1, is a 16.6 kb shuttle plasmid which replicates in Escherichia coli and in several Streptomyces strains, including S. ghanaensis, S. lividans and S. viridochromogenes. Replacement of a Bg/II-fragment of plasmid pSG2 by a fragment encoding thiostrepton resistance resulted in a low copy 12.2 kb Streptomyces plasmid. This plasmid, designated pSW2, is a Streptomyces broad host range plasmid.     

Utilization of fruiting trees by monkeys as analyzed from feeding traces under fruiting trees in the tropical rain forest of Cameroon

Individual trees of the food species of monkeys were identified by placing plastic tapes with an identification number on them in the tropical rain forest of Cameroon, West Africa. In order to determine the use of the feeding trees by monkeys, the ground under each of the trees was checked at least once a week to see if there were any fallen fruits or traces of feeding on fruits. Some fruit species were not fed on by either monkeys or large arboreal squirrels. Among the food species common to both the monkeys and large squirrels, a larger proportion in terms of quantity in each species was mainly eaten by the monkeys except in the case of super-abundantly fruit producing species. The monkeys and large arboreal squirrels were well segregated in their diets. Larger proportions (more than 85% for most of the monkeys' major foods) of fruits of larger sizes were made to fall on the ground by the monkeys and squirrels. The monkeys displayed a tendency to visit fruiting trees rather evenly (even rate of visit = even frequency of visit/duration of fruiting) not ignoring any area of the home range, although a small difference in this tendency was observed between the two study periods, one an abundant season and the other a poor fruiting season. On average, one associated polyspecific group of monkeys encountered only 14 fruiting trees per day. On the other hand, fruits were available all around the year, as the fruiting periods of different tree species were widely distributed around the year, or the fruiting periods of some species were very long. Although the monkeys are able to depend heavily on fruits, the quantity of fruits is not so great. The population size of monkeys is well balanced with the available food supply in the tropical rain forest of West Africa.     

Insect emergence from a shallow southern West German lake,with special reference to the parasitic host-associated water mite larvae

Insect emergence was studied with floating box traps in the littoral of Lake Mindelsee from July 1st until September 30th, 1984. The material was collected every 48 h. Out of the total catch of 6897 specimens per m², 75.1 % belonged to the Chironomidae, 4,9% to the Chaoboridae, 15,4% to the Ephemeroptera and 3,2% to the Trichoptera. Endochironomus sp., Lauterborniella agrayloides, Polypedilum pullum (Chironomidae) and Oocetis testacea (Trichoptera) are reported for the first time from Lake Mindelsee. The seasonal course of the total emergence showed two distinct peaks: one at the end of July, the other at the beginning of September. Diurnal emergence was studied on two occasions, in July and in August. Most of the insects emerged during dusk and dawn. Species composition and degree of water mite infection of the collected nematocerans were studied. Host range and specific attachment sites of the different water mite taxa could only be established on the genus or family-level, with the exception of the most common water mite species Hydrodroma despiciens. In addition to its main host Chaoborus flavicans, this mite species infested 16 chironomid species with Corynoneura sp., Cryptocladopelma viridula, Pseudochironomus prasinatus and Tanytarsus lactescens as new records. The larvae of Hydrodroma despiciens were attached almost exclusively to the thorax of the imagines. Chaoborus flavicans, and within the Chironomidae Procladius sp. of the subfamily Tanypodinae, showed the highest average mite load. Species of the subfamily Chironominae were also heavily infested (e.g. Microtendipes sp., infestation rate 76 % with a maximum mite load of 18). Within the representatives of the subfamily Orthocladiinae however, only low infestation rates occurred with maximum one larva per host. Finally, problems like the synchronization of the life-cycles of hosts and parasites and strategies of coexistence between the different water mite taxa parasitizing one host are discussed.     

The influence of habitat and behavioural interactions on the local distribution of the wrasse,Pseudolabrus celidotus

Synopsis The temperate wrasse, Pseudolabrus celidotus (Labridae), was studied in a shallow rocky reef habitat to asses how ecological and behavioural factors influence its within habitat patterns of distribution. Emphasis was placed on examining the mechanisms underlying size/age related changes in dispersion and habitat use. Juveniles (< 100 mm SL) were found to be aggregated whereas adults tended to be more randomly dispersed. This pattern appeared to be a product of both (a) ecological, and (b) behavioural factors. (a) Juveniles were closely associated with shelter (macro-algae) which was located in patches throughout the habitat. Their local distribution was probably not related to the distribution of their major prey items. Shelter dependence appears to decrease with growth and fish switched from feeding on micro-crustaceans present in macro-algae to those associated with bare rock — coralline turf areas. Adults foraged throughout larger home ranges, but fed preferentially in microhabitats where their prey items (e.g. bivalves) were most abundant. (b) Juveniles tended to aggregate into loose foraging schools. This tendency declined with age and was associated with a corresponding increase in the rate of aggressive interactions among individuals of a cohort. Consequently, individuals of the same size became more spaced out as they grew. Between cohort association or aggression was rare and apparently unimportant in determining local patterns of distribution. Many interspecific foraging associations and aggressive interactions were recorded. However, with the probable exception of interspecific territoriality by the blenny Forsterygion varium, these behaviours probably had only a transitory influence on the distribution of the wrasse.     

Geography determines genetic relationships between species of mountain pine (Pinus mugo complex) in western Europe

Aim Our aims were to test whether morphological species of mountain pines were genetically supported in the western part of the distribution range of the Pinus mugo species complex (Pinus mugo Turra sensu lato), to resolve genetically homogeneous clusters of populations, to determine historical demographic processes, and to assess the potential hybridization of mountain pines with Scots pine, Pinus sylvestris L. Location Populations were sampled in the Iberian System, the Pyrenees, the French Mont Ventoux, Vosges and Jura mountains, the German Black Forest and throughout the Alps. This corresponded to a range‐wide sampling for mountain pine sensu stricto (Pinus uncinata Ram.) and to a sampling of the western parts of the ranges of dwarf mountain pine (Pinus mugo Turra sensu stricto) and bog pine/peatbog pine [Pinus rotundata Link/Pinus × pseudopumilio (Willk.) Beck]. Methods In total, 786 individuals of P. mugo sensu lato from 29 natural populations, and 85 individuals of P. sylvestris from four natural populations were genotyped at three chloroplast microsatellites (cpSSRs). Populations were characterized for standard genetic diversity statistics and signs of demographic expansion. Genetic structure was explored using analysis of molecular variance, differentiation statistics and Bayesian analysis of population structure (BAPS). Results One hundred haplotypes were identified in P. mugo sensu lato. There was a stronger differentiation between geographical regions than between morphologically identified taxa (P. mugo sensu stricto, P. uncinata and P. rotundata/P. ×pseudopumilio). Overall genetic differentiation was weak (G_ST = 0.070) and displayed a clear phylogeographic structure [N_ST = 0.263, N_ST > N_ST (permuted), P < 0.001]. BAPS identified a Pyrenean and an Alpine gene pool, along with several smaller genetic clusters corresponding to peripheral populations. Main conclusions The core regions of the Pyrenees and Alps were probably recolonized, respectively by P. uncinata and P. uncinata/P. mugo sensu stricto, from multiple glacial refugia that were well connected by pollen flow within the mountain chains. Pinus rotundata/P. × pseudopumilio populations from the Black Forest, Vosges and Jura mountains were probably recolonized from various glacial populations that kept their genetic distinctiveness despite late glacial and early Holocene expansion. Marginal P. uncinata populations from the Iberian System are compatible with elevational shifts and long‐term isolation. The causes of haplotype sharing between P. mugo sensu lato and P. sylvestris require further research.     

World phylogeography and male‐mediated gene flow in the sandbar shark,Carcharhinus plumbeus

The sandbar shark, Carcharhinus plumbeus, is a large, cosmopolitan, coastal species. Females are thought to show philopatry to nursery grounds while males potentially migrate long distances, creating an opportunity for male‐mediated gene flow that may lead to discordance in patterns revealed by mitochondrial DNA (mtDNA) and nuclear markers. While this dynamic has been investigated in elasmobranchs over small spatial scales, it has not been examined at a global level. We examined patterns of historical phylogeography and contemporary gene flow by genotyping 329 individuals from nine locations throughout the species’ range at eight nuclear microsatellite markers and sequencing the complete mtDNA control region. Pairwise comparisons often resulted in fixation indices and divergence estimates of greater magnitude using mtDNA sequence data than microsatellite data. In addition, multiple methods of estimation suggested fewer populations based on microsatellite loci than on mtDNA sequence data. Coalescent analyses suggest divergence and restricted migration among Hawaii, Taiwan, eastern and western Australia using mtDNA sequence data and no divergence and high migration rates, between Taiwan and both Australian sites using microsatellite data. Evidence of secondary contact was detected between several localities and appears to be discreet in time rather than continuous. Collectively, these data suggest complex spatial/temporal relationships between shark populations that may feature pulses of female dispersal and more continuous male‐mediated gene flow.     

Strong genetic differentiation and postglacial origin of populations in the marine midge Clunio marinus (Chironomidae,Diptera)

The marine midge Clunio marinus (Chironomidae, Diptera) is characterized by a one‐dimensional distribution along the European Atlantic coast, where its lunar and circadian emergence rhythms are genetically adapted to the local tidal regimes, resulting in a series of ‘temporal races’. Clunio marinus is restricted to rocky coasts and thus the temporal races occur in different rocky patches. We studied 10 populations of Clunio marinus from five different regions, spanning the major rocky mainland coasts from Spain to Norway, using amplified fragment length polymorphisms (AFLP), microsatellites and mitochondrial cytochrome oxidase I (COI) sequences. Star‐like patterns of COI haplotypes within regions indicate postglacial colonization. A high degree of shared polymorphisms in AFLP markers suggests colonization from a single source, implying postglacial evolution of timing adaptations in relation to the local tidal regime. In contrast, no COI haplotypes are shared among regions. We hypothesize that different levels of differentiation of nuclear vs. mitochondrial markers in the source region were carried forward during postglacial expansion. Despite the recent origin of populations, all markers reveal distinct genetic differentiation between rocky coasts on a scale of 650–1150 km. Differentiation between rocky coasts is not correlated to timing adaptations, suggesting that geographic isolation is prevalent between rocky coasts and that this facilitated the evolution of local timing adaptations. At the same time there is little genetic differentiation within rocky coasts on a scale of 2–6 km; leaving open the possibility that within rocky coasts with large variation in tidal regimes, temporal adaptations evolved in the face of gene flow.     

Site-specific recombination of T2 phage using IP008 long tail fiber genes provides a targeted method for expanding host range while retaining lytic activity

The application of bacteriophages (phages) in therapy urgently requires the production of wide-host-range recombinant phages that possess strong lytic activity. The wide-host-range IP008 phage was classified by transmission electron microscopy analysis as an A2 morphotype member of the Myoviridae family of the order Caudovirales . IP008 showed a high homology (99.4% similarity in the amino acid alignment of the major capsid protein Gp 23) with KEP10, another wide-host-range phage. The long tail fiber genes (genes 37 and 38 ) from the genome of T2 were replaced with those of the IP008 phage by homologous recombination. The host range of the recombinant phages was identical to that of IP008. Furthermore, the recombinant phage bacterial lytic activity was restored. Future analyses of host-range mutants of the closely related phages T2 and IP008 could lead to a more precise localization of the genetic factors responsible for receptor specificity.