Forage and bed sites characteristics of Indian muntjac (<Emphasis Type="Italic">Muntiacus muntjak</Emphasis>) in Hainan Island,China

Microhabitat factors associated with forage and bed sites of Indian muntjac (Muntiacus muntjak) were examined and compared in Hainan Island from 2001 to 2002. Habitat characteristics at forage and bed sites were measured by tracking five radio-collared adult muntjacs (three females, two males) and locating from fresh sign or muntjac flushed during transect surveys. Among six available habitat types, Indian muntjac preferred shrub grassland and thorny shrubland, and used dry savanna in proportion to its availability. Muntjac avoided woods, cultivated grassplot and deciduous monsoon forests. Comparing forage sites with bed sites, food availability was greater at forage sites. For bed sites, taller trees with larger canopies, taller shrubs, denser shrub canopy covers and concealment covers were essential factors. No seasonal difference (wet season vs. dry season) was found in food abundance at either forage sites or at bed sites. During the wet season, canopy closure for both types of sites was higher and at bed sites concealment cover was higher than during the dry season. Principal component analysis indicated that tree height, d.b.h. and maximum canopy diameters were important factors in habitat selection. The different microhabitat characteristics at forage and bed sites can be a clue to understanding the survival strategy of Indian muntjac, a small-bodied ungulate, in savanna woodlands.     

Acoustic structure of alarm calls in Indian sambar (<Emphasis Type="Italic">Rusa unicolor</Emphasis>) and Indian muntjac (<Emphasis Type="Italic">Muntiacus vaginalis</Emphasis>) in South Vietnam

The alarm call acoustic structure and nonlinear vocal phenomena of the Indian sambar (Rusa unicolor) and northern Indian muntjac (Muntiacus vaginalis) have been analyzed in detail as well as their vocal behavior in response to mobbing humans under natural conditions of southern Vietnam. The alarm calls of sambars, tonal barks separated by large intervals, were produced by animals standing on the place and gazing at a potentially dangerous object. Muntjacs flee off in danger and produced a series of dull barks interrupted with short intervals from a distance. The alarm call frequencies were characterized for sambars and muntjacs. The results of our study have been compared with the published data on alarm calls of other Cervidae species.     

The effect of environmental gradients on the bed site selection of roe deer (<Emphasis Type="Italic">Capreolus capreolus</Emphasis>)

A wildlife species’ selection of bedding sites is often characterised by strong trade-offs, as habitat quality, predator avoidance and foraging needs should be achieved simultaneously. Human activities often represent major threats in addition. In areas of intensive agriculture, e.g. mowing is one of the main causes of mortality of roe deer (Capreolus capreolus) fawns due their hiding strategy. For a species’ offspring, the selection of bedding sites is particularly crucial and thus, identifying how and when animals use such habitats is important for management. We used a long-term dataset of marked roe deer fawns in Switzerland (1971–2015) to reveal the characteristics of optimal bedding sites within the first two weeks of a fawn’s life in three contrasting landscapes and the potential trade-offs that may occur. We hypothesised that roe deer adjust the selection of bedding sites to current environmental conditions and available habitat to achieve sufficient levels of predator avoidance and thermoregulation necessary for the fawn’s survival, as well as the availability of sufficient food resources for the mother doe. We found that, in general, grassland habitats with medium vegetation height (20–50 cm) and habitats in close proximity to the edge of the forest were favoured to achieve those basic requirements. However, the use of bed site habitats differed between the three contrasting landscapes in dependence of elevation and hence vegetation phenology. Our results provide essential information to reduce mortality rates caused by mowing and improve the reproductive success of this species.     

Oviposition Sites of the Cypress Seed Bug <Emphasis Type="Italic">Orsillus maculatus</Emphasis> and Response of the Egg Parasitoid <Emphasis Type="Italic">Telenomus</Emphasis> gr. <Emphasis Type="Italic">Floridanus</Emphasis>

Phytophagous insects have many strategies to escape parasitoids, for example by hiding eggs into plant tissues, but oviposition in holes made by another insect is rather scarce. The cypress seed bug Orsillus maculatus Fieber (Heteroptera: Lygaeidae) is strictly dependent on the availability of cones of Cupressus sempervirens L. to oviposit. Females lay eggs either in exit holes cut through the cone scale by emerging adults of the cypress seed chalcid, Megastigmus wachtli Seitner (Hymenoptera: Torymidae), or on the inner side of scales of partly open cones. A recently discovered egg parasitoid belonging to the genus Telenomus has been shown to attack bug eggs in both oviposition sites. In this paper we investigated the parasitoid performance according to oviposition sites. Field samplings were conducted in two evergreen cypress orchards located in the south of France. The distribution and condition of the egg patches were compared between the two locations and oviposition sites. Seed bugs preferred to oviposit in emergence holes of M. wachtli, and parasitoid performance was higher in eggs laid on cone scales. The chalcid emergence holes seemed to ensure bug eggs with enemy-free space. Oviposition site selection could be an adaptive strategy to escape parasitoid attack.     

Habitat overlap between sympatric European hares (<Emphasis Type="Italic">Lepus europaeus</Emphasis>) and Eastern cottontails (<Emphasis Type="Italic">Sylvilagus floridanus</Emphasis>) in northern Italy

In northern Italy, the range of the Eastern cottontail (Sylvilagus floridanus) largely overlaps with that of the native European hare (Lepus europaeus) on the Po Plain. Both species appear to have similar habitat requirements. We studied habitat selection by hares and cottontails during feeding activity from September 2006 to August 2007 in two areas where they occur alone (allopatry) and in one area where they occur together (sympatry). The three areas were basically similar, so that shifts in habitat use observed in sympatry should reflect the response to interspecific competition. Habitat selection was examined at micro- and macro-habitat levels throughout seasons. Habitat breadth of both species followed the change of resource availability through seasons in allopatry as well as in sympatry. No shifts in habitat use were evident at macro-habitat level, even during autumn which was the limiting season. Exploitation of shared habitats by the two species seems to be promoted by differential micro-habitat use within macro-habitat types. Cottontails used woods with dense understory in greater proportion than hares, and their present sites were concentrated within the maximum distance of 20 m of the nearest shelter site. Hares were more likely than cottontails to exploit crops, and their sites were distributed even greater than 80 m away from permanent cover patches. The habitat heterogeneity of agricultural ecosystems within the sympatry range could buffer the negative effects of external factors (climate, human disturbance and predation) on hares, and enhance the chances of exploitation of shared habitats by both species.     

The evolutionary history of PDR in <Emphasis Type="Italic">Brachypodium</Emphasis><Emphasis Type="Italic">distachyon</Emphasis> polyploids

The ATP-binding cassette transporter genes include the pleiotropic drug resistance (PDR) family found only in fungi and plants. These transporters transport toxic compounds across biological membranes. Here, we investigated the evolution of the PDR1 gene in Brachypodium distachyon, a widely distributed temperate grass species that belongs to the Poaceae (Gramineae) family, which also contains the domesticated cereal crops. Because this species has multiple ploidy levels, investigating PDR1 evolution in B. distachyon will offer insights into the formation and evolution of polyploidy. From 23 B. distachyon ecotypes, 39 PDR1 homologs were identified. All ecotypes had either one or two PDR1 copies. Based on restriction site analysis, the PDR1 homologs were classified as E or H type. All but one diploid and tetraploid ecotypes had only a single H type PDR1. All but one hexaploid ecotypes had both an E and a H type PDR1. Phylogenetic analysis revealed that each type formed a well-supported cluster. The two PDR1 types appeared to evolve differently. These different evolutionary patterns could indicate a difference in age between the two types or might indicate different mutation rates or selection pressures on the two types. The phylogenetic analysis also revealed that the hexaploid ecotypes shared a genomic origin for their E type PDR1, but there were multiple origins for hexaploid H type PDR1 homologs. Overall, the results suggest that tetraploid and hexaploid might be misnomers in B. distachyon and suggest a complex polyploidization history during B. distachyon evolution.     

<Emphasis Type="Italic">Agrobacterium</Emphasis>-Mediated Transformation of Common Bermudagrass (<Emphasis Type="Italic">Cynodon dactylon</Emphasis>)

Common bermudagrass, Cynodon dactylon, is a widely used warm-season turf and forage species in the temperate and tropical regions of the world. We have been able to transform the species using Agrobacterium-mediated approach. In seven experiments reported here, a total of 67 plates of calluses and suspensions were infected with Agrobacterium tumefaciens strains, and nine hygromycin B resistant calluses were obtained after selection. Among them two green independent transgenic plants were recovered. The plants growing in pots looked relatively compact at the beginning, but the ploidy level of the plants, as determined by nuclear DNA content, was not altered.     

The influence of wild boar <Emphasis Type="Italic">(Sus scrofa)</Emphasis> on microhabitat quality for the endangered butterfly <Emphasis Type="Italic">Pyrgus malvae</Emphasis> in the Netherlands

The decline of open habitats in Europe, such as semi-natural grasslands and heathlands, has caused a general decline in biodiversity, which has been well documented for butterflies. Current conservation practices often involve grazing by domestic livestock to maintain suitable butterfly habitats. The extent to which wild ungulates may play a similar role remains largely unknown. Through their rooting activity, wild boar could be effective to reduce grass encroachment and restore pioneer microhabitats that are vital to many grassland insects in temperate climates. Here, we assessed the microhabitat requirements of Pyrgus malvae, an endangered butterfly of heathland and grassland habitats in the Netherlands, with special attention for the influence of wild boar rooting. To date, oviposition site selection of this species has concentrated on calcareous grasslands, whereas we also include heathlands. Overall, larval occupancy was higher in warm, open and sparsely vegetated microhabitats, which supports earlier findings. In heathland, microhabitat occupancy was positively affected by bryophyte and litter cover. In heath-grassland mosaic, microhabitat occupancy was also influenced by bryophyte and litter cover, but in addition low grass cover increased occupancy by favouring host plants. In grassland, only low grass cover and host plant cover determined microhabitat quality. Across all habitats, occupied microhabitats were characterized by lower vegetation as well as higher average daytime temperatures than unoccupied microhabitats. We discovered that wild boar play an important role in reducing grass cover by shallow rooting in grass patches, thereby increasing host plant availability. Hence, wild boar may have an added value in maintaining and restoring P. malvae microhabitats in grassland habitats in addition to grazing by domestic livestock.     

Predicting the status of wild deer as hosts of <Emphasis Type="Italic">Mycobacterium bovis</Emphasis> infection in Britain

Control of livestock diseases can become complicated when wild animals are involved. The Eurasian badger (Meles meles) is considered the principle wildlife host of Mycobacterium bovis (which causes bovine tuberculosis, bTB) in Great Britain and Ireland, but wild deer have also been implicated. Whether wild deer are likely to perpetuate bTB in cattle depends on the exposure risks they pose, the mode of pathogen transmission, the distances over which the disease can be transported and whether they can maintain infection within their own populations independently of other sources. We evaluated the likely host status of each of four species of wild British deer (red, roe, fallow and Reeves' muntjac) and the badger across a range of densities typically observed in Britain by manipulating the reproductive number equation proposed by Anderson and May (1991). We estimate that roe deer almost certainly act as spillover hosts at densities lower than 30 km⁻², red deer below 16 km⁻², muntjac below 6 km⁻², fallow below 4 km⁻² and the badger below 2 km⁻². We also estimate that muntjac will almost certainly act as maintenance hosts at densities above 56 km⁻², fallow above 47 km⁻² and badgers above 24 km⁻². For densities between these values, we cannot be certain of the host status of these species, and for red and roe deer we cannot be certain of host status under most natural conditions typically experienced in parts of Britain experiencing high incidence of bTB in cattle. However, enhanced transmission rates resulting from artificially high densities such as might be experienced at supplementary feeding sites may be sufficient to promote independent maintenance of infection. We were not able to confidently assign host status to any species over a wide range of densities, but conclude that this is likely to reflect reality, where host status may be affected as much by, for example, demographic fluctuations as it is by population density. Our results imply densities below which populations of wild deer inhabiting cattle bTB hotspots ought to be maintained in order to control the possibility of them perpetuating the cycle of intra- and interspecific M. bovis transmission.     

Species differentiation and gene flow in the Blackbutts (genus <Emphasis Type="Italic">Eucalyptus</Emphasis> subgenus <Emphasis Type="Italic">Eucalyptus</Emphasis> section <Emphasis Type="Italic">Pseudophloius</Emphasis>)

The significance of the taxonomic distinction of two species of Blackbutt was studied by analysing patterns of genetic (microsatellite markers; n = 13) and phenetic (capsule morphology) differentiation. Analysis of genetic structure using a Bayesian modelling approach on range-wide samples of both taxa (n = 457) showed the major division was within the more widely distributed species, Eucalyptus pilularis, and not aligned with taxonomy. Comparisons of intra- and inter-taxon genetic differentiation in paired-samples of taxa from each of four locations spanning the distribution of the more restricted E. pyrocarpa, showed that around twice as much variation was found among locations within taxa, than between taxa. Despite the lack of differentiation at effectively neutral microsatellite markers, significant phenetic differences (including capsule size) were evident between taxa at most sites. A landscape mosaic of taxa, coincident with changes in elevation, vegetation and soil types, suggested some phenetic differences were probably adaptive and spatial differentiation was stabilised by environmental factors. An absence of morphological intermediates and a lack of correlation in the rankings of locus inter-taxon differentiation (Phi_BT) across locations, was consistent with parapatric origins for E. pyrocarpa. We conclude the taxa are at the lower end of the speciation spectrum and might best be viewed as ecotypes, divergent in evolutionary potential, but with genomes broadly permeable to inter-taxa gene flow. Gene exchange between plantings of E. pilularis and nearby E. pyrocarpa forest is likely as the two taxa appear to have few barriers to reproduction.     

The Mating System of <Emphasis Type="Italic">Amegilla</Emphasis> (<Emphasis Type="Italic">Asarapoda</Emphasis>) <Emphasis Type="Italic">paracalva</Emphasis> Brooks (Hymenoptera: Apidae)

Males of the solitary bee Amegilla (Asarapoda) paracalva employ two mate-locating tactics: aggressive defense of sites from which virgin females are emerging and patrolling flower patches that are visited by conspecific females. At one study site, a single male was able to control an entire emergence area for one or more days. Multiple males patrolled one flower patch, interacting aggressively on occasion but no one individual was able to monopolize this resource. Territorial males at the emergence site secured mates by waiting by tunnels for receptive virgin females to emerge after metamorphosis. Males patrolling the flower patch pounced upon flower visiting conspecifics and mated with receptive females there. Territorial males at the emergence site were larger than average individuals, probably because of the advantage larger males have when grappling with opponents. Flower patrolling males were smaller than territorial males at the emergence sites, perhaps because of the advantages gained by these males from rapid, agile flight.     

<Emphasis Type="Italic">Drosophila</Emphasis> pupation behavior in the wild

We investigated pupa distributions of D. simulans, D. buzzatii, D. melanogaster, D. immigrans and D. hydei on a number of natural breeding sites. Pupae of all five species showed aggregated distributions, which prompted us to examine these aggregations in a more detail for two species that commonly co-occur in breeding sites, D. simulans and D. buzzatii. We found that pupae of both species tend to be aggregated in conspecific clusters. Subsequent experiments revealed that both species are attracted to the odors of other larvae, though only D. buzzatii differentiated between conspecifics and heterospecifics (they preferred conspecific). Furthermore, third instar larvae of both species preferred more alkaline substrates. Altogether, our results demonstrate that Drosophila species form conspecific pupa aggregations in natural breeding sites, and that pupation site selection depends on interactions among conspecific and heterospecific larvae and on chemical characteristics of the breeding sites.     

Digestive responses of two omnivorous rodents (<Emphasis Type="Italic">Peromyscus maniculatus</Emphasis> and <Emphasis Type="Italic">P. alstoni</Emphasis>) feeding on epigeous fungus (<Emphasis Type="Italic">Russula occidentalis</Emphasis>)

The sporocarps of hypogeous and epigeous fungi are important dietary items for forest dwelling rodents in temperate and tropical forests throughout the world. However, results of some pioneering works have demonstrated that fungi cannot be considered as nutritionally high-quality food items for some mycophagous small rodents. According to these studies, when mycophagous rodents feed on fungus, they showed a minimal digestibility, but whether this applies to most rodent species that include fungi in their diets is unknown. In this study, we experimentally evaluated body mass changes and feed preferences in captive deer (Peromyscus maniculatus) and volcano (P. alstoni) mice when fed on epigeous fungus (Russula occidentalis). In experiment 1, the animals were fed with fungus as the only feedstuff in comparison to regular rodent chow and oat. In experiment 2, the animals were fed with fungus in a free-choice arrangement together with equal amounts of rodent chow and oat. Both species lost ∼15% of their body mass within 4 days when fed on fungus alone, but gained 5–10% body mass during the same time period when ingesting oat and rodent chow, respectively, as the only feedstuff. However, in contrast, in the free-choice arrangement with all three feedstuffs, both species gained 20–30% body mass, and showed the highest feed preference for fungus followed by oat and rodent chow. In addition, apparent digestibility of energy and nitrogen were analyzed in both rodent species, which were 50–60% for fungus, whereas approximately 90–94% for rodent chow and oat. According to our results, animals need to supplement their diets with alternative high-quality food items in order to maintain and increase their body mass, suggesting that epigeous fungi are only of moderate nutritional value for small rodents. Futures studies should focus on exploring the importance of a mixture of fungal species in the diet of small mycophagous rodents.     

Sleeping Sites of <Emphasis Type="Italic">Rhinopithecus brelichi</Emphasis> at Yangaoping,Guizhou

Arboreal primates spend about half of their lives at sleeping sites; hence, selection of sleeping sites is crucial for individual survival, and data concerning them is important for conservation efforts. We collected data on sleeping sites for a group of the endangered snub-nosed monkey (Rhinopithecus brelichi) at Yangaoping (27°58′N, 108°45′E) from January 2006 to December 2007. All sleeping sites were located in the mid-slope and in the shadow of ridges facing the northeast and southeast. The monkeys remained quiet while entering and occupying sleeping sites, and slept in evergreen species during the cold season (December–March). Trees in sleeping sites were similar in height and girth at breast height to those elsewhere, but some trees in lower areas were larger. The monkeys usually slept in close proximity to the last feeding spot, and their daily activities usually occurred around the sleeping site. Areas adjacent to sleeping sites were used more intensively than those not adjacent. Monkeys left the sleeping sites later in the morning in the cold season. These behavioral responses suggested that predation risk, thermoregulation, and climate stresses are the main determining factors in the selection of sleeping sites for this temperate monkey.     

Co-invasion of three Asian earthworms, <Emphasis Type="Italic">Metaphire hilgendorfi</Emphasis>, <Emphasis Type="Italic">Amynthas agrestis</Emphasis> and <Emphasis Type="Italic">Amynthas tokioensis</Emphasis> in the USA

Earthworm invasions are one of the most serious causes of ecological deterioration in the temperate deciduous forests of North America. Non-native earthworms impact understory vegetation, leaf litter layer, carbon dynamics, nutrient availability, and the associated food webs. Here we report a significant status change and confirm expansion of known range of Amynthas agrestis, one of the most serious invasive species in North America, and two of its close relatives, A. tokioensis and Metaphire hilgendorfi. The three species have never been confirmed to co-occur in North American ecosystems. We examined 1760 earthworms collected from 30 sites across northeastern USA, and identified them using a new morphological key. Our data show that sympatric occurrence of at least two, and often all three, species is more common than having only one species. In addition, A. tokioensis was dominant in many of these earthworm communities. The status change in species composition from only one species to two or three and the shift in dominance are most likely caused by previous incorrect species identification. Our results support expansion of known range of A. tokioensis and M. hilgendorfi northward and westward into states with colder winters. This range expansion may have taken place alongside that of A. agrestis in the last 10–20 years, but has long been overlooked. Altogether, results highlight an urgent need for correct species identification. The recognition of an expanding multi-species system represents a unique opportunity to further evaluate complex interactions among co-invading and resident species, and to investigate whether interspecific interactions have unexpected non-additive impacts on ecological processes.     

Effects of human disturbance on movement,foraging and bed selection in red deer<Emphasis Type="Italic">Cervus elaphus xanthopygus</Emphasis> from the Wandashan Mountains,northeastern China

We characterized 716 sites (including 22 beds, 60 foraging and 604 movement sites) used by red deerCervus elaphus xanthopygus Milne-Edwards, 1867 in the Wandashan Mountains, northeastern China in the winter period. We used covariates for vegetation, topography, disturbances by other ungulates, and disturbance by humans to develop movement, forage and bed site resource-selection models. We used an information-theoretic approach to select the top 5 models for movement, forage and bed site occurrences respectively. The three most parsimonious autologistic models were good predictors of movement, forage and bed occurrence of this species. Vegetation covariates were important components of all models. We recorded avoidance responses by the behaviours of species to each disturbance type (ie, villages, forest roads and abandoned roads); bed site occurrences were most sensitive to all human disturbances for their lowest odds encountered. Across all parsimonious models, villages have the largest negative effect on movement, foraging and bed occurrences. Movement and bedding behaviours were also affected by other ungulate disturbances. Altitude had slightly negative effect on movement and foraging behaviours. Ridges, topographic aspect and shrub stands were all correlated with bed-site selection. Although different behaviours may be associated with different microhabitat features, behaviours of red deer responded similarly to the same human disturbance on a broad scale. Based on the observations above, we believe that red deer are avoiding human-altered habitat for movement, bedding and foraging because of disturbances during the critical late winter period. Furthermore, various behavioural resource selection models and corresponding graphs of important habitat disturbances can be used to guide and evaluate future development proposals.     

Description and epidemiological implications of <Emphasis Type="Italic">S. scabiei</Emphasis> infection in roe deer (<Emphasis Type="Italic">Capreolus capreolus</Emphasis>) originating from chamois (<Emphasis Type="Italic">Rupicapra rupicapra</Emphasis>)

The authors describe a case of Sarcoptes scabiei transmission from a chamois (Rupicapra rupicapra) kid to three roe deer (Capreolus capreolus) fawns kept in captive cohabitation in a wildlife rehabilitation centre for approximately 4 months. Symptoms in the two male fawns started developing 1 week after skin lesions were noticed in the kid, whereas a delayed (75 days later) appearance of scabies signs was observed in the female fawn. A severe generalised form of scabies rapidly developed in a male fawn, but lesions in the other two healed after triple inoculation of ivermectin at 15 days intervals. These roe deer were monitored for specific anti-S. scabiei antibodies with an immunoblotting technique. The strongest antibody response was exhibited during the clinical phase of the infection and then rapidly declined post-treatment. Interestingly, the female fawn developed immunoglobulin G to S. scabiei proteins more than 2 months before the onset of clinical scabies. Since all captive roe deer appeared sensitive to infection with S. scabiei var. rupicaprae, the authors hypothesise that the sporadic frequency of spontaneous scabies in this Cervid host may be attributed to avoidance of direct contacts with moribund or recently dead scabietic chamois, and/or to a self-limiting (and therefore difficult to detect) course of the disease after infrequent infectious contacts and/or exposure to lower infectious doses than in the described episode.     

Secondary production,calcification and CO<Subscript>2</Subscript> fluxes in the cirripedes <Emphasis Type="Italic">Chthamalus montagui</Emphasis> and <Emphasis Type="Italic">Elminius modestus</Emphasis>

Calcification, a process common to numerous marine taxa, has traditionally been considered to be a significant source of CO₂ in tropical waters only. A number of relatively recent studies, however, have shown that significant amounts of CO₂ are also produced in temperate waters, although none of these studies was carried out on rocky shores, which are considered to be very productive systems. We compared the CO₂ fluxes due to respiration and calcification in two temperate species, the cirripedes Chthamalus montagui and Elminius modestus. The population dynamics of both species were estimated at two sites during a 1-year experimental period in order to establish mean organic (ash-free dry weight) and CaCO₃ (dry shell weight) production. Based on these parameters, we estimated the CO₂ fluxes due to respiration and calcification. CaCO₃ production was estimated to be 481.0 and at each site, representing 3.4 and respectively, of released CO₂. These fluxes represent each 47% of the CO₂ released as a result of respiration and calcification. The production of CaCO₃ at the high-density site was: (1) among the highest values obtained for temperate organisms, and (2) comparable to the estimated CO₂ fluxes for coral reefs. As calcifying organisms are well represented in temperate ecosystems in terms of both density and biomass, our results provide clear evidence that calcification of temperate organisms should not be underestimated. Additional studies on other rocky shore taxa are needed before the relative importance of calcification in rocky intertidal carbon budgets can be generalized.     

Long-term changes in lowland calcareous grassland plots using <Emphasis Type="Italic">Tephroseris integrifolia</Emphasis> subsp. <Emphasis Type="Italic">integrifolia</Emphasis> as an indicator species

We investigated the changes to calcareous grassland plots within protected sites, and whether Tephroseris integrifolia subsp. integrifolia can act as a useful indicator species for re-visitation studies within vegetation predicted to remain relatively stable. Twenty-two plots located across lowland England and all formerly containing T. integrifolia were re-surveyed following the methodology used in the original survey undertaken in the 1960s. Pseudo-turnover and between-observer bias were minimised by sampling replicate quadrats at each fixed plot using a single surveyor and at a similar time of year as the original survey. Qualitative details concerning grazing management were obtained for all sites. In contrast to other long-term re-visitation studies, all our study plots were intact and retained diverse, herb-rich vegetation, demonstrating the value of site protection. However, there were clear shifts in vegetation composition, most notably where T. integrifolia was absent, as shown by an increase in Ellenberg fertility and moisture signifying nutrient enrichment, and a decrease in the cover of low-growing, light-demanding specialists, with a change likely to be associated predominantly with grazing management. Whereas in the mid-20th century the greatest threat to calcareous grassland was habitat loss, undergrazing or temporary neglect now appears to pose the principal threat. Distinctive species such as T. integrifolia with marked sensitivity to habitat change provide a potentially useful tool for rapid assessment and monitoring of site quality. Focusing monitoring on such species allows non-expert observers to recognise the early stages of habitat degradation, providing, in effect, a “health check” on individual sites and groups of sites.     

<Emphasis Type="Italic">Ty</Emphasis>1<Emphasis Type="Italic">/copia</Emphasis>- and <Emphasis Type="Italic">Ty</Emphasis>3<Emphasis Type="Italic">/gypsy</Emphasis>-like DNA sequences in <Emphasis Type="Italic">Helianthus </Emphasis>species

Two repeated DNA sequences isolated from a partial genomic DNA library of Helianthus annuus, p HaS13 and p HaS211, were shown to represent portions of the int gene of a Ty3 /gypsy retroelement and of the RNase-Hgene of a Ty1 /copia retroelement, respectively. Southern blotting patterns obtained by hybridizing the two probes to BglII- or DraI-digested genomic DNA from different Helianthus species showed p HaS13 and p HaS211 were parts of dispersed repeats at least 8 and 7 kb in length, respectively, that were conserved in all species studied. Comparable hybridization patterns were obtained in all species with p HaS13. By contrast, the patterns obtained by hybridizing p HaS211 clearly differentiated annual species from perennials. The frequencies of p HaS13- and p HaS211-related sequences in different species were 4.3x10(4)-1.3x10(5) copies and 9.9x10(2)-8.1x10(3) copies per picogram of DNA, respectively. The frequency of p HaS13-related sequences varied widely within annual species, while no significant difference was observed among perennial species. Conversely, the frequency variation of p HaS211-related sequences was as large within annual species as within perennials. Sequences of both families were found to be dispersed along the length of all chromosomes in all species studied. However, Ty3 /gypsy-like sequences were localized preferentially at the centromeric regions, whereas Ty1/ copia-like sequences were less represented or absent around the centromeres and plentiful at the chromosome ends. These findings suggest that the two sequence families played a role in Helianthusgenome evolution and species divergence, evolved independently in the same genomic backgrounds and in annual or perennial species, and acquired different possible functions in the host genomes.