Design and evaluation of nematode 18S rDNA primers for PCR and denaturing gradient gel electrophoresis (DGGE) of soil community DNA

Consensus nematode 18S ribosomal DNA primers were designed by aligning available 18S sequences and identifying a variable region flanked by highly conserved regions. These primers were then used to amplify nematode 18S rDNA from whole soil community DNA extracted from a range of European grassland types. Cloning of the PCR amplicons (778 bp) followed by restriction digest analysis (RFLP) resulted in the recovery of 34 unique nematode sequences from the four grasslands studied. Comparison of these data with the limited number of 18S rDNA nematode sequences currently held in on-line databases revealed that all of the sequences could be assigned to known nematode taxa albeit tentatively in some cases. Two of the sequences recovered from the site in the Netherlands (wet, hay-grassland) were recovered in a clade that included a sequence of the genus Trichodorus whilst other sequences from this site showed similarity with 18S rDNA sequences of the genus Prismatolaimus (five sequences), Xiphinema (one sequence) and Enoplus (one sequence). Of the remaining sequences, two showed some affinity with Mylonchulus (UK, upland peat), four with Steinernema (UK) and one sequence with Mesorhabditis (Hungary, east European Steppe). Three sequences from the Netherlands and one from Hungary were recovered in a clade that included a sequence of the genus Pratylenchoides whilst three further sequences from the Netherlands and two from Hungary were recovered in a clade encompassing the genus Globodera. Of the remaining nine sequences, two (NL6, NL62) formed a distinct lineage within the Adenophorea with 90% bootstrap recovery in a paraphyletic clade that included sequences of Prismatolaimus and Trichodorus. Seven sequences (three from the Netherlands, three from the UK and one from Greece) were left unassigned though the tree topology suggested some relationship (58% bootstrap recovery) with the genus Cephalobus. To assess whether primers used to amplify 18S rDNA might be used to fingerprint genetic diversity in nematode communities in soil, the environmental sequence data were used to design a second set of primers carrying a GC-clamp. These primers amplified a 469 bp fragment internal to the region flanked by the primer set used to derive the nematode trees and were used to amplify 18S rDNA for subsequent analysis using denaturing gradient gel electrophoresis (DGGE). DGGE analysis of six major European grassland types revealed considerable genetic diversity between sites. However, the relationships seen with the DGGE data were inconsistent with previous studies where the same soils had been characterized with respect to functional and morphological diversity. To confirm that this second set of primers was amplifying nematode sequences, selected bands on the DGGE gels were extracted, PCR amplified and sequenced. The final alignment was 337 bases. These analyses revealed the presence of sequence signatures from the genera Paratrichodorus, Plectus, Steinernema, Globodera, Cephalobus and Pratylenchoides.     

Wing loading and habitat selection in forest beetles: Are red-listed species poorer dispersers or more habitat-specific than common congenerics?

Traits such as poor dispersal ability and high habitat specificity are thought to predispose some species to a greater risk of extinction than others. Habitat preferences and morphological features associated with dispersal ability were compared between red-listed species and common congenerics co-occurring in boreal forests in northern Sweden. Measurements of body size, wing loading and wing aspect ratio were used to compare dispersal abilities, while catches from experimental treatments of dead wood were used to compare habitat preferences. We also compared how restricted red-listed species were to particular sites or habitats relative to common species. The red-listed Epuraea longipennis was longer and wider, while Epuraea. deubeli weighed less than common Epuraea species. In contrast to expectations, these red-listed species had a larger wing area (relative to their body mass) and thus a lower wing-loading than congeneric species, suggesting superior dispersal abilities. The red-listed Tachinus elegans possessed intermediate morphological characteristics, compared with common congenerics. However, the relationships between the risk of extinction in fragmented habitats and size and dispersal ability are likely to be scale-dependent, so intermediate or superior dispersal abilities may increase extinction risk at some scales. Red-listed species were not found in fewer sites or habitat types than congenerics so were not more likely to be habitat specialists. However, some red-listed species preferred deeply shaded and Fomitopsis pinicola-inoculated logs, relative to congenerics, suggesting that specificity to these particular microhabitats may be connected with extinction risk.     

Cytomolecular characterization of rDNA distribution in various Citrullus species using fluorescent in situ hybridization

The 18S–28S and 5S rDNA sites are useful chromosome landmarks and provide valuable evidence about genome organization and evolution. This investigation was the first attempt to study the dynamics, distribution and directionality of rDNA gains and losses, as well as to understand the contribution of site number variation in the speciation of the genus Citrullus. In this study, we employed fluorescent in situ hybridization (FISH), using the18S–28S and 5S rDNA gene loci, to evaluate the differences between the (1) cultivated type watermelon C. lanatus var. lanatus (sweet watermelon), (2) the “bitter” desert watermelon C. colocynthis (colocynth) that is indigenous to the deserts of northern Africa, the Middle East and Asia, (3) the C. lanatus var. citroides (citron) “Tsamma” or “cow watermelon” that is known as and is indigenous to southern Africa, (4) and C. rehmii that thrive in the Namibian Desert. The FISH analyses showed that the sweet watermelon and colocynth have similar rDNA configuration. The sweet watermelon and colocynth genomes contain two 18S–28S rDNA gene loci, each located on a different chromosome, and one 5S rDNA locus which is co-localized with one of the 18S–28S rDNA gene loci. On the other hand, the C. rehmii has one 18S–28S rDNA locus and one 5S rDNA locus positioned on different chromosomes, while the citron has one18S–28S rDNA and two 5S rDNA loci, each located on a different chromosome. A FISH analysis of F₁ (citron × sweet watermelon) chromosome spreads revealed uniparental homeologous rDNA gene copies pertaining to the sweet watermelon versus the citron chromosomes, with the sweet watermelon chromosome containing the 18S–28S and 5S rDNA locus versus the citron homologue chromosome that has the 5S rDNA locus, but not the 18S–28S rDNA locus. Genomic in situ hybridization (GISH) analysis, using the entire citron genome as a probe to be differentially hybridized on sweet watermelon chromosome spreads, revealed that the citron genomic probes mainly hybridize to subtelomeric and pericentromeric regions of the sweet watermelon chromosomes, suggesting extensive divergence between the citron and sweet watermelon genomes. The FISH and GISH cytogenetic analysis here indicate major differences in genome organization between the cultivated watermelon type sweet watermelon and its counterpart citron that thrive in southern Africa and considered a useful germplasm source for enhancing disease and pest resistance in watermelon cultivars.     

Spatiotemporal variation of breeding gull species in a boreal lake complex in Finland: Implications for conservation

Understanding spatial and temporal patterns of species is a prerequisite for successful species and habitat conservation. Spatial variation in breeding sites of four gull species was studied in southern Finland in an oligo-mesotrophic lake complex covering almost 50 km² of water areas and 290 km of shoreline in three census periods in 1986-2004. Two of the species have declined and are regarded as red-listed in Finland (black-headed gull ridibundus and lesser black-backed gull L. f. fuscus) and two have increased (common gull L. canus and herring gull L. argentatus) in numbers during the past decades. The numbers of breeding pairs and the percentage similarity in the spatial distribution of pairs of each species in grid squares were compared between different census periods at resolutions of 0.25, 1 and 4 km². The common gull showed very high percentage similarity between the different census periods and consequently low spatial turnover in nesting sites, whereas the red-listed species, particularly the black-headed gull, had much higher spatial turnover. The spatiotemporal dynamics of gull species should thus be taken into account in conservation planning. If site protection is based only on information of breeding gulls in one year, a large or even major proportion of the breeding red-listed gulls might be outside the protected areas after a few decades. Due to the large spatiotemporal variation of red-listed gulls, areas to be protected should cover a rather large proportion of a boreal lake, not only individual islets or islands.     

Isolation and characterization of phosphate solubilizing bacteria from the rhizosphere of crop plants of Korea

Whole-cell fatty acids methyl ester (FAME) profile and 16S rDNA sequence analysis were employed to isolate and identify the bacterial groups that actively solubilized phosphates in vitro from rhizosphere soil of various crops of Korea. Out of several hundred colonies that grew on Pikovskaya's medium 13 best isolates were selected based on the solubilization of insoluble phosphates in liquid culture and further characterized and identified. They were clustered under the genera Enterobacter, Pantoea and Klebsiella and the sequences of three representative strains were deposited in the GenBank nucleotide sequence data library under the accession numbers AY335552, AY335553, AY335554.     

Isolation and identification of the most efficient plant growth-promoting bacteria associated with canola (Brassica napus)

 Thirteen of the most abundant Gram-negative bacteria which are able to grow in N-depleted culture conditions were isolated from the rhizoplane and endorhizosphere of canola (Brassica napus) and identified by 16S rDNA sequence analysis. Eight of these bacteria induced a significant increase in root dry weight ranging from 11 to 52%. Phylogenetic positioning based on 16S rDNA sequences indicated that at least four genera are represented, Pseudomonas, Variovorax, Agrobacterium and Phyllobacterium. The most important direct plant growth-promoting-rhizobacteria effect was found with both isolates belonging to the Phyllobacterium. Received: 18 April 2000     

甘肃本地产当归、黄芪和大黄的rDNA ITS序列分析

本研究采用改良CTAB法分别提取18份甘肃本地产当归、黄芪和大黄基因组DNA,并用PCR分别扩增其ITS1-5.8S-ITS2序列、直接测序并作序列同源性比对分析。双向测序分析结果表明,甘肃6个不同产地当归rDNA的ITS1、5.8S和ITS2序列一致,片段长度分别为215bp、162bp和223bp;供试的黄芪ITS1、5.8S和ITS2序列分别为228bp、164bp和210bp;大黄ITS1、5.8S和ITS2序列分别为160bp、159bp和164bp。供试材料的ITS1-5.8S-ITS2核苷酸序列已提交GenBank。本研究为提供甘肃当归、黄芪和大黄指纹图谱鉴别的分子标记、其道地性药材的分子鉴定和品质评价提供参考。     

Metal stress selects for bacterial ARDRA-types with a reduced catabolic versatility

In a previous study, 76 out of 899 bacteria, isolated from uncontaminated and adjacent metal-contaminated experimental plots, were found to fall within 12 clusters on the basis of a fourfold ‘amplified ribosomal DNA restriction analysis’ (ARDRA). Eight of these 12 ARDRA-clusters were re-investigated in the present study by sequencing about 500 bp of the 16S rDNA of the pertaining 51 isolates. Sequencing confirmed that, with the stringent fourfold ARDRA applied, all isolates of a given ARDRA-cluster did indeed have identical 16S rDNA. Seven ARDRA-clusters had 16S rDNA similarities of 98% or more with reference species suggesting identification was at least at the genus level. With one ARDRA-cluster, having a similarity of only 93% with Sphingobacterium heparium, identification was less certain. An unequivocal assignment of the eight ARDRA-clusters to distinct species was not possible. However, similarities >99.3% of five ARDRA-clusters with five reference species from two genera suggested identification was eventually even at the species level.Isolates of seven ARDRA-clusters from increasingly metal-contaminated soil samples were decreasingly versatile with respect to aromatic acids they could utilise as sole sources of carbon and energy. Only in one ARDRA-cluster exhibiting 99.8% similarity with Arthrobacter nicotinovorans was this effect less pronounced. The data strongly suggest that the decrease in catabolic versatility observed with metal-affected bacterial communities also applies to the genus level, if not to the species level.     

Investigation of genome polymorphism and seed coat anatomy of species of section Adenolinum from the genus Linum

Twelve species of section Adenolinum from genus Linum have been studied using cytogenetic methods (DAPI/C-banding, Ag-NOR staining, FISH with 5S and 26S rDNA probes), RAPD analysis, and seed surface ultrastructure and seed coat anatomy investigation. Karyotype analysis revealed that these species have a chromosome number of 2n = 18 or 2n = 36. The DAPI/C-banding patterns enabled the identification of all chromosomes and revealed the similarities in chromosome patterns in all of the studied species. It was found that 18-chromosome plants are diploids, whereas 36-chromosome forms are tetraploids. In diploid species, 5S and 26S rRNA genes are located in similar positions on a single pair of chromosomes or on two chromosome pairs in tetraploids. Ag-NOR staining showed NOR activity in both genomes of tetraploids. RAPD analysis and seed coat anatomy investigation revealed some interspecies differences. Species from nearby areas have more similar RAPD characteristics compared with more geographically distant species. The results of this study may serve as an additional argument in favour of a monophyletic origination of the Adenolinum species and its strong isolation from other Linum species.     

浙江乐清铁皮石斛rDNA ITS序列的克隆及序列比对

为判断浙江乐清铁皮石斛（Zhejiang Dendrobium officinale）与石斛属（Dendrobium）其它物种的亲缘关系,本文提取了该石斛鲜样的DNA,利用真核生物ITS序列通用引物ITS4/ITS5进行了扩增,并将扩增产物连接转化至大肠杆菌,在对阳性克隆测序。将得到的序列与GenBank上我国石斛属12个组中34个种的rDNAITS进行了比对,并在此基础上构建了系统发育树。序列比对结果表明浙江乐清铁皮石斛的rDNAITS与黄石斛（D.tosaense）一致,其次与铁皮石斛（D.officinale）最相近,序列相似性99%。系统发育树也表明浙江乐清铁皮石斛的rDNAITS序列与黄石斛（D.tosaense）的相似程度高于其与铁皮石斛（D.officinale）的相似程度。本研究将为利用ITS序列鉴别石斛物种种属关系提供参考。     

Genome-wide analysis of plastome sequence variation and development of plastidial CAPS markers in common potato and related <Emphasis Type="Italic">Solanum</Emphasis> species

The plastome sequence of the European cultivated potato, Solanum tuberosum subsp. tuberosum (tbr, GenBank accession no. DQ386163), was compared with that of S. bulbocastanum, a wild potato relative (blb, GenBank accession no. DQ347958), in order to characterize the degree and type of variability in different genomic regions, and develop molecular markers relevant to genetics, breeding and biotechnology of potato. One hundred forty-two and 251 PICs (Potentially Informative Characters) were found in coding and non-coding sequences (NCSs), respectively. Further, while variation in coding regions was almost exclusively due to nucleotide substitutions, 25% of PICs in NCSs of tbr and blb were due to indels, most of them mononucleotide or longer tandem repeats (micro and minisatellites). Four intergenic regions were selected for further analyses in other 16 tuber-bearing Solanum species. The rps16-trnQ ^UUG gene spacer was found to be the most variable, forty-six PICs in this region distinguishing 18 haplotypes. Analysis of haplotype relationships, based on variability in the four intergenic regions, confirmed that the most primitive species from Central America were the most distant to S. tuberosum. Finally, polymorphic sites in the same regions were used to develop a set of CAPS (Cleaved Amplified Polymorphic Sequences) markers for species/cytoplasm identification in Solanum spp.     

Saproxylic beetles in natural and man-made deciduous high stumps retained for conservation

Intensive forest management in Scandinavia has decreased the amount of dead wood required by saproxylic (wood-living) organisms. To reduce this problem, some dead wood is now retained during forest operations, often in the form of man-made high stumps (ca. 4 m high). Most often these stumps are cut with a harvester, although the stumps in this study were made with explosives. The aims of this study were to determine whether such stumps of aspen (Populus tremula) and birch (Betula spp.) could be used by red-listed saproxylic beetles (Coleoptera), and to examine how the fauna of man-made high stumps differs from that of natural stumps. We also studied how tree species, sun-exposure, stage of decay and trunk diameter influenced the fauna. In 169 samples of bark from high stumps 116 saproxylic species were found, of which 21 were red-listed. Many species, including red-listed ones, were more associated with man-made stumps than with natural stumps. However, in total, more species were found in the natural than in the man-made stumps. This is probably because man-made stumps provide a more homogeneous type of wood substrate than natural ones. Among the other variables the difference between aspen and birch was the most important. We conclude that man-made high stumps are valuable habitats for many saproxylic beetle species.     

Karyotype characterization reveals an up and down of 45S and 5S rDNA sites in <Emphasis Type="Italic">Crotalaria</Emphasis> (Leguminosae-Papilionoideae) species of the section Hedriocarpae subsection Macrostachyae

The genus Crotalaria is one of the largest within the family Leguminosae-Papilionoideae, with more than 600 species. However, few karyotypes have been described. In the present paper, five species belonging to the section Hedriocarpae were studied (subsection Machrostachyae), in order to better understand chromosomal evolution in Crotalaria. The results reveals that all species presented 2n = 2x = 16 with symmetrical karyotypes, and slight differences in the chromosome morphology. A secondary constriction was identified at short arm of the pair 1. The 45S rDNA was mapped in the secondary constriction and adjacent heterochromatin (NOR-heterochromatin) and a minor site was identified in C. ochroleuca. The 5S rDNA was mapped linked to 45S rDNA at chromosome 1 short arm in all species. Additional sites for 5S rDNA were identified in C. pallida, C. striata and C. mucronata. Heterochromatin blocks around the centromeres are not CMA⁺ neither DAPI⁺. The karyotypes of the subsection Macrostachyae are characterized by an inversion at chromosome pair one in relation to previous specialized floral species analyzed. Additional sites of 45S and 5S rDNA were assumed to be a result of transposition events by different ways. The results suggest heterochromatin differentiation and the position of ribosomal genes indicates chromosomal rearrangements during evolution. Karyotype characteristics corroborate the morphological infrageneric classification.     

Contrasting levels of mitochondrial DNA variability between mites (Penthalodidae) and springtails (Hypogastruridae) from the Trans-Antarctic Mountains suggest long-term effects of glaciation and life history on substitution rates, and speciation processes

We carried out a comparative study using mtDNA (COI) for two free-living, endemic, terrestrial invertebrate taxa—the springtail Gomphiocephalus hodgsoni (Hypogastruridae) and the mite Stereotydeus mollis (Penthalodidae) collected from sites throughout Victoria Land and the Queen Maud Mountains, Antarctica. Maximum likelihood (ML) analyses revealed low levels of intraspecific substructuring for G. hodgsoni (?2% sequence divergence) with groups mostly corresponding to geographic location. Other well-defined (morphological) species within the Hypogastruridae ranged from 15% to 22% sequence divergence relative to G. hodgsoni. In contrast, ML analyses for S. mollis showed considerably higher levels of intraspecific divergence (up to 18% sequence divergence), suggesting the presence of morphologically cryptic species. Other presently described mite species were also readily discernible (14-18% sequence divergence). Within both G. hodgsoni and S. mollis there were three main groups corresponding to: (1) Taylor Valley; (2) Beaufort Island; and (3) Ross Island together with the adjacent continental sites. We suggest that a common pattern of climatic and geological history over long-term glacial habitat fragmentation have determined the similar geographic and haplotype distributions found for both species. However, a large variation in substitution rate, around eight times slower for G. hodgsoni, compared to S. mollis, may be due to their different life history strategies on the Antarctic continent.     

Community structure and functional diversity of indigenous fluorescent Pseudomonas of long-term swine compost applied maize rhizosphere

The community structure and catabolic functional diversity of indigenous rhizosphere fluorescent pseudomonad isolates recovered from un-amended soil (control treated) and following long-term (6 years) swine compost application (compost treated) were studied. Soils collected for the pot experiments were from a Taiwanese experimental farm established in 1995 to study the long-term effects of crop rotation and organic amendment. In the pot experiments, corn seed was cultured; after the elongation stage, 13 and 32 indigenous rhizosphere fluorescent pseudomonad isolates were collected from compost-treated (n=13) and control-treated (n=32) soils. All isolates were classified by 16S rDNA restriction fragment length polymorphism (RFLP) and unweighted pair-group method using arithmetic averages analyses. Most (84.4%) of the control-treated soil isolates belonged to genotype A, which corresponded to Pseudomonas putida based on 16S rDNA sequences analysis. However, only 61.5% of compost-treated soil isolates were from genotype A; 30.7% of the isolates belonged to genotype B, whose 16S rDNA profile corresponded to Stenotrophomonas maltophilia. Differing catabolic potentials of the P. putida isolates from control- and compost-treated soils were evident; isolates from control-treated soils utilized organic acids including citric acid, cis-aconitic acid, and formic acid more actively than compost-treated soil isolates. Moreover, the latter isolates more efficiently utilized carbohydrates of i-erythritol and l-rhamnose than P. putida isolates from control-treated soil, consistent with the alteration in the catabolic functions of indigenous rhizosphere P. putida by long-term compost application.     

Effects of agri-environment management for stone curlews on other biodiversity

Two-hectare unsown, tilled fallow plots put in place under agri-environment schemes for stone curlews Burhinus oedicnemus in England were surveyed for other bird species, brown hares Lepus europaeus, carabid beetles, vascular plants, butterflies and bumblebees. The results were compared with those from surveys within the crop in the same field and in neighbouring fields. This was done to test whether agri-environment management targeted at a single species also provided benefits to wider biodiversity. All groups except carabid beetles were more abundant, more likely to be recorded, or more species rich on plots than within the crop. Carabid beetle activity–density and species richness did not differ between the plot and the crop; activity–density of one functional group (predators) was lower on plots than in the crop. Six rare arable plant species were recorded, predominantly from plots, and the plots are considered to perform comparably to other agri-environment options available in England designed specifically for arable plants. One red-listed bird species of conservation concern (skylark Alauda arvensis) was more abundant on plots than in the crop, and three other red-listed species (linnet Carduelis cannabina, yellowhammer Emberiza citrinella, corn bunting E. calandra) were more likely to be recorded on plots than in the crop. A fifth (lapwing Vanellus vanellus) was almost exclusively recorded from plots, and was more likely to be recorded on plots categorised as in good condition for stone curlews. These results demonstrate that fallow plots that have been put in place for the recovery of stone curlews have considerable value for a range of other farmland biodiversity, although management for stone curlews may be detrimental to some taxa, such as carabid beetles, that are sensitive to the relatively frequent spring tillage that these plots receive.     

Speciation in the parthenogenetic oribatid mite genus Tectocepheus (Acari, Oribatida) as indicated by molecular phylogeny

Despite the lack of universal concepts for species and speciation, both sexual and asexual organisms are expected to diversify into discrete genotypic and morphological clusters. Species-rich clusters of parthenogenetic oribatid mites likely evolved in the absence of sexual reproduction. We used nucleotide sequences of the large and small rDNA genes (18S and 28S) and the coding genes for heat shock protein 82 (hsp82) and elongation factor-1 alpha (ef-1α) for phylogenetic analyses of three morphotypes of the parthenogenetic oribatid mite genus Tectocepheus. DNA sequence divergences of the different morphotypes were similar to those of sexual species in other organisms. Maximum likelihood analyses of single genes and combined data sets were largely congruent in reconstructing the phylogeny of the Tectocepheus species. The results suggest that the different morphotypes of Tectocepheus indeed evolved in the absence of sexual reproduction, and that Tectocepheus minor, Tectocepheus velatus and Tectocepheus sarekensis are best considered distinct species. Further, the results suggest that 18S rDNA, hsp82 and ef-1α are powerful markers for resolving phylogenetic relationships of oribatid mites. Saturation plots indicated that the D3-region of 28S is much more saturated than all other examined genes. This indicates that the D3-region is unsuitable for resolving ancient splits in oribatid mites.     

Isolation and characterization of a mycorrhiza helper bacterium from rhizosphere soils of poplar stands

If mycorrhizal formation could be enhanced by co-inoculation with mycorrhiza helper bacteria (MHB) which promote rapid root colonization by specific ectomycorrhizal fungi, this would be of advantage to the poplar forest industry. A number of poplar rhizobacterial strains were isolated from 11 regions of the eastern China. Four of the isolates, SY15, DZ18, HLJ4, and PY10, were characterized as MHB potential strains based on their positive effect on growth of ectomycorrhizal fungi Pisolithus tinctorius (Pers.) and Lactarius insulsus (Fr.). Under greenhouse conditions, one of the bacterial isolate, DZ18, significantly promoted the poplar trees growth and ectomycorrhizal colonization of P. tinctorius and L. insulsus on Populus deltoides Marsh. In contrast, the other three isolates SY15, HLJ4, and PY10 promoted fungal growth in vitro experiments but did not enhance ectomycorrhizal (ECM) formation in the greenhouse experiment. Therefore, it was concluded that DZ18 can be considered as an MHB strain. DZ18 was identified as Bacillus sp. based on morphological, physiological, and biochemical analyses in combination with analysis of 16S rDNA gene sequences.     

The effects of habitat connectivity on cryptogam richness in boulder metacommunity

We investigated the effects of habitat patch size and connectivity on cryptogam richness on 288 calcareous boulders. The studied boulders were found to be a particularly suitable ‘metacommunity model system’ and allowed comparison of species richness among cryptogams with differing substrate specificity, life history and red list status. The dependence of species richness on boulder surface area was modelled using Arrhenius power model, and contrasts were analysed between connected and isolated boulders. Species richness was clearly related to boulder size, but the rates of increase of species richness with increasing boulder area (z-values) were highly variable among species groups. Highest value of z (1.17) was found for species with large propagules, and values around 0.5 were obtained for calciphilous and regionally red-listed species and z = 0.42 for perennials. The lowest value of z (0.27) occurred in calcicolous species while for all species, colonists, and species with small propagules the values were 0.3-0.4. The richness of all species was 11% higher on connected than isolated boulders, while connected boulders had 31% more of both calcicolous and regionally red-listed species and 19% more of colonists. The result suggests that in the investigated system that boulder size is a crucial determinant of cryptogam richness, and their spatial configuration in terms of connectivity may further contribute to cryptogam richness. The latter result lends support to the importance of metacommunity processes for the maintenance of cryptogam richness.     

Openness in management: Hands-off vs partial cutting in conservation forests, and the response of beetles

The appropriate management of forest reserves is debated; two major alternatives are succession to ‘wild’ state, or management to produce semi-open stands. For temperate conservation stands, there are no strong experiments replicated at landscape level. In each of 22 forests rich in oaks (Quercus spp.) in Sweden, we set up a closed-canopy wild plot (1 ha), and a cutting plot (1 ha) to produce semi-open conditions, studying them before and after cutting. About 25% of the tree basal area was cut (large trees and dead wood retained) and harvested as bio-fuel, a CO₂ - neutral energy source. We examined the response of beetles and trapped 59,000 individuals (1174 species; 100 red-listed species). For both the guild of herbivorous beetles (222 species) and of saproxylic beetles connected to oak wood (267 species), species richness increased by about 35% in the harvested plots, relative to the wild reference plots. Species composition within the groups changed, though not strongly. Thirteen saproxylic species of 50 analysed, and three herbivores of 12 increased in cutting plots. For red-listed saproxylic beetles, species richness did not change significantly. Regression analyses suggest that more open cutting plots disfavour the red-listed beetles of this forest type. Thus, partial cutting increased species diversity of two beetle groups, probably due to changed microclimate and increase in herbaceous plants, but some red-listed saproxylic beetles may be disfavoured. A hands-off alternative may through storms and other disturbances produce open patches, more dead wood, and favour some species. Combinations of these alternatives, carefully planned at the landscape level, need to be considered.