Programmed cell death during development of cowpea (Vigna unguiculata (L.) Walp.) seed coat

The seed coat develops primarily from maternal tissues and comprises multiple cell layers at maturity, providing a metabolically dynamic interface between the developing embryo and the environment during embryogenesis, dormancy and germination of seeds. Seed coat development involves dramatic cellular changes, and the aim of this research was to investigate the role of programmed cell death (PCD) events during the development of seed coats of cowpea [Vigna unguiculata (L.) Walp.]. We demonstrate that cells of the developing cowpea seed coats undergo a programme of autolytic cell death, detected as cellular morphological changes in nuclei, mitochondria, chloroplasts and vacuoles, DNA fragmentation and oligonucleosome accumulation in the cytoplasm, and loss of membrane viability. We show for the first time that classes 6 and 8 caspase‐like enzymes are active during seed coat development, and that these activities may be compartmentalized by translocation between vacuoles and cytoplasm during PCD events.     

Effects of nutrients and light on periphyton biomass and nitrogen uptake in Mediterranean streams with contrasting land uses

1. Nutrient diffusing substrata (NDS) were used to determine the relative importance of nutrients and light as potential limiting factors of periphyton biomass and nitrogen (N) uptake in Mediterranean streams subjected to different human impacts. The nutrients examined were phosphorus (P) and N, and we also further differentiated between the response of periphyton communities to N species (i.e. NO₃‐N and NH₄‐N). To examine the effect of light and nutrients on periphyton biomass, chlorophyll a accrual rates on NDS located at open and closed canopy sites were compared. The effect of nutrient availability on periphyton uptake was measured by ¹⁵N changes on the NDS after NO₃‐¹⁵N short‐term nutrient additions. 2. Results show that light was the main factor affecting algal biomass in the study streams. Algal biomass was in general higher at open than at closed canopy sites. Nutrient availability, as simulated with the NDS experiments, did not enhance algal biomass accrual in either of the 2 light conditions. 3. In the control treatments (i.e. ambient concentrations), periphyton NO₃‐N uptake rates increased and C : N molar ratios decreased consistently with increases in N availability across streams. NO₃‐N uptake rates were altered when ambient N concentrations were increased artificially in the N amended NDS. Periphyton assemblages growing on N enriched substrata seemed to preferentially take up N diffusing from the substratum rather than N from the water column. This response differed among streams, and depended on ambient N availability. 4. Periphyton biomass was not significantly different between substrata exposed to the two forms of available N sources. Nonetheless, we found differences in the effects of both N sources on the uptake of N from the water column. NH₄‐N seemed to be the preferred source of N for periphyton growing on NDS. 5. Results suggest that the effect of riparian zones on light availability, although seldom considered by water managers, may be more important than nutrients in controlling eutrophication effects derived from human activities. Finally, our results confirm that not only increases in concentration, but also stoichiometric imbalances should be considered when examining N retention in human altered streams.     

Assessment of sampling approaches for a multi‐taxa invertebrate survey in a South African savanna‐mosaic ecosystem

Invertebrate diversity is seldom included in conservation assessments, primarily because information is lacking. Broad surveys may be too costly, difficult or ineffective. Here we assess a ‘shopping basket’ approach, targeting 17 taxa using a range of methods. We sampled 43 one‐hectare sites stratified within 560 km² of heterogenous African savanna. We achieved up to 80% sampling completeness for epigaeic fauna, but generally much lower completeness (around 50%) for plant‐dwelling and flying taxa. For the former we identified duplication of methods, and for the latter, addition of methods and increased temporal variation rather than effort would improve completeness. Within a taxon, sampling 75% of species present required, on average, about 784 individuals. When considering the local richness, 75% completeness required about 27 individuals per species, but these figures require validation in other areas. About 58 sites were required to achieve 75% sampling completeness, translating to about one site per 10 km². The percentage of species sampled only in a particular month ranged between 4% and 46%, with greater temporal effects recorded for flying taxa than for epigaeic ones. The trend was similar for species unique to a particular year, with the most extreme case being 67% of the butterfly species sampled one year not previously recorded. We demonstrated and evaluated the feasibility of a simultaneous multi‐taxon survey approach to produce data useful for conservation planning and monitoring. We strongly recommend a quantified approach for surveys and inventories, with details such as specific methods decided based on the biome sampled, and taxonomic expertise available for identification.     

Pliocene and Pleistocene events shaping the genetic diversity within the central corridor of the Brazilian Atlantic Forest

Dinoponera lucida (Formicidae; Ponerinae) is an extinction‐threatened species of ant which is endemic in the central corridor of the Atlantic Forest. We used mitochondrial sequences of the Cox1, Cox2 and Cytb genes in order to infer some aspects of the evolutionary history and phylogeography of this ant. High genetic divergence and population structure were observed for the whole species. The current pattern of D. lucida diversity seems to be shaped during different geological times: middle Pliocene, early Pleistocene and mainly late Pleistocene, when the reduction of populations generated a structure pattern of the genetic variation of this species. Our data show that this structure results from the maintenance of populations of D. lucida within very small putative refuges to the south of the central Bahia refugium. We thus argue that, for some Atlantic forest endemic species, especially those resistant to very small fragments of forest, such as D. lucida, the small putative refuges were as important as, or even more important than, larger and stable refuges for the creation and maintenance of diversity, adding another piece to the puzzle of the mechanisms underlying local endemism. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 101 , 949–960.     

Unravelling evolutionary lineages among South African velvet worms (Onychophora: Peripatopsis) provides evidence for widespread cryptic speciation

The endemic South African velvet worm genus Peripatopsis currently contains eight recognized species described from variable morphological characters and the current taxonomy is unsatisfactory. In an attempt to investigate evolutionary relationships within Peripatopsis, we collected 137 individuals from 34 sample localities for six of the eight species. Sequence data derived from two partial mitochondrial (mt)DNA gene loci (COI and 12S rRNA), as well as partial sequence data from the ribosomal nuclear 18S rDNA locus in combination with gross morphological characters and scanning electron microscopy (SEM), was used to examine evolutionary relationships. Phylogenetic relationships were investigated using minimum evolution (ME) and Bayesian inferences (BI). Additionally, we also undertook a maximum likelihood (ML) analyses on the combined DNA sequence data set. The combined DNA evidence topologies derived from the ME, BI, and ML was highly congruent and was characterized by the presence of multiple lineages within recognized taxa. Peripatopsis clavigera, Peripatopsis moseleyi, and Peripatopsis sedgwicki each comprised two evolutionary lineages; Peripatopsis capensis comprised three; and Peripatopsis balfouri comprised six operational taxonomic units respectively. Genealogical exclusivity at both mtDNA and nuclear DNA among the geographically coherent groups coupled with pronounced sequence divergence suggested a two‐fold increase in the number of species within Peripatopsis. Previously used gross morphological characters (such as the number of leg pairs and colour) were either highly variable within operational taxonomic units, or were invariant, suggesting that alternative morphological characters are necessary for species discrimination. SEM results revealed potentially useful diagnostic characters that can discriminate between at least discriminate some of the newly‐identified lineages. © 2009 The Linnean Society of London, Biological Journal of the Linnean Society 2009, 97 , 200–216.