New chromosome numbers, meiotic behaviour and pollen fertility in American taxa of Lupinus (Leguminosae): contributions to taxonomic and evolutionary studies

Original chromosome determinations are presented for 20 American Lupinus taxa, including, for the first time, unifoliolate species, together with first data on meiotic behaviour and pollen fertility for some South American species. Most of the Brazilian multifoliolate L. lanatus, L. rubriflorus, L. multiflorus, L. paranensis, L. bracteolaris and L. reitzii and unifoliolate L. crotalarioides, L. guaraniticus and L. velutinus accessions analysed presented regular chromosome pairing. Meiotic indexes and estimations of pollen viability were higher than 90% for all species and accessions analysed, reflecting the generally regular meiotic behaviour of these plants. Chromosome numbers were determined for the first time for the eastern South‐American species L. guaraniticus, L. crotalarioides, L. paranensis, L. paraguariensis and L. velutinus (n = 18 or 2n = 36) and for the Andean L. ballianus, L. eanophyllus, L. huaronensis, L. semperflorens, plus another eight taxa (2n = 48) from Peru and Bolivia, and L. bandelierae (2n = 36) from Bolivia. Chromosome numbers were confirmed for L. lanatus, L. rubriflorus (2n = 36), L. bracteolaris (2n = 34) and L. microphyllus (2n = 48). In the three accessions of the North American unifoliolate species, L. cumulicola and L. villosus, a chromosome number (2n = 52) previously unknown among American taxa was found. The results of the study, plus published data, support the suggestions that south‐eastern South American species are a group cytologically differentiated from the Andean as well as from most other American ones, and that the Brazilian and the North American unifoliolate Lupinus had independent origins. © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society, 2006, 150 , 229–240.     

Response of plant species richness and primary productivity in shrublands along a north–south gradient in Europe to seven years of experimental warming and drought: reductions in primary productivity in the heat and drought year of 2003

We used a nonintrusive field experiment carried out at six sites – Wales (UK), Denmark (DK), the Netherlands (NL), Hungary (HU), Sardinia (Italy – IT), and Catalonia (Spain – SP) – along a climatic and latitudinal gradient to examine the response of plant species richness and primary productivity to warming and drought in shrubland ecosystems. The warming treatment raised the plot daily temperature by ca. 1 °C, while the drought treatment led to a reduction in soil moisture at the peak of the growing season that ranged from 26% at the SP site to 82% in the NL site. During the 7 years the experiment lasted (1999–2005), we used the pin‐point method to measure the species composition of plant communities and plant biomass, litterfall, and shoot growth of the dominant plant species at each site. A significantly lower increase in the number of species pin‐pointed per transect was found in the drought plots at the SP site, where the plant community was still in a process of recovering from a forest fire in 1994. No changes in species richness were found at the other sites, which were at a more mature and stable state of succession and, thus less liable to recruitment of new species. The relationship between annual biomass accumulation and temperature of the growing season was positive at the coldest site and negative at the warmest site. The warming treatment tended to increase the aboveground net primary productivity (ANPP) at the northern sites. The relationship between annual biomass accumulation and soil moisture during the growing season was not significant at the wettest sites, but was positive at the driest sites. The drought treatment tended to reduce the ANPP in the NL, HU, IT, and SP sites. The responses to warming were very strongly related to the Gaussen aridity index (stronger responses the lower the aridity), whereas the responses to drought were not. Changes in the annual aboveground biomass accumulation, litterfall, and, thus, the ANPP, mirrored the interannual variation in climate conditions: the most outstanding change was a decrease in biomass accumulation and an increase in litterfall at most sites during the abnormally hot year of 2003. Species richness also tended to decrease in 2003 at all sites except the cold and wet UK site. Species‐specific responses to warming were found in shoot growth: at the SP site, Globularia alypum was not affected, while the other dominant species, Erica multiflora, grew 30% more; at the UK site, Calluna vulgaris tended to grow more in the warming plots, while Empetrum nigrum tended to grow less. Drought treatment decreased plant growth in several studied species, although there were some species such as Pinus halepensis at the SP site or C. vulgaris at the UK site that were not affected. The magnitude of responses to warming and drought thus depended greatly on the differences between sites, years, and species and these multiple plant responses may be expected to have consequences at ecosystem and community level. Decreases in biodiversity and the increase in E. multiflora growth at the SP site as a response to warming challenge the assumption that sensitivity to warming may be less well developed at more southerly latitudes; likewise, the fact that one of the studied shrublands presented negative ANPP as a response to the 2003 heat wave also challenges the hypothesis that future climate warming will lead to an enhancement of plant growth and carbon sequestration in temperate ecosystems. Extreme events may thus change the general trend of increased productivity in response to warming in the colder sites.     

The taxonomic and evolutionary significance of some juvenile characters in the Brassiceae

The shape of the cotyledon lamina and a few other juvenile characters are studied for some 140 taxa of the cruciferous tribe Brassiceae. They cast a number of doubts on the present botanical status of certain taxa, such as Diplotaxis siifolia, Erucastrum cardaminoides and Brassica balearica. The whole tribe Raphaninae appears rather unnatural when viewed from cotyledon evidence, which suggests that beak development and/or fruit reduction have evolved on different occasions. A primitive cotyledon type seems to be that showed by the genus Sinapidendron. It is nowadays still present in Brassica balearica, Oudneya africana and the oriental genus Orychophragmus. Some phenetic lines now represented by Moricandia, Foleyola, Hemicrambe, Diplotaxis and Brassica repanda may indicate ancient evolutionary trends derived from the primitive Sinapidendron-like Miocenic stock. Brassica ancestors could have either resembled B. repanda or evolved along the phenetic line now represented by the Diplotaxis Erucastrum-Brassica complex. A diphyletic or polyphyletic origin for the genus Brassica becomes thinkable.     

Photolytic Decarboxylation of Carboxyl-14C-labelled Indol-3yl-acetic Acid in Leaves of the Apple Tree

The nature and rate of degradation of carboxyl-¹⁴C-labelledindol-3y1-acetic acid (IAA-[l-¹⁴C]) were studied in apple leaves.The labelled auxin was applied to the cut surface of the growingshoot after the apical part had been removed. The respiratoryCO₂ absorbed by chromatographic paper as Na₂CO₃ then freed byphosphoric acid was quantitatively measured by an internal gascounter. It was found that the concentration of ¹⁴CO₂ evolvedby leaves was 77 times higher in daylight than in darkness.The ratio of ¹⁴CO₂/CO₂ obtained from respiration from the uppersurface of leaf blades was two and seven times higher than thatfrom the lower surface after 15 and 30 h of daylight, respectively.No such differences were noticed in darkness. Similarly, thetotal radioactivity of leaf tissues tripled in daylight, presumablybecause of photosynthetic incorporation of radioactive CO₂ evolvedduring decomposition of LAA. These facts demonstrate the photolyticcharacter of auxin decarboxylation in apple leaves. Prolongeddarkness seemed to provoke a large metabolite withdrawal fromleaves and, to some extent, to protect auxin against decarboxylation.     

A Simple Method for Paraffin and Plastic Embedding of Protozoa

SUMMARY. To avoid multiple centrifugation and considerable losses of material in preparing protozoa for paraffin and plastic embedding a simple method has been worked out, requiring only two centrifugations, one before and one after fixation. During the second centrifugation, which is done at 700 g for 10 minutes, the organisms form a cohesive pellet which may be removed from the centrifuge tube by means of a fine spatula and handled as a piece of tissue through the whole process of dehydration and embedding.     

A Novel Healing Filament in Ciliate Regeneration

The interphase cells of the hypotrich ciliate Paraurostyla weissei possess a complex fibrillar system surrounding basal bodies in the compound ciliary assemblages, cirri and membranelles. During replacement of the ciliature at cell division, transient filaments precede and accompany the development of ciliary primordia and participate in the formation of the fission furrow. Both fibrillar systems are recognized by monoclonal antibody FXXXIX 12G9. We studied regeneration of cellular fragments after transection employing the mAb 12G9 and found a new cytoskeletal structure involved in healing of the excisional wound. The healing filament is formed at the wound edge, distally and in connection with the bases of cirri closest to the wound. It is visible 5 min after transection. Concomitant with development of new ciliary primordia, the healing filament shrinks and finally disappears together with other transient fibers formed in this process. Ultrastructural analysis of immunolabeled regenerating cells revealed that structures recognized by mAb 12G9 contain fine filaments whose packing and arrangement depends on accompanying cytoplasmic elements and the developmental status of a fragment. Assembly of the healing fiber does not depend on microtubules and microfilaments since it develops in cellular fragments exposed to cold, nocodazole, and Cytochalasin D. On Western blots of whole cell and cytoskeletal extracts of P. weissei the 12G9 antibody identified one protein band whose molecular weight corresponds to 60 kDa.     

FORMULATING A NEW PASSION FRUIT JUICE BEVERAGE WITH DIFFERENT SWEETENER SYSTEMS

The aim of this work was to optimize the acceptability of a natural passion fruit beverage using different levels of passion fruit pulp and sucrose, and to determine the equi-sweet concentrations of aspartame, sucralose, and a blend of 80% aspartame  +  20% acesulfame-K for the optimized formula. A 2² central composite design was used to optimize the acceptability of the sucrose-sweetened beverage, which was accessed using a 9-point structured hedonic scale. Acceptability data were fitted to a second-order model equation provided in the design. The selected pulp content and sucrose concentration were, respectively, 2.5 ° Brix and 10%. Measurements of sweetness equivalence were accomplished using two types of sensory methods: magnitude estimation and difference-from-control tests. The concentrations of aspartame, sucralose and a blend of 80% aspartame  +  20% acesulfame-K found as equi-sweet to 10% sucrose in the studied passion fruit beverage were, respectively, 0.043, 0.016 and 0.026%.

PRACTICAL APPLICATIONS

This article deals with formulating a new passion fruit juice beverage with different sweetener systems. The research makes a very important contribution to the literature on sensory science used on product development by demonstrating the necessity to study the substitution of sucrose by high-intensity sweeteners every time a formulation is changed or a new product is developed.     

Population genomics of the inbred Scandinavian wolf

The Scandinavian wolf population represents one of the genetically most well-characterized examples of a severely bottlenecked natural population (with only two founders), and of how the addition of new genetic material (one immigrant) can at least temporarily provide a 'genetic rescue'. However, inbreeding depression has been observed in this population and in the absence of additional immigrants, its long-term viability is questioned. To study the effects of inbreeding and selection on genomic diversity, we performed a genomic scan with approximately 250 microsatellite markers distributed across all autosomes and the X chromosome. We found linkage disequilibrium (LD) that extended up to distances of 50 Mb, exceeding that of most outbreeding species studied thus far. LD was particularly pronounced on the X chromosome. Overall levels of observed genomic heterozygosity did not deviate significantly from simulations based on known population history, giving no support for a general selection for heterozygotes. However, we found evidence supporting balancing selection at a number of loci and also evidence suggesting directional selection at other loci. For markers on chromosome 23, the signal of selection was particularly strong, indicating that purifying selection against deleterious alleles may have occurred even in this very small population. These data suggest that population genomics allows the exploration of the effects of neutral and non-neutral evolution on a finer scale than what has previously been possible.