Differential abundance of simple repetitive sequences in species ofBrassica and relatedBrassicaceae

SixBrassica species, known as the triangle of U, and four species from related genera were characterized by DNA fingerprinting with simple repetitive oligonucleotide probes. Our results show that CT-, TCC-, and GTG-repeat motifs are equally abundant in the genomes of the sixBrassica species. In contrast, GATA-, GGAT-, and GACA-multimers are unevenly distributed among different species. As judged from the number and strength of hybridization signals, the highest copy number of all three motifs occurs inBrassica nigra, while the lowest is observed inB. oleracea. The abundance of GATA-and GACA-repeats varies in a coordinate way. The amphidiploid genomes ofB. juncea, B. carinata, andB. napus each harbour intermediate amounts of (GATA)₄ and (GACA)₄-detected repeats as compared to their diploid progenitors, thus supporting the concept of the U triangle. GATA-, GACA-, and GGAT-repeats were also abundant inEruca sativa andSinapis arvensis, but not inRaphanus sativus andSinapis alba. These results support the idea thatBrassica nigra is more closely related toSinapis arvensis than to otherBrassica species such asB. rapa andB. oleracea.     

Host size and sex allocation of parasitoids in a gall forming community

Summary The sex ratios of three Mesopolobus species were found to vary in different hosts in a gall-forming community in Berkshire. These relationships were consistent with Charnov's host size—sex ratio hypothesis (Charnov 1979). Due to the segregation of host species on different species of Quercus, it was possible to conduct a geographical variation experiment proposed by Charnov et al. (1981). The results of this experiment suggest that the parasitoids have a fixed rule of thumb and are not flexible in their sex allocation in response to changes in host size distribution. The data, therefore, do not support the prediction of Charnov's hypothesis. Problems associated with this kind of field experiment are discussed.     

The close phylogenetic relationship of Nitrobacter and Rhodopseudomonas palustris

The phylogenetic position of Nitrobacter winogradskyi and two other nitrite-oxidizing bacteria was elucidated comparing oligonucleotides of the 16S ribosomal RNA. Nitrobacter winogradskyi and the Nitrobacter isolate Yukatan are genetically nearly identical; Nitrobacter isolate X14 is more distantly related. Phylogenetically, Nitrobacter is a member of a group of purple non-sulfur bacteria that is defined by various species of Rhodopseudomonas, Rhodomicrobium vannielii, Rhodospirillum rubum and their non-phototrophic relatives. Nitrobacter shares a high sequence similarity to Rhodopseudomonas palustris. These findings are in accord with several common taxonomic characteristics, and in addition support the conversion hypothesis for the origin of this group of chemolithotrophic bacteria.     

Branchinecta belki n. sp. (Branchiopoda: Anostraca), a new fairy shrimp from Mexico,hybridizing withB. packardi Pearse under laboratory conditions

A new fairy shrimp,Branchinecta belki n.sp., endemic to the south of Coahuila state is described and figured. A total of nine species of phyllopods, including the new species, occur in ponds in the type area.The laboratory hybridization ofB. belki andB. packardi through no-choice mating tests in reciprocal crosses is discussed. A mixture of characteristics of parental species is present in male F1 and F2 hybrids. This may provide a biological tool, or search image (sensu Wiman, 1979a), for detecting male hybrids, should such exist, between theBranchinecta species of this study in nature. In addition to the reported interspecific hybridizations inStreptocephalus (Wiman, 1979a & 1979b) and inArtemia (Bowenet al., 1985) under laboratory conditions, the new evidence inBranchinecta suggests that absence of efficient premating mechanisms may be common in Anostraca.     

The evolutionary significance of multigermicity in the genusSpinacia (Chenopodiaceae)

The presence of multigerm seedballs in the chenopodiacious genusSpinacia is noted. In the wild, colonising, and weedy dioecious species,S. turkestanica andS. tetrandra, the distribution of a multigerm seedball could effectively overcome the problem posed by isolation of the sexes. The hypothesis is tested assessing the extent of intra-seedball progeny hybridisations and seed production in the two wild species. The success in seed production by intra-seedball progeny crosses suggests that the distribution of such seedball progeny groups permits not only a percentage survival under isolation of these dioecious plants, but also the colonisation of areas outside that of the parent populations.     

Somaclonal variation within poplar

One thousand and ninety-two poplars were regenerated in vitro from callus of 13 poplar clones representing the Leuce, Aigeiros and Tacamahaca sections. At lest 44 of the regenerants differed in some way from the original clones. Somaclonal variation occurred more frequently in poplars of the Leuce section (8%) than in those of the Aigeiros or Tacamahaca sections (1%). Variation was noticed in growth habit, leaf shape or indentation but not in the reaction to four Melampsora races. However, after one growing season in the field, a few regenerants from calli of two clones (Ogy and Rap) differed in their susceptibility vis à vis the original clones. Cultivation of callus from Leuce poplars that had survived exposure to increasing concentrations of toxins from Hypoxylon mammatum gave rise to a toxin-tolerant line from which toxin tolerant plants were regenerated. Flow cytometry to measure the DNA content of nuclei showed that regenerants tended to be tetraploid.Abbreviations NAA naphthaleneacetic acid - BAP benzylaminopurine - TDZ thidiazuron - MS Murashige & Skoog medium     

Herbivory and tree mortality across a pinyon pine hybrid zone

We examined the abundances of three common insect herbivores on pure and hybrid pinyon pines along a 250-km transect in west-central Arizona, United States. Using six morphological traits, we developed a hybrid index to classify trees as pure Pinus californiarum, hybrid, or pure Pinus edulis. The insects (the stem-boring moth, Dioryctria albovittella, the scale insect, Matsucoccus acalyptus, and several species of pitch moths that produce wounds on the trunk and branches) exhibited different distributional patterns across tree types. Stem-boring moths were significantly more abundant on trees at hybrid sites compared to trees at pure sites. In addition, within hybrid sites, hybrids supported significantly more moth larvae than pure trees of either species. These two patterns support the hybrid susceptibility hypothesis in which hybrid breakdown results in increased susceptibility to herbivory. In contrast to stem-borers, there were significantly more pitch moth wounds on trees at pure P. californiarum sites than at hybrid and pure P. edulis sites. Within the hybrid zone, pitch moth abundance was equal on pure P. californiarum and hybrids, and both were significantly greater than on pure P. edulis. These within-site comparisons support the dominance hypothesis where hybrid resistance differs from one tree species, but not the other. Scale insects exhibited the most restricted distribution; over the 250 km transect they were found only in the hybrid zone. This supports the hybrid susceptibility and/or the stress hypothesis (i.e., species at the edge of their range suffer greater stress and are more susceptible to herbivory). We summed the mean numbers of these three common herbivores across sites and found that hybrid sites supported 2.1 and 3.9 times more herbivores than pure P. californiarum and P. edulis sites, respectively. Furthermore, tree mortality was on average, 35 times greater within the hybrid zone compared to pure zones of each species and was associated with the cumulative abundance of herbivores (r ²=0.646). Regardless of whether this mortality is due to insect infestation, stress or a combination of both, these results suggest that hybrid zones are important arenas of natural selection.     

The systematics of Porphyra: character evolution in closely related species

Isozymes, vegetative and reproductive morphology, seasonality, vertical and geographic distributions and chromosomes were compared for six pairs of putative sibling species of Porphyra (P. abbottae/P. torta, P. fallax subsp. fallax/P. fallax subsp. conwayae, P. amplissima/P. cuneiformis, P. fucicola/P. leucostica, P. miniata/P. variegata, P. umbilicalis/P. umbilicalis) and among five species in a complex (P. brumalis, P. kurogii, P. linearis, P. pseudolinearis, and P. purpurea.) Geographic distribution and zymograms for certain proteins showed the greatest change between species pairs: only one pair of species had identical distributions, and most species pairs were disjunct; every species had a different allozyme for GOT-1, whereas all species had apparently identical proteins for phycoerythrin. Seasonality and habitat exhibited moderate differentiation: Northeast Pacific sibling species were characterized by a high intertidal winter species pairing with a mid intertidal spring species, whereas all but one of the other species pairs exhibited nearly identical vertical distributions and seasonalities. There were few changes in morphology: most species pairs had essentially identical morphologies and coloration and the same arrangement of reproductive cells. Chromosome numbers and karyotypes were identical for species pairs and in the species complex. These results provide evidence for different rates of evolution of different characters in the genus Porphyra.     

Analysis of the Properties of <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> Insecticidal Toxins Using a Potential-sensitive Fluorescent Probe

A potential-sensitive fluorescent probe, 3,3-dipropylthiadicarbocyanine iodide, was used to analyze, at pH 7.5 and 10.5, the effects of Bacillus thuringiensis toxins on the membrane potential generated by the efflux of K⁺ ions from brush border membrane vesicles purified from the midgut of the tobacco hornworm, Manduca sexta. Fluorescence levels were strongly influenced by the pH and ionic strength of the media. Therefore, characterization of the effects of the toxins was conducted at constant pH and ionic strength. Under these conditions, the toxins had little effect on the fluorescence levels measured in the presence or absence of ionic gradients, indicating that the ionic selectivity of their pores is similar to that of the intact membrane. Valinomycin greatly increased the potential generated by the diffusion of K⁺ ions although membrane permeability to the other ions used to maintain the ionic strength constant also influenced fluorescence levels. In the presence of valinomycin, active toxins (Cry1Aa, Cry1Ab, Cry1Ac, Cry1C and Cry1E) efficiently depolarized the membrane at pH 7.5 and 10.5.     

Evaluation of random amplified polymorphic DNA for species identification and phylogenetic analysis inStylosanthes (Fabaceae)

Random amplified polymorphic DNA (RAPD) was assessed for its suitability as a tool to be used in the identification of taxa from the genusStylosanthes (Fabaceae, Papilionoideae, Aeschynomeneae). Five random primers were used to fingerprint accessions from seven species in the genus, and generated RAPD profiles that were species-specific. Data were used to examine evolutionary relationships between taxa, employing both clustering and ordination techniques, and the results were compared with those from a previous cladistic analysis of chloroplast DNA (cpDNA) restriction fragments. Both multivariate approaches indicated relationships that were generally similar to those obtained by RFLP analysis of cpDNA. However, while cluster analysis grouped together all accessions within species, ordination placed certain accessions ofS. humilis, S. macrocephala andS. capitata into separate groups. Experiments to test the assumed homology of comigrating RAPDs estimated 85.7% homology for accessions within species, and 53.8% homology for accessions between species. The value of RAPD data in systematics is discussed.     

In vitro culture of Dianthus zeyheri subsp. natalensis,a South African carnation

A South African carnation species Dianthus zeyheri subsp. natalensis was cultured in vitro using techniques similar to those developed for the cut carnation (Dianthus caryophyllus L.). The ease of callus and suspension culture establishment makes this species a useful tool for fundamental biochemical studies.Abbreviations 2,4-d 2,4-dichlorophenoxyacetic acid - NAA -naphthaleneacetic acid - KIN kinetin - pCPA p-chlorophenoxyacetic acid     

Seasonal water potential changes and proline accumulation in mediterranean shrubland species

We studied the water relations of 6 shrub and 3 tree species typical of the mediterranean climate region of central Spain to identify differential responses to water stress between and within species, and to determine if free proline concentration in leaves could be used as a water stress indicator. Predawn and midday water potentials (w) on a seasonal basis, relative water content (RWC), leaf mass per area, foliar nitrogen and free proline concentrations were measured. The lowest water potentials were observed at the end of the summer, with recovery to higher water potentials in the fall and winter seasons. Species differed regarding the annual w fluctuation. Thymus zygis, Halimium viscosum, Genista hirsuta and Juniperus oxycedrus exhibited the most negative midday and predawn w (both less than -6 MPa) with a large magnitude of response to changing conditions in soil moisture of the upper horizon of the soil. Lavandula pedunculata and Cistus ladanifer showed a moderate response. Quercus rotundifolia, Quercus faginea and Retama sphaerocarpa showed a modest response. The w of different size individuals of Quercus rotundifolia and Cistus ladanifer were compared. The annual w fluctuation was greater in small individuals as compared to large individuals. In every species, there was an increase in proline concentration of bulk leaf tissues when predawn w dropped below -5 MPa. Small plants of Cistus ladanifer reached lower water potentials and also higher concentration of proline than bigger plants. Proline could possibly be used as a drought stress indicator in every species except Q. rotundifolia. It is suggested that in addition to water stress avoidance due to deep root systems, some mechanisms of water stress tolerance may operate among shrub and tree species of central Spain.     

Ibf-1 (Iodine binding factor), a highly variable marker system in the Triticeae

Summary Isoelectric focusing of extracts from the endosperm of mature grains of hexaploid wheat and related species was used to study the genetic control of Iodine binding factor (IBF). Ten IBF bands were present in Chinese Spring (CS) and analysis of the nullisomictetrasomic and ditelosomic lines of CS showed nine of them to be controlled by genes on the long arms of the homoeologous group 5 chromosomes. Five alleles were detected at Ibf-A1 locus, four at Ibf-B1 and four at Ibf-D1 among a sample of 46 wheat genotypes. Homoeoloci were found on chromosome 5R of Secale cereale, 5E of Agropyron elongatum, 5U of Aegilops umbellulata, 5Agⁱ of Agropyron intermedium, 5S¹ and 4S¹ of Aegilops sharonensis and 4H of Hordeum vulgare.     

Seed morphology in Southern AfricanOrchidoideae (Orchidaceae)

The seed morphology of 151 species of Southern AfricanOrchidoideae (Orchideae andDiseae; sensuDressler 1981) was studied by means of scanning electron microscopy. Two different seed types were found. (1) In the majority of species the seeds are minute and fusiform. The seed coat is made up of comparatively few concave and elongate testa cells with straight or slightly undulate and generally unthickened anticlinal cell walls. The seed type was here termed Satyrium-type. While most species are very similar in the ornamentation of the periclinal walls of their seed coat, considerable variation was found inHolothrix where two distinct groups can be recognized in this respect. (2) A remarkably different seed type was observed inDisa uniflora and three apparently closely related species (Disa uniflora-type), where large balloon-like seeds occur. Their seed coat consists of convex cells with undulate anticlinal walls. It is suggested that this seed type is a derived condition and has evolved in adaptation to the specialized habitat alongside streams. The possibility of hydrochory in these four species is briefly discussed.     

Abundance and half-life of the distinct oat phytochrome A3 and A4 mRNAs

Gene-preferential oligonucleotide probes were used to determined the relative abundance and half-lives of distinct oat phytochrome A (PHYA) mRNAs. Oat PHYA mRNAs are highly conserved in the 5-untranslated region and the coding region, but the 3-untranslated region has an overall lower sequence conservation and was the source of gene-preferential probes. PHYA3 mRNA was estimated to be ca. 61% of the oat PHYA mRNA pool present in poly(A)⁺ RNA from dark-grown seedlings. The half-lives for PHYA3 and PHYA4 mRNAs were both estimated to be ca. 30 min, and a similar short half-life was estimated for the average PHYA mRNA. Sequence comparisons of PHYA mRNAs from four grass species identified conserved sequences within the 5- and 3-untranslated regions that might be important for PHYA mRNA degradation.     

Plant regeneration from callus and explants of Sesbania spp.

Root, hypocotyl and cotyledon explants of Sesbania bispinosa, Sesbania cannabina, Sesbania formosa, and Sesbania sesban were cultured on Murashige and Skoog medium with benzyladenine (BA; 2.22, 4.44, 8.88 M) in combination with 2,4-dichlorophenoxyacetic acid (2,4-d; 2.26, 4.52, 9.05 M), indolebutyric acid (IBA; 0.25, 0.49, 4.92 M) or naphthaleneacetic acid (NAA; 2.69, 5.37, 10.74 M). Although all explant types developed some callus, callus occurred earliest and continued to grow fastest with hypocotyls. Media including 2.4-d or NAA gave the fastest growing callus. Callus was subcultured up to 10 times at 20-day intervals and retained a rapid growth rate. Shoots regenerated readily from both hypocotyls or cotyledons but not from roots. Shoot organogenesis was most frequent with IBA (0.25–4.92 M) in combination with BA (4.44–8.88 M) and did not occur with 2,4-d. With each species at least one medium induced shoot differentiation from more than 50 percent of the callus pieces. With one exception, media containing IBA that induced shoot organogenesis on explants also did so in callus, but media containing NAA, even when effective with explants, did not cause differentiation of callus. Shoots that differentiated were excised and cultured on MS medium without growth regulators or with IBA (2.46, 4.92, 9.84 M). Roots developed after 3–8 days on an appropriate rooting medium, often without IBA. Rooted plantlets were transplanted to pots in a greenhouse and developed into normal plants. Suitable media and protocols for initiating and subculturing callus and regenerating whole plants in vitro from callus and explants have thus been established for four species of Sesbania.     

Is crassulacean acid metabolism activity in sympatric species of hemi-epiphytic stranglers such as Clusia related to carbon cycling as a photoprotective process?

A comparison of carbon isotope discrimination characteristics, crassulacean acid metabolism (CAM) activity and gas exchange together with concurrent analysis of photosystem II (PSII) chlorophyll fluorescence was conducted on leaves of sympatric species of Clusia from the restinga of Barra de Maricá, Brazil. The carbon isotope discrimination () and leaf-sap titratable acidity for leaves collected in the field indicated that the carbon metabolism of one species, C. lanceolata, was predominantly C3-like, and a second, C. fluminensis, constitutive CAM. When well-watered under glasshouse conditions C. lanceolata displayed a gas exchange pattern expected of a C3 plant, where values of instantaneous discrimination () rose from 13.5% shortly after dawn to 21.9 at midday, suggesting that all CO₂ uptake was mediated solely by ribulose 1,5-bisphosphate carboxylase (RUBISCO). C. fluminensis showed a gas exchange pattern which clearly exhibited all four phases of CAM. values during phase II ranged from –0.4 at dawn to 5.9 some 3 h later, indicating that C4 carboxylation dominated CO₂ uptake during the morning with an increasing contribution by RUBISCO, suggested by the 5 shift in at this time. The dominance of phosphoenol-pyruvate carboxylase (PEPc) activity was also found during phase IV, and extended throughout the dark period (phase I) in C. fluminensis, such that values of measured were negative (–5.0 to –0.4). This is the first time that negative values have been reported, close to those predicted theoretically for PEPc activity. The day-time uptake of CO₂ mediated by PEPc could lead to futile cycling through RUBISCO. In C. fluminensis organic acids were subjected to carbon turnover between PEPc and RUBISCO during phase II of CAM, serving perhaps to dissipate ATP and reductant at a time when excess photons are absorbed. Under low levels of photosynthetically active radiation (PAR) the two species displayed similar chlorophyll fluorescence characteristics, although for the CAM C. fluminensis a lower rate of decarboxylation of acids in the afternoon was reflected in changed quenching capacity. Under high PAR both species responded directly to changes in incident radiation, reflected by decreases in photon use efficiency (PSII) and the intrinsic photochemical efficiency (F _V/F _M), together with high and reversible quenching of excess light by the means of radiationless or thermal dissipation (q _N). Both species, with such markedly different carboxylation characteristics achieve similar rates of electron transport and maintain photosynthetic integrity. Under field conditions, however the severity of a prolonged dry season caused the CAM species to become deciduous, whereas the C3-like species remained healthy. This suggests that the widely expected advantages of CAM do not extend to tolerance of extreme environmental conditions, in contrast to the more C3-like of these sympatric species.     

Predation by a water mite (Piona exigua) on enclosed populations of zooplankton

Short term, replicated experimental alteration of densities of a predatory water mite Piona exigua Viets, inside 0.45–0.475 m³ litre enclosures, revealed little evidence of the effects of predation on the number and relative abundance of the enclosed zooplankton species. Predation rates more closely approximated those estimated from single prey functional response experiments in the second experimental period (December) than in the first (March). In December Daphnia was the only susceptible taxon present in large numbers, whereas in March, Ceriodaphnia and Chydorus were also present. This result is consistent with laboratory findings that predation rates are lowered in the presence of more than one prey type.The difficulty of obtaining evidence for significant effects of these planktonic predators is in part due to changes in the preferred prey species in the diet of Piona depending on stage and sex of the mite and to aspects of experimental design. The wide variability between replicate enclosures at each predator density reduced the power of the statistical analyses used to test the null hypothesis. Enclosures with no predators are necessary to investigate the effects of enclosure on the zooplankton prey, since these effects may outweigh those due to predator consumption.     

What makes a species vulnerable to extinction? Comparative life-history traits of two sympatric snakes

Although it is well known that species vary in their vulnerability to extinction, the reasons are poorly understood. Theory predicts that long-lived species with slow life histories (small litters, slow growth, late maturation) should be at greater risk than short-lived species with high potential rates of increase. This hypothesis was tested by comparing life-history traits of two species of sympatric, elapid snakes: the endangered broad-headed snake, Hoplocephalus bungaroides, and common small-eyed snake, Cryptophis nigrescens. From 1992 to 2000 a mark–recapture study of both species was undertaken in Morton National Park, south-eastern Australia, and this information was used to construct transition matrices for each species. The endangered H. bungaroides was found to mature late (6 years of age), had a high juvenile (54.7%) and adult (81.6%) survival rate, and a long generation length (10.4 years). In striking contrast, the common C. nigrescens matured early (within 3 years), had a lower juvenile (30.4%) and adult (74.4%) survival rate (but higher recruitment rate), and a substantially shorter generation length (5.9 years). Elasticity analyses revealed that H. bungaroides was considerably more sensitive to survival past the age of 2 years (68.6%) than C. nigrescens (37.4%). These results provide support for the hypothesis that species with slow life histories are more vulnerable to extinction.     

Theβ-subunit of human chorionic gonadotropin containsN-glycosidic trisialo tri- and tri′-antennary carbohydrate chains

TheN-linked carbohydrate chains of the-subunit of highly purified urinary human chorionic gonadotropin have been re-investigated. The oligosaccharides were released enzymatically by peptide-N ⁴-(N-acetyl--glucosaminyl)asparagine amidase-F, and fractionated by a combination of FPLC and HPLC. As a result of the application of improved fractionation methods, apart from the earlier reported carbohydrate chains, also small amounts of trisialo tri- and tri-antennary oligosaccharides were found. The primary structures of the latter carbohydrate chains have been determined by 500-MHz¹H-NMR spectroscopy to beAbbreviations hCG human chorionic gonadotropin - hCG- -subunit - hCG- -subunit - PNGase-F peptide-N ⁴-(N-acetyl--glucosaminyl)asparagine amidase-F (E.C. 3.5.1.52) - endo-F endo--N-acetylglucosaminidase-F (E.C. 3.2.1.96) - SDS sodium dodecyl sulphate - PAGE polyacrylamide gel electrophoresis - CBB coomassie brilliant blue R 250 - GlcNAc N-acetylglucosamine - NeuAc N-acetylneuraminic acid - Man mannose - Gal galactose - Fuc fucose