Genetic diversity among populations and size classes of buckeyes (Aesculus: Hippocastanaceae) examined with multilocus VNTR probes

 Little is known about genetic variation in members of the genus Aesculus (Hippocastanaceae), in particular A. flava (yellow buckeye) and A. glabra (Ohio buckeye). Here, three synthetic DNA probes (composed of tandemly repeated, core sequences) that reveal alleles at multiple variable-number tandem-repeat (VNTR) loci in these two species were used to investigate: 1) levels of genetic variation in one stand of A. flava and three isolated stands of A. glabra; 2) whether the stands of A. glabra are genetically differentiated from one another; 3) whether there has been selection for more heterozygous individuals through time in one stand each of A. flava and A. glabra; and 4) whether a possible genetic bottleneck had occurred during the formation of either species of Aesculus. First, variation of VNTR genetic markers within and among three populations of A. glabra separated by 60–180 km was examined. In each one hectare (ha) population, 22 individuals were randomly sampled. Among the three populations, the mean number of bands scored per individual was 80.35 and the average number of estimated loci surveyed was 54.17. Mean similarity and estimated heterozygosity within populations ranged from 0.634 to 0.743 and from 0.342 to 0.486, respectively. The mean similarity across populations was 0.657, while the mean estimated heterozygosity across populations was 0.484 for A. glabra. The most isolated site was the most genetically differentiated as indicated by differences in levels of similarity, heterozygosity, and Fst value comparisons. In a separate experiment, genetic variation in 22 large (reproductively mature; dbh > 8 cm) individuals was compared with that in 22 small (not yet reproductive; dbh < 1 cm) individuals collected within one ha stands for both A. flava and A. glabra. Mean similarity values among large versus small individuals of A. flava were 0.665 versus 0.662, while for A. glabra the corresponding values were 0.686 versus 0.691, respectively. Permutation tests of these similarity data detected no evidence for size class genetic differentiation in either species (both p-values > 0.050). Further, permutation tests for the number of bands per individual (average band number should be higher in more heterozygous individuals) detected no significant differences between size classes for either species. Thus, evidence of pronounced inbreeding and/or selection altering population genetics within small relative to large individuals was not detected. In addition, comparable similarity and heterozygosity values between these two closely related species (which still maintain an active zone of hybridization) suggests that either: 1) no extreme genetic bottleneck has accompanied the formation of these species from a common ancestor; or 2) signs of such a bottleneck have largely been eliminated. These studies demonstrate the utility of multilocus VNTR DNA probes for investigating genetic variation within and among plant populations, between size classes within a population, and between closely related species. Received May 15, 1998 Accepted September 11, 2001     

Relationships between nuclear DNA content and seed and leaf size in soybean

 A correlation between genome size and agronomically important traits has been observed in many plant species. The goal of the present research was to determine the relationship between genome size, seed size, and leaf width and length in soybean [Glycine max (L.) Merr.] Twelve soybean strains, representing three distinct seed size groups, were analyzed. Flow cytometry was used to estimate their 2C nuclear DNA contents. Data on seed size and leaf size of the 12 strains were obtained from 1994 and 1995 field experiments. Variation of 2C nuclear DNA among the 12 soybean strains was 4.6%, ranging from 2.37 pg for a small-seed strain to 2.48 pg for a large-seed strain. Strain seed size was positively associated with leaf width (r=0.92) and leaf length (r=0.93). Genome size was highly correlated with seed size (r=0.97), leaf width (r=0.90) , and leaf length (r=0.93). The results of our study indicate that there is a significant correlation between genome size and leaf and seed size in soybean. It is possible that selection for greater seed size either leads to, or results from, greater genome size. If so, this relationship might be worth exploring at a more fundamental level. Received: 5 April 1997 / Accepted: 9 January 1998     

Spawning activities and physical characteristics of the spawning ground of Lethenteron reissneri at the headstream of the Himekawa River, central Japan

 The spawning period of the Far Eastern brook lamprey, Lethenteron reissneri, in the headstream of the Himekawa River is estimated to be between mid-March and late May with the peak of spawning activity between early April and early May. The sex ratio (female:male) in 1999 ranged from 1 : 2.5 to 1 : 3.0 (mean 1 : 2.8) and in 2000 from 1 : 0.8 to 1 : 4.0 (mean 1 : 2.4). In >90% of the observations of spawning nests, males outnumbered females. The construction area of spawning nests tended to shift upstream during the spawning period. The nests were constructed at water depths between 5 and 70 cm, water velocity between 10 and 30 cm/s, and on substrate with pebbles of 5–20 mm in diameter. Lethenteron reissneri constructed nests on substrate similar with Petromyzon marinus, but at shallower points and in areas with a slower water velocity. Received: April 2, 2001 / Revised: December 12, 2001 / Accepted: December 27, 2001     

Effect of masking the parietal eye on the diurnal activity and body temperature of two sympatric species of monitor lizards, Varanus s. salvator and Varanus b. nebulosus

 Five water monitor lizards, Varanus salvator salvator, and four clouded monitor lizards, Varanus bengalensis nebulosus, were caught on Tioman island in Malaysia. A radio-thermistor transmitter was implanted into the buccal cavity of each animal, and they were released into an enclosure measuring 5.5 × 6.5 metres. The lizards were observed for 9 and 8 days, respectively, before and after the parietal eye was covered with aluminium foil. With uncovered parietal eye, both species showed a clear diurnal rhythm, being active only during day time. After covering the parietal eye, the mean locomotor activity of five V. s. salvator decreased from 791 to 107 min · day^–1 but remained unchanged around 850 min · day^–1 for V. b. nebulosus. The mean duration of locomotor activity decreased in V. s. salvator and V. b. nebulosus after the parietal eye was covered, but V. b. nebulosus maintained its locomotor activity by increasing the number of locomotor bouts. The water monitor spent very little time on thermoregulation. Its body temperature ranged between 26.3 and 28.4 °C, which decreased after the parietal eye was covered. The clouded monitor thermoregulated around 28.8–36.0 °C, which remained unchanged after the parietal eye was covered. In both species, there was a strong correlation between body temperature and ambient temperature. Behavioural abnormalities were recorded among V. s. salvator with covered parietal eye. They were often observed to be active by night and often slept outside a burrow. The circadian rhythm of V. b. nebulosus appeared unaffected by shielding of its parietal eye. Captivity combined with shielded parietal eye induced agonistic behaviour in both species. Accepted: 11 September 1996     

Morphological differences among three species of newly settled pocilloporid coral recruits

 Investigation of the life history of corals is hampered by an inability to identify early recruits. In this study, the pattern of formation and morphology of the juvenile skeletons of three laboratory-reared pocilloporids, Seriatopora hystrix, Stylophora pistillata and Pocillopora damicornis, were compared to determine whether they could be reliably distinguished. The pattern of skeleton formation, including the origin and structure of the septa, columella and corallite wall was similar in all species. Following the completion of the primary corallite wall after 4–5 days, these species could be identified by differences in the diameter of the primary corallite. The mean diameter (±SE) of each species differed markedly: S. hystrix 400 ± 2.7 μm, range 325–450 μm; S. pistillata 505 ± 3.5 μm, range 400–550 μm; P. damicornis 697 ± 7.5 μm, range 492–885 μm. Values for the primary corallite diameter overlapped in only 3% of samples, demonstrating the potential utility of this feature as a tool for classifying recruits obtained from the field. Accepted: 4 January 2000     

Upeneus australiae, a new goatfish (Mullidae: Perciformes) from Australia

 Upeneus australiae sp. nov. is described on the basis of 14 specimens collected from Australian waters. Although it belongs to the seven-spined group of Upeneus, now comprising five species, U. australiae is distinguished from its congeners by the combination of the following characters: 6–7 + 16–18 (modally 7 + 17) gill rakers, ectopterygoids without teeth, five and six black oblique bars on upper and lower caudal lobes, respectively, and peritoneum transparent with small black spots. Received: July 26, 2001 / Revised: November 2, 2001 / Accepted: December 4, 2001     

Deletions in the plastid trnT–trnL intergenic spacer define clades within Cactaceae subfamily Cactoideae

 The phylogenetic distribution of two deletions, of about 350 and 250 bp respectively, within the chloroplast trnT–trnL intergenic spacer was examined. One deletion was found in all members of Cactaceae subfamily Cactoideae sampled, totaling 37 species, but not in taxa from other subfamilies or closely related families. The second was shared by a subset of Cactoideae comprising members of tribes Cereeae, Trichocereeae, and Browningieae (in part), as well as Harrisia. Close links among the former three South American tribes have been previously hypothesized. This distribution suggested that Browningieae, a tribe defined largely by shared primitive features, were not monophyletic, and that Harrisia may have been incorrectly placed outside the Browningieae–Cereeae–Trichocereeae group. Received June 12, 2001 Accepted October 26, 2001     

Effect of the removal of cleaner fish on the abundance and species composition of reef fish

The ecological significance of cleaner fish on coral reefs was investigated. I removed all cleaner fish, Labroides dimidiatus, from eight small reefs, measured the subsequent effect on the abundance and species composition of all reef fish after 3 and 6 months, and compared it with eight control reefs with cleaner fish. The removal of cleaner fish had no detectable effect on the total abundance of fish on reefs and the total number of fish species at both times. Multivariate analysis by non-metric multidimensional scaling and ANOSIM pairwise tests based on 191 fish species revealed no effect of cleaners on the community structure of fish. Similar results were obtained using principal components analysis on subsets of the data using the 33 most common fish species and the 15 most abundant species (≥5 individuals per reef ) with both log10 (x + 1) transformed data and with fish numbers standardized for abundance. This study demonstrates that the removal of cleaner fish for 6 months did not result in fish suffering increased mortality nor in fish leaving reefs to seek cleaning elsewhere. Received: 28 October 1996 / Accepted: 7 February 1997     

The relative importance of tryptophan-dependent and tryptophan-independent biosynthesis of indole-3-acetic acid in tobacco during vegetative growth

A quantitative study of indole-3-acetic acid (IAA) turnover, and the contribution of tryptophan-dependent and tryptophan-independent IAA-biosynthesis pathways, was carried out using protoplast preparations and shoot apices obtained from wild-type and transgenic, IAA-overproducing tobacco (Nicotiana tabacum L.) plants, during a phase of growth when the level of endogenous IAA was stable. Based on the rate of disappearance of [¹³C₆]IAA, the half-life of the IAA pool was calculated to be 1.1 h in wild-type protoplasts and 0.8 h in protoplasts from the IAA-overproducing line, corresponding to metabolic rates of 59 and 160 pg IAA (μg Chl)⁻¹ h⁻¹, respectively. The rate of conversion of tryptophan to IAA was 15 pg IAA (μg Chl)⁻¹ h⁻¹ in wild-type protoplasts and 101 pg IAA (μg Chl)⁻¹ h⁻¹ in protoplasts from IAA-overproducing plants. In both instances, IAA was metabolised more rapidly than it was synthesised from tryptophan. As the endogenous IAA pools were in a steady state, these findings indicate that IAA biosynthesis via the tryptophan-independent pathway was 44 pg IAA (μg Chl)⁻¹ h⁻¹ and 59 pg IAA (μg Chl)⁻¹ h⁻¹, respectively, in the wild-type and transformed protoplast preparations. In a parallel study with apical shoot tissue, the presumed site of IAA biosynthesis, the rate of tryptophan-dependent IAA biosynthesis exceeded the rate of metabolism of [¹³C₆]IAA despite the steady state of the endogenous IAA pool. The most likely explanation for this anomaly is that, unlike the protoplast system, injection of substrates into the apical tissues did not result in uniform distribution of label, and that at least some of the [²H₅]tryptophan was metabolised in compartments not normally active in IAA biosynthesis. This demonstrates the importance of using experimental systems where labelling of the precursor pool can be strictly controlled. Received: 18 January 2000 / Accepted 24 February 2000     

<Emphasis Type="Italic">Careproctus parvidiscus</Emphasis>, a new species of liparid fish (Teleostei: Scorpaeniformes) collected from the southern Okhotsk Sea,Japan

 A new liparid, Careproctus parvidiscus, is described on the basis of a single specimen (177 mm in standard length) collected from the southern Okhotsk Sea, off Shiretoko Point, Hokkaido, Japan, at 400–700 m depth. It is distinguished from other congeners by the following combination of the characters: 50 dorsal fin rays, 44 anal fin rays, 10 + 47 = 57 vertebrae, 2 pleural ribs, 14 pyloric caeca being slender and pointed, 2 suprabranchial pores, narrower gill opening, longer lower lobe of pectoral fin, base of uppermost pectoral fin ray almost on a level with center of eye, rudimentary disk, dusky peritoneum, and black stomach. Received: June 13, 2001 / Revised: December 7, 2001 / Accepted: December 22, 2001     

Taxonomic implications of genome size for all species of the genus Gasteria Duval (Aloaceae)

Nuclear DNA content (2C) is used as a new criterion to investigate all species of the genus Gasteria Duval including the three recently described species Gasteria polita van Jaarsv., G. pendulifolia van Jaarsv. and G. glauca van Jaarsv.. The 122 accessions investigated have the same chromosome number (2n=2x=14), with exception of three tetraploid plants found. The nuclear DNA content of the diploids, as measured by flow cytometry with Propidium Iodide, is demonstrated to range from 32.8–43.2 pg. This implies that the largest genome contains roughly 10¹⁰ more base pairs than the smallest. Based on DNA content the species could be divided in five groups: G. rawlinsonii Oberm. with 32.8 pg, 13 mostly inland species with 34.3–36.0 pg, five coastal species with 36.5–39.0 pg and Gasteria batesiana Rowley with 43.2 pg. The thirteen species with 34.3–36.0 pg could be divided further, in a group of eight species occupying mainly very restricted areas with 34.3–35.1 pg and a second group of five species with 35.2–36.0 pg mainly occupying large areas. These five groups did not coincide very well with the two sections and four series of Gasteria based on a cladistic analysis by van Jaarsveld et al. (1994). Based on its long leafy branches, location in the centre of Gasteria species distribution and its by far lowest DNA content, G. rawlinsonii might be the most primitive member of the genus. Nuclear DNA content as measured by flow cytometry is shown to be relevant to provide additional information on the relationships between Gasteria species.     

Habitat association and depth distribution of two sympatric groupers of the genus Cephalopholis (Serranidae: Epinephelinae)

 Habitat association and depth distribution of two sympatric coral reef groupers of the genus Cephalopholis were examined at Rota, Mariana Islands. The two species are similar in body size, morphology, and social organization. In this study, they differed in their association with habitat and microhabitat and in depth distribution. Cephalopholis spiloparaea occurred on the reef slope between the reef terrace and deep sand flats at depths between 15 and 26 m. This species was associated mainly with Porites rus corals. Cephalopholis urodeta occurred largely on the upper reef terrace at 1–12 m. This species was associated mainly with coral pavement. The observed pattern of segregation might be the result of competitive or noncompetitive interactions or of phylogenetic constraints, but the exact mechanism or combination thereof remains unknown. Received: May 30, 2000 / Revised: September 5, 2001 / Accepted: October 25, 2001     

Differences among plant species in cuticular permeabilities and solute mobilities are not caused by differential size selectivities

Solute mobilities in cuticular membranes of six species (Hedera helix, Malus domestica, Populus alba, Pyrus communis, Stephanotis floribunda, Strophantus gratus) were measured using plant hormones, growth regulators and other organic model compounds varying in molar volumes from 99 to 349 mL · mol⁻¹ The dependence of mobilities (k*) on molar volume (V _x ) was exponential and could be described with equations of the type log k*=log k*⁰−^′ V _x . The y-intercepts (log k*⁰) represent mobilities of a hypothetical solute of zero molar volume. The parameter β′ is a measure of size selectivity of cuticular membranes and no differences among the six species were observed. At 25 °C the average β′ was 0.0095 mol · mL⁻¹. Solute mobility decreased by about a factor of 8.9 when molar volume increased by 100 mL · mol⁻¹ and the mobility of a compound with V _x  = 100 mL · mol⁻¹ was about 700-fold higher than the mobility of a compound with V _x  = 400 mL · mol⁻¹. Size selectivity decreased with increasing temperatures and for Strophantusβ′-values of 1.6 × 10⁻² to 8.0 × 10^-4 mol · mL⁻¹ were obtained for 10 and 30 °C, respectively. The-intercepts (log k*⁰) differed among plant species by 3 orders of magnitude and since size selectivity was the same for all species, solute mobilities for solutes having zero molar volumes were the sole cause for differences among species in solute mobilities and permeabilities. We argue that these differences in k*⁰ are related to tortuosity of the diffusion path. These results were used to derive an equation which predicts rates of cuticular penetration on the basis of k*⁰, the average size selectivity of 9.5 × 10⁻³ mol · mL⁻¹ and the driving forces of penetration. Received: 25 November 1997 / Accepted: 9 March 1998     

Relationship between parental chromosomic contribution and nuclear DNA content in the coffee interspecific hybrid C. pseudozanguebariae×C. liberica var ‘dewevrei’

 F₁ hybrids were obtained between two coffee species with the same chromosome number (2n=22) but with different nuclear DNA contents [C. pseudozanguebariae (PSE) 2C=1.13 pg and C. liberica var ‘dewevrei’ (DEW) 2C=1.42 pg]. G2 hybrids were obtained by open-pollination of the F₁ hybrids. Genomic in situ hybridisation (GISH) and flow cytometry were used on six F₁ hybrids and seven G2 hybrids to determine their parental chromosomic contribution and their nuclear DNA content (qDNA), respectively. GISH efficiently identified chromosomes from both species. F₁ hybrids had a qDNA intermediate between that of the parental species and contained the expected 11 chromosomes from each species. There was a linear relationship between the number of PSE chromosomes and the nuclear DNA content, which indicates that flow cytometry can be used to give a rough estimate of the parental chromosomic contribution in G2 hybrids. Received: 1 August 1997/Accepted: 25 August 1997     

Stability of diatom composition in a variable lake environment: Lake Chascomús, Argentina

 Water and surficial sediment samples of Lake Chascomús and its tributaries were analyzed in order to relate changes in diatom community structure to chemical variables. Over the course of 13 months of sampling, the lake exhibited major changes in water level (1.15–1.98 m average depth), total dissolved solids (821–1972 mg l⁻¹), silica (0.098–8.22 mg l⁻¹), and total algal biomass (21.4–145.9 μg Chl a l⁻¹). However, despite these large fluctuations, the diatom species composition was relatively stable. The dominant species in the water column was always Synedra berolinensis (68.9%–90.1% total frustules), with Fragilaria construens and F. brevistriata as subdominants. In the sediments the latter two species dominated the frustule counts. These results indicate an unusual floristic stability of this eutrophic ecosystem, with persistent dominance by broadly tolerant, generalist species. Received: April 24, 2001 / Accepted: January 23, 2002     

Cytogenetics of new Guizotia Cass. (Compositae) interspecific hybrids pertaining to genomic and phylogenetic affinities

 The following crosses were made using four recognized species/subspecies and a new population of Guizotia, referred to as Chelelu after the name of the locality in Ethiopia from which it was collected: G. scabra subsp. schimperi × Chelelu, Chelelu × G. scabra subsp. scabra, G. zavattarii × G. arborescens and Chelelu × G. zavattarii (all accessions with 2n=30). Plant morphology as well as mitotic and meiotic chromosome analysis confirmed the hybrid nature of the obtained progeny. At metaphase I of meiosis, the F₁ hybrid plants (2n=30) showed a mean of about 95%, 31%, 63% and 0.50% of the pollen mother cells with 15 bivalents, and a mean of about 14.95, 13.75, 14.40 and 7.86 bivalents per cell, respectively. The respective mean pollen stainability was about 67%, 19%, 31% and 2%. From the results it was concluded that Chelelu is more closely related to G. scabra subsp. schimperi than to G. scabra subsp. scabra but more to the latter than to G. zavattarii. Guizotia zavattarii and G. arborescens are closely related to each other. Based on the cytological observations made, the probable basic chromosome number for the genus is discussed. Received November 5, 1999 Accepted August 21, 2001     

Effects of above-ground browsing by mammals on mycorrhizal infection in an early successional taiga ecosystem

Using an exclosure experiment in the willow stage of primary succession on the floodplain of the Tanana River, we tested the hypothesis that browsing can reduce mycorrhizal infection. We measured the effects winter browsing by moose (Alcesalces) and snowshoe hare (Lepusamericanus) had on mycorrhizal infection and fine root biomass of willow (Salix spp.) and balsam poplar (Populusbalsamifera). We found that protection from winter browsing increased ectomycorrhizal infection by 10% in the top 5 cm of the soil profile, by 23% at 5–10 cm, and by 42% at the 10–15 cm depth. Mammal browsing in taiga forests is now recognized as a major cause of the shift from palatable deciduous species such as willow and balsam poplar to less palatable species such as alder and spruce. We suggest that browsing-induced reduction in ectomycorrhizal infection of salicaceous species plays a central role in this shift in plant community composition. Received: 26 March 1996 / Accepted: 26 September 1996     

Chromosomal differentiation and genome size in three European mountain Lilium species

 Three related and taxonomically close species of the genus Lilium (L. pyrenaicum Gouan, L. pomponium L. and L. carniolicum Bernh.), all of them with 2n=24 chromosomes, have been studied for chromosomal differentiation, using fluorochrome banding and fluorescence in situhybridization (FISH), and for genome size and GC percentage using flow cytometry. The total DNA content of L. pomponium (2C=70.26 pg) was about 5% higher than that of L. pyrenaicum (2C=67.74) and L. carniolicum (2C=67.37 pg), while GC percentage was higher in this last species (36.60%) than in L. pomponium (35.56%) and lower than in L. pyrenaicum (37.92%). Silver staining, fluorochrome banding with chromomycin A₃ (CMA) and fluorescence in situ hybridization (FISH) clearly pointed out the number of nucleoli, the number and position of GC-rich bands and the number and location of rDNA sites thus permitting distinction of the three species at chromosomal level. Two families of ribosomal genes, 18S-5.8S-26S (18S) and 5S rRNA genes, were separated onto different pairs in chromosome complements of examined species. Chromosome regions containing both kinds of rRNA genes were also GC-rich regions. The results revealed a clear interspecific differentiation at the chromosomal level and permitted the discussion about relationships among the species. Received June 21, 2002; accepted October 4, 2002 Published online: Febraury 7, 2003     

Early ontogeny of Lophonectes gallus (Bothidae) from southeastern Australia

 The early ontogeny of Lophonectes gallus (Bothidae) is described based on 83 specimens (1.9–17.5 mm BL), collected from the Tasman Sea off southeastern Australia. The larvae are diagnosed by the following array of characters: vertebrae 10 + 30–31 = 40–41; one elongated dorsal fin ray and several melanophores present on gut in preflexion stage (1.9–4.7 mm BL); and spines on posterior basipterygial process, and urohyal, cleithrum, and epiotic without spines after postflexion stage (8.0–17.5 mm BL). The larvae are relatively small at metamorphosis (15–18 mm BL) compared with other bothid larvae. Received: March 22, 2001 / Revised: December 12, 2001 / Accepted: December 26, 2001     

Genome size and cytogenetic characterization of three Algerian Retama species

Thirty-three populations belonging to the three Retama species, Retama monosperma, Retama raetam and Retama sphaerocarpa, were collected to study species differentiation using flow cytometry for 2C DNA assessment and molecular cytogenetics for karyotype organisation. All were 2n = 48. Genome size ranged from 1.76 to 1.97 pg and revealed significant intraspecific variation correlated to the geographic distribution of the populations. The number and position of the two ribosomal gene families 5S and 45S were determined by fluorescent in situ hybridization, revealing chromosome reorganisation between species. In R. raetam and R. monosperma, the minor 5S loci co-localised with 45S on the satellite chromosome pair. Fluorochrome banding identified GC- and AT-rich DNA regions. In R. monosperma a unique chromomycin positive GC-rich band was observed associated with the secondary constriction. In contrast, an original pattern showing two chromomycin positive bands localised at each side of the extended rDNA locus was observed in R. sphaerocarpa and R. raetam. The polymorphism revealed in our cytogenetic data allowed us to separate the group of R. raetam and R. monosperma from R. sphaerocarpa.