Interspecific Social Interactions and Breeding Success of the Frog Rana latastei: A Field Study

Interspecific reproductive interference can affect fitness‐related breeding performance, thus influencing fitness and distribution of populations. Laboratory studies demonstrated the social interference of Rana dalmatina males on R. latastei breeding females: the presence of heterospecific males reduced the percentage of viable embryos in R. latastei eggs. Here, we tested if the negative effects of R. dalmatina males on R. latastei reproductive success occur in field conditions. We compared the percentage of viable embryos of eggs laid in field conditions from populations where R. latastei breeds alone with the percentage of viable embryos of populations where R. latastei cohabits with R. dalmatina. We did not find any significant difference in percentage of viable embryos between R. latastei populations syntopic and allotopic with R. dalmatina, nor a relationship between the relative abundance of heterospecifics and reproductive success. In natural conditions, the presence of heterospecific males does not seem to interfere with the reproductive success of R. latastei. The experimental procedure may influence the interaction among individuals. Therefore, we suggest to validate on natural populations the results of experiments dealing with complex interactions.     

Physical mapping of ribosomal DNA on several species of the subgenus Rosa

The localization of NORs by fluorescent in situ hybridization on metaphase spreads of five diploid (Rosa gigantea Coll., Rosa moschata Herrm., Rosa multiflora Thunb., Rosa rugosa Thunb. and Rosa sempervirens L., 2n=2x=14), one triploid (Rosa chinensis’semperflorens’ Koehne., 2n=3x=21) and one tetraploid (Rosa gallica ’versicolor’ L., 2n=4x=28) ancestral species of modern roses was studied. Two terminal hybridization signals were observed in all diploid species corresponding to a single NOR per genome in these species. Triploid R. chinensis showed three hybridization sites on the short arm of three morphologically similar chromosomes. Six hybridization sites, located at terminal positions of the short arms of three chromosome pairs, were observed in the tetraploid species. These signals corresponded to three pairs of NORs and all of them were located in chromosome pairs of different size. These observations, together with the analysis of meiotic pairing in PMCs, support the view that our specimen of R. chinensis is an autotriploid and that R. gallica’versicolor’ has an alloploidy nature. Received: 27 November 2000 / Accepted: 12 March 2001     

Speciation in the Rana chensinensis species complex and its relationship to the uplift of the Qinghai-Tibetan Plateau

Speciation remains a fundamental issue in biology. Herein, we report an investigation into speciation in the Rana chensinensis species complex using DNA sequence data from one mitochondrial and five nuclear genes. A phylogenetic analysis of the data revealed four major clades in the complex, and each of them was found to likely represent a species, including one cryptic species. Ecological niche models were generated from 19 climatic variables for three of the four major clades, which were represented by widespread sampling, including R. chensinensis, Rana kukunoris and the potential cryptic species. Each clade is associated with a unique ecological unit, and this indicates that ecological divergence probably drove speciation. Ecological divergence is likely related to the late Cenozoic orogenesis of the Qinghai–Tibetan Plateau. In addition, gene flow between species was detected but only in peripheral portions of the ranges of the four major clades, thus likely had little influence on the speciation processes. Discordances between mitochondrial and nuclear genes were also found; the nominal species, R. chensinensis, contains multiple maternal clades, suggesting potential mitochondrial introgression between R. chensinensis and R. kukunoris.     

Banding patterns of the chromosomes ofCercopithecus neglectus: Comparison withMiopithecus talapoin andErythrocebus patas

The G, Q, and C bands and the location of the nucleolar organizing regions (NORs) of the chromosomes of two male Cercopithecus neglectusare described. The diploid number of the species is 2n =62. Comparison with the karyotypes of Miopithecus talapoin (2n =54), and Erythrocebus patas (2n =54)showed the presence of total banding homeology for only 10 chromosome pairs.     

Conservation of chromosomal location of nucleolus organizer in American marsupials (Didelphidae)

The distribution and expression of nucleolus organizer regions (NORs) were analyzed in seven species of marsupials representative of the three karyotypes (2n = 14, 18 and 22) found in the American family Didelphidae. Analyses comprised silver-staining of NORs and fluorescence in situ hybridization with an rDNA probe. In addition to confirming the variability in number and distribution of NORs in Didelphidae, we demonstrated the conserved location of NORs on one autosome pair in the three karyotypes. In Monodelphis domestica (2n = 18), the NOR on the X chromosome was not inactivated in females.     

Social environment and reproductive interference affect reproductive success in the frog Rana latastei

Variation in the social environment produces selection on morphologicaland behavioral traits. It is less clear how the social environmentgenerates variation in demography through behavioral mechanisms.Theory suggests that one aspect of social environment, relativeabundance of heterospecifics, influences the intensity of reproductiveinterference and its demographic effects. These effects arecountered by species recognition and female preferences. Westudied the effects of social environment on reproductive successin replicated, mixed breeding populations of two ranid frogs,Rana latastei and Rana dalmatina, the former being of internationalconservation concern. We manipulated the social environmentof female R. latastei experimentally by varying the relativeabundance of potential conspecific and heterospecific sexualpartners. We measured amplexus frequency and recorded the reproductivesuccess of R. latastei females. When conspecific males wererelatively uncommon, (1) the absolute and relative frequenciesof conspecific amplexus decreased, indicating a breakdown ofsexual isolation, (2) oviposition was less frequent, and (3)the percentage of viable embryos in deposited clutches decreased.R. latastei females in an environment of low relative conspecificabundance (1:5, R. Latastei:R. dalmatina) demonstrated 6.8%the reproductive success of females in an environment exclusivelywith conspecifics. We present a model for the dependence ofconspecific amplexus on the social environment. We discuss severalmechanisms that may influence reproduction by R. latastei, andwe support conservation of the species' preferred habitat toreduce opportunities for reproductive interference that occurat shared breeding sites.     

Susceptibility of Italian agile frog populations to an emerging strain of Ranavirus parallels population genetic diversity

Western populations of the Italian agile frog (Rana latastei) experience widespread genetic depletion. Based on population genetic theory, molecular models of immunity and previous empirical studies, population genetic depletion predicts increased susceptibility of populations to emergent pathogens. We experimentally compared susceptibility of R. latastei populations upon exposure to an emerging strain of Ranavirus, frog virus 3 (FV3), using six populations spanning the geographical range and range of population genetic diversity found in nature. Our findings confirm this prediction, suggesting that the loss of genetic diversity accompanying range expansion and population isolation is coincident with increased mortality risk from an emergent pathogen. Loss of heterozygosity and escape from selection imposed by immunologically cross‐reactive pathogens may potentially generate range‐wide variation in disease resistance.     

Comparative analysis of rDNA location in five Neotropical gomphocerine grasshopper species

We report here, for the first time, the chromosome complement, number and location of the nucleolar organizer regions (NORs) revealed by silver staining (AgNO₃) and fluorescent in situ hybridization (FISH) in five Neotropical gomphocerine species: Rhammatocerus brasiliensis, R. brunneri, R. palustris, R. pictus and Amblytropidia sp. The objective of this study was to summarize available data and propose a model of chromosome evolution in Neotropical gomphocerines. All five species studied showed chromosome numbers consisting of 2n = 23,X0 in males and 2n = 24,XX in females. Amblytropidia sp. was the only species showing a bivalent (M₈) with megameric behavior during meiosis. The rDNA sites were restricted to autosomal pairs, i.e. the pericentromeric region of the S₉ chromosome, the consensus NOR location in all five species. R. brasiliensis was the only species showing additional NORs on M₄ and M₆ pairs which, likewise the S₉ NOR, were active in all cells analyzed. Comparison of these results with those reported previously in Palearctic gomphocerine species suggests higher resemblance of Neotropical species with the Old World species also possessing 23/24 chromosomes. Evolutionary mechanisms responsible for the observed interspecific variation in NOR location in this group are discussed.     

Karyotype analysis,nucleolus organizer regions and C-banding pattern of Eisenia foetida (oligochaeta,lumbricidae)

The diploid number 2n=22 and haploid number n=11 found for Eisenia foetida from Palermo, Italy, confirm earlier data for this species from other localities. Analyses of silver-stained and C-banded mitotic and meiotic chromosomes suggest that a single chromosome pair has active NORs which correspond with C-positive regions. The occurrence of nucleolus activity during spermatogenesis of E. foetida is ascertained.     

Supernumerary chromosomes in a Rhamdia hilarii population (Pisces,Pimelodidae)

A study was conducted on the chromosomes of a Rhamdia hilarii (Pisces, Pimelodidae) population. The results suggest that the basic chromosome number is 2n=58, with numerical variation up to a limit of 2n=63, due to the presence of supernumerary chromosomes which seem to be mitotically stable. These chromosomes are metacentrics and can be different in size. The C-banding pattern, showing heterochromatin especially in both telomeric regions, permits their identification in the karyotype. The NORs are located on secondary terminal constrictions on the short arm of a pair of subtelocentric chromosomes. However, there may be heteromorphism in the size of the secondary constrictions and, consequently, in the size of the NORs.     

Karyological studies in four species of ardeid birds (Ardeldae,Ciconiiformes)

Karyotypic analyses of 4 species of birds: Ardeola grayii grayii, Bubulcus ibis coromandus, Egretta garzetta, Nycticorax nycticorax nycticorax (Ardeidae, Ciconiiformes) reveal low diploid values (2n=60–68), absence of sharp bimodality and an unusually large W chromosome in A. g. grayii. N-banding studies in two species (B. i. coromandus and N. n. nycticorax) show 6–8 microchromosomes to contain NORs and the G-banded karyotype of A. g. grayil reveals the usual avian pattern.     

Distribution of rDNA loci in the genus Glycine Willd.

The objective of this study was to examine the distribution of rDNA loci in the genus Glycine Willd. by fluorescent in situ hybridization (FISH) using the internal transcribed spacer (ITS) region of nuclear ribosomal DNA as a probe. The hybridized rDNA probe produced two distinct yellow signals on reddish chromosomes representing two NORs in 16 diploid (2n=40) species. Aneudiploid (2n=38) and aneutetraploid (2n=78) Glycine tomentella Hayata also exhibited two rDNA sites. However, the probe hybridized with four chromosomes as evidenced by four signals in two diploid species (Glycine curvata Tind. and Glycine cyrtoloba Tind.) and tetraploid (2n=80) G. tabacina (Labill.) Benth. and G. tomentella. Synthesized amphiploids (2n=80) of Glycine canescens F. J. Herm. (2n=40) and the 40-chromosome G. tomentella also showed four signals. This study demonstrates that the distribution of the rDNA gene in the 16 Glycine species studied is highly conserved and that silence of the rDNA locus may be attributed to amphiplasty during diploidization and speciation. Received: 10 October 2000 / Accepted: 6 December 2000     

Localization of NORs in spermatogonial metaphase chromosomes of six species of grasshoppers

The silver staining technique was employed to locate Nucleolar Organiser Regions (NORs) in six species of grasshoppers viz. Aiolopus thalassinus F. (Tryxalinae); Oeodaleus abruptus Thunb., Gastrimargus transversus Thunb., Heteropternis respondens Walk. (Oedipodinae); Parahieroglyphus biliniatus Bol. and Spathosternum prasiniferum Walk. (Catantopinae). Usually the NORs were located on the larger elements of the chromosomal complement. However, in O. abruptus NORs were found on autosomes S₈ and S₉. The salient observations were: (1) NORs were seen in only a few of the several spermatogonial metaphases examined; (2) Active NORs were mostly located either on one chromatid of the homologues or on the homologue depicting heteromorphism; (3) NORs showed either proximal, subproximal or interstitial locations. However, in O. abruptus and P. bilineatus NORs were located at two positions. Distribution of NORs in different species and their probable role in tracing the evolutionary pathways in Acridoidea are discussed.     

Elimination of Solanum phureja nucleolar chromosomes in S. tuberosum + S. phureja somatic hybrids

Summary The karyotype of the dihaploid SVP1 line of S. tuberosum (2n=2x=24) showed two nucleolar chromosomes with differently sized satellites. The diploid SVP5 line (2n=2x=24) and tetraploid regenerants of S. phureja had larger but similar satellites. Somatic hybrids between the diploid lines of these potato species with genome combinations 4 tub + 2 ph (plants 1–3), 2 tub + 4 ph (plants 4–7) and 4 tub + 4 ph (plant 8) had lost 2 phureja nucleolar chromosomes if 4 phureja genomes were present. One phureja nucleolar chromosome of plants 1–3 and both of plants 5 and 7 had rearranged satellites. Elimination of the two nucleolar chromosomes occurred preferentially, was under genetic control, and probably took place during early callus development. NOR activity resulting in rear-rangements between NORs may have caused the elimination.     

Karyological investigations inRanunculus subg.Batrachium (Ranunculaceae) on the Aegean islands

This paper is the first report of an investigation on differentiation and evolutionary pattern inRanunculus subg.Batrachium mainly on the Aegean islands and treats the karyology in the subgenus. The following taxa and chromosome numbers are recorded.R. peltatus subsp.peltatus: 2n = 32, subsp.baudotii: 2n = 32, subsp.saniculifolius: 2n = 16, 32, and 40;R. peltatus s.l.: 2n = 32 and 48;R. penicillatus subsp.pseudofluitans: 2n = 16;R. rionii: 2n = 16;R. sphaerospermus: 2n = 16;R. trichophyllus: 2n = 32, 40, and 48;R. tripartitus: 2n = 48. The karyotypes show certain interspecific differences in chromosome size, in number of chromosome types according to centromere position and in occurrence and position of satellite chromosomes. r-Index and RCL have been calculated.     

C-banding patterns in the AustralianBulbine (Liliaceae): The perennial group,B. bulbosa s. lato

C-banding studies support earlier evidence thatB. bulbosa, as a previously circumscribed, is heterogeneous, consisting of three distinct entities: (1) theB. bulbosa complex (B. bulbosa s. str.) at 4x (2n = 24), 8x (2n = 48) and 12x (2n = 72) ploidy levels, (2) the rock lily and (3) the Kroombit population (both 2n = 46). Each of these three main groups has a distinctive banding profile, though centromeric and telomeric dot bands, variably expressed, are common to all. In theB. bulbosa complex, substantial heterochromatin development, apart from bands associated with the NORs on chromosomes 1 L, 2 S and 3 L, occurs only at the terminal regions of the short arms of the large and middlesized acrocentric chromosomes, with considerable polymorphic and polytypic variation in the number and size of the heterochromatic blocks, especially at the 4x level. Queensland 8xB. bulbosa populations differ in having terminal heterochromatin, probably associated with NORs, on 11 S and 12 S, and in having some strong interstitial bands. The differences appear to correlate with attributes relating to flower morphology, and may have systematic significance. The karyotypes of rock lily and Kroombit are somewhat similar but the former has a characteristic C-band profile with multiple interstitial bands on chromosomes 1–5 and 7–9, whereas the latter has only one interstitial band on chromosome 9.First contribution of a series on cytoevolution in the AustralianBulbine. Two introductory papers in Austral. J. Bot.34 (2)     

Analysis of heterochromatin by combination of C-banding and CMA3 and DAPI staining in two fish species (Pimelodidae,Siluriformes)

The chromosomes of Steindachneridion sp. (2n = 56) and Rhamdia quelen (2n = 58) were analyzed by C-banding (CB) and Chromomycin A₃ (CMA₃) and 4,6-diamidino-2-phenylindole (DAPI) staining, separately and consecutively, in order to understand the role of base-specific fluorochrome treatment after CB. Both species' chromosomes shared common staining profiles as follows. CB with Giemsa (CBG) revealed weak heterochromatic blocks in the telomeric regions of some chromosomes and conspicuous bands on the short arms of one chromosome pair, where nucleolar organizer regions (NORs) were evidenced by silver-staining. Without CB pretreatment, the NORs were stained conspicuously with CMA₃, but not with DAPI. The latter uniformly stained all chromosomes, but leaving the NORs pale. Combination of CMA₃ or DAPI staining with CB showed distinctive fluorescent blocks in the NOR-bearing short arms of the single chromosome pair along with several bright fluorescent signals on other chromosomes, which were not evidenced by single CMA₃ or DAPI staining. These results suggest a modification of chromatin structure by CB treatment, which may increase the stainability of CMA₃ and DAPI.     

Chromosome complement and nucleolar organization in Podophyllum hexandrum Royle

The karyomorphology of Podophyllum hexandrum Royle (2n=2x=12) is described for the first time. The haploid set comprises one metacentric, three sub-metacentric and two acrocentric chromosomes. Metacentrics are the longest and acrocentrics the smallest in the complement. Secondary constrictions are located in the short arm of two meta-/submetacentrics and in the long arm of the two acrocentric chromosomes. The number of NORs is in agreement with the number of nucleoli organized per root tip nucleus.     

Active nucleolar-organizer regions in polyploid populations of Odontophrynus americanus (Amphibia,Anura) from South Brasil

Specimens of O. americanus collected in eight localities from Rio Grande do Sul and four from Paraná (Brasil) were studied. Patterns of chromosomal localization and activity of nucleolar organizer regions (NORs) were established using the silver banding technique. It was shown that diploid and tetraploid specimens from all localities have homologues of group 4 bearing active NORs. In addition, some populations showed NOR activity on only one homologue number 5 (Uruguaiana, Prudentópolis), 8 (Itaqui) or 2 (Pato Branco). Intrapopulation polymorphism was found, some specimens showing the extra band and others not. Most cells analysed from the intestinal epithelium of an individual showed the same particular staining pattern of NORs, which were unequally stained, suggesting differences in their activity. The tetraploid genomes showed a maximum of five and a minimum of two active NORs. Diploid genomes showed two active NORs and four in the case of one specimen from Pato Branco. Four diploid populations and sympatry of 2n and 4n animals in Santa Bárbara do Sul (RS) were detected for the first time in South Brasil. Karyological evolution, as well as the meaning of number and unequal activity of NORs on tetraploid genomes are discussed.     

Karyotype analysis of Eucinostomus argenteus, E. gula, E. harengulus, and Eugerres plumieri (Teleostei, Gerreidae) from Florida and Puerto Rico

The karyotypes of four gerreids of the western Atlantic Ocean are documented. A diploid chromosome complement of 48 telocentric chromosomes was found in the four species (2N=48t, fundamental number FN=48). No differences were detected either in the number of chromosomes of the standard karyotype, in their karyotype size, or between the karyotypes derived from male or female specimens of any of the species. Chromosome length decreased progressively and slightly from pair 1 to pair 24. The Ag–NOR karyotypes of E. argenteus and E. harengulus were characterized by the position of the nucleolar organizer regions next to the centromere in chromosome pair 1, whereas in E. gula and E. plumieri Ag–NORs were detected in pair 4. The other 46 chromosomes showed a light staining of the centromere with no terminal or intermediate heterochromatic blocks. All Eucinostomus species showed Ag–NORs of similar size, while Eugerres plumieri showed Ag–NORs 10–20% larger than Eucinostomus species. A combination of size and position of the Ag–NORs identified E. gula, while size alone identified E. plumieri. However, the ancestral state for size and position of Ag–NORs could not be established. There was no differential staining of the chromosomes by G-banding. The karyotype of the gerreids appears similar to the hypothetical ancestral karyotype of fish. The phylogenetic relationships among these species could not be established because of the lack of chromosome G-bands. Most likely this indicates a homogeneous distribution of GC nucleotides in the chromosomes.