Interspecific interactions between three sympatric species of kangaroo rats (Dipodomys)

Interspecific interactions between Dipodomys merriami, D. ordii, and D. panamintinus were observed in the laboratory. The largest species, D. panamintinus, was the most dominant while the smallest species, D. merriami, was the least dominant. All interactions were characterized by extreme fighting. Laboratory and field evidence indicate that interspecific aggression in the field may be one mechanism keeping these sympatric species of kangaroo rats ecologically separate.     

Location of satellite DNAs in the chromosomes of the kangaroo rat (Dipodomys ordii)

The location of satellite DNA sequences in metaphase chromosomes has been studied in the kangaroo rat by the in situ hybridization technique, staining techniques and phase contrast microscopy. The HS- satellite DNA is located at the kinetochores of all but three chromosome pairs. The HD satellite is located predominantly in the short arms of the chromosomes containing HS- and in the kinetochores of chromosome pairs that lack HS-. The regions that contain the satellite DNA sequences can also be identified by the Giemsa staining technique, and can be visualized with phase contrast microscopy or following Feulgen staining of fixed chromosome preparations.     

Comparative analysis of different satellite DNAs in four Mytilus species.

We report the characterization of three satellite DNAs in four species of mussel: Mytilus edulis, Mytilus galloprovincialis, Mytilus trossulus, and Mytilus californianus. The monomers of the Apa I satellite DNAs were 173, 161, and 166 bp long. These satellite monomers were used to construct phylogenetic trees to infer relationships among these species. The topologies obtained clearly indicate that M. californianus is the most divergent species with respect to the other three. Furthermore, localization of satellite DNAs on metaphase chromosomes was performed using fluorescent in situ hybridization (FISH). Fluorescent signals revealed a different organization and distribution of these three satellite DNAs.     

Isolation of twelve satellite DNAs from Drosophila hydei

The genome of a Drosophila hydei genotype with a reduced amount of heterochromatin was fractionated by three cycles of preparative gradients: firstly in Ag⁺/Cs₂SO₄, secondly in actimomycin D/CsCl, and finally in neutral CsCl. Using this method, twelve highly repetitive simple-sequence satellites were isolated. Ten of them comprised only a minor amount of the genome in contrast to the two major satellites found earlier¹ (p = 1.696 and 1,714 g/cm³). These minor satellites were characterized by their banding in the gradient systems used, by their density in neutral CsCl, and by their melting point. Using these characteristics, it was found that the fractions of the Ag^?/Cs₂SO₄ gradient do not contain purified single components, because up to five different satellites band in the same position of the Ag^?/Cs₂SO₄ gradient. It was possible to isolate a high number of satellites even from a genome with a reduced amount of heterochromatin. Thus, the D hydei heterochromatin does not domain one unique highly repetitive sequence DNA, but is comprised of many different satellite sequences.     

The non-regular orbit: three satellite DNAs in Drosophila martensis (buzzatii complex, repleta group) followed three different evolutionary pathways

The genome of species from the buzzatii cluster (buzzatii complex, repleta group) is hosted by a number of satellite DNAs (satDNAs) showing contrasting structural characteristics, genomic organization and evolution, such as pBuM-alpha (~190 bp repeats), pBuM-alpha/beta (~370 bp repeats) and the DBC-150 (~150 bp repeats). In the present study, we aimed to investigate the evolution of these three satDNAs by looking for homologous sequences in the genome of the closest outgroup species: Drosophila martensis (buzzatii complex). After PCR, we isolated and sequenced 9 alpha, 8 alpha/beta and 11 DBC-150 sequences from this species. The results were compared to all pBuM and DBC-150 sequences available in literature. After D. martensis split from the buzzatii cluster some 6 Mya, the three satDNAs evolved differently in the genome of D. martensis by: (1) maintenance of a collection of major types of ancestral repeats in the genome (alpha); (2) fixation for a single major type of ancestral repeats (alpha/beta) or (3) fixation for new divergent species-specific repeat types (DBC-150). Curiously, D. seriema and D. martensis, although belonging to different and allopatric clusters, became independently fixed for the same major type of alpha/beta ancestral repeats, illustrating a rare case of parallelism in satDNA evolution. The contrasting pictures illustrate the diversity of evolutionary pathways a satDNA can follow, defining a “non-regular orbit” with outcomes difficult to predict.     

An analysis of sandbathing and grooming in the kangaroo rat (Dipodomys merriami)

The sandbathing and grooming behaviour of ten kangaroo rats (Dipodomys merriami) were recorded on sand and woodchip substrates after periods of 0,1,5 and 10 days without sand. Sandbathing is restricted to the sandy substrate. Grooming occurs on both, but with a higher frequency on sand. Increases in both grooming and sandbathing occur with increasing sand deprivation, but the temporal patterning does not change. D. merriami tends to alternate sandbathing components in contrast to other Dipodomys species. Lipid on the pelage increases noticeably with sand deprivation and decreases during a sandbathing bout; sand appears to be removed from the pelage by shaking and grooming. These findings suggest a three-process system for care of the body surface.     

Restriction endonuclease patterns of three baculoviruses isolated from species of Heliothis

The DNA of three baculoviruses propagated in larvae of a common host species (Heliothis zea) were easily distinguished from each other by their restriction endonuclease patterns. Molecular weights of 79.7 ± 7.3, 119.6 ± 5.1, and 86.6 ± 6.3 × 10⁶ daltons were estimated for the viral genome of a single-embedded nucleopolyhedrosis virus isolated from Heliothis zea, a multiple-embedded nucleopolyhedrosis virus isolated from Heliothis armigera, and a granulosis virus isolated from Heliothis armigera, respectively.     

Chromosomal distribution and organization of three cervid satellite DNAs in Chinese water deer (Hydropotes inermis)

The species-specific profile and centromeric heterochromatin localization of satellite DNA in mammalian genomes imply that satellite DNA may play an important role in mammalian karyotype evolution and speciation. A satellite III DNA family, CCsatIII was thought to be specific to roe deer (Capreolus capreolus). In this study, however, this satellite DNA family was found also to exist in Chinese water deer (Hydropotes inermis) by PCR-Southern screening. A satellite III DNA element of this species was then generated from PCR-cloning by amplifying this satellite element using primer sequences from the roe deer satellite III clone (CCsatIII). The newly generated satellite III DNA along with previously obtained satellite I and II DNA clones were used as probes for FISH studies to investigate the genomic distribution and organization of these three satellite DNA families in centromeric heterochromatin regions of Chinese water deer chromosomes. Satellite I and II DNA were observed in the pericentric/centric regions of all chromosomes, whereas satellite III was distributed on 38 out of 70 chromosomes. The distribution and orientation of satellite DNAs I, II and III in the centromeric heterochromatin regions of the genome were further classified into four different types. The existence of a Capreolus-like satellite III in Chinese water deer implies that satellite III is not specific to the genus Capreolus (Buntjer et al., 1998) and supports the molecular phylogeny classification of Randi et al. (1998) which suggests that Chinese water deer and roe deer are closely related.     

Analysis of in vitro replication of different DNAs.

The conversion of single-stranded circular DNA to duplex DNA in vitro occurs by at least three different mechanisms. These differences reside in the manner of priming of these DNAs. In contrast, the elongation of primed DNA templates is a general reaction. A number of these proteins have been isolated and further characterized. In addition, cell-free preparations capable of supporting phi X RFI DNA replication as well as the synthesis of progeny viral phi X174 single-stranded circular DNA have been prepared.     

Characterization of heterochromatin in different species of the Scilla siberica group (Liliaceae) by in situ hybridization of satellite DNAs and fluorochrome banding

Satellite DNAs have been isolated from the monocotyledonous plants Scilla siberica, S. amoena, S. ingridae (all are highly GC-rich), and S. mischtschenkoana by using the Ag⁺ –Cs₂SO₄ density centrifugation technique. Hybridization in situ has been performed with ₃H-cRNA to these satellite DNAs in all four species. In each species, the endogenous satellite DNA is located mainly in intercalary and major heterochromatin bands associated with terminal regions and nucleolar organizer regions (NORs) but not in centromeric regions. Patterns observed after cross-species hybridization show a high degree of satellite DNA homology between S. siberica, S. amoena, and S. ingridae. By contrast, satellite DNA of S. mischtschenkoana consists largely of different, non homologous DNA sequences, with two exceptions: (i) the NORs of all four species contain similar satellite sequences, and (ii) a strong homology exists between the satellite DNA of S. mischtschenkoana and centromeric DNA of S. siberica but not with those of S. amoena and S. ingridae. — Heterochromatin has also been characterized by the AT-specific fluorochromes quinacrine (Q) and DAPI and the GC-specific agent chromomycin A₃ (CMA₃), in combination with two counterstaining techniques. While CMA₃-fluorescence is largely in agreement with data on base composition and location of the specific satellite DNAs, the results with Q and DAPI are conflicting. Prolonged fixation has been found to change the fluorescence character in certain instances, indicating that other factors than the base sequence of the DNA also play a role in fluorochrome staining of chromosomes. The results are discussed in relation to the taxonomy and phylogeny of the four species.     

Daily cycles of hibernation in the kangaroo rat, Dipodomys merriami

Thirty-six kangaroo rats were captured in the Mojave Desert near Las Vegas. Eight rats, group A, were kept at room temperature, 25 ± 2 °C. Ten, Group B, were kept at 13–15 °C and fed ad libitum. Eighteen, group C, were kept at 13–15 °C and fed only 2 g of dry oats daily. Rate of O₂ consumption, VO₂, food intake, rectal temperature, T_re., body weight and composition were measured in all groups. Group B showed a significant increase in VO₂, and food intake and no change in T_re body weight as compared to group A. After 2 days on the limited food intake group C began to lose weight, and T_re then began to fluctuate ranging between 14 and 36 °C; the animals exhibited hibernation when the T_re was low. Eight of the 18 rats of group C reached T_re values of 14 °C or below; only one of these survived. The lowest T_re in the other 10 was 15 °C; all survived. Chemical analysis of the homogenized rats showed a significant decrease in body fat in group C to average 1.0% contrasted with 7.8% in group B and 3.7% in group A. The VO₂ ranged from 1.9 ml/g hr at a T_re of 36.5 to 0.2 ml/g hr at a T_re of 15 °C. In conclusion D. merriami utilizes hibernation as an effective adaptive mechanism to carry out their essential body functions when on a limited food intake. It seems that maintaining a level of more than 1.5% of body fat is essential for successful arousal from hibernation.     

Moon-related surface activity of bannertail (Dipodomys spectabilis) and fresno (D. Nitratoides) kangaroo rats

Temporal aspects of nocturnal activity of bannertail kangaroo rats were measured in the field. Maximum activity was about twenty minutes aftyer sunset, thereafter declining throughout the night. Activity was about three times greater when the moon was down than when it was up. Activity shifted somewhat from the open to vegetation cover when the moon was up. Laboratory measures of activity from captive bannertails were also obtained. The activity of bannertails kept in small pens resambled corresponding aspects of the field data. Activity in wheels was unlike activity in the field, and the effects of a laboratory ‘moon’ were opposite to those of the real moon upon bannertails in the field.     

Isoenzyme variation patterns and species concept in Astragalus gossypinus and Astragalus persicus complexes (Fabaceae) in Iran

Isoenzyme electrophoresis was employed to examine the relationships of 21 individuals representing four populations of Astragalus gossypinus complex as well as 20 individuals representing five populations of Astragalus persicus complex. A total of 27 bands from three enzyme systems (superoxide dismutase, esterase, and peroxidase) were obtained. UPGMA clustering method resulted in two distinct clusters for different populations corresponding to the two species complexes analysed. In both species, populations distributed on Alborz mountain range in northern Iran form separated clusters from those distributed on Zagros mountain range in the West. The results also show that both species complexes exhibit a high diversity based on Shannon–Weaver diversity index (H′) compared with other species of Astragalus reported previously. The mean number of bands per each presumed isoenzyme ranges from 2.14 to 3.57. The value of Euclidean distance ranges from 1.251 to 3.152. Our data suggest that both species should be circumscribed wider than that treated by most taxonomists, and several taxonomic names should be reduced under synonymy of the corresponding species.     

Ovule morphology, embryogenesis and seed development in three Hylocereus species (Cactaceae)

Pre-embryonic and embryonic stages and seed developments were studied in the diploids Hylocereus monacanthus and Hylocereus undatus and the tetraploid Hylocereus megalanthus. Ovule morphology was similar among species except for micropyle entrance. H. monacanthus had the thickest and most robust suspensor. Embryo developmental time, measured from fertilization to maturity, was significantly more prolonged in H. megalanthus. Typical to Cactaceae, the seed coat was formed by one layer of sclerenchymatous cells, but was more lignified in H. megalanthus. Morphological features common to all species included (1) cellular type endosperm with independent patterns of development in the chalazal and micropylar zones, forming a haustorium layer from the chalazal zone to the embryo; (2) an endothelial layer surrounding the embryo sac almost complete; (3) a nucellar summit growing into the micropyle; and (4) a placental obturator and a funicle connecting the ovarian tissue to the ovule. Seed development was typically endospermic (exendospermic orthodox seeds). Anomalies included two egg cells in the same embryo sac, two embryos developing in the same ovule, and embryos developing from the chalazal pole region. Total seed number and seed viability were significantly lower in H. megalanthus than in the other two taxa. Embryos at different developmental stages were observed in aborted H. megalanthus seeds.