Albumin evolution in polyploid species of the genus Xenopus.

Electrophoresis of serum from 21 Xenopus species and subspecies reveals variable numbers of albumin bands. The diploid X. tropicalis has one albumin, while the tetraploid species (laevis, borealis, muelleri, clivii, fraseri, epitropicalis) have two. The octoploid species (amieti, boumbaensis, wittei, vestitus, andrei) have two to three bands, and the dodecaploid X. ruwenzoriensis has three. The molecular weight of the Xenopus albumins varies from 68 kd (in the tropicalis group) to 74 kd. The subspecies of X. laevis possess two albumins of different molecular weights (70 and 74 kd), whereas most species have only 70-kd albumins. Peptide maps have been obtained from albumin electromorphs by limited proteolysis in sodium dodecyl sulfate (SDS) gels, using S. aureus V8 protease. The peptide patterns produced by electromorphs from the same tetraploid Xenopus species generally differ from each other, suggesting that the two albumin genes contain a substantial amount of structural differences. In addition, the peptide maps are diagnostic for most tetraploid species and for some subspecies of X. laevis as well. Proteolysis of albumins from most octoploid and dodecaploid species results in patterns which are very similar to the ones produced by the electromorphs from X. fraseri. The albumins of X. vestitus differ from those of the other octoploid species. X. andrei possesses two fraseri-type and one vestitus-type albumin, which indicates that it probably originated by allopolyploidy.

Evolution of globin expression in the genus Xenopus (Anura: Pipidae).

Comparison of electrophoretic globin phenotypes of 18 different Xenopus taxa reveals four different basic types of banding patterns. Each type includes species that also are similar in their morphological, cytogenetical, and biochemical characteristics. Three of these patterns reflect distinct evolutionary lines, while the fourth may be interpreted as the intersection of two of these lines. The composition of the basic pattern of the highly polyploid species is consistent with an allopolyploid origin of most of these species. The number of distinct globin polypeptides--four in the only extant diploid species, X. tropicalis, and five or more in most of the tetraploid species, including X. laevis--suggests that primordial globin genes had undergone duplication either before or after the tetraploidization event. Finally, the individual globin phenotypes are excellent molecular markers that are of great help in identifying the various species but not the subspecies.

Cellular DNA content in different species of Xenopus.

1. Biochemical methods were applied to determine the DNA content per cell in ten species and subspecies of Xenopus. 2. Cell density and DNA concentration were measured in a suspension of blood cells. 3. DNA was determined by the diphenylamine method and values were compared with those obtained by the ultraviolet absorption method and by cytofluorometry. 4. A good degree of reproducibility was demonstrated by a simple biochemical method which yielded values compatible with cytofluorometry. 5. Results suggested that the DNA of Xenopus muelleri, and perhaps that of other species with a large genome size, may be specially protected from chemical degradation.

A comparison of the karyotype, constitutive heterochromatin, and nucleolar organizer regions of the new tetraploid species Xenopus epitropicalis Fischberg and Picard with those of Xenopus tropicalis Gray (Anura, Pipidae).

A cytological analsis of the recently discovered tetraploid species Xenopus epitropicalis was carried out, using, in addition to the classical orcein method, silver staining and alkaline Giemsa banding techniques. The chromosome number of X. epitropicalis was found to be 40. The chromosomes can be grouped in to sets of four similar chromosomes (quartets), resembling the karyotype of X. tropicalis (2n = 20). However, C-band patterns revealed heterogeneity within the quartets, dividing each of them into two pairs of homologous chromosomes ("duets"). Moreover, there are differences in the position and distribution of constitutive heterochromatin between the karyotypes of X. epitropicalis and X. tropicalis. The secondary constrictions stained by silver and representing the nucleolar organizer regions (NOR's) appear in both species on chromosome pair 5. During meiosis, usually only bivalents appear in X. epitropicalis. The question of whether this species is of autopolyploid or allopolyploid origin cannot be answered with certainty; however, it seems to have a common ancestor with X. tropicalis.

The karyotype of the hexaploid species Xenopus ruwenzoriensis Fischberg and Kobel (Anura: Pipidae).

The chromosome number of the hexaploid species Xenopus ruwenzoriensis is 108. The chromosomes fall into sets of six usually similar chromosomes (hextets) which can be classified into the morphological groups characteristic for the genus Xenopus. However, the three distinct secondary constrictions, typical for X. ruwenzoriensis, appear on two homologs only within a particular hextet, unmasking an underlying heterogeneity. The secondary constrictions on chromosome pair 11 show chromosomal association and are considered to represent the nucleolar organizer regions. There is no evidence of dimorphic sex chromosomes. Plates of spermatocyte metaphase I generally show bivalents, but single large multivalents have also been observed. In the majority of cases, 54 dyads are found in second spermatocyte metaphases. X. ruwenzoriensis is, therefore, either an ancient autohexaploid or, more likely, an allopolyploid of more recent origin.

Two new polyploid Xenopus species from western Uganda.

2 new species of the anuran genus Xenopus have been found in western Uganda: X. ruwenzoriensis sp.n. with the hexaploid chromosome number of 108 in the Semliki Valley, west of the Ruwenzori, and X. species nova with the tetraploid chromosome number of 72 in and near lake Bunyoni.

DNA content in the genus Xenopus.

Nuclear DNA amounts were determined by cytofluorometry for twelve species and subspecies of the genus Xenopus. Absolute values, in pg per nucleus, were obtained by direct comparison with human lymphocyte nuclei. The lowest DNA amount (3.55 pg) was found in X. tropicalis, which possess only 20 chromosomes, and the highest (16.25 pg), in the hexaploid X. ruwenzoriensis, with 108 chromosomes. The two recently discovered tetraploid species, X. sp.n. and X. vestitus have, respectively 12,57 and 12.83 pg of DNA. Among the species and subspecies with 36 chromosomes, the DNA content ranges from 6.35 to 8.45 pg.

Albumin phylogeny for clawed frogs (Xenopus).

Comparisons of albumin indicate that the frogs commonly used by North American molecular and developmental biologists under the name of Xenopus muelleri belong to another species, X. borealis. Phylogenetic analysis of the albumin data reveals two major groups of Xenopus species, one containing only X. tropicalis and the other, called the X. laevis grou, containing the remaining species of the genus. The phylogenetic tree, in conjunction with evidence from chromosomes and DNA content, leads to the hypothesis that total genome duplication occurred in the common ancestor of the X. laevis group.

The karyotype of the tetraploid species Xenopus vestitus Laurent (Anura: pipidae).

Xenopus vestitus possesses 72 chromosomes, wherease in the majority of known Xenopus species there are 2n = 36. During meiosis, 36 bivalents are usually observed at metaphase I and 36 chromosomes at metaphase II. Arranged according to size and centromere position, the chromosomes form the same basic morphologic groups typical of the genus Xenopus. However, the groups are composed of quartets of four similar chromosomes instead of diploid pairs of homologs. The exception to this arrangement involves chromosomes bearing secondary constrictions, which in X. vestitus are represented by two different pairs of homologs, one of which shows, in 39% of the observed mitoses, somatic association and is, therefore, considered to carry the nucleolar organizer. X. vestitus represents either a case of ancient autotetraploidy or, more likely, one of allotetraploidy of more modern origin.