The reannealing kinetics of denatured DNA fragments from 23 species of higher plants have been studied, using hydroxylapatite chromatography to distinguish reannealed from single-stranded DNA. The 2C nuclear DNA contents of the species varied between 1.7 and 98 pg. The proportions of DNA in species with a nuclear DNA mass above 5 pg that reannealed with the kinetics of sequences present in more than 100 copies were high (69–92% with a mean of 80±2.0%). For species with less than 4 pg of DNA, the mean proportion of repeated-sequence DNA was 62±2.9%. It is concluded that most of the variation in nuclear DNA mass in higher plant chromosomes can be accounted for by variation in repeated-sequence DNA. The consequences of altering the adapted DNA content of a species by the addition of families of repeated sequences are discussed in relation to the proportion of repeated-sequence DNA.

Genome size and the proportion of repeated nucleotide sequence DNA in plants.

Highly differentiated heteromorphic sex chromosomes were found in the karyotypes of Pyxicephalus adspersus (= Rana adspersa) and analysed with the various banding methods. The W chromosome is considerably smaller than the Z chromosome and consists to a great proportion of constitutive heterochromatin. The chromosome lengths and the DNA content of this species have the lowest values determined in the Ranidae to date. The nucleolus organizer regions in the chromosomes reveal a high frequency of inter-individual variations (duplications, triplications) and aberrations. In one female animal, a translocation between the W chromosome and a nucleolus organizer was identified. Preliminary results indicate, that the same chromosome pair which in P. adspersus constitutes the heteromorphic sex chromosomes is still in an initial stage of morphological differentiation in P. delalandii. Although the ZZ/ZW sex chromosomes in Pyxicephalus have developed independently of the ZZ/ZW sex chromosomes in birds and reptiles, they seemed to have passed through identical stages of morphological differentiation. In this process, the heterochromatinization of the W chromosome had a decisive function.

Chromosome banding in Amphibia

A comparison of the ribosomal DNA's of Xenopus laevis and Xenopus mulleri: the evolution of tandem genes☆

Composition and structure of chromosomal and amplified ribosomal DNA's of Xenopus laevis

Publisher Summary This chapter reviews that the ribosomal cistrons of both bacteria and eukaryotes, with their unique and well-defined characteristics, have increasingly attracted the attention of biochemists and geneticists. It discusses that the arrangement of the ribosomal cistrons on the chromosomal DNA, and they have been mapped in some detail by means of biophysical techniques. Ribosoinal cistrons were the first Mendelian genes isolated in pure form free from other DNA and it has now become possible to investigate their organization and composition and to study their biological activity in vitro . The chapter focuses on the organizational features and primary structure of DNA. It presents a picture of the detailed physical structure of the ribosomal cistron and how it is integrated into the genomic DNA. It has considered the divergence of ribosomes in different species and the variation of base sequence within the ribosomal cistrons of a single organism, both of which have important implications for cell differentiation and for evolutionary theory.

The ribosomal RNA cistrons.

In metaphase preparations from leucocytes of the toad, Bufo marinus, conspicuous secondary constrictions are present in the number 7 pair of chromosomes. These constrictions were considered to be the nucleolar organizers since they were associated with nucleoli during prophase. In 35 out of 60 individuals taken from natural populations, the homologous nucleolar organizers produced two equal-sized nucleoli and secondary constrictions (Group I animals). Pour animals (Group II) had only one very large secondary constriction in the majority of their metaphase preparations and an abnormally high frequency of cells containing one nucleolus. The remaining 21 animals (Group III) had unequal-sized constrictions in most of their metaphases but were more variable than the individuals of Groups I or II since they also had metaphases with two equal constrictions or only one constriction. The DNA from individuals of each group was hybridized with radioactive ribosomal RNA in order to correlate the size of nucleoli and constrictions with the amount of DNA (rDNA) homologous to ribosomal RNA. The two animals of Group II which were studied contained 0.056% of their genome homologous to ribosomal RNA a value considerably higher than those found for any of the animals of Groups I or III. These high values for rDNA coupled with the morphological appearance of the nucleolar homologues suggested a duplication of the nucleolar organizer in the homologue with the long constriction. The amount of rDNA in animals of Group I and III varied between 0.025 and 0.048% of the genome. Although the animals with unequal-sized constrictions (Group III) had generally lower contents of rDNA than those with equal-sized constrictions (Group I), the values overlapped between the two groups. Further evidence which correlates the size of nucleoli with the number of ribosomal RNA genes comes from studies with a “small nucleolar” mutant of the Mexican axolotl (Ambystoma mexicanum). Animals homozygous for this deletion were found to contain only 55% of the complement of rDNA present in the wild type. It is concluded that partial deletions and duplications of the nucleolar organizer as well as highly variable contents of rDNA are common in the genome of these amphibians.

Donald D. Brown

Papers

Variation in the activity of nucleolar organizers and their ribosomal gene content.