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Chromosome painting in plants: history and future perspectives.

Chromosome painting was developed in mammalian species nearly four decades ago and rapidly became a powerful tool for chromosome identification, comparative cytogenetics, and evolutionary genome analysis. Comparative chromosome painting among diverse mammals generated much of the foundational knowledge of chromosome structure, chromosomal rearrangements, and karyotype evolution before the advent of whole-genome sequencing. Although chromosome painting was first demonstrated in plants in 2001, its applications remained largely restricted to a few plant lineages until the development of oligonucleotide (oligo)-based chromosome painting in 2015. During the last decade, oligo-based chromosome painting has transformed plant cytogenetics, enabling many investigations that were previously impossible. These studies have provided new insights into meiotic chromosome pairing, crossover formation, chromosome fusion, karyotype stability, and chromosome evolution across diverse plant lineages. This review summarizes the history of technological development of chromosome painting in plants, highlights major discoveries enabled by oligo-based chromosome painting, and discusses future opportunities, particularly the integration of chromosome painting with three-dimensional chromosome and genome biology.

Chromosome Painting

Highly Contiguous Is Not Chromosomally Accurate: Integrated Cytogenetic and Genomic Mapping in Two Turtle Genome.

High-quality genome assemblies are essential for robust research across biological and medical fields. Assembly errors can have far-reaching consequences for downstream analyses, including gene annotation and the inference of synteny. In contrast to the rapid growth of genomic data volume, there is a notable lag in the integration of chromosome-level assemblies with cytogenetic data. We conducted the first direct genome-to-genome comparison, integrating comparative chromosome painting, the alignment of chromosome-specific probes to available genome assemblies, and synteny-based comparison of independent chromosome-level assemblies of the loggerhead sea turtle (Caretta caretta, 2n = 56) and the red-eared slider (Trachemys scripta elegans, 2n = 50). Using two independent sets of flow-sorted chromosome-specific probes in cross-species hybridizations, together with the sequencing and mapping of chromosome-derived DNA libraries, we assigned assembled scaffolds to all physical chromosomes of both species. In C. caretta, chromosomal assignments and genome-wide synteny were fully consistent with the published assembly, except for the reduced sizes of two microchromosome scaffolds, which we attribute to under-representation of repetitive DNA. In contrast, in T. s. elegans, cytogenetic validation of the assemblies revealed a false rearrangement compared to a missed one. Our results show that even highly contiguous vertebrate genome assemblies can misrepresent chromosome structure. When cytogenetic analyses reveal such inaccuracies, updated reference genomes should be generated for widely studied species to enable accurate inference of karyotype evolution and downstream comparative genomic analyses.

FISH

Chromosomal repatterning in Acrididae.

Studies on the chromosomes of the acridid grasshoppers Acrida turrita, Poekilocerus pictus and Chrotogonus oxypterus have led the authors to surmise that structural re-arrangements must have played a major role in chromosomal repatterning and karyotypic evolution. Moreover, the telocentricity noticed in the Cryptosacci was evident in the Chasmosacci without the presence of the metacentric chromosomes to account for the reduction in the chromosome number. Possible trends in the evolution are discussed.

Animals

Comparative analysis of karyotypes in European shrew species. I. The sibling species Sorex araneus and S. gemellus: Q-bands, G-bands, and position of NORs.

The karyotypes of two closely related species of the genus Sorex (Mammalia, Insectivora) were compared with each other by G- and Q-banding techniques and by Ag-AS staining (GOODPASTURE and BLOOM, 1975). By comparing the G-banded karyotypes, it could be ascertained that the basic differences in karyotype between the two species lie in three pericentric inversions, three paracentric inversions, and one reciprocal translocation. This is in near agreement with FORD and HAMERTON (1970), who assumed that both species differ by three pericentric inversions and one tandem translocation. Furthermore, the karyotype of S. araneus (race C) presented by HALKKA et al. (1974) has been compared with the S. araneus of the present report. Considering the species with respect to karyotypic evolution, it is supposed that S. araneus and S. gemellus derive from a common ancestor.

Animals

Zaprionus tuberculatus: chromosome map and gene mapping by DNA in situ hybridization.

The genus Drosophila has long been used as a model of karyotype evolution, demonstrating change by paracentric inversion and occasional centric fusion of an ancestral karyotype of five rod-shaped and one "dot" chromosome. This study shows, by mapping D. melanogaster probes hybridized to polytene chromosomes of Zaprionus tuberculatus, that this ancestral pattern extends beyond the genus Drosophila. A formal polytene chromosome map of Z. tuberculatus is presented.

Animals

[Chromosome complements of eleven bird species of the orders columbiformes, passeriformes and tinamiformes (author's transl)].

The chromosome complements of 11 species of birds from the Columbiformes, Passeriformes and Tinamiformes Orders are described. Their karyotypes' evolution are discussed and pericentric inversions, centric fissions and/or fusions seem to have been of great importance. Banding patterns in the several Orders will be of great importance to know better the mechanisms of chromosomal evolution.

Animals

Unraveling evolutionary relationships in the Sida generic alliance (Malvaceae, Malvoideae): a phylogenetic and cytotaxonomic overview.

Sida (Malvaceae), the largest Malveae-Abutilinae member, has poorly defined morphological limits which overlaps with 11 phylogenetically closely related genera that comprises the "Sida generic alliance". The 12 genera are distributed in the tropics especially in Brazil where one third of its species diversity is found. Evolutionary relationships within Sida generic alliance remain unresolved due to morphological convergence, limited taxon sampling, and lack of integrative approaches including cytogenetic data. We reconstructed the phylogeny of Sida and allied genera using a multilocus dataset (nuclear ITS and seven plastid loci) including 193 species classified in 19 genera and analyzed chromosome evolution using cytogenetic data (chromosome number) for 79 species of the 19 genera. The phylogeny recovered seven clades-Abutilon, Bakeridesia, Callianthe, Gaya, and three Sida clades (I-III)-and confirmed the polyphyly of Sida, the largest genera. We detected reticulate evolution, with incongruence between nuclear and plastid topologies. Chromosome number ranged from 2n = 12 to 60 and represented synapomorphies for most clades. Ancestral character reconstruction indicated that ascending dysploidy and polyploidy predominated in karyotype evolution of Sida and allied genera. Our results reveal taxonomic incongruence in current classifications probably related to reticulate evolution. A generic-level taxonomic revision is necessary and should rely on integrated phylogenetic and karyotypic evidence. This study provides a framework for phylogenetic systematics and emphasizes the role of Brazil as a hotspot for plant genomic research.

Phylogeny

Translocations of acrocentric chromosomes and their implications in the evolution of sheep (Ovis).

Cytogenetic evidence suggests that the caprids (sheep and goats) evolved from a common ancestor with a 2n=60 karyotype. Although goats (Capra) retained the primitive 2n=60 karyotype, sheep (Ovis) underwent a sequential reduction in the number of chromosomes by means of acrocentric translocation. The formation of the first metacentric autosome (M1) occurred in the aoudad (Ammotragus) and urial (O. vignei), resulting in a 2n=58 karyotype. The G-bands are homologous, which implies both genotypes arose from a common ancestor, possibly a rupicaprid. Based on G-bands, acrocentric chromosomes 1 and 7 of the 2n=60 karyotype formed the M1. The X chromosome, which is the second longest acrocentric in the 2n=60 karyotype, became the longest acrocentric in Ammotragus and Ovis (2n=58). The second pair of metacentrics to evolve, which is ranked in the M3 position of the 2n=54 karotype, resulted from the translocations of acrocentric chromosomes 4 and 14 or 15 in the 2n=60 karyotype. The M2 was the third pair of metacentrics to be formed and resulted from the translocations of acrocentric chromosomes 3 and 12 or 13 in the 2n=60 karyotype. The G-bands of all 2n=54 karyotypes are homologous, which indicates origin from a common ancestor. Evidence is presented that suggests a prezygotic selection is bringing about a reduction in diploid chromosome numbers. The possible roles of fission and fusion in the karyotypic evolution of Ovis are discussed.

Animals

Benign mixed tumor of canine mammary gland showing an r(X) and trisomy 5 as the only clonal abnormalities.

We report a benign mixed tumor of the canine mammary gland which showed an r(X) and trisomy 5 as the only clonal karyotypic deviations. Clonal aberrations were observed in 79 of 160 of the metaphases. Of these, 48 cells had both the r(X) and trisomy 5, whereas the remaining metaphases were characterized by the r(X) as the only clonal aberration. We conclude that formation of the ring chromosome was the first abnormality, followed by trisomy 5 during the course of karyotypic evolution.

Animals

Contributions to the karyology of Euconcocephalus incertus and Allodapia aliena (Tettigoniidae, Orthoptera.

The chromosome complement of two species of Indian Tettigoniids, namely, Euconcocephalus incertus (2n male = 20A + XX) and Allodapia aliena (2n female = 28A + XX) have been reported for the first time. The former species has two pairs of metacentric and two pairs of submetacentric autosomes, and a pair of metacentric sex chromosomes, whereas in the latter species all the chromosomes are telocentric and the sex chromosomes are very long. The probable occurrence of centric fusions, translocations and inversions during the karyotypic evolution of these grasshoppers is discussed.

Animals

Clonal evolution of marker chromosomes in a case of myelofibrosis with myeloid metaplasia and myeloblastic transformation.

The diverse spectrum of acquired chromosome abnormalities in a female patient with myelofibrosis and myeloid metaplasia is described. A sequence of karyotypic evolution involving a ring chromosome is postulated. The terminal clinical picture was unusual in that there was obstructive renal failure from extramedullary myeloblastic transformation and infiltration of the bladder, and this was also present in other sites. Initially neutrophils showed low alkaline phosphatases activity but latterly two distinct populations in which cells had either high activity or none.

Alkaline Phosphatase

Mauritius type black rats with peculiar karyotypes derived from Robertsonian fission of small metacentrics.

All seventeen black rats collected from Mauritius Island were characterized by having many extra small acrocentric autosomes. Their basic karyotype was of Oceanian type, because of the presence of the large metacentric M1 and M2 pairs, but chromosome numbers in 13 specimens among them were 42, those of 3 specimens 43, and those of the remaining one specimen 44. Although the Oceanian type rat had 2 small acrocentric autosomes (pair no. 13), 16 Mauritius rats had 10 small acrocentrics, and the remaining one had 8 small acrocentrics. Comparative karyotype analysis between Oceanian and Mauritius type rats showed that the extra small acrocentrics found in Mauritius rats were due to Robertsonian fission of small metacentric pairs no. 14 and 18 of the original Oceanian type rat. Only one rat with 8 small acrocentrics showed the heteromorphic pair no. 18 consisting of one metacentric and two acrocentrics. The large metacentric M1 chromosome in 13 of 17 rats examined showed homologous pair, but two of them were heteromorphic by involving one metacentric M1 and two acrocentrics. In the remaining two rats M1 chromosome was not observed, but acrocentric pairs no. 4 and 7 were included. These acrocentrics were also suggested to be originated from Robertsonian fission of the large metacentric M1 chromosome. Robertsonian fission seemed to be one of the important mechanism found in karyotype evolution.

Animals

Variation of C-bands in the chromosomes of several subspecies of Rattus rattus.

All subspecies of black rats (Rattus rattus) used in the present study are characterized by having large and clear C-bands at the centromeric region. The appearance of the bands, however, is different in the subspecies. Chromosome pair No. 1 in Asian type black rats (2n=42), which are characterized by an acrocentric and subtelocentric polymorphism, showed C-band polymorphism. In Phillipine rats (R. rattus mindanensis) the pair was subtelocentric with C-bands, but in Malayan black rats (R. rattus diardii) it was usually acrocentric with C-bands. In Hong-Kong (R. rattus flavipectus) and Japanese black rats (R. rattus tanezumi) it was polymorphic with respect to the presence of acrocentrics with C-bands or subtelocentrics without C-bands. The other chromosomes pairs showed clear C-bands, but in Hong-Kong black rats the pairs No. 2 and 5 were polymorphic with and without C-bands. In Japanese black rats, 6 chromosome pairs (No. 3, 4, 7, 9, 11 and 13) were polymorphic in regard to presence and absence of C-bands, but the other 5 chromosome pairs (No. 2, 5, 6, 8 and 10) showed always absence of C-bands. Only pair No. 12 usually showed C-bands. C-bands in small metacentric pairs (No. 14 to 20) in Asian type black rats generally large in size, but those in the Oceanian (2n=38) and Ceylon type black rats (2n=40) were small. In the hybrids between Asian and Oceanian type rats, heteromorphic C-bands, one large and the other small, were observed. Based on the consideration of karyotype evolution in the black rats, the C-band is suggested to have a tendency toward the diminution as far as the related species are concerned.

Animals

Origin and significance of centric fusions in domestic sheep.

The karyotypes of 731 sheep of various breeds were studied and considered in association with previous chromosome studies of domestic sheep. A high incidence of the t2 trnslocation was found in two pedigree flocks of New Zealand Romney sheep. One of these flocks was established over 100 years ago and it is suggested that this translocation originated in the Romney Marsh breed of sheep in England. A naturally occurring double translocation heterozygote 52t1t2 was reported for the first time. A further flock of sheep of the Perendale breed was found with a high incidence of dicentric chromosome fusion that was identified as the t3 translocation. The apparently common occurrence of chromosome polymorphism, due to centric fusions, in domestic sheep is discussed in relation to karyotype evolution among both domestic and wild sheep.

Animals

Sequential analysis of transplantable hepatocellular carcinomas.

In a previous study multiple characteristics of chemically induced primary hepatocellular carcinomas were described and examined during the initial transplant generations. The present communication reports on these characteristics in subsequent transplant generations followed over a 2-year period. In almost all instances the growth rate, morphology, chromosome composition, and plasma protein and alpha-fetoprotein synthesis of individual tumors have remained relatively constant. However, one spontaneous subline of a diploid tumor demonstrated a sudden extensive rearrangement of its chromosomes simultaneous with a significant acceleration of growth rate. Despite karyotypic evolution, it retained the functional characteristics of diploid tumors, producing no plasma protein or alpha-fetoprotein.

2-Acetylaminofluorene

Karyotypic abnormalities in transformed chronic granylocytic leukaemia.

Chromosomal abnormalities in three cases of chronic granylocytic leukaemia are presented. The relative importance of 'specific' chromosomal abnormalities, additional to the Ph1 in the karyotypic evolution of chronic granylocytic leukaemia is discussed. A new abnormal metacentric chromosome is described.

Adult

Karyologic studies on hybrids between Asian, Ceylonese, and Oceanian type black rats, with a note on an XO female occuring in the F2 generation.

F1 hybrids between the Asian type (2n=42) and Ceylonese type black rats (2n=40) invariably showed 41 chromosomes, while those between the Oceanian type (2n=38) and the Ceylonese type usually showed 39 chromosomes. Twelve F2 hybrids (three litters) were obtained only from the latter combination. Among them, the karyotypes of nine rats were successfully analyzed. They were segregated into the Oceanian type (three rats) and the F1 type (six rats). No Ceylonese type was obtained. From the results of fertility testing in F1 hybrids between these three variants, the Oceanian type is assumed to be more closely related to the Ceylonese type than the Asian type. This coincided well with the results of karyotypic evolution of these geographic variants. Among the nine F2 hybrids examined, one female consistently showed 37 chromosomes with one X chromosome. This XO female appeared to be a normal female and to have been a sporadic natural occurrence.

Animals

A hypodiploid clone and its duplicate in acute lymphoblastic leukemia.

Examination of the bone marrow of a 63-year-old man who had acute lymphoblastic leukemia revealed a population of cells with 32 chromosomes and another population with 64 chromosomes, the karyotypical exact duplicate of the first clone. The karyotypic evolution was studied and the findings compared with those described in two similar cases previously reported. It is postulated that severe hypodiploidy is associated with reduced capability of cellular survival, promoting a strong tendency for duplication.

Bone Marrow