Further insights on chromosomal evolution of the genus Enyalius with karyotype description of Enyalius boulengeri Etheridge, 1969 (Squamata, Leiosauridae)

The genus Enyalius is composed of 10 described species inhabiting forest areas in Amozônia, Cerrado and Atlantic forest biomes. Currently, eight species with high levels of chromosome variation have been karyotyped. The study aims to characterize the karyotype of Enyalius boulengeri, with classical and molecular techniques, and improve knowledge about the karyotype evolution of the lizard genus Enyalius. The species has 2n = 36 chromosomes (8m + 4sm + 24mc), FN = 24; NORs and 18S rDNA were subtelomeric and located on chromosome pair 2. Repetitive DNA probes (CAT)10 accumulated on centromeric and terminal regions of some macrochromosomes. (GA)15 probe showed conspicuous accumulation on the pericentromeric region of chromosome pairs 1 and 6. Repetitive FISH patterns obtained with (GC)15 probe marked the pericentromeric region of the first chromosome pair. All probes showed accumulation in the microchromosomes. The chromosomal formula found on E. boulengeri has been considered the ancestral karyotype for pleurodont Iguania. The genus Enyalius is characterized by two distinctive chromosomal groups; one with highly conserved karyotypes, whereas the other is karyotypically diverse. Our molecular cytogenetics data are promising and will increase knowledge about the genus Enyalius chromosome evolution.

Physical localization of 45S rDNA in Cymbopogon and the analysis of differential distribution of rDNA in homologous chromosomes of Cymbopogon winterianus

Cymbopogon, commonly known as lemon grass, is one of the most important aromatic grasses having therapeutic and medicinal values. FISH signals on somatic chromosome spreads off Cymbopogon species indicated the localization of 45S rDNA on the terminal region of short arms of a chromosome pair. A considerable interspecific variation in the intensity of 45S rDNA hybridization signals was observed in the cultivars of Cymbopogon winterianus and Cymbopogon flexuosus. Furthermore, in all the varieties of C. winterianus namely Bio-13, Manjari and Medini, a differential distribution of 45S rDNA was observed in a heterologous pair of chromosomes 1. The development of C. winterianus var. Manjari through gamma radiation may be responsible for breakage of fragile rDNA site from one of the chromosomes of this heterologous chromosome pair. While, in other two varieties of C. winterianus (Bio-13 and Medini), this variability may be because of evolutionary speciation due to natural cross among two species of Cymbopogon which was fixed through clonal propagation. However, in both the situations these changes were fixed by vegetative method of propagation which is general mode of reproduction in the case of C. winterianus.

Characterization of the Durum Wheat-Aegilops tauschii 4D(4B) Disomic Substitution Line YL-443 With Superior Characteristics of High Yielding and Stripe Rust Resistance

Durum wheat is one of the important food and cash crops. The main goals in current breeding programs are improving its low yield potential, kernel characteristics, and lack of resistance or tolerance to some biotic and abiotic stresses. In this study, a nascent synthesized hexaploid wheat Lanmai/AT23 is used as the female parent in crosses with its AB genome donor Lanmai. A tetraploid line YL-443 with supernumerary spikelets and high resistance to stripe rust was selected out from the pentaploid F7 progeny. Somatic analysis using multicolor fluorescence in situ hybridization (mc-FISH) revealed that this line is a disomic substitution line with the 4B chromosome pair of Lanmai replaced by the 4D chromosome pair of Aegilops tauschii AT23. Comparing with Lanmai, YL-443 shows an increase in the number of spikelets and florets per spike by 36.3 and 75.9%, respectively. The stripe rust resistance gene Yr28 carried on the 4D chromosome was fully expressed in the tetraploid background. The present 4D(4B) disomic substitution line YL-443 was distinguished from the previously reported 4D(4B) lines with the 4D chromosomes from Chinese Spring (CS). Our study demonstrated that YL-443 can be used as elite germplasm for durum wheat breeding targeting high yield potential and stripe rust resistance. The Yr28-specific PCR marker and the 4D chromosome-specific KASP markers together with its unique features of pubescent leaf sheath and auricles can be utilized for assisting selection in breeding.

Karyotype, C-band and NOR phenotype of Anatolian endemic fish Squalius anatolicus (Bogutskaya, 1997) (Teleostei, Leuciscidae)

The karyotype and distribution of constitutive heterochromatin and nucleolus organizer regions (NORs) of Anatolian leuciscine endemic to Lake Beysehir, Squalius anatolicus (Bogutskaya, 1997) were analyzed respectively using conventional Giemsa-staining, C-banding and Ag-impregnation. Diploid chromosome number was 2n = 50 and karyotype consisted of 7 pairs of metacentric, 13 pairs of submetacentric, 5 pairs of subtelo- to acrocentric chromosomes, NF value equaled 90. Heteromorphic elements indicating sex chromosomes were not detected. C-banding revealed clear pericentromeric constitutive heterochromatin blocks in several chromosomes. Ag-impregnation revealed the size heteromorphism of NORs that covered almost the entire short arms of the middle-sized submetacentric chromosome pair. The karyotype pattern and simple NOR phenotype of S. anatolicus are nearly identical with that found not only in Squalius species analyzed to date but also in many other representatives of the Eurasian leuciscine cyprinids, which indicates remarkable chromosome stasis in this leuciscid lineage.

Physical localization of 45S rDNA in Cymbopogon and the analysis of differential distribution of rDNA in homologous chromosomes of Cymbopogon winterianus

Cymbopogon, commonly known as lemon grass, is one of the most important aromatic grasses having therapeutic and medicinal values. FISH signals on somatic chromosome spreads off Cymbopogon species indicated the localization of 45S rDNA on the terminal region of short arms of a chromosome pair. A considerable interspecific variation in the intensity of 45S rDNA hybridization signals was observed in the cultivars of Cymbopogon winterianus and Cymbopogon flexuosus. Furthermore, in all the varieties of Cymbopogon winterianus namely Bio-13, Manjari and Medini, a differential distribution of 45S rDNA was observed in a heterologous pair of chromosome 1. The development of Cymbopogon winterianus var. Manjari through gamma radiation may be responsible for breakage of fragile rDNA site from one of the chromosomes of this heterologous chromosome pair. While, in other two varieties of Cymbopogon winterianus (Bio-13 and Medini), this variability may be because of evolutionary speciation due to natural cross among two species of Cymbopogon which was fixed through clonal propagation. However, in both the situations these changes were fixed by vegetative method of propagation which is general mode of reproduction in the case of Cymbopogon winterianus.

First Report on Nucleolar Organizer Regions (NORs) Polymorphism and Constitutive Heterochromatin of Moonlight Gourami, Trichopodus microlepis (Perciformes, Osphronemidae)

Nucleolar organizer regions (NORs) polymorphism, constitutive heterochromatin and chromosomal analysis of Moonlight gourami, Trichopodus microlepis in Thailand were firstly reported. Specimens were collected from the Chao Phraya and Mekong Basins, Thailand. The mitotic chromosomes were directly prepared from kidney tissues of ten males and ten females. Conventional staining, Ag-NOR banding and C- banding techniques were applied to stain the chromosomes. The results shown that the diploid chromosome number of T. microlepis was 2n=46 and the fundamental number (NF) was 46 in both males and females. The karyotype consisted of 46 telocentric chromosomes classifying as 14 large and 32 medium. No heteromorphic sex chromosome was observed in T. microlepis. The results also exhibited that the interstitial nucleolar organizer regions (NORs) were clearly observed at the long arm of the chromosome pair 7. This is the first report on NORs polymorphism in T. microlepis that a heteromorphic NOR type in one female had a single NOR-bearing chromosome of the chromosome pair 7, whereas 10 males and nine females had two NOR-bearing chromosomes of the chromosome pair 7 with a homomorphic NOR type. Constitutive heterochromatins located at all centromeres of all chromosome pairs. The karyotype formula of T. microlepis is 2n (46) = Lt14 + Mt32.

Meiotic Crossover Patterning

Proper number and placement of meiotic crossovers is vital to chromosome segregation, with failures in normal crossover distribution often resulting in aneuploidy and infertility. Meiotic crossovers are formed via homologous repair of programmed double-strand breaks (DSBs). Although DSBs occur throughout the genome, crossover placement is intricately patterned, as observed first in early genetic studies by Muller and Sturtevant. Three types of patterning events have been identified. Interference, first described by Sturtevant in 1915, is a phenomenon in which crossovers on the same chromosome do not occur near one another. Assurance, initially identified by Owen in 1949, describes the phenomenon in which a minimum of one crossover is formed per chromosome pair. Suppression, first observed by Beadle in 1932, dictates that crossovers do not occur in regions surrounding the centromere and telomeres. The mechanisms behind crossover patterning remain largely unknown, and key players appear to act at all scales, from the DNA level to inter-chromosome interactions. There is also considerable overlap between the known players that drive each patterning phenomenon. In this review we discuss the history of studies of crossover patterning, developments in methods used in the field, and our current understanding of the interplay between patterning phenomena.

Telomere-to-telomere assembly of a fish Y chromosome reveals the origin of a young sex chromosome pair

Background The origin of sex chromosomes requires the establishment of recombination suppression between the proto-sex chromosomes. In many fish species, the sex chromosome pair is homomorphic with a recent origin, providing species for studying how and why recombination suppression evolved in the initial stages of sex chromosome differentiation, but this requires accurate sequence assembly of the X and Y (or Z and W) chromosomes, which may be difficult if they are recently diverged. Results Here we produce a haplotype-resolved genome assembly of zig-zag eel (Mastacembelus armatus), an aquaculture fish, at the chromosomal scale. The diploid assembly is nearly gap-free, and in most chromosomes, we resolve the centromeric and subtelomeric heterochromatic sequences. In particular, the Y chromosome, including its highly repetitive short arm, has zero gaps. Using resequencing data, we identify a ~7 Mb fully sex-linked region (SLR), spanning the sex chromosome centromere and almost entirely embedded in the pericentromeric heterochromatin. The SLRs on the X and Y chromosomes are almost identical in sequence and gene content, but both are repetitive and heterochromatic, consistent with zero or low recombination. We further identify an HMG-domain containing gene HMGN6 in the SLR as a candidate sex-determining gene that is expressed at the onset of testis development. Conclusions Our study supports the idea that preexisting regions of low recombination, such as pericentromeric regions, can give rise to SLR in the absence of structural variations between the proto-sex chromosomes.

Blocking Protein Phosphatase 1 [PP1] Prevents Loss of Tether Elasticity in Anaphase Crane-Fly Spermatocytes

In normal anaphase cells, telomeres of each separating chromosome pair are connected to each other by tethers. Tethers are elastic at the start of anaphase: arm fragments cut from anaphase chromosomes in early anaphase move across the equator to the oppositely-moving chromosome, telomere moving toward telomere. Tethers become inelastic later in anaphase as the tethers become longer: arm fragments no longer move to their partners. When early anaphase cells are treated with Calyculin A (CalA), an inhibitor of protein phosphatases 1 (PP1) and 2A (PP2A), at the end of anaphase chromosomes move backward from the poles, with telomeres moving toward partner telomeres. Experiments described herein show that in cells treated with CalA, backwards movements are stopped in a variety of ways, by cutting the tethers of backwards moving chromosomes, by severing arms of backwards moving chromosomes, by severing arms before the chromosomes reach the poles, and by cutting the telomere toward which a chromosome is moving backwards. Measurements of arm-fragment velocities show that CalA prevents tethers from becoming inelastic as they lengthen. Since treatment with CalA causes tethers to remain elastic throughout anaphase and since inhibitors of PP2A do not cause the backwards movements, PP1 activity during anaphase causes the tethers to become inelastic.