Genetic Features of Triticale–Wheat Hybrids with Vaviloid-Type Spike Branching

Vaviloid spike branching, also called sham ramification, is a typical trait of Triticum vavilovii Jakubz. and is characterized by a lengthening of the spikelet axis. In this article, we present the results of a study of three triticale–wheat hybrid lines with differences in terms of the manifestation of the vaviloid spike branching. Lines were obtained by crossing triticale with hexaploid wheat, T. aestivum var. velutinum. The parental triticale is a hybrid of synthetic wheat (T. durum × Ae. tauschii var. meyrei) with rye, S. cereale ssp. segetale. Line 857 has a karyotype corresponding to hexaploid wheat and has a spike morphology closest to normal, whereas Lines 808/1 and 844/4 are characterized by the greatest manifestation of vaviloid spike branching. In Lines 808/1 and 844/4, we found the substitution 2RL(2DL). The karyotypes of the latter lines differ in that a pair of telocentric chromosomes 2DS is detected in Line 808/1, and these telocentrics are fused into one unpaired chromosome in Line 844/4. Using molecular genetic analysis, we found a deletion of the wheat domestication gene Q located on 5AL in the three studied hybrid lines. The deletion is local since an analysis of the adjacent gene B1 showed the presence of this gene. We assume that the manifestation of vaviloid spike branching in two lines (808/1 and 844/4) is associated with a disturbance in the joint action of genes Q and AP2L2-2D, which is another important gene that determines spike morphology and is located on 2DL.

TRF1 Depletion Reveals Mutual Regulation Between Telomeres, Kinetochores, and Inner Centromeres in Mouse Oocytes

In eukaryotic chromosomes, the centromere and telomere are two specialized structures that are essential for chromosome stability and segregation. Although centromeres and telomeres often are located in close proximity to form telocentric chromosomes in mice, it remained unclear whether these two structures influence each other. Here we show that TRF1 is required for inner centromere and kinetochore assembly in addition to its role in telomere protection in mouse oocytes. TRF1 depletion caused premature chromosome segregation by abrogating the spindle assembly checkpoint (SAC) and impairing kinetochore-microtubule (kMT) attachment, which increased the incidence of aneuploidy. Notably, TRF1 depletion disturbed the localization of Survivin and Ndc80/Hec1 at inner centromeres and kinetochores, respectively. Moreover, SMC3 and SMC4 levels significantly decreased after TRF1 depletion, suggesting that TRF1 is involved in chromosome cohesion and condensation. Importantly, inhibition of inner centromere or kinetochore function led to a significant decrease in TRF1 level and telomere shortening. Therefore, our results suggest that telomere integrity is required to preserve inner centromere and kinetochore architectures, and vice versa, suggesting mutual regulation between telomeres and centromeres.

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.

MORPHOLOGICAL AND CHROMOSOMAL CHARACTERIZATION OF ORCHID Peristylus goodyeroides Lindl. FROM CURUG SETAWING, KULONPROGO

Peristylus goodyeroides is a terestrial orchid that scattered around Southeast Asia. Morphological characters of P. goodyeroides can vary, depending on the ecological factors and habitat. Cytological characters in the form of chromosome configurations can be used as a taxonomic tool for the process of identifying and understanding variations in taxa. The purpose of this study was to determine the morphological characters and chromosome configuration of the P. goodyeroides from Curug Setawing, Kulonprogo. The method used was morphological characterization and plant chromosome preparation by squash method with the main steps of fixation, maceration, staining and observation. Data were analyzed with the help of Image raster 3, Corel Draw X7, and Microsoft Excel 2013. P. goodyeroides from Curug Setawing has the morphological characters of root tubers, cylindrical stems, ovate leaf shape, convolutive leaf arrangement and creamy white flowers. The orchid has a number of chromosomes 2n = 10 with a karyotype formula of 2n = 2x = 8m + 2t. Metacentric chromosomes are found on chromosomes 1-8 and telocentric chromosomes 9-10. The absolute arm length of the chromosomes has a range of 2.03-3.44 μm, the relative arm length of the chromosomes is 2.21-3.32 μm, the length of the p arm is 1.13-1.58 μm and the q arm is 1.23-2.12 μm.

First cytogenetic characterization of the Amazon Catfish Leiarius marmoratus (Gill, 1870) and its hybrid with Pseudoplatystoma reticulatum (Eigenmann & Eigenmann, 1889)

This study reports the first cytogenetic characterization of the Amazonian catfish Leiarius marmoratus (“jandiá”) and its F1 (first generation) hybrid “cachandiá” with Pseudoplatystoma reticulatum (“cachara”). A diploid number of 56 chromosomes and a single argyrophilic nucleolus organizer region (Ag-NOR) in the short arm of two sub-telocentric chromosomes were observed for both L. marmoratus and P. reticulatum, but with differences in the karyotype formula and the size of the chromosome pair with NORs. The hybrid showed 2n = 56 chromosomes with an intermediate karyotype when compared to the parental species. A single Ag-NOR was maintained in the hybrid but located in two chromosomes with marked differences in size and presenting intraindividual variation in NOR activity (nucleolar dominance). For L. marmoratus and the hybrid, heterochromatic bands were predominately distributed in the terminal, centromeric, and sub-centromeric regions of some chromosomes and 5S rDNA sites located in two distinct sub-telocentric chromosomes, similar to the previously described for P. reticulatum. The data suggested that the hybrid karyotype might be insufficient for a precise discrimination of hybrids, however, Ag-NOR can be used as a chromosome marker to differentiate “cachandiá” from L. marmoratus and P. reticulatum. The current study also provides insights into the chromosomal features of L. marmoratus and contributes with novel cytogenetic information of this native Amazonian catfish included in the Pimelodidae family.

First report of karyological analysis and heteromorphic nucleolar organizer region of Black Surgeonfish (Acanthurus gahhm, Acanthuridae) in Thailand

This research was the first report on karyological analysis and heteromorphic nucleolar organizer region of black surgeonfish (Acanthurus gahhm, Acanthuridae) in Thailand. The 10 male and 10 female specimens were collected from Phuket Marine Biological Center, and Phang Nga Coastal Research and Development Center, Andaman Sea, Thailand. Mitotic chromosomes were directly prepared from gill and kidney tissues. The chromosomes were stained by conventional Giemsa staining and Ag-NOR banding techniques. Results showed that the diploid chromosomes number of A. gahhm was 2n=48, the fundamental numbers (NF) was 54 in both male and female. The karyotype consist of 6 large acrocentric, 20 large telocentric, 18 medium telocentric and 4 small telocentric chromosomes. None of strange size chromosomes related to sex was found. The heteromorphic nucleolar organizer regions (NORs) were observed on telomeric short arm of first acrocentric which can defined as 1a1b. There is NOR in 1a and not in 1b. The karyotype formula of black surgeon fish was as follows: 2n (48) = La6+Lt20+Mt18+St4

Comparative cytogenetic analysis of fishes in the genus Trichopodus (Osphronemidae) in Thailand

Supiwong W, Wongchantra P, Thongnetr W, Mingkwan B, Chaiyasan P, Pinmongkhonkul S, Pinthong K, Tanomtong A. 2021. Comparative cytogenetic analysis of fishes in the genus Trichopodus (Osphronemidae) in Thailand. Biodiversitas 22: 3029-3036. Comparative cytogenetic study of four species of the genus Trichopodus including T. leerii, T. microlepis, T. pectoralis, and T. trichopterus from Thailand, was carried out. The specimens were collected from the Basins throughout Thailand. Chromosome preparation was directly performed from the kidney tissues. Conventional staining by Giemsa solution, Ag–NOR banding by silver nitrate solution and C-banding by NaOH solution were conducted to stain the chromosomes. Results showed that four species studied have the same diploid number and fundamental number as 46 composing of all telocentric chromosomes. Karyotypes consisting of large-medium-small sizes in T. leerii, T. microlepis, T. pectoralis and T. trichopterus were 16–28–2, 14–32–0, 20–26–0 and 10–30–6 chromosomes, respectively. The marker chromosomes which present the NOR positions are the single pair in all species but there are differences in the locations and pairs such as pairs no. 1, 7, 2 and 1, respectively. Most species except T. trichopterus, NOR sites locate at interstitial region adjacent to the centromere. T. trichopterus had the telomeric NORs. Constitutive heterochromatin blocks displayed at the centromeric/pericentromeric regions of all chromosomes in T. leerii and T. microlepis whereas in T. pectoralis and T. trichopterus, those presented at not only centromeres of several chromosome pairs, but they also were found at interstitial sites. The obtained finding, cytogenetic data has species-specific. Thus, it can be used for further study on taxonomy. Moreover, this data shows the close relationship of genetics among these species so that the application for breeding may be used in the future.

Establishment of Cell Lines from Slender Walking Catfish Clarias nieuhofii (Valenciennes, 1840)

The slender walking catfish (Clarias nieuhofii Valenciennes, 1840) is a freshwater fish found in the ecosystems within the peat swamp forests of South-East Asian countries, including Thailand. In this study, a new testis fibroblast-like (FLT) and ovary epithelial-like (ELO) cells derived from C. nieuhofii (Valenciennes, 1840) was established. The optimal number of both cells was determined, and the optimal starting concentration of both cells is approximately 105 cellsžml-1. The FLT and ELO can grow best at 28 °C in DMEM supplemented with 20, 15, and 10 % fetal bovine serum (FBS). Chromosome analysis revealed that the typical diploid chromosome number was 66 (2n) in C. nieuhofii (Valenciennes, 1840). In both males and females, the elementary number of chromosomes (NF; the number of chromosome arms) was 98. No heteromorphic chromosome was observed. Four types of chromosomes were found in this species, including 14 metacentric, 20 submetacentric, 2 acrocentric, and 30 telocentric chromosomes. The FLT and ELO cells were maintained up to 6 months with more than 20 passages successfully cryopreserved and thawed. HIGHLIGHTS Two new fibroblast-like cell from testis and epithelium-like cell from ovary, derived from slender walking catfish Clarias nieuhofii (Valenciennes, 1840) have been successfully established by explant outgrowth technique Indicated the optimal cell density, fetal bovine serum and types of culture media for growing of both cells Chromosome analysis revealed that the typical diploid chromosome number is 66 (2n) in Clarias nieuhofii (Valenciennes, 1840) GRAPHICAL ABSTRACT

FISH Identifies Chromosome Differentiation Between Contemporary Genomes of Wild Types and the Ancestral Genome of Unisexual Clones of Dojo Loach, Misgurnus anguillicaudatus

In dojo loach (<i>Misgurnus anguillicaudatus</i>), although most wild types are gonochoristic diploids that are genetically differentiated into 2 groups, A and B, clonal lineages appear in certain localities. Clonal loaches have been considered to have hybrid origins between the 2 groups by a series of genetic studies. In this study, using FISH with a newly developed probe (ManDra-A), we identified 26 (1 pair of metacentric and 12 pairs of telocentric chromosomes) of 50 diploid chromosomes in contemporary wild-type group A loach. In contrast, ManDra-A signals were not detected on metacentric chromosomes derived from the ancestral group A of clonal loach. The FISH results clearly showed the presence of certain differentiations in metacentric chromosomes between ancestral and contemporary group A loach. Two-color FISH with ManDra-A and group B-specific ManDra (renamed ManDra-B) probes reconfirmed the hybrid origin of clones by identifying chromosomes from both groups A and B in metaphases. Our results showed the hybrid origin of clonally reproducing fish and the possibility that chromosomal differentiation between ancestral and contemporary fish can affect gametogenesis. In meiotic spermatocytes of sex-reversed clones, ManDra-A, and not ManDra-B, signals were detected in 12 out of 50 bivalents. Thus, the results further support the previous conclusion that clonal gametogenesis was assured by pairing between sister chromosomes duplicated from each ancestral chromosome from group A or B. Our study deepens the knowledge about the association between clonality and hybridity in unisexual vertebrates.

Pericentromeric heterochromatin impacts genome compartmentalization and sex chromosome evolution in a fish

Compartmentalization is one of the principles of chromosome 3D organization and has been suggested to be driven by the attraction of heterochromatin. The extent to which the pericentromeric heterochromatin (PCH) impacts chromosome compartmentalization is yet unclear. Here we produced a chromosome-level and fully phased diploid genome of an aquaculture fish, zig-zag eel (Mastacembelus armatus), and identified the centromeric and pericentromeric regions in the majority of chromosomes of both haploid genomes. The PCH is on average 4.2 Mb long, covering 17.7% of the chromosomes, and is the major target of histone 3 lysine 9 trimethylation (H3K9me3). In nearly half of the chromosomes, the PCH drives the chromosomes into two or three megascale chromatin domains with the PCH being a single one. We further demonstrate that PCH has a major impact in submetacentric, metacentric and small telocentric chromosomes in which the PCH drives the distribution of active and inactive compartments along the chromosomes. Additionally, we identified the young and homomorphic XY sex chromosomes that are submetacentric with the entire short-arm heterochromatinized. Interestingly, the sex-determining region seems to arise within the PCH that has been in place prior to the X-Y divergence and recombination suppression. Together, we demonstrate that the PCH can cover a considerably large portion of the chromosomes, and when it does so, it drives chromosome compartmentalization; and we propose a new model for the origin and evolution of homomorphic sex chromosomes in fish.