Karyotype 2n Research Articles

Development of Apomictic 56-Chromosomal Maize-Tripsacum Hybrids: A Potential Breakthrough in Heterosis Fixation.

Maize (Zea mays L.) is one of the most demanded grain crops in the world. Currently, production has exceeded one billion tons and is increasing by 3-5% annually. Such growth is due to the genetic potential of the crop and the use of heterosis F1 hybrids in production. However, the need to produce first-generation seed annually poses significant challenges and is an economically costly technology. A solution to this problem may be the transfer of the asexual (apomictic) mode of reproduction to maize from its wild relative, eastern gamagrass (Tripsacum dactyloides L.). In this work, we report the production of 56-chromosome apomictic hybrids of maize (Zea mays L.) with eastern gamagrass (T. dactyloides L.) with restored anther fertility. The mode of reproduction of the plant was confirmed by counting chromosomes and sequencing the nuclear gene (Pox3) and chloroplast tRNA-Leu (trnL) gene. These apomictic hybrids had karyotypes of 2n = 56 = [(10Zm(573MB) + 36Td) + 10Zm(611CB)] and 2n = 56 = [(10Zm(611CB) + 36Td) + 10Zm(611CB)]. The resulting hybrids can be widely used as a fodder crop.

Long-term preservation of established fibroblast lines from six-banded armadillos (Euphractus sexcintus, Linnaeus, 1758) by extended passage and cryopreservation.

Establishing new somatic cell cultures has raised significant attention as an effective and convenient way to preserve genetic samples for different applications. Although many lines have been established in model animals, none derived from six-banded armadillo species is currently available. We report the successful isolation and characterization of fibroblasts from six-banded armadillos, evaluating the cell quality after extended culture and cryopreservation. Initially, we collected ear skin from five captive adult individuals and identified fibroblast lines by morphology, karyotyping, and immunophenotyping assays. The isolated fibroblasts were evaluated after several passages (fourth, seventh, and tenth passages) and cryopreservation by slow freezing. Cell morphology, viability, metabolism, proliferative activity, mitochondrial membrane potential, and apoptosis levels were analyzed. The skin explants had great adhesion, and cell outgrowth could be seen after 3-6 d. The cells were verified as fibroblasts at the fourth passage by vimentin expression and normal karyotype (2n = 58). The viability remained high (> 87%) and constant from the fourth to the tenth passage (p > 0.05). The passages did not change the cell morphology and metabolic and growth rates. Moreover, cryopreservation did not affect most evaluated parameters; post-thawed cells maintained their viability, growth, metabolism, and apoptosis levels. Nevertheless, cryopreservation increased mitochondrial membrane permeability and cell population doubling time compared to non-cryopreserved cells (p < 0.05). In summary, viable fibroblasts can be obtained from six-banded armadillo skin while conserving their quality as the number of passages increases and featuring few changes after cryopreservation.

Integrating Genomic and Chromosomal Data: A Cytogenetic Study of Transancistrus santarosensis (Loricariidae: Hypostominae) with Characterization of a ZZ/ZW Sex Chromosome System.

The plecos (Loricariidae) fish represent a great model for cytogenetic investigations due to their variety of karyotypes, including diploid and polyploid genomes, and different types of sex chromosomes. In this study we investigate Transancistrus santarosensis a rare loricariid endemic to Ecuador, integrating cytogenetic methods with specimens' molecular identification by mtDNA, to describe the the species karyotype. We aim to verify whether sex chromosomes are cytologically identifiable and if they are associated with the accumulation of repetitive sequences present in other species of the family. The analysis of the karyotype (2n = 54 chromosomes) excludes recent centric fusion and pericentromeric inversion and suggests the presence of a ZZ/ZW sex chromosome system at an early stage of differentiation: the W chromosome is degenerated but is not characterized by the presence of differential sex-specific repetitive DNAs. Data indicate that although T. santarosensis has retained the ancestral diploid number of Loricariidae, it accumulated heterochromatin and shows non-syntenic ribosomal genes localization, chromosomal traits considered apomorphic in the family.

Microchromosome fusions underpin convergent evolution of chameleon karyotypes.

Evolutionary shifts in chromosome compositions (karyotypes) are major drivers of lineage and genomic diversification. Fusion of ancestral chromosomes is one hypothesized mechanism for the evolutionary reduction of the total chromosome number, a frequently implied karyotypic shift. Empirical tests of this hypothesis require model systems with variable karyotypes, known chromosome features, and a robust phylogeny. Here we used chameleons, diverse lizards with exceptionally variable karyotypes ($2n=20\text{-}62$), to test whether chromosomal fusions explain the repeated evolution of karyotypes with fewer chromosomes than ancestral karyotypes. Using a multidisciplinary approach including cytogenetic analyses and phylogenetic comparative methods, we found that a model of constant loss through time best explained chromosome evolution across the chameleon phylogeny. Next, we tested whether fusions of microchromosomes into macrochromosomes explained these evolutionary losses using generalized linear models. Multiple comparisons supported microchromosome fusions as the predominant agent of evolutionary loss. We further compared our results to various natural history traits and found no correlations. As such, we infer that the tendency of microchromosomes to fuse was a quality of the ancestral chameleon genome and that the genomic predisposition of ancestors is a more substantive predictor of chromosome change than the ecological, physiological, and biogeographical factors involved in their diversification.

Exploration of Parascaris species in three different Equus populations in China

BackgroundThe roundworms, Parascaris spp., are important nematode parasites of foals and were historically model organisms in the field of cell biology, leading to many important discoveries. According to karyotype, ascarids in Equus are commonly divided into Parascaris univalens (2n = 2) and Parascaris equorum (2n = 4).MethodsHere, we performed morphological identification, karyotyping and sequencing of roundworms from three different hosts (horses, zebras and donkeys). Phylogenetic analysis was performed to study the divergence of these ascarids based on cytochrome c oxidase subunit I (COI) and internal transcribed spacer (ITS) sequences.ResultsKaryotyping, performed on eggs recovered from worms of three different Equus hosts in China, showed two different karyotypes (2n = 2 in P. univalens collected from horses and zebras; 2n = 6 in Parascaris sp. collected from donkeys). There are some differences in the terminal part of the spicula between P. univalens (concave) and Parascaris sp. (rounded). Additionally, it was found that the egg’s chitinous layer was significantly thicker in Parascaris sp. (> 5 μm) than P. univalens (< 5 μm) (F(2537) = 1967, P < 0.01). Phylogenetic trees showed that the sequences of Parascaris from Equus hosts were divided into two distinct lineages based on sequences of the COI and ITS.ConclusionsComparing the differences in roundworms collected from three different Equus hosts, this study describes a Parascaris species (Parascaris sp.) with six chromosomes in donkeys. It is worth noting that the thickness of the chitinous layer in the Parascaris egg may serve as a diagnostic indicator to distinguish the two roundworms (P. univalens and Parascaris sp.). The Parascaris sp. with six chromosomes in donkeys in the present study may be a species of P. trivalens described in 1934, but the possibility that it is a new Parascaris species cannot be ruled out. Both karyotyping and molecular analysis are necessary to solve the taxonomic problems in Parascaris species.

Effect of colchicine mutagen on phenotype and genotype of Vigna unguiculata var. sesquipedalis the 7th generation

Abstract. Fathurrahman F, Mardaleni, Krisianto A. 2023. Title Effect of colchicine mutagen on phenotype and genotype of Vigna unguiculata var. Sesquipedalis the 7th generation. Biodiversitas 24: 1408-1416. The problem with long bean species is the low diversity of varieties, so obtaining them using chromosomal mutations was necessary. One of the ingredients used was colchicine. The aim of this study was to determine the effectiveness of the mutagen colchicine against mutations that occur in Vigna unguiculata var. sesquipedalis the 7th generation (renek long bean). The design used was a factorial Completely Randomized Design consisting of two factors. The first factor was four colchicine concentration levels: including without colchicine (the control) and colchicine concentration (0.01%, 0.05%, and 0.09%). The second factor was the soaking of the V. unguiculata var. sesquipedalis seeds, namely without soaking of seeds and soaking seeds (6, 12, 18, and 24 hours). Further phenotype analysis was tested statistically using the DMRT test at p &lt; 0.05, while genotypes were analyzed using karyotypes and statistical T-test at p &lt; 0.05. The results showed that treatment with 0.09% of colchicine and soaking for 24 hours was the best combination. Phenotype analysis showed increased growth with colchicine treatment: plant height increased by 20.59%, pod weight increased by 0.88%, the number of pods increased by 47.93%, pod length increased by 22.50%, and 100 seed weight increased by 15.91%. While Genotype analysis obtained a total of 11 karyotype chromosomes and 2n = 22 chromosomes. Chromosome karyotypes obtained two types, namely metacentric and telocentric. The arm length (p+q) in the 0.09% colchicine treatment sample and 24-hour soaking time was 1.77 µm longer than the control sample, which was 1.53 µm. It turns out that colchicine can increase the growth and changes in the structure of chromosomes so that a mutated V. unguiculata var. sesquipedalis species is obtained.

Whole-Genome Analysis Reveals the Dynamic Evolution of Holocentric Chromosomes in Satyrine Butterflies.

Butterfly chromosomes are holocentric, i.e., lacking a localized centromere. Potentially, this can lead to rapid karyotypic evolution through chromosome fissions and fusions, since fragmented chromosomes retain kinetic activity, while fused chromosomes are not dicentric. However, the actual mechanisms of butterfly genome evolution are poorly understood. Here, we analyzed chromosome-scale genome assemblies to identify structural rearrangements between karyotypes of satyrine butterfly species. For the species pair Erebia ligea-Maniola jurtina, sharing the ancestral diploid karyotype 2n = 56 + ZW, we demonstrate a high level of chromosomal macrosynteny and nine inversions separating these species. We show that the formation of a karyotype with a low number of chromosomes (2n = 36 + ZW) in Erebia aethiops was based on ten fusions, including one autosome-sex chromosome fusion, resulting in a neo-Z chromosome. We also detected inversions on the Z sex chromosome that were differentially fixed between the species. We conclude that chromosomal evolution is dynamic in the satyrines, even in the lineage that preserves the ancestral chromosome number. We hypothesize that the exceptional role of Z chromosomes in speciation may be further enhanced by inversions and sex chromosome-autosome fusions. We argue that not only fusions/fissions but also inversions are drivers of the holocentromere-mediated mode of chromosomal speciation.

Ranunculusmaoxianensis (Ranunculaceae), a new species from northwestern Sichuan, China, with an emended description of R.chongzhouensis, the putative closest ally of the new species.

Ranunculusmaoxianensis (Ranunculaceae), a new species from Jiuding Shan in Maoxian county, northwestern Sichuan province, China, is here illustrated and described. The species is morphologically similar to R.chongzhouensis, a species also occurring in Sichuan, in having reniform leaves and puberulous receptacles, carpels and achenes, but differs by having leaves adaxially puberulous with shorter appressed hairs 0.16‒0.28 mm long (vs. longer appressed hairs 0.55‒0.85 mm long), larger flowers (1.8‒2 cm vs. 1.4‒1.6 cm in diameter), larger (8‒10 × 5.5‒6.5 mm vs. 6‒7 × 4.5‒5 mm) and widely obovate petals (vs. obovate), more numerous stamens (35‒55 vs. 12‒18), and subglobose gynoecium and aggregate fruit (vs. ellipsoid). The two species are also different in chromosome number and chromosome morphology. Ranunculuschongzhouensis has a karyotype of 2n = 2x = 16 = 10m + 6sm while R.maoxianensis has a karyotype of 2n = 4x = 32 = 16m + 16sm. An emended description of R.chongzhouensis is provided, and its geographical distribution is largely extended.

Molecular Cytogenetic Characterization of C-Band-Positive Heterochromatin of the Greater Long-Tailed Hamster (Tscherskia triton, Cricetinae)

The greater long-tailed hamster (Tscherskia triton, Cricetinae) has a unique karyotype (2n = 28), containing 11 pairs of acrocentric chromosomes with large C-band-positive centromeric heterochromatin blocks. To understand the origin and evolutionary process of heterochromatin in this species, we isolated novel families of chromosome site-specific highly repetitive DNA sequences from TaqI-digested genomic DNA and then characterized them by chromosome in situ and filter hybridization. The TaqI-families of repetitive sequences were classified into 2 types by their genome organization and chromosomal distribution: the 110-bp repeated sequence organized in large tandem arrays (as satellite DNA), localized to centromeric C-positive heterochromatin of acrocentric autosomes (chromosomes 1–11) and submetacentric X chromosome, and the 405-bp repeated sequence that was composed of 30–32-bp internal repeats, distributed in the pericentromeric region on the short arms of X and Y chromosomes. The repetitive sequences did not cross-hybridize with genomic DNA of any genera of Cricetinae (Mesocricetus, Cricetulus, and Phodopus). These results suggest that the 110-bp and 405-bp repeats rapidly diverged in the lineage of T. triton, evolving in a concerted manner among autosomes and X chromosome and within X and Y chromosomes, respectively. The 110-bp centromeric repeat contained a 17-bp motif in which 9 bases are essential for binding with the centromere-associated protein CENP-B, suggesting the possibility that the 110-bp major satellite DNA carrying the 17-bp motif may have a role in the formation of specified structure and/or function of centromeres in T. triton.

Identification of Iguania Ancestral Syntenic Blocks and Putative Sex Chromosomes in the Veiled Chameleon (Chamaeleo calyptratus, Chamaeleonidae, Iguania)

The veiled chameleon (Chamaeleo calyptratus) is a typical member of the family Chamaeleonidae and a promising object for comparative cytogenetics and genomics. The karyotype of C. calyptratus differs from the putative ancestral chameleon karyotype (2n = 36) due to a smaller chromosome number (2n = 24) resulting from multiple chromosome fusions. The homomorphic sex chromosomes of an XX/XY system were described recently using male-specific RADseq markers. However, the chromosomal pair carrying these markers was not identified. Here we obtained chromosome-specific DNA libraries of C. calyptratus by chromosome flow sorting that were assigned by FISH and sequenced. Sequence comparison with three squamate reptiles reference genomes revealed the ancestral syntenic regions in the C. calyptratus chromosomes. We demonstrated that reducing the chromosome number in the C. calyptratus karyotype occurred through two fusions between microchromosomes and four fusions between micro-and macrochromosomes. PCR-assisted mapping of a previously described Y-specific marker indicates that chromosome 5 may be the sex chromosome pair. One of the chromosome 5 conserved synteny blocks shares homology with the ancestral pleurodont X chromosome, assuming parallelism in the evolution of sex chromosomes from two basal Iguania clades (pleurodonts and acrodonts). The comparative chromosome map produced here can serve as the foundation for future genome assembly of chameleons and vertebrate-wide comparative genomic studies.

First Cytogenetic Study Through Conventional Staining of The Ant Genus Blepharidatta Wheeler, 1915 (Hymenoptera: Formicidae: Attini)

Blepharidatta is a rare Neotropical ant genus in the Attini tribe of the subfamily Myrmicinae. It has only four valid species and among them Blepharidatta delabiei was recently described and there is little knowledge on its biology. This study is the first cytogenetic characterization for the genus Blepharidatta and also presents the biology of B. delabiei. Cytogenetic analyses revealed a karyotype 2n = 28 with acrocentric and metacentric chromosomes and a karyotypic formula (m: metacentric, a: acrocentric): 2K = 16m + 12a. We observed that ants of this species have diurnal habits with higher foraging activity in the afternoon and are possibly omnivorous as they accepted the baits used. The distance between colonies varied from 5 to 7 meters.

A new species of Akodon Meyen, 1833 (Rodentia: Cricetidae) from dry forests of the Amazonia-Cerrado transition

Akodon is the most diverse genus of the tribe Akodontini, comprising 41 extant species. Although distributed in a wide range of environments in South America, the genus was long considered to be absent from Amazon Forest. Nonetheless, in recent decades, records of Akodon have been reported based on specimens from southern Amazonia, throughout the contact zone with Cerrado. Published data about the unique karyotype (2n=10), along with evidence of sister phylogenetic relationship (Cytb sequences) with Akodon cursor, a species from the Atlantic Forest, suggest that these specimens represent an undescribed species. Herein we hypothesize that these samples represent a new species of Akodon from Amazonia-Cerrado transitional areas. Through the analyses of 217 specimens from Mato Grosso and Pará states, Brazil, as well as 336 specimens of Akodon cursor species group from the Atlantic Forest, we provide qualitative and quantitative external and craniodental data that support their distinction in comparison with other congeners. The sister relationship between this new species and A. cursor provide additional evidence of a past connection between the two largest tropical forests of South America. Moreover, the recognition of Akodon n. sp. points to the presence of the genus in the Amazonia-Cerrado transition, an area currently under great anthropic pressure, highlighting the fact that we might be losing a portion of Brazilian biodiversity before even getting to know it.

The first high-density genetic map of common cockle (Cerastoderma edule) reveals a major QTL controlling shell color variation

Shell color shows broad variation within mollusc species and despite information on the genetic pathways involved in shell construction and color has recently increased, more studies are needed to understand its genetic architecture. The common cockle (Cerastoderma edule) is a valuable species from ecological and commercial perspectives which shows important variation in shell color across Northeast Atlantic. In this study, we constructed a high-density genetic map, as a tool for screening common cockle genome, which was applied to ascertain the genetic basis of color variation in the species. The consensus genetic map comprised 19 linkage groups (LGs) in accordance with the cockle karyotype (2n = 38) and spanned 1073 cM, including 730 markers per LG and an inter-marker distance of 0.13 cM. Five full-sib families showing segregation for several color-associated traits were used for a genome-wide association study and a major QTL on chromosome 13 associated to different color-traits was detected. Mining on this genomic region revealed several candidate genes related to shell construction and color. A genomic region previously reported associated with divergent selection in cockle distribution overlapped with this QTL suggesting its putative role on adaptation.

Karyotype of the Mormyrid Baby Whale Fish (Osteoglossiformes: Mormyridae): Brienomyrus Brachyistius in Oluwa River, Nigeria

The mormyrid fishes of the order Osteoglossiformes are abundant in the freshwaters of tropical Africa; the enormous species diversity of the group requires many systematics tools to comprehend this diversity better. Cytogenetics data can complement other systematics procedures to accomplish the above-stated objective, but information on many mormyrids' karyotypes is unavailable. In this study, Brienomyrus brachyistius from the Oluwa River was studied to enhance the understanding of the karyotype organization in the Mormyridae. Metaphase chromosomes were processed from the fish kidney following a conventional cytogenetic procedure; the fish was subjected to an intra-peritoneal colchicine injection at the rate of 1ml per 100g of fish mass. Hypotonic treatment was in 0.56% KCl solution, and cells fixation and washes were in 3: 1 methanol: acetic acid. The slides were stained with Giemsa solution, while the constitutive heterochromatin was revealed from the previous Giemsa stained slide using the C-banding sequential method. Capturing of metaphase images was done with an Olympus BX50 microscope. The karyotype of 2n = 50 (50sta) obtained in the species was the first mormyrid karyotype composed of 2n = 50 acrocentrics, supporting the proposed ancestral karyotype for the Mormyridae. Keywords: Cytogenetics, chromosomes, Osteoglossiformes, karyotype, Mormyrids, elephantfish DOI: 10.7176/JBAH/12-20-04 Publication date: October 31 st 2022

A new species of Cephalophlugis Gorochov, 1998 (Orthoptera: Tettigoniidae: Meconematinae: Phlugidini) from Southern Brazil, with bioacoustics and cytogenetics.

Cephalophlugis Gorochov, 1998 is a Neotropical genus of predatory katydids with two valid species, Cephalophlugis cephalotes (Bolvar, 1888) (from So Paulo, Brazil) and Cephalophlugis setosa Gorochov, 2012 (from Sucumbios, Ecuador). Herein, we describe a third species of the genus, Cephalophlugis gaucho sp. nov., from Rio Grande do Sul, Brazil. This species is easily distinguished from the others by the color of the hind wings and the number of spurs on the fore and middle legs. The calling song is composed of a long phrase with a series of syllables that vary in emission rate from beginning to end, with a dominant frequency of 23.43 0.6 (22.1224.37, n=11) kHz. So far, the species sings at the lowest frequency among the Phlugidini. From the chromosomal perspective, C. gaucho sp. nov. presents the karyotype 2n=31, X0, with the complement conserved within Meconematinae.

Genomic analyses of wild argali, domestic sheep, and their hybrids provide insights into chromosome evolution, phenotypic variation, and germplasm innovation.

Understanding the genetic mechanisms of phenotypic variation in hybrids between domestic animals and their wild relatives may aid germplasm innovation. Here, we report the high-quality genome assemblies of a male Pamir argali (O. ammon polii, 2n = 56), a female Tibetan sheep (O. aries, 2n = 54), and a male hybrid of Pamir argali and domestic sheep, and the high-throughput sequencing of 425 ovine animals, including the hybrids of argali and domestic sheep. We detected genomic synteny between Chromosome 2 of sheep and two acrocentric chromosomes of argali. We revealed consistent satellite repeats around the chromosome breakpoints, which could have resulted in chromosome fusion. We observed many more hybrids with karyotype 2n = 54 than with 2n = 55, which could be explained by the selfish centromeres, the possible decreased rate of normal/balanced sperm, and the increased incidence of early pregnancy loss in the aneuploid ewes or rams. We identified genes and variants associated with important morphological and production traits (e.g., body weight, cannon circumference, hip height, and tail length) that show significant variations. We revealed a strong selective signature at the mutation (c.334C > A, p.G112W) in TBXT and confirmed its association with tail length among sheep populations of wide geographic and genetic origins. We produced an intercross population of 110 F2 offspring with varied number of vertebrae and validated the causal mutation by whole-genome association analysis. We verified its function using CRISPR-Cas9 genome editing. Our results provide insights into chromosomal speciation and phenotypic evolution and a foundation of genetic variants for the breeding of sheep and other animals.

Asterquanzhouensis (Asteraceae), a new riparian species from eastern China.

Asterquanzhouensis sp. nov. (Asteraceae) from Fujian, eastern China, is described and illustrated. It grows on rocks in the riparian zone. Morphological, cytological and molecular investigations of A.quanzhouensis were carried out. The morphological data and phylogenetic analysis based on combined ITS, ETS and trnL-F dataset suggest that A.quanzhouensis is a separate species closely related to A.tonglingensis. The new species differs from the latter by the shorter stem length, leaf morphology, colour of phyllaries, number of ray florets, and achene shape. The cytological observation shows that the new species is diploid with a karyotype of 2n = 18.

Description of a model tardigrade Paramacrobiotus metropolitanus sp. nov. (Eutardigrada) from Japan with a summary of its life history, reproduction and genomics

The genus Paramacrobiotus was erected in 2009 from the genus Macrobiotus, and 43 Paramacrobiotus species have been described to date. Although the first genome sequence in the genus was reported for the TYO strain of Paramacrobiotus sp., which is a dioecious species and has five bivalent chromosomes, its precise taxonomic identification remained undetermined. Here, we report its morphology, confirming the presence of a microplacoid, cuticular bulge on the inner side of legs IIII, and granulation on the inner side of legs IV under both light and electron microscopy, and smooth areoles on the egg shell, indicating that it differs from other described species. In addition, the previously described karyotype 2n=10 of this strain is clearly distinct from other species of the genus Paramacrobiotus, supporting the hypothesis that the strain represents a new species. Molecular analyses for the small and large ribosomal subunit (18S rDNA, 28S rDNA), the internal transcribed spacer 2 (ITS-2) and cytochrome C oxidase subunit I (COI) were also performed. The TYO strain is most similar in the analysed nuclear markers to Paramacrobiotus experimentalis Kaczmarek, Mioduchowska, Poprawa Roszkowska, 2020 and Paramacrobiotus sp. strain MG.002 (p-distances in 18S rDNA: 0.53%, 28S rDNA: 0.981.12%, and ITS-2: 9.9%), which corroborates with the overall morphological similarity between these taxa. Despite the close relationship between the TYO strain and P. experimentalis, the genetic species delimitation based on molecular analysis indicates that the TYO strain indeed is a distinct species. Therefore, this tardigrade is described here as Paramacrobiotus metropolitanus sp. nov.

Five Fabaceae Karyotype and Phylogenetic Relationship Analysis Based on Oligo-FISH for 5S rDNA and (AG3T3)3.

Most Fabaceae have nitrogen fixation abilities and are valuable forage and medicinal resources. However, cytogenetic data of many Fabaceae species are unclear. Karyotypes reveal cytological characteristics and are crucial to understanding the organization and evolution of chromosomes in species. Oligo-FISH can reveal genetic composition and karyotype variation patterns with rapid and efficient results. Karyotype analysis of five Fabaceae species by oligonucleotide probes showed that: Robinia pseudoacacia, karyotype formula 2n = 2x = 20m + 2sm, cytotype 2B, arm ratio 3.4821, eight chromosomes distributed 5S rDNA signal. The karyotype formula of Robinia pseudoacacia ‘idaho’ was 2n = 2x = 20m + 2sm, cytotype 1A, arm ratio 1.8997, and 5S rDNA signal was distributed on six chromosomes. Karyotype of Robinia pseudoacacia f. decaisneana 2n = 2x = 20m + 2sm, cytotype 1B, arm ratio 2.0787, the distribution of eight chromosomes with 5S rDNA signal. Karyotype formula of Styphnolobium japonicum 2n = 2x = 14m + 12sm + 2st, cytotype 2B, arm ratio 2.6847, two chromosomes have 5S rDNA signal. Amorpha fruticose karyotype 2n = 2x = 38m + 2sm, cytotype 1B, arm ratio 3.2058, four chromosomes possessed 5S rDNA signal. Both ends of all species’ chromosomes have (AG3T3)3 signals. The results of this study provide chromosome numbers and a physical map, contributing to the construction of the Oligo-FISH barcode and providing molecular cytogenetics data for Fabaceae.

Cytogenetic Analyses Detect Cryptic Diversity in Megaderma spasma from Malaysia

As many as 17 subspecies are currently recognized for the lesser false vampire, Megaderma spasma along its wide distributional range, from India to Southeast Asia. Cytogenetic studies have been so far restricted to specimens from only four geographical regions, namely Malaysia, Thailand, China and the Philippines. We examined the karyotypes of M. spasma medium from Peninsular Malaysia and M. s. trifolium from Malaysian Borneo with conventional and molecular cytogenetic techniques, using whole chromosome painting probes from the vespertilionid Myotis myotis and the hipposiderid Aselliscus stoliczkanus. Both Megaderma subspecies showed the same diploid chromosome number, 2n = 38, but differed concerning interstitial heterochromatic segments and number and shape of heterochromatic B chromosomes. Comparison with published data revealed that the Bornean specimens' karyotype best resembled the chromosomal set of the Chinese specimen. In contrast, a single male specimen from Malaysian Borneo showed a different karyotype (2n = 40) without B chromosomes and interstitial heterochromatin, indicating that it may represent a cryptic, yet unknown Megaderma species.