Linked-Read Sequencing of Eight Falcons Reveals a Unique Genomic Architecture in Flux

Falcons are diverse birds of cultural and economic importance. They have undergone major lineage specific chromosomal rearrangements, resulting in greatly-reduced chromosome counts relative to other birds. Here, we use 10X Genomics linked reads to provide new high-contiguity genomes for two gyrfalcons, a saker falcon, a lanner falcon, three subspecies of peregrine falcons, and the common kestrel. Assisted by a transcriptome sequenced from 22 gyrfalcon tissues, we annotate these genomes for a variety of genomic features, estimate historical demography, and then investigate genomic equilibrium in the context of falcon-specific chromosomal rearrangements. We find that falcon genomes are not in AT-GC equilibrium with a bias in mutations towards higher AT content; this bias is predominantly driven by, but not dependent on, hypermutability of CpG sites. Small indels and large structural variants were also biased towards insertions rather than deletions. Patterns of disequilibrium were linked to chromosomal rearrangements: falcons have lost GC content in regions that have fused to larger chromosomes from microchromosomes and gained GC content in regions of macrochromosomes that have translocated to microchromosomes. Inserted bases have accumulated on regions ancestrally belonging to microchromosomes, consistent with insertion-biased gene conversion. We also find an excess of interspersed repeats on regions of microchromosomes that have fused to macrochromosomes. Our results reveal that falcon genomes are in a state of flux. They further suggest that many of the key differences between microchromosomes and macrochromosomes are driven by differences in chromosome size, and indicate a clear role for recombination and biased gene conversion in determining genomic equilibrium.

Chloroplast phylogenomics in Camelina (Brassicaceae) reveals multiple origins of polyploid species and the maternal lineage of C. sativa

The genus Camelina (Brassicaceae) comprises 7–8 diploid, tetraploid, and hexaploid species. Of particular agricultural interest is the biofuel crop, C. sativa (gold-of-pleasure or false flax), an allohexaploid domesticated from the widespread weed, C. microcarpa. Recent cytogenetics and genomics work has uncovered the identity of the parental diploid species involved in ancient polyploidization events in Camelina. However, little is known about the maternal subgenome ancestry of contemporary polyploid species. To determine the diploid maternal contributors of polyploid Camelina lineages, we sequenced and assembled 84 Camelina chloroplast genomes for phylogenetic analysis. Divergence time estimation was used to infer the timing of polyploidization events. Chromosome counts were also determined for 82 individuals to assess ploidy and cytotypic variation. Chloroplast genomes showed minimal divergence across the genus, with no observed gene-loss or structural variation. Phylogenetic analyses revealed C. hispida as a maternal diploid parent to the allotetraploid Camelina rumelica, and C. neglecta as the closest extant diploid contributor to the allohexaploids C. microcarpa and C. sativa. The tetraploid C. rumelica appears to have evolved through multiple independent hybridization events. Divergence times for polyploid lineages closely related to C. sativa were all inferred to be very recent, at only ~65 thousand years ago. Chromosome counts confirm that there are two distinct cytotypes within C. microcarpa (2n = 38 and 2n = 40). Based on these findings and other recent research, we propose a model of Camelina subgenome relationships representing our current understanding of the hybridization and polyploidization history of this recently-diverged genus.

Chromosome numbers of eight Carex taxa in Korea (Cyperaceae)

In the flora of Korea, Carex L. is one of the most species-rich genera. Among nearly 157 Carex taxa, less than 30 have had their chromosome numbers reported. We report the meiotic chromosome numbers of eight Carex taxa from Korean populations, which include the first count for C. accrescens Ohwi (n = 37II) and the first chromosome investigations of Korean populations for three taxa: C. bostrychostigma Maxim. (n = 22II), C. lanceolata Boott (n = 36II), and C. paxii Kük. (n = 38II). In most species, chromosome counts observed in the study are included within the variation ranges of previous chromosome numbers. However, C. bostrychostigma Maxim. (n = 22II) and C. planiculmis Kom. (n = 29II) are assigned new chromosome numbers. Carex is known to have holocentric chromosomes, lacking visible primary constrictions and exhibiting great variance in its chromosome number. Further investigations of the diversity of Carex chromosomes will provide basic information with which to understand the high species diversity of the genus.

Cytogenetic, chromosome count optimization and automation of Neolamarckia cadamba (Rubiaceae) root tips derived from in vitro mutagenesis

Chromosome count is the only direct way to determine the number of chromosomes of a species. This study is often considered trivial that seldom described and discussed in detail. Therefore, it is inevitable that the chromosome count protocol should be revised and revisited before it becomes obliterated. In the present study, we encountered challenges in obtaining a clear micrograph for the chromosome count of active mitotic cells of Neolamarckia cadamba (Roxb.) Bosser (Rubiaceae) root tips. Several obstacles were determined through micrograph observation, such as existing unwanted particles in cells, poor chromosome staining and chromosome clumping. To overcome these, root tip types, staining methodologies, squashing methods were among the factors assessed to obtain clear micrographs. The chromosome counts of N. cadamba under optimized procedure showed 2n = 44 chromosomes. We also apply digital technology in chromosome counts, such as online databases and graphic software that are open source and freely accessible to the public. Only basic laboratory equipment and chemicals were used throughout the study, thus making this study economical and applicable in a basic laboratory. The availability of online digital software and databases provide open-source platforms that will ease the efforts in chromosome count.

Chromosome counts of eight Iranian endemic species of Nepeta L. (Lamiaceae)

In this survey, the chromosome counts of eight Nepeta L. species were investigated and the karyotypic diversity among these species was studied. The examined species belong to N. cephalotes Boiss. species group, namely N. eremokosmos Rech.f., N. gloeocephala Rech. f., cephalotes Boiss., N. pungens (Bunge) Benth., N. ispahanica Boiss., N. mahanensis Jamzad & Simonds, N. hormozganica Jamzad and N. denudata Benth. collected from different habitats in Iran. The ploidy levels, karyotype formula, chromosome length range, total karyotype length, several karyotype asymmetries values and Stebbins classification were determined in this study. Results showed the same chromosome number, 2n = 2x= 18 for all studied species. The basic chromosome number for the above mentioned species are x = 9. Also, the smallest chromosome length is 1.02 μm in N. mahanensis. The largest chromosome length is 2.3 μm in N. ispahanica. The chromosomes of species were metacentric or submetacentric. According to the Stebbins classification, these species were located into three classes 1A, 2A and 3A. The chromosome numbers for six of studied species are reported here for the first time.

Karyotype analysis of Aeluropus species (Poaceae)

Aeluropus, a member of Poaceae subfam. Chloridoideae, includes six species, three of which occur in Iran. They are perennial halophytes of deserts and coastal marshlands of Iran. The genus is considered as a rich genetic source for gene manipulation and using it for crop improvement. Previous studies showed that members of Chloridoideae have small chromosomes and the base chromosome number n = 10. There are few chromosome records for Aeluropus species. Somatic metaphases of seven populations of three Aeluropus species were studied. The first chromosome counts (2n = 20) based on Iranian material for three species, A. macrostachyus, A. littoralis and A. lagopoides, are concordant with previous records outside Iran; mitotic number for A. macrostachyus is recorded here for the first time.

Gynogenetic diploids, tetraploids, or octoploids, and a path to polyploidy in Anuran Amphibians

Unreduced gametes have been implicated in the evolution of polyploid species of plants and animals and are normally produced by female anuran amphibians. Such eggs may initiate the evolution of polyploid species that have independently arisen in several anuran families. Polyploid females could also produce unreduced eggs that might lead to species with higher ploidy levels or their eggs may develop gynogenetically to reduce the ploidy level. Diploid Hyla chrysoscelis (2n=24) and tetraploid H. versicolor (4n=48) are sibling cryptic species of North American Grey Treefrogs. Artificial crosses using H. versicolor females and genetically distant diploid males were performed to produce haploid H. versicolor and to assess the production of unreduced eggs in this tetraploid species. Gynogenetic diploid (haploid H. versicolor), allotriploid, gynogenetic tetraploid, allopentaploid, autohexaploid, and gynogenetic octoploid tadpoles were confirmed using chromosome counts from tadpole tail tip squashes. Transformation and survival of the different ploidies varied. Gynogenetic diploids transformed but expressed aspects of the haploid syndrome and died before or shortly after transformation.

NEW CHROMOSOME COUNTS OF ASIAN COSTACEAE AND INITIAL INSIGHTS INTO THE GENOME EVOLUTION OF THE FAMILY

Chromosome counts were obtained from six species of Costaceae from Asia. Our count of 2n = 18 for Cheilocostus speciosus confirms previous counts, and the other five counts have been made for the first time (Cheilocostus borneensis, Cheilocostus globosus, Cheilocostus sopuensis, Costus muluensis and Paracostus sp.). These chromosome counts reveal two somatic numbers, 2n = 18 and 2n = 36, of which the former is a new diploid number for the genus Paracostus. A comprehensive review of existing counts was conducted through literature and database searches. Mapping of these on a published comprehensive phylogenetic tree suggests that the diploid count of 2n = 18 is probably ancestral in the Costaceae, with repeated parallel evolution of tetraploidy and one case of octoploidy. The existence of triploid counts in several lineages harbouring polyploids suggests that diploids and tetraploids may exist in close proximity, and that crosses or meioticirregularities may lead to triploid genotypes occurring frequently.