Karyotype and genome size variation in white-flowered Eranthis sect. Shibateranthis (Ranunculaceae)

Comparative karyomorphological analyses of six out of the eight white-flowered species of Eranthis sect. Shibateranthis have been carried out. All studied specimens of E. byunsanensis, E. lobulata, E. pinnatifida, and E. stellata had a somatic chromosome number 2n = 16 with basic chromosome number x = 8. On the contrary, E. tanhoensis and E. sibirica had a basic chromosome number x = 7. The specimens of E. tanhoensis were diploid with 2n = 14, while the specimens of E. sibirica were polyploid with 2n = 42. Monoploid chromosome sets of the investigated diploid species had 4–5 metacentric chromosomes and 2–4 submetacentric/subtelocentric/acrocentric chromosomes. The highest level of interchromosomal asymmetry, estimated via CVCL, was found in E. byunsanensis and E. pinnatifida. The highest levels of intrachromosomal asymmetry (MCA) and heterogeneity in centromere position (CVCI) were found in E. lobulata and E. byunsanensis, while E. sibirica had the most symmetric karyotype. A multivariate PCoA analysis of basic karyotype parameters (2n, x, THL, CVCL, MCA, and CVCI) highlighted no overlap among species accessions, which was also confirmed by LDA. The average absolute monoploid DNA content (1Cx) of the 23 investigated samples of six Eranthis species varied from 9.26 ± 0.25 pg in E. sibirica to 15.93 ± 0.32 pg in E. stellata. Overall karyological affinity was highlighted between E. lobulata and E. stellata, on one side, and between E. byunsanensis and E. pinnatifida, on the other side. Interestingly, there was no significant correlation between total haploid (monoploid) chromosome length (THL) and 1Cx values in these species.

Chromosome and Genome Diversity in the Genus Trifolium (Fabaceae)

Trifolium L. is an economically important genus that is characterized by variable karyotypes relating to its ploidy level and basic chromosome numbers. The advent of genomic resources combined with molecular cytogenetics provides an opportunity to develop our understanding of plant genomes in general. Here, we summarize the current state of knowledge on Trifolium genomes and chromosomes and review methodologies using molecular markers that have contributed to Trifolium research. We discuss possible future applications of cytogenetic methods in research on the Trifolium genome and chromosomes.

Molecular Phylogeny of Trifolium L. Section Trifolium with Reference to Chromosome Number and Subsections Delimitation

The genus Trifolium is one of the largest genera of the legume family Fabaceae with ca. 255 species. The genus is divided into eight sections; the section Trifolium is a major section of the genus, comprising 73 species mainly distributed in the Mediterranean region. We used nuclear ribosomal DNA internal transcribed spacer (ITS) and morphological variation to reconsider the delimitation and phylogenetic relationships of species in the section Trifolium with reference to chromosomal variations. Bayesian analysis of ITS data delimited the species as three clades based on the analysis of ITS sequence and informative indels in combination with morphological variation. The phylogeny of the species by different analyses methods does not support their current delimitation in 17 subsections. The basic chromosome number x = 8 is the number for the genus Trifolium, from which x = 7, 6 and 5 were derived through successive aneuploidy events. With reference to the distribution of these numbers in the species of the section Trifolium, species in clade III and clade II are more evolved than species in clade I.

A karyotype study in two fish species belonging to Genus Neolissochilus found in Meghalaya, India

The karyomorphological study of two species of Mahseer belonging to the genus Neolissochilus, namely Neolissochilus hexagonolepis and N. hexastichus were carried out. The study revealed the basic chromosome number in both the Masheer species was observed to be 100. However, the karyotype formula number varied among the species. N. hexagonolepis had a diploid chromosome number of 42 metacentric (m), 20 submetacentric (sm), 8 subtelocentric (st) and 30 telocentric (t) and N. hexastichus had a karyotypic formula of 32 metacentric (m), 22 submetacentric (sm), 4 subtelocentric (st) and 42 telocentric (t). This finding removed taxonomic confusion due to the differences in the chromosome number, the morphology of the chromosomes and chromosome formula between the two fish species of the genus and helped in distinctive and unblemished identification of the two species belonging to the genus Neolissochilus from Meghalaya, though they have a morphological similarity.

Variation in Ribosomal DNA in the Genus Trifolium (Fabaceae)

The genus Trifolium L. is characterized by basic chromosome numbers 8, 7, 6, and 5. We conducted a genus-wide study of ribosomal DNA (rDNA) structure variability in diploids and polyploids to gain insight into evolutionary history. We used fluorescent in situ hybridization to newly investigate rDNA variation by number and position in 30 Trifolium species. Evolutionary history among species was examined using 85 available sequences of internal transcribed spacer 1 (ITS1) of 35S rDNA. In diploid species with ancestral basic chromosome number (x = 8), one pair of 5S and 26S rDNA in separate or adjacent positions on a pair of chromosomes was prevalent. Genomes of species with reduced basic chromosome numbers were characterized by increased number of signals determined on one pair of chromosomes or all chromosomes. Increased number of signals was observed also in diploids Trifolium alpestre and Trifolium microcephalum and in polyploids. Sequence alignment revealed ITS1 sequences with mostly single nucleotide polymorphisms, and ITS1 diversity was greater in diploids with reduced basic chromosome numbers compared to diploids with ancestral basic chromosome number (x = 8) and polyploids. Our results suggest the presence of one 5S rDNA site and one 26S rDNA site as an ancestral state.

Studies on Basic Chromosome Number, Ploidy Level, Chromosomal Association and Configuration and Meiotic Behavior in Mulberry (Morus Spp.)

Mulberry leaves are primary food for silkworm, Bombyx mori L. to feed silkworms and harvest quality silk cocoons. Mulberry belongs to family Moraceae and includes 60 species found distributed in both Hemisphere. In mulberry, chromosome numbers are varies from 2n = 28 to 22n = 308 (Diploid to Decosoploid) with ploidy level x to 22x. Based on chromosome numbers and meiotic behaviors x = 14 has been considered as basic chromosome numbers of the genus. In the present study, two diploids, two uneuploids, two triploids and two teteraploids mulberry varieties were selected for detailed chromosomal numbers and meiotic behaviors belongs to three species, namely Morus indica, Morus alba and Morus latifolia. Varieties, Vishaala and Kosen were diploids with 2n = 2x = 28 chromosomes and varieties Ber-S1 and S13 were uneuploids with 2n = 30 chromosomes belongs Morus indica. Varieties NAO Khurkul and KPG-1 were triploids with 2n = 3x = 42 chromosomes belongs to Moru alba and varieties Kokuso and Icheihei were tetraploids with 2n = 4x = 56 chromosomes. Diploids and uneuploids were showed normal meiosis with high pollen fertility and triploids and teteraploids were showed abnormal meiosis with low pollen fertility, due to virtue of higher ploidy level have been discussed in this chapter.

Genomic insights into the recent chromosome reduction and polyploidization of complex autopolyploid sugarcane S. spontaneum

S. spontaneum is a founding Saccharum species that contributes stress resistance to the genetic background of modern sugarcane cultivars. Here, we have assembled the autopolyploid S. spontaneum Np-X genome with ancestral form into 40 pseudo-chromosomes in 10 homologous groups, revealing the recent chromosome reduction and polyploidization that occurred in Saccharum. The paleo-duplicated chromosomal pairs exhibit functional redundancy in Saccharum and underwent fission followed by fusion accompanied by centromeric spreading around 0.80 million years ago (Mya) before evolving into their current forms with basic chromosome numbers x = 9 and x = 8 in S. spontaneum, likely in a stepwise manner. WGDs occurred independently in Saccharum species around 1.5 Mya. Highly diverse chromatin structures exist among homologous chromosomes despite their high collinearity, and the re-structuring of NpChr5 and NpChr8 might have suppressed switching of chromatin structure from inactive to active. Resequencing of 116 sugarcane accessions elucidated that the S. spontaneum originated from North India and that the basic chromosome numbers x = 8, x = 9, and x = 10 originated independently, indicating that recent chromosome reduction rather than polyploidization has driven the adaptive evolution of Saccharum. Our study provides genomic resources and suggests new directions for accelerating sugarcane improvement and advances our knowledge of the evolution of auto-polyploids.

In vitro propagation and cytological analysis of Sophora mollis Royle: an endangered medicinal shrub

Background Sophora mollis Royle (family Fabaceae, subfamily-Papilionaceae) is a multipurpose legume distributed in plains and foothills of the North-West Himalaya to Nepal and is facing high risk of extinction due to habitat loss and exploitation by the local people for its fuel and fodder values. Therefore, the present study was conducted to standardize a micropropagation protocol for Sophora mollis by using shoot tip explants and to study the meiotic chromosome count in the species. Results Multiple shoots were induced in shoot tip explants of Sophora mollis in Murashige and Skoog medium supplemented with different concentrations of cytokinins alone (BAP, TDZ, and Kinetin) and in combination with varying concentrations of NAA. MS medium supplemented with BAP (8.9 μM) was observed to be the optimal medium for multiple shoot induction and maximum 25.32 shoots per explant was obtained with average length of 4.5 ± 0.8 cm. In vitro developed shoots were transferred onto rooting media supplemented with different concentrations of auxin (IAA, IBA, and NAA). Maximum 86% rooting was observed in half-strength MS medium supplemented with 21.20 μM NAA with an average of 21.26 roots per culture. In vitro raised plantlets were adapted to greenhouse for better acclimatization and 60% plants were successfully transferred to the open environment. Based on the chromosome counts available from the literature and the current study, the species tend to show a basic chromosome number of x = 9. Conclusion The micropropagation protocol standardized can be helpful for the ex situ mass multiplication and germplasm conservation of the endangered species. Moreover, the ex situ conservation approach will be helpful in actively bridging the gap between ex situ and in situ approaches through the reintroduction of species in the wild. The cytological studies revealed the basic chromosome number x = 9 of the species.

Morphological and Cytological Characterization of Five Porterweed (Stachytarpheta) Selections

Porterweed (Stachytarpheta spp.), a member of the verbena family, is frequently used in pollinator gardens to attract butterflies. This study was conducted to assess the morphological features, pollen stainability and morphology, nuclear DNA content, and chromosome number of five porterweed selections. Coral porterweed (S. mutabilis), ‘Naples Lilac’ porterweed (S. cayennensis × S. mutabilis ‘Violacea’), and nettleleaf porterweed (S. cayennensis) had the largest plant heights. Flower number was significantly higher in nettleleaf porterweed, jamaican porterweed (S. jamaicensis), and U*J3-2 porterweed (S. cayennensis × S. jamaicensis), with an average of 65–72 flowers per inflorescence. Internode length and flower width of jamaican porterweed had much lower values than the other selections. Coral porterweed recorded the lowest pollen stainability with only 10.6% stainability, but it had the largest relative pollen production. ‘Naples Lilac’ porterweed had the highest DNA content with an average of 3.79 pg/2C, like jamaican porterweed with 3.73 pg/2C. Ploidy levels varied between selections, and the basic chromosome number was x = 28. Coral, jamaican, and ‘Naples Lilac’ porterweed had 2n = 6x = 168 chromosomes, first reported in this genus. These results provide a guide and a new tool to distinguish native and non-native porterweed and may aid future breeding toward the production of noninvasive cultivars.

Karyotype evolution of the genus Eranthis Salisb. (Ranunculaceae)

We have conducted comparative study of karyotypes for nine Eranthis Salisb. species: E. bulgarica (Stef.)Stef., E. hyemalis (L.) Salisb., E. longistipitata Regel (section Eranthis), E. byunsanensis B. Y. Sun, E. lobulata W. T.Wang, E. pinnatifida Maxim., E. sibirica DC., E. stellata Maxim., and E. tanhoensis Erst (section Shibateranthis). Thespecies-specifity of karyotypes was established for all species investigated. The chromosomes of each species weremedium or large in size (4–12 µm). Besides E. sibirica and E. tanhoensis, all the investigated specimens had diploidcytotypes with 2n = 16 and the basic chromosome number x = 8. Plants from five E. sibirica populations were tetraploidand hexaploid with x = 7, 2n = 28 and 2n = 42 respectively. Plants from seven E. tanhoensis populations were diploid withx = 7 and 2n = 14. Diploid karyotypes of Eranthis included 4–5 pairs of large equal-armed (metacentric) chromosomes,and 2–4 pairs of unequal-armed chromosomes belonging to different morphological types (submetacentric, subtelocentric,and acrocentric ones). We have revealed B chromosomes in root meristematic cells of E. lobulata and E. tanhoensis forthe first time. We suppose that the key developments in Eranthis karyotype`s evolution were pericentric inversions,polyploidy, and probably translocations.