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.

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.

CHROMOSOMAL ANALYSIS OF EIGHT CULTIVARS IN THREE SPECIES OF CULTIVATED YAM (DIOSCOREA) SPECIES IN NIGERIA

The genus Dioscorea comprises of economically-important plant species known for their starch throughout the world; it is also a major source of food and income in Africa. The most important Dioscorea species cultivated and consumed in the West Africa belt include D. cayenensis, D. rotundata and D. alata. The plant materials used in this study were collected from Omu-Ekiti, Oye Local Government of Ekiti State, Nigeria using the On Farm Participatory Method (OFPM). Mitotic chromosome studies were carried out on three species viz, Dioscorea alata (‘ewura’), D. cayenensis (‘igangan’) and D. rotundata (‘areyingbakumo’, ‘gaungaun’, ‘ikumo’, ‘ogunmole’ and ‘sandpaper’). Mitotic chromosome studies were carried out on each of the cultivars using the root tip squash method made in modified Orcein stain (FLP-Orcein). Dormant tubers were cut to mini-setts and placed in carbonised rice husk for rooting. This study reports the basic chromosome number of x = 8, i.e. 2n = 4x =32 (D. alata), 2n = 4x =38 (D. rotundata) and 2n = 8x = 68 (D. cayenensis) for Dioscorea suggesting that both D. rotundata and D. cayenensis are aneuploids.

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 Variation in Eight Cultivars of Indian Dessert Banana (Musa acuminata L.) of Section Eumusa From Odisha, India

Banana (Musa spp.) cultivars especially dessert banana are important cash crop with high market demand all over the world as an integral part of the diet. The need for assessment of cytogenetic characters in Musa cultivars is inevitable as out of thousands of cultivars, cytogenetic characterization of most of them remains unresolved due to difficulties like small chromosome size, diversity in ploidy levels and high cultivar diversity which behave differently to standardized cytogenetic protocols. In this report, somatic chromosome number, detailed karyotype analysis including total chromosome length, volume, form percentage, Interphase Nuclear Volume (INV) were accessed on eight dessert type of Musa accessions from different places of Odisha. All the cultivars studied were found triploid (2n = 33) with a basic chromosome number of x=11. The karyotype formulae were assigned to each cultivar by grouping the chromosome according to their shared characteristics. The total chromosome length ranged from 54.95 µm in cv. Robusta to 81.5 µm in cv. Kathia with symmetric karyotype in all the studied cultivar. Karyotype formula revealed structural alteration of chromosome with Total Form percentage (TF%) variation from 35.65% in cv. Amritapani to 41.68% in cv. Patakpura that confirms more number of nearly median constricted chromosome as compared to sub-median chromosome. The total chromosome volume recorded from 10.78 µm3 in cv. Robusta to 15.99 µm3 in cv. Khatia and the INV varied from 1336.44 µm3 in cv. Dwarf Cavendish to 2048.37 µm3 in cv. Patakpura. The recorded structural variation might be due to differential genome specific condensation of chromosome. Chromosome length and volume found statistically significant among the cultivars.

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.

Somatic chromosome number and ploidy level in three Curcuma spp. from Bangladesh

Three Curcuma L. species were investigated cytogenetically which represent diversed staining pattern of heterochromatins in interphase nuclei and prophase chromosomes with orcein staining. Curcuma longa and C. caesia were found to possess 2n = 3x = 63 somatic chromosomes whereas 2n = 2x = 42 chromosome number in C. zedoaria is reported for the first time from Bangladesh. Total chromosome length recorded in C. longa, C. caesia and C. zedoaria were 145.08 ± 2.85 μm, 164.93 ± 4.29 μm and 97.78 ± 2.41 μm, respectively. This was the first attempt to measure the length of the chromosomes for these species. The experiment confirmed the basic chromosome number x = 21 with triploid (C. longa, C. caesia) and diploid (C. zedoaria) Curcuma plants. Polyploidy could be employed in the evolution and diversification of the genus Curcuma, which is an essential factor to characterize the species of this genus. Dhaka Univ. J. Biol. Sci. 30(2): 133-140, 2021 (July)

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.