Chromosomes in a hybrid zone of Israeli mole rars (Spalax, Rodentia)

Chromosomal novelties and the level of meiotic and mitotic abnormalities were studied in a hybrid zone between two chromosomally differentiated Spalax cytotypes of 2n = 58 and 2n = 52. These cytotypes differ by five Rb fusions, four centromeric shifts accompanied by heterochromatin deletion, one paracentric inversion, and the Y-chromosome reorganization. Among 149 specimens studied, 82 were hybrids with 64 different karyotypes ranging in diploid numbers from 2n = 50 to 2n = 60. Nine hybrid specimens were mosaics for the chromosome numbers due to occurrence of cell lines with different Robertsonian chromosome arrangements, and six specimens possessed variable number of B-chromosomes. Mosaicism of B-chromosomes was found also in meiotic cells however chromatid breaks and abnormal chromosome pairing during meiosis occurred very rarely. All these results imply some local genomic instability resulting in the spontaneous process of reversible Rb fusions.     

Cytological studies of Angiosperms (174 species) from District Kangra,Himachal Pradesh (India)

During the present study, 360 populations of 174 species belonging to 82 genera and 23 families of dicots have been cytologically worked out from different localities of District Kangra (Himachal Pradesh) between 550 and 3,800 m altitude. As many as 10 species including Berberis ceratophylla (2n = 28), Caltha alba (2n = 32), Corydalis meifolia (2n = 16), C. thyrsiflora (2n = 16), Impatiens reidii (2n = 14), Indigofera hamiltonii (2n = 16), Potentilla thomsonii (2n = 14), Sedum trifidum (2n = 36), Stellaria semivestita (2n = 26) and Viola canescens (2n = 12) have been cytologically worked out for the first time at world level. New intraspecific diploid cytotypes have been recorded in 11 species while new intraspecific tetraploid cytotypes are noticed in 15 species. B-chromosomes have also been reported in 3 species viz. Anemone obtusiloba, Clematis grata and Ranunculus diffusus for the first time at world level. The species of the area are active state of evolution, depicting heterogeneity in chromosome numbers involving polyploidy (48 species, 28.18 %) as well as irregular meiotic behavior. The meiotic abnormalities have been witnessed in 69 species (39.65 %) showing different types of irregularities such as cytomixis, interbivalant connections, unoriented bivalents, chromatin stickiness, chromosomal laggards and bridges as well as abnormal microsprogenesis. All these meiotic abnormalities lead to reduced pollen fertility and formation of variable sized pollen grains in most of these species.     

Cytomorphological diversity in some selected members of Poaceae from Parvati Valley in Kullu district of Himachal Pradesh,India

The present work includes detailed male meiotic studies on 46 species of grasses falling into 59 accessions from different localities of Parvati Valley in Kullu district of Himachal Pradesh in the altitudinal range of 1,100 to 2,750 m. All the species have been studied cytologically for the first time from the study area. The meiotic chromosome count of n = 14 for Calamagrostis emodensis is the first ever chromosome report. Three species, namely Agrostis alba (n = 21), Avena byzantina (n = 21) and Bromus inermis (n = 14) have been studied cytologically for the first time from India. New intraspecific diploid/polyploid cytotypes have been reported for Arthraxon serrulatus (2n = 4x = 32), Iseilema laxum (2n = 12x = 60), Digitaria albudens (2n = 8x = 72), Festuca kashmiriana (2n = 2x = 14) and Stipa orientalis (2n = 2x = 20). The existence of variable number of B-chromosomes (2n = 60 + 0-5B) has been reported for the first time in the 12x cytotype of Iseilema laxum. Secondary associations of chromosomes in the tetraploid cytotype of Cymbopogon martini (n = 20) indicated its secondary polyploid nature. As many as 18 species showed various meiotic anomalies such as the phenomenon of cytomixis involving inter PMC migration of chromatin material, chromatin stickiness, interbivalent connections, abnormal spindle activity, presence of bridges and laggards during anaphases and telophases and abnormal sporads. These meiotic abnormalities consequently yielded sterile and heterogeneous-sized fertile pollen grains. The polyploidy and aneuploidy have played an active role in the evolution of grasses.     

CHROMOSOMAL EVOLUTION IN RHOGEESSA (CHIROPTERA: VESPERTILIONIDAE): POSSIBLE SPECIATION BY CENTRIC FUSIONS

Within the Rhogeessa tumida-parvula complex, there are seven cytotypes involving diploid numbers of 30, 32, 34, 42, 44, and 52. Based on G-band analysis, the 30, 32, 34, and 44 forms differ from each other by centric fusions (13 different fusions were identified). Two 2n = 32 samples (one from Belize and one from Nicaragua) having essentially identical standard karyotypes differed from each other by eight fusion events. Cytotypes are allozymically distinct; however, other studies indicate that the cytotypes are morphologically very similar if not indistinguishable (LaVal, 1973; Baker, 1984). G-band chromosomal data are best interpreted as indicating that, within this complex, several species exist which should be maintained by a post-mating isolating mechanism resulting from meiotic problems in F₁ individuals. Mus musculus (Capanna et al., 1977; Capanna, 1982; White, 1978) and the Rhogeessa tumida-parvula complex are unusual among mammalian species thus far studied in their pattern of chromosomal variation and presumptive mode of speciation. Strong evidence exists suggesting that in both groups populations differing by multiple centric fusions are biological species. It appears that the population and reproductive biology of Mus and Rhogeessa are quite different and any model that attempts to explain the mode of speciation in these two diverse taxa, must be compatible with their different biological characteristics.     

Karyological knowledge of the Italian vascular flora as inferred by the analysis of “Chrobase.it”

Abstract

Chromosome number knowledge of the Italian vascular flora is stored in the online database Chrobase.it, which includes 6723 records, referable to 3428 taxa, 2799 accepted species and subspecies (about 35% of the national flora), and 3410 different chromosome countings (cytotypes). Appropriate queries to Chrobase.it allowed us to calculate mean, modal and median chromosome numbers for the Italian vascular flora, for geographical subgroups (islands, south, centre, north) and for selected orders, families and genera. Chromosome number data were available for 41 out of 55 orders (74%) and 107 out of 428 families (67%), represented by 664 out of 1297 genera (51%). The most studied administrative regions are Sicily (844 taxa), Tuscany (592 taxa), and Sardinia (390 taxa), while the most studied families are Asteraceae (465 taxa), Fabaceae (266 taxa), Brassicaceae (158 taxa), and Poaceae (144 taxa). Chromosome numbers range from 2n = 6, occurring in several species of Hypochaeris (Asteraceae), to 2n = 240, occurring in Ophioglossum (Ophioglossaceae), Dryopteris (Dryopteridaceae) and Arenaria (Caryophyllaceae) (mode is 2n = 18, and median is 2n = 24). Chromosome number variability was analyzed by frequencies (linear plots) and ANOVA, resulting in significant differences among geographical groups (mean chromosome number increasing from islands-south to centre-north) and selected taxa. B-chromosomes occur in 5.3% of data (148 taxa) and their number is not significantly different among geographical areas, while they occur only in 14 orders, 17 families, and 56 genera. The number of B-chromosomes ranges from 1 to 13 (mode = 1, median = 2).     

Cytological evaluation of Apiaceae Lindl. from Western Himalayas

The present paper deals with cytological studies on 31 populations covering 17 species belonging to 10 genera of Apiaceae from Western Himalayas. The chromosome numbers in the two species as Chaerophyllum capnoides (n = 11) and Heracleum brunonis (n = 11), along with additional cytotypes for Pimpinella acuminata (n = 9) and Sium latijugum (n = 12) have been reported for the first time on world-wide basis. The genus Pleurospermum, although cytologically worked out earlier from outside India, its species densiflorum (n = 11) makes first representation of the genus from India. Besides, the chromosome number in Chaerophyllum aromaticum (n = 11) have been worked out for the first time from India. The course of meiosis varies from normal to abnormal in different populations of Chaerophyllum villosum, Pimpinella achilleifolia and Sium latijugum while abnormal meiotic course has been observed in all the studied populations of Chaerophyllum acuminatum, C. aromaticum, C. capnoides, Pimpinella acuminata, P. diversifolia, Pleurospermum densiflorum and Vicatia coniifolia. Such taxa are marked with meiotic abnormalities in the form of cytomixis, chromatin stickiness, formation of laggards and bridges resulting into abnormal microsporogenesis. The occurrence of structural heterozygosity has been recorded in the Chaerophyllum acuminatum and C. aromaticum. The effect of these abnormalities is clearly seen on the pollen size and fertility.     

Ecology of two cytotypes ofButomus umbellatus I. Karyology and breeding behaviour

Chromosome numbers were counted inButomus umbellatus from 99 localities in both Czech and Slovak Republics and the karyotype morphology was studied. Basic chromosome sets are asymmetrical and uniform among diploids (2n=26) and triploids (2n=39). Diploids occur less frequently than triploids in this region. Their clonal populations are usually fertile owing to self-compatibility. The clonal populations of triploids are selfincompatible and usually sterile. Thus, the different self-compatibility is the main biological character distinguishing diploids from triploids. Pollen of triploids is viable in spite of meiotic irregularities in pollen mother cells (PMCs). Hybridization both between cytotypes and between the different triploid genets may take place, if they occur sympatrically. Offspring having diverse chromosome numbers between diploid and triploid level can originate as the consequence of such hybridization.     

Cytogenetic variations in eight species of Saussurea DC. (Asteraceae,Cardueae) from Northwest Himalaya

The genus Saussurea comprises many species with highly medicinal, religious or other economical values. The chromosome number in the genus ranges from 2n = 24 to 108, based on x = 12, 13, 16, or 17, but the primary base number is still not clear. The present meiotic study cover 8 species (14 populations) of Saussurea from the Northwest Himalayas, and add new or varied cytotypes, as an attempt to solve the enigmatic cytogenetic variations in the genus. The chromosome numbers in Saussurea auriculata (n = 16) is new to the world, while S. costus (n = 17), S. jacea (n = 17) and S. roylei (n = 17) reveal the varied cytotypes at world level. Besides, S. taraxicifolia (n = 16) is a first ever report for Indian populations. The meiotic studies on different populations reveal a substantial amount of meiotic abnormalities in the form of chromosome stickiness, un-oriented bivalents, cytomixis, laggards, etc. leading to the meiocytes with less (aneuploidy) or more (polyploidy) chromosome numbers. These abnormalities may produce unreduced pollen grains and adversely affect pollen viability. The high number of these meiotic abnormalities may be responsible for the chromosome number variation in the genus.     

Cytology of the genus Artemisia (Anthemidae,Asteraceae) in the Western Himalayas

The present study revealed the varied frequency of natural chromosomal abnormalities in 13 populations pertaining to 9 species of the genus Artemisia L. from different localities of Himachal Pradesh (Western Himalaya). Intraspecific chromosome variability has been reported for the first time on worldwide basis in Artemisia vestita (2n = 2x = 36) and from India in A. macrocephala (2n = 2x = 18) and A. scoparia (2n = 2x = 36). Besides, B-chromosomes have been reported here for the first time in A. nilagirica and A. roxburghiana. Most of the populations show anomalous meiotic behaviour resulting in cytomixis, chromosomal stickiness, unoriented bivalents, formation of laggards and bridges which leads to abnormal microsporogenesis, and production of heterogeneous-sized fertile pollen grains along with reduced pollen fertility.     

REDUCED GENE FLOW AT PERICENTROMERIC LOCI IN A HYBRID ZONE INVOLVING CHROMOSOMAL RACES OF THE HOUSE MOUSE MUS MUSCULUS DOMESTICUS

The West European house mouse, Mus musculus domesticus, is a particularly suitable model to investigate the role of chromosomal rearrangements in reproductive isolation. In fact, it exhibits a broad range of chromosomal polymorphism due to Robertsonian (Rb) fusions leading to various types of contact zones between different chromosomal races. In the present study, we analyzed a parapatric contact in central Italy between the Cittaducale chromosomal race (CD: 2n= 22) and the surrounding populations with standard karyotype (2n= 40) to understand if Rb fusions play a causative role in speciation. One hundred forty‐seven mice from 17 localities were genotyped by means of 12 microsatellite loci. A telomeric and a pericentromeric locus situated on six chromosome arms (four Rbs and one telocentric) were selected to detect differences in the amount of gene flow for each locus in different chromosomal positions. The analyses performed on the two subsets of loci show differences in the level of gene flow, which is more restricted near the centromeres of Rb chromosomes. This effect is less pronounced in the homozygotes populations settled at the border of the hybrid zone. We discuss the possible cause of the differential porosity of gene flow in Rbs considering “hybrid dysfunctions” and “suppressed recombination” models.     

Detailed cytomorphological investigations through male meiosis of polypetalous plants from the Kashmir Himalaya

Cytomorphological studies were made on 322 populations of 150 species/153 taxa belonging to 79 genera and 23 families of flowering plants. Out of these, 14 species were cytologically worked out for the first time on a worldwide basis along with B-chromosomes (0–1B) recorded for the first time in Agrimonia eupatoria. Similarly, 29 species were reported with varied chromosomal reports on a worldwide basis. The chromosome numbers ranged from 2n = 10 to 2n = 96 and posed high amount of intraspecific genetic diversity when seen in relation to other reports for the worked out species in light of previous chromosomal databases on India and world basis. The different levels of ploidy in these species/taxa vary from 2x to 12x with the overall polyploidy being 27.45 % and predominant in members belonging to the genera Ranunculus and Epilobium as well as families Ranunculaceae and Rosaceae. In the taxa showing anomalous meiotic behavior, cytomixis is found to be quite common and associated with various meiotic abnormalities in the meiocytes, which ultimately leads to pollen sterility and formation of variable sized pollen grains.     

Extensive chromosome number variation in Aster ageratoides var. pendulus (Asteraceae)

Aster ageratoides var. pendulus, a recently described taxon, is endemic to Mt Hupingshan of north‐western Hunan, China. Field observations and collections were made from the only known population. Root‐tip squashes were used to determine the chromosome numbers of 96 plants and 61 seedlings from the achenes of eight sample plants. The results show that var. pedulus is a swarm of 30 cytotypes with nearly continuous chromosome numbers from 2n = 60 to 2n = 92. Chromosome numbers of 61 seedlings vary from 2n = 61 to 2n = 91, belonging to 18 cytotypes. The chromosome number variation of var. pendulus is highly unusual not only in the A. ageratoides polyploid complex but also in angiosperms. Such an enormous continuous variation of chromosome numbers could have arisen by the combined effect of hybridization, recent origin and high levels of polyploidy. © 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 165 , 378–387.     

Genetic control of synapsis and recombination in Lolium amphidiploids

Homologous bivalent formation in amphidiploids of Lolium is promoted during meiosis by diploidising genes carried by A-chromosomes and by supernumerary B-chromosomes. The site and mode of action of these diploidising factors were investigated by comparing the relative frequencies of pairing configurations at meiotic prophase and metaphase I in several different hybrid genotypes. The results indicate that diploidising genes act predominatly by increasing the stringency of synapsis at early stages of meiotic prophase. By contrast, B-chromosomes appear to promote bivalent formation by ensuring that homoeologously paired chromosome segments within multivalents do not crossover. The results show that the additive effects of diploidising genes and B-chromosomes are to a certain extent separable in terms of their mode of action and timing during meiosis.     

Chromosome numbers in some cacti of western North America

Documented chromosome numbers and meiotic behavior were recorded for 23 taxa of 18 species of Cactaceae of south-western United States and adjacent Mexico. All taxa are diploid (n = 11) or polyploid (n = 22, 33, 44), and with regular meiotic behavior.     

The role of triploids in the origin and evolution of polyploids of <Emphasis Type="Italic">Turnera sidoides</Emphasis> complex (Passifloraceae,Turneroideae)

Triploids can play an important role in polyploid evolution. However, their frequent sterility is an obstacle for the origin and establishment of neotetraploids. Here we analyzed the microsporogenesis of triploids (x?=?7) and the crossability among cytotypes of Turnera sidoides, aiming to test the impact of triploids on the origin and demographic establishment of tetraploids in natural populations. Triploids of T. sidoides exhibit irregular meiotic behavior. The high frequency of monovalents and of trivalents with non-convergent orientations results in unbalanced and/or non-viable male gametes. In spite of abnormalities in chromosome pairing and unbalanced chromosome segregation, triploids are not completely sterile and yielded up to 67% of viable pollen. Triploids that originated by the fusion of 2n?×?n gametes of the same taxon showed more regular meiotic behavior and higher fertility than triploids from the contact zone of diploids and tetraploids or triploids of hybrid origin. The reproductive isolation of T. sidoides cytotypes of different ploidy level is not strict and the ‘triploid block’ may be overcome occasionally. Triploids of T. sidoides produce diploid and triploid progeny suggesting that new generations of polyploids could originate from crosses between triploids or from backcrosses with diploids. The capability of T. sidoides to multiply asexually by rhizomes, would enhance the likelihood that a low frequency of neopolyploids can be originated and maintained in natural populations of T. sidoides.     

CYTOGENETIC ANALYSIS OF CHROMOSOMAL INTERMEDIATES FROM A HYBRID ZONE BETWEEN TWO CHROMOSOME RACES OF THE SCELOPORUS GRAMMICUS COMPLEX (SAURIA,PHRYNOSOMATIDAE)

Underdominance for chromosomal rearrangements is the central assumption of several models of chromosomally based speciation including the cascade model, proposed for the Sceloporus grammicus complex. Several cytotypes of the S. grammicus complex hybridize at localities in central México. A hybrid zone between two of the most chromosomally divergent races (= cytotypes) of S. grammicus (F5, 2n = 34 and FM2, 2n = 44–46) was examined to assess the meiotic effects of heterozygosity at multiple chromosomes. Meiosis was examined in males heterozygous for “simple” Robertsonian fissions at chromosomes 1, 3, 4, and 6 and/or a pericentric inversion at chromosome 4. Analysis of synaptonemal complexes and chromosomal configurations at diakinesis showed trivalent formation in fission heterozygotes and heterosynapsis (lack of reverse-loop formation) in an inversion heterozygote. Analysis of metaphase II configurations revealed primarily balanced segregation and low levels of nondisjunction regardless of chromosomal background. The lack of underdominance associated with “simple” fission heterozygosity in this narrow hybrid zone contradicts the key premise of most chromosomally based models of speciation.     

Evolution and taxonomic split of the model grass Brachypodium distachyon

Background and Aims

Brachypodium distachyon is being widely investigated across the world as a model plant for temperate cereals. This annual plant has three cytotypes (2n =  10, 20, 30) that are still regarded as part of a single species. Here, a multidisciplinary study has been conducted on a representative sampling of the three cytotypes to investigate their evolutionary relationships and origins, and to elucidate if they represent separate species.

Methods

Statistical analyses of 15 selected phenotypic traits were conducted in individuals from 36 lines or populations. Cytogenetic analyses were performed through flow cytometry, fluorescence in situ hybridization (FISH) with genomic (GISH) and multiple DNA sequences as probes, and comparative chromosome painting (CCP). Phylogenetic analyses were based on two plastid (ndhF, trnLF) and five nuclear (ITS, ETS, CAL, DGAT, GI) genes from different Brachypodium lineages, whose divergence times and evolutionary rates were estimated.

Key Results

The phenotypic analyses detected significant differences between the three cytotypes and demonstrated stability of characters in natural populations. Genome size estimations, GISH, FISH and CCP confirmed that the 2n = 10 and 2n = 20 cytotypes represent two different diploid taxa, whereas the 2n = 30 cytotype represents the allotetraploid derived from them. Phylogenetic analysis demonstrated that the 2n = 20 and 2n = 10 cytotypes emerged from two independent lineages that were, respectively, the maternal and paternal genome donors of the 2n = 30 cytotype. The 2n = 20 lineage was older and mutated significantly faster than the 2n = 10 lineage and all the core perennial Brachypodium species.

Conclusions

The substantial phenotypic, cytogenetic and molecular differences detected among the three B. distachyon sensu lato cytotypes are indicative of major speciation processes within this complex that allow their taxonomic separation into three distinct species. We have kept the name B. distachyon for the 2n = 10 cytotype and have described two novel species as B. stacei and B. hybridum for, respectively, the 2n = 20 and 2n = 30 cytotypes.     

Chromosome numbers,characterization of chromosomal pairing during meiosis,origin and natural propagation in polyploid cytotypes (4x, 5x and 6x) of Agrimonia eupatoria L. (Rosaceae) in northwest Himalayas (India)

Despite the presence of intraspecific polyploidy (2x, 4x, 5x and 6x) in Agrimonia eupatoria, origin of these cytotypes has never been addressed adequately. The aim of the present study was to record the original chromosome counts and characterize chromosomal pairing during meiosis and microsporogenesis in the 5x cytotype, and discussing the hypothesis regarding the possible origin of polyploid cytotypes (4x, 5x and 6x) in the species. The geographical distribution pattern of cytotypes in the Indian Himalayas and elsewhere has also been analyzed. The present meiotic analysis revealed three chromosomes counts, the tetraploid (2n?=?4x?=?56), the pentaploid (2n?=?5x?=?70) and the hexaploid (2n?=?6x?=?84) cytotypes based on x?=?14. Meiotic course was perfectly normal in the 4x and 6x cytotypes resulting into high pollen fertility (94–100 %). Meiotic course in the imbalanced 5x cytotype has been found to be irregular characterized by the presence of high frequency of univalents at diakinesis and metaphase-I. Abnormal meiotic course contributed towards high pollen sterility (74–88 %). Even the apparently fertile/stained pollen grains were of irregular shape and of heterogeneous sizes. Meiotic behaviour of the 5x cytotype is like typical of allopolyploid. Individuals of 5x cytotype did not produce seeds and propagate vegetatively (root suckers) while 4x and 6x cytotypes exploited sexual (seeds) as well as vegetative means for propagation. Chromosomal pairing in pentaploid cytotype is like typical of an allopolyploid and we assume that it might have originated owing to natural inter-cytotype hybridization between 4x and 6x cytotypes in a mixed population. Analysis of geographical distribution pattern of cytotypes shows that Indian Himalayas represent the most cytotype-diverse region for A. eupatoria with the existence of all the four cytotypes (2x, 4x, 5x, 6x). This shows the dynamic nature of the species at chromosomal level in this part of the world.     

Reproductive isolation in a nascent species pair is associated with aneuploidy in hybrid offspring

Speciation may occur when the genomes of two populations accumulate genetic incompatibilities and/or chromosomal rearrangements that prevent inter-breeding in nature. Chromosome stability is critical for survival and faithful transmission of the genome, and hybridization can compromise this. However, the role of chromosomal stability on hybrid incompatibilities has rarely been tested in recently diverged populations. Here, we test for chromosomal instability in hybrids between nascent species, the ‘dwarf’ and ‘normal’ lake whitefish (Coregonus clupeaformis). We examined chromosomes in pure embryos, and healthy and malformed backcross embryos. While pure individuals displayed chromosome numbers corresponding to the expected diploid number (2n = 80), healthy backcrosses showed evidence of mitotic instability through an increased variance of chromosome numbers within an individual. In malformed backcrosses, extensive aneuploidy corresponding to multiples of the haploid number (1n = 40, 2n = 80, 3n = 120) was found, suggesting meiotic breakdown in their F₁ parent. However, no detectable chromosome rearrangements between parental forms were identified. Genomic instability through aneuploidy thus appears to contribute to reproductive isolation between dwarf and normal lake whitefish, despite their very recent divergence (approx. 15–20 000 generations). Our data suggest that genetic incompatibilities may accumulate early during speciation and limit hybridization between nascent species.     

Chromosome counts in Clibadium (Compositae,Heliantheae) from Latin America

New meiotic chromosome counts are reported from 86 populations from Mexico to Ecuador of 13 species ofClibadium (Compositae, Heliantheae). These, plus previous reports, yield a total of 120 counts from 20 of the approximately 39 species recognized in the genus (Arriagada, in prep.). All counts aren=16 with a few fragments or B-chromosomes observed sporadically in some populations. One population, containing what appear morphologically to be interspecific hybrids betweenC. mexiae andC. microcephalum, showed one tetravalent and fragments. Species in both historical sections (Clibadium andTrixidium, created by Candolle and maintained by Schulz) and from the five sections to be proposed in a new classification of the genus (Arriagada, in prep.) have now been counted. The uniformity of chromosome numbers withinClibadium is correlated with observed allopatric distribution of closely related taxa.