Cytological studies on meiotic configurations of intraspecific aneuploids ofCarex blepharicarpa (Cyperaceae) in Japan

Chromosome configurations at meiotic metaphase I ofCarex blepharicarpa were determined for 245 individuals collected from 11 localities in the Chugoku District of Japan. Nine intraspecific aneuploids, 2n=26–33 and 41, were found. The most common diploid number was 29, and found in 73 individuals. No clear geographical pattern was suggested by a distribution of these aneuploids. A consecutive series of chromosome numbers from 2n=26 to 32 was found in two populations from Okayama Prefecture. At meiotic metaphase I, univalents and multivalents were found, and one to three trivalents were observed in each aneuploid. Heteromorphic chain trivalents comprising large, medium and small chromosomes were prevalent. Homomorphic trivalents, which are thought to be originated from chromosome duplications caused by unreduced gametes, were found in only one individual with 2n=41. The high frequency of heteromorphic trivalents in this species indicates that most aneuploidy probably results from fission and/or fusion of chromosomes.     

Cytogeographical study of four aneuploids ofCarex oxyandra Kudo in Japan

Chromosome numbers were determined for 342 clones ofCarex oxyandra collected from 35 localities in Hokkaido, Honshu, Shikoku and Kyushu, Japan. Four intraspecific aneuploids, 2n=18, 20, 24 and 26, were found. In meiotic division, only bivalent chromosomes were observed in all clones at metaphases I and II, suggesting that the aneuploids are established gamodemes. In the mitotic metaphase chromosomes, trimodal variation in chromosome length was observed. The 2n=26 clones found on Mt. Hiko had two particularly small chromosomes. The cytodemes with higher number of chromosomes are distributed in more southern areas of Japan.Carex oxyandra, therefore, accompanied with chromosome fragmentations, might spread the geographical distribution to the southern parts. The morphological characters of leaves, spikes, scales, perigynia and nuts were similar among the four cytodemes, except for the small leaves on plants from Yaku Island.     

Biosystematic studies on the genusIsoetes in Japan I. Variations of the somatic chromosome numbers

The somatic chromosome number of three Japanese species ofIsoetes, I. asiatica, I. japonica andI. sinensis, was determined in 199 individuals from 49 populations. The chromosome number ofI. asiatica was 2n=22, confirming previous reports. However,I. japonica andI. sinensis displayed a diversity in chromosome number. Six cytotypes, 2n=66, 67, 77, 87, 88 and 89, were found inI. japonica; 2n=67, 87, 88 and 89 are new counts in the genusIsoetes. The plants with 2n=66 were the most frequent (72% of total individuals examined) and were distributed throughout Honshu and Shikoku. The plants with 2n=88 occurred in western Honshu and a limited region in northeastern Honshu where the plants with 2n=77 were also found. In contrast, four cytotypes, 2n=44, 65, 66 and 68, were found inI. sinensis. The chromosome numbers ofI. sinensis were reported here for the first time. The plants with 2n=44 occurred only in Kyushu, while the plants with 2n=66 were found throughout a large area of western Japan.     

Karyotypic changes in potato plants regenerated from protoplasts

Over two hundred plants were regenerated from shoot-culture derived proto-plasts of potato (Solanum tuberosum L. cv. Majestic). Some had grossly aberrant phenotypes but the majority were similar to, or indistinguishable from normal control Majestic. Cytological examination showed that on average, 57% of the regenerants had the normal chromosome number (2n=4x=48). The remainder were aneuploids and fell into two classes in approximately equal numbers. The first class was limited at about the euploid level (ie, 2n=44–49). The second class contained plants with higher chromosome numbers ranging from 2n=73 to the octaploid level (2n=8x=96). The overall results represent an improvement over our earlier studies on chromosome variation in protoplast-derived potato plants. In addition, three cases of structural chromosome variation were observed.     

Variable karyotype with B-chromosomes in Bellevalia saviczii (Liliaceae)

Bellevalia saviczii (Liliaceae), a hexaploid (2n = 6x = 24), was collected from mountain slopes in the Shiraz valley, southern Iran. The basic karyotype consists of one long metacentric chromosome, a slightly shorter acrocentric, and two shorter submetacentrics. In 32% of the plants, one and sometimes two of the acrocentrics (IIa) appeared with a pericentric inversion that changed it into a metacentric (IIm). This metacentric IIm was present in a Hardy--Weinberg ratio throughout the collection range of more than 300 km. A number of aneuploids were found, pentasomics [2n – 1 = 23 (4.9%)], heptasomics [2n + 1 = 25 (2.5%)], and one octasomic [2n + 2 = 26 (0.3%)]. Re-assorted karyotypes with 2n = 24, but with odd numbers of two of the chromosomes, were also present (6.6%). B-chromosomes were found in root meristems and in pedicels in a number (19.1%) of the plants that grew around a 30 km marsh. Their numbers ranged from 1B to 8Bs per plant peaking in the 2B mode. The B-chromosomes (two polymorphs with terminal and near-terminal sticking points) were telocentric with a few metacentric iso-Bs. No B-carrying plants were found in the drier regions away from the marsh. The B-frequency distribution for aneuploids and pericentric inversions was much different than for the standard euploid. Dr Mark Gettner died on November 2nd, 2002. He is fondly remembered by his family, his many students and friends for his kindness and generosity, and for his dedication to his scientific work. Dr Broun is the son-in-law of Dr Gettner and he will be the corresponding author for this article (pb22@york.ac.uk).     

Karyotypic analysis of <Emphasis Type="Italic">Skimmia japonica</Emphasis> (Rutaceae) and related species

A karyotypic analysis of three species of Skimmia (Rutaceae) in East Asia was performed that examined 88 individuals from 53 localities. Chromosome numbers of S. japonica, S. reevesiana and S. arisanensis were 2n=30, 31, 32 (=30+0−2B), 2n=60 and 2n=60, respectively. The chromosome number of S. arisanensis was reported for the first time. All species had a large chromosome pair or quartet (the first pair or quartet) with a median–submedian centromere in the karyotype. In S. japonica the arm ratio of this first pair was considerably variable and showed a geographical pattern. In the northern half of the distribution range, Sakhalin, Hokkaido, Honshu, Shikoku and part of Kyushu, the arm ratio was 1–1.2, while in the southern half, part of Kyushu, Ryukyu and Taiwan, the arm ratio was very variable and ranged from 1.2 to 2.4. In S. japonica the first pair was sometimes rather heteromorphic; however, the heteromorphism was not related to sex of the plant.     

Elimination and duplication of particular Hordeum vulgare chromosomes in aneuploid interspecific Hordeum hybrids

Summary Seeds formed in crosses Hordeum lechleri (6x) x H. vulgare (2x and 4x), H. arizonicum (6x) x H. v. (2x), H. parodii (6x) x H. v. (2x), and H. tetraploidum (4x) x H. v. (2x) produced plants at high or rather high frequencies through embryo rescue. Giemsa C-banding patterns were used to analyze chromosomal constitutions and chromosomal locations on the methaphase plate. Among 100 plants obtained from H. vulgare (2x) crosses, 32 plants were aneuploid with 2n=29 (1), 28 (3), 27 (13), 26 (5), 25 (4), 24 (4), or 22 (2); 50 were euploid (12 analyzed), and 18 were polyhaploid (5 analyzed). Four plants had two sectors differing in chromosome number. Two of four hybrids with H. vulgare (4x) were euploid and two were aneuploid. Parental genomes were concentrically arranged with that of H. vulgare always found closest to the metaphase centre. Many plants showed a certain level of intraplant variation in chromosome numbers. Except for one H. vulgare (4x) hybrids, this variation was restricted to peripherally located non-H. vulgare genomes. This may reflect a less firm attachment of the chromosomes from these genomes to the spindle. Interplant variation in chromosome numbers was due to the permanent elimination or, far less common, duplication of the centrally located H. vulgare chromosomes in all 34 aneuploids, and in a few also to loss/gain of non-H, vulgare chromosomes. This selective elimination of chromosomes of the centrally located genome contrasts conditions found in diploid interspecific hybrids, which eliminate the peripherally located genome. The difference is attributed to changed genomic ratios. Derivatives of various H. vulgare lines were differently distributed among euploid hybrids, aneuploids, and polyhaploids. Chromosomal constitutions of hypoploid hybrids revealed a preferential elimination of H. vulgare chromosomes 1, 5, 6, and 7, but did not support the idea that H. vulgare chromosomes should be lost in a specific order. H. vulgare SAT-chromosomes 6 and 7 showed nucleolar dominance. Aneuploidy is ascribed to the same chromosome elimination mechanism that produces haploids in cross-combinations with H. vulgare (2x). The findings have implications for the utilization of interspecific Hordeum hybrids.     

“Cuckoo” Aegilops addition chromosome in wheat ensures its transmission by causing chromosome breaks in meiospores lacking it

In monosomic additions of Aegilops sharonensis to Chinese Spring wheat (2n=42 wheat chromosomes + 1 homoeologous group 4 Aegilops chromosome known as 4S¹), probably all functional gametes carried one Aegilops chromosome. Such preferential transmission is unusual in monosomic alien additions. Male and female meiosis seemed usually normal, but about 75% of embryo sacs and 28% of pollen grains were visibly abnormal near anthesis. Before the first gametophyte mitosis, up to about 13% of megaspores and pollen grains showed abnormalities usual in wheat aneuploids. However, in first mitosis, 50% of megaspores at metaphase and anaphase and 41% of pollen grains at anaphase and telophase contained acentric chromosome (or chromatid) segments of various sizes, up to about 32 in a cell, which were separated from the rest of the chromosome (or chromatid) by a gap or a thin Feulgen-positive thread. Such separated segments (SSs) are not normally seen in wheat and its aneuploids. The data indicat that alien and wheat chromosomes interact in meiocytes so that meiospores with the alien chromosome develop into normal gametophytes, but meiospores lacking the alien chromosome have SSs at first mitosis by whose loss or unequal distribution between daughter nuclei sterilizing deficiencies arise. Thus only gametophytes with the alien chromosome are competent.     

ANEUSOMATY IN ANEUPLOID POPULATIONS OF CLAYTONIA VIRGINICA

Lewis Walter H. (Stephen F. Austin State Coll., Nacogdoches, Texas.) Aneusomaty in aneuploid populations of Claytonia virginica. Amer. Jour. Bot. 49(9): 918–928. Illus. 1962.—From 2 central east Texas populations of Claytonia virginica, 15 chromosome numbers, 2n = 14, 15, 16, 18, 25, 26, 27, 28, 29 30, 31, 32, 33, 36, and 58, were found among a sample of 181 plants. The most frequently encountered numbers were 2n = 14, 28, and 29. Among an additional 14 plants the pollen mother cells in the same bud differed from one another in chromosome number, as well as the pollen and premeiotic cells from the same plant. The chromosomes of the most unstable plant varied from 2n = 14–36. Numerous meiotic abnormalities, including inversions, dicentrics, bridges, fragments, non-disjunctions, univalents, and multivalents, were observed for the aneusomatic and trisomic plants. It is suggested that the origin of the aneusomatics is related to the numerical disparity of the gametic chromosomes composing them. Since the species is perennial in habit, thereby allowing the unstable plants to produce gametes with varying chromosome numbers year after year, it is further proposed that the wide range of aneuploid known for C. virginica resulted, at least in part from the presence of aneusomatic individuals.     

BIOSYSTEMATIC STUDIES IN THE BOUTELOUA CURTIPENDULA COMPLEX. I. THE ANEUPLOID RHIZOMATOUS B. CURTIPENDULA OF TEXAS

Gould , F. W., and Z. J. Kapadia . (A. & M. College of Texas, College Station.) Biosystematic studies in the Bouteloua curtipendula complex. I. The aneuploid rhizomatous B. curtipendula of Texas. Amer. Jour. Bot. 49(8): 887–891. Illus. 1962.—Widespread throughout central U.S. is a rhizomatous form of B. curtipendula that basically is tetraploid (2n = 40). In the southwest the predominant type is a caespitose aneuploid with a high chromosome number (2n = ca. 80 to 2n = ca. 102). The present study has shown the presence of an extensive series of rhizomatous aneuploids in central Texas, with chromosome numbers ranging from 2n = 41 to 2n = 64. The distribution of these plants is centered about the region of overlap in the ranges of the 2 previously mentioned types. Available evidence indicates that the rhizomatous aneuploids have arisen through hybridization of the caespitose aneuploids and the rhizomatous tetraploids.     

Colonization history of the Japanese water shrewChimarrogale platycephala, in the Japanese Islands

To evaluate the intraspecific evolutionary history and local differentiation of the Japanese water shrewChimarrogale platycephala (Temminck, 1842), we an a lyzed the mitochondrial cytochromeb (Cytb) sequence divergence for samples from 55 localities in the Japanese is lands of Honshu and Kyushu. According to phylogenetic trees based on theCytb data, there were fourCytb haplotype lineages, which showed rough affinities with geographic areas, namely, Eastern/Central Honshu, the Kinki District of Western Honshu, the Chugoku District of Western Honshu, and Kyushu. However, in the alpine areas of the boundary between the Kinki and Chugoku Districts, complicated distribution patterns of theCytb haplotypes were revealed. Considering the present data and geological history in the Quaternary, we hypothesized the following evolutionary scenario. First, differ entiation and division into four primary ancestral geographic colonies of the shrews occurred in hypothetical refugia in the mid — late Pleistocene. Subsequently, rapid expansion occurred and caused the complicated distribution patterns of theCytb haplotypes in the boundary areas owing to the complex topography during the late stage of the Quaternary.     

Asynchronous meiotic rhythm as the cause of selective chromosome elimination in an interspecific <Emphasis Type="Italic">Brachiaria</Emphasis> hybrid

This paper reports the occurrence of chromosome elimination during microsporogenesis in an interspecific hybrid between a sexual diploid accession (SEX) of Brachiaria ruziziensis (2n=2x=18) and an apomictic tetraploid accession (APO) of B. brizantha (2n=4x=36). Meiosis was very abnormal in the triploid hybrid (2n=3x=27); we observed a distinct asynchrony from metaphase I to the end of meiosis. The APO and the SEX genomes did not show the same meiotic rhythm. When the former, with nine bivalents, was in metaphase I, the nine SEX univalents were not yet aligned; when the latter reached the plate, the APO genome was already in anaphase. In subsequent stages, the APO genome had reached the poles while the SEX was undergoing sister-chromatid segregation. As the SEX genome always remained temporally behind, it gave rise to one extra-nucleus in each pole. In the second division, the behavior was the same but anaphase II did not occur for the SEX genome, and only one extra-nucleus was observed in each cell in telophase II. Chromosome elimination for the SEX genome ranged from partial to total. The importance of these findings with respect to Brachiaria breeding programmes is discussed.     

Karyomorphological study of the Spiraea japonica complex (Rosaceae)

Reported in this paper is a karyomorphological study on one natural population of each of eight varieties in theSpiraea japonica complex. The interphase and mitotic prophase can be classified into the simple chromocenter type and the proximal type, respectively. The metaphase karyotypes of the eight varieties were formulated as follows:S. japonica var.japonica: 2n=18=14m+2sm+2st;S. japonica var.acuta: 2n=18=11m+4sm+3st;S. japonica var.incisa: 2n=18=12m+4sm+2st;S. japonica var.stellaris: 2n=18=15m+1sm+2st;S. japonica var.acuminata: 2n=18=14m+2sm+2st;S. japonica var.ovalifolia: 2n=18=10m+2sm+2st+4t;S. japonica var.glabra: 2n=18=10m+4sm+4st;S. japonica var.fortunei: 2n=36=17m+16sm+3st. Karyomorphological study reveals that the chromosome numbers within the complex are stable, and that karyomorphological divergence between the varieties lies mainly in chromosome size and organization. Based on the karyomorphological data and geographical distribution of the complex, the differentiation pattern as well as evolutionary mechanism of the complex is evaluated.     

<Emphasis Type="Italic">In vitro</Emphasis> root induction in weedy <Emphasis Type="Italic">amaranthus</Emphasis> species to obtain mitotic chromosomes

Summary Mitotic chromosome analysis has proven to be an important tool in monitoring the potential for genetic exchange among related plant species. One major obstacle to using mitotic chromosome analysis in any species is obtaining large numbers of clear, well-spread metaphase chromosomes necessary to perform cytological techniques such as chromosome banding and fluorescent in situ hybridization. The ability to obtain good chromosome spreads is in part determined by the number and morphology of the roots, which contain the metaphase tissue. Many Amaranthus species produce very thin, delicate roots. The technique used in the process described herein provides for much more substantial roots, allowing for higher probability of obtaining well-spread metaphase chromosomes. Seeds were planted in a soilless mixture, and then cuttings and leaves were taken from the plants. The cuttings were sterilized and placed in Murashige and Skoog (MS) media, while leaf tissue was analyzed by flow cytometry, both pre-and post-propagation, to obtain DNA contents. No changes in DNA content were observed. The in vitro procedure produced significantly larger roots than were produced in soilless mix. Furthermore, all of the in vitro roots observed had 32 chromosomes of normal morphology. In vitro root propagation allowed large numbers of roots to be obtained from a single plant, thereby resulting in increased probability of obtaining cells with metaphase chromosomes that reflected the original plants' chromosome numbers and therefore may be used for molecular cytogenetic analysis.     

RAD‐seq linkage mapping and patterns of segregation distortion in sedges: meiosis as a driver of karyotypic evolution in organisms with holocentric chromosomes

Meiotic drive, the class of meiotic mechanisms that drive unequal segregation of alleles among gametes, may be an important force in karyotype evolution. Its role in holocentric organisms, whose chromosomes lack localized centromeres, is poorly understood. We crossed two individuals of Carex scoparia (Cyperaceae) with different chromosome numbers (2n = 33^II = 66 × 2n = 32^II = 64) to obtain F1 individuals, which we then self‐pollinated to obtain second‐generation (F2) crosses. RAD‐seq was performed for 191 individuals (including the parents, five F1 individuals and 184 F2 individuals). Our F2 linkage map based on stringent editing of the RAD‐seq data set yielded 32 linkage groups. In the final map, 865 loci were located on a linkage map of 3966.99 cM (linkage groups ranged from 24.39 to 193.31 cM in length and contained 5–51 loci each). Three linkage groups exhibit more loci under segregation distortion than expected by chance; within linkage groups, loci exhibiting segregation distortion are clustered. This finding implicates meiotic drive in the segregation of chromosome variants, suggesting that selection of chromosome variants in meiosis may contribute to the establishment and fixation of chromosome variants in Carex, which is renowned for high chromosomal and species diversity. This is an important finding as previous studies demonstrate that chromosome divergence may play a key role in differentiation and speciation in Carex.     

Meiotic configuration and chromosome number in some Iranian species of Elymus L. and Agropyron Gaertner (Poaceae: Triticeae)

Chromosome numbers and analyses of meiotic metaphase I are reported for the following taxa: Agropyron cristatum subsp. incanum (2n= 42), A. cristatum subsp. pecttnatum (2n=28 – 33), Elymus elongatus subsp. ponticus (2n= 69, 70), E. hispidus var. hispidus (2n= 41 43), var. podperae (2n= 42) and var. villosus (2n= 41, 42), E. libanoticus (2n= 14), E. pertenuis (2n= 28, 28+1B), E. repens (2n= 42), E. transhyrcanus (2n= 40–42), E. hispidus var. villosus x E. cf. repens (2n= 42). Chromosome numbers only are reported for the following taxa: E. gentri (2n= 41, 42), E. nodosus subsp. dorudicus (2n= 28), and E. elongatiformis (2n= 56, 57). The haploid genomic constitution SP is reported for Elymus pertenuis. Variable chromosome numbers (2n= 28–32) were observed in the meiotic metaphase I within single anthers of Agropyron cristatum subsp. pectinatum, and the supernumerary chromosomes in this taxon are assumed to have originated from crosses with hexaploids. Partial elimination of these supernumerary chromosomes probably occurs during archesporial mitotic divisions or at an early stage in the meiotic cycle. A hybrid, morphologically intermediate between E. hispidus and E. repens, was obtained from a seed of E. hispidus collected in the field. The meiotic metaphase I configuration in this E. hispidus hybrid suggests that the pollen parent may itself be a hybrid or hybrid derivative of E. repens x E. hispidus.     

Karyomorphological studies onWoodwardia sensu lato of Japan

Karyomorphological comparisons were made of five species of JapaneseWoodwardia. There were no marked differences at interphase and prophase among the five species.Woodwardia japonica, W. prolifera, andW. unigemmata were diploid with 2n=68 and the formulas of their metaphase karyotypes uniformly 4m(median centromeric chromosomes)+12sm(submedian)+52(st+t)(subterminal and terminal).Woodwardia orientalis was tetraploid with 2n=136 and 8m+24sm+104(st+t), and the ratio of each chromosomal type to total complement was identical to that of three diploid species. These four species had several characteristics in common:x=34, the longest chromosome of sm, and a mean chromosome length over 3.0 μm. AlthoughWoodwardia orientalis showed some similarity toW. prolifera, it seems to be an allotetraploid which originated by chromosome doubling of a hybrid ofW. prolifera and a diploid species as yet karyomorphologically unknown.Woodwardia kempii was tetraploid with 2n=124 and 8m+24sm+92(st+t), and differed from the others in havingx=31, the longest chromosomes of t, and a mean chromosome length under 3.0 μm. This species has been classified as an independent genus,Chieniopteris, and our karyomorphological study supports this treatment.     

Characterization of Some Chromosomal Aberrations in Regenerated Rice Plants (<Emphasis Type="Italic">Oryza sativa</Emphasis>)

Somaclonal variation was studied in two Iranian land races of O. sativa spp. japonica var Hassani and O. sativa spp. indica var Sadridomsiah and 2000 plants of each cultivar were cytogenetically examined in two steps. In the first step, chromosome counts of root tips was used to detect ploidy levels and aneuploids of regenerated plants. In the second step, chromosomal aberrations were characterized by pachytene analysis of PMC’s. Ploidy levels were seen between n and 4n (haploids to tetraploids) in both cultivars with diploid resource (2n = 2x = 24). The total rate of variation for Hassani (japonica) was 13.7% including 10.8% for changes in chromosome number (ploidy levels and aneuploids) and 2.9% for chromosomal aberrations such as deficiency. A total rate of variation for Sadridomsiah (indica) was 15.6% including 12.4% for change in numbers and 3.2% for aberrations in construction. Most of important cytological mutations were observed in various chromosomes among regenerated plants of cultivars. Neither nullisomics nor inversions were distinguished in any samples.     

Caryological analysis of South American species of Vernonia (Vernonieae,Asteraceae)

Abstract

In the present study 16 populations belonging to 13 species of the genus Vernonia Schreb. were examined cytologically. In total, six different chromosome numbers, which represent three basic numbers: x = 10, x = 16 and x = 17, were found. These results include the first chromosome number reports for the following four species: V. lanifera Cristóbal & Dematt. (2n = 2x = 32), V. membranacea Gardner (2n = 2x = 34), V. salzmannii DC. (2n = 2x = 20) and V. scabrifoliata Hieron. (2n = 2x = 128). Besides, a new chromosome number was found in V. saltensis Hieron. (2n = 2x = 32), for which only tetraploid populations have been previously recorded. The data obtained in this work, along with the information available from the literature, show that the genus Vernonia in South America is heterogeneous with basic chromosome numbers that range between x = 9 and x = 19. These numbers suggest that a combination of polyploidy and aneuploidy has played an important role in the evolution of the genus.     

GISH analysis of disomic Brassica napus-Crambe abyssinica chromosome addition lines produced by microspore culture from monosomic addition lines

Two Brassica napus--Crambe abyssinica monosomic addition lines (2n=39, AACC plus a single chromosome from C. abyssinca) were obtained from the F₂ progeny of the asymmetric somatic hybrid. The alien chromosome from C. abyssinca in the addition line was clearly distinguished by genomic in situ hybridization (GISH). Twenty-seven microspore-derived plants from the addition lines were obtained. Fourteen seedlings were determined to be diploid plants (2n=38) arising from spontaneous chromosome doubling, while 13 seedlings were confirmed as haploid plants. Doubled haploid plants produced after treatment with colchicine and two disomic chromosome addition lines (2n=40, AACC plus a single pair of homologous chromosomes from C. abyssinca) could again be identified by GISH analysis. The lines are potentially useful for molecular genetic analysis of novel C. abyssinica genes or alleles contributing to traits relevant for oilseed rape (B. napus) breeding.