Karyological notes on another eight species of Achillea (Asteraceae) from Turkey

The chromosome number and morphology in eight species of the sections Ptarmica (Mill.) W. D. J. Koch, Anthemoideae (DC.) Heimerl, Arthrolepis Boiss., Santolinoideae (DC.) Heimerl and Achillea of the genus Achillea L. (Asteraceae) were investigated using karyological techniques. Sample plants and seeds of A. biserrata M. Bieb., A. fraasii var. troiana Aschers. & Heimerl, A. multifida (DC.) Boiss., A. brachyphylla Boiss. & Hausskn., A. pseudaleppica Hub.-Mor., A. cretica L., A. latiloba Ledeb. ex Nordm., and A. kotschyi Boiss. subsp. kotschyi) were collected from natural habitats in 2003 and 2004. The chromosome number found in seven species was 2n = 18, while only A. kotschyi had 2n = 36. All chromosomes had median point (M), median region (m), and submedian (sm) centromers. In addition, only A. biserrata species had one subterminal (st) chromosome. An increase in asymmetry was not observed in the karyotypes of the species studied. None of the studied species had any B chromosomes.     

Trinorguaian and germacradiene type sesquiterpenes along with flavonoids from the herbs of Pimpinella cappadocica Boiss. & Bal.

A trinorguaian-type sesquiterpenoid named kanalpin and two acylated flavonol glycosides named erzurumin and ilicanin have been isolated from Pimpinella cappadocica Boiss. & Bal., together with three known sesquiterpenoids and six known flavonoid glycosides. The structures of these compounds were determined by extensive spectroscopic (UV, IR, EI–MS, APCI–MS, HR-ESI-MS, 1D and 2D NMR) analyses. Antioxidant capacity of isolated secondary metabolites containing extracts of P. cappadocica was evaluated.     

Comparative karyotype analysis ofCeratozamia mexicana andMicrocycas calocoma (Zamiaceae) using fluorochrome banding (CMA/DAPI) and fluorescence in situ hybridization of ribosomal DNA

Orcein staining, differential staining with CMA and DAPI, and FISH with an rDNA probe were used to compare somatic chromosomes ofCeratozamia mexicana andMicrocycas calocoma. CMA-positive dots and hybridization signals appeared on chromosomes at early interphase and mitotic prophase, but in significantly different number in the two species. InCeratozamia mexicana, the CMA-positive and DAPI-negative bands and the hybridization signal were located at the terminal region of the long arm of three median-centromeric chromosomes, the terminal region of the short arm of two median-centromeric chromosomes and the terminal region of the long arm of two subterminal-centromeric chromosomes. InMicrocycas calocoma, they were located at the pericentric region of two median-centromeric chromosomes. These chromosome data suggested thatMicrocycas has no simple Robertsonian relationship toCeratozamia.     

THE HORDEUM VIOLACEUM COMPLEX OF IRAN

Fifty-seven Iranian collections of Hordeum violaceum Boiss. & Huet, a perennial forage grass, contained diploid (2n = 14), tetraploid (2n = 28), and hexaploid (2n = 42) chromosome races. All collections came from moderate to high elevations in the Alborz and Zagros mountains and adjacent plateau areas of Iran. Each chromosome race had a discrete distribution, and the hexaploids were the most widespread. The diploids were cytologically regular, except for a chromosome interchange that occurred in about half of the plants. The tetraploids and hexaploids behaved cytologically as autopolyploids. The hexaploids were taller, coarser and later-flowering than the diploids and tetraploids, and they had fewer but thicker culms and larger seeds. The tetraploids were the leafiest and most productive, making them the most desirable from an agronomic standpoint. All races were more or less self-sterile, a characteristic that sets H. violaceum apart from most other Hordeum species. The taxonomic status of H. violaceum and its closest relatives, H. turkestanicum Nevski and H. brevisubulatum Link, is uncertain because of close morphological similarities and the occurrence of chromosome races in each taxon.     

Reticulate evolution of the hybrid produced artificially by crosses between Osmunda banksiifolia and Osmunda lancea

Although ferns have been developed by hybridization and chromosome doubling, no natural polyploidy has yet been recorded in Osmundaceae. So, we produced hybrids artificially by crosses between Osmunda banksiifolia (2n = 2x = 44) and Osmunda lancea (2n = 2x = 44), and investigated their sporogenesis. From the O. banksiifolia × O. lancea hybrid with 44 univalent chromosomes, allotetraploids with 44 bivalent chromosomes were produced by chromosome doubling, and allotriploids with 22 univalent chromosomes and 22 bivalent chromosomes were then produced by back crosses. The results show when and how chromosome doubling occurs in hybrids. The success of artificial hybridization between O. banksiifolia and O. lancea, did not, however, reflect any product of natural hybridization between the two species.     

Karyomorphology and nuclear DNA content of sixteen Ophrys L. taxa from Turkey

Karyotypes in 16 representative taxa of the Ophrys genus are compared, based on Feulgen-stained somatic metaphase chromosomes. The karyotypes of O. omegaifera subsp. israelitica, O. ulupinara, O. lycia, O. argolica subsp. lucis, O. argolica subsp. lesbis, O. climacis and O. reinholdii subsp. reinholdii are described for the first time. Karyological analyses indicate relationships among the species with respect to their asymmetry indices. All Ophrys taxa studied were diploid with 2n = 2x = 36 chromosomes. One B chromosome has been detected among the chromosomes of O. argolica subsp. lucis. All karyotypes are symmetrical, consisting of metacentric and submetacentric chromosomes. The longest chromosomes of all the investigated specimens contain a secondary constriction. It is determined that there is a correlation between the total number of chromosomes having secondary constrictions and the evolutionary development order of the taxa. Based on nuclear DNA content, analysis was carried out by flow cytometer using propodium iodide as fluorochrome, 2C nuclear DNA content of 16 Ophrys species varying between 20.80 pg (O. argolica subsp. lucis) and 23.11 pg (O. omegaifera subsp. israelitica). Karyotype asymmetry relationships are discussed according to the bidimensional scatter plots of A₁–A₂, CV_CL–CV_CI, CV_CL–M_CA and CV_CI–M_CA.     

<Emphasis Type="Italic">Linaria khalkhalensis</Emphasis> (<Emphasis Type="Italic">Linaria</Emphasis> sect. <Emphasis Type="Italic">Linaria</Emphasis>, Scrophulariaceae), a new species from Iran

Summary   Linaria khalkhalensis Hamdi & Assadi is described as a new species from Azerbaijan and Guilan provinces (northern Iran), belonging to Linaria sect. Linaria. It is close to Linaria lineolata Boiss. and L. elymaitica (Boiss.) Kuprian., which it resembles in habit and vegetative features. However, floral and seed characters together with a distinctive geographical distribution, warrant recognition at the species rank. An illustration of the new species, a distribution map, scanning electron micrographs of seed and capsule surfaces, and an identification key are also included.     

Fissions,fusions, and translocations shaped the karyotype and multiple sex chromosome constitution of the northeast‐Asian wood white butterfly,Leptidea amurensis

Previous studies have shown a dynamic karyotype evolution and the presence of complex sex chromosome systems in three cryptic Leptidea species from the Western Palearctic. To further explore the chromosomal particularities of Leptidea butterflies, we examined the karyotype of an Eastern Palearctic species, Leptidea amurensis. We found a high number of chromosomes that differed between the sexes and slightly varied in females (i.e. 2n = 118–119 in females and 2n = 122 in males). The analysis of female meiotic chromosomes revealed multiple sex chromosomes with three W and six Z chromosomes. The curious sex chromosome constitution [i.e. W_1–3/Z_1–6 (females) and Z_1–6/Z_1–6 (males)] and the observed heterozygotes for a chromosomal fusion are together responsible for the sex‐specific and intraspecific variability in chromosome numbers. However, in contrast to the Western Palearctic Leptidea species, the single chromosomal fusion and static distribution of cytogenetic markers (18S rDNA and H3 histone genes) suggest that the karyotype of L. amurensis is stable. The data obtained for four Leptidea species suggest that the multiple sex chromosome system, although different among species, is a common feature of the genus Leptidea. Furthermore, inter‐ and intraspecific variations in chromosome numbers and the complex meiotic pairing of these multiple sex chromosomes indicate the role of chromosomal fissions, fusions, and translocations in the karyotype evolution of Leptidea butterflies.     

Cytogenetic effects of nine <Emphasis Type="Italic">Helichrysum</Emphasis> taxa in human lymphocytes culture

Helichrysum Mill. (Asteraceae) species have been used in folk medicine for thousands of years in the world. The in vitro cytogenetic effects in human lymphocytes of nine Helichrysum taxa used in Turkey folk medicine were investigated. Blood samples were obtained from healthy donors, non-smoking volunteers, which were incubated and exposed to increasing concentrations of methanol extracts of Helichrysum taxa (0.01, 0.05, 0.1, 0.5 and 1 mg/mL). The inhibitory effects of H. stoechas (L.) Moench subsp. barrelieri (Ten.) Nyman, H. armenium DC. subsp. armenium, H. armenium DC. subsp. araxinum (Kirp.) Takht., H. plicatum DC. subsp. plicatum, H. compactum Boiss. and H. artvinense P.H.Davis & Kupicha on the mitotic index and replication index indicate that these taxa can have genotoxic and mutagenic effects. They should therefore not be used freely in alternative medicine although their antiproliferative activity may suggest anticarcinogenic properties. Increase effects of H. stoechas subsp. barrelieri, H. armenium subsp. armenium, H. armenium subsp. araxinum, H. chasmolycicum P.H.Davis, H. plicatum subsp. plicatum, H. compactum and H. artvinense on the micronucleus rates showed that these taxa can have genotoxic and carcinogenic effects.     

Molecular cytogenetic characterization of <Emphasis Type="Italic">Rumex papillaris</Emphasis>, a dioecious plant with an XX/XY<Subscript>1</Subscript>Y<Subscript>2</Subscript> sex chromosome system

Rumex papillaris Boiss, & Reut., an Iberian endemic, belongs to the section Acetosa of the genus Rumex whose main representative is R. acetosa L., a species intensively studied in relation to sex-chromosome evolution. Here, we characterize cytogenetically the chromosomal complement of R. papillaris in an effort to enhance future comparative genomic approaches and to better our understanding of sex chromosome structure in plants. Rumex papillaris, as is common in this group, is a dioecious species characterized by the presence of a multiple sex chromosome system (with females 2n = 12 + XX and males 2n = 12 + XY₁Y₂). Except for the X chromosome both Y chromosomes are the longest in the karyotype and appear heterochromatic due to the accumulation of at least two satellite DNA families, RAE180 and RAYSI. Each chromosome of pair VI has an additional major heterochromatin block at the distal region of the short arm. These supernumerary heterochromatic blocks are occupied by RAE730 satellite DNA family. The Y-related RAE180 family is also present in an additional minor autosomal locus. Our comparative study of the chromosomal organization of the different satellite-DNA sequences in XX/XY and XX/XY₁Y₂ Rumex species demonstrates that of active mechanisms of heterochromatin amplification occurred and were accompanied by chromosomal rearrangements giving rise to the multiple XX/XY₁Y₂ chromosome systems observed in Rumex. Additionally, Y₁ and Y₂ chromosomes have undergone further rearrangements leading to differential patterns of Y-heterochromatin distribution between Rumex species with multiple sex chromosome systems.     

Centaurea rahiminejadii sp. nov. and a new record in the genus Centaurea (Asteraceae,Cardueae–Centaureinae) from Iran

A new species, Centaurea rahiminejadii Negaresh (Asteraceae) in C. sect. Cynaroides Boiss. ex Walp., is described and illustrated from the Kermanshah Province, west Iran. It is morphologically similar to Centaurea regia Boiss. subsp. regia. Diagnostic morphological of this and closely related taxa are discussed and presented. The ecology, habitat and conservation status of the new species are also described. In addition, Centaurea regia subsp. cynarocephala (Wagenitz) Wagenitz is reported as a new record to the flora of Iran. Finally, the geographic distribution of the new species and closely related species is presented and mapped.     

Genetic variability for esterase enzyme in Onobrychis species

Summary Understanding polymorphism at the enzyme level is basic to its use in population and genetic studies. However, no such information is available on the variability among different sainfoin (Onobrychis) species. Therefore, our objective was to study the existence of genetic polymorphism for esterase in 17 Onobrychis species and three cultivars of O. viciifolia Scop. Three regions of banding were observed in all the materials tested, with the number of bands varying from 0 to 3, 3 to 14, and 1 to 2 bands in each of these zones, which have been designated EST1, EST2, and EST3 respectively. All the materials studied had unique banding patterns, the only common feature being that all of them, except one species, had isozyme 1. Identification was possible only for four species (O. iberica, O. kachetica, O. transcaucasica, and O. bieberstenii) and one cultivar (Nova) based on the banding patterns. Large diversity was evident from the wide range of percent similarity values (0%–79%). Subsequent studies should be directed in using these isozyme banding patterns as markers to the desirable agronomic and quality traits of different germplasm lines.This work was supported by USDA Specific Cooperative Agreement No. 58-7MN1-8-143 from the Plant Stress and Water Conservation Unit, USDA-ARS, Lubbock, Texas. Joint contribution of the Texas Tech University, Lubbock, Texas and the USDA-ARS. TTU Journal no. T-4-302     

A monograph of <Emphasis Type="Italic">Octoknema</Emphasis> (Octoknemaceae — Olacaceae <Emphasis Type="Italic">s.l.</Emphasis>)

A revision of Octoknema Pierre is provided, based on morphological data gathered from a study of herbarium specimens and observations in the field. Fourteen species of Octoknema are recognised including six new species: O. bakossiensis Gosline & Malécot, O. belingensis Gosline & Malécot, O. chailluensis Malécot & Gosline, O. kivuensis Gosline & Malécot, O. mokoko Gosline & Malécot and O. ogoouensis Malécot & Gosline. Data are given for four additional poorly known taxa (Octoknema species A, B, C and D).     

Cytogenetic characterization of <Emphasis Type="Italic">Hydrangea involucrata</Emphasis> Sieb. and <Emphasis Type="Italic">H. aspera</Emphasis> D. Don complex (Hydrangeaceae)<Emphasis Type="Italic">:</Emphasis> genetic,evolutional, and taxonomic implications

The subsection Asperae of genus Hydrangea L. (Hydrangeaceae) has been investigated for three reasons: several ambiguous classifications concerning Hydrangea aspera have been published, unexpected differences in genome size among seven accessions have been reported Cerbah et al. (Theor Appl Genet 103:45–51, 2001), and two atypical chromosome numbers (2n = 30 for Hydrangea involucrata and 2n = 34 for H. aspera) have been found when all other species of the genus present 2n = 36. Therefore, these two species and four subspecies of Hydrangea in all 29 accessions were analyzed for their genome size, chromosome number, and karyotype features. This investigation includes flow cytometric measurements of nuclear DNA content and bases composition (GC%), fluorochrome banding for detection of GC- and AT-rich DNA regions, and fluorescent in situ hybridisation (FISH) for chromosome mapping of 5 S and 18 S-5.8 S-26 S rDNA genes. In the H. aspera complex, the genome size ranged from 2.98 (subsp. sargentiana) to 4.67 pg/2C (subsp. aspera), an exceptional intraspecific variation of 1.57-fold. The mean base composition was 40.5% GC. Our report establishes the first karyotype for the species H. involucrata, and for the subspecies of H. aspera which indeed present different formulae, offering an element of discrimination. FISH and fluorochrome banding revealed the important differentiation between these two species (H. involucrata and H. aspera) and among four subspecies of the H. aspera complex. Our results are in agreement with the Chinese classification that places the groups Kawakami and Villosa as two different species: Hydrangea villosa Rehder and Hydrangea kawakami Hayata. This knowledge can contribute to effective germplasm management and horticultural use.     

Comparative pollen morphology of sect. <Emphasis Type="Italic">Quinqueloculares</Emphasis> (Campanulaceae) in Turkey

The pollen morphology of Turkish species belonging to sect. Quinqueloculares (Boiss.) Phitos (Campanulaceae) were investigated with a light microscope (LM) and scanning electron microscope (SEM). All examined species are endemic, except Campanula crispa Lam. According to the results of this study, pollen grains of the examined taxa were triporate and spheroidal. Their sculpture was spinose and baculate (C. crispa Lam.); spinulose and microperforate (C. tomentosa Lam.); microperforate (C. vardariana Bocquet) or more commonly, microechinate (C. iconia Phitos, C. lyrata Lam. subsp. lyrata, C. hagielia Boiss., C. sorgerae Phitos, C. betonicifolia SM., C. karadjana Bocquet, C. telmessi Hub.-Mor. & Phitos, C. davisii Turrill).     

The in vivo anthelmintic efficacy of some Verbascum species growing in Turkey

Verbascum L. (Scrophulariaceae) species are used for desiccating wounds and as a fish poison in Anatolia as well as for diarrhea and dysentery of animals in several countries. To further evaluate their activity, methanolic extracts obtained from 13 Verbascum species growing in Turkey, including V. chionophyllum Hub.-Mor., V. cilicicum Boiss., V. dudleyanum (Hub.-Mor.) Hub.-Mor., V. lasianthum Boiss., V. latisepalum Hub.-Mor., V. mucronatum Lam., V. olympicum Boiss., V. pterocalycinum var. mutense Hub.-Mor., V. pycnostachyum Boiss. & Heldr., V. salviifolium Boiss., V. splendidum Boiss., V. stachydifolium Boiss. & Heldr. and V. uschackense (Murb.) Hub.-Mor. were evaluated for their in vivo anthelmintic activity. The extracts from V. lasianthum, V. latisepalum, V. mucronatum and V. salviifolum showed the highest inhibitory rates against Aspiculuris tetraptera at 100 mg/kg in mice. Additionally, extracts from V. dudleyanum and V. pterocalycinum var. mutense were found generally highly effective. The remaining species did not show any activity. Results of the present study support the utilization of these plant species employed in Turkish folk medicine.     

Evaluation of seed and leaf epidermis characters in the taxonomy of some annual species of the genus Papaver (Papaveraceae)

To get a better understanding of their taxonomic importance, we examined morphological, leaf epidermal and seed micromorphological characters of 19 taxa of annual Papaver L. belonging to the sections Papaver, Carinatae Fedde and Rhoeadium Spach. Among the studied taxa, the pattern of the coat of the seed testa cells and seed epidermal cell showed large variation. The granulate type of testa sculpturing was most common, but other types were represented by one or two species. In contrast, most of the taxa have broadly similar leaf epidermal characteristics. We conclude that variation in seed characters are significant for the taxonomy at the specific and generic levels, whereas leaf epidermis morphology, with few exceptions, is mostly only useful for separation of taxa within species. Based on the results, P. lacerum Popov is regarded as a synonym of P. commutatum Fisch. & C. A. Mey, while P. piptostigma Bienert ex Fedde, P. halophilum (Fedde) Cullen and P. tubuliferum Fedde are regarded as synonyms of P. macrostomum Boiss. et Huet ex Boiss. var. macrostomum. Further, Papaver gaubae Cullen & Rech. f. and P. bipinnatum C. A. Mey. are included into the synonymy list of P. glaucum Boiss. et Hausskn. and P. arenarium M.‐Bieb., respectively. In contrast, Papaver chelidoniifolium Fisch. & C. A. Mey. var. pinnatum Tavakkoli & Assadi from Gilan province is described as a new variety, and P. rhoeas L. var. strigosum Boenn. and var. glabrum Fedde and P. commutatum subsp. euxinum Kadereit are introduced as new records to Iran. The evolutionary and phylogenetic implications of the observed variation in seed characteristics are also discussed.