Assessment of genetic diversity among and within Carthamus species using sequence-related amplified polymorphism (SRAP) markers

Due to precise evaluation of genetic diversity of Carthamus species, sixty-two genotypes consisting fifty-two from five wild (C. oxyacanthus M. Bieb, C. lanatus L., C. dentatus Vahl, C. boissieri Halácsy, C. glaucus M.B.) and ten from cultivated species (C. tinctorius L.) were selected for evaluation of the genetic diversity in Carthamus species. A total of 238 (81.2 %) polymorphic bands were detected by 12 SRAP primer combinations with an average of 22 bands per combination. Me4-Em1 and Me5-Em2 primer combinations were known as the most informative SRAP markers based on the PIC values (0.34) where they distinguished all studied Carthamus species. Cluster analysis classified all accessions into five main groups among which clusters containing cultivated individuals were distinctly separated from those containing wilds. The most and the least genetic variation based on analysis of molecular variance, were detected within (76.90 %) and among (22.84 %) groups, respectively. The obtained results suggested that C. dentatus, C. glaucus and C. boissieri species may be classified in one section including C. dentatus in one and C. glaucus and C. boissieri in another subsection. The results also revealed high genetic similarity between C. oxyacanthus and C. tinctorius despite their different morphological characteristics.     

CYTOGENETICS OF HYBRIDS OF CARTHAMUS SPECIES (COMPOSITAE) WITH TEN PAIRS OF CHROMOSOMES

Plants thought to be typical of 7 species were chosen to represent the various taxa of Carthamus species with 10 pairs of chromosomes. These entities were crossed in all possible combinations and 20 of the possible 21 interspecific hybrids were obtained after 3 seasons of crosses. Analyses of the hybrids included studies of microsporocytes, pollen stainability, achene fertility, rudimentary ovaries, and other morphological characteristics. Pairing of chromosomes at metaphase I indicated no translocations were present in hybrids between C. tenuis from Israel, C. alexandrinus from Egypt, C. glaucus from northern Israel, and C. syriacus from Jordan. Members of this group are assigned the standard arrangement. Hybrids of C. glaucus from Iran, C. glaucus from Syria, and C. dentatus from Turkey always showed a translocation or chromosomal interchange when crossed with any member having the “standard” arrangement. The last 3 species are considered to have the “non-standard” chromosomal arrangement. The parental species used in this study can be regarded as a set of testers which will allow identification of chromosomal differentiation in additional Carthamus materials as they are collected.     

Discrimination and genetic diversity of cultivated and wild safflowers (Carthamus spp.) using EST-microsatellites markers

In spite of being one of the major oilseed crops, little is known about genetic diversity and relationships between species of safflower. In this study EST-SSR markers were used to evaluate and characterize 42 genotypes from six species including Carthamus tinctorius, Carthamus palaestinus, Carthamus oxyacanthus, Carthamus lanatus, Carthamus dentatus, and Carthamus boissieri. Thirty three primer pairs produced 123 polymorphic bands with 2–8 alleles per locus. The EST-SSR markers showed different level of gene diversity. The highest Polymorphism Information Content (PIC) values were observed for primers EL510507 and EL390720 (0.49 and 0.45, respectively). The highest genetic diversity and heterozygosity were observed for C. oxyacanthus. Both cluster and principal coordinate analysis (PCoA) clearly separated species into distinct groups. Within each species the accessions were clustered in different subgroups that mainly supported the known origins. The result showed that C. palaestinus had the most genetic similarity with cultivated safflower and C. oxyacanthus was next in this respect. In general, EST-SSR markers effectively revealed the genetic relationships and diversity of Carthamus species. This information is valuable for safflower improvement since C. palaestinus and C. oxyacanthus are both crossable with the cultivated species C. tinctorius.     

Species-genomic relationships among the tribasic diploid and polyploid Carthamus taxa based on physical mapping of active and inactive 18S–5.8S–26S and 5S ribosomal RNA gene families,and the two tandemly repeated DNA sequences

In the genus Carthamus (2n = 20, 22, 24, 44, 64; x = 10, 11, 12), most of the homologues within and between the chromosome complements are difficult to be identified. In the present work, we used fluorescent in situ hybridisation (FISH) to determine the chromosome distribution of the two rRNA gene families, and the two isolated repeated DNA sequences in the 14 Carthamus taxa. The distinctive variability in the distribution, number and signal intensity of hybridisation sites for 18S–26S and 5S rDNA loci could generally distinguish the 14 Carthamus taxa. Active 18S–26S rDNA sites were generally associated with NOR loci on the nucleolar chromosomes. The two A genome taxa, C. glaucus ssp. anatolicus and C. boissieri with 2n = 20, and the two botanical varieties of B genome C. tinctorius (2n = 24) had diagnostic FISH patterns. The present results support the origin of C. tinctorius from C. palaestinus. FISH patterns of C. arborescens vis-à-vis the other taxa indicate a clear division of Carthamus taxa into two distinct lineages. Comparative distribution and intensity pattern of 18S–26S rDNA sites could distinguish each of the tetraploid and hexaploid taxa. The present results indicate that C. boissieri (2n = 20) is one of the genome donors for C. lanatus and C. lanatus ssp. lanatus (2n = 44), and C. lanatus is one of the progenitors for the hexaploid (2n = 64) taxa. The association of pCtKpnI-2 repeated sequence with rRNA gene cluster (orphon) in 2–10 nucleolar and non-nucleolar chromosomes and the consistent occurrence of pCtKpnI-1 repeated sequence at the subtelomeric region in all the taxa analysed indicate some functional role of these sequences.     

DNA sequence diversity and the origin of cultivated safflower (<Emphasis Type="Italic">Carthamus tinctorius</Emphasis>L.; Asteraceae)

Background  

Safflower (Carthamus tinctorius L.) is a diploid oilseed crop whose origin is largely unknown. Safflower is widely believed to have been domesticated over 4,000 years ago somewhere in the Fertile Crescent. Previous hypotheses regarding the origin of safflower have focused primarily on two other species from sect. Carthamus – C. oxyacanthus and C. palaestinus – as the most likely progenitors, although some attention has been paid to a third species (C. persicus) as a possible candidate. Here, we describe the results of a phylogenetic analysis of the entire section using data from seven nuclear genes.     

5种裸子植物花粉外壁成分及结构分析比较

利用傅里叶变换红外光谱仪对银杏、日本冷杉等5种裸子植物的花粉外壁成分进行测定分析,结果表明,这5种裸子植物花粉外壁的红外光谱主要由蛋白质、脂类及多糖类物质的特征吸收峰组成。但不同科属间的花粉外壁主要成分含量存在较大差异,其中银杏科的银杏花粉外壁以蛋白质含量最为丰富;松科的雪松花粉外壁以脂类物质较为丰富;杉科的日本冷杉、杉木和日本柳杉花粉外壁成分以多糖类物质为主,但种间花粉外壁成分仍存在差异。采用扫描电镜观察5种裸子植物花粉,显示银杏、柳杉和杉木花粉粒体积较小,不具气囊,银杏花粉粒外壁表面具较均一条纹状纹饰,日本柳杉和杉木花粉粒外壁具颗粒状突起。雪松和日本冷杉花粉粒具气囊,体积较大,花粉外壁分别是粗糙具小穴状纹理以及表面光滑具微穿孔。     

Some evidence for infrageneric classification in Ziziphora L. (Lamiaceae:Mentheae)

The genus Ziziphora L. (Lamiaceae) consists of annual herbs, except Ziziphora clinopodioides, and is represented by six taxa in the flora of Turkey: Z. clinopodioides, Z. capitata, Z. persica, Z. tenuior, Z. taurica subsp. taurica and Z. taurica subsp. cleonioides. They are strong aromatic herbs and important medicinal plants. This study has been conducted to determine the palynological and karyological features of these taxa. The pollen morphology and exine structure of Ziziphora species were investigated by light microscope and scanning electron microscope. Pollen grains were determined as hexacolpate, medium in size and a circular or elliptic amb. The shape of pollen grains varies from oblate to prolate spheroidal. Ornamentation types of pollen grains are bireticulate, microreticulate or perforate. Somatic chromosome number has been determined and counted as 2n = 16, 18. Results indicate that the pollen characters and chromosome numbers of the genus are valuable for taxonomic applications and may be useful for infrageneric classification.     

Palynological investigations on some Hypericum taxa (Hypericaceae) growing naturally in Turkey

This study examined the detailed pollen morphological structures of sixteen Hypericum taxa (four endemic, twelve non endemic) including eight sections showing the natural distribution in Turkey: H. sect. Ascyreia (H. calycinum L.), H. sect. Heterophyllum (H. heterophyllum Vent.), H. Sect. Taeniocarpium (Hypericum confertum Choisy subsp. confertum, H. venustum Fenzl, H. linaroides Bosse), H. sect. Drosocarpium (H. montbretii Spach, H. bithynicum Boiss.), H. sect. Crossophyllum (H. adenotrichum Spach, H. orientale L.), H. sect. Olympia (H. olympicum L. subsp. olympicum), H. sect. Origanifolia (H. origanifolium Willd., H. avicularifolium Jaub and Spach subsp. depilatum (Freyn and Bornm.) Robson var. depilatum, H. avicularifolium Jaub. and Spach subsp. byzantinum) and H. sect. Hypericum (H. tetrapterum Fries, H. perforatum L., H. triquetrifolium Tura). These taxa were studied under light microscope and scanning electron microscope for the first time. Of the taxa studied, H. tetrapterum has the smallest pollen grains (on average 15.85–17.20 × 15.45–16.05 μm); and H. olympicum subsp. olympicum the largest grains (on average 22.90–23.10 × 21.40–22.10 μm). The pollen grains of 15 taxa are subprolate and one taxon is prolate-spheroidal. The regular pollen grains of all 16 taxa are 3-zonocolporate. Ornamentation is microreticulate; lumina regularly spaced in eight taxa, tectum perforatum; tectal perforations regularly spaced in seven taxa and tectum perforatum; and tectal perforations grouped together in one taxon. Endoaperture is cruciform porus, with short lateral and meridional extensions in ten taxa, cruciform porus, transversally elongated, with very small lateral extensions in one taxon and lalongate colpus in five taxa. Basic pollen types are ten taxa in type X, five taxa in type IV and one taxon in type II.     

A phylogenetic investigation of Carthamus combining sequence and microsatellite data

Carthamus (Asteraceae) includes both crop (Carthamus tinctorius, safflower) and weedy species. Despite previous studies, many outstanding questions remain regarding the phylogenetic surroundings of safflower, especially in relation to weedy species. Here we investigated relationships within Carthamus using a tiered approach examining sequence and microsatellite data. First, nuclear and chloroplast sequences were analyzed from 37 accessions of 16 species. Maximum parsimony, maximum likelihood, and Bayesian inference confirm two well supported clades, corresponding to sect. Atractlyis and sect. Carthamus, the latter of which includes safflower. Because sequence data provided limited resolution within the clades, microsatellite markers were used to investigate relationships within sect. Carthamus. Both sequence and microsatellite data reveal that most traditionally recognized species are not monophyletic. Microsatellite data indicate that Carthamus palaestinus is the closest relative of cultivated safflower.     

Cladistic and phylogenetic analyses of the genus Cicer in Turkey

Fifteen taxa of the genus Cicer L. growing naturally in Turkey and out-groups were classified by phylogenetic and cladistic analysis. Taxa of the genus Cicer and the out-group taxa belonging to the closest genera Phaseolus L., Vicia L., Lathyrus L. and Ononis L., which are placed in Phaseoleae, Vicieae and Ononideae tribes, respectively, were used in molecular studies in order to derive their phylogenetic relationships. Morphological, palynological and seed characters were used on the basis of 143 traits. The micromorphological characters of seed and pollen grains were revealed by SEM. Lathyrus L. and Vicia L. species were used as out-groups for numerical analysis. Ten specimens were used for the measurements of metric characters related to the morphological structures of the taxa used for statistical analysis via PAUP and NTSYS-pc packages. Phylogenetic relationships between species and populations of the same species growing in different locations and their variations were determined using molecular methods performed on regions of the Inter Simple Sequence Repeat (ISSR). DNA was isolated from the collected samples, using modified CTAB protocols. ISSR was used for phylogenetic fingerprinting. The data were analyzed with NTSYS-pc package. Standardized data were used to generate a dendrogram that revealed the phylogenetic relationships of the taxa. Geographic distribution of the Cicer taxa appears to be closely related to the Anatolian Diagonal. As a result of this study, four new endemic taxa were added and evaluated for the first time.     

Studies on the Pollen Morphology of Family Salicaceae in Gansu

Pullen grains of 2 genera 31 species of the family Salicaceae ia Gansu have been investigated. The comparative studies of the pollen grains were carried out by means of seanning electron microscope and light microscope. The two genera (Popu- lus L. and Salix L.) can be distinguished by their pollen characters such as the shape of pollen, type of aper ure, ornamentation of exine etc. 1. Shape of pollen grains 1) Spherical (rounded) : Populus L., 2) Ellipoidal, (subrotund): Salix L. 2. Type .of aperture 1) Atreme: Populus L., 2) 3-colpate, 3-colporoidate or 3-syncolpate: Salix L. 3. Ornamentation of exine 1 ) Cavernulous: Populus L., 2) Tubercular: Populus L., 3) Spinulate: Salix L., 4) Rugulose: Populus L. and Salix L., 5) Finely-reticulate: Populus L., 6) Reticulate: Salix L., 7) Crass-reticulate: Salix L., 8) Rugulose-Spinulate-Cavernulous: Salix L., 9) Rugulose-Reticulate: Salix L. Basing upon the above pollen characters, the genera Populus L. and Salix L. are con- sidered two good and natural taxa, and the genus Populus L. is more primitive than the genus Salix L.     

Insights into a hydration regulating system in Cupressus pollen grains

Background and Aims

Hydration, rupture and exine opening due to the sudden and large expansion of intine are typical of taxoid-type pollen grains. A hemispheric outgrowth external to the exine was observed on Cupressus and Juniperus pollen grains before the intine swelling and exine release. However, the actual existence of this permanent or temporary structure and its precise role in pollen hydration is still being debated. The aim of this paper is to collect information on the actual presence of this peculiar outgrowth on the surface of the Cupressus pollen grain, its structure, composition and function.

Methods

Pollen grains of several Cupressus species were observed using various techniques and methodologies, under light and fluorescence microscopy, phase-contrast microscopy, confocal microscopy, scanning electron microscopy, and an environmental scanning electron microscope. Observations were also performed on other species with taxoid-type pollen grains.

Key Results

A temporary structure located just above the pore was observed on Cupressus pollen grains, as well as on other taxoid-type pollens. It is hemispheric, layered, and consists of polysaccharides and proteins. The latter are confined to its inner part. Its presence seems to regulate the entrance of water into the grains at the beginning of pollen hydration.

Conclusions

The presence of a temporary structure over the pore of taxoid-type pollen grains was confirmed and its structure was resolved using several stains and observation techniques. This structure plays a role in the first phases of pollen hydration.     

Pollen morphology of some endemic Turkish Centaurea L. (Asteraceae, section Phaloletis) and their taxonomic value

In this study, the detailed pollen morphological structures of some endemic Turkish species of Centaurea amaena Boiss., C. antalyense H. Duman & A. Duran, C. aphrodisea Boiss., C. hierapolitana Boiss., C. luschaniana Heimerl, C. lycia Boiss., C. tossiensis Freyn. Et Sint., and C. wagenitzii Hub.-Mor. (Asteraceae, section Phaloletis) were studied under light microscopy (LM) and scanning electron microscopy (SEM) for the first time. LM and SEM investigations showed that the pollen grains of eight taxa are more or less spheroidal-subprolate, the amb triangular and tricolporate. The exine sculpture is tectate, microechinate-scabrate in the pollen of Centaurea taxa. Spinules are less dense in Centaurea amaena, C. antalyense, C. hierapolitana, and C. lycia, but they are more dense in C. aphrodisea, C. luschaniana, C. tossiensis, and C. wagenitzii. Spinule dimensions are different from each other. The exine has one layer of columellae beneath the spines. We determined all taxa that have the Helianthoid type. Exine sctructure and sculpture as well as spine density and dimensions in Asteraceae are the most reliable characteristics for discriminating taxa.     

八角属花粉形态研究

本文利用光学显微镜和扫描电镜对八角属(Illicium L.)14种1变种的花粉形态作了研究。通过花粉形态的研究,验证了在分类上本属所分各组的合理性,讨论了该属花粉形态的演化趋势。     

Pollen morphology of the genus Euphrasia L. (Orobanchaceae) from Pakistan and Kashmir and its taxonomic significance

Pollen morphology of 25 species of the genus Euphrasia L., belonging to the family Orobanchaceae from Pakistan has been examined by light and scanning electron microscope. It is a stenopalynous genus. Pollen grains are usually radially symmetrical, isopolar, tricolpate, sub-prolate to prolate-spheroidal rarely oblate-spheroidal. Sexine slightly thinner than nexine or as thick as nexine. Tectum coarse to fine retipilate or rugulate–retipilate. On the basis of exine ornamentation three distinct pollen types viz., Euphrasia foliosa, Euphrasia multiflora, and Euphrasia incisa are recognized. Pollen morphology is significantly helpful at specific level within the pollen type.     

刺五加、无梗五加花粉的亚显微形态学研究

用扫描电子显微镜观察了刺五加和无梗五加花粉的亚显微形态特征。花粉粒的赤道面观呈近圆形,极面观呈三裂圆形,具三孔沟。无梗五加、刺五加雄株的花粉表面纹饰为网状;刺五加两性株的花粉的表面纹饰亦呈网状,但有些花粉网脊不连续。无梗五加的花粉粒大小为26.5μm×27.4μm;刺五加雄株的花粉粒大小为21.1μm×22.1μm;刺五加两性株的花粉粒大小约为21.6μm×23.9μm。研究结果表明,无梗五加和刺五加雄株花粉的亚显微形态特征支持传统的属级和种级水平的分类;刺五加两性株某些花粉粒亚显微形态的独特表现可能与其发育不正常有关。     

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).     

Pollen morphology and anatomical features of Lilium (Liliaceae) taxa from Turkey

In this study, pollen grains and anatomical features of Turkish lilies were investigated under the electron (SEM) and light (LM) microscope. LM and SEM observations showed that the pollen grains are monosulcate, heteropolar, elliptical in polar view and oblate. Numerical results based on combined palynological and anatomical characters were discussed and compared with traditional taxonomic treatments. It was found that the midrib shape, mesophyll type, P/E (polar/equatorial), sulcus length, and lumina width are the most valuable traits in separating the examined taxa. The numerical analysis showed that Lilium candidum L. differs from the rest Turkish Lilium and also confirmed a close relationship between L. szovitsianum Fisch. & Avé-Lall. and L. armenum Miscz. ex Grossh. Also this study is the first report dealing with anatomical and palynological features of all Turkish lilies.     

Micromorphological observations on leaf and pollen of Capparis L. sect. Capparis (Capparaceae)

Sect. Capparis is represented by a single species, Capparis spinosa L., divided into several intraspecific taxa showing plesiomorphic features and disjunct distributions in the Old World. Leaf surface and pollen features were investigated in the whole group by SEM and light microscope observations. The section is characterized by simple hairs, a reticulate to undulate cuticle, anomocytic stomata surrounded by a peristomal rim, and trizonocolporate, prolate pollen grains. The characteristics of the indumentum appear constant, while the studied taxa are fairly differentiated with respect to cuticular patterns and dimensions of the stomata, and show slight differences in pollen size and exine surface. This micromorphological evidence, coupled with other phenotypic features, supports the placement of this section at the base of the genus Capparis in the paleotropical area. Considering the striking geographic disjunction and symplesiomorphies of the group, its biogeographical and systematic aspects are also discussed.