Crossability analysis in subgenus Tulipa of the genus Tulipa L.

A range of almost 1400 cross combinations has been carried out between 31 species of Tulipa subgenus Tulipa. Most of the sections are well separated reproductively from each other. The designated species aggregates in section Clusianae appeared to be intersterile. Also in section Kolpakowskianae the species, as far as used, cannot be combined. Some crosses between species of section Tulipanum succeeded. The species of sections Eichleres and Tulipa (including the cultivated tulip T gesneriana ) could be combined in various ways, although the highest level of seed set was achieved within either section. The crossability data support the current taxonomic division. The level of reproductive isolation between sympatric species is discussed, and the importance of crossability studies for plant breeding in Tulipa pointed out briefly.     

Species relationships and taxonomy inTulipa subg.Tulipa (Liliaceae)

Principal component and canonical variate analyses were used to analyse the variation of 34 morphological characters measured in 98 accessions ofTulipa subg.Tulipa, formerly known as sect.Leiostemones. In addition 43 accessions were analysed cytogenetically by means of C-banding methods. Based on the encountered variation and on geographical distribution data 30 species in five sections were recognized, of which the synonymy is stated. Seven new series in two different sections have been designated.     

Interspecific crosses between Tulipa gesneriana cultivars and wild Tulipa species: a survey

Summary Interspecific crosses were made between 76 Tulipa gesneriana cultivars and 28 botanical Tulipa species. Tulipa gesneriana proved fully congruent (all cross-combinations giving hybrid progeny) with members of the subsection Gesnerianae, was moderately affiliated (50% of all cross-combinations giving hybrid progeny) with representatives of the subsection Eichleres, and was found to be incongruent with most other Tulipa species (except T. stapfii of the subsection Oculussolis).Relationships between T. gesneriana and some representatives of the subsection Eichleres were addressed in more detail. Seeds were harvested in all cross-combinations, but the number of seeds depended on the T. gesneriana cultivar used as the female partner. Apparently, the maternal genotype did affect pre- or post-fertilization processes. T. gesneriana was not crossable with T. kaufmanniana of the subsection Spiranthera. In this specific case, however, a bridge cross was used to overcome incongruity. For this, T. kaufmanniana was first combined with T. greigii, and the hybrid was subsequently crossed with T. gesneriana.These results demonstrate possibilities for interspecific cross-combinations in tulips and have offered opportunities for the introduction of desired new traits in the cultivated tulip.     

Molecular analysis of species of Tulipa L. from Iran based on ISSR markers

Molecular characterization of Tulipa L. species can elucidate the relationships among the species and provide more information about the taxonomy of this valuable genus. In this study, the genetic relationship among 39 Tulipa accessions from Khorassan and Yazd Provinces, located in east and northeast Iran, were analyzed using inter-simple sequence repeat (ISSR) primers. Ten selected ISSR primers from 20 screened primers generated a total of 97 polymorphic DNA bands. Unweighted pair-group method of cluster analysis based on Dice similarity values separated the accessions into nine groups. Seven species were recognized within these groups, and T.?micheliana Hoog was the most frequently encountered species. The subgroups formed within both T.?micheliana and T.?lehmanniana Merckl. revealed a low level of diversity within these species. T.?biebersteiniana Schultes & Schultes fil. and T.?biflora Pallas accessions made a separate clusters. The grouping of accessions was generally consistent with principal coordinate analysis (PCA) and clearly showed the position of species in the subgenera and sections of Tulipa. These results clearly showed the usefulness of DNA fingerprinting for identification of Tulipa accessions, and it is imperative to collect and characterize more genetic variability from the other distribution areas of this genus.     

Molecular phylogenetic analysis of Tulipa (Liliaceae) based on noncoding plastid and nuclear DNA sequences with an emphasis on Turkey

We investigated the phylogenetic relationships in Tulipa in Turkey using DNA sequences from the plastid trnL‐trnF region and the internal transcribed spacer (ITS) of nuclear ribosomal DNA. We generated trnL‐trnF and nuclear ITS sequences for 11 Tulipa spp. from Turkey and compared the utility of trnL‐trnF and ITS sequences for phylogenetic analysis. Neighbor‐joining, Bayesian and maximum parsimony methods were implemented using the same matrices. Our study of Tulipa based on molecular data revealed congruent results with previous studies. Despite the relatively lower resolution of trnL‐trnF than that of ITS, both sequence matrices generated similar results. Three clades were clearly distinguished, corresponding to subgenera Tulipa, Eriostemones and Orithyia. It is not fully resolved whether Clusianae should be recognized as a separate section of subgenus Tulipa or a distinct subgenus. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 172 , 270–279.     

Reassessment of diversity and analysis of distribution in Tulipa (Liliaceae) in Uzbekistan

The diversity and distribution of the genus Tulipa in Uzbekistan are discussed; 34 taxa of Tulipa are recognized in Uzbekistan. The taxa were mapped using GIS software and their distributions were analyzed. Six species are endemic to the country. A national check‐list of Tulipa was created based on the classification developed by Zonneveld (2009). ‘Hot spot’ areas of tulip diversity in Uzbekistan are western Tien‐Shan (18 taxa), the western Pamir–Alay mountains (18 taxa) and the Turan lowland (5 taxa). An assessment of rare tulip species according to IUCN red list categories and criteria was performed for the first time for Uzbekistan. Evaluation of the latest classification of Tulipa clearly shows that further studies are required to arrive at a natural classification of the genus.     

Species relationships in the genus Tulipa

The relationships of diploid and polyploid species belonging to the sub-section Eriostemones of the genus Tulipa have been investigated from the point of view of chromosome morphology, relative DNA values, and meiotic pairing properties. Karyotype morphology is basically the same for all species and eight principal chromosome classes can be distinguished. The range of DNA values obtained by Feulgen cytophotometry for the diploid species is relatively small and provides little assistance in ascertaining the pattern of ploidy involved in the evolution of the group. Partial asynapsis and reverse loop pairing in several of the diploids at pachytene reveals a degree of structural differentiation which may be the outcome of hybridity between species. Significantly, these anomalies are rarely encountered at the equivalent tetraploid levels and a scheme is proposed to account for this behaviour.     

Effect of a gap on gene flow between otherwise adjacent transgenic <Emphasis Type="Italic">Brassica napus</Emphasis> crops

The chloroplast DNA diversity of 33 accessions belonging to 16 species of five sections in Allium subgenus Rhizirideum was studied by analysing the sequence of three fragments: the trnL-F intergenic spacer, the rps 16 intron and rbcL (rubisco large subunit). The three sections Cepa, Schoenoprasum and Rhizirideum, representing the majority of the included species, each possess a separate clade after phylogenetic analysis. Exceptions to this general rule are the placement of Allium pskemense (section Cepa) connected to Allium senescens (section Rhizirideum) and Alium roylei, taking an intermediate position between sections Cepa and Schoenoprasum. Both species were located in their own section after nuclear DNA analysis. A range of crossing experiments has been carried out. The different position of A. roylei when comparing cpDNA and nDNA diversity was not confirmed with the production of hybrid seeds after crossing A. roylei with species other than those of section Cepa. The different position of A. pskemense in the cpDNA and the nDNA tree can not be compared to its crossability, since only a few crossing experiments are reported for this species. The hypothesis that a shorter distance between two species in a cpDNA tree compared to their distance in a nDNA tree will indicate interfertility at a certain level, is neither confirmed nor rejected by the currently available results.Communicated by R. Hagemann     

Biosystematic studies inTulipa sect.Eriostemones (Liliaceae)

Evolutionary relationships and infra- and interspecific variability within the species ofTulipa sect.Eriostemones Boissier were studied. Measurements of 35 morphological characters were used for principal component and canonical variate analyses. Crossing experiments and chromosome counts were carried out. Two axes of morphological variation were encountered within the principal component plot. Variation of subsect.Biflores was traced along one axis, the species of subsect.Australes as well asSaxatiles were found along the other axis. The nomenclature of names of subsect.Biflores was considered on the basis of their geography, specific morphological characteristics and polyploidy. Conspecificity of names of all species was tested based on both morphology and crossability.     

Tetraploid wheat crossability with rye (Secale)

Hybridisations carried out between a number of tetraploid wheats (Triticum turgidum (L) Thell, ssp. turgidum conv. durum (Desf.) MacKey, N. comb.) and rye (Secale) species revealed a range from high to low levels of crossability between them.Variation in crossability level was due to genetic variation, both in tetraploid wheat and rye. The presence of a significant (p<0.01) interaction in crossability between tetraploid wheat and rye indicates that the genetic system governing crossability is more complex than previously described.It appears that the mutation(s) in wheat from high to low crossability with rye took place at least as early as the tetraploid level of wheat evolution.     

GENOME AND KARYOTYPE RELATIONSHIPS IN THE GENUS DENDROBIUM (ORCHIDACEAE). I. CROSSABILITY

A hybridization study was made utilizing 37 species of 10 sections in 41 intra- and intersectional combinations. A total of 721 pollinations was made with 138 fruits harvested; 89 produced viable crosses. Five intrasectional and 20 intersectional combinations resulted in successful hybrids. Among the intrasectional crosses, Phalaenanthe × Phalaenanthe exhibited the highest percentage of fruits and successful crosses while Callista × Callista produced no successful crosses. Eugenanthe × Eugenanthe exhibited low crossability. Among the intersectional crosses, Ceratobium × Phalaenanthe gave the highest percentage of successful crosses, indicating a close relationship between the two groups. Also, there appears to be a fairly close relationship between Latourea and Ceratobium based on crossability. Although the sectional classification of the genus by Schlechter on the basis of vegetative and floral morphology appears to be valid, crossability studies suggest that the Ceratobium and Phalaenanthe sections might be integrated and the Eugenanthe section separated into sections or subsections.     

Anthocyanin content of Tulipa species and cultivars and its impact on tepal colours

Flowers of tulips (17 species and 25 cultivars) were subjected to qualitative and relative quantitative examination for anthocyanins. Altogether five anthocyanins were identified as the 3-O-(6″-O-α-rhamnopyranosyl-β-glucopyranoside) of delphinidin (1), cyanidin (2) and pelargonidin (3), and the 3-O-[6″-O-(2‴-O-acetyl-α-rhamnopyranosyl)-β-glucopyranoside] of cyanidin (4) and pelargonidin (5). The pigments 1–5 represented 7%, 43%, 12%, 2% and 31%, respectively, of the total anthocyanin amount in the tepals of the Tulipa species, and 20%, 37%, 30%, 6% and 4%, respectively, in the cultivar tepals. Nearly 50% of the samples contained acetylated anthocyanins. The colours of the freeze-dried tepals described by the CIELab coordinates, hue angle (h_ab), saturation (C*), and lightness (L*) together with the anthocyanin content were subjected to multivariate analysis. All tepals classified with hue angles described as “blue nuances” were from cultivars. They contained 1 as the major anthocyanin, and no or just traces of pelargonidin derivatives. The species and cultivars having “magenta nuances” showed similar anthocyanin content with increased relative proportions of 2 at the expense of 1. Orange coloured tepals were to a large extent correlated with high relative proportions of the pelargonidin derivatives, 3 and 5. Acetylation of anthocyanins furnished a weak colour effect opposite to the bluing effect previously reported for anthocyanins with aromatic acyl groups. All six species belonging to the section Eichleres (subgenus Tulipa) were after principal component analysis grouped closely together. They were characterized by high concentrations of the pelargonidin derivatives 3 and 5, and orange petal nuances. However, within section Tulipa (subgenus Tulipa), considerable anthocyanin variation was observed. Species in the subgenus Eriostemones were generally characterized by the two anthocyanins 1 and 2, and no pelargonidin derivatives.     

Studies on sporopollenin biosynthesis in Tulipa anthers

Summary Extensive tracer experiments were carried out on Tulipa with the aim of determining the structure and biosynthesis of sporopollenin. The radiolabeled precursors were applied using an improved technique previously selected. The sporopollenin fraction was purified using either a gentle method — hydrolyzing enzymes (pronase, amylase, amyloglucosidase, cellulase, pectinase and lipase) and alkaline hydrolysis (method A) — or by a conventional aggressive procedure, where the material was enriched by alkaline hydrolysis and treated several days with 80% phosphoric acid (method B). The ¹⁴C-labeled precursors applied were mevalonate, glucose, acetate, malonic acid, phenylalanine, tyrosine, p-coumaric acid. Regardless of the method of enrichment, a higher level of incorporation into the sporopollenin fraction was always seen with [U-¹⁴C]-phenylalanine. The level of radioactivity found in sporopollenin labeled by phenylalanine or malonate was sufficiently high for the labeled polymer to be degraded and the products released analyzed for the first time. In the case of phenylalanine-labeled sporopollenin, the main degradation component, p-hydroxybenzoic acid, was also the most heavily labeled substance. This result was not dependent on the procedure used for sporopollenin enrichment. These findings are interpreted as meaning that phenylpropane metabolism via phenylalanine-ammonia lyase is involved in sporopollenin biosynthesis.     

Die Feinstruktur der Chromoplasten in plasmochromen Perigon-Blättern von Tulipa

Summary The chromoplasts of yellow perigon leaves of Tulipa derive from the young chloroplasts present in the bud stage. During chromoplast development and thylakoid breakdown many large plastoglobuli (diameter 80–360 nm) are formed. The function of the petal plastoglobuli as stores for the chromoplast lipids such as plastidquinones and carotenoids is discussed.     

The behavior of shoot apices of Tulipa in relation to floral induction

This study was carried out in three parts. Part I described the shoot apices of Tulipa as they are affected by temperature treatments which induce flowering. Part II traced the changes in the protein complement of the growing points that occurred concomitantly with floral induction. Part III dealt with several fractions of basic proteins with amino acid compositions that were consistent with their histone character. A general discussion deals with the overall problems of flowering in Tulipa.     

A new lectin from tulip (Tulipa) bulbs

A lectin was isolated from tulip (Tulipa) bulbs by affinity chromatography on fetuin-agarose and partially characterized. The tulip lectin is a tetrameric protein composed of four identical subunits of M_r 28 000, which are not held together by disulphide bonds. It is not glycosylated and has an amino-acid composition typified by a high content of asparagine-aspartic acid, leucine, glycine and serine. Tulip lectin agglutinates human red blood cells, but has a much higher specific activity with rabbit erythrocytes. In hapten-inhibition assays with the latter type of red blood cell the lectin exhibits a complex specificity, whereas its agglutination with human erythrocytes is readily inhibited by N-acetylgalactosamine, lactose, fucose and galactose.Abbreviations DEAE diethylaminoethyl - PBS phosphate-buffered saline - TL Tulipa lectin - M_r relative molecular mass - SDS-PAGE sodium dodecyl sulphate-polyacrylamide gel electrophoresis     

Species identification of the medicinal plant Tulipa edulis (Liliaceae) by DNA barcode marker

Tulipa edulis (Liliaceae) is the botanical origin of the traditional Chinese medicine (TCM) “Guangcigu”. Due to overexploitation that induced a decline in natural sources, many dried bulbs from other species of Tulipa have been used, adulterating the medicine in recent years. This practice may cause a series of inconsistent therapeutic effects and quality control problems in the herbal medicine industry. Hence, three DNA regions (matK, psbA-trnH and rbcL) were evaluated as barcodes for identifying T. edulis and its adulterants. All candidate DNA barcodes were successfully amplified from leaf samples. Based on the sequence divergences, rbcL and psbA-trnH can assign T. edulis and its adulterants to the correct genus, while matK can accurately differentiate T. edulis and its adulterants. Thus, at the DNA level, the matK intergenic region is a more suitable, accurate and applicable identification of T. edulis and its adulterants than rbcL and psbA-trnH.     

Inheritance of mucilage canals in Laminaria (section Simplices) in eastern Canada

Two experiments have been carried out to test the suitability of the occurrence of mucilage canals as a criterion for species or ecotype definition in the non-digitate section of the genus Laminaria. A matrix of crossability tests shows complete interfertility between all mucilage canal types, fertile F₁ hybrids being produced in all cases. Quantitative genetic analysis reveals a large environmental component in the phenotypic variance of degree of mucilage canal development. Only plants from Nova Scotia bred true with respect to mucilage canals. This characteristic is therefore considered generally unsuitable for taxonomic and biological species determination, though there may be evidence for intraspecific genotypic differentiation of Nova Scotian populations.     

Infrageneric phylogeny of the genus Gentiana (Gentianaceae) inferred from nucleotide sequences of the internal transcribed spacers (ITS) of nuclear ribosomal DNA

The internal transcribed spacers (ITSs) of nuclear ribosomal DNA have been sequenced for 20 species of Gentiana. By incorporating previously released sequence data of eight species, phylogenelic analyses using Fitch parsimony and character-state weighted parsimony were carried out. The length of ITS 1 in the taxa surveyed ranged from 223 to 238 bp and ITS2 from 216 to 234 bp. Sequence divergence between pairs of species ranged from 5.0% to 48.9% in ITS1, from 1.1% to 45.3% in ITS2, and from 3.2% to 46.1% in combined data of ITS1 and ITS2. The ITS phylogeny was generally congruent with morphological classifications except that G. asclepiadea was revealed to be closely related to section Gentiana instead of section Pneumonanthe and section Stenogyne was shown to be a paraphyletic group of the genus Gentiana that would be better excluded from the genus. A divergence among the three European endemic sections and the remaining sections of the genus other than section Stenogyne was revealed. Thus the European species of the genus together do not form a monophyletic group. A close relationship between the sections Chondrophyllae s. l. (including section Dolichocarpa), Cruciata and Pneumonanthe was suggested. The section Frigidae s. l. (including sections Monopodiae, Isomeria, Microsperma, and Phyllocalyx) contained two well-supported clades: section Frigidae s. str. and all others together. The monophyly of the typically dysploid group section Chondrophyllae s. l. was confirmed. Optimization of chromosome numbers on the ITS phylogeny suggested that 2/1 = 26 is a plesiomorphic state for the clade comprising sections Frigidae s. l., Cruciata, Pneumonanthe, and Chondrophyllae s. l., and probably 2n = 20 is a plesiomorphic state for the dysploid group, section Chondrophyllae s. l.