Nuclear DNA and heterochromatin contents in theScilla hohenackeri group,S. persica,andPuschkinia scilloides (Liliaceae)

DNA contents (presented as 1C-values) have been determined cytophotometrically in 7 species of theScilla hohenackeri group (10.18 to 22.71 pg), and in the systematically more isolated taxaS. persica (21.02 pg) andPuschkinia scilloides (6.80 pg). The heterochromatin amount is not correlated with the nuclear DNA content. Euchromatin, therefore, is not a particularly conservative part of the genome. However, high C-values and large but few terminal heterochromatin bands, and lower C-values and numerous but smaller heterochromatin bands are found to be linked in theS. hohenackeri group. Obviously, numerous chromosomal changes have accompanied speciation in this group. DNA contents, and C-banded karyotypes are consistent with systematic affinities based on morphological similarities.Evolution ofScilla and Related Genera, III.     

Studies on systematic embryology inScilla (Hyacinthaceae)

Scilla persica and 5 species of the so-calledS. hohenackeri group, namely,S. furseorum, S. puschkinioides, S. vvedenskyi, S. hohenackeri, andS. greilhuberi, have been investigated embryologically with special reference to embryo sac and endosperm development.Polygonum-type embryo sac development was stated inS. puschkinioides andS. greilhuberi. 8-nucleate, normally structured embryo sacs, which could not be specified further due to sparse availability of the material, were stated inS. furseorum, S. vvedenskyi, andS. hohenackeri. InS. persica the embryo sac develops according to the bisporicAllium-type. In most species endosperm development was stated to be nuclear, exceptS. hohenackeri, where the type could not be specified. Other traits of possible taxonomic significance are the number of layers in the outer integument, which is mostly 4, or 5–6 inS. furseorum, and the occurrence of polyploid versus haploid and early degenerating antipodal nuclei, the latter occurring only inS. persica andS. furseorum. These embryological characters may be useful for assessing taxonomic relationship of the present species with other allied groups withinScilla, in particular, theS. siberica alliance,S. messeniaca, and theS. bifolia alliance. TheAllium-type embryo sac, which occurs inS. persica, is also characteristic for theS. siberica alliance, and may be a common derived character. Lack of antipodal polyploidization, as characteristic forS. persica andS. furseorum, occurs also in theS. siberica alliance, and is perhaps another common derived trait indicating phylogenetic relationship. Nuclear endosperm development is more frequent in spring-flowering squills than helobial development, which has previously been stated inS. messeniaca, some species of theS. siberica alliance, and inS. litardierei. While helobial endosperm may be primitive forHyacinthaceae in general, it may, by reversal, also occur as a derived character, at least in some species of theS. siberica alliance.     

A new polyploidScilla (Liliaceae) from the Cretan area (Greece)

Scilla talosii Tzanoudakis etKypriotakis from the islet of Dia is described as a new species. It is an autumn-flowering taxon related toScilla autumnalis L. andS. obtusifolia Poiret but distinct morphologically and cytologically from both these taxa.S. talosii is polyploid with 2n=ca. 150 chromosomes.     

A Giemsa C-banding and DNA content study inScilla cilicica andS. morrisii,two little known sibling species of theS. siberica alliance (Hyacinthaceae)

S. cilicica Siehe andS. morrisii Meikle are two little known sibling species of theScilla siberica alliance with previously difficult taxonomy, especially with regard to specific delimitation and geographic distribution. Quantitative C-band karyotyping and DNA content determinations were performed in four provenances, including the type localities. Both species (2n = 12) differ strikingly in their banding patterns despite of similar DNA content. Karyotype diversification involved changes in eu- and heterochromatin quantities and resulted in some changes of chromosome form. Despite this, actual phylogenetic relatedness is indicated by a chromosomal marker (NOR-position) of rare occurrence in the alliance. Karyological results and circumstantial evidence from the literature suggest thatS. morrisii is an endemic, exclusive to Cyprus as a member of theS. siberica alliance.S. cilicica seems to be restricted to continental S. Anatolia opposite to Cyprus. Its occurrence on Cyprus is questionable.The authors dedicate this paper respectfully to emer. O. Univ.-Prof. DrLothar Geitler on the occasion of his 90th birthday.     

TheCBP2 gene fromSaccharomyces douglasii is a functional homologue of theSaccharomyces cerevisiae gene and is essential for respiratory growth in the presence of a wild-type (intron-containing) mitochondrial genome

InSaccharomyces cerevisiae the only known role of theCBP2 gene is the excision of the fifth intron of the mitochondrialcyt b gene (bI5). We have cloned theCBP2 gene fromSaccharomyces douglasii (a close relative ofS. cerevisiae). A comparison of theS. douglasii andS. cerevisiae sequences shows that there are 14% nucleotide substitutions in the coding region, with transitions being three times more frequent than transversions. At the protein level sequence identity is 87%. We have demonstrated that theS. douglasii CBP2 gene is essential for respiratory growth in the presence of a wild-typeS. douglasii mitochondrial genome, but not in the presence of an intronlessS. cerevisiae mitochondrial genome. Also theS. douglasii andS. cerevisiae CBP2 genes are completely interchangeable, even though the intron bI5 is absent from theS. douglasii mitochondrial genome.     

Anthelmintic,anticoccidial and antioxidant activity of Salvadora persica root extracts

Human infection with parasites is still one of the big problems worldwide. Medicinal plants succeeded to overcome a variety of protozoan and helminthic parasites. In this study, Salvadora persica root extracts (SE) were used to treat helminthosis and coccideosis. Three doses were used (200, 100 and 50 mg/ml) to study the anthelmintic activity of S. persica. Allolobophora caliginosa was used as a model worm. Also, Albendazole was used as a reference drug. In order to study the anticoccideal activity of SE, a group of mice were infected with Eimeria papillata sporulated oocysts. Experimental mice were treated with SE (300 mg/Kg) for 5 days. The extract was able to decrease the number of meronts and gamonts of the parasite in jejunum. Also, it regulates the level of glutathione and malondialdehyde and the activity of catalase as well. We conclude that S. persica possesses a powerful Anthelmintic, anticoccidial and antioxidant activity.     

Beitrag zur systematik vonScilla L. subgen.Scilla (inklusiveChionodoxa Boiss.)

Zusammenfassung Die vonBoissier aufgestellte GattungChionodoxa wird auf Grund samenanatomischer und karyologischer Befunde eingezogen. Ihre nahe Verwandtschaft mitScilla bifolia agg. konnte aufgezeigt werden. Die wesentlichen Unterschiede derScilla bifolia- Gruppe (inklusiveChionodoxa-Arten) zu anderen Gruppen myrmekochorerScilla-Arten sowie zuPuschkinia undHyacinthus wurden dargelegt.Folgende neue Kombinationen werden aufgestellt:Scilla luciliae (Boiss.)Speta comb. nova,Scilla cretica (Boiss. &Heldr.)Speta comb. nova,Scilla nana (Schult. fil.)Speta comb. nova,Scilla sardensis (Barr &Sugden)Speta comb. nova,Scilla siehei (Stapf)Speta comb. nova,Scilla×allenii (Nicholson)Speta comb. nova.Weiters wurden die Chromosomenzahlen für einigeScilla- Arten überprüft und dabei fürScilla bifolia neben der bekannten diploiden auch die tetraploide Zahl 2n=36 aufgefunden.

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Summary The genusChionodoxa set up byBoissier is withdrawn due to seedanatomical and caryological statements and its species are incorporated intoScilla L. The close affinity between formerChionodoxa-species andScilla bifolia agg. could be demonstrated. The essential differences between theScilla bifolia group (including the formerChionodoxa species) and other groups of myrmecochorousScilla species,Puschkinia andHyacinthus were shown. The following new combinations are set up:Scella luciliae (Boiss.)Speta comb. nova,Scilla cretica (Boiss. &Heldr.)Speta comb. nova,Scilla nana (Schult. fil.)Speta comb. nova,Scilla sardensis (Barr &Sugden)Speta comb. nova,Scilla siehei (Stapf)Speta comb. nova,Scilla×allenii (Nicholson)Speta comb. nova.Furthermore the chromosome numbers of someScilla species were checked, and inScilla bifolia L. besides the known diploid number the tetraploid number 2n=36 was discovered.

     

Sequence and characterisation of a ribosomal RNA operon fromAgrobacterium vitis

One of the four ribosomal RNA operons (rrnA) from theAgrobacterium vitis vitopine strain S4 was sequenced.rrnA is most closely related to therrn operons ofBradyrhizobium japonicum andRhodobacter sphaeroides and carries an fMet-tRNA gene downstream of its 5S gene, as in the case ofR. sphaeroides. The 16S rRNA sequence of S4 differs from theA. vitis K309 type strain sequence by only one nucleotide, in spite of the fact that S4 and K309 have very different Ti plasmids. The predicted secondary structure of the S4 23S rRNA shows several features that are specific for the alpha proteobacteria, and an unusual branched structure in the universal B8 stem. The 3′ ends of the three otherrrn copies of S4 were also cloned and sequenced. Sequence comparison delimits the 3′ ends of the four repeats and defines two groups:rrnA/rrnB andrrnC/rrnD.     

AgTHR4, a new selection marker for transformation of the filamentous fungusAshbya gossypii,maps in a four-gene cluster that is conserved betweenA. gossypii andSaccharomyces cerevisiae

Single-read sequence analysis of the termini of eight randomly picked clones ofAshbya gossypii genomic DNA revealed seven sequences with homology toSaccharomyces cerevisiae genes (15% to 69% on the amino acid level). One of these sequences appeared to code for the carboxy-terminus of threonine synthase, the product of theS. cerevisiae THR4 gene (52.4% identity over 82 amino acids). We cloned and sequenced the complete putativeAgTHR4 gene ofA. gossypii. It comprises 512 codons, two less than theS. cerevisiae THR4 gene. Overall identity at the amino acid sequence level is 67.4%. A continuous stretch of 32 amino acids displaying complete identity between these two fungal threonine synthases presumably contains the pyridoxal phosphate attachment site. Disruption of theA. gossypii gene led to threonine auxotrophy, which could be complemented by transformation with replicating plasmids carrying theAgTHR4 gene and variousS. cerevisiae ARS elements. Using these plasmids only very weak complementation of aS. cerevisiae thr4 mutation was observed. Investigation of sequences adjacent to theAgTHR4 gene identified three additional ORFs. Surprisingly, the order and orientation of these four ORFs is conserved inA. gossypii andS. cerevisiae.     

HLA-D typing in 111 multiple sclerosis patients: Distribution of four HLA-D alleles

Multiple sclerosis patients-111 of them-were typed for theHLA-D allelesw1, w2, andw3 and a new determinant, EI. Statistically significant increase was obtained for thew2 andw3 frequencies as compared to healthy controls. The distribution of HLA-D phenotypes among MS patients revealed a good fit according to the Hardy-Weinberg law. By calculating linkage disequilibrium parameters, theHLA-B7,Dw2 allele combination was found to be more closely associated than in normal controls, whereas forHLA-Bw35, Dw1, no linkage disequilibrium could be detected in the patients' group. From these data we conclude that in multiple sclerosis, the disturbance affects the frequency ofDw3 and the gametic association between alleles of theHLA-B andD loci, as well as the already known increase ofHLA-B7 andDw2.     

Giemsa karyotypes and their evolutionary significance inScilla bifolia,S. drunensis,andS. vindobonensis (Liliaceae)

Quantitatively evaluated C-banded and conventional karyograms are presented for the related speciesScilla bifolia subsp.danubialis Speta,S. drunensis (Speta)Speta, andS. vindobonensis Speta. On the basis of banding patterns and karyotype structureS. bifolia subsp.danubialis (2n = 18, 2×) andS. drunensis (2n = 36, 4×) are quite similar, whileS. vindobonensis (2n = 18, 2×) is entirely different. There is a moderate degree of karyotypic variation withinS. bifolia subsp.danubialis andS. drunensis. However, inS. vindobonensis karyotypes and banding patterns are almost completely stable over a geographical range of about 500 km. The present results confirm the recent taxonomic separation ofS. vindobonensis fromS. bifolia, and suggest a considerable phylogenetic distance between these two diploid species. The results are discussed with reference to the morphological characters of the species and their geographical distribution.Evolution ofScilla and Related Genera, II.Dedicated to Univ.-Prof. Dr.Elisabeth Tschermak-Woess on the occasion of her 60th birthday.     

Effects of Iso-osmotic Concentrations of NaCl and Mannitol on some Metabolic Activity in Calluses of Two Salicornia species

The comparative effects of salinity and drought on activity of antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POX), catalase (CAT), and polyphenol oxidase (PPO) and contents of protein, proline, hydrogen peroxide, malondialdehyde (MDA), and carotenoids were studied in calluses of Salicornia persica and Salicornia europaea. Calluses were cultured on Murashige and Skoog medium (MS medium) containing iso-osmotic concentrations of NaCl (100, 300, and 600 mM) and mannitol (200, 500, and 1000 mM). Proline content increased in calluses of both species under NaCl- and mannitol-induced stresses. Under both stresses, S. persica accumulated higher amounts of protein as compared to S. europaea. Under both stresses, protein content in S. persica decreased in comparison to control. Antioxidant enzyme activities showed different trends in the two Salicornia species under salt and drought stresses but were higher in S. europaea than in S. persica. Changes of hydrogen peroxide and MDA contents of calluses subjected to salt stress followed a pattern different from that observed under drought stress. MDA content showed similar changes to hydrogen peroxide content. Carotenoid content decreased in both species under both stresses. Overall, at the cellular level, S. europaea calluses were more tolerant to both NaCl- and mannitol-induced stresses than calluses of S. persica.     

In Vitro Scolicidal Effects of Salvadora persica Root Extract against Protoscolices of Echinococcus granulosus

It has been known that Arak, Salvadora persica, has a number of medicinal properties. We tried to investigate in vitro scolicidal effect of root extracts of this plant against protoscolices from hydatid cysts of Echinococcus granulosus. Protoscolices were aseptically collected from sheep livers containing hydatid cysts. S. persica root extract was used in 10, 30, and 50 mg/ml concentration for 10, 20, and 30 min. The viability of protoscolices was ascertained by 0.1% eosin staining. Scolicidal activity of S. persica extract at a concentration of 10 mg/ml was 36.3%, 50.3%, and 70.8% after 10, 20, and 30 min of exposure, respectively. The scolicidal effect of this extract at a concentration of 30 mg/ml was 52.9%, 86.7%, and 100% after 10, 20, and 30 min of exposure, respectively. S. persica extract at a concentration of 50 mg/ml, meanwhile, killed 81.4%, 100%, and 100% of protoscolices after 10, 20, and 30 min, respectively. Also, the cytotoxic potential of S. persica was assessed on human liver cells (HepG2) using trypan blue exclusion test. No cytotoxic effect was observed on HepG2 cell line. The present study confirmed for the first time that the ethanolic extract of S. persica has high scolicidal power in vitro. However, in vivo effect of this material remains to be studied for treatment of echinococcosis in humans and herbivorous animals.