Structure of Phaseolus lunatus Testa at its Central Point

Tadeusz Aniszewski1*, Anna-Liisa Karttunen2, And Heikki Hyvarinen3

1Laboratory of Applied Botany, 2Laboratory of Histology, 3Laboratory of Transmission Electron Microscopy, Department of Biology, University of Joensuu,
P. O. Box 111, 80101 Joensuu, Finland

*e-mail: Tadeusz.Aniszewski@joensuu.fi

Received November 2, 2005; revision accepted January 30, 2006

In the Phaseolus lunatus seed, the strata and cells of the testa at the central point of the broad side are described. The proposed regionalization and the central point technique makes it possible to define precisely each site in the testa. Exo-, meso- and endotesta are found to be basic strata in the central point studied. The exotesta is divided into cuticle, palisade and crushed cell layers. The cuticle layer is irregular and noncellular. The palisade cells are exoepidermal sclerenchyma and the crushed cells are former exoepidermal cells. The mesotesta is composed of hourglass cells, called stapes cells here because their form resembles stapes, and crushed cells. The stapes cells are parenchyma. The endotesta is composed of support tissue, three crushed cell layers, phloem, xylem and testa bottom. The endotesta adjoins the cotyledon.

Key words: Phaseolus lunatus, endotesta, exotesta, Malpighian cells, mesotesta, legume, testa structure. 

Influence of N,N bis(3-aminopropylo)dodecyloamine of the Ultrastructure of Nuclei in Aspergillus niger Mycelium and on Cell Proliferation and Mitotic Disturbances in Allium cepa L. Root Meristem

Barbara Gabara1*, Joanna Kalwinek1, Anna Koziróg2, Zofia Żakowska2, 
and Bogumił Brycki3

1Department of Plant Cytology and Cytochemistry, 
University of Łódź, ul. Banacha12/16, 90-227 Łódź, Poland
 2Institute of Fermentation Technology and Microbiology, Technical University,
 ul. Wólczańska 171/173, 90-530 Łódź, Poland
 3Department of Chemistry, Adam Mickiewicz University, 
ul. Grunwaldzka 6, 60-780 Poznań, Poland

*e-mail: bargab@ biol.uni.lodz.pl

Received July 25, 2005; revision accepted January 25, 2006

Hyphae growing for 1 h in the presence of 0.05% and 0.1% N,N-bis(3-aminopropylo)dodecyloamine (APDA) showed malformations in the nuclear structure, including changes in shape, budding, progressive disappearance of the nucleolus and chromatin, and damage to the nuclear envelope. The Allium test revealed that at both concentrations used (0.005%, 0.01%) the tested substance lowered the frequency of dividing cells and induced mitotic abnormalities such as c-metaphases, anaphase-telophase bridges, lagging chromosomes, micronuclei, binucleate cells, budding nuclei and partial extrusion of the nucleolus from the nucleus. These data indicate that APDA has cytotoxic, mitodepressive and turbogenic effects, especially at the higher dose. The results showed that APDA may be safely used as a microbicidal agent to destroy microorganisms developing on experimental material.

Key words: Aspergillus niger, Allium cepa, nuclear ultrastructure, mycelium, root meristem, mitotic index, disturbances in mitosis, N,N-bis(3-aminopropylo)dodecyloamine. 

Distribution of Atropine and Scopolamine in Different Organs and Stages of Development in Datura stramonium L. (Solanaceae).
Structure and Ultrastructure of Biosynthesizing Cells

Alireza Iranbakhsh1*, Mohammad Ali Oshaghi2**, and Ahmad Majd 3Alireza Iranbakhsh1*, Mohammad Ali Oshaghi2**, and Ahmad Majd 3

1Department of Plant Sciences, Garmsar University of Islamic Azad, Garmsar, 
35815/144, I.R. Iran
2Department of Plant Protection, Damghan University of Islamic Azad, Damghan, I.R. Iran
3Department of Biology, Sadra University, Tehran, I.R. Iran

*e-mail: iranbakhshar@yahoo.com 
**oshaghima@yahoo.com

Received March 25, 2004; Revision accepted November 10, 2005

This paper reports a study of the alkaloid content of different parts of Datura stramonium L. (Solanaceae) in various stages of its growth and development, and the location, structure and ultrastructure of alkaloid-biosynthesizing cells (idioblasts) in different parts of naturally grown and cultured plant material. TLC, HPLC, and GC/MS methods were used for alkaloid assays. The results showed that alkaloid production starts from the end of the second week after seed germination, increases in different organs up to the tenth week of growth, and then decreases. Leaves and capsules showed the highest alkaloid content in the vegetative and generative stages, respectively. In leaves the alkaloids decreased rapidly in the generative stage. The highest alkaloid content was recorded in vegetative leaves, followed in descending order by vegetative petioles, generative and vegetative stems, generative petioles, generative roots, generative leaves, vegetative roots and mature seeds. The organs as well as calli derived from different leaf parts were examined for the presence of idioblasts by microscopic and cytochemical methods. Idioblasts were present only in semi-hyaline callus originated from leaf base; they were spherical or oval, with a thick cell wall and large central vacuole. These observations should prove helpful in attempts to produce specific alkaloids in naturally grown plants and cell cultures.

Key words: Datura stramonium, tropane alkaloids, idioblast, atropine, scopolamine.

Morphology and Anatomy of Scrophularia L. 
Taxa From NE Anatolia

Serdal Makbul1*, Kamil Coşkunçelebi2, Zafer Türkmen3, 
and Osman Beyazoğlu2

1Department of Biology, 3Faculty of Fisheries, 
Karadeniz Technical University, 53100, Rize, Turkey
2Department of Biology, Karadeniz Technical University, 
61080, Trabzon, Turkey

*e-mail: makbul@ktu.edu.tr

Received October 25, 2005; revision accepted January 20, 2006

This study uses numerical methods to describe, illustrate and assess the taxonomic significance of morphological and anatomical features of Scrophularia ilwensis C. Koch, S. capillaris Boiss. & Ball., S. nodosa L., S. libanotica Boiss. var. pontica R. Mill, S. lucida L. and S. cinerascens Boiss. collected from NE Anatolia. Features related to peculiarities of the leaf, bracteole, alar pedicel and corolla were found to be important in separating the taxa morphologically, but principal component analysis of all characters showed some anatomical characters (mean number of the epidermis, stomata, diameter of vascular bundle and scleranchymatic sheath) to be more important than morphological ones in explaining the variation between the examined taxa. 

Key words: Scrophularia, anatomy, idioblast, numerical analysis.

Diversity of Seed Coat Ultrastructure and Soluble Carbohydrate Content
 in Lupinus 

Dariusz J. Michalczyk1*, Agnieszka I. Piotrowicz-Cieślak1, Ewa Sawicka-Sienkiewicz2, Renata Galek2, Anna Bochenek1, and Katarzyna Głowacka1

1Department of Plant Physiology and Biotechnology, University of Warmia and Mazury, ul. Oczapowskiego 1A, 10-718 Olsztyn, Poland
2Department of Plant Breeding and Seed Production, 
University of Agriculture, ul. Cybulskiego 34, 50-205 Wrocław, Poland

*e-mail: darim@uwm.edu.pl

Received August 10, 2005; revision accepted January 19, 2006

Seed sculpture, seed coat thickness and soluble carbohydrate profiles were compared in three lupin species, each represented by three different genetic lines, from sections Pilosus (L. pilosus) and Atlanticus (L. atlanticus, L. cosentinii). Seed coat thickness varied most in the three lines of L. pilosus (330–1000 µm), while in the other species it varied little from 400 µm thickness. The studied seeds accumulated approximately 100 mg/g d.m. soluble carbohydrates. The obtained results together with published data suggest that the soluble carbohydrate composition of seeds is clearly section-specific in Lupinus sections Pilosus and Atlanticus. Sect. Pilosus was characterized by a high level of galactosyl cyclitols, while seeds of sect. Atlanticus contained much higher amounts of the raffinose family oligosaccharides. In all species, however, the dominating soluble carbohydrate was stachyose. The results on the biochemical parameters analyzed in this paper confirm that L. atlanticus and L. cosentinii are more closely related to each other than to L. pilosus. The seed coat roughness/smoothness criterion seems inadequate for building a classification of lupin species, as one of the analyzed lines of L. pilosus, formally ascribed to the rough-seeded lupins, produces smooth seeds.

Key words: Lupin seeds, seed sculpture, seed coat thickness, soluble carbohydrates, galactosyl cyclitols.

Immunolocalization Of Lipoxygenase in the Anther 
of Gagea lutea (L.) Ker.-Gaw.IMMUNOLOCALIZATION OF LIPOXYGENASE IN THE ANTHER OF GAGEA LUTEA (L.) KER.-GAW.

Ewa Szczuka1*, Ewa Skórzyńska-Polit2, Bożena Pawlikowska-Pawlęga3, Jolanta Sobieska4, and Antoni Gawron3 

1Department of Plant Anatomy and Cytology, 2Department of Plant Physiology, 
3Department of Comparative Anatomy and Anthropology,
 Maria Curie-Skłodowska University, Akademicka 19, 
20-033 Lublin, Poland
4Department of Biology and Horticulture, University College of Arts and Natural Science, Krakowska 26, 27-600 Sandomierz, Poland

 *e-mail: eszczuka@biotop.umcs.lublin.pl

Received August 30, 2005; revision accepted December 20, 2005

Immunogold labelling revealed the presence of lipoxygenase (LOX) in different parts and types of anther cells of Gagea lutea. LOX was found in the cytoplasm and close to ER elements in epidermal and endothecial cells, and close to the cell walls of the latter. The positive immunoreaction to LOX was less intense in the middle layers and the loculus of the anther, where single immunogold particles were concentrated at the cell walls of these layers and in the protoplast masses, in vacuoles, close to mitochondria, inside plastids, and in the liquid of the anther cavity. LOX occurred in the cytoplasm and around ER elements of pollen grains as well as in the exine layer, particularly in contact regions between the outer and inner exine layers. The correlations between LOX localization in different anther cells and the functioning of particular anther parts are discussed.

Key words: Lipoxygenase, anther, antibodies, pollen, pollen coat, Gagea lutea.

Abbreviations: LOX - lipoxygenase; LOX PAb - anti-lipoxygenase polyclonal antibody.

Embryological Analysis of Consolida regalis L.(Ranunculaceae)

Meral Ünal* and Fİlİz Vardar

Marmara University, Department of Biology, 
Göztepe, 34730 İstanbul, TurkeyGöztepe, 34730 Ýstanbul, Turkey

*e-mail: meralunal@marmara.edu.tr 

Received September 20, 2005; revision accepted January 12, 2006

In Consolida regalis L. the development of the embryo sac conforms to the Polygonum type. In the mature embryo sac, the egg apparatus and secondary nucleus are of normal appearance. The synergids are ephemeral and disappear soon after fertilization. The antipodal cells are large and persistent but remain uninucleate and show rhythmical growth. The volumes of antipodal nuclei correspond with ploidy levels from n to 256n. Cytochemical tests indicate the presence of high amounts of proteins, lipids, polysaccharides, RNA and DNA in the antipodals. They degenerate in the cellular endosperm stage. The endosperm is nuclear; centripetal wall formation starts at the micropylar end and progresses towards the chalaza. Embryogeny is of the Solanad type. The mature embryo lacks organs at shedding of seeds.

Key words:  Consolida regalis, Ranunculaceae, embryo sac, antipodals, endoreduplication, embryo, endosperm.

Geographic Patterns of Diversity of Cultivated Lentil Germplasm Collected 
from Pakistan, as Assessed by Seed Protein Assays 

Tayyaba Sultana1, Abdul Ghafoor2*, and Muhammad Ashraf1

1Department of Biological Sciences, Quaid-i-Azam University, Islamabad, Pakistan
2Plant Genetic Resources Programme, National Agricultural Research Centre, Islamabad, Pakistan

*e-mail: ghafoor59pk@yahoo.com

Received July 15, 2005; revision accepted February 15, 2006

Seed protein profiles were studied in 144 lentil accessions intensively collected from all over Pakistan. Heterogeneous populations were isolated on the basis of SDS-PAGE, and 13 polymorphic protein peptides were found, representing almost all the variation reported so far in lentil. The low diversity of accessions from the Northern Area and North Western Frontier Province, the most geographically diverse areas, suggested the need for more exploration so that the maximum genetic diversity of the areas can be truly represented. Clusters based on agro-ecological zones did not prove adequate for evaluation of lentil resources, whereas 63 of 108 accessions (58.3%) were grouped together by altitude and provincial distribution. The study confirmed the wealth of phenotypic divergence in the local lentil. A small sample of accessions from a particular region might not reflect the actual diversity within that region. Samples representing total diversity in particular countries or regions should be evaluated, so that a representative rather than a random set of accessions can be included in investigations of diversity on regional or continental scales. As Pakistan is in the vicinity of the centre of diversity of lentil, high variation of various parameters is expected, and that can be found only if a complete set of germplasm is studied.

Key words: Lens culinaris, cluster analysis, genetic diversity, landraces, SDS-Polyacrylamide Gel Electrophoresis. 

Pollen Morphology of Some Turkish Crocus L. (Iridaceae) Species

Serap Işık1* and Emel Oybak Dönmez2**

1Hacettepe University, Department of Secondary Science and Mathematics Education, 2Department of Biology, 06800, Beytepe, Ankara-Turkey

*e-mail: serapi@hacettepe.edu.tr
**e-mail: polen@hacettepe.edu.tr

Received October 21, 2005; revision accepted March 15, 2006

Pollen grains of 29 Turkish taxa of genus Crocus L. belonging to Iridaceae were examined by light and electron microscopy. The pollen of Crocus is spheroidal in shape, with a thin exine characterized by echinate (spinulate)-microperforate sculpture and a relatively thick intine. Three types of aperture are recognized: a spiral furrow, more or less extensive furrows, and short furrows. Spiral furrows appear to be the most frequent aperture type among the Turkish Crocus species. Apertures vary within some species. The results are compared with those on the pollen morphology of Crocus species previously investigated by several European workers.

Key words: Crocus, Iridaceae, pollen morphology, Turkey.

Cytomorphological studies on American and European 
Phleum commutatum Gaud. (Poaceae)

Adam Kula1, Barbara Dudziak4, Elwira Sliwinska2, Aleksandra Grabowska-Joachimiak1, Alan Stewart3, Hieronim Golczyk4 and Andrzej J. Joachimiak4*

1Cytogenetics Group in the Department of Plant Breeding and Seed Science, The Agricultural University of Cracow, Łobzowska 24, 31-140 Kraków, Poland
2 Laboratory of Molecular Biology and Cytometry, Department of Genetics and Plant Breeding, University of Technology and Agriculture, 85-796 Bydgoszcz, Poland
3 Ceres Research Centre, PO Box 3100, Christchurch 8015, New Zealand
4Department of Plant Cytology and Embryology, Institute of Botany, Jagiellonian University, 
ul. Grodzka 52, 31-044 Kraków, Poland

*corresponding author, email: a.joachimiak@iphils.uj.edu.pl

Received March 25, 2006; revision accepted May 10, 2006

The morphology, karyotype structure and nuclear DNA amount of European and American representatives of alpine cat’s tail Phleum commutatum Gaud. were analysed. The evolutionary relationships within this group of taxa are discussed.
It was shown that the American and European tetraploids are both morphologically and karyologically similar to each other and to the diploid race of this species from the Polish Carpathians.
Diploid relatives of tetraploid P. commutatum i.e., P. commutatum (2x) and P. rhaeticum (Humphries) Rauschert, showed intra- and interspecific differences in nuclear 2C DNA values, whereas tetraploids (of both European and American origin) showed highly uniform nuclear DNA amounts. The 2C DNA uniformity and the greater mean size of the basal chromosome set in tetraploid P. commutatum as compared with its diploid relatives suggest that the change in genome size occurred during early evolution of the tetraploid race of P. commutatum, before its migration to America.

Key words: Phleum commutatum, P. alpinum, P. rhaeticum, morphology, chromosomes, karyotype, C-banding, heterochromatin, flow cytometry, nuclear genome size.

Chromium Accumulation and its Effects on Other Mineral Elements 
in Amaranthus viridis L.

Jinhua Zou1, Min Wang1, Wusheng Jiang2, And Donghua Liu1*

1Department of Biology, College of Chemistry and Life Sciences,
Tianjin Normal University, Tianjin 300074, ChinaTianjin Normal University, Tianjin 300074, China
2Library, Tianjin Normal University, Tianjin 300074, P.R. China2Library, Tianjin Normal University, Tianjin 300074, P.R. China

*e-mail: hsxyldh@mail.tjnu.edu.cn

Received July 18, 2005; revision accepted February 15, 2006

Chromium accumulation and its effects on other mineral elements in Amaranthus viridis L. were investigated using inductively coupled plasma atomic emission spectrometry (ICP-AES) and resin adsorption. The aim was to understand why A. viridis can grow well in soils heavily contaminated by Cr, what the forms of Cr in soils and residues are, and what effects Cr has on the distribution of Mn, Fe, Cu and Zn in the plant. The results indicated that A. viridis is not a hyperaccumulator, although it can grow well in soil containing a high concentration of Cr. The Cr concentration in plant tissues from a contaminated site was about 11 times higher than in those from an uncontaminated site. At both the contaminated and uncontaminated sites, Cr was accumulated primarily in its shoots, and in roots in much lower concentrations. The levels of Cr in A. viridis tissues were as follows: leaf > root > stem. Cr occurred predominantly as Cr(III) there was very little Cr(VI) in the polluted soil where A. viridis grew close to a waste heap discarded by a chromium(VI) production factory. The possible mechanisms for Cr accumulation of A. viridi and the effects of Cr on uptake and accumulation of the other mineral elements in A. viridi are briefly discussed.

Key words: Amaranthus viridis L., chromium, copper, iron, manganese, zinc, hyperaccumulation.

Genetic Analysis of Tomato (Lycopersicon esculentum Mill.) Twin Forms

Lubosława Nowaczyk and Paweł Nowaczyk 

Department of Genetics and Plant Breeding, University of Technology and Agriculture, Department of Genetics and Plant Breeding, University of Technology and Agriculture, 
ul. Bernardynska 6, 85-029 Bydgoszcz, Poland ul. Bernardynska 6, 85-029 Bydgoszcz, Poland 

e-mail: warz@atr.bydgoszcz.pl

Received October 20, 2005; revision accepted January 31, 2006

The aim of the study was to determine the origin of additional embryos on the basis of marker characters observed in the phenotype of twin pairs of F2 and F3 generations of two hybrid cultivars of tomato (Alonso and Carmello). The marker characters were the presence/absence of the so-called green back, and the type of growth which can be indeterminate or determinate. Among 112 F2 twins, differences in marker characters were observed in 24 pairs. In the material derived from Alonso, 9 of 18 observed F3 progenies did not show segregation in marker characters. Progenies of Carmello had higher variability; only 12 of 28 progeny groups showed no recombinants. Analysis of the F3 generation enabled further limitation of the sources of additional embryos. Segregation of any marker character excluded the possibility that a particular F2 twin was formed from a haploid cell of the embryo sac after spontaneous doubling of chromosome number.

Key words: Adventitious embryogenesis, apomictic polyembryony, marker gene, splitting polyembryony.

Influence of exogenous carbohydrates on superoxide dismutase activity in Trifolium repens L. explants cultured in vitro

Ireneusz Ślesak 1*, Wojciech Hałdaś 2, and Halina Ślesak 2

1Institute of Plant Physiology, Polish Academy of Sciences,
ul. Niezapominajek 21, 30-239 Cracow, Poland
2 Department of Plant Cytology and Embryology, 
Jagiellonian University, ul. Grodzka 52, 31-044 Cracow, Poland

*e-mail: i.slesak@ifr-pan.krakow.pl

Received November 24, 2005; revision accepted April 25, 2006

The effect of four sugars (sucrose, glucose, fructose, maltose) used as carbon source in liquid medium on superoxide dismutase (SOD) activity was studied. Little is known about how different soluble sugars used in plant tissue culture can affect SOD activity, an antioxidant enzyme involved in regulating the level of reactive oxygen species (ROS) in plant cells. In extracts of 15-day old cotyledons and hypocotyls of Trifolium repens L., four SOD forms were detected by native PAGE and activity staining: MnSOD, FeSOD, Cu/ZnSOD I and Cu/ZnSOD II. Activity of FeSOD and Cu/ZnSOD II was significantly lower in cotyledons cultured on media supplied with different carbohydrates than in control cotyledons cultured on medium without sugars, or was absent altogether. FeSOD was absent in hypocotyls obtained from seedlings grown on sterile filter paper (SFP); those cultured on sucrose-supplemented medium showed weak FeSOD activity. FeSOD appeared to be the most labile SOD form in T. repens cotyledons. MnSOD activity was highest in cotyledons cultured on sucrose-, fructose- and maltose-based media. Cu/ZnSOD I activity was highest in cotyledons cultured on medium with glucose and fructose. Sucrose strongly inhibited both FeSOD and Cu/ZnSOD II activity. The results indicated that various carbohydrates affect the activity of SOD forms in T. repens cotyledons. The decrease in activity of FeSOD and Cu/ZnSOD II, putatively located in chloroplasts, might suggest an inhibitory effect of some carbohydrates on photosynthesis in cotyledons of T. repens. On the other hand, supply of sucrose, fructose and maltose increased the activity of the putative mitochondrial form of MnSOD. This result might indicate sugar-induced production of ROS during respiration in mitochondria.

Key words: Trifolium repens, plant tissue culture, SOD pattern.

Factors Affecting Seed Germination and Seedling Growth 
of Terrestrial Orchids Cultured 

HANA VEJSADOVÁ

Department of Landscape Biodiversity, 
Silva Tarouca Institute for Landscape and Ornamental Gardening, 
252 43 Průhonice, Czech Republic

Received: August 10, 2005; revision accepted February 12, 2006

The effect of two sterilization substances (calcium hypochlorite and sodium hypochlorite) on the germination rate and the effect of nitrogen and growth regulators on seedling growth and development was studied in three critically endangered species of terrestrial orchids (Dactylorhiza incarnata subsp. serotina, Dactylorhiza maculata subsp. maculata, Liparis loeselii). Surface sterilization of mature seeds using 7.2% calcium hypochlorite (until decolorization from brown to ivory) stimulated the germination rate. Addition of peptone at 1 g l-1 concentration or auxin 3-indoleacetic acid (IAA; 1.43 µM) with cytokinin zeatin (0.72 µM) and 1-naphthylacetic acid (NAA; 1.34 µM) to the culture medium significantly increased the growth parameters of seedlings after 12 months.

Key Words: Terrestrial orchids, in vitro generative propagation, seed sterilization, nitrogen form, growth regulators.