ABSTRACTS Volume 43, 2001

Changes in Phenolic Acids of Cherry Laurel (Laurocerasus officinalis Oxygemmis) Fruit During Maturation

  F. Ahmet Ayaz

Department of Biology, Karadeniz Technical University, 61080 Trabzon, Turkey

 Received: June 27, 2000; accepted February 14, 2001

Changes in phenolic acid content during the maturation of cherry laurel (Laurocerasus officinalis Oxygemmis) fruits were studied. Samples were obtained starting 40 days after flowering between May and July. In the fruits, p-coumaric, caffeic and ferulic acids (as cinnamics) and benzoic, 4-hydroxybenzoic, vanillic and 3,4-dihydroxybenzoic acids (as benzoics) were identified. The content of benzoic, 4-hydroxybenzoic, vanillic and caffeic acids started to increase from 6 to 12 WAF (weeks after flowering). The content of p-coumaric and ferulic acids, highest at 6 WAF, declined gradually until 11 WAF and increased again at 12 WAF. All phenolic acids except vanillic and ferulic acids began to decrease in the fruits harvested at 16 WAF, that is, the normal harvesting season. Although an increase in vanillic and ferulic acid content was observed in post-ripening between 13 and 16 WAF, the values were not statistically significant (P = 0.05) between stages. The highest content of phenolic acid in the cultivar was found for benzoic, caffeic and vanillic acids: 2.53, 1.05 and 0.86 mg/100g dry weight, respectively.

Key words: Cherry laurel, cultivar, phenolic acid, fruit, maturation.


Chromosome Number of Some Alchemilla L. (Rosaceae) Species 

Sema Hayirlioglu-Ayaz* and Hüseyin Inceer

 Department of Biology, Karadeniz Technical University, 61080Trabzon, Turkey 

Received February 13, 2001; Revision accepted June 11, 2001

 This paper presents the results of karyological analysis of five Alchemilla species collected from northeast Anatolia, Turkey, belonging to sect. Alchemilla Rothm. subsect. Chirophyllum Rothm. ser. Sericeae Bus., subsect. Heliodrosium Rothm. ser. Vulgares Bus. and subsect. Calycanthum Rothm. ser. Elatae Rothm. The following chromosome numbers were determined: A. amoena (Czeczott) Rothm. 2n = 90-102, A. chlorosericea Bus. 2n = 92-103, A. glabricaulis Lindb. 2n = 90-109, A. hessii Rothm. 2n = 88-96, A. porrectidens Juz. 2n = 86-96. The chromosome numbers of three of these five species are presented for the first time.

Key words: Alchemilla, chromosome numbers, Turkey.

Glandular Apparatus Structure and Essential Oil Constituents of Satureja cuneifolia Ten.

 Nada Bezić *1, Valerija Dunkić 1, Ani Radonić2

 1University of Split, Department of Biology, Teslina 12, 21000 Split, Croatia
University of Split, Department of Organic Chemistry, Teslina 10/V, 21000 Split, Croatia

Received April 2, 2001; accepted July 6, 2001

 This study examines the anatomical structure of the glandular apparatus in Satureja cuneifolia Ten., as well as the chemical composition of the essential oil derived from it. Leaves were collected from a population in Dalmatia (Croatia). Anatomically, the glandular scales feature a unicellular base, a unicellular stalk and a 12-celled head. Each glandular apparatus was observed to be surrounded by epidermal cells. Qualitative and quantitative GC-MS analysis of the essential oil showed that linalool, carvacrol and p-cymen were its main constituents.

  Key words: Satureja cuneifolia, glandular apparatus, essential oil, linalool.

Augmenting In Vitro Shoot Multiplication with Growth Regulators and Light Conditions in Musa acuminata cv. Dwarf Cavendish

 G.R.Rout1, S.Samantaray2 and P. Das1

1Plant Biotechnology Division, Plant Tissue Culture Laboratory,
Plant Physiology and Biochemistry Laboratory, 
Regional Plant Resource Centre, Bhubaneswar 751 015, India

 Received March 1, 2000; accepted January 4, 2001

An efficient protocol was developed for in vitro mass propagation of the commercial cultivar Musa acuminata cv. Dwarf Cavendish through shoot apices culture. Shoot apices with 2-3 pairs of leaf primordia were induced from sliced rhizome explants on Murashige and Skoog (1962) medium supplemented with 6.0 mg/l BA, 150 mg/l Ads and 3% (w/v) sucrose. Multiple shoots were induced from meristematic domes on the same medium. Addition of 1.0-1.5 mg/l IAA to the culture medium and incubation in continuous light (24 h) increased the yield of multiple shoots. The high multiplication coefficient was maintained stable up to the 5th subculture; thereafter it declined. Rooting was readily achieved upon transferring the shoots onto half-strength MS medium supplemented with 500 mg/l activated charcoal and 2% (w/v) sucrose. Micropropagated plantlets were hardened in a polyethylene house and successfully established in soil. There was no morphological variation among the micropropagated plants.

Key words: Musa, growth regulators, micro-propagation, in vitro, light conditions.


Restriction Profiles of Plasmid DNA from Wild Strains of Desulfovibrio desulfuricans

 Zofia Dzierżewicz, Beata Cwalina, Joanna Szczerba, Ilona Bednarek, Urszula Mazurek, and Tadeusz Wilczok.

 Department of Molecular Biology, Biochemistry and Biopharmacy 
Medical University of Silesia, ul. Narcyzów 1, 41-200 Sosnowiec

 Received June 6, 2001; accepted August 13, 2001

One of many strategies of bacteria survival in a contaminated environment is plasmid-encoded resistance to toxic substances. D. desulfuricans wild strains isolated from soil and mud samples taken from contaminated sites were studied to identify plasmids in these bacteria and to examine their restriction profiles. The presence of plasmid DNA in all tested strains was demonstrated. Using BstEII-digested DNA of phage l as the molecular standard, plasmid size was established at ~2.3 kbp. The restriction fragments obtained after cutting plasmid DNA with endonuclease HaeIII or AluI were better separated on gradient polyacrylamide gel than on homogeneous gel. Very high similarity between the fragment profiles was demonstrated, despite the different origins of the strains tested.

 Key words: Desulfovibrio desulfuricans, sulphate-reducing bacteria, plasmid, electrophoresis, restriction fragments.


Cell Cycle Synchronization in the Root Meristem of Allium cepa L.

 Monika Wołoszyn1, Hieronim Golczyk1 and Andrzej Joachimiak* 1,2

 1Department of Plant Cytology and Embryology, JagiellOnian University, ul. Grodzka 52, 31-044 Cracow, Poland
Cytogenetics Group in the Department of Plant Breeding and Seed Science
The Agricultural University of Cracow, ul. Łobzowska 24, 31-140 Cracow, Poland

 Received May 15, 2001; accepted June 27, 2001

A high level of cell cycle synchronization in Allium cepa was achieved using a 24 h hydroxyurea block (1.25 mM) followed by 20 h of recovery (without hydroxyurea) and a subsequent 4 h treatment with saturated a-bromonaphthalene solution. This procedure was very effective in producing a synchronously dividing cell population with a 76% mean mitotic index and a 30% frequency of metaphase stages. After release from hydroxyurea arrest, large fractions of cells synchronously entered successive phases: G1, S, and G2. The calculated duration of the cell cycle was 13 h, in general accordance with previous reports. A detailed protocol for obtaining a high mitotic index and a large portion of cells in a particular cycle phase (G1, S, G2) was prepared.

Key words: Allium cepa, root meristem, cell cycle, synchronization, mitotic index, hydroxyurea, a-bromonaphthalene, DNA measurements.

RAPD studies in Bromus (Poaceae) from the Old and New Worlds - preliminary results

Andrzej Joachimiak*1,2, Agnieszka Sutkowska2, Józef Mitka3

1Department of Plant Cytology and Embryology, Institute of Botany Jagiellonian University, ul. Grodzka 52, 31-044 Cracow, Poland
Cytogenetics Group in the Department of Plant Breeding and Seed Science
The Agricultural University of Cracow, Łobzowska 24, 31-140 Cracow, Poland
Botanical Garden, Jagiellonian University, ul. Kopernika 27, 31-501 Cracow, Poland

    Received April 30, 2001; accepted July 9, 2001

RAPD analysis was applied to nine Bromus species (B. fasciculatus, B. arvensis, B. scoparius, B. hordeaceus, B. squarrosus, B. carinatus, B. willdenowii, B. erectus, B. inermis) belonging to four subgenera: subg. Stenobromus, subg. Bromus, subg. Ceratochloa, and subg. Festucaria. With 10 out 13 primers a considerable degree of polymorphism was detected at the interspecific level, and some bands obtained with these primers appeared to be species-specific. The RAPD band distribution and genetic relationships between different Bromus species and subgenera were analyzed. No bands common to all analyzed species were discovered, but some of the amplified DNA fragments were common to all taxa belonging to the same subgenera or even different ones. The divisions suggested by taxonomists generally were confirmed by numerical analysis of the RAPD data presented here.

Key words: Bromus L., nuclear DNA, RAPD, molecular markers.


Intrapopulational and Interpopulational Relations of Betula pendula Roth (Betulaceae) in Croatia, Based on Leaf Morphometry

Sanja Kovačić1* and Diana Šimić2

 1Botanical Department and Botanical Garden, University of ZagrebMarulićev trg 9a, 10000 Zagreb, Croatia
2Institute for Occupational Health, Ksaverska cesta 2, 10000 Zagreb

  Received May 7, 2001; accepted July 5, 2001

Seven morphometric variables of silver birch (Betula pendula Roth) leaves from 5 bioclimatic regions of Croatia were analyzed to establish the pattern of intrapopulational and interpopulational relations. As introgression in Croatia could be almost entirely excluded, this research may represent a model of a consistent B. pendula species. Leaf samples were taken from fertile and sterile shoots separately, and the analyses yielded statistically significant differences between them. There was significant variability of all variables within and between populations. The most prominent variables in evaluation of interpopulational variability of the leaves from both fertile and sterile shoots of B. pendula were the distance from the leaf base to the widest point of the lamina (at P = 0.01) and the angle of the leaf base (at P = 0.001). These variables could be used in future research on genetic diversity among populations of B. pendula.

Key words: Betula pendula, morphometry, leaf morphology, multivariate analysis, fertile and sterile shoots, Croatia.

C-Banding and Nuclear DNA Amount in Six Bromus Species

Andrzej Joachimiak*1,2, Adam Kula2, Elwira Śliwińska3, Anna Sobieszczańska2

1 Department of Plant Cytology and Embryology, Institute of Botany, Jagiellonian University ul. Grodzka 52, 31-044 Cracow, Poland
Cytogenetics Group in the Department of Plant Breeding and Seed Science The Agricultural University of Cracow, ul. Łobzowska 24, 31-140 Cracow, Poland
Department of Genetics and Plant Breeding, University of Technology and Agriculture
, ul. Kaliskiego 7, 85-796 Bydgoszcz, Poland

    Received May 17, 2001; accepted July 17, 2001

Conventional and C-banding chromosome studies and nuclear DNA estimations were performed on six Bromus species (B. arvensis, 2n = 2x = 14; B. hordeaceus, 2n = 4x = 28; B. carinatus, 2n = 8x = 56; B. willdenowii, 2n = 6x = 42; B. erectus, 2n = 8x = 56; and B. inermis, 2n = 8x = 56) belonging to three subgenera: subg. Bromus, subg. Ceratochloa and subg. Festucaria. Three species (B. carinatus, B. erectus and B. inermis) show different chromosome numbers within root-tip meristem. Root-tip cells of these high polyploid species (2n = 8x = 56) showed substantial DNA variability. Densitometric and flow-cytometric analyses clearly showed that many interphase nuclei are beyond the expected 2C-4C limit characteristic of cycling cells. The most distinct DNA instabilities were observed in B. carinatus, where we found especially high numerical chromosome variations and many chromosome numbers higher and lower than 2n = 8x = 56. Such quantitative DNA variability is substantially reduced beyond the meristematic part of the roots and virtually absent within leaf mesophyll cells. Estimated nuclear 2C DNA content was 11.63 pg in B. arvensis, 23.03 pg in B. hordeaceus, 22.94 pg in B. carinatus, 12.99 pg in B. willdenowii, 24.65 pg in B. erectus and 24.54 pg in B. inermis. By DNA amount, the analyzed taxa formed two distinct groups: with 2C DNA value at about 12 pg (B. arvensis and B. willdenowii), and at about 24 pg (B. hordeaceus, B. carinatus, B. erectus and B. inermis). Estimated DNA values were not linked with ploidy level. All species showed a similar, mainly telomeric heterochromatin distribution, but small amounts of intercalary and centromeric heterochromatin were also seen. The majority of analyzed species also had similar heterochromatin amounts within karyotype. The only exception was B. carinatus with 16.20% heterochromatin (calculated as percent of karyotype length). It is suggested that the observed 77% nuclear DNA difference between two closely related representatives of subg. Ceratochloa (B. carinatus and B. willdenowii) is to some extent a result of heterochromatin accumulation in B. carinatus chromosomes.

Key words: Bromus L., chromosome number, karyotype, heterochromatin, DNA amount, flow cytometry, C-banding.

Disorders of Microsporogenesis in Transgenic Tobacco Plants with Altered Proportions of Native Histone H1 Variants

 Joanna Ślusarczyk1, Marta Prymakowska-Bosak2, Marcin Przewłoka2, Andrzej Jerzmanowski2,3 and Mieczysław Kuraś4

            1 Department of Ecology and Environmental Protection, Świętokrzyska Academyul.Świętokrzyska 15, 25-343 Kielce, Poland
Laboratory of Plant Molecular Biology, Warsaw University
, ul. Pawińskiego 5a,02-106 Warsaw, Poland
Institute of Biochemistry and Biophysics, Polish Academy of Sciences
,  ul. Pawińskiego 5a,02-106 Warsaw, Poland
Department of Plant Morphogenesis, Warsaw University
, ul. Miecznikowa 1,02-096 Warsaw, Poland

 Received April 18, 2001; accepted July 19, 2001

The role of histone H1 in the regulation of flower growth and development and in microsporogenesis was investigated in previously obtained transgenic tobacco plants expressing the antisense histone H1 cDNA. These plants exhibited a characteristic male-sterility phenotype caused by altered proportions of the native histone H1 in chromatin. The type and frequency of chromosomal aberrations during male meiosis as well as the distortions affecting tetrad formation in transgenic plants are reported.

Key words: Tobacco, histone H1, meiosis, chromosomal aberrations, male sterility.

Holokinetic Chromosomes of Luzula luzuloides (Juncaceae) in Callus Culture

Agata Madej and Elżbieta Kuta*

Department of Plant Cytology and Embryology, Jagiellonian University, ul. Grodzka 52, 31-044 Cracow, Poland

 Received January 15, 2001; accepted April 4, 2001

A long-term callus culture from Luzula luzuloides leaf meristem subcultured for over one year was examined cytologically. In the control material most of the mitotic cells (95.97%) represented diploid level and standard chromosomes in terms of length (2n = 12AL). Aneuploidy occurred with low frequency (4.03%), with somatic chromosome numbers 2n = 13, 14 resulting from partial agmatoploidy. Karyotype analysis of control material showed differences in chromosome length ranging from 4.94 um to 3.19 um in prometaphase, 3.54 um to 2.54 um in mid metaphase, and 2.81 um to 1.88 um in late metaphase. Callus cells exhibited a wide range of chromosome number variation (2n = 7-48), although a high percentage of cells (61.39%) represented the standard karyotype (2n = 12AL). Variability in chromosome number and karyotype structure was a consequence of chromosome fission (partial and total agmatoploidy), chromosome fusion (partial symploidy) as well as aneuploidy and polyploidy. There was no evident correlation between the frequency of structural and numerical chromosome variation and the duration of callus culture. The cells with modified karyotype appeared in particular collections.

Key words: Luzula luzuloides, holokinetic chromosomes, callus culture, chromosome variation, agmatoploidy, symploidy, aneuploidy, polyploidy.


Interaction: Immunocytochemical Studies on Petunia Hybrida Hort.

 Marta Lenartowska1*, Maria Isabel RodríGuez-GarcíA3 and Elżbieta Bednarska2

 1Laboratory of Developmental Biology, Institute of General and Molecular Biology Nicolaus Copernicus University, 87-100 Toruń, Poland
Academy of Education, Institute of Biology and Environmental Protection
, 76-200 Słupsk, Poland
Department of Biochemistry, Cell and Molecular Biology of Plants Estación Experimental del Zaidín (CSIC), E-18008 Granada, Spain

 Received June 7, 2001; accepted August 2, 2001

A commercial anti-calmodulin monoclonal antibody (CaM MAb) was used to determine the presence and localization of calmodulin (CaM) and calmodulin-like protein in unpollinated and pollinated styles of Petunia hybrida. In the unpollinated style, CaM was localized in the cytoplasm of transmitting tract cells. The response to pollination and/or pollen tube growth was the presence of bound CaM-like protein in the extracellular matrix (ECM) of transmitting tissue. Gold particles were localized in the ECM surrounding the tips of the growing pollen tubes. In the pollen tubes, CaM epitopes were present mainly in and around some vesicles of the apical cytoplasm. The results suggest that CaM and CaM-like protein are involved in pollen tube growth in vivo.

Key words: Petunia hybrida, calmodulin, calmodulin-like protein, extracellular matrix, pollen-pistil interaction, pollen tube, style.

Pollen and Seed Morphology of Diploids and Natural Tetraploids of Trifolium pratense L. (Leguminosae)

N. Münevver Pinar* , H. Nurhan Büyükkartal and Hatice Çölgeçen

Department of Biology, University of Ankara, 06100 Ankara, Turkey

 Received November 26, 2000; accepted April 30, 2001

Pollen and seed morphology were examined in diploids (2n = 14) and natural tetraploids (2n = 28) of Trifolium pratense L. The pollen morphology was generally correlated with ploidy level. Great variety in pollen size and apertures was noted in the tetraploid form. Exine structure was studied by TEM. Seed size, color and coat surface ornamentation can be distinguished between the diploid and tetraploid forms.

Key words: Trifolium pratense L., Leguminosae,  pollen, seed.


Isolation and Antimitotic Activity of Polysaccharides from Fruit Bodies of Xerocomus badius (Fr.) Kühn. Ex Gilb.

Janina Węgiel1, Grażyna Końska1, Jean Guillot2, Bożena Muszyńska1, Jacques Bohatier2 and Stanisław Kohlmünzer1

 1Department of Pharmaceutical Botany, Collegium Medicum, Jagiellonian University, ul. Medyczna 9, 30-688 Cracow, Poland
Groupe de Recherche en Biologie Cellulaire et Environnement Universite d'Auvergne - Clermont 1, Faculte de Pharmacie 28 pl. H. Dunant, F- 63000
Clermont-Ferrand, France

Received February 22, 2001; accepted June 18, 2001

 Two polysaccharide fractions were isolated from Xerocomus badius fruit bodies with a dry weight yield of 0.41% (Polysaccharide fraction A) and 0.89% (Polysaccharide fraction B). Their sugar content was 99.5% and 98.7%, respectively. Polysaccharide fraction A was composed of glucose, while Polysaccharide fraction B contained mainly glucose and mannose. Biological activity testing using the Allium test showed mitostatic and mitodepressant activity for both fractions. Polysaccharide fraction A exhibited a mitostatic action at a concentration of 0.5%, while Polysaccharide fraction B showed such activity at 0.5% and 0.25% concentrations after 24 h and 48 h. Polysaccharide fraction B exerted a direct influence on cell division of the tested organism in the Tetrahymena test.

 Key words: Xerocomus badius, glucan, glucomannan, Allium test, mitotic index, Tetrahymena test.

Apoplastic system of the gynoecium and embryo Sac in relation to function

 Hong-yuan Yang

 Key laboratory of MOE for Plant Developmental Biology, Wuhan University, Wuhan 430072, China

Received March 22, 2001; accepted June 8, 2001

This article addresses the gynoecium and embryo sac from a fresh viewpoint, i.e. the apoplastic system involved in the pistil and their putative functions in sexual processes. This system includes the extracellular matrix of the pollen tube track with particular emphasis on the micropyle and synergids, the intercellular space between the cells of the female germ unit, and the apoplast surrounding the embryo sac. Most of the data cited are research results during the last decade, mainly from ultrastructural and cytochemical observations, that give some interesting and important information about the apoplastic system concerned.

Key words: Apoplast, extracellular matrix, gynoecium, pollen tube track, micropyle, synergid, embryo sac, calcium.


Organogenesis Initiation and Plant Regeneration from Hypocotyls and Cotyledons of Androgenic Embryos of Brassica napus L.

   Agnieszka Bagniewska1 , Teresa Cegielska-Taras2 and Maciej Zenkteler1

 1 Laboratory of General Botany, Institute of Experimental Biology, Adam Mickiewicz University, Al. Niepodległości 14, 61-713 Poznań, Poland
Laboratory of Plant Tissue Culture, Plant Breeding and Acclimatization Institute
, ul. Strzeszyńska 36, Poznań, Poland

 Received February 19, 2001; accepted May 14, 2001

Regeneration of plants from hypocotyls and cotyledons of androgenic embryos of winter oilseed rape (Brassica napus L. homozygous line DH-O 120) were investigated. Both types of explants were cultured on Murashige and Skoog (MS) and Gamborg (B5) media with or without a low concentration of growth regulators added. Buds showed the highest developmental ability on B5 medium in the presence of 0.5 mg 1-1GA3. All shoots and secondary embryos derived from cotyledons and hypocotyls were of cortical origin. Starch grains gradually disappeared as shoot initiation proceeded, indicating their role in the process of organogenesis.

 Key words: Brassica napus, haploid, androgenic embryos, plant regeneration.

   webmaster: Piotr Osyczka