Intraspecific changes in genome size and chromosome number lead to divergence and species evolution. Heavy metals disturb the cell cycle and cause mutations. Areas contaminated by heavy metals (metalliferous sites) are places where microevolutionary processes accelerate: very often only a few generations are enough for a new genotype to arise. This study, which continues our long-term research on Viola tricolor (Violaceae), a species occurring on both metalliferous (Zn, Pb, Cd, Cu) and non-metalliferous soils in Western and Central Europe, is aimed at determining the influence of environments polluted with heavy metals on genome size and karyological variability. The genome size of V. tricolor ranged from 3.801 to 4.203 pg, but the differences between metallicolous and non-metallicolous populations were not statistically significant. Altered chromosome numbers were significantly more frequent in material from the polluted sites than from the non-polluted sites (43% versus 28%). Besides the standard chromosome number (2n = 26), aneuploid cells with lower (2n = 18-25) or higher (2n = 27, 28) chromosome numbers were found in plants from both types of site, but polyploid (2n = 42) cells were observed only in plants from the metalliferous locality. The lack of correlation between chromosome variability in root meristematic cells and genome size estimated from peduncle cells can be attributed to elimination of somatic mutations in generative meristem, producing chromosome-stable non-meristematic tissues in the peduncle.
Cucumber (Cucumis sativus L. cv. Dar) leaves exposed to UV-B irradiation at a biologically effective dose of 9.5 kJ m-2d-1 showed decreased chlorophyll fluorescence parameter values versus the control; in peppermint (Mentha piperita L. cv. Asia) leaves those values were almost unchanged after treatment. Fv/Fo and Rfd were reduced more than other values, indicating inhibition of the oxygen-evolving complex and cooperation between the light and dark photosynthesis reactions as the primary targets of UV-B. The photosynthetic electron transport rate showed less change directly after irradiation, but after 24 h of recovery it was reduced to 50% of the control. Generally, photosystem II of peppermint leaves appeared more tolerant to the applied UV-B radiation than in cucumber leaves.
We investigated the antioxidant defense mechanism, metal uptake and lipid peroxidation (LPO) levels at different leaf positions in Mentha piperita L. grown in Mn2+-deficient and control conditions. Under manganese deficiency the activity of superoxide dismutase (SOD), catalase (CAT) and guaiacol peroxidase (GuaPOX) and the content of ascorbate, chlorophyll, and carotenoid under Mn2+ deficiency were significantly lower than in the control for all leaf positions. SOD activity correlated positively with Mn2+ uptake. Fe2+ uptake was inhibited by Mn2+ deficiency. During early stages of Mn2+ deficiency, M. piperita leaves showed relatively more antioxidant activity and lower LPO. Towards the final stages of the treatment period, comparatively lower SOD, CAT and GuaPOX activity and higher LPO levels accelerated the senescence process.
We investigated direct and indirect formation of somatic embryogenesis in Brassica oleracea var. botrytis (cauliflower), a very important vegetable crop worldwide. Direct somatic embryogenesis, which is rather rare, was achieved in culture of 2-week-old hypocotyl explants of Brassica oleracea var. botrytis on MS medium supplemented with 1.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5; 1.0; and 1.5 mg/l kinetin. Initial induction of embryogenic callus was achieved on MS supplemented with very low concentrations of 2,4-D (0.05 mg/l and 0.1 mg/l). Indirect somatic embryogenesis from leaf sections was obtained on MS supplemented with 0.05 or 0.1 mg/l 2,4-D. We examined various stages of somatic embryos (globular, heart, torpedo, cotyledonary). More embryos per explant were produced through the indirect pathway (23-25) than through the direct pathway (14-19). The number of embryos produced was high. There is a potential for recurrent, repeated or secondary somatic embryogenesis, possibly an unlimited source for mass propagation and ideal for synthetic seed production in this species. Plant regeneration was achieved on half-strength MS medium without any hormones.
We examined whether allelochemical stress leads to increased lipoxygenase activity in roots of sweet maize (Zea mays L. ssp. saccharata), pea (Pisum sativum L.) and radish (Raphanus sativum L. var. radicula). The lipoxygenase activity of soluble and membrane-bound fractions was assessed in roots after exposure to ferulic and p-coumaric acids. Lipid peroxidation and membrane injury were determined as indicators of stress. Increased lipoxygenase activity of both studied fractions was followed by lipid peroxidation and plasma membrane injury. The results suggest the key role of lipoxygenase in plasma membrane injury during allelochemical stress caused by administration of hydroxycinnamic acids.
We used via light and scanning electron microscopy to study the leaf epidermis of five Solidago taxa from south-western Poland. Light microscopy was employed to describe the epidermal surface, including stomatal types, the shape of epidermal cell walls, stomatal density, the distribution of stomata between the abaxial and adaxial epidermis, and stomatal guard cell length. From these observations we calculated the stomatal index (SI) and stomatal ratio (SR) as the basis for defining the type of leaf. From LM of transverse sections of leaf we described mesophyll structure, the presence of secretory canals, adaxial and abaxial epidermis thickness, and leaf thickness. We examined cuticular ornamentation, trichome features and epicuticular secretions by SEM. As determined by discriminatory analysis, the most important traits distinguishing these taxa were the stomatal index of the adaxial epidermis, leaf thickness, features of the walls of epidermal cells, and the presence and features of trichomes. On the basis of observations and measurements we created a key for distinguishing Solidago taxa.
In order to evaluate morphological and physiological traits related to drought tolerance and to determine the best criteria for screening and identification of drought-tolerant genotypes, we grew two tolerant genotypes (MCC392, MCC877) and two sensitive genotypes (MCC68, MCC448) of chickpea under drought stress (25% field capacity) and control (100% field capacity) conditions and assessed the effect of drought stress on growth, water relations, photosynthesis, chlorophyll fluorescence and chlorophyll content in the seedling, early flowering and podding stages. Drought stress significantly decreased shoot dry weight, CO2 assimilation rate (A), transpiration rate (E), and Psii photochemical efficiency (Fv/Fm) in all genotypes. In the seedling and podding stages, Psii photochemical efficiency was higher in tolerant genotypes than in sensitive genotypes under drought stress. Water use efficiency (WUE) and CO2 assimilation rate were also higher in tolerant than in sensitive genotypes in all investigated stages under drought stress. Our results indicated that water use efficiency, A and Fv/Fm can be useful markers in studies of tolerance to drought stress and in screening adapted cultivars of chickpea under drought stress.
Powdery mildew, caused by Blumeria graminis f. sp. tritici, is one of the most important foliar diseases of cereals. Infection by this pathogen on triticale has intensified in Poland in the last few years. In this study we examined resistance to powdery mildew in triticale hybrids possessing resistance genes Pm4b and Pm6 introduced from common wheat. The materials tested were hybrids derived from triticale crosses with common wheat cultivars carrying the desired resistance genes. The presence of the transferred genes was reflected in increased field resistance and shown by the use of molecular markers. The paper discusses the potential introduction of the genes to improve powdery mildew resistance.
Secretory ducts and cavities of roots and rhizomes are typical features of the Cardueae tribe in the Asteraceae family. We used light microscopy to analyze the anatomy of the subterranean organs of 21 species of 13 genera of the Cardueae, with particular attention to the secretory system, interpreted in taxonomic terms. The anatomy of secretory ducts varied greatly. A new measurement quotient, C1 [length of epithelial cells (longitudinal section)] and C2 [length of adjacent cells (longitudinal section)] was established. Different types of ducts are described based on type of development and the size ratios among epithelial cells. Detailed anatomical descriptions of the ducts are given, together with their occurrence in particular taxa. The simultaneous presence of various secretory ducts within a single species and their spatial position relative to other prominent anatomical features provide valuable characters for discriminating the studied Cardueae species. These analyses are of particular interest for identification of herbal drugs as, besides chemical analytical techniques such as chromatographic fingerprinting, light microscopy is a common method for purity controls and thus required in official pharmacopeias.
We examined whether peroxidase activity in cutting bases and leaves and starch content in cutting bases can be used as rooting phase markers in the elepidote rhododendron cv. ‘Babites Baltais’ (Rhododendron L.). Changes in peroxidase activity in cutting leaves and bases, as well as starch content in cutting bases, were determined in relation to anatomical stages of rhizogenesis in leaf bud cuttings treated with 1% indole-3-butyric acid (IBA+) or without IBA (IBA-). The pattern of change of peroxidase activity was similar in cutting bases and leaves of IBA- leaf bud cuttings. Three phases of adventitious root formation were identified: induction, initiation and expression. During the induction phase peroxidase activity decreased, but no anatomical changes were observed in the cuttings. Peroxidase activity increased in the initiation phase when adventitious root initials were formed. Peroxidase activity decreased during the expression phase when adventitious root primordia developed. The starch content of IBA- leaf bud cuttings decreased during the first few days and then gradually rose to maximum, followed by a sharp reduction and another increase at the end of the experiment. The changes of starch content did not coincide with rooting phases as peroxidase activity did, and cannot be used as a rooting phase marker in rhododendrons. Adventitious root formation did not occur in IBA+ leaf bud cuttings, so distinct rooting phases could not be observed. There was a significant correlation between peroxidase activity in cutting bases and leaves of IBA- leaf bud cuttings. Peroxidase activity in leaves of rhododendron leaf bud cuttings are potentially useful as a marker for rooting phases, but that requires further anatomical and physiological study of rooting in leaf bud cuttings.
Shoot tips excised from shoot culture of Salvia officinalis were encapsulated in 2% or 3% (w/v) sodium alginate and exposed to 50 mM calcium chloride for complexation. Immediately or after 6, 12 or 24 weeks of storage at 4°C, the synthetic seeds were cultured for 6 weeks on half-strength MS medium supplemented with indole-3-acetic acid (IAA) (0.1 mg/l) and solidified with 0.7% agar. The frequency of shoot and root emergence from encapsulated shoot tips was affected by the concentrations of sodium alginate and additives in the gel matrix (sucrose, gibberellic acid, MS nutrient medium) as well as duration of storage. The frequency of shoot and root induction of non-stored synthetic seeds was highest with shoot tips encapsulated with 2% sodium alginate containing 1.5% sucrose and 0.5 mg/l gibberellic acid (GA3). Shoot tips maintained their viability and ability to develop shoots even after 24 weeks of storage when they were encapsulated in 3% alginate with 1/3 MS medium, sucrose (1.5%) and GA3 (0.25 mg/l). Root formation tended to decrease with storage time. Overall, 90% of the plantlets derived from stored and non-stored synthetic seeds survived in the greenhouse and grew to phenotypically normal plants. This procedure can enable the use of synthetic seed technology for germplasm conservation of S. officinalis, a plant species of high medical and commercial value.
The paper reports meiotic studies on 50 populations comprising 12 species belonging to 5 genera of Caryophyllaceae from the Western Himalayas. The chromosome numbers in Arenaria kashmirica (n=20), Silene conoidea (n=20), S. edgeworthii (n=12 and n=24), S. moorcroftiana (n=24), S. nepalensis (n=12), Stellaria media (n=13), S. monosperma (n=13) and S. semivestita (n=13) are reported for the first time. The chromosome numbers in Lychnis coronaria (n=12) and Silene vulgaris (n=24) are given for the first time from India, along with Gypsophilla ceratioides (n=15) from the Western Himalayas. The course of meiosis varies from normal to abnormal in different populations of Silene conoidea, S. edgeworthii, S. vulgaris, Stellaria media, S. monosperma and S. semivestita. The course of meiosis was abnormal in all studied populations of Lychnis coronaria. Abnormal microsporogenesis (cytomixis, chromosomal stickiness, unoriented bivalents, formation of laggards and bridges) led to reduced pollen fertility and differences in pollen grain size.
Poor seed set is a limiting factor in alfalfa breeding, as it slows the selection response. One strategy used to overcome this problem is to search for mutations of inflorescence morphology. Long-peduncle (lp), branched-raceme (br) and top-flowering (tf) inflorescence mutations increase the number of flowers per inflorescence, but they do not improve seed set per flower. Here we assessed pollen tube growth in styles of those inflorescence mutants and we observed embryo and endosperm development in seeds 1 to 16 days after pollination (DAP). The number of pollen tubes penetrating the style and the ovary was similar in all tested mutants and in the reference cultivar Radius. At 2 DAP, fertilized ovules were 2.7-3.9 times less numerous in certain inflorescence mutants than in the short-raceme cv. Radius. Ovule degeneration progressed at 2-4 DAP in all analyzed plants. Most ovules were not properly developed in the control cultivar (62%), nor in the forms with mutated inflorescence morphology (69-86%). The number of seeds per pod was lowest in the tf form despite its having the highest number of ovules per ovary. It appears that the number of ovules per pistil is not a crucial factor in seed set in alfalfa when fertilization efficiency is very low. Both poor fertilization and gradual ovule degeneration were factors causing poor seed set in the investigated alfalfa genotypes.
Chromosome numbers of 46 Hieracium L. and Pilosella Vaill. taxa from Austria, Bulgaria, Czech Republic, Macedonia, Montenegro, Poland, Romania, Serbia and Slovakia are presented. Chromosomes numbers are given for the first time for Hieracium amphigenum Briq. 2n = 3× = 27, H. bohatschianum Zahn 2n = 4× = 36, H. borbasii R. Uechtr. 2n = 4× = 36, H. cernuum Friv. 2n = 2× = 18, H. hazslinszkyi Pax 2n = 3× = 27, H. mirekii Szeląg 2n = 4× = 36, H. polyphyllobasis (Nyár. & Zahn) Szeląg 2n = 3× = 27, H. porphyriticum A. Kern. 2n = 4× = 36, H. racemosum Waldst. & Kit. ex Willd. subsp. racemosum 2n = 3× = 27, H. scardicum Borm. & Zahn 2n = 4× = 36, H. sparsum subsp. ipekanum Rech. fil. & Zahn 2n = 4× = 36, H. sparsum subsp. peristeriense Behr & Zahn, H. sparsum subsp. squarrosobracchiatum Behr & al. 2n = 3× = 27, H. tomosense Simk. 2n = 4× = 36, H. tubulare Nyár. 2n = 4× = 36, H. werneri Szeląg 2n = 3× = 27 and Pilosella fusca subsp. subpedunculata (Zahn) Szeląg, as well as five species of Hieracium sect. Cernua R. Uechtr. not described to date and a hybrid between H. bifidum s. lat. and H. pojoritense Woł
Triploid viviparous onions [Allium x cornutum Clementi ex Visiani 1842, syn Allium cepa L. var. viviparum Metzg. (Alef.), auct.] (2n = 3x = 24), are known in some countries only as rare relict crops. In other parts of the world they are still traditionally or even commercially cultivated. In previous cytogenetic studies of the Croatian triploid viviparous onion Ljutika, Giemsa C-banding, chromosome pairing analysis during meiosis, and genomic hybridization in situ indicated a complex hybrid with highly heterozygous karyotype structure, with possible triparental genome organization. This study continues an analysis of the karyotype structure of Ljutika. Staining with fluorochromes CMA3 (Chromomycin A3) and Dapi (4,6-diamidino-2-phenylindole) confirmed previous results from Giemsa C-banding and revealed GC-rich heterochromatic regions associated mainly with chromosome ends and nucleolus organizing regions (NORs), and only a few interstitial bands. Fish mapping of the ribosomal 18S-5.8S-26S genes revealed two major rDNA signals on the short arms of two subtelocentric satellite chromosomes in almost all metaphase plates of Ljutika. The largest subtelocentric chromosome lacked rDNA signals. A significantly smaller rDNA signal was occasionally located on one small submetacentric chromosome. These results are in agreement with previously published results from identification of NORs by silver-staining technique, which confirmed a maximum three nucleoli in interphase nuclei. We discuss the molecular mechanisms underlying rearrangements and activity of ribosomal genes in the triploid karyotype.
We used chromosome data to verify the taxonomic affiliation of specimens previously recognized as Brachyactis ciliata. All analyzed plants were diploids based on x = 7 (2n = 2x = 14), the basic number characteristic for Symphyotrichum ciliatum, allowing the examined species to be shifted from the genus Brachyactis to the genus Symphyotrichum sect. Conyzopsis. The chromosome number (2n = 2x = 14) for specimens of S. ciliatum from Poland is reported for the first time.
This communication reports detection of somaclonal variation among tissue culture-raised plants of Amorphophallus rivieri Durieu, an economically important crop in China, with high content of glucomannan in its corms. A population of regenerated plants was obtained from a single donor plant of A. rivieri via corm organogenesis, and 28 plants were randomly selected as a representative sample and subjected to analysis of somaclonal variation using inter-simple sequence repeat (ISSR) markers. Of the 26 ISSR primers screened, 13 gave distinct and reproducible band patterns, yielding 131 bands with an average of 10.1 bands per primer. Ten primers were polymorphic and generated 16 polymorphic bands with 12.2% mean polymorphism. Based on the ISSR data from the regenerated plants and the donor plant, Jaccard's similarity coefficients were calculated; they ranged from 0.961 to 1.000 with a mean of 0.982. A dendrogram was constructed using the unweighted pair group method with arithmetic mean (Upgma); it showed that a majority of regenerated plants (including the donor plant) clustered closely, with a mean similarity coefficient of 0.987. Low somaclonal variation observed in the regenerated plants indicates that rapid propagation of A. rivieri via corm organogenesis is a practicable method with a low risk of genetic instability.
We used the Dpph method to assess in vitro the antiradical activity of extracts from the roots, leaves and fruits of six Rumex L. (dock) species. Data from preliminary screening indicated that all the tested extracts showed antioxidant properties. The degree of antiradical activity depended upon the plant part. Fruit extracts from R. hydrolapathum Huds., R. obtusifolius L. and R. confertus Willd. showed stronger antiradical properties than the other tested material. We also determined tannin content levels in the extracts and their correlation with antioxidant activity.
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