We studied the embryology of Withania somnifera (L.) Dunal by light microscopy in order to reveal specific embryological features of the genus, and compared the results with embryological data on other members of the family Solanaceae. The key embryological characters of W. somnifera include dicotyledonous-type anther wall formation, simultaneous cytokinesis in pollen mother cells, binucleate tapetal cells, 2-celled mature pollen, anatropous, tenuinucellate and unitegmic ovules, polygonum-type embryo sac formation, the presence of an endothelium, and cellular endosperm formation. We give the first report of the dicotyledonous mode of anther wall formation (previously described as basic type) for the species. Comparative study suggests that anther wall formation, number of nuclei in tapetal cells, number of cells in mature pollen, mode of embryo sac formation and endosperm development are the most variable embryological features in Solanaceae. Some of these embryological features of W. somnifera should be of value for comparative study of related species and their phylogenetic relationships within the family.

SDS-PAGE electrophoresis was used to study the effect of NaCl on protein expression in two cultivars of tomato (Solanum lycopersicum L.): Edkawi (salt-tolerant) and Castle rock (salt-sensitive). Five-day-old seedlings were grown on MS agar media supplemented with 0, 50, 100, 150, 200 and 300 mM NaCl. Two days after treatment the seedlings were examined to determine the effect of salt on their growth and to relate that to protein banding variations. Gel analysis showed differences in at least 4 protein bands with molecular weights at 20, 25, 45 and 65 kDa. These proteins were induced in the 50 mM NaCl treatment in the salt-sensitive cultivar, then decreasing to undetectability at higher concentrations. In the salt-tolerant cultivar, most of the proteins exhibited a more or less steady expression pattern and maintained expression through the 200 mM NaCl treatment. All proteins gave weak or no expression signals at 300 mM NaCl, the treatment that proved lethal. Differentially expressed bands were identified using MALDI-TOF mass spectrometry. The putative function of each identified protein in relation to salt stress is discussed.

We examined the response of plants of various crop and weed species to cyanamide in order to evaluate allelochemical- mediated interactions between the species. We studied germination and seedling growth in the common weeds Galium aparine L. and Amaranthus retroflexus L., and the crops Zea mays L., Triticum aestivum L., Lactuca sativa L., Solanum lycopersicum L. and Sinapis alba L. as acceptor plants. Concentration-dependent phytotoxic effects of cyanamide were noted during seed germination and in the root and shoot growth of the tested plants. The monocotyledonous plants generally were less sensitive to cyanamide treatment. Seed germination and seedling growth of the dicotyledonous plants were strongly inhibited by the allelochemical at both tested concentrations (1.2 mM, 3 mM). We conclude that cyanamide has potential for use as a natural herbicide only in specific field systems of cyanamide-tolerant monocotyledonous crops accompanied by cyanamide-sensitive dicotyledonous weeds.

We analyzed DNA damage, mitotic activity and polyploidization in Crepis capillaris callus cells during short- and long-term in vitro culture, and the influence of plant growth regulators on these processes. Changes in the concentration of growth regulators altered the stability of callus. The level of DNA damage was highly dependent on the growth regulator composition of the medium. Cytokinin at high concentrations damaged DNA in the absence of auxin. Short- and long-term callus differed in sensitivity to growth regulators. Mitotic activity changed when callus was transferred to medium with modified growth regulators. Callus cell nuclear DNA content increased with age and in response to plant growth regulators. Hormones played a role in the genetic changes in C. capillaris callus culture. We demonstrated the usefulness of C. capillaris callus culture as a model for analyzing the effect of culture conditions, including plant growth regulators, on genetic stability.

We studied the effect of qualitative and quantitative variation of saponin content in foliar tissues of four European alfalfa (Medicago sativa L.) cultivars (Radius, Sapko, Sitel, Radius line 1) on pea aphid (Acyrthosiphon pisum Harris) development, and the effect of aphid infestation on alfalfa saponin content. Aphids (adult apterae, larvae, and adult alatae) were counted on 3-, 6- and 9-month-old plants (before the first, second and third cutting). Thin-layer chromatography was used to detect and estimate the quantity of the following saponins: 3GlcA, 28AraRhaXyl medicagenic acid; 3Glc, 23Ara, 28AraRhaXylApi zanhic acid (zanhic acid tridesmoside); and 3RhaGalGlcA soyasapogenol B (soyasaponin I). Radius, Sapko, and Sitel contained all three saponins but Radius line 1 did not contain zanhic acid tridesmoside or medicagenic acid glycoside. Saponin content was highest in Radius and lowest in Radius line 1. Regardless of the cultivar, saponin content was higher in aphid-infested than uninfested plants. For all sampling dates, aphid numbers were highest on Radius line 1 and lowest on Radius; that is, aphid numbers were inversely related to saponin content. Alfalfa has a herbivoreinduced defense. Saponin levels increase in the foliage of infested alfalfa. Attempts of plant breeders to reduce saponin content in order to increase alfalfa digestibility for livestock might make the plants more susceptible to aphids and other pests.

Chromosome numbers for 15 taxa of Hieracium s.lat. (including two taxa of Pilosella Vaill.) from Bosnia and Herzegovina, Bulgaria, Greece, North Macedonia, Poland, Romania and Slovakia are given and their metaphase plates are illustrated. Chromosome numbers are published for the first time for H. pannosum subsp. parnassi Nägeli & Peter from Greece (3x and 4x), and for an undescribed species of H. sect. Cernua from North Macedonia (4x). A new, diploid chromosome number was found in H. bracteolatum s.lat. from Greece.

The influence of ambient solar UV-A or UV-B radiation on growth responses was investigated in three varieties of cotton (Gossypium hirsutum L.) after exclusion of solar UV-A/B radiation: JK-35, IH-63 and Khandwa-2. Cotton plants were grown from seeds in UV-exclusion chambers lined with selective UV filters to exclude either UV-B (280-315 nm) or UV-A/B (280-400 nm) from the solar spectrum under field conditions. Excluding UV-B and UV-A/B significantly increased plant height, leaf area and dry weight accumulation in all three varieties of cotton. The varieties differed considerably in their sensitivity to ambient UV-A/B. Khandwa-2 was most sensitive and JK-35 least sensitive to ambient solar UV. We monitored the activity of the antioxidant enzymes superoxide dismutase (SOD), ascorbic acid peroxidase (APX), glutathione reductase (GR) and guaiacol peroxidase (GPX), as well as the level of the antioxidant ascorbic acid (ASA), in primary leaves of the most UV-sensitive variety (Khandwa-2). The level of UV-B-absorbing substances was significantly decreased by exclusion of solar UV-B and UV-A/B. Exclusion of solar UV decreased the activity of all the antioxidant enzymes monitored and the level of ascorbic acid versus control plants (+UV-A/B) grown under filters transparent to solar UV. Reduction of the antioxidant defense after UV exclusion indicates that ambient solar UV exerts significant stress and induces some reactive oxygen species to accumulate, which in turn retards the growth and development of cotton plants. Ambient solar UV stresses cotton plants, shifting their metabolism towards defense against solar UV. Exclusion of solar UV eliminates the need for that defense and leads to enhancement of primary metabolism.

The free-living Acanthamoeba sp. causes various diseases. Treatment of them is very difficult and not always effective because of encystation, making it highly resistant to antiamoebic drugs. Gram-positive bacteria Staphylococcus aureus, Gram-negative bacteria Escherichia coli, and an yeast Candida albicans also exhibit outstanding resistance to antimicrobial substances. The search for new natural amoebicidal and antimicrobial agents of plant origin is still of current interest. The aim of the study was to investigate the amoebicidal activity of the extracts obtained from tissue culture and a field-grown plant of Chaenomeles japonica against pathogenic trophozoites of Acanthamoeba spp. and antimicrobial effect against S. aureus, E. coli, and C. albicans. The extracts of C. japonica had an inhibitory effect on the proliferation of Acanthamoeba trophozoites as compared to the non-treated control. Among the crude extracts tested, the extract of leaves, from both shoot culture and the field-grown plant had remarkable amoebicidal action against the trophozoites but also antibacterial activity against Gram-positive bacteria Staphylococcus aureus. The extract from leaves from shoot culture, already on the second and third days of treatment, showed an antiamoebicidal effect at a concentration of 1 mg mL-1 (inhibition of trophozoites 87.5% and 91.8%, respectively). In addition to leaves from shoot culture (a conc. 5 mg mL-1, 2nd day inhibition of trophozoites 85.7% and 3rd day 97.2%), leaves from a field-grown plant (a conc. 5 mg mL-1, 2nd day 91.0% and 3rd day 94.4%) and callus (a conc. 5 mg mL-1, 2nd day 90.0% and 3rd day – 95.4%) also exhibited a good antiamoebicidal activity. Out of the four extracts, the extracts from leaves from both shoot culture and a field-grown plant were reported to be the most active against Gram-positive S. aureus, which was determined by the values of MIC = 5.0 mg mL-1 and MIC = 2.5 mg mL-1, respectively. The inhibitory potential depends on the yield and composition of mainly bioactive compounds: pentacyclic terpenoids (mainly betulinic, ursolic, and oleanolic acids) and polyphenols (mainly chlorogenic acid and its isomers, epicatechin, dimeric, and trimeric proanthocyanidins, quercetin and kaempferol derivatives).

Since plant responses to selenium nanoparticles (nSe) had not been clarified, this study was carried out to evaluate the effects of nSe (10 and 100 μM) on photosynthesis performance, ion homeostasis, antioxidant system, and phenylpropanoids in strawberry exposed to salt stress. Inductively Coupled Plasma-Mass Spectroscopy analyses indicated that foliar-applied nSe can be taken up by leaves and trans-located to roots. Salinity led to an increase in Na concentration and reductions in Ca and K contents which were relieved by the nSe applications. Moreover, the nSe treatment at 10 μM alleviated the NaCl-induced lesion to PSII functioning, contributing to improvement in water-splitting complex (Fv/Fo) under salinity. The exposure to nSe at a concentration of 100 µM exhibited a moderate stress, determined by the increases in hydrogen peroxide (H2O2) and lipid peroxidation rate (membrane integrity index). The nSe10 treatment increased catalase activity and phenylpropanoid derivatives contents (salicylic acid, catechin, and caffeic acid) and decreased the content of oxidants under salinity condition. Consequently, nSe utilization at a suitable dose can be an effective method to alleviate signs of salt stress via improvements in photosynthesis, ion hemostasis, photosynthesis performance, salicylic acid (a vital signaling defensive hormone), and antioxidant machinery.

This study investigated the quantitative anatomy of photosynthetic tissues (leaf mesostructure) of wild ginseng Panax ginseng C.A. Mey. (Araliaceae) plants from different natural habitats. The structural and functional traits of the photosynthetic apparatus shown to be especially elastic were mesophyll cell volume (Cv >40%) and traits related to filling of the leaf with cells and plastids (Cv≥21%). P. ginseng possesses relatively few cells per leaf area (44.6-107.2 103/cm2) and chloroplasts (1.7-4.9 106/cm2). Also low are the values of such integral indexes as relative surface of mesophyll cells (Ames/A, 2.78-5.28) and relative surface of chloroplasts (Achl/A, 1.9-3.2). The leaf mesostructure of wild ginseng shows traits of a plant typically found in shady forest habitats. The photosynthetic apparatus of ginseng adapts to various habitat conditions on the level of leaf mesostructure, through structural transformations of mesophyll tissue, such as changes in the number and size of cells and chloroplasts and also the integral surface indexes Achl/A and Ames/A.

The spontaneous diploidization rates in oilseed rape (Brassica napus L.) via in vitro androgenesis are too low for practical applications. In contrast, artificial doubling of chromosomes of the microspore has proven to be more successful and allows homozygous plants to be obtained in a short time. Here, we present the efficiency of diploidization of B. napus haploids using three different chromosome doubling methods.

Using the in vitro approach in microspores, the rate of chromosome doubling in 24 populations of androgenic plants ranged from 15.8% to 94.0%. An alternative in vivo method for the induction of chromosome doubling involves colchicine treatment of young haploid plants, and this yielded doubling rates ranging from 47.5% to 86.4% in 10 different plant populations. Another in vivo method of chromosome doubling is colchicine treatment of the excised young axillary shoots of haploid plants at the early flowering stage. The high efficiency of this method was confirmed in haploid plant populations from 11 genetically distinct donors in which the frequency of occurrence of diploids ranged from 53.3% to 100%. However, in this case, the time required for seed formation from doubled haploids increased by about 3–5 months. The availability of several methods of chromosome doubling at various stages of the androgenic process – from isolated microspores through to young plants and flowering plants – allows seeds to be obtained from nearly every selected individual haploid.

The flowers of Polemonium caeruleum are protandrous. The nectary is in the form of a rim encircling the ovary. Secreted nectar accumulates in a chamber located at the bottom of the floral tube and is protected by dense staminal hairs. The nectary tissue is not vascularized, but is supplied by vascular strands that occur near the base of the nectary and which directly supply the stamens. Nectar is secreted via modified stomata located on the upper part of the rim, particularly on the adaxial surface. The number of stomata and the volume and sugar concentration of nectar are greater during the female stage than during the male stage. In both stages, however, the nectar is sucrose-dominant. This paper shows that in P. caeruleum the nectar sugars are not a direct product of current photosynthesis, since plastids of nectary cells are devoid of chlorophyll. The main source of sugars in secreted nectar is the phloem sap, together with starch that accumulates in the nectary cells during the male stage and is then rapidly hydrolyzed during the female stage.

The chemical composition and bioactivity of a water/methanol extract prepared from aerial parts of Circaea lutetiana were determined. HPLC-DAD-MS3 analysis revealed the presence of 14 different compounds comprising phenolic acids, ellagitannins and flavonoids. Antioxidant assays showed the extract's strong scavenging activity towards DPPH (SC50 33.1±3.1 μg/ml), O2 - (SC50 4.0±2.3 μg/ml) and H2O2 (SC50 below 2 μg/ml). Enzyme-based studies revealed that Circaea lutetiana extract inhibits the activity of hyaluronidase (IC50 13.3±2.4 μg/ml) and lipoxygenase (IC50 44.7±1.4 μg/ml). These results support some traditional uses of Circaea lutetiana.

The effects of gamma irradiation on the vernalization requirements, growth and development of winter wheat grown in a rainout shelter were studied during two successive growing seasons. Dry grains of winter wheat cv. Kobra were irradiated with 300 Gy radiation from a cobalt 60 gamma irradiator. Treated and control grains were pregerminated and subjected to vernalization for 0, 42 or 54 days. Morphological parameters of the plants developing from irradiated seeds (M1 generation) and the plants grown from the seeds produced by the irradiated plants (M2 generation) were measured in order to track the studied effects over two generations. Irradiation of dry grains slowed the growth and development of the plants regardless of the temperature treatment. The measured yield structure elements appeared to be lower for irradiated plants, but no clear effect of radiation on vernalization requirements was noted

Achene macro- and micromorphology and pericarp anatomy are described in four Polish species of Anemone (A. narcissiflora, A. nemorosa, A. ranunculoides, A. sylvestris). Biometric analysis showed that achene size varies greatly in all the studied species and is of limited diagnostic value. Three types of sculpture connected with the character of the indumentum were distinguished. The presence or absence of stomata on the achene style and the character of the hair base differentiated A. nemorosa and A. ranunculoides, which have the same type of pericarp ornamentation. The endocarp (number of layers and outline of its cells) was shown to be useful in the systematics of Anemone

The Arabidopsis CDKG;2 gene encodes a putative cyclin-dependent Ser/Thr protein kinase of unknown biological function. This gene shows structural similarity to animal and human cyclin-dependent (PITSLRE) kinases. This study used the homozygous knockout cdkg;2 mutant based on T-DNA insertional line SALK_090262 to study the effect of mutation of the CDKG;2 gene on explant response and in vitro plant regeneration. For callus induction and proliferation, hypocotyls and cotyledons of 3-day-old seedlings of cdkg;2 and A. thaliana ecotype Col-0 were cultured on solid MS medium supplemented with 2,4-D (2 mg l-1). Organogenesis was induced after callus transfer on MS + TDZ (0.5 mg l-1). The initiation time of callus and shoot induction differed between the mutant and control cultures. Shoot regeneration after callus transfer on MS + TDZ was delayed in cdkg;2 (31 days versus 7 days in Col- 0). Shoots formed on callus derived from Col-0 hypocotyls but not on cotyledon-derived callus; in cdkg;2, shoots developed on both callus types. Mutant shoots did not form roots, regenerants were dwarfed, and inflorescences had small bud-like flowers with a reduced corolla and generative organs. Abnormalities observed during cdkg;2 organogenesis suggest a role of CDKG;2 as a regulator of adventitious root initiation

Using four Polish Vicia faba L. minor cultivars (Bronto, Dino, Tibo, Nadwiślański) we obtained callus from epicotyl fragments collected from 7- and 14-day-old seedlings and from cotyledonary nodes of immature seeds. Callus induction efficiency varied from 81% to 97% depending on the origin of the explant. Shoots regenerated only from the cotyledonary nodes of all tested cultivars. On average, 50% of the explants grown on MS medium containing 1.0 mg dm-3 NAA, 0.5 mg dm-3 BAP, 0.25 mg dm-3 GA3, 1.0 g dm-3 casein hydrolysate, 750 mg dm-3 inositol, 3% sucrose and 0.4% agar were able to regenerate shoots. The number of calluses regenerating shoots was highest from explants collected from fruiting nodes 6 to 9. Decapitation of donor plants increased the percentage of calluses regenerating shoots. On half-strength MS medium with 2 mg dm-3 NAA and on 1/2 MS alone, 11% of the shoots rooted; on 1/2 MS with 1 g dm-3 AC, 8.0% rooted. The regenerants were transferred to Perlite with Hoagland medium and acclimated. Ten percent of the regenerated plants survived the acclimation process, flowered and produced seeds.

Metallothioneins are low-molecular-weight proteins capable of covalently binding heavy metal ions due to the presence of many cysteine residues in their sequences. We analyzed the predicted amino acid sequences of 19 metallothionein (7 from Arabidopsis thaliana and 12 from Oryza sativa) and their promoter sequences in silico in order to determine the potential regulatory cis-elements present in the promoters of metallothionein genes, from which it is possible to determine the putative functions of these genes. The PlantCARE and PLACE databases provided information about the putative regulatory elements in the metallothionein promoters. Metal response element sequences were found in the promoters of eleven O. sativa and two Arabidopsis metallothionein genes. Copper response elements were identified in both model plants, usually in many copies, particularly in O. sativa. Both the high cysteine content and the presence of metal response motifs in the promoters support the suggestion that metallothioneins play a key role in metal detoxification. The most common putative element in the analyzed promoters was CIRCADIAN, which was present in five A. thaliana and eight O. sativa sequences. The methyl jasmonate response sequence, root-specific expression element and drought response element were found only in O. sativa metallothioneins. Light and low temperature response elements, biotic and abiotic stress elements, an abscisic acid-responsive element and an ethylene-responsive element occur in selected metallothionein promoters of both species. A few promoters have putative organ- and cell-specific regulatory elements. The presence of many different motifs in the promoters of the Arabidopsis and O. sativa genes implies that metallothioneins are general stress response proteins with many important functions in plants, including regulation of their normal development and adaptation to changing environmental conditions.

We used germination tests to assess the frequency of polyembryony in 9 asparagus cultivars with a high propensity to produce double embryos with different ploidy levels: Alpha, Andreas, Boonlim, Cipres, Eposs, Helios, Limbras, Ravel and Sartaguda. Twin embryos inside a single seed were found in 3 cultivars: Eposs 2n, Ravel 2n and Sartaguda 2n, at 0.60% frequency (15 seeds with twin embryos out of 2500 seeds). Of 30 obtained seedlings, 14 were separated diploid-diploid twins, 6 were conjoined diploid pairs, 8 were separated diploid-haploid and 2 were diploid-haploid pairs conjoined in the hypocotyl region. Some embryos showed unilateral dominance of one embryo (size and shape). The haploid status of the smallest embryo was confirmed by chromosome number (n=x=10) and flow cytometry (nuclear C DNA amount 1.95 pg). The haploid obtained in this manner possessed enough vegetative vigor to undergo chromosome doubling.

Here we report the consequences of telomere erosion in Arabidopsis thaliana, studied by examining seed and pollen production and the course of male meiosis through the last five generations (G5-G9) of telomerase-deficient Arabidopsis mutants. We used a previously described mutant line in which telomerase activity was abolished by T-DNA insertion into the TERT gene encoding telomerase reverse transcriptase. Reduced fertility accompanied by morphological abnormalities occurred in G6, which produced on average 35 seeds per silique (vs. 43 in wild type) and worsened in G7 (30 seeds) and G8 (14 seeds), as did the morphological abnormalities. The last generation of tert mutants (G9) did not form reproductive organs. Analysis of meiosis indicated that the main cause of reduced fertility in the late generation tert mutants of Arabidopsis was the numerous chromosomal end-to-end fusions which led to massive genome rearrangements in meiocytes. Fusion of meiotic chromosomes began in G5 and increased in each of the next generations. Unpaired chromosomes (univalents) were observed in G7 and G8. The study highlights some differences in the meiotic consequences of telomere shortening between plant and animal systems.

Hladnikia pastinacifolia RCHB., a narrow endemic, has an extremely restricted distribution in Trnovski gozd (Slovenia), despite the presence of many sites with suitable habitats. We compared the morphological traits of plants from different populations and habitats. The overall pattern showed that the smallest plants, with low fruit number, are found on Èaven (locus classicus or type locality); the largest individuals, with high fruit number, grow in the Golobnica gorge. As judged by plant size and seed set, the optimal habitats are screes. We used RAPD markers to estimate genetic variation between and within populations, as well as between and within the northern and the southern parts of the distribution area. Hladnikia showed only a low level of RAPD variability. AMOVA partitioned the majority of genetic diversity within selected populations. The low genetic differentiation between populations and their genetic depauperation indicates survival in situ, since the Trnovski gozd plateau most likely was a nunatak region in the southern Prealps during Pleistocene glaciations. Later range expansion of extant populations was limited by poor seed dispersal. We also analyzed the cpDNA trnL-F intergenic spacer to check whether the sequence is useful for studying the phylogenetic relationships of Hladnikia within the family Apiaceae (Umbelliferae). Our results support the assertion that H. pastinacifolia is an old taxon.

Nanotechnology has been widely applied in agriculture, and understanding of the mechanisms of plant interaction with nanoparticles (NPs) as environmental contaminants is important. The aim of this study was to determine the effects of foliar application of cobalt oxide (Co3O4) NPs on some morpho-physiological and biochemical changes of canola (Brassica napus L.) leaves. Seeds were sown in plastic pots and grown under controlled conditions. Fourteen-day-old seedlings were sprayed with different concentrations of Co3O4 NPs (0, 50, 100, 250, 500, 1000, 2000, and 4000 mg L-1) at weekly intervals for 5 weeks. Growth parameters of the shoot (length, fresh and dry weights) were stimulated by low concentrations of Co3O4 NPs (50 and 100 mg L-1) and repressed by higher concentrations. Similar trends were observed in photosynthetic pigment contents. The results indicated that high concentrations of Co3O4 NPs increased lipoxygenase (LOX) activity and the malondialdehyde (MDA), hydrogen peroxide (H2O2), and dehydroascorbate (DHA) contents, but reduced the membrane stability index (MSI), ascorbate (ASC), and glutathione (GSH). Despite the increase of antioxidant capacity (DPPH) and the accumulation of nonenzymatic antioxidants (total flavonoids and flavonols) and osmolytes (proline, glycine betaine (GB) and soluble sugars) at high concentrations of Co3O4 NPs, the growth and photosynthesis were reduced. The defence system activity did not seem to be sufficient to detoxify reactive oxygen species (ROS). Altogether, high concentrations of Co3O4 NPs showed a phytotoxic potential for canola as an oilseed crop.

Plant viruses create many changes in the morphology of the plant cell once the infection process has begun. This paper describes and compares the ultrastructural changes induced in maize cells by two isolates of Maize dwarfmosaic virus (MDMV), Spanish (MDMV-Sp) and Polish (MDMV-P), and one isolate of Sugarcane mosaic virus (SCMV) at 10 and 42 days post-inoculation: the concentration and arrangement of virus particles, inclusion bodies associated with infection, and other cytological alterations. The most important difference between maize cells infected with MDMV isolates and with SCMV-P1 was in the form of cytoplasmic cylindrical inclusions. In cells infected with MDMV only typical inclusions such as pinwheels and scrolls were observed, but laminar aggregates were also present in SCMV-infected cells. No virus particles were found in plant cell organelles. Specific virion arrangements occurred in cells infected with MDMV-Sp and SCMV. The most interesting new finding was of specific amorphous inclusions in the cytoplasm of MDMV-Sp-infected cells, which clearly differentiated the two MDMV isolates studied.