Acta Biologica Cracoviensia s. Botanica

Abstract Distribution of genetic diversity among and within plant populations may depend on the mating system and the mechanisms underlying the efficiency of pollen and seed dispersal. In self-incompatible species, negative frequency-dependent selection acting on the self-incompatibility locus is expected to decrease intensity of spatial genetic structure (SGS) and to reduce population differentiation. We investigated two populations (peripheral and more central) of wild service tree (Sorbus torminalis (L.) Crantz), a self-incompatible, scattered tree species to test the differences in population differentiation and spatial genetic structure assessed at the self-incompatibility locus and neutral nuclear microsatellites. Although, both populations exhibited similar levels of genetic diversity regardless of the marker type, significant differentiation was noticed. Differences between FST and RST suggested that in the case of microsatellites both mutations and drift were responsible for the observed differentiation level, but in the case of the S-RNase locus drift played a major role. Microsatellites indicated a similar and significant level of spatial genetic structure in both populations; however, at the S-RNase locus significant spatial genetic structure was found only in the fragmented population located at the north-eastern species range limits. Differences in SGS between the populations detected at the self-incompatibility locus were attributed mainly to the differences in fragmentation and population history.

Abstract Studies concerning the ultrastructure of the periendothelial zone integumentary cells of Asteraceae species are scarce. The aim was to check whether and/or what kinds of integument modifications occur in Onopordum acanthium. Ovule structure was investigated using light microscopy, scanning electron microscopy, transmission electron microscopy and histochemistry. For visualization of calcium oxalate crystals, the polarizing microscopy was used. The periendothelial zone of integument in O. acanthium is well developed and composed of mucilage cells near the integumentary tapetum and large, highly vacuolated cells at the chalaza and therefore they differ from other integumentary cells. The cells of this zone lack starch and protein bodies. Periendothelial zone cells do not have calcium oxalate crystals, in contrast to other integument cells. The disintegration of periendothelial zone cells was observed in a mature ovule. The general ovule structure of O. acanthium is similar to other members of the subfamily Carduoideae, although it is different to “Taraxacum”, “Galinsoga” and “Ratibida” ovule types.

Abstract The purpose of this study was to illuminate the effects of fulvic acid in plants’ stress signaling pathway. 2.0 mg/l fulvic acid was sprayed on soybean leaves for 3 days at 12 h intervals, followed by treatment of 150 mM NaCl or exposed to heat stress at 35°C for 2 h over 2 days. Pre-treatment with fulvic acid increased the relative water content (RWC), antioxidant enzyme, isoenzyme activities (SOD, APX, GST), as well as alleviated the stress-induced oxidative damage by decreasing the levels of hydrogen peroxide (H2O2) and malondialdehyde (MDA). In addition, the application of fulvic acid under salt stress induced rubisco expression only at 12 h, while it induced the expression of cytochrome c oxidase at 6 h and 12 h. On the other hand, fulvic acid under heat stress induced significant expression of both rubisco and cytochrome c oxidase at 6 h and 12 h. However, under high salinity conditions, fulvic acid suppressed the transcript levels of Hsp70, while it induced increases in Hsp70 levels under heat treatment at 6 h. As a result, in this study, fulvic acid played the role of a regulator and stimulant in stress response of soybean leaves.

Abstract An efficient micropropagation protocol for production of genetically uniform clones of Eryngium campestre L. was developed. To determine the effect of nutritional and hormonal factors on shoot and root development and bioactive compounds production, three variants of media differing in the content of macro- and micronutrients, as well as plant growth regulators of various types and concentrations were tested. The highest regeneration (100%), with over 13 shoots per explant, was induced on Murashige and Skoog (MS) medium with 1.0 mg l−1 benzyladenine (BA) and 0.1 mg l−1 indole-3-acetic acid (IAA). The in vitro derived shoots multiplied through axillary bud formation were rooted and transferred to an experimental plot with 78% frequency of survival. Flow cytometry showed no variation in nuclear DNA between the seedlings and micropropagated plants. Preliminary thin layer chromatography (TLC) analysis indicated that phenolic acids, saponins, flavonoids and acetylenes were present in plant biomass. Ultra high performance liquid chromatography (UHPLC) analysis revealed that shoots and roots from in vitro derived plants and root cultures maintained the ability to produce rosmarinic acid (RA), rosmarinic acid hexoside (RA-HEX) and chlorogenic acid (CGA). The highest phenolic acid content was detected in roots of in vitro regenerated plants. The extract from those roots expressed the highest inhibitory effect against bacteria Staphylococcus aureus, as well as dermatophytes Trichophyton mentagrophytes and T. rubrum.

Abstract Rehmannia glutinosa hairy roots were used to evaluate the effect of methyl jasmonate (MeJa) and salicylic acid (SA) on increase of root biomass and production of iridoids (catalpol, harpagide) and phenylethanoids (verbascoside and isoverbascoside). The elicitors were added to 23-day-old culture separately at concentrations between 50 and 200 μM or in combinations at concentrations of 50 and 100 μM. Roots were harvested 72 h and 120 h after elicitation. The type of elicitor, its concentration and exposure time were found to strongly affect the content of each analyzed compound. A 72-hour treatment with 200 μM MeJa was the most effective in increase of verbascoside content (60.07 mg·DW−1 equivalent to 845.45 mg·L−1) and isoverbascoside (1.77 mg·DW−1 equivalent to 24.94 mg·L−1): these respective amounts were roughly 10- and 6.4-fold higher than the control values (unelicited roots). Exposure to 150 μM MeJa provided optimal harpagide content after 72 hours (0.136 mg·DW−1; 7.5-fold increase compared to the control), and catalpol content after 120 hours (up to 2.145 mg·DW−1). The combination of MeJa and SA also resulted in higher levels of secondary metabolites compared to the control culture, although these levels were lower than those observed for MeJa alone at the optimal concentration and exposure time. SA alone was less efficient in enhancing metabolite production than MeJa.

Abstract The objective of the study was to assess the influence of methyl jasmonate (MJ) vapors on accumulation of 2-phenylethylamine (PEA), phenylacetic acid (PAA) and 2-phenylethanol (PE) in leaves and roots of maize (Zea mays L. subsp. mays, saccharata group, cv. Złota Karłowa) seedlings. Furthermore, we analyzed the expression patterns of eight genes (ADH1, ADH2, AO2, CAO, PDC1, PDC2, PTA and LOX, encoding alcohol dehydrogenase 1 and 2, primary amine oxidase, aldehyde oxidase 2, phenylalanine decarboxylase 1 and 2, phenylalanine (histidine) transaminase and lipoxygenase, respectively) involved in biosynthesis and turnover of PEA in maize tissues. In addition, the effect of MJ application on fresh biomass and growth of the tested seedlings was recorded. One-day MJ exposure increased the fresh weight of aerial parts and roots of Z. mays seedlings, whereas the opposite tendency occurred after 4-day of MJ treatment. One-day application of MJ resulted in an increase in the length of roots and its fluctuations in the aerial parts of maize plants, but extended exposure declined the growth of both parts of the seedlings. Methyl jasmonate elicitation caused various changes in the contents of PEA, PAA and PE in the maize seedlings. MJ treatments led to high upregulation of most genes, with the exception of three genes (i.e., ADH1, ADH2 and AO2) whose expression was downregulated after a 4-day exposure.

Abstract Apple species and cultivars differ in nuclear (2C) DNA content and ploidy level. The majority of these genotypes are diploids, but there are some triploids and a few tetraploids. Nuclear DNA content is a specific feature and its flow cytometric evaluation can be helpful in differentiating taxa. For many apple genotypes – including all the Polish ones, these characteristics are not known. 2C DNA was evaluated in relation to leaf, flower, fruit, pollen grain and stomata sizes as well as to the flowering time for seventy genotypes (including 46 Polish cultivars) gathered in the gene bank of the Research Institute of Horticulture, Skierniewice, Poland. For standard cultivars with the known chromosome number, 2C value was 1.71 pg for diploid cultivar ‘Alwa’ (2n=2x=34), 2.55 pg for triploid ‘Boskoop’ (3x=51), and 3.37 pg for tetraploid genome (4x=68) of mixoploid ‘McIntosh 2x+4x’. In 61 cultivars (including 41 Polish ones), the nuclear DNA content ranged from 1.58 to 1.78 pg indicating their diploid chromosome number. Five cultivars were identified as triploids (‘Bursztówka Polska’, ‘Pagacz’, ‘Rapa Zielona’, ‘Rarytas Śląski’ and ‘Witos’) owing to their nuclear DNA amount ranging between 2.42 and 2.58 pg. Leaf, flower, fruit, stomata and pollen grain sizes were on average significantly larger in triploids. Thus, in 3x plants the mean leaf surface was 49.1 cm2, flower diameter – 52.4 mm, fruit weight – 204.7 g, stomata length – 32.1 μm and pollen grain diameter – 33.7 μm, whereas in diploids – 36.0 cm2, 46.1 mm, 162.7 g, 28.4 μm and 30.7 μm, respectively. Pollen grain viability was on average significantly higher in diploids (75.6%), compared to triploids (22%). These results confirm that in apple, as in many other plant species, the higher ploidy level of triploids is generally associated with increased sizes of pollen grains, stomata, flowers, fruits and leaves but decreased pollen viability. No clear correlation between ploidy level and flowering time was found. In the case of mixoploid apple genotypes possessing diploid and tetraploid genomes, some phenotype observation is helpful in describing the ploidy level of the histogenic layers, L1 and L2. Small stomata sizes (similar to diploid) indicate diploid L1 and larger leaf sizes, compared to diploid counterparts, show tetraploid L2. The results will be used for breeding, in which it is important to determine maternal and paternal genotypes as well as the direction of the crossing that is of great importance in obtaining seeds and materials for further selection.

Abstract The present study was conducted to determine the effect of the D genome on embryoid induction and green plant regeneration in wheat anther culture and how it is influenced by low temperature and mannitol treatment. For this reason, the anther culture response of two Canadian bread wheat cultivars and their extracted tetraploids (AABB) was studied. As controls two cultivars well responding to anther-culture (i.e. cvs. Kavkaz/Cgn and Acheron) and a no-responding cultivar (cv. Vergina) were used. Approximately 3000 anthers of these cultivars were cultured and three pre-treatments were applied: cold pre-treatment for 7 and 18 days at 4°C, and 0.3M mannitol for seven days at 4°C. W14 and 190-2 were used as induction and regeneration media, respectively, and the basic MS medium as the rooting medium. No green plants were produced from the tetraploids, which supports the view that the D-genome chromosomes are necessary for androgenic response in wheat. Furthermore, the Canadian cultivars performed better after 18-day pre-treatment at 4°C. The extracted tetraploids produced fewer embryoids and performed better after seven days of cold pre-treatment. The controls well responding to anther culture performed better than the Canadian cultivars, although their best response was recorded after seven-day cold pre-treatment. Cultivar Vergina produced no green plants. The presence of mannitol influenced negatively both embryoid and green plant production. It was concluded that the D genome plays a crucial role in anther culture response of wheat and that this response is influenced by both the genotype and the duration of cold pre-treatment.

Abstract The total soluble sugar content and antioxidant enzyme activities were studied for the first time during axillary shoot formation in Magnolia × ‘Spectrum’ in vitro in response to BAP (0.3 mg l−1), different levels of gibberellic acid (GA3; 0.0, 0.1, 0.5, 1.0 mg l−1), sucrose (20 and 30 g l−1) and nitrogen salts (KNO3/NH4NO3; 100/100% and 75/50% relative to MS medium). Among various GA3 and sucrose/nitrogen salts ratios, the most effective axillary multiplication (5.9 shoots/explant) and leaf formation (25.7 leaves per multiplied clumps) were obtained after addition of GA3 at 0.1 mg l−1 to a BAP medium containing 20 g l−1 sucrose and reduced levels of nitrogen salts (75% KNO3 and 50% NH4NO3). The addition of GA3 to the BAP medium enhanced shoot formation by 36% and leaf formation by 27%. The highest shoot formation capacity of M. × ‘Spectrum’ in vitro coincided with enhanced levels of soluble sugar and peroxidase (POD) activity. Increasing GA3 concentration from 0.1 to 1.0 mg l−1 in the above medium resulted in inhibition of shoot and leaf formation and a decrease in the soluble sugar content. The influence of GA3 on the activities of catalase (CAT) and POD depended on its concentration and the levels of sucrose and nitrogen salts in the medium. The highest increase in CAT and POD activities, that coincided with the enhanced shoot formation capacity of M. × ‘Spectrum’ in vitro, was observed after addition of GA3 to the medium containing high levels of sucrose and nitrogen salts.

Abstract Leaves and internodes from Stevia rebaudiana Bertoni plants growing in different conditions were used for transformation with two strains of Agrobacterium rhizogenes: ATCC 15384 and LBA 9402. Hairy roots formation was observed and the percentage of the transformed explants depended on the type of explant, time of inoculation and inoculum concentration. Inoculation of explants from ex vitro and in vitro plants with LBA 9402 strain led to higher efficiency of transformation than inoculation with ATCC 15384 strain. Growth rate of hairy roots in liquid culture was assessed under light and dark conditions. It was found that the growth of hairy roots decreased significantly under light conditions. Transformation of hairy roots growing in different culture conditions was confirmed at the molecular level using PCR method with primers constructed against rolB and rolC genes from A. rhizogenes.

Abstract Chromosome numbers for 12 Hieracium s.str. species from Bulgaria, Macedonia, Montenegro, Poland, Romania and Serbia are given and their metaphase plates are illustrated. Chromosome numbers are published for the first time for H. albinum Fries 2n=4x=36, H. bukovicae Rohlena & Zahn; 2n=3x=27, H. retyezatense subsp. subatratiforme Neičeff & Zahn 2n=4x=36, H. velenovskyi Freyn 2n=3x=27, as well as two undescribed species belonging to the H. bohatschianum agg. 2n=4x=36 and H. nigrescens agg. 2n=4x=36 and a hybrid between H. naegelianum Pančić and H. bifidum s.lat. 2n=4x=36.