Salt stress causes severe reduction in the growth and yield of rice plants. The ability to maintain cellular ion homeostasis is of importance to help the plant survive under salt stress. Salt overly sensitive 1 (SOS1), a plasma membrane Na+/H+ antiporter, has been proven to play critical roles in Na+ exclusion out of the cell, hence contributing to salt tolerance in plants. In this study, we analyzed the natural nucleotide polymorphisms occuring within the entire coding sequence as well as the upstream region of the OsSOS1 gene by comparing the sequences of two contrasting rice genotypes, namely, Nipponbare (salt-sensitive) and Pokkali (salt-resistant). In total, six nucleotide polymorphisms were identified in the coding sequence, and 44 nucleotide substitutions, 225-bp-insertion and 65-bp-deletion were observed in the upstream region of the OsSOS1 gene. Futher in silico analysis revealed that two out of six nucleotide polymorphisms in the coding sequence were non-synonymous (A1600G, G2204A) which led to two amino acid substitutions (T534A, S735N, respectively) positioned in the C-terminal domain of OsSOS1 transporter, but caused no effect on protein properties. In the upstream region of OsSOS1 gene, 44 single nucleotide polymorphisms and two INDELs were identified, in which nucleotide substitutions at position -1392, -1389, -822, -583, +57 and an insertion at position -1035 caused change in cis-regulatory elements. Analysis of OsSOS1 expression revealed that salt induced the expression of the gene in the roots, but not in the leaves in both investigated rice cultivars.
Abstract The fern Platycerium bifurcatum is a valuable component of the flora of tropical forests, where degradation of local ecosystems and changes in lighting conditions occur due to the increasing anthropogenic pressure. In ferns, phytochrome mechanism responsible for the response to changes in the value of R/FR differs from the mechanism observed in spermatophytes. This study analyzed the course of ontogenesis of nest leaves in P. bifurcatum at two values of the R/FR ratio, corresponding to shadow conditions (low R/FR) and intense insolation (high R/FR). The work used only non-destructive research analysis, such as measurements of reflectance of radiation from the leaves, their blue-green and red fluorescence, and the chlorophyll a fluorescence kinetics. This allowed tracing the development and aging processes in the same leaves. Nest leaves are characterized by short, intense growth and rapid senescence. The study identified four stages of development of the studied leaves related to morphological and anatomical structure and changing photochemical efficiency of PSII. Under the high R/FR ratio, the rate of ontogenesis of the leaf lamina was much slower than under the low R/FR value. As shown, the rapid aging of the leaves was correlated with faster decline of the chlorophyll content. It was shown that leaf senescence was accompanied by accumulation of polyphenols, anthocyanins and carotenoids on the basis of reflectance and fluorescence measurements in the blue-green range.
Several parasitic plants are known to have acquired mitochondrial genes via a horizontal transfer from their hosts. However, mitochondrial gene transfer in this direction has not yet been found in the parasite-rich family Orobanchaceae. Based on a phylogenetic analysis of the mitochondrial atp6 gene in selected species of Orobanche s.l., we provide evidence of a host-to-parasite transfer of this gene in O. coerulescens, which is a Eurasiatic species that parasitises Artemisia (Asteraceae). We did not find the original Orobanche atp6 gene in this species, which suggests that it has been replaced by a gene that was acquired from Asteraceae. In addition, our data suggest the occurrence of a second HGT event in the atp6 sequence – from Asteraceae to Phelipanche. Our results support the view that the transfer of genetic material from hosts to parasites influences the mitochondrial genome evolution in the latter.
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.
This paper is both a review and a study. It discusses the taxonomic status of Yellow Archangel (Galeobdolon luteum Huds.) from historical and contemporary perspectives, and gives a comprehensive list of synonyms for the discussed genera, species and lower taxonomic units, including their publication details. In the study it is postulated that G. luteum should be included in the genus Lamium. The hypothesis is verified by a comparative analysis between the representatives of the genera Galeobdolon and Lamium in four DNA regions: ITS, accD, rpoC1 and trnH-psbA. The analysis supported the determination of phylogenetic relationships among the studied taxa: G. luteum is not genetically distant enough from Lamium to be considered a separate genus, and integration of Galeobdolon and Lamium is legitimate.
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.
Abstract The influence of sodium alginate sterilization on the viability and mitotic activity of embedded protoplasts was studied in protoplasts of Brassica oleracea subsp. alba and rubra isolated from hypocotyl tissue and leaves of seedlings or plants grown in vitro. Both leaf and hypocotyl-derived protoplasts were more viable and divided more frequently when embedded in filtrated alginate. Division frequency was highest in cv. Reball F1 and the mitotic activity of its protoplasts was three times higher when embedded in filtrated alginate (36.1 ± 6.8%) than when cultured in autoclaved alginate (10.9 ± 5.0%). Protoplast-derived calli colonies were transferred to solid regeneration media and plants of all tested accessions were obtained.
Abstract In vitro cultures of Anethum graveolens (dill) were maintained on the Linsmaier and Skoog (LS) medium – three variants, and the Murashige and Skoog (MS) medium – seven variants, which contained different amounts of plant growth regulators, cytokinin (BAP) and auxin (NAA) (from 0.1 mg l−1 to 3.0 mg l−1). Methanolic extracts from in vitro grown biomass were analyzed by HPLC for free phenolic acids and furanocoumarins. The total amounts of free phenolic acids on the LS medium variants were similar (35.23–38.65 mg 100 g−1 DW), but higher on the MS variants, ranging from about 66 mg 100 g−1 DW to 100 mg 100 g−1 DW. The main metabolites were: p-hydroxybenzoic acid (max. 24.41 mg 100 g−1 DW) on the LS−based media, and salicylic acid (max. 57.88 mg 100 g−1 DW) and p-hydroxybenzoic acid (max. 36.27 mg 100 g−1 DW) on the MS−based media. The total amounts of furanocoumarins were lower, as they did not exceed 8.5 mg 100 g−1 DW on the LS media and 25 mg 100 g−1 DW on the MS media. The main compounds in this group were bergapten (max. 15.01 mg 100 g−1 DW) and marmesin (max. 8.12 mg 100 g−1 DW). The MS variant containing 0.5 mg l−1 BAP and 2.0 mg l−1 NAA was proposed as the best production medium for both groups of metabolites. The maximum total amounts of free phenolic acids obtained in the in vitro grown biomass were slightly higher than their amounts in the fruits of the mother plant analyzed for comparison (99.66 mg 100 g−1 DW and 93.34 mg 100 g−1 DW, respectively); the maximum total amounts of furanocoumarins were approximately 1.8 times higher than in the fruits (24.26 mg 100 g−1 DW and 13.67 mg 100 g−1 DW, respectively).
Abstract The genetic diversity of potato cultivars collected from Yunnan Province was evaluated using 24 pairs of SSR markers. SSR analysis of 24 pairs of primers showed varying degrees of polymorphism among the 85 cultivars: 297 of the 304 bands were polymorphic. The primers yielded between 5 (STM2028) and 19 (StI029) bands (mean 12). The ratio of polymorphic bands ranged from 83.33% to 100% (mean 97.75%). Polymorphism information content (PIC) varied from 69.31% to 93.67% (mean 86.47%). Genetic similarity ranged from 0.5987 to 0.7632, indicating relatively low genetic diversity in the potato cultivars from Yunnan Province. Cluster analysis by UPGMA and PCA clearly delineated the genetic relationships of all cultivars; 83 of the 85 cultivars could be discriminated by only two pairs of primers, STM0030 and STM1104. The high polymorphism and good resolution of the primers used in this study make them good tools for discriminating potato cultivars.
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.
We compared the biochemical profiles of Physalis ixocarpa hairy roots transformed with Agrobacterium rhizogenes ATCC and A4 strains with non-transformed root cultures. The studied clones of A4- and ATCC-induced hairy roots differed significantly; the latter showed greater growth potential and greater ability to produce secondary metabolites (tropane alkaloids) and to biotransform hydroquinone to arbutin. We compared glucose content, alanine and aspartate aminotransferase activity, and L-phenylalanine ammonia-lyase activity. We analyzed markers of prooxidant/antioxidant homeostasis: catalase, ascorbate peroxidase, oxidase, glutathione peroxidase and transferase activity, and the levels of ascorbate, glutathione, tocopherol and lipid peroxidation. We found that transformation induced strain-specific regulation, including regulation based on redox signals, determining the rate of allocation of carbon and nitrogen resources to secondary metabolism pathways. Our results provide evidence that A. rhizogenes strain-specific modification of primary metabolites contributed to regulation of secondary metabolism and could determine the ability of P. ixocarpa hairy root clones to produce tropane alkaloids and to convert exogenously applied hydroquinone to pharmaceutically valuable arbutin. Of the studied parameters, glucose content, L-phenylalanine ammonia-lyase activity and alanine aminotransferases activity may be indicators of the secondary metabolite-producing potential of different P. ixocarpa hairy root clones.
Abstract The study investigates the genetic differentiation among two subspecies of Allium ursinum L., namely A. ursinum subsp. ursinum and subsp. ucrainicum as well as their putative hybrid that is represented by individuals with intermediate morphology. Inter-Simple Sequence Repeats (ISSR) were applied to determine the status of intermediate morphotypes in terms of their genetic pattern and to assess the level of genetic variability within and between various populations of A. ursinum. The study comprises 144 specimens from nine populations along the east-west transect in Poland, which includes localities of both subspecies and their putative hybrid. Among the examined populations, 48 bands were amplified, of which 45 were found to be polymorphic. The principal coordinate analysis (PCoA), the neighbour-net analysis and Mantel test showed a strong correlation between genetic variability and geographic distance. Analysis of molecular variance (AMOVA) revealed that a greater proportion of total genetic variation resided within populations rather than among them. The Structure Bayesian clustering analysis revealed the presence of three distinct genetic groups within studied populations, where ‘eastern’ genotypes correspond to A. ursinum subsp. ucrainicum, and ‘western’ to subsp. ursinum; whereas the third genetic group has the largest share in the individuals occurring at the border of the distribution ranges of both subspecies. The emergence of the third genetic group is probably an effect of hybridization events occurring within the secondary contact zone. Typical morphologically intermediate populations occur only in a relatively narrow geographical zone, but the hybrid zone revealed by molecular markers is actually much wider than it is suggested by the morphological pattern of individuals. The current distribution pattern of both subspecies of A. ursinum and their hybrid zone is related to the two main directions of postglacial migration of Fagus sylvatica to the area of Poland. The hybrid zone arose as an effect of the secondary contact of two divergent lineages of A. ursinum.
Abstract During greening, excised etiolated barley leaves and cucumber cotyledons that were depleted of exogenous Ca2+ by a chelating agent (ethylene glycol-bis (beta aminoethyl ether)-N,N,N`N`-tetraacetic acid, EGTA) showed ∼50% reduced chlorophyll (Chl) accumulation and ∼30% accumulation of apoprotein of the light-harvesting chlorophyll a/b-binding protein complex of photosystem II (LHCPII). The Ca2+ channel blocker lanthanum chloride (LaCl3) applied to cucumber cotyledons reduced LHCPII accumulation more than EGTA did. In both plant mate-rials, cytokinins enhanced chlorophyll accumulation by 50-60% and this effect was completely canceled by EGTA application. Hormones significantly increased LHCPII accumulation but EGTA application reduced that effect in barley leaves by ∼30% and in cucumber cotyledons by ∼80%. A similar effect was observed in LaCl3-treated cotyledons. CaCl2 application boosted chlorophyll accumulation in both plant materials. CaCl2 applied together with cytokinin reduced the hormonal effect on chlorophyll accumulation by ∼38% in barley leaves and 23% in cucumber cotyledons, but almost totally inhibited cytokinin-stimulated LHCPII accumulation. Our results indicate that calcium variously mediates the effect of cytokinin on chlorophyll and LHCPII accumulation. Cytokinin-induced enhancement of chlorophyll accumulation seems totally dependent on the exogenous pool of Ca2+, while Ca2+-dependent and Ca2+-independent pathways are involved in the hormonal effect on LHCPII accumulation. The effect of cytokinin on the increase of light-induced LHCPII accumulation appears to be sensitive to exogenously applied Ca2+, which almost totally blocked the hormonal effect. Our results give indirect evidence that the responses to cytokinin and light act on different events leading to Chl and LHCPII accumulation.
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
Abstract Cell pattern and ultrasculpture were examined by light and scanning electron microscopy in bulb tunics of 46 Allium species to determine the diagnostic value of micromorphological characters. The study examined the diversity of these characters, evaluated their usefulness at different taxonomic levels (species, section, subgenus), and considered the results in relation to the recent intrageneric classification of the genus. Detailed characteristics are provided for the investigated species, and taxa showing the presence of calcium oxalate crystals in bulb tunic cells are indicated. The results suggest that several bulb tunic characters are of taxonomic significance in Allium as their variation between specimens of the same species was negligible; they can be useful elements of species descriptions and determination keys. Allium subgenus Allium shows considerable variation of bulb tunic ultrasculpture and hexagonal or elongated cell patterns. Differences in ultrasculpture are sufficient to distinguish species within the Amerallium subgenus. Three subgenera (Anguinum, Butomissa, Reticulatobulbosa) are characterized by fibrous tunics with reticulate ultrasculpture. Rectangular to elliptic cells with thick walls, giving the bulb tunic an almost perforated structure, are characteristic for Allium subgenus Cepa. No specific pattern was found for Allium subgenus Melanocrommyum and Polyprason. The only representative of subgenus Microscordum (Allium monanthum) showed distinct herringbone ultrasculpture. The bulbs of Allium subgenus Rhizirideum representatives can be distinguished by their linear ultrasculpture following the long axis of the elongated cells. Allium bulb tunic ultrasculpture and cell pattern show some degree of variability. These characters are of potential use in taxonomic delimitation, species determination and further study of the relationships between species, particularly in members of subgenus Amerallium.
To keep genetic diversity, flowering plants have developed a self-incompatibility system, which can prevent self-pollination. It has been reported that calcium concentration in pistil papilla cells was increased after self-pollination in transformed self-incompatible Arabidopsis thaliana. In this study, we found that CML27 changed its expression level for both mRNA and protein when compared to transcriptome and proteome. At the same time, CML27 was expressed in the anther and pistil at a high level and reached up to 5-fold up-regulated expression in the pistil at 1 h post-pollination when compared to 0 min. In order to find out potential proteins that may interact with BoCML27, BoCML27 was expressed in and isolated from E. coli. After its co-incubation with Brassica oleracea pistil proteins, the products were separated on SDS-PAGE gels. We found a specific band at the position between 130–180 kDa. Through LC-MS-MS (Q-TOF) analysis, eight proteins were identified from the band. The proteins include 26S proteasome non-ATPase regulatory (26S), Phospholipase D, alpha 2 (PLDα2) involved in Ca2+ binding and Coatomer subunit alpha-2-like (Coatomer) involved in vesicle mediated transport. All of these identified proteins provide new insights for the self-incompatibility response in B. oleracea, specific for increasing Ca2+ concentration in pistil papilla cells.
Abstract NH4+ is an important N-source which regulates plant growth and development. However, the underlying mechanism of NH4+ uptake and its-mediated signaling is poorly understood. Here, we performed phosphoproteomic studies using the titanium dioxide (TiO2)-mediated phosphopeptides collection method together with LC-MS analysis. The results indicated that phosphorylation levels of 23 and 43 peptides/proteins involved in diverse aspects, including metabolism, transport and signaling pathway, were decreased and increased respectively after NH4+ treatment in rice roots. Among 23 proteins detected, IDD10, a key transcription factor in ammonium signaling, was identified to reduce phosphorylation level of S313 residue. Further biochemical analysis using IDD10-GFP transgenic plants and immunoprecipitation assay confirmed that NH4+ supply reduces IDD10 phosphorylation level. Phosphorylation of ammonium transporter 1;1 (AMT1;1) was increased upon NH4+ treatment. Interestingly, phosphorylation of T446, a rice specific residue against Arabidopsis was identified. It was also established that phosphorylation of T452 is conserved with T460 of Arabidopsis AMT1;1. Yeast complementation assay with transformation of phosphomimic forms of AMT1;1 (T446/D and T452/D) into 31019b strain revealed that phosphorylation at T446 and T452 residues abolished AMT1;1 activity, while their plasma membrane localization was not changed. Our analyses show that many proteins were phosphorylated or dephosphorylated by NH4+ that may provide important evidence for studying ammonium uptake and its mediated signaling by which rice growth and development are regulated.
This study used ISSR markers to assess the genetic diversity of a collection of 15 genotypes of Salix purpurea and 6 interspecific hybrids, employing 40 of 60 tested ISSR primers generating polymorphic amplification products. The PCR-ISSR method was adapted for S. purpurea by optimizing the annealing temperature for each primer. The polymorphism index of ISSR amplification products was 91.8% for all studied genotypes and 70.4% for S. purpurea genotypes. Nei's genetic identity statistics ranged from 0.538 to 0.958. Nei's genetic distance values were used to build a dendrogram (UPGMA) for the investigated genotypes. The dendrogram shows five clusters, and principal coordinate analysis yielded nearly the same genetic relationships among the studied genotypes. The results confirm the usefulness of ISSR markers for determining genetic diversity in S. purpurea.
Genetic diversity is often considered a major determinant of long term population persistence and its potential to adapt to variable environmental conditions. The ability of populations to maintain their genetic diversity across generations seems to be a major prerequisite for their sustainability, which is particularly important for keystone forest tree species. However, little is known about genetic consequences of demographic alterations occurring during natural processes of ecological succession involving changes in the species composition. Using microsatellites, we investigated genetic diversity of adult and offspring generations in beech (Fagus sylvatica L.) and oak (Quercus robur L.) populations coexisting in a naturally established old-growth forest stand, showing some symptoms of ongoing ecological succession from oak- to beech- dominated forest. In general, adult generations of both species exhibited high levels of genetic diversity (0.657 for beech; 0.821 for oak), which, however, depended on the sets of selected genetic markers. Nevertheless, several symptoms such as differences in genetic diversity indices between generations, significant levels of inbreeding (up to 0.029) and low estimates of effective population size (48-80) confirmed the declining status of the oak population. On the other hand, the uniform distribution of genetic diversity indices across generations, low levels of inbreeding (0.004), low genetic differentiation among adults and offspring and, most importantly, large estimates of effective population size (119-716), all supported beech as a successive and successful tree species in the studied forest stand.
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.
Abstract We aimed to produce tissue cultures and plant regeneration from endangered Crocus species: C. scepusiensis, C. tommasinianus, C. vittatus (“Verni” series of the genus) and C. banaticus. For initiation of cultures we used a plant growth regulator (PGR) combination used for in vitro culture of saffron and its relatives: 10 mg L-1 α-naphthaleneacetic acid (NAA) and 1 mg L-1 6-benzyladenine (BA). Shoot tips of young seedlings (C. scepusiensis) and corms (for the rest of species) were used as explants. C. scepusiensis explants developed into organogenic calli. On media with decreased NAA and with or without increased BA concentration, calli produced stigma-like structures and/or shoots and whole plants. In the other species, callus initiation medium induced callus formation with abundant somatic embryos. In C. tommasinianus, embryos developed shoots when auxin content of medium was decreased. In C. banaticus, a decrease of auxin with or without an increase in cytokinin content led to shoot or whole plant regeneration, as in C. scepusiensis. In the case of C. vittatus and C. banaticus, initiation and/or maintenance of cultures on indole-3-butyric acid (IBA) and increased sucrose concentration stimulated whole plant regeneration and in vitro cormlet development. C. scepusiensis and the rest of cultures (organogenic vs. embryogenic) differed at the biochemical level: C. scepusiensis cultures had higher (yet still low) enzymatic antioxidant (catalase, peroxidase) activities. With respect to catalase isoenzyme patterns, C. banaticus was different from the rest of cultures, demonstrating its distinct taxonomical position. Besides germplasm preservation use of the present cultures, they have a potential biotechnological value.