Brevundimonas balnearis sp. nov., isolated from the well water of a thermal bath.

A novel alphaproteobacterium was isolated from the well water of a thermal bath at Budapest, Hungary. Phylogenetic analysis of the novel strain showed that this bacterium belongs to a distinct lineage among the genus Brevundimonas. Based on the 16S rRNA gene sequence strain FDRGB2bT showed the highest sequence similarity values to Brevundimonas naejangsanensis BIO-TAS2-2T (97.35 %), Brevundimonas viscosa F3T (97.28 %), Brevundimonas vesicularis LMG 2350T (97.27 %), Brevundimonas nasdae GTC 1043T (97.14 %), Brevundimonas vancanneytii LMG 2337T (97.13 %) and Brevundimonas aurantiaca DSM 4731T (97.13 %). The newly isolated bacterium was strictly aerobic, and its optimum growth occurred at 20-30 °C, between pH 8-9 and without NaCl. Movement was with a single polar flagellum, but the cells could also produce stalks. The major isoprenoid quinone of strain FDRGB2bT was Q-10, the major cellular fatty acids were C18 : 1ω7c and C16 : 0, and the polar lipid profile contained phosphatidylglycerol, diphosphatidylglycerol, two unknown phospholipids and four unknown glycolipids. The characteristic diamino acid in its cell wall is meso-diaminopimelic acid. The G+C content of DNA of the type strain was 69.8 mol%. Strain FDRGB2bT (=DSM 29841T=NCAIM B.02621T) is proposed as the type strain of a novel species with the proposed name Brevundimonas balnearis sp. nov.

First Report of Tomato chlorosis virus in Tomato from Hungary.

A disease of tomato (Lycopersicon esculentum Mill.) was observed in three greenhouses in Tömörkény in southern Hungary in the autumn of 2007. Thirty percent of the plants were chlorotic and stunted and had mottled leaves with interveinal yellowing and necrosis. Similar symptoms induced by Tomato chlorosis virus (ToCV) on tomato have been reported in other countries (1,2). ToCV is a Crinivirus in the Closteroviridae family, which can cause a decline in plant vigor and reduced fruit yield. ToCV is transmitted by whiteflies (Trialeurodes vaporariorum West., T. abutilonea Hald., and Bemisia tabaci Genn.) and grafting, but cannot be transmitted mechanically. Only T. vaporariorum is known to be present and widespread in Hungary. Virus presence was confirmed by reverse transcription-PCR as described by Louro et al. (2). cDNA synthesis with ToCV specific primers (ToCV-UP 5'-TCATTAAAACTCAATGGGACCGAG-3' (3) and ToCV-DW 5'-GCGACGTAAATTGAAACCC-3') was successful and electron microscopy revealed ToCV-like particles. The PCR product has been sequenced (GenBank Accession No. HQ444266) and showed 97 to 99% identity to ToCV isolates in GenBank. According to the symptoms, amplified region, sequence data, and electron microscopy, the tomato samples from Tömörkény were confirmed to be infected with ToCV. The economic losses associated with ToCV were minor. To our knowledge, this is the first report on the occurrence of ToCV in Hungary. References: (1) G. P. Accotto et al. Plant Dis. 85:1208, 2001. (2) D. Louro et al. Eur. J. Plant Pathol. 106:589, 2000 (3) J. Th. J. Verhoven et al. Plant Dis. 87:872, 2003.

Dynamics and localization of H2O2 production in elicited plant cells.

H(2)O(2) produced in plant cells plays a dual role. In addition to its antimicrobial effect, it also acts as a secondary messenger initiating and modulating responses of plants exposed to unfavorable external signals. A suspension culture of Rubia tinctorum cells challenged with elicitors was used as a model system to investigate H(2)O(2) formation. Cellular H(2)O(2) was measured by a modified titanium(IV) method, while that in the medium was detected with scopoletin fluorescence. Localization of H(2)O(2) production at the ultrastructural level was carried out by the CeCl(3) reaction. A fungal elicitor induced H(2)O(2) production with transient maxima, the first of which appeared 4 min after treatment. Three subsequent maxima appeared in the cells up to 48 h after treatment. Exposure of cells to exogenous jasmonic acid and salicylic acid also changed the H(2)O(2) concentration maxima over 48 h; however, their timing was slightly shifted. Fungal-elicitor, jasmonic acid, and salicylic acid treatments had different effects on the H(2)O(2) concentration in the medium. Ultrastructural investigations revealed that electron-dense precipitates were present at the plasmalemma and in some nearby vesicular cytoplasmic structures 30 min after treatment. Later samples showed cytochemical-precipitate accumulation in the cell walls. These deposits appeared to be local and independent of the direction of the external signal. We could not detect the presence of H(2)O(2) in peroxisomes, mitochondria, plastids, or the central vacuolar space. Electron energy loss spectroscopy investigations distinguished between the cerium-containing precipitates and other electrondense particles, thereby proving that H(2)O(2) generation occurs locally.

Disintegration of the prolamellar body structure at high concentrations of Hg2+.

The effects of high concentrations of Hg (2+) (10 (-2) M and 10 (-3) M) were investigated on the ultrastructure and on the light-induced transformation of isolated prolamellar bodies (PLBs) of dark-grown wheat leaves. Our earlier work on wheat leaf homogenates ( , Plant Biology 6, 358 - 368) showed that, depending on the concentration, Hg (2+) reacts with protochlorophyllide, NADPH and the NADPH : protochlorophyllide oxidoreductase (POR, EC 1.3.1.33) enzyme and induces disaggregation of the macrodomain structure of this latter. Spectroscopic analyses confirmed that 15 min incubation with 10 (-2) M Hg (2+) at 4 degrees Celsius completely inhibited the activity of POR also in isolated PLBs. Ultrastructural investigations revealed the loosening of the PLB structure in the Hg (2+)-treated sample, i.e., intensive vesicle formation on the surface of the PLB membranes. The hexagonal geometry of the inner lattice was not disturbed, however, the unit cell size significantly increased. The disruption of the PLB membranes upon irradiation was studied after 40 min incubation with 10 (-3) M Hg (2+) at 4 degrees Celsius and a subsequent irradiation for 40 min at 20 degrees Celsius. Equimolar concentrations (10 (-3) M) of NADPH and Hg (2+) were added to the samples 10 min prior or after the addition of Hg (2+). Our results suggest that Hg (2+) accelerates the disruption of the PLB membranes and that NADPH can only partially prevent this process. These membrane transformations were similar to those observed in the initial steps of the Shibata shift of control samples.

Rejuvenation of ageing bean leaves under the effect of low-dose stressors.

The effect of low concentrations of some stress-inducing compounds like Cd, Pb, Ni, and Ti salts and DCMU on the senescence of chloroplasts was investigated in detached primary leaves of bean. After the petioles of ageing leaves had developed roots, these low-dose stressors stimulated chlorophyll synthesis and photosynthetic activity, as compared to the control, thus causing rejuvenation in treated leaves. The amount of photosystem I (lowest in DCMU-treated leaves) and light-harvesting complex II increased, while that of photosystem II decreased or remained unchanged. Fluorescence induction parameters indicated unchanged electron transport (except for DCMU treatment). CO2 fixation and, in some cases, starch accumulation was stimulated. In parallel, the occurrence of large plastoglobuli seemed to decrease in plastids of heavy metal-treated leaves. A cytokinin bioassay of leaf extracts confirmed the cytokinin-mediated effect of low-dose stressors, as the slopes of Chl and cytokinin curves were similar during the rejuvenation process. It is assumed that these stressors generate non-specific alarm reactions, which involve changes in the hormonal balance by increasing the synthesis of cytokinins.

Tissue specific protochlorophyll(ide) forms in dark-forced shoots of grapevine (Vitis viniferaL.).

Cuttings of grapevine (Vitis vinifera L. cv. Chardonnay) were dark-forced at least three weeks. Pigment contents, 77 K fluorescence emission, excitation spectra of the leaves, petioles, stems, transmission electron micrographs of the etioplasts from leaves, the chlorenchyma tissues of the stems were analysed. The dark-grown leaves, stems contained 8 to 10, 3 to 5 mug/g fresh weight protochlorophyllide, its esters, respectively. HPLC analysis showed that the molar ratio of the unesterified, esterified pigments was 7:3 in the shoot developed in darkness. The dark-forced leaves contained carotenoids identified as: neoxanthin, violaxanthin, antheraxanthin, lutein, beta-carotene. Detailed analyses of the fluorescence spectra proved that all tissues of the dark-forced shoots had protochlorophyllide or protochlorophyll forms with emission maxima at 628, 636, 644, 655, 669 nm. The 628, 636 nm emitting forms were present in all parts of the dark-forced shoot, but dominated in the stems, which may indicate an organ specificity of the etioplast development. Variations in the distribution of the pigment forms were even found in the different tissues of the stem. The subepidermal layers were more abundant in the 655 nm form than the parenchyma cells of the inner part of the cortex, the pith. In the latter cells, the plastid differentiation stopped in intermediary stages between proplastids, etioplasts. The plastids in the subepidermal layers had developed prolamellar body structures, which were similar to those of etiolated leaves. The results highlight the importance of organ-, tissue specificity of plastid differentiation for chlorophyll biosynthesis, greening of different plant organs.

Sun and shade leaves? Cuticle ultrastructure of Jurassic Komlopteris nordenskioeldii (Nathorst) Barbacka.

An ultrastructural transmission electron microscope (TEM) study of fossil leaf cuticles from the Jurassic pteridosperm Komlopteris nordenskioeldii (Nathorst) Barbacka from the Mecsek Mountains (South Hungary) was conducted. Remnants of cuticles of leaves originating from so-called "sun and shade" environments were sectioned with a diamond knife, transversally as well as longitudinally. Although the present study showed a simple type of cuticle in this pteridosperm, differences were observed in the occurrence of its components, such as electron lucent amorphous material and various densities of granules, which give rise to different zones. The included fibrilous elements appeared to be made of aggregated and aligned granules, equivalent in size and electron density to nearby non-fibrilous granular regions. The combinations of these ultrastructural features allow distinctions between four types of cuticle: sun upper, sun lower, shade upper and shade lower. Considering the distinction made earlier in two types of cuticle and supposed to be related to sun and shade on the basis of macroscopical and microscopical features, four types only on the basis of differences in thickness, the present study reinforces the distinctions with ultrastructural microcharacteristics. As this study shows the variations in ultrastructure of cuticle among the four types, the differences observed may reveal the great sensitivity of some plants to environment. At the same time, it points out the importance, in ultrastructural studies of cuticles, of studying a number of samples for one taxon.

The Stomatal Ontogeny and Structure of the Liassic Pteridosperm Sagenopteris (Caytoniales) from Hungary.

Stomatal ontogeny is often inferred but rarely documented for extinct fossil plants because it requires observations from young leaves that are rarely preserved as fossils. The discovery of several very young leaves of the Jurassic plant Sagenopteris (Caytoniales) in the Mecsek Mountains (southern Hungary) in a good state of preservation provides the opportunity for studying the stomatal ontogenesis of this genus. The specimens show perigenous anomocytic stomata. This feature confirms the evolutionarily high position of Sagenopteris among fossil gymnosperms and supports the opinion that the ancestors of angiosperms and some groups of pteridosperms might be closely related. Such clear examples of stomatal development have not previously been documented for fossil material.

Basidiocarp and mycelium morphology of Ganoderma lucidum Karst. Strains isolated in Hungary.

Morphological, anatomical and cultural characteristics of 14 Ganoderma lucidum (Fr.) Karst strains isolated in Hungary have been investigated. Macroscopically the basidiocarps of the Hungarian strains are absolutely identical with those of described previously about the Ganoderma lucidum species-complex. Microscopic features of the fruitbodies and basidiospores showed some differences from the typical G. lucidum species. Pilocystidia, forming a homogeneous layer on the surface of the pileus, have smooth heads without protrusions and stalks not ramifying. Cell wall pillar density and width of the basidiospores also differ from that of regarded to be characteristic to G. lucidum. Although according to several authors chlamydospore formation is a characteristic feature of G. lucidum it has not been observed in mycelial cultures of the Hungarian strains. Antagonistic reactions between the Hungarian and Far Eastern G. lucidum isolates were mostly similar to the interspecific reactions between the two species G. lucidum and G. applanatum and corresponded only in a few cases to the interactions within one species. Our results suggest that the Hungarian strains significantly differ from the Far Eastern strains. To determine the taxonomic degree of this divergence genetical examinations should be carried out.