Photosynthetic Pigments of Tomato Plants under Conditions of Biotic Stress and Effects of Furostanol Glycosides

The adaptogenic effect of furostanol glycosides (FG) on biosynthesis of photosynthetic pigments in tomato plants (Lycopersicon esculentum Mill.) was studied under conditions of biotic stress caused by root-knot nematode (Meloidogyne incognita Kofoid et White). Treatment of plants with 5 × 10^–4 M FG was accompanied by an increase in the rate of biosynthesis of pigments (particularly, chlorophyll b and carotenoids), which was observed against the background of a decrease in the relative contribution of -carotene and an increase in the relative contribution of pigments of the violaxanthin cycle (VXC) to the overall pool of carotenoids. It was suggested that FG stimulated phytoimmunity by shifting metabolism of carotenoids toward enhanced biosynthesis of VXC pigments. These pigments play a protective role and facilitate stabilization of the photosynthetic apparatus, which is particularly important under stress conditions.     

Targeting molecular interactions essential for Plasmodium sexual reproduction

Malaria remains one of the most devastating infectious diseases, killing up to a million people every year. Whereas much progress has been made in understanding the life cycle of the parasite in the human host and in the mosquito vector, significant gaps of knowledge remain. Fertilization of malaria parasites, a process that takes place in the lumen of the mosquito midgut, is poorly understood and the molecular interactions (receptor–ligand) required for Plasmodium fertilization remain elusive. By use of a phage display library, we identified FG1 (Female Gamete peptide 1), a peptide that binds specifically to the surface of female Plasmodium berghei gametes. Importantly, FG1 but not a scrambled version of the peptide, strongly reduces P. berghei oocyst formation by interfering with fertilization. In addition, FG1 also inhibits P. falciparum oocyst formation suggesting that the peptide binds to a molecule on the surface of the female gamete whose structure is conserved. Identification of the molecular interactions disrupted by the FG1 peptide may lead to the development of novel malaria transmission‐blocking strategies.     

Cytokinin‐dependent specification of the functional megaspore in the Arabidopsis female gametophyte

The life cycle of higher plants alternates between the diploid sporophytic and the haploid gametophytic phases. In angiosperms, male and female gametophytes develop within the sporophyte. During female gametophyte (FG) development, a single archesporial cell enlarges and differentiates into a megaspore mother cell, which then undergoes meiosis to give rise to four megaspores. In most species of higher plants, including Arabidopsis thaliana, the megaspore closest to the chalaza develops into the functional megaspore (FM), and the remaining three megaspores degenerate. Here, we examined the role of cytokinin signaling in FG development. We characterized the FG phenotype in three triple mutants harboring non‐overlapping T–DNA insertions in cytokinin AHK receptors. We demonstrate that even the strongest mutant is not a complete null for the cytokinin receptors. Only the strongest mutant displayed a near fully penetrant disruption of FG development, and the weakest triple ahk mutant had only a modest FG phenotype. This suggests that cytokinin signaling is essential for FG development, but that only a low threshold of signaling activity is required for this function. Furthermore, we demonstrate that there is elevated cytokinin signaling localized in the chalaza of the ovule, which is enhanced by the asymmetric localization of cytokinin biosynthetic machinery and receptors. We show that an FM‐specific marker is absent in the multiple ahk ovules, suggesting that disruption of cytokinin signaling elements in Arabidopsis blocks the FM specification. Together, this study reveals a chalazal‐localized sporophytic cytokinin signal that plays an important role in FM specification in FG development.     

Interspecific Hybrids between Fusarium oxysporum lycopersici and Fusarium graminearum by Mycelial Anastomoses

Fusarium oxysporum lycopersici (FOL) and F. graminearum (FG) developed mycostatin (M) and cycloheximide (C) resistant mutants (FOL[M] and FG[C]) after treatment with nitrosoguanidine. Four hybrids, obtained from hyphal anastomoses between the mutants, were isolated from medium containing both mycostatin and cycloheximide at concentrations which did not permit growth of the mutants or the original parents. Both the mutants and the interspecies hybrids (ISH) derived from them expressed significant differences in pathogenicity to tomato and wheat from that expressed by the parents FOL and FG. Protein, esterase, pectinase and polyphenoloxidase electrophoretograms displayed unique patterns for the hybrids and the mutant parents. However, these patterns did not correlate with the pathogenicity expressions obtained. The study has demonstrated interspecies hybridizations between FOL(M) and FG(C) by hyphal anastomoses. Both the processes of mutation and hybridization altered the expression of pathogenicity in tomato and wheat The significance of these observations on the origin and development of new pathogenic forms is discussed.     

Isolation and characterization of a molecule stimulatory to growth of somatic embryos from early stage female gametophyte tissue of loblolly pine

Loblolly pine (LP, Pinus taeda) is the primary commercial species in southern forests of the US. Somatic embryogenesis (SE) is an effective technique to implement clonal tree production of high-value genotypes from breeding and genetic engineering programs. Unlike angiosperm embryos with attached cotyledons as seed storage organs, the diploid conifer embryo is surrounded by the unattached haploid female gametophyte (FG). The FG is not present in culture. This presents a dilemma if the FG produces necessary or regulatory compounds for embryo growth, since in culture these important compounds would be missing and would have to be added as supplements. We report here the direct evidence that extracts from early-stage FG indeed stimulate early-stage somatic embryo (SME) growth and multiplication, whereas extracts from late-stage FG inhibit early-stage SME growth. Furthermore, we have now isolated this stimulatory substance from early-stage FG tissue, and identified this substance as citric acid on the basis of NMR and mass spectrometry. We then demonstrated that topical application of citric acid to SMEs stimulates embryo colony growth at P = 0.05. Moreover, we find that there is a good correlation between the amount of citric acid isolated from FG tissue (65 nmoles per stage 2–3 FG) and the amount of citric acid that stimulates colony growth (25–50 nmoles) when applied topically to SMEs. This approach of isolating and characterizing a molecule from plant tissue, and investigating its role on SE processes can provide valuable information leading to further applications of these molecules to improve LP SE protocols.     

Facilitated aggregation of FG nucleoporins under molecular crowding conditions

Intrinsically disordered and phenylalanine–glycine‐rich nucleoporins (FG Nups) form a crowded and selective transport conduit inside the NPC that can only be transited with the help of nuclear transport receptors (NTRs). It has been shown in vitro that FG Nups can assemble into two distinct appearances, amyloids and hydrogels. If and how these phenomena are linked and if they have a physiological role still remains unclear. Using a variety of high‐resolution fluorescence and electron microscopic (EM) tools, we reveal that crowding conditions mimicking the NPC environment can accelerate the aggregation and amyloid formation speed of yeast and human FG Nups by orders of magnitude. Aggregation can be inhibited by NTRs, providing a rationale on how the cell might control amyloid formation of FG Nups. The superb spatial resolving power of EM also reveals that hydrogels are enlaced amyloid fibres, and these findings have implications for existing transport models and for NPC assembly.     

Stimulation of defense responses of in vitro potato plants by treatment with steroid glycosides under abiotic stresses

To elucidate the biological activity of furastanol glycosides (FG) as elicitors and their possible action mechanisms, antiradical properties of these compounds and their influence on in vitro potato (Solanum tuberosum L.) plants under abiotic stresses were studied. FG stimulated adaptive responses related to the activation of guaiacol-dependent peroxidase and suppression of lipid peroxidation (POL). Under normal conditions, in 30 min after plant treatment with 4.5 μM FG, the level of POL was decreased, activities of guaiacol- and ascorbate-dependent peroxidases increased, and activities of catalase and superoxide dismutase (SOD) remained unchanged. In 3 days after treatment with FG, POL was decreased by 20–60% in the leaves and by 50% in the roots. Under stress conditions (paraquat treatment for 11 days or hypothermia for 28 days), FG treatment reduced the level of POL by 20–40% as compared to the effects of stressors alone, which indicates FG elicitor properties. Antiradical properties of FG toward the hydroxyl radical were manifested in the reaction of deoxy-D-ribose destruction in water medium (without homogenate) within the concentration range from 4.5 to 65 μM; the effects toward the oxygen anion radical were insignificant. It is suggested that FG influence on generation of reactive oxygen species mediates the stimulation of potato plant defense responses to environmental factors.     

NaCl-induzierter Crassulaceensäurestoffwechsel bei einer weiteren Aizoacee: Carpobrotus edulis

Summary Carpobrotus edulis grown for 24 days in nutrient solution plus 400 mM of NaCl shows the typical CO₂ gas exchange reactions observed in CAM plants. Control plants grown in nutrient solution alone exhibit CO₂ gas exchange reactions typical for C₃ plants.

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Abkürzungen CAM Crassulaceensäurestoffwechsel - FG Frischgewicht     

Structure of Flower in Arabidopsis thaliana: Spatial Pattern Formation

Morphological analysis of flowers was carried out in Arabidopsis thaliana wild type plants and agamous and apetala2 mutants. No direct substitution of organs takes place in the mutants, since the number and position of organs in them do not correspond to the structure of wild type flower. In order to explain these data, a notion of spatial pattern formation in the meristem was introduced, which preceded the processes of appearance of organ primordia and formation of organs. Zones of acropetal and basipetal spatial pattern formation in the flower of wild type plants were postulated. It was shown that the acropetal spatial pattern formation alone took place in agamous mutants and basipetal spatial pattern formation alone, in apetala2 mutants. Different variants of flower structure are interpreted as a result of changes in the volume of meristem (space) and order of spatial pattern formation (time).     

Glycosylation defects activate filamentous growth Kss1 MAPK and inhibit osmoregulatory Hog1 MAPK

The yeast filamentous growth (FG) MAP kinase (MAPK) pathway is activated under poor nutritional conditions. We found that the FG‐specific Kss1 MAPK is activated by a combination of an O‐glycosylation defect caused by disruption of the gene encoding the protein O‐mannosyltransferase Pmt4, and an N‐glycosylation defect induced by tunicamycin. The O‐glycosylated membrane proteins Msb2 and Opy2 are both essential for activating the FG MAPK pathway, but only defective glycosylation of Msb2 activates the FG MAPK pathway. Although the osmoregulatory HOG (high osmolarity glycerol) MAPK pathway and the FG MAPK pathway share almost the entire upstream signalling machinery, osmostress activates only the HOG‐specific Hog1 MAPK. Conversely, we now show that glycosylation defects activate only Kss1, while activated Kss1 and the Ptp2 tyrosine phosphatase inhibit Hog1. In the absence of Kss1 or Ptp2, however, glycosylation defects activate Hog1. When Hog1 is activated by glycosylation defects in ptp2 mutant, Kss1 activation is suppressed by Hog1. Thus, the reciprocal inhibitory loop between Kss1 and Hog1 allows only one or the other of these MAPKs to be stably activated under various stress conditions.     

Population parameters for resistance to Fusarium graminearum and Fusarium verticillioides ear rot among large sets of early, mid-late and late maturing European maize (Zea mays L.) inbred lines

Infection of maize ears with Fusarium graminearum (FG) and Fusarium verticillioides (FV) reduces yield and quality by mycotoxin contamination. Breeding and growing varieties resistant to both Fusarium spp. is the best alternative to minimize problems. The objectives of our study were to draw conclusions on breeding for ear rot resistance by estimating variance components, heritabilities and correlations between resistances to FV and FG severity and to investigate different inoculation methods. In 2007 and 2008, three maturity groups (early, mid-late, late) each comprising about 150 inbred lines were tested in Germany, France, Italy, and Hungary according to their maturity group. They were silk channel inoculated by FG (early) and FV (all groups). In the late maturity group, additionally kernel inoculation was applied in a separate trial. The percentage of mycelium coverage on the ear was rated at harvest (0–100%). Significant (P < 0.01) genotypic variances of ear rot severity were found in all groups. Inoculation was superior to natural infection because of higher disease severities and heritabilities. In early maturing flints and dents, FG caused significantly (P < 0.01) higher ear rot severity than FV (61.7 and 55.1% FG vs. 18.2 and 11.1% FV ear rot severity, respectively). FV inoculation in Southern Europe (mid-late, late) resulted in similar means between 10.3 and 14.0%. Selection is complicated by significant (P < 0.01) genotype × environment interactions. Correlation between FG and FV severity was moderate in flints and dents (r = 0.59 and 0.49, respectively) but lines resistant to both fungi exist. We conclude that chances for selecting improved European elite maize material within the existing germplasms is promising by multi-environmental inoculation trials.     

Effect of Root Zone Temperature on the Mineral Composition of Xylem Sap and Plasma Membrane K+-Mg++-ATPase Activity of Grafted-Cucumber and -Figleaf Gourd Root Systems

Cucumber (Cucumis sativus L.) seedlings were grafted onto cucumber-(CG) or figleaf gourd- (FG, Cucurbita ficifolia Bouché)seedlings in order to determine the effect of solution temperature(12, 22, and 32°C) on the mineral composition of xylem sapand the plasma membrane K⁺-Mg⁺⁺-ATPase activities of the roots.Low solution temperature (12°C) lowered the concentrationof NO^–₃ and H₂PO^–₄ in xylem sap of CG plants butnot of FG plants. Concentrations of K⁺, Ca⁺⁺ and Mg⁺⁺ in xylemsap were less affected than anions by solution temperature.The plasma membrane of FG plants grown in 12°C solutiontemperature showed the highest K⁺- Mg⁺⁺-ATPase activity at allATP concentrations up to 3 mM and at low reaction temperatureup to 12°C, indicating resistance of figleaf gourd to lowroot temperature. (Received December 27, 1994; Accepted March 10, 1995)     

Histochemical Characterization and Distribution of Fiber Types in the Pectoralis Muscle of the Ostrich (Struthio camelus) and Emu (Dromaius novaehollandiae)

The muscle fibers of the pectoralis (M. pectoralis pars thoracicus) of a male and a female ostrich (Struthio camelus) and a male and a female emu (Dromaius novaehollandiae) were studied histochemically for succinate dehydrogenase and myofibrillar adenosine triphosphatase. Slow-tonic (ST), fast-twitch oxidative-glycolytic (FOG) and fast-twitch glycolytic (FG) fibers were approximately equal in number and distribution in the emu pectoralis examined. In the ostriches, both predominantly FG and approximately equal areas, were present. ST fibers were significantly (P ≤ 0.05) larger than the similarly (P ≥ 0.05) sized FG and FOG fibers in the female ostrich and emus. In the male ostrich ST fibers were smaller (P ≤ 0.05) than FG fibers, neither of which were significantly (P ≥ 0.05) different from FOG fibers. The ratites have the greatest percentage and widest distribution of ST fibers found in any avian pectoralis studied to date. This could represent the ancestoral avian pectoralis, neoteny or an effect of flightlessness. ST fibers are used in the maintenance of posture, which is probably the main role of the pectoralis in the emu. The predominantly FG areas of the ostrich are indicative of an additional function, namely, behavioural display. Sexual dimorphism in the ostrich pectoralis is strongly suggested.     

Histochemical Study of Xylem Cells in In Vitro Culture of Iris sibirica L.

In this study, lignin content data are presented for annual regenerant Iris sibirica plants, comparable to those in six-year-old intact plants. The structure of the shoots of Iris sibirica grown on artificial nutrient media was studied by the histochemical method. Features of the formation of the xylem in Iris sibirica on artificial nutrient media were revealed. Regenerants very quickly developed a complex system consisting of vascular bundles containing sieve tubes, vessels and tracheids, and hydrocyte systems. Hydrocytes of Iris sibirica were tracheids with lignified thickening, but, in contrast to tracheids and vessels of xylem (they are formed based on procambium or cambium—special lateral primary or secondary meristem), hydrocytes differentiated from the cells of permanent tissues (like phellogen), which probably possessed meristematic activity at the time of differentiation. In Iris sibirica hydrocytes covered the vascular bundle by the thick layer and strung along it up to a certain height. High lignin content in young regenerant Iris sibirica plants was due to the formation of the dense tissue from lignified tracheal elements. The study of the differentiation of xylem elements under controlled conditions can serve as a model for our understanding of wood formation processes.

     

CO2-Fixierungsreaktionen bei der Salzpflanze Mesembryanthemum crystallinum unter variierten Außenbedingungen

Summary The correlation of CO₂-fixation metabolism to various environmental conditions such as NaCl content of culture medium, air humidity and light intensity was investigated in the halophytic species Mesembryanthemum crystallinum. The data obtained demonstrate that a change in photosynthesis from C₃-pathway to crassulacean acid metabolism (CAM) is observed not only in NaCl treated plants as reported earlier but also in control plants grown in non-saline medium when environmental conditions (high light intensity, low air humidity) cause a water deficit in the leaves. It is suggested that water stress plays an important role in regulation between C₃- and CAM-pathway of photosynthesis in Mesembryanthemum crystallinum.

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Abkürzungen CAM Crassulaceensäurestoffwechsel - FG Frischgewicht - TG Trockengewicht - D Ende Dunkelphase - L Ende Lichtphase Herrn Prof. Dr. Otto Stocker zum 85. Geburtstag gewidmet     

Ecological application of biotic resistance to control the invasion of an invasive plant,Ageratina altissima

Biotic resistance is the ability of species in a community to limit the invasion of other species. However, biotic resistance is not widely used to control invasive plants. Experimental, functional, and modeling approaches were combined to investigate the processes of invasion by Ageratina altissima (white snakeroot), a model invasive species in South Korea. We hypothesized that (1) functional group identity would be a good predictor of biotic resistance to A. altissima, whereas a species identity effect would be redundant within a functional group, and (2) mixtures of species would be more resistant to invasion than monocultures. We classified 37 species of native plants into three functional groups based on seven functional traits. The classification of functional groups was based primarily on differences in life longevity and woodiness. A competition experiment was conducted based on an additive competition design with A. altissima and monocultures or mixtures of resident plants. As an indicator of biotic resistance, we calculated a relative competition index (RCI_avg) based on the average performance of A. altissima in a competition treatment compared with that of the control where only seeds of A. altissima were sown. To further explain the effect of diversity, we tested several diversity–interaction models. In monoculture treatments, RCI_avg of resident plants was significantly different among functional groups but not within each functional group. Fast‐growing annuals (FG1) had the highest RCI_avg, suggesting priority effects (niche pre‐emption). RCI_avg of resident plants was significantly greater in a mixture than in a monoculture. According to the diversity–interaction models, species interaction patterns in mixtures were best described by interactions between functional groups, which implied niche partitioning. Functional group identity and diversity of resident plant communities were good indicators of biotic resistance to invasion by introduced A. altissima, with the underlying mechanisms likely niche pre‐emption and niche partitioning. This method has most potential in assisted restoration contexts, where there is a desire to reintroduce natives or boost their population size due to some previous level of degradation.     

Nitrogen and carbohydrate storage in biennials originating from habitats of different resource availability

Four biennial species (Arctium tomentosum, Cirsium vulgare, Dipsacus sylvester and Daucus carota) which originate from habitats of different nutrient availability were investigated in a 2-year experiment in a twofactorial structured block design varying light (natural daylight versus shading) and fertilizer addition. The experiment was designed to study storage as reserve formation (competing with growth) or as accumulation (see Chapin et al. 1990). We show that (i) the previous definitions of storage excluded an important process, namely the formation of storage tissue. Depending on species, storage tissue and the filling process can be either a process of reserve formation, or a process of accumulation. (ii) In species representing low-resource habitats, the formation of a storage structure competes with other growth processes. Growth of storage tissue and filling with storage products is an accumulation process only in the high-resource plant Arctium tomentosum. We interpret the structural growth of low-resource plants in terms of the evolutionary history of these species, which have closely related woody species in the Mediterranean area. (iii) The use of storage products for early leaf growth determines the biomass development in the second season and the competitive ability of this species during growth with perennial species. (iv) The high-resource plant Arctium has higher biomass development under all conditions, i.e. plants of low-resource habitats are not superior under low-resource conditions. The main difference between high- and low-resource plants is that low-resource plants initiate flowering at a lower total plant internal pool size of available resources.     

蔗糖对列当科两种根部半寄生植物吸器发生的促进作用

吸器是寄生植物的特征器官,研究影响其发生的因素,有助于了解寄生关系的建立和调控过程。该研究以两种列当科(Orobanchaceae)根部半寄生植物甘肃马先蒿(Pedicularis kansuensis)和松蒿(Phtheirospermum japonicum)为材料,通过皿内培养试验,分析了蔗糖、DMBQ(2,6-二甲氧基-对-苯醌,一种高效的列当科根部半寄生植物吸器诱导化合物)和寄主植物诱导下两种根部半寄生植物吸器发生情况。结果表明：(1)蔗糖显著促进两种根部半寄生植物吸器发生,无寄主存在时,2%蔗糖处理使甘肃马先蒿和松蒿吸器发生率分别提高39.9%和20.2%。(2)蔗糖明显提升寄主植物对两种根部半寄生植物的吸器诱导水平,添加蔗糖后,寄主诱导的甘肃马先蒿单株吸器数和具木质桥的吸器比例分别增加5.7个/株和17.9%,松蒿吸器发生率和具木质桥的吸器比例分别提升76.7%和16.2%。(3)蔗糖对松蒿吸器发生的促进作用与已知吸器诱导化合物DMBQ相当,均能诱导50%以上的植株产生吸器。(4)培养基中添加4%蔗糖对两种根部半寄生植物的吸器诱导效果最好,其中甘肃马先蒿吸器发生率为56%...     

Dynamics of callose deposition and beta-1,3-glucanase expression during reproductive events in sexual and apomictic Hieracium

Callose accumulates in the walls of cells undergoing megasporogenesis during embryo sac formation in angiosperm ovules. Deficiencies in callose deposition have been observed in apomictic plants and causal linkages between altered callose deposition and apomictic initiation proposed. In apomictic Hieracium, embryo sacs initiate by sexual and apomictic processes within an ovule, but sexual development terminates in successful apomicts. Callose deposition and the events that lead to sexual termination were examined in different Hieracium apomicts that form initials pre- and post-meiosis. In apomictic plants, callose was not detected in initial cell walls and deficiencies in callose deposition were not observed in cells undergoing megasporogenesis. Multiple initial formation pre-meiosis resulted in physical distortion of cells undergoing megasporogenesis, persistence of callose and termination of the sexual pathway. In apomictic plants, callose persistence did not correlate with altered spatial or temporal expression of a β-1,3-glucanase gene (HpGluc) encoding a putative callose-degrading enzyme. Expression analysis indicated HpGluc might function during ovule growth and embryo sac expansion in addition to callose dissolution in sexual and apomictic plants. Initial formation pre-meiosis might therefore limit the access of HpGluc protein to callose substrate while the expansion of aposporous embryo sacs is promoted. Callose deposition and dissolution during megasporogenesis were unaffected when initials formed post-meiosis, indicating other events cause sexual termination. Apomixis in Hieracium is not caused by changes in callose distribution but by events that lead to initial cell formation. The timing of initial formation can in turn influence callose dissolution. Received: 18 April 2000 / Accepted: 10 July 2000     

Antagonistic Mechanism of Iturin A and Plipastatin A from Bacillus amyloliquefaciens S76-3 from Wheat Spikes against Fusarium graminearum

Controlling toxigenic Fusarium graminearum (FG) is challenging. A bacterial strain (S76-3, identified as Bacillus amyloliquefaciens) that was isolated from diseased wheat spikes in the field displayed strong antifungal activity against FG. Reverse-phase high performance liquid chromatography and electrospray ionization mass spectrometry analyses revealed that S76-3 produced three classes of cyclic lipopeptides including iturin, plipastatin and surfactin. Each class consisted of several different molecules. The iturin and plipastatin fractions strongly inhibited FG; the surfactin fractions did not. The most abundant compound that had antagonistic activity from the iturin fraction was iturin A (m/z 1043.35); the most abundant active compound from the plipastatin fraction was plipastatin A (m/z 1463.90). These compounds were analyzed with collision-induced dissociation mass spectrometry. The two purified compounds displayed strong fungicidal activity, completely killing conidial spores at the minimal inhibitory concentration range of 50 µg/ml (iturin A) and 100 µg/ml (plipastatin A). Optical and fluorescence microscopy analyses revealed severe morphological changes in conidia and substantial distortions in FG hyphae treated with iturin A or plipastatin A. Iturin A caused leakage and/or inactivation of FG cellular contents and plipastatin A caused vacuolation. Time-lapse imaging of dynamic antagonistic processes illustrated that iturin A caused distortion and conglobation along hyphae and inhibited branch formation and growth, while plipastatin A caused conglobation in young hyphae and branch tips. Transmission electron microscopy analyses demonstrated that the cell walls of conidia and hyphae of iturin A and plipastatin A treated FG had large gaps and that their plasma membranes were severely damaged and separated from cell walls.