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Publications - Cell and Metabolic Biology

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Publications

Frolov, A.; Bilova, T.; Paudel, G.; Berger, R.; Balcke, G. U.; Birkemeyer, C.; Wessjohann, L. A.; Early responses of mature Arabidopsis thaliana plants to reduced water potential in the agar-based polyethylene glycol infusion drought model J. Plant Physiol. 208, 70-83, (2017) DOI: 10.1016/j.jplph.2016.09.013

Drought is one of the most important environmental stressors resulting in increasing losses of crop plant productivity all over the world. Therefore, development of new approaches to increase the stress tolerance of crop plants is strongly desired. This requires precise and adequate modeling of drought stress. As this type of stress manifests itself as a steady decrease in the substrate water potential (ψw), agar plates infused with polyethylene glycol (PEG) are the perfect experimental tool: they are easy in preparation and provide a constantly reduced ψw, which is not possible in soil models. However, currently, this model is applicable only to seedlings and cannot be used for evaluation of stress responses in mature plants, which are obviously the most appropriate objects for drought tolerance research. To overcome this limitation, here we introduce a PEG-based agar infusion model suitable for 6–8-week-old A. thaliana plants, and characterize, to the best of our knowledge for the first time, the early drought stress responses of adult plants grown on PEG-infused agar. We describe essential alterations in the primary metabolome (sugars and related compounds, amino acids and polyamines) accompanied by qualitative and quantitative changes in protein patterns: up to 87 unique stress-related proteins were annotated under drought stress conditions, whereas further 84 proteins showed a change in abundance. The obtained proteome patterns differed slightly from those reported for seedlings and soil-based models.
Publications

Hazman, M.; Hause, B.; Eiche, E.; Riemann, M.; Nick, P.; Different forms of osmotic stress evoke qualitatively different responses in rice J. Plant Physiol. 202, 45-56, (2016) DOI: 10.1016/j.jplph.2016.05.027

Drought, salinity and alkalinity are distinct forms of osmotic stress with serious impacts on rice productivity. We investigated, for a salt-sensitive rice cultivar, the response to osmotically equivalent doses of these stresses. Drought, experimentally mimicked by mannitol (single factor: osmotic stress), salinity (two factors: osmotic stress and ion toxicity), and alkalinity (three factors: osmotic stress, ion toxicity, and depletion of nutrients and protons) produced different profiles of adaptive and damage responses, both locally (in the root) as well as systemically (in the shoot). The combination of several stress factors was not necessarily additive, and we even observed cases of mitigation, when two (salinity), or three stressors (alkalinity) were compared to the single stressor (drought). The response to combinations of individual stress factors is therefore not a mere addition of the partial stress responses, but rather represents a new quality of response. We interpret this finding in a model, where the output to signaling molecules is not determined by their abundance per se, but qualitatively depends on their adequate integration into an adaptive signaling network. This output generates a systemic signal that will determine the quality of the shoot response to local concentrations of ions.
Publications

Klopotek, Y.; Haensch, K.-T.; Hause, B.; Hajirezaei, M.-R.; Druege, U.; Dark exposure of petunia cuttings strongly improves adventitious root formation and enhances carbohydrate availability during rooting in the light J. Plant Physiol. 167, 547-554, (2010) DOI: 10.1016/j.jplph.2009.11.008

The effect of temporary dark exposure on adventitious root formation (ARF) in Petunia×hybrida ‘Mitchell’ cuttings was investigated. Histological and metabolic changes in the cuttings during the dark treatment and subsequent rooting in the light were recorded. Excised cuttings were exposed to the dark for seven days at 10°C followed by a nine-day rooting period in perlite or were rooted immediately for 16 days in a climate chamber at 22/20°C (day/night) and a photosynthetic photon flux density (PPFD) of 100 μmol m−2 s−1. Dark exposure prior to rooting increased, accelerated and synchronized ARF. The rooting period was reduced from 16 days (non-treated cuttings) to 9 days (treated cuttings). Under optimum conditions, despite the reduced rooting period, dark-exposed cuttings produced a higher number and length of roots than non-treated cuttings. An increase in temperature to 20 °C during the dark treatment or extending the cold dark exposure to 14 days caused a similar enhancement of root development compared to non-treated cuttings. Root meristem formation had already started during the dark treatment and was enhanced during the subsequent rooting period. Levels of soluble sugars (glucose, fructose and sucrose) and starch in leaf and basal stem tissues significantly decreased during the seven days of dark exposure. This depletion was, however, compensated during rooting after 6 and 24h for soluble sugars in leaves and the basal stem, respectively, whereas the sucrose level in the basal stem was already increased at 6 h. The association of higher carbohydrate levels with improved rooting in previously dark-exposed versus non-treated cuttings indicates that increased post-darkness carbohydrate availability and allocation towards the stem base contribute to ARF under the influence of dark treatment and provide energy for cell growth subject to a rising sink intensity in the base of the cutting.
Publications

Cenzano, A.; Abdala, G.; Hause, B.; Cytochemical immuno-localization of allene oxide cyclase, a jasmonic acid biosynthetic enzyme, in developing potato stolons J. Plant Physiol. 164, 1449-1456, (2007) DOI: 10.1016/j.jplph.2006.10.007

The involvement of jasmonates in the tuber development has been proved by the presence of many of these compounds in potato stolons, modification of their levels during the transition of the stolon into tuber, and induction of cell expansion upon exogenous jasmonates treatment. However, to date there is only little evidence of the presence of the jasmonic acid-biosynthetic enzymes in stolons or young tubers. As allene oxide cyclase represents the major control point for jasmonic acid biosynthesis, we studied the occurrence of allene oxide cyclase by immunological approaches in the early stages of tuber formation. In developing stolons, allene oxide cyclase as well as lipoxygenase were clearly detectable, but their levels did not change during development. Jasmonic acid treatment for 24 h, however, increased lipoxygenase and allene oxide cyclase protein levels in both developmental stages analyzed. In longitudinal sections of stolons of stages 1 and 2, allene oxide cyclase and lipoxygenase occurred in the apex and along the stolon axis. Allene oxide cyclase was clearly detectable in epidermal, cortical and pith parenchymatic cells, showing the highest levels in vascular tissues surrounding cells. Lipoxygenase was mainly located in the parenchymatic cortex cells. The occurrence of allene oxide cyclase in stolons together with the previous identification of jasmonates from developing stolons reveals that these organs are capable to synthesize and metabolize jasmonates.
Publications

Wasternack, C.; Stenzel, I.; Hause, B.; Hause, G.; Kutter, C.; Maucher, H.; Neumerkel, J.; Feussner, I.; Miersch, O.; The wound response in tomato – Role of jasmonic acid J. Plant Physiol. 163, 297-306, (2006) DOI: 10.1016/j.jplph.2005.10.014

Plants respond to mechanical wounding or herbivore attack with a complex scenario of sequential, antagonistic or synergistic action of different signals leading to defense gene expression. Tomato plants were used as a model system since the peptide systemin and the lipid-derived jasmonic acid (JA) were recognized as essential signals in wound-induced gene expression. In this review recent data are discussed with emphasis on wound-signaling in tomato. The following aspects are covered: (i) systemin signaling, (ii) JA biosynthesis and action, (iii) orchestration of various signals such as JA, H2O2, NO, and salicylate, (iv) local and systemic response, and (v) amplification in wound signaling. The common occurrence of JA biosynthesis and systemin generation in the vascular bundles suggest JA as the systemic signal. Grafting experiments with JA-deficient, JA-insensitive and systemin-insensitive mutants strongly support this assumption.
Publications

Camacho-Cristóbal, J. J.; Lunar, L.; Lafont, F.; Baumert, A.; González-Fontes, A.; Boron deficiency causes accumulation of chlorogenic acid and caffeoyl polyamine conjugates in tobacco leaves J. Plant Physiol. 161, 879-881, (2004) DOI: 10.1016/j.jplph.2003.12.003

The effects of boron (B) deficiency on carbohydrate concentrations and the pattern of phenolic compounds were studied in leaves of tobacco plants (Nicotiana tabacum L.). Plants grown under B deficiency showed a notable increase in leaf carbohydrates and total phenolic compounds when compared to controls. The qualitative composition of phenolics was analyzed by HPLC-mass spectrometry. The level of caffeate conjugates (i.e., chlorogenic acid) increased in B-deficient plants. In addition, the accumulation of two caffeic acid amides (N-caffeoylputrescine and putative dicaffeoylspermidine) was observed.
Publications

Isayenkov, S.; Fester, T.; Hause, B.; Rapid determination of fungal colonization and arbuscule formation in roots of Medicago truncatula using real-time (RT) PCR J. Plant Physiol. 161, 1379-1383, (2004) DOI: 10.1016/j.jplph.2004.04.012

The quantifications of root colonization and symbiotic activity in the arbuscular mycorrhizal (AM) association of Medicago truncatula and Glomus intraradices were performed by quantitative polymerase chain reaction (real-time PCR). A strong correlation between fungal colonization of the root system and the amounts of fungal rDNA and rRNA were shown. In contrast, the transcript levels of the AM-specific phosphate transporter 4 from M. truncatula (MtPT4) correlate with arbuscule formation rather than with fungal colonization. These results suggest (i) that real-time PCR assay is a rapid, useful, and accurate method for the determination of arbuscular mycorrhizal features, (ii) that the amount of fungal rDNA or rRNA is a good parameter to estimate fungal colonization, and (iii) that it is necessary to evaluate the amount of other transcripts—like the MtPT4 transcript—to obtain additional information about the symbiotic state of the colonized root system.
Publications

Vierheilig, H.; Maier, W.; Wyss, U.; Samson, J.; Strack, D.; Piché, Y.; Cyclohexenone derivative- and phosphate-levels in split-root systems and their role in the systemic suppression of mycorrhization in precolonized barley plants J. Plant Physiol. 157, 593-599, (2000) DOI: 10.1016/S0176-1617(00)80001-2

In a split-root system root colonization by the arbuscular mycorrhizal fungus Glomus mosseae on one side is reduced when roots on the other side are already colonized by G. mosseae. Root colonization by arbuscular mycorrhizal fungi enhances the P-status of plants, thus the observed suppressional effect on further root colonization in precolonized barley plants could be P-level regulated. Split-root systems allow to separate plant mediated P-effects on root colonization by arbuscular mycorrhizal fungi from direct P-effects on arbuscular mycorrhizal fungi. By adding a KH2PO4-solution to one side of the split-root system of non-mycorrhizal control plants, higher P-levels were obtained as in split-root systems of G. mosseae precolonized plants. Subsequent inoculation with G. mosseae of the P-supplied and the precolonized plants resulted in an inhibition of root colonization in the precolonized plants, but not in the P-supplied plants, discarding the enhanced P-level as the responsible factor for the observed suppression. Cyclohexenone derivatives are secondary plant compounds only found in roots of mycorrhizal plants. Analysis of cyclohexenone derivatives in mycorrhizal and non-mycorrhizal roots in split-root systems revealed that cyclohexenone derivatives can be detected in mycorrhizal roots, but not in non-mycorrhizal roots of mycorrhizal plants. The presented results show clearly that cyclohexenone derivatives are not systemically accumulated and that the P-levels are not the responsible factors for the observed systemic suppression of mycorrhization in roots of precolonized barley plants.
Publications

Hause, B.; Vörös, K.; Kogel, K.-H.; Besser, K.; Wasternack, C.; A Jasmonate-responsive Lipoxygenase of Barley Leaves is Induced by Plant Activators but not by Pathogens J. Plant Physiol. 154, 459-462, (1999) DOI: 10.1016/S0176-1617(99)80283-1

Using the recently isolated eDNA clone LOX2 : Hv : 1 which codes for the most abundant jasmonateinducible lipoxygenase (LOX) in barley leaves (Vörös et al., 1998), we analysed the capability of different activators of systemic activated resistance (SAR) to induce the expression of that LOX. Upon treatment of barley leaves with salicylate, 2,6-dichloroisonicotinic acid and benzo-(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester, all these compounds were able to induce the expression of the LOX2 : Hv : 1 gene, whereas upon infection with the powdery mildew fungus (Blumeria graminis f. sp. hordei) mRNA accumulation was not detectable in compatible or in incompatible interactions. The induction of the LOX2 : Hv : 1 protein by SAR activators and the expression of different sets of genes induced by jasmonate and salicylate, respectively, are discussed in relation to defense responses against pathogenic fungi.
Publications

Wasternack, C.; Ortel, B.; Miersch, O.; Kramell, R.; Beale, M.; Greulich, F.; Feussner, I.; Hause, B.; Krumm, T.; Boland, W.; Parthier, B.; Diversity in octadecanoid-induced gene expression of tomato J. Plant Physiol. 152, 345-352, (1998) DOI: 10.1016/S0176-1617(98)80149-1

In tomato plants wounding leads to up-regulation of various plant defense genes via jasmonates (Ryan, 1992; Bergey et al., 1996). Using this model system of jasmonic acid (JA) signalling, we analyzed activity of octadecanoids to express JA-responsive genes. Leaf treatments were performed with naturally occurring octadecanoids and their molecular mimics such as coronatine or indanone conjugates. JA responses were recorded in terms of up- or down-regulation of various genes by analyzing transcript accumulation, and at least partially in vitro translation products and polypeptide pattern of leaf extracts. The data suggest: (i) 12-Oxo-phytodienoic acid and other intermediates of the octadecanoid pathway has to be ß-oxidized to give a JA response, (ii) Octadecanoids which can not be ß-oxidized are inactive, (iii) JA, its methyl ester (JM), and its amino acid conjugates are most active signals in tomato leaves leading to up regulation of mainly wound-inducible genes and down-regulation of mainly <house-keeping> genes, (iv) Some compounds carrying a JA/JM- or JA amino acid conjugate-like structure induce/repress only a subset of genes suggesting diversity of JA signalling.
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