Bernd Wollenweber

Water deficits and heat shock effects on photosynthesis of a transgenic Arabidopsis thaliana constitutively expressing ABP9, a bZIP transcription factor

The effects of water deficits (WD), heat shock (HS), and both (HSWD) on photosynthetic carbon- and light-use efficiencies together with leaf ABA content, pigment composition and expressions of stress- and light harvesting-responsive genes were investigated in ABP9 [ABA-responsive-element (ABRE) binding protein 9] transgenic Arabidopsis (5P2). WD, HS, and HSWD significantly decreased photosynthetic rate (A) and stomatal conductance (g(s)) in wild-type plants (WT). A and g(s) of 5P2 transgenic plants were slightly reduced by a single stress and were hardly modified by HSWD. Although A and electron transport rate (ETR) in 5P2 plants were depressed under optimal growth conditions (control) in relation to WT, they were enhanced under HS and HSWD. These results indicate that ABP9 transgenic plants are less susceptible to stress than the WT. In addition, the increased ABA contents in both WT and 5P2 plants in response to WD and/or HS stresses suggest that declines in A and g(s) might have been due to ABA-induced stomatal closure. Moreover, compared with WT, 5P2 plants exhibited higher ABA content, instantaneous water use efficiency (IWUE), Chl a/b, NPQ, and lower Chl/carotenoid ratios. Finally, altered expression of stress-regulated or light harvesting-responsive genes was observed. Collectively, our results indicate that constitutive expression of ABP9 improves the photosynthetic capacity of plants under stress by adjusting photosynthetic pigment composition, dissipating excess light energy, and elevating carbon-use efficiency as well as increasing ABA content, IWUE, and expression of stress-defensive genes, suggesting an important role of ABP9 in the regulation of plant photosynthesis under stress.

Implications of high-temperature events and water deficits on protein profiles in wheat (Triticum aestivum L. cv. Vinjett) grain.

Increased climatic variability is resulting in an increase of both the frequency and the magnitude of extreme climate events. Therefore, cereals may be exposed to more than one stress event in the growing season, which may ultimately affect crop yield and quality. Here, effects are reported of interaction of water deficits and/or a high‐temperature event (32°C) during vegetative growth (terminal spikelet) with either of these stress events applied during generative growth (anthesis) in wheat. Influence of combinations of stress on protein fractions (albumins, globulins, gliadins and glutenins) in grains and stress‐induced changes on the albumin and gliadin proteomes were investigated by 2‐DE and MS. The synthesis of individual protein fractions was shown to be affected by both the type and time of the applied stresses. Identified drought or high‐temperature‐responsive proteins included proteins involved in primary metabolism, storage and stress response such as late embryogenesis abundant proteins, peroxiredoxins and α‐amylase/trypsin inhibitors. Several proteins, e.g. heat shock protein and 14‐3‐3 protein changed in abundance only under multiple high temperatures.

Waterlogging pretreatment during vegetative growth improves tolerance to waterlogging after anthesis in wheat.

The interaction of multiple waterlogging events during vegetative growth (at the seven- and nine-leaf stage, and at heading) to a waterlogging event during the generative growth stage was studied in wheat (Triticum aestivum L. cv. Yangmai 9). Waterlogging before anthesis was found to effectively enhance tolerance to a waterlogging event after anthesis, as indicated by: (1) increasing net photosynthetsis (P(N)), stomatal conductance (g(s)) and transpiration (Tr) and maintaining high SPAD (soil plant analysis development) values; (2) enhancing use-efficiency of absorbed light energy in the stressed plants due to high maximum and actual quantum yield (F(v)/F(m), Φ(PSII)); (3) increasing activities of superoxide dismutase (SOD), ascorbate peroxidase (APX) and catalase (CAT); (4) enhancing dry matter accumulation after anthesis and its contribution to grain mass, which further resulting in significantly improved grain yields. The results indicate that hardening by waterlogging applied before anthesis can effectively improve the tolerance of wheat to waterlogging events occurring during the generative growth stage.

Multiple heat priming enhances thermo-tolerance to a later high temperature stress via improving subcellular antioxidant activities in wheat seedlings.

Seedlings of winter wheat (Triticum aestivum L.) were firstly twice heat-primed at 32/24 °C, and subsequently subjected to a more severe high temperature stress at 35/27 °C. The later high temperature stress significantly decreased plant biomass and leaf total soluble sugars concentration. However, plants experienced priming (PH) up-regulated the Rubisco activase B encoding gene RcaB, which was in accordance with the higher photosynthesis rate in relation to the non-primed plants (NH) under the later high temperature stress. In relation to NH, the major chlorophyll a/b-binding protein gene Cab was down-regulated in PH plants, implying a reduction of the light absorption to protect the photosystem II from excitation energy under high temperature stress. At the same time, under the later high temperature stress PH plants showed significantly higher actual photochemical efficiency, indicating an improvement of light use efficiency due to the priming pre-treatment. Under the later high temperature stress, PH could be maintained a better redox homeostasis than NH, as exemplified by the higher activities of superoxide dismutase (SOD) in chloroplasts and glutathione reductase (GR), and of peroxidase (POD) in mitochondria, which contributed to the lower superoxide radical production rate and malondialdehyde concentration in both chloroplasts and mitochondria. The improved antioxidant capacity in chloroplasts and mitochondria was related to the up-regulated expressions of Cu/Zn-SOD, Mn-SOD and GR in PH. Collectively, heat priming effectively improved thermo-tolerance of wheat seedlings subjected to a later high temperature stress, which could be largely ascribed to the enhanced anti-oxidation at the subcellular level.

Improved tolerance to drought stress after anthesis due to priming before anthesis in wheat (Triticum aestivum L.) var. Vinjett

Summary Physiological and proteome insights were determined into the alleviating effects of pre-anthesis drought priming on drought stress during post-anthesis, which will be of importance for future drought-tolerance studies in cereals.

Exploring abiotic stress on asynchronous protein metabolism in single kernels of wheat studied by NMR spectroscopy and chemometrics

Extreme climate events are being recognized as important factors in the effects on crop growth and yield. Increased climatic variability leads to more frequent extreme conditions which may result in crops being exposed to more than one extreme event within a growing season. The aim of this study was to examine the implications of different drought treatments on the protein fractions in grains of winter wheat using 1H nuclear magnetic resonance spectroscopy followed by chemometric analysis. Triticum aestivum L. cv. Vinjett was studied in a semi-field experiment and subjected to drought episodes either at terminal spikelet, during grain-filling or at both stages. Principal component trajectories of the total protein content and the protein fractions of flour as well as the 1H NMR spectra of single wheat kernels, wheat flour, and wheat methanol extracts were analysed to elucidate the metabolic development during grain-filling. The results from both the 1H NMR spectra of methanol extracts and the 1H HR-MAS NMR of single kernels showed that a single drought event during the generative stage had as strong an influence on protein metabolism as two consecutive events of drought. By contrast, a drought event at the vegetative growth stage had little effect on the parameters investigated. For the first time, 1H HR-MAS NMR spectra of grains taken during grain-filling were analysed by an advanced multiway model. In addition to the results from the chemical protein analysis and the 1H HR-MAS NMR spectra of single kernels indicating that protein metabolism is influenced by multiple drought events, the 1H NMR spectra of the methanol extracts of flour from mature grains revealed that the amount of fumaric acid is particularly sensitive to water deficits.

Bulk Carbohydrate Grain Filling of Barley β-Glucan Mutants Studied by 1H HR MAS NMR

ABSTRACT Temporal and genotypic differences in bulk carbohydrate accumulation in three barley genotypes differing in the content of mixed linkage β-(1→3),(1→4)-D-glucan (β-glucan) and starch were investigated using proton high-resolution, magic angle spinning, nuclear magnetic resonance (1H HR MAS NMR) during grain filling. For the first time, 1H HR MAS NMR spectra of flour from immature barley seeds are analyzed. Spectral assignments are made using two-dimensional (2D) NMR methods. Both α- and β-glucan biosynthesis were characterized by inspection of the spectra as well as by calibration to the reference methods for starch and β-glucan content. Starch was quantified with very good calibrations to the α-(1→4) peak (5.29–5.40 ppm) and the region 3.67–3.83 ppm covering starch glycopyranosidic protons from H5 and H6. In contrast, the spectral inspection of the β-anomeric region 4.45–4.85 ppm showed unexpected lack of intensity in the high β-glucan mutant lys5f at seed maturity, resulting in poor calibration ...

Changes in carbon and nitrogen allocation, growth and grain yield induced by arbuscular mycorrhizal fungi in wheat (Triticum aestivum L.) subjected to a period of water deficit

Drought is a major abiotic factor limiting agricultural crop production. One of the effective ways to increase drought resistance in plants could be to optimize the exploitation of symbiosis with arbuscular mycorrhizal fungi (AMF). Hypothesizing that alleviation of water deficits by AMF in wheat will help maintain photosynthetic carbon-use, we studied the role of AMF on gas-exchange, light-use efficiencies, carbon/nitrogen ratios and growth and yield parameters in the contrasting wheat (Triticum aestivum L.) cultivars ‘Vinjett’ and ‘1110’ grown with/without AMF symbiosis. Water deficits applied at the floret initiation stage significantly decreased rates of photosynthetic carbon gain, transpiration and stomatal conductance in the two wheat cultivars. AMF increased the rates of photosynthesis, transpiration and stomatal conductance under drought conditions. Water deficits decreased electron transport rate and increased non-photochemical quenching (NPQ) in ‘1110’ but not in ‘Vinjett’. With AMF, nitrogen concentrations increased in roots of both cultivars, but decreased in grains of ‘Vinjett’ and in side-tiller grains of ‘1110’ regardless of water status. With water deficits, AMF colonization increased plant height in both cultivars. AMF also increased biomass and grain yield in ‘1110’ but not in ‘Vinjett’. The results showed that the improvements in growth and yield were the results of AMF-mediated increases in photosynthesis during drought stress and that the alleviating effect of AMF depended on the wheat cultivar.

Are ant feces nutrients for plants? A metabolomics approach to elucidate the nutritional effects on plants hosting weaver ants

Weaver ants (genus Oecophylla) are tropical carnivorous ant species living in high numbers in the canopies of trees. The ants excrete copious amounts of fecal matter on leaf surfaces, and these feces may provide nutrients to host trees. This hypothesis is supported by studies of ant-plant interactions involving other ant species that have demonstrated the transfer of nutrients from ants to plants. In this 7-months study, a GC–MS-based metabolomics approach along with an analysis of total nitrogen and carbon levels was used to study metabolic changes in ant-hosting Coffea arabica plants compared with control plants. The results showed elevated levels of total nitrogen, amino acids, fatty acids, caffeine, and secondary metabolites of the phenylpropanoid pathway in leaves from ant-hosting plants. Minor effects were observed for sugars, whereas little or no effect was observed for organic acids, despite the fact that lower levels of total carbon were found in ant-hosting plants. The increased levels of total nitrogen, amino acids, fatty acids and caffeine and the decreased total carbon were consistent with changes observed in plants grown with an increased supply of nitrogen-containing nutrients. The up-regulation of the phenylpropanoid pathway could indicate biotic stress and/or nutrient deficiency. However, because nutrient deficiency was contradicted by the remaining results, this pathway up-regulation was ascribed to biotic stress caused by the physical presence of the weaver ants.

Evolutionary Pattern of N-Glycosylation Sequon Numbers in Eukaryotic ABC Protein Superfamilies

Many proteins contain a large number of NXS/T sequences (where X is any amino acid except proline) which are the potential sites of asparagine (N) linked glycosylation. However, the patterns of occurrence of these N-glycosylation sequons in related proteins or groups of proteins and their underlying causes have largely been unexplored. We computed the actual and probabilistic occurrence of NXS/T sequons in ABC protein superfamilies from eight diverse eukaryotic organisms. The ABC proteins contained significantly higher NXS/T sequon numbers compared to respective genome-wide average, but the sequon density was significantly lower owing to the increase in protein size and decrease in sequon specific amino acids. However, mammalian ABC proteins have significantly higher sequon density, and both serine and threonine containing sequons (NXS and NXT) have been positively selected—against the recent findings of only threonine specific Darwinian selection of sequons in proteins. The occurrence of sequons was positively correlated with the frequency of sequon specific amino acids and negatively correlated with proline and the NPS/T sequences. Further, the NPS/T sequences were significantly higher than expected in plant ABC proteins which have the lowest number of NXS/T sequons. Accordingly, compared to overall proteins, N-glycosylation sequons in ABC protein superfamilies have a distinct pattern of occurrence, and the results are discussed in an evolutionary perspective.