Stress Response Monitoring of Photoautotrophic Higher Plant Suspension Cultures by Fluorescence Imaging for High-Throughput Toxic Compound Screening

Photoautotrophic suspension cultures have been established from various model and crop plants and proved to be valuable and robust experimental system to assess coordinated responses of primary and secondary metabolism to metabolic and stress related signals. The use of suspension cultures combines the ease of handling microalgae in microtiter plates with the advantage of testing physiological responses of higher plants, notably in combination with the assessment of the response of photosynthetic activity by PAM chlorophyll fluorescence imaging as well as monitoring changes in secondary metabolite production and ROS formation by steady state fluorescence of plant fluorophores or introduced fluorescent probes. Photoautotrophic cultures provide various advantages as fast, highly sensitive, robust and high-through-put experimental system for screening and characterization of the impact of toxic compounds on higher plants. This opinion article discusses and critically evaluates the potential of photoautotrophic cultures of higher plants in combination with fluorescence imaging assays in microtiter plates as a complement to existing guidelines for testing the toxicity of chemicals in plants

Metabolic control of tobacco pollination by sugars and invertases

Pollination in flowering plants is initiated by germination of pollen grains on stigmas followed by fast growth of pollen tubes representing highly energy-consuming processes. The symplastic isolation of pollen grains and tubes requires import of Suc available in the apoplast. We show that the functional coupling of Suc cleavage by invertases and uptake of the released hexoses by monosaccharide transporters are critical for pollination in tobacco (Nicotiana tabacum). Transcript profiling, in situ hybridization, and immunolocalization of extracellular invertases and two monosaccharide transporters in vitro and in vivo support the functional coupling in supplying carbohydrates for pollen germination and tube growth evidenced by spatiotemporally coordinated expression. Detection of vacuolar invertases in maternal tissues by these approaches revealed metabolic cross talk between male and female tissues and supported the requirement for carbohydrate supply in transmitting tissue during pollination. Tissue-specific expression of an invertase inhibitor and addition of the chemical invertase inhibitor miglitol strongly reduced extracellular invertase activity and impaired pollen germination. Measurements of (competitive) uptake of labeled sugars identified two import pathways for exogenously available Suc into the germinating pollen operating in parallel: direct Suc uptake and via the hexoses after cleavage by extracellular invertase. Reduction of extracellular invertase activity in pollen decreases Suc uptake and severely compromises pollen germination. We further demonstrate that Glc as sole carbon source is sufficient for pollen germination, whereas Suc is supporting tube growth, revealing an important regulatory role of both the invertase substrate and products contributing to a potential metabolic and signaling-based multilayer regulation of pollination by carbohydrates.Peer Reviewe

Fast track to nanomaterials: microwave assisted synthesis in ionic liquid media

Herein we present a general approach to metal and metal oxide nanoparticles using simple metal salts as starting materials. The reducing agent can be delivered in the form of the anion incorporated into the metal precursor respectively ionic liquid. Exemplary we demonstrate the synthesis of Cu and Ag as well as ZnO and NiO nanoparticles generated either from acetate or carbonate salts. All particles are synthesised by microwave heating without the necessity of inert conditions. Two different types of ionic liquids have been used as reaction media - tetra-n-butylphosphonium acetate (n-Bu4POAc) and 1-butyl-2,3-dimethylimidazolium N,N-bis(trifluoromethylsulfonyl)imid (bmmim NTf2). In this case, the choice of the ionic liquid seems to have significant influence on the size, shape and dispersity of the synthesised particles. It is clearly shown that the acetate anion present in all reaction mixtures can act as an inexpensive and nontoxic reducing agent. The final products in solid, liquid and gaseous phase have been characterised by XRD, TEM, NMR, FT-IR and online gas-phase MS

Anther-specific carbohydrate supply and restoration of metabolically engineered male sterility

Male-sterile plants are used in hybrid breeding as well as for gene confinement for genetically modified plants in field trials and agricultural production. Apart from naturally occurring mutations leading to male sterility, biotechnology has added new possibilities for obtaining male-sterile plants, although so far only one system is used in practical breeding due to limitations in propagating male-sterile plants without segregations in the next generation or insufficient restoration of fertility when fruits or seeds are to be harvested from the hybrid varieties. Here a novel mechanism of restoration for male sterility is presented that has been achieved by interference with extracellular invertase activity, which is normally specifically expressed in the anthers to supply the developing microspores with carbohydrates. Microspores are symplastically isolated in the locular space of the anthers, and thus an unloading pathway of assimilates via the apoplasmic space is mandatory for proper development of pollen. Antisense repression of the anther-specific cell wall invertase or interference with invertase activity by expressing a proteinacious inhibitor under the control of the anther-specific invertase promoter results in a block during early stages of pollen development, thus causing male sterility without having any pleiotropic effects. Restoration of fertility was successfully achieved by substituting the down-regulated endogenous plant invertase activity by a yeast invertase fused to the N-terminal portion of potato-derived vacuolar protein proteinase II (PiII–ScSuc2), under control of the orthologous anther-specific invertase promoter Nin88 from tobacco. The chimeric fusion PiII–ScSuc2 is known to be N-glycosylated and efficiently secreted from plant cells, leading to its apoplastic location. Furthermore, the Nin88::PiII-ScSuc2 fusion does not show effects on pollen development in the wild-type background. Thus, such plants can be used as paternal parents of a hybrid variety, thereby the introgression of Nin88::PiII-ScSuc2 to the hybrid is obtained and fertility is restored. In order to broaden the applicability of this male sterility/restoration system to other plant species, a phylogenic analysis of plant invertases(β-fructofuranosidases) and related genes of different species was carried out. This reveals a specific clustering of the cell wall invertases with anther-specific expression for dicotyl species and another cluster for monocotyl plants. Thus, in both groups of plants, there seems to be a kind of co-evolution, but no recent common ancestor of these members of the gene family. These findings provide a helpful orientation to classify corresponding candidate genes in further plant species, in addition to the species analysed so far (Arabidopsis, tobacco, tomato, potato, carrots, rice, and wheat)

A transnational and holistic breeding approach is needed for sustainable wheat production in the Baltic Sea region

The Baltic Sea is one of the largest brackish water bodies in the world. Eutrophication is a major concern in the Baltic Sea due to the leakage of nutrients to the sea with agriculture being the primary source. Wheat (Triticum aestivum L.) is the most widely grown crop in the countries surrounding the Baltic Sea and thus promoting sustainable agriculture practices for wheat cultivation will have a major impact on reducing pollution in the Baltic Sea. This approach requires identifying and addressing key challenges for sustainable wheat production in the region. Implementing new technologies for climate-friendly breeding and digital farming across all surrounding countries should promote sustainable intensification of agriculture in the region. In this review, we highlight major challenges for wheat cultivation in the Baltic Sea region and discuss various solutions integrating transnational collaboration for pre-breeding and technology sharing to accelerate development of low input wheat cultivars with improved host plant resistance to pathogen and enhanced adaptability to the changing climate.Peer reviewe

A Novel High-Affinity Sucrose Transporter Is Required for Virulence of the Plant Pathogen Ustilago maydis

A novel, high-affinity sucrose transporter identified in the plasma membrane of the plant pathogen Ustilago maydis is essential for fungal virulence and successful infection of maize