Ralf Kaldenhoff

• Prof. Dr.
• Professor (Full) at Technical University of Darmstadt

Photosynthetic activity can be monitored using pulse amplitude modulated (PAM) fluorescence or gas exchange. While PAM provides insight into the light-dependent reactions, gas exchange reflects CO2 fixation and water balance. Accurate, non-invasive prediction of photosynthetic performance under varying conditions is highly relevant for phenotyping...

This study investigates the effects of ultra-high light intensities and varying light spectra on the photosynthetic efficiency and growth of maize (Zea mays saccharata). Photosynthetic rates, transpiration, stomatal conductance, and leaf temperature were measured under white light, monochromatic light, and their combinations. Assimilation rates inc...

Microalgae, small single or multicellular photosynthetic active organisms, could be a component to solve our urgent global challenges. This review provides a concise introduction to biology and applications of microalgae. On an example from our own scientific studies, we illustrate how these organisms could replace respectively optimize carbon prod...

Background Information Aquaporins are H 2 O‐permeable membrane protein pores. However, some aquaporins are also permeable to other substances such as CO 2 . In higher plants, overexpression of such aquaporins has already led to an enhanced photosynthetic performance due to improved CO 2 mesophyll conductance. In this work, we investigated the effec...

Photosynthetic Cyanobacteria can be used as a chassis for different synthetic biology approaches. However, quantitative comparison of tools for engineering, such as those for heterologous gene expression, is often not available. Here, we report the comparative quantification of heterologous protein production in Synechococcussp.PCC 7002 regarding p...

The increased use of wireless technology causes a significant exposure increase for all living organisms to radio frequency electromagnetic fields (RF-EMF). This comprises bacteria, animals, and also plants. Unfortunately, our understanding of how RF-EMF influences plants and plant physiology remains inadequate. In this study, we examined the effec...

Modern agriculture demands for comprehensive information about the plants themselves. Conventional chemistry-based analytical methods—due to their low throughput and high associated costs—are no longer capable of providing these data. In recent years, remote reflectance-based characterisation has become one of the most promising solutions for rapid...

Modern agriculture demands for comprehensive information about the plant itself. Conventional chemistry-based analytical methods-due to their low throughput and high associated cost-are no longer capable of providing these data. In recent years, remote reflectance-based characterization has developed as one of the most promising solutions for rapid...

To meet the increasing demand for mass-produced microalgae, production processes must be optimised and monitored. The herein described optical sensors provide an instantaneous and direct opportunity to monitor biological and biochemical information for microalgae cultivation. Combination of expertise in natural science and engineering, as well as t...

The unicellular green alga Chlamydomonas reinhardtii has been used as a model organism in different research fields for the last 70 years. A variety of methods have been developed for its genetic modification, ranging from classical mutagenesis to the introduction, integration, and expression of foreign genes as well as cutting-edge genome editing...

In Chlamydomonas reinhardtii, ketocarotenoid biosynthesis is limited to the diploid zygospore stage. In this study, we attempted to engineer the ketocarotenoid pathway into Chlamydomonas haploid vegetative green cells by overexpressing the key enzyme ß-carotene ketolase (CrBKT). We chose strain CC-4102 for the approach; competitive pathways, α-caro...

Aquaporins are important and well-studied water channel membrane proteins. However, being membrane proteins, sample preparation for functional analysis is tedious and time-consuming. In this paper, we report a new approach for the co-translational insertion of two aquaporins from Escherichia coli and Nicotiana tabacum using the CFPS system. This wa...

Aquaporins are key proteins in regulating water transport, plant growth and development. In this study, we investigated the function of plasma membrane intrinsic proteins (PIPs) in both yeast (Saccharomyces cerevisiae) and rice (Oryza sativa cv. Nipponbare). Three OsPIP1s (OsPIP1;1, OsPIP1;2 and OsPIP1;3) and four OsPIP2s (OsPIP2;1, OsPIP2;3, OsPIP...

The present invention relates to a method for the control of mistletoe, and the use of folate biosynthesis inhibiting drug for combating mistletoe, in particular the dwarf mistletoe (Arceuthobium spp.).

Aquaporins in plants show more abundant and greater diversity than aquaporins in bacteria and animals. This unique characteristic provided versatile tool boxes for plants, dealing with environmental changes, which overcome the disadvantage of immobility. Aquaporins were first known for their function as water channel proteins. Later on, more and mo...

Plasma membrane intrinsic proteins (PIPs) are one subfamily of aquaporins that mediate the transmembrane transport of water. To reveal their function in poplar, we generated transgenic poplar plants in which the translation of PIP genes was downregulated by RNA interference investigated these plants with a comprehensive leaf plasma membrane proteom...

Atmospheric CO2 impacts all aspects of plant development. It has changed in the past and is predicted to change further on. Studies on the response of crop plants to low and elevated CO2 concerning growth, productivity and physiological processes are intense. In contrast, the molecular mechanisms of cellular CO2 exchange are still under discussion....

Affymetrix data based on GeneChip® AraGene-1.0 ST arrays (Affymetrix, USA).

Differentially expressed genes in atpip1;2-1 mutant line based on the t-test and SAM.

R flow for differential gene expression analysis.

Results from CO2 diffusion studies and characterization of Arabidopsis thaliana aquaporin AtPIP1;2 T-DNA insertion lines support the idea that specific aquaporins facilitate the diffusion of CO2 through biological membranes. However, their function as CO2 diffusion facilitators in plant physiology is still a matter of debate. Assuming that a lack o...

Black lipid bilayers, a general model system for biomembranes were studied for diffusion rates of small molecules such as water or CO2 using advanced analysis techniques and cell free synthesized proteins. We provide evidence that by simple insertion of proteins or sterols the diffusion rates of water or those of CO2 decrease. Insertion of cell fre...

Stomata, the microscopic pores on the surface of the aerial parts of plants, are bordered by two specialized cells, known as guard cells, which control the stomatal aperture according to endogenous and environmental signals. Like most movements occurring in plants, the opening and closing of stomata are based on hydraulic forces. During opening, th...

An approach for designing individual expression environments that reduce or prevent protein aggregation and precipitation is described. Inefficient folding of difficult proteins in unfavorable translation environments can cause significant losses of overexpressed proteins as precipitates or inclusion bodies. A number of chemical chaperones includin...

We functionally characterized the grape (Vitis vinifera) VvPIP2;4N (for Plasma membrane Intrinsic Protein) aquaporin gene. Expression of VvPIP2;4N in Xenopus laevis oocytes increased their swelling rate 54-fold. Northern blot and quantitative reverse transcription-polymerase chain reaction analyses showed that VvPIP2;4N is the most expressed PIP2 g...

We demonstrate that membranes consisting of certain triblock-copolymers were tight for CO₂. Using a novel approach, we provide evidence for aquaporin facilitated CO₂ diffusion. Plant aquaporins obtained from heterologous expression were inserted into triblock copolymer membranes. These were employed to separate a chamber with a solution maintaining...

The parasitic plant species Cuscuta reflexa and Phelipanche aegyptiaca have independently developed parasitism, the former parasitizing on shoots and the latter attaching to roots. Regardless of these differences, the two species use similar organs, termed haustoria, to attach to the host plant. In this study, we show that this morphological simila...

It is a general opinion that the Meyer-Overton correlation determines bio-membrane transport of hydrophobic molecules. This so called “Overton rule” says that the easier it is for a chemical to dissolve in a lipid the easier and faster it will be transported into a cell. In medical science for example, this passive transport is crucial for the effe...

The contribution of membrane proteins to membrane diffusion of CO2 in biological systems has been questioned for two major reasons: the violation against the Meyer-Overton rule and the disregarding of unstirred layer effects. According to Meyer-Overtons rule membrane diffusion of gasses is not limited by the membrane itself nor can it be improved b...

The membrane CO(2) flux into Arabidopsis mesophyll cells was studied using a scanning pH microelectrode. Arabidopsis thaliana mesophyll cells were exposed to photosynthesis-triggering light intensities, which induced cellular CO(2) uptake. Data obtained on a AtPIP1;2 T-DNA insertion line indicated that under these conditions, cellular CO(2) transpo...

Terfezia claveryi is a hypogeous mycorrhizal fungus belonging to the so-called "desert truffles," with a good record as an edible fungus and of considerable economic importance. T. claveryi improves the tolerance to water stress of the host plant Helianthemum almeriense, for which, in field conditions, symbiosis with T. claveryi is valuable for its...

Cellular exchange of carbon dioxide (CO₂) is of extraordinary importance for life. Despite this significance, its molecular mechanisms are still unclear and a matter of controversy. In contrast to other living organisms, plants are physiologically limited by the availability of CO₂. In most plants, net photosynthesis is directly dependent on CO₂ di...

Heterologous expression in yeast cells revealed that NtAQP1, a member of the so-called PIP1 aquaporin subfamily, did not display increased water transport activity in comparison with controls. Instead, an increased CO(2)-triggered intracellular acidification was observed. NtPIP2;1, which belongs to the PIP2 subfamily of plant aquaporins, behaved as...

Plant infestation with parasitic weeds like Cuscuta reflexa induces morphological as well as biochemical changes in the host and the parasite. These modifications could be caused by a change in protein or gene activity. Using a comparative macroarray approach Cuscuta genes specifically upregulated at the host attachment site were identified. One of...

The continuous progress in the structural and functional characterization of aquaporins increasingly attracts attention to study their roles in certain mammalian diseases. Although several structures of aquaporins have already been solved by crystallization, the challenge of producing sufficient amounts of functional proteins still remains. CF (cel...

The plant parasite Cuscuta reflexa induces various responses in compatible and incompatible host plants. The visual reactions of both types of host plants including obvious morphological changes require the recognition of Cuscuta ssp. A consequently initiated signaling cascade is triggered which leads to a tolerance of the infection or, in the case...

Cuscuta reflexa induces a variety of reaction in its hosts. Some of these are visual reactions, and it is clear that these morphological changes are preceded by events at the molecular level, where signal transduction is one of the early processes. Calcium (Ca(2+)) release is the major second messenger during signal transduction, and we therefore s...

CO(2) faces a series of resistances while diffusing between the substomatal cavities and the sites of carboxylation within chloroplasts. The absence of techniques to measure the resistance of individual steps makes it difficult to define their relative importance. Resistance to diffusion through intercellular airspace differs between leaves, but is...

The majority of plants are unable to evade unfavorable conditions such as flooding, salinity, or drought. Therefore, a fine-tuned water homeostasis appears to be of crucial importance for plant survival, and it was assumed that aquaporins play a significant role in these processes. Regulation of plant aquaporin conductivity was suggested to be achi...

Photosynthesis is often limited by the rate of CO(2) diffusion from the atmosphere to the chloroplast. The primary resistances for CO(2) diffusion are thought to be at the stomata and at photosynthesizing cells via a combination resulting from resistances of aqueous solution as well as the plasma membrane and both outer and inner chloroplast membra...

The impact of aquaporin function on plant water balance is discussed. The significance of these proteins for root water uptake, water conductance in the xylem, including embolism refilling and the role of plant aquaporins in leaf physiology, is described. Emphasis is placed on certain aspects of water stress reactions and the correlation of aquapor...

Background Plant leaf movements can be mediated by specialized motor organs, the pulvini, or can be epinastic (i.e. based on different growth velocities of the adaxial and abaxial halves of the leaf). Both processes are associated with diurnally regulated increases in rates of membrane water transport, which in many cases has been shown to be facil...

Cuscuta spp represent a unique group of holoparasitic dicotyledonous plants which can infect nearly all dicotyledonous species. Lacking roots and leaves, these parasitic plants are completely dependent on nutrients, carbohydrates and water from host plants. The physical connection between parasite and host is mediated by specific organs, called hau...

Functional analysis of heterologously expressed TaTIP2; 2 by means of stopped-flow spectrometric studies provide evidence for water and ammonia conductivity. A series of experiments under increasing pH indicate that the gaseous NH3, rather than the ammonium ion NH4+ was transported. Results from inhibitor studies strongly suggest that NH3 is not tr...

The effects of short-term (minutes) variations of CO2 concentration on mesophyll conductance to CO2 (gm) were evaluated in six different C3 species by simultaneous measurements of gas exchange, chlorophyll fluorescence, online carbon isotope discrimination and a novel curve-fitting method. Depending on the species, gm varied from five- to ninefold,...

Sole ammonium supply provokes negative effects on dry-mass formation, leaf growth, and water uptake of ammonium-sensitive plants. To study the effects of N form on nutrient and water uptake and aquaporin expression, French bean plants were grown in a split-root system. Five treatments were compared: homogeneous nitrate (NN) and ammonium (AA) supply...

Plants have been reported to contain a large set of aquaporins (38 for Arabidopsis), which has been divided into four subfamilies on the basis of similarities in their amino acid sequences. They belong to the large superfamily of major intrinsic proteins (MIP), which was the basis for the nomenclature PIP, TIP, and NIP, also indicating the subcellu...

The measurement of the response of net photosynthesis to leaf internal CO2 (i.e. A–Ci curves) is widely used for ecophysiological studies. Most studies did not consider CO2 exchange between the chamber and the surrounding air, especially at the two extremes of A–Ci curves, where large CO2 gradients are created, leading to erroneous estimations of A...

Almost all land plants have developed a symbiosis with arbuscular mycorrhizal fungi. Establishment of the association is accompanied by structural changes in the plant root. During arbuscule formation fungal hyphae penetrate the root apoplast and install highly specialized interfaces for solute transport between plant and fungus. The periarbuscular...

To study the molecular bases of water transport in olive we characterized cDNAs from Olea europaea cv "Leccino" related to the aquaporin (AQP) gene family. A phylogenetic analysis of the corresponding polypeptides confirmed that they were part of water channel proteins localized in the plasma membrane and in the tonoplast. The full-length sequences...

Dodder or Cuscutaceae are holoparasitic plants subsisting on other dicotyledonous plants. The infection process is initiated by adherence of Cuscuta prehaustoria to the host surface, followed by penetration attempts by hyphae. In the case of a successful infection, these organs connect the parasite's vascular tissue to that of the host. Here we sho...

Leaf mesophyll conductance to CO(2) (g(m)) has been recognized to be finite and variable, rapidly adapting to environmental conditions. The physiological basis for fast changes in g(m) is poorly understood, but current reports suggest the involvement of protein-facilitated CO(2) diffusion across cell membranes. A good candidate for this could be th...

Due to the fact that most plants are immobile, a rapid response of physiological processes to changing environmental conditions is essential for their survival. Thus, in comparison to many other organisms, plants might need a more sophisticated tuning of water balance. Among others, this is reflected by the comparable large amount of aquaporin gene...

Aquaporins are membrane-intrinsic proteins that facilitate membrane transport of water and small solutes or even gases. Aquaporin genes are found in almost all living organisms. In plants the proteins account for water uptake and transport as well as CO2 availability for photosynthesis. These processes are subjected to diurnal or circadian regimes....

Although very often exposed to a rapid changing environment, plants are in general unable to evade from unfavourable conditions. Therefore, a fine tuned adaptation of physiology including the water balance appears to be of crucial importance. As a consequence a relatively large number of aquaporin genes are present in plant genomes. So far aquapori...

Strawberry (Fragaria x ananassa) fruit accumulate (hydroxy)cinnamoyl glucose (Glc) esters, which may serve as the biogenetic precursors of diverse secondary metabolites, such as the flavor constituents methyl cinnamate and ethyl cinnamate. Here, we report on the isolation of a cDNA encoding a UDP-Glc:cinnamate glucosyltransferase (Fragaria x ananas...

Water is the major component of all living cells, and efficient regulation of water homeostasis is essential for many biological processes. The mechanism by which water passes through biological membranes was a matter of debate until the discovery of the aquaporin water channels. Aquaporins are intrinsic membrane proteins characterized by six trans...

The formation of ectomycorrhizas, a tight association between fine roots of trees and certain soil fungi, improves plant nutrition in a nutrient-limited environment and may increase plant survival under water stress conditions. To investigate the impact of mycorrhiza formation on plant water uptake, seven genes coding for putative water channel pro...

We investigated in two tobacco (Nicotiana tabacum) plant lines (wildtype or antisense mutant) whether impairment in expression of the plasma membrane aquaporin gene (NtAQP1) affects the arbuscular mycorrhizal (AM) fungal colonisation pattern or the symbiotic efficiency of AM fungi. These two objectives were investigated under well-watered and droug...

Several processes during sexual reproduction in higher plants involve the movement of water between cells or tissues, such as occurs during dehiscence of the anther and hydration of the pollen grain after it is deposited on a stigma. To get more insight in these processes, a set of putative aquaporins was cloned and it was found that at least 15 ar...

A suppressive subtractive hybridization technique was used to identify genes, which were induced during the early phases of the interaction between dodder (Cuscuta reflexa), a phanerogamic parasite, and its incompatible host plant tomato. One of the identified genes encodes a tomato xyloglucan endotransglycosylase/hydrolase (XTH)--an enzyme involve...

Epinastic leaf movement of tobacco is based on differential growth of the upper and lower leaf surface and is distinct from the motor organ-driven mechanism of nyctinastic leaf movement of, for example, mimosa species. The epinastic leaf movement of tobacco is observed not only under diurnal light regimes but also in continuous light, indicating a...

Several processes during sexual reproduction in higher plants involve the movement of water between cells or tissues, such as occurs during dehiscence of the anther and hydration of the pollen grain after it is deposited on a stigma. To get more insight in these processes, a set of putative aquaporins was cloned and it was found that at least 15 ar...

In a previous paper, we showed that nitrate reductase (NR; EC 1.6.6.1) from leaves of Ricinus communis L. differed from most other higher-plant NRs by an unusually strong Mg2+-sensitivity, a different pH-activity profile and only little ATP-dependent inactivation [A. Kandlbinder et al. (2000) J Exp Bot 51:1099-1105]. In order to elucidate these dev...

Aquaporins, found in virtually all living organisms, are membrane-intrinsic proteins that form water-permeable complexes. The mammalian aquaporin AQP1 has also shown CO2 permeability when expressed heterologously in Xenopus oocytes, although whether this is a biochemical curiosity or of physiological significance is a matter of debate. Here we repo...

Abundant flavin binding sites have been found in membranes of plants and fungi. With flavin mononucleotide-agarose affinity columns, riboflavin-binding activity from microsomes of Cucurbita pepoL. hypocotyls was purified and identified as a specific PIP1-homologous protein of the aquaporin family. Sequences such as gi|2149955 in Phaseolus vulgaris,...

IntroductionDiversity of Species with Aquaporin GenesMicrobial AquaporinsStructural Properties of AquaporinsFunctional Analysis of AquaporinsUnspecific AquaporinsComplexity of Microbial MIP-like Channel GenesGene StructuresPhysiological Indications for Protein-mediated Membrane Water TransportThe Human Aquaporin 1 as a ModelThe Escherichia coli Aqu...

Strawberry fruits contain an uncommon group of key aroma compounds with a 2,5-dimethyl-3(2H)-furanone structure. Here, we report on the methylation of 2,5-dimethyl-4-hydroxy-3(2H)-furanone (DMHF) to 2,5-dimethyl-4-methoxy-3(2H)-furanone (DMMF) by a S-adenosyl-L-methionine dependent O-methyltransferase, the cloning of the corresponding cDNA and char...

The molecular functions of several aquaporins are well characterized (e.g., by analysis of aquaporin-expressing Xenopus oocytes). However, their significance in the physiology of water transport in multicellular organisms remains uncertain. The tobacco plasma membrane aquaporin NtAQP1 was used to elucidate this issue. By comparing antisense plants...

Leaf-moving organs, remarkable for the rhythmic volume changes of their motor cells, served as a model system in which to study the regulation of membrane water fluxes. Two plasma membrane intrinsic protein homolog genes, SsAQP1 and SsAQP2, were cloned from these organs and characterized as aquaporins in Xenopus laevis oocytes. Osmotic water permea...

SummaryA new aquaporin from Nicotiana tabacum (cv. Samsun) was characterized. It shares sequence homology to the Arabidopsis thaliana PIP1 protein family. By two-phase partitioning and immunoblot analysis, plasma membrane localization could be demonstrated. The corresponding mRNA is highly abundant in roots and flowers, while it is rarely expressed...

Young maize seedlings (Zea mays L. cv.Giza 2) were exposed to moderate salinity in hydroponic culture. NADH-nitrate reductase (NR) activity (E.C. 1·6.6·1), NR activation state, NR-mRNA-steady state levels and major solute contents in leaves and roots were investigated. With increasing external salt concentration, Na+, Cl−, sugars, amino acids and q...

This paper gives a summary of a project to characterize a tobacco aquaporin. The cDNA and gene, including the 5′ upstream region, for the tobacco aquaporin NtAQP1 has been isolated and the encoded protein characterized. The significance of promoter regions for an abscisic acid‐ and gibberellic acid‐induced gene expression could be restricted to a r...

A subtractive suppression hybridization technique was used to identify genes that were induced during early phases of the interaction between Cuscuta reflexa, a phanerogamic plant parasite and the incompatible host tomato (Lycopersicon esculentum Mill.). One of the identified genes encodes a new aquaporin (LeAqp2) from tomato. Its function was conc...

Various Arabidopsis thaliana mutants with defects in phytohormone signal transduction or the reception of light were analysed with regard to their stomatal response in a red, red/blue light irradiation programme. Stomatal response to light was detected with a customized gas exchange measurement device, optimized for the small model plant. Small tra...

Various Arabidopsis thaliana mutants with defects in phytohormone signal transduction or the reception of light were analysed with regard to their stomatal response in a red, red/blue light irradiation programme. Stomatal response to light was detected with a customized gas exchange measurement device, optimized for the small model plant. Small tra...

 The aquaporin NtAQP1 from Nicotiana tabacum L. is insensitive to heavy-metal ions. In addition to water, the transport of urea or glycerol is facilitated by this plasma-membrane-located water channel. Northern hybridization and whole-mount in situ hybridization revealed a high steady-state level of NtAQP1-RNA in roots, a decreased content in shoot...

 The genome of the model legume Medicago truncatula Gaertn. was screened for the presence of genes encoding tonoplast intrinsic proteins, and a gene family was identified. The cDNA fragments of two members of the multigene family were cloned from roots inoculated with an arbuscular mycorrhizal fungus. Transcript accumulation in roots could be detec...

Young maize seedlings (Zea mays L. cv.Giza 2) were exposed to moderate salinity in hydroponic culture. NADH-nitrate reductase (NR) activity (E.C. 1.6.6.1), NR activation state, NR-mRNA-steady state levels and major solute contents in leaves and roots were investigated. With increasing external salt concentration, Na+, Cl-, sugars, amino acids and q...

The characterization of a plant MIP from tobacco (Nicotiana tabacum) regarding its function as an aquaporin and role in the plant will be presented.

Membrane intrinsic proteins (MIPs) that facilitate water transport across biological membranes have been identified in many single- and multi-cellular organisms. Starting from an overview about the MIP protein family, common structures of aquaporins are described. A detailed report about tonoplast- and plasma-membrane-located plant aquaporins is gi...

Recent findings concerning the sensitivity of aquaporins to heavy metal ions and their possible consequences to plant physiology are discussed. Furthermore, results obtained from heterologous expression of plant aquaporin cRNAs in Xenopus oocytes are related to the situation in the plant. An assessment of the significance of water flux regulation b...

A new aquaporin from Nicotiana tabacum (cv. Samsun) was characterized. It shares sequence homology to the Arabidopsis thaliana PIP1 protein family. By two-phase partitioning and immunoblot analysis, plasma membrane localization could be demonstrated. The corresponding mRNA is highly abundant in roots and flowers, while it is rarely expressed in lea...

Water uptake ofArabidopsis thaliana protoplasts was measured after transfer into hypo-osmotic conditions. The time-dependent swelling of protoplast populations was monitored by a Coulter counter device. In order to ascertain the contribution of the plasma membrane intrinsic protein 1b (PIP1b) to the membrane's water permeability, protoplasts of fiv...

The plasmalemma ofArabidopsis thaliana contains several different aquaporins. These members of the major-intrinsic-protein family are permeable and selective for water. The apparent redundancy of aquaporins might be explained by tissue-specific expression of the corresponding gene. In order to investigate and compare the expression levels of five g...