Roland Valcke

Photosynthesis of Transgenic Pssu-ipt Tobacco

Summary The elevated content of endogenous cytokinin (CK), as the result of the introduction of the chimeric P ssu-ipt gene in tobacco, induced changes in growth, water regime and photosynthesis of transgenic grafts and rooted plants grown in a greenhouse. Despite closed stomata remarkable water stress was detected in P ssu-ipt plants. The ABA content was lower than in wild type tobacco, indicative of disturbances in ABA transport or metabolism. The rates of net photosynthesis (measured as CO 2 uptake or O 2 evolution) decreased by more than 50% in P ssu-ipt grafts and by 20% in P ssu-ipt rooted plants, respectively. The partial reactions of the electron transport chain in transgenic P ssu-ipt tobacco were differently influenced: the activity of the reaction centre of PSII was hardly affected; the PS I activity and the intersystem electron transport chain was inhibited up to 70%, particularly in the P ssu-ipt grafts. Although the slight decrease in the potential photochemical efficiency of PSII, expressed as F v /F m , was found in transgenic plants, the actual quantum yields, photochemical efficiencies, and qP under steady-state conditions indicated that transgenic plants were not seriously limited by the redox state of QA. The reaction centre of PSII was well preserved. In transgenic grafts, photophosphorylation capacity was strongly reduced, which could correspond with lower non-photochemical quenching. The above mentioned changes are the results of elevated CK content per se rather than the effect of altered water relations in the plants, caused by the disproportion of shoots and root system in both P ssu-ipt grafts and plants.

Imaging viral infection: studies on Nicotiana benthamiana plants infected with the pepper mild mottle tobamovirus

We have studied by kinetic Chl-fluorescence imaging (Chl-FI) Nicotiana benthamiana plants infected with the Italian strain of the pepper mild mottle tobamovirus (PMMoV-I). We have mapped leaf photosynthesis at different points of the fluorescence induction curve as well as at different post-infection times. Images of different fluorescence parameters were obtained to investigate which one could discriminate control from infected leaves in the absence of symptoms. The non-photochemical quenching (NPQ) of excess energy in photosystem II (PSII) seems to be the most adequate chlorophyll fluorescence parameter to assess the effect of tobamoviral infection on the chloroplast. Non-symptomatic mature leaves from inoculated plants displayed a very characteristic time-varying NPQ pattern. In addition, a correlation between NPQ amplification and virus localization by tissue-print was found, suggesting that an increase in the local NPQ values is associated with the areas invaded by the pathogen. Changes in chloroplast ultrastructure in non-symptomatic leaf areas showing different NPQ levels were also investigated. A gradient of ultrastructural modifications was observed among the different areas.

Upward and downward solar-induced chlorophyll fluorescence yield indices of four tree species as indicators of traffic pollution in Valencia.

Passive steady-state chlorophyll fluorescence (Fs) provides a direct diagnosis of the functional status of vegetation photosynthesis. With the prospect of mapping Fs using remote sensing techniques, field measurements are mandatory to understand to which extent Fs allows detecting plant stress in different environments. Trees of four common species in Valencia were classified in either a low or a high local traffic exposure class based on their leaf magnetic value. Upward and downward hyperspectral fluorescence yield (FY) and indices based on the two Fs peaks (at 687 and 741 nm) were calculated. FY indices of P. canariensis and P. x acerifolia were significantly different between the two traffic exposure classes defined, but not for C. australis nor M. alba. While chlorophyll content could not indicate the difference between low and high traffic exposure, the FY(687)/FY(741) peak ratio increased significantly (p < 0.05) for both leaf sides for the higher traffic exposure class.

The Arabidopsis thaliana RNA Editing Factor SLO2, which Affects the Mitochondrial Electron Transport Chain, Participates in Multiple Stress and Hormone Responses

Recently, we reported that the novel mitochondrial RNA editing factor SLO2 is essential for mitochondrial electron transport, and vital for plant growth through regulation of carbon and energy metabolism. Here, we show that mutation in SLO2 causes hypersensitivity to ABA and insensitivity to ethylene, suggesting a link with stress responses. Indeed, slo2 mutants are hypersensitive to salt and osmotic stress during the germination stage, while adult plants show increased drought and salt tolerance. Moreover, slo2 mutants are more susceptible to Botrytis cinerea infection. An increased expression of nuclear-encoded stress-responsive genes, as well as mitochondrial-encoded NAD genes of complex I and genes of the alternative respiratory pathway, was observed in slo2 mutants, further enhanced by ABA treatment. In addition, H2O2 accumulation and altered amino acid levels were recorded in slo2 mutants. We conclude that SLO2 is required for plant sensitivity to ABA, ethylene, biotic, and abiotic stress. Although two stress-related RNA editing factors were reported very recently, this study demonstrates a unique role of SLO2, and further supports a link between mitochondrial RNA editing events and stress response.

Pathogenicity and infection strategies of the fire blight pathogen Erwinia amylovora in Rosaceae: state of the art.

Plants are host to a large amount of pathogenic bacteria. Fire blight, caused by the bacterium Erwinia amylovora, is an important disease in Rosaceae. Pathogenicity of E. amylovora is greatly influenced by the production of exopolysaccharides, such as amylovoran, and the use of the type III secretion system, which enables bacteria to penetrate host tissue and cause disease. When infection takes place, plants have to rely on the ability of each cell to recognize the pathogen and the signals emanating from the infection site in order to generate several defence mechanisms. These mechanisms consist of physical barriers and the production of antimicrobial components, both in a preformed and an inducible manner. Inducible defence responses are activated upon the recognition of elicitor molecules by plant cell receptors, either derived from invading micro-organisms or from pathogen-induced degradation of plant tissue. This recognition event triggers a signal transduction cascade, leading to a range of defence responses [reactive oxygen species (ROS), plant hormones, secondary metabolites, …] and redeployment of cellular energy in a fast, efficient and multiresponsive manner, which prevents further pathogen ingress. This review highlights the research that has been performed during recent years regarding this specific plant-pathogen interaction between Erwinia amylovora and Rosaceae, with a special emphasis on the pathogenicity and the infection strategy of E. amylovora and the possible defence mechanisms of the plant against this disease.

Chlorophyll a fluorescence as a tool for a study of the Potato virus Y effects on photosynthesis of nontransgenic and transgenic Pssu-ipt tobacco

The effect of Potato virus YNTN (PVY) infection upon photosynthesis was analysed in transgenic Pssu-ipt tobacco overproducing endogenous cytokinins in comparison with control, nontransgenic Nicotiana tabacum plants. The course of the infection from the early to the late stage was monitored by measuring of photosynthetic gas exchange and fast chlorophyll (Chl) a fluorescence induction kinetics. Leaf photosynthesis was also analysed using Chl fluorescence imaging (Chl-FI). From the different fluorescence parameters obtained using Chl-FI, the nonphotochemical quenching (NPQ) proved to be the most useful parameter to assess the effect of PVY infection. On the other hand, Chl-FI was found to be inapplicable for any presymptomatic detection of PVY infection in tobacco. The lower accumulation of the virus was found in transgenic plants and corresponded also with the presence of visible symptoms of PVY infection. The net photosynthetic rate (PN), transpiration rate (E), and stomatal conductance (gs) significantly decreased with the progress of the infection in both control plant types and transgenic rooted plants, while transgenic grafts were much less affected. The analysis of the Chl fluorescence transient revealed higher number of silent dissipative reaction centres, higher nonphotochemical dissipation, and significantly lower performance index, PI(abs), in the healthy transgenic grafts. Chl-FI also confirmed significantly higher NPQ in transgenic grafts.

Effects of heavy metals on the fast chlorophyll fluorescence induction kinetics of photosystem II: a comparative study

The effects of toxic concentrations of Cu, Zn and Cd on the fast induction kinetics of fluorescence from photosystem(PS)II were investigated in a comparative way. The fast fluorescence transient from primary leaves of metal-treated bean plants was studied. During several days after metal application, the time course of the changes induced by the different metals was monitored. The results evidenced not only a different time course of the changes in fluorescence related parameters for the three metals, but also different effects on the fluorescence induction kinetics, which could possibly be linked to different mechanisms of action of the metals

A field study on solar-induced chlorophyll fluorescence and pigment parameters along a vertical canopy gradient of four tree species in an urban environment

To better understand the potential uses of vegetation indices based on the sun-induced upward and downward chlorophyll fluorescence at leaf and at canopy scales, a field study was carried out in the city of Valencia (Spain). Fluorescence yield (FY) indices were derived for trees at different traffic intensity locations and at three canopy heights. This allowed investigating within-tree and between-tree variations of FY indices for four tree species. Several FY indices showed a significant (p < 0.05) and important effect of tree location for the species Morus alba (white mulberry) and Phoenix canariensis (Canary Island date palm). The upward FY parameters of M. alba, and the upward to downward ratios at 687 and 741 nm for both species, were significantly related to tree location. It was found that not the total chlorophyll (Chl) content, but rather the Chl a/b ratio showed the strongest correlations with several of the indices applied. Chl a/b was lowest at the bottom level of the highest traffic intensity location for both species due to an increased Chl b, indicating a larger light harvesting complex related to Photosystem II (LHCII) as a response to limiting light. The leaf deposits from traffic observed at this sampling location possibly led to a shading effect, resulting further in an adaptive response of the photosynthetic system and subsequent difference of FY indices. This study therefore indicated the importance of the size of LHCII on the fluorescence emission, observed under different traffic generated pollution conditions.

Apical dominance in Pssu-ipt-transformed tobacco

In Pssu-ipt-transformed tobacco, apical dominance was released by defoliation of the upper nodes, while the apex remained intact. After defoliation, the concentration of cytokinins (CKs) increased whereas IAA remained constant, evoking an increase in the CK/IAA ratio in the buds. Moreover, defoliation resulted in a tremendous increase in the concentrations of aromatic amines (AAs): tyramine (TYR), phenethylamine (PEA) and an as yet unidentified compound. Although the total aliphatic monoamine and polyamine (PA) concentration remained constant, putrescine (PUT) and spermidine (SPD) concentrations in the axillary buds decreased, whereas the concentration of spermine (SPM) increased. Similar changes in PAs and AAs could be observed in the buds of untransformed SR1 plants after decapitation, whereas defoliation without removal of the apex had no effect. This is the first report on the possible involvement of PAs and AAs in apical dominance.

Evaluation of the photosynthetic activity in transgenic tobacco plants with altered endogenous cytokinin content: lessons from cytokinin

Cytokinin is known to be involved in many processes related to plastid development and function but the exact role of cytokinin in photosynthesis remains elusive. To investigate more profoundly the effects of cytokinin in this process, the photosynthetic activity of transgenic Pssuipt and 35S:CKX1 tobacco (Nicotiana tabacum) plants with respectively elevated and reduced endogenous cytokinin content was evaluated. Pigment analysis indicated that elevated endogenous cytokinin content resulted in increased pigment content. Functional analysis of the photosynthetic apparatus by chlorophyll a fluorescence and in vitro electron transport measurements clearly showed that changing the endogenous cytokinin content affects the activity of the photosynthetic apparatus. Surprisingly, both an increase as well as a decrease in cytokinin content results in a better photosynthetic performance. Quenching analysis revealed that the initial responses of the photosynthetic apparatus on a dark-light transition are not affected by changed cytokinin content. However, it has an effect on the further kinetic behavior. Taken together, we suggest that cytokinins can induce structural changes in the different parts of the electron transport chain as also demonstrated by the in vitro electron transport measurements.