Mauricio Hunsche

Stomatal penetration by aqueous solutions – an update involving leaf surface particles

The recent visualization of stomatal nanoparticle uptake ended a 40-yr-old paradigm. Assuming clean, hydrophobic leaf surfaces, the paradigm considered stomatal liquid water transport to be impossible as a result of water surface tension. However, real leaves are not clean, and deposited aerosols may change hydrophobicity and water surface tension. Droplets containing NaCl, NaClO(3), (NH(4))(2) SO(4), glyphosate, an organosilicone surfactant or various combinations thereof were evaporated on stomatous abaxial and astomatous adaxial surfaces of apple (Malus domestica) leaves. The effects on photosynthesis, necrosis and biomass were determined. Observed using an environmental scanning electron microscope, NaCl and NaClO(3) crystals on hydrophobic tomato (Solanum lycopersicum) cuticles underwent several humidity cycles, causing repeated deliquescence and efflorescence of the salts. All physiological parameters were more strongly affected by abaxial than adaxial treatments. Spatial expansion and dendritic crystallization of the salts occurred and cuticular hydrophobicity was decreased more rapidly by NaClO(3) than NaCl. The results confirmed the stomatal uptake of aqueous solutions. Humidity fluctuations promote the spatial expansion of salts into the stomata. The ion-specific effects point to the Hofmeister series: chaotropic ions reduce surface tension, probably contributing to the defoliant action of NaClO(3), whereas the salt spray tolerance of coastal plants is probably linked to the kosmotropic nature of chloride ions.

“Breath figures” on leaf surfaces—formation and effects of microscopic leaf wetness

“Microscopic leaf wetness” means minute amounts of persistent liquid water on leaf surfaces which are invisible to the naked eye. The water is mainly maintained by transpired water vapor condensing onto the leaf surface and to attached leaf surface particles. With an estimated average thickness of less than 1 μm, microscopic leaf wetness is about two orders of magnitude thinner than morning dewfall. The most important physical processes which reduce the saturation vapor pressure and promote condensation are cuticular absorption and the deliquescence of hygroscopic leaf surface particles. Deliquescent salts form highly concentrated solutions. Depending on the type and concentration of the dissolved ions, the physicochemical properties of microscopic leaf wetness can be considerably different from those of pure water. Microscopic leaf wetness can form continuous thin layers on hydrophobic leaf surfaces and in specific cases can act similar to surfactants, enabling a strong potential influence on the foliar exchange of ions. Microscopic leaf wetness can also enhance the dissolution, the emission, and the reaction of specific atmospheric trace gases e.g., ammonia, SO2, or ozone, leading to a strong potential role for microscopic leaf wetness in plant/atmosphere interaction. Due to its difficult detection, there is little knowledge about the occurrence and the properties of microscopic leaf wetness. However, based on the existing evidence and on physicochemical reasoning it can be hypothesized that microscopic leaf wetness occurs on almost any plant worldwide and often permanently, and that it significantly influences the exchange processes of the leaf surface with its neighboring compartments, i.e., the plant interior and the atmosphere. The omission of microscopic water in general leaf wetness concepts has caused far-reaching, misleading conclusions in the past.

Investigation of Leaf Diseases and Estimation of Chlorophyll Concentration in Seven Barley Varieties Using Fluorescence and Hyperspectral Indices

Leaf diseases, such as powdery mildew and leaf rust, frequently infect barley plants and severely affect the economic value of malting barley. Early detection of barley diseases would facilitate the timely application of fungicides. In a field experiment, we investigated the performance of fluorescence and reflectance indices on (1) detecting barley disease risks when no fungicide is applied and (2) estimating leaf chlorophyll concentration (LCC). Leaf fluorescence and canopy reflectance were weekly measured by a portable fluorescence sensor and spectroradiometer, respectively. Results showed that vegetation indices recorded at canopy level performed well for the early detection of slightly-diseased plants. The combined reflectance index, MCARI/TCARI, yielded the best discrimination between healthy and diseased plants across seven barley varieties. The blue to far-red fluorescence ratio (BFRR_UV) and OSAVI were the best fluorescence and reflectance indices for estimating LCC, respectively, yielding R 2 of 0.72 and 0.79. Partial

Surfactant-induced deposit structures in relation to the biological efficacy of glyphosate on easy- and difficult-to-wet weed species

BACKGROUND Typical active ingredient (AI) residue patterns are formed during droplet drying on plant surfaces owing to the interaction of spray solution characteristics and leaf micromorphology. Currently, comparatively little is known about the influence of AI deposit patterns within a spray droplet residue area on the penetration and biological efficacy of glyphosate. A scanning electron microscope with energy dispersive X-ray microanalysis has been used to characterise residue patterns and to quantify the area ultimately covered by glyphosate within the droplet spread area. RESULTS The easy-to-wet weed species Stellaria media L. and Viola arvensis L., as well as the difficult-to-wet Chenopodium album L. and Setaria viridis L., differing in their surface micromorphology, have been used. Rapeseed oil ethoxylates (RSO 5 or RSO 60) were added to glyphosate solutions to provide different droplet spread areas. Addition of RSO 5 enhanced droplet spread area more than RSO 60, and both caused distinct glyphosate residue patterns. The biological efficacy of treatment solutions showed no significant correlation with the area ultimately covered by glyphosate. CONCLUSION The results have implications on herbicide uptake models. This study shows that droplet spread area does not correspond to the area ultimately covered by glyphosate, and that the latter does not affect glyphosate phytotoxicity.

Ecologically relevant UV-B dose combined with high PAR intensity distinctly affect plant growth and accumulation of secondary metabolites in leaves of Centella asiatica L. Urban

We investigated the effects of environmentally relevant dose of ultraviolet (UV)-B and photosynthetic active radiation (PAR) on saponin accumulation in leaves on the example of Centella asiatica L. Urban. For this purpose, plants were exposed to one of four light regimes i.e., two PAR intensities with or without UV-B radiation. The experiment was conducted in technically complex sun simulators under almost natural irradiance and climatic conditions. As observed, UV-B radiation increased herb and leaf production as well as the content of epidermal flavonols, which was monitored by non-destructive fluorescence measurements. Specific fluorescence indices also indicate an increase in the content of anthocyanins under high PAR; this increase was likewise observed for the saponin concentrations. In contrast, UV-B radiation had no distinct effects on saponin and sapogenin concentrations. Our findings suggest that besides flavonoids, also saponins were accumulated under high PAR protecting the plant from oxidative damage. Furthermore, glycosylation of sapogenins seems to be important either for the protective function and/or for compartmentalization of the compounds. Moreover, our study revealed that younger leaves contain higher amounts of saponins, while in older leaves the sapogenins were the most abundant constituents. Concluding, our results proof that ambient dose of UV-B and high PAR intensity distinctly affect the accumulation of flavonoids and saponins, enabling the plant tissue to adapt to the light conditions.

Centelloside accumulation in leaves of Centella asiatica is determined by resource partitioning between primary and secondary metabolism while influenced by supply levels of either nitrogen, phosphorus or potassium.

In the present study we aimed to investigate the relevance of either N, P or K supply for herb and leaf yield and for centelloside concentrations in Centella asiatica L. Urban leaves. In this regard, we elucidated the causal relationship between assimilation rate, leaf N, P and K concentrations, herb and leaf production, and centelloside accumulation. The experiments were conducted consecutively in a greenhouse where C. asiatica was grown in hydroponic culture and fertigated with nutrient solutions at either 0, 30, 60, 100 or 150% of the N, P or K amount in a standard Hoagland solution. In general, the increase in N, P or K supply enhanced assimilation rate and herb and leaf yield. However, exceeding specific thresholds, the high availability of one single nutrient caused lower leaf N concentrations and a decline in assimilation rate and plant growth. Irrespective of N, P and K supply, the leaf centelloside concentrations were negatively associated with herb and leaf yield, which is in accordance with the assumptions of the carbon/nutrient balance and the growth differentiation balance hypotheses. Moreover, we found strong negative correlations between saponins and leaf N concentrations, while the respective sapogenins were negatively correlated with K concentrations. Using C. asiatica as model system, our experiments reveal for the first time that the accumulation of saponins and sapogenins is affected by resource allocation between primary and secondary metabolism and that besides carbon, also nutrient availability is relevant for the regulation of the centelloside synthesis. Finally, our results highlight the huge potential of optimized and carefully controlled mineral nutrition of medicinal plants for steering the bio-production of high-quality natural products.

Effects of NaCl on surface properties, chlorophyll fluorescence and light remission, and cellular compounds of Grewia tenax (Forssk.) Fiori and Tamarindus indica L. leaves

Seedlings of the salt-tolerant plant grewia [Grewia tenax (Forssk.) Fiori] and the moderately salt-tolerant tamarind (Tamarindus indica L.) were grown under controlled conditions and treated daily with NaCl solutions to investigate mechanisms of tolerance to salinity. Leaf micromorphology, cuticular wax load, chlorophyll fluorescence and light remission, as well as antioxidative potential were evaluated. As confirmed by energy-dispersive X-ray microanalysis in both species, absorption of sodium and chlorine increased with rising NaCl concentration in the treatment solution. In parallel, accumulation of calcium in grewia leaves was strongly reduced, leading to less crystals of calcium oxalate in leaf tissue. In grewia the cuticular wax load, chlorophyll content, and electron transport rate (ETR) were significantly reduced by comparatively low NaCl concentrations. In tamarind, in contrast, wax load and ETR were not significantly affected, while the decrease of chlorophyll content was less pronounced. Measurements of the antioxidative capacity and the imbalance between values of lipophilic and hydrophilic extracts at different NaCl concentrations confirmed that grewia is more salt tolerant than tamarind. This higher tolerance degree seemed to be associated with grewias’ more efficient scavenging of free radicals and the regulation of the antioxidative potential in lipophilic and hydrophilic extracts.

Sensing drought- and salinity-imposed stresses on tomato leaves by means of fluorescence techniques

In our study, we investigated whether multiple fluorescence indices may be used to sense physiological changes in tomato plants (Solanum lycopersicum L.) caused by salinity and water deficit as single or combined stresses. The fluorescence intensity in the blue (B), red (R) and far-red (FR) spectral regions and the pulse-amplitude-modulated (PAM) chlorophyll fluorescence, were recorded on a weekly basis in the scope of a long-term experiment. The results indicate the coefficient of photochemical quenching (qL), the B to FR fluorescence ratio and the logarithm of the FR fluorescence ratio after R and UV-light excitation as appropriate parameters to sense the response of plants to the imposed stress. The qL revealed the impact of water deficiency, whereas the two multispectral ratios revealed the influence of combined salinity and water shortage. Despite minor changes in the chlorophyll concentration, salinity and water deficit, when combined, had an additive impact on the chlorophyll fluorescence. Overall, the fluorescence signals of ‘Rio Grande’ were more affected by the induced stresses compared to ‘Harzfeuer’. The multiparametric fluorescence technique, confirming the trends obtained with the PAM-method, reveals promising perspectives for the ‘in situ’ evaluation of the physiological status of horticultural crops.

Selected Calcium Salt Formulations: Interactions between Spray Deposit Characteristics and Ca Penetration with Consequences for Rain-Induced Wash-Off

ABSTRACT The effect of two ethoxylated rapeseed oils (RSO) as surfactants for foliar-applied calcium chloride CaCl2 and calcium acetate Ca(CH3COO)2 on deposit characteristics, penetration, and calcium (Ca) residues after rainfall was investigated in columnar apple trees (CAT, Malus domestica Borkh.) and apple seedlings. Irrespective of salt and apple variety, both surfactants enhanced droplet spreading and led to a greater area covered by calcium within the droplet residues. The addition of the rather lipophilic surfactant RSO 5 and the rather hydrophilic RSO 60 increased penetration of Ca from both salts through isolated apple leaf cuticular membranes. Our data demonstrate that penetration is better reflected by area covered with calcium than by droplet spreading area. Considering the point of deliquescence (POD) from the salts and results of cuticular Ca penetration, we postulate that in case of CaCl2 reduction of wash-off was due to enhanced penetration, whereas for Ca(CH3COO)2 wash-off was decreased due to distinct enhancement of rainfastness.

Presymptomatic Detection of Powdery Mildew Infection in Winter Wheat Cultivars by Laser-Induced Fluorescence

The sensor-based monitoring of diseases under controlled conditions establishes an objective tool that allows a better understanding of the pathogen–plant interactions in different situations. The purpose of our work was to implement the presymptomatic detection of powdery mildew on wheat leaves shortly after fungus inoculation by spectral and time-resolved laser-induced fluorescence spectroscopy. In the general scope of plant phenotyping, we hypothesized that it is possible to discriminate between wheat genotypes that are either resistant or susceptible to powdery mildew. According to our results, the presymptomatic detection of powdery mildew on wheat leaves was accomplished, irrespective of genotype, as early as one day after inoculation using the fluorescence amplitude ratio F451:F522. Similarly, the ratios F451:F522, F522:F687, and F522:F736 of the half-bandwidth are also appropriate parameters. Furthermore, in the spectral range between 410 nm and 620 nm, the mean lifetime was significantly longer in inoculated leaves than it was in control leaves. Finally, the short-term (10–12 hour) increase of the fluorescence mean lifetime at 530 nm and 560 nm following the inoculation suggests that the speed of the plant reaction might be associated to its resistance to the pathogen. Based on this information, we conclude that determinations of ultraviolet, laser-induced fluorescence intensity and lifetime are suitable approaches to presymptomatically detect powdery mildew on wheat leaves one day after inoculation.