Glutathione Modulation in PVYNTN Susceptible and Resistant Potato Plant Interactions.

Glutathione is a metabolite that plays an important role in plant response to biotic stress through its ability to remove reactive oxygen species, thereby limiting the degree of potential oxidative damage. It can couple changes in the intracellular redox state to the development, especially the defense responses, of plants. Several studies have focused on measuring glutathione levels in virus infected plants, but have not provided complete information. Therefore, we analyzed, for the first time, the content of glutathione as well as its ultrastructural distribution related to susceptible and hypersensitive potato-Potato virus Y NTN (PVYNTN) interaction, with an aim of providing new insight into interactive responses to PVYNTN stress. Our findings reported that the inoculation of PVYNTN caused a dynamic increase in the content of glutathione, not only in resistance but also in susceptible reaction, especially at the first steps of plant-virus interaction. Moreover, the increase in hypersensitive response was much more dynamic, and accompanied by a significant reduction in the content of PVYNTN. By contrast, in susceptible potato Irys, the content of glutathione decreased between 7 and 21 days after virus inoculation, which led to a significant increase in PVYNTN concentration. Additionally, our findings clearly indicated the steady induction of two selected potato glutathione S-transferase StGSTF1 and StGSTF2 genes after PVYNTN inoculation, regardless of the interaction type. However, the relative expression level of StGSTF1 did not significantly differ between resistant and susceptible plants, whereas the relative expression levels of StGSTF2 differed between susceptible and resistant reactions. Therefore, we proposed that StGSTF2 can act as a marker of the type of response to PVYNTN. Our observations indicated that glutathione is an important component of signaling as well as the regulatory network in the PVYNTN-potato pathosystem. In resistance responses to PVYNTN, this metabolite activates plant defenses by reducing potential damage to the host plant cell, causing a reduction in virus concentration, while it can also be involved in the development of PVYNTN elicited symptoms, as well as limiting oxidative stress, leading to systemic infection in susceptible potato plants.

Effect of AuNPs and AgNPs on the Antioxidant System and Antioxidant Activity of Lavender (Lavandula angustifolia Mill.) from In Vitro Cultures.

The aim of this study was to determine the effect of gold and silver nanoparticles on the activity of antioxidant enzymes (ascorbate peroxidase (APX), superoxide dismutase (SOD), guaiacol peroxidase (POX), and catalase (CAT)), the free radical scavenging capacity, and the total polyphenol capacity of lavender (Lavandula angustifolia Mill.) cultivar "Munstead" propagated in vitro. In the experiment, fragments of lavender plants were cultivated in vitro on medium with the addition of 1, 2, 5, 10, 20, and 50 mg∙dm-3 of AgNPs or AuNPs (particle sizes 24.2 ± 2.4 and 27.5 ± 4.8 nm, respectively). It was found that the nanoparticles increase the activity of the antioxidant enzymes APX and SOD; however, the reaction depends on the NP concentration. The highest APX activity is found in plants propagated on media with 2 and 5 mg∙dm-3 of AgNPs. AuNPs significantly increase the APX activity when added to media with a concentration of 10 mg∙dm-3. The highest SOD activity is recorded at 2 and 5 mg∙dm-3 AgNP and AuNP concentrations. The addition of higher concentrations of nanoparticles to culture media results in a decrease in the APX and SOD activity. The addition of AuNPs to culture media at concentrations from 2 to 50 mg∙dm-3 increases the POX activity in comparison to its activity when AgNPs are added to the culture media. No significant influence of NPs on the increase in CAT activity was demonstrated. AgNPs and AuNPs increased the free radical scavenging capacity (ABTS•+). The addition of NPs at concentrations of 2 and 5 mg∙dm-3 increased the production of polyphenols; however, in lower concentrations it decreased their content in lavender tissues.

Antioxidant properties of potato tubers (Solanum tuberosum L.) as a consequence of genetic potential and growing conditions.

The concentrations of the bioactive compounds in potato tubers are determined by both genetic potential and environmental factors. The purpose of the experiment was to determine the influence of organic and integrated production on the expression of the genetic potential with respect to the antioxidant properties of potato tubers and to evaluate its stability under different environmental conditions. This phenotyping was performed on seven new breeding lines (tetraploid clones) and three modern cultivars: Jelly, Satina and Tajfun. The results indicated that production system and location significantly influenced the antioxidant capacity measured by FRAP method. Organic farming and the location Chwalowice were characterized by higher values. Furthermore, anitioxidative capacity measured by FRAP method was correlated with chlorogenic acid content (r = 0.590**) and glutathione fractions, especially with the reduced form (GSH, r = 0.692**). Multidimensional comparative analysis (MCA) proved a better development of antioxidant properties of potato tubers in the organic cultivation system when compared with the integrated. Especially favorable were growing conditions at Boguchwala (organic) and worst at Mlochów (integrated). From all investigated varieties, the best antioxidant properties were found in 'Satina' and 'Jelly'. Clones TG-97-403 and 13-VIII-45 developed the weakest health promoting traits.

Gas Chromatography-Mass Spectrometry (GC-MS) Analysis of Essential Oils from AgNPs and AuNPs Elicited Lavandula angustifolia in vitro Cultures.

The aim of this study was to determine how the addition of gold and silver nanoparticles to culture media affects the composition of essential oils extracted from Lavandula angustifolia propagated on MS media with the addition of 10 and 50 mg•dm-3 of gold (24.2 ± 2.4 nm) and silver (27.5 ± 4.8 nm) nanocolloids. The oil extracted from the lavender tissues propagated on the medium with 10 mg•dm-3 AgNPs (silver nanoparticles) differed the most with respect to the control; oil-10 compounds were not found at all, and 13 others were detected which were not present in the control oil. The addition of AuNPs (gold nanoparticles) and AgNPs to the media resulted in a decrease of lower molecular weight compounds (e.g., α- and ß-pinene, camphene, δ-3-carene, p-cymene, 1,8-cineole, trans-pinocarveol, camphoriborneol), which were replaced by those of a higher molecular weight (τ- and α-cadinol 9-cedranone, cadalene, α-bisabolol, cis-14-nor-muurol-5-en-4-one, (E,E)-farnesol).

Development of a Sensitive and Specific Polyclonal Antibody for Serological Detection of Clavibacter michiganensis subsp. sepedonicus.

The quarantine bacterium Clavibacter michiganensis subsp. sepedonicus (Cms) causes bacterial ring rot (BRR) in potato but is difficult to detect, hampering the diagnosis of this disease. ELISA immunoassays have not been widely used to detect Cms because commercially available anti-Cms antibodies detect mainly EPS-producing bacteria and can fail to detect strains that do not produce EPS. In the current study, we developed a new type of polyclonal antibody that specifically detects Clavibacter michiganensis subsp. sepedonicus bacteria irrespective of their EPS level. We first found that the presence of bacterial EPS precluded quantitative measurement of bacteria by currently available immunoenzymatic methods, but that washing Cms cells with acidic and basic buffers to remove EPS before analysis successfully standardized ELISA results. We used a mix of three strains of Cms with diverse EPS levels to generate antigen for production of antibodies recognizing Cms cells with and without an EPS layer (IgG-EPS and IgG-N-EPS, respectively). The resulting IgG-N-EPS recognized almost all Cms strains tested in this work regardless of their mucoidal level. The availability of this new antibody renders immunological diagnostics of Cms more sensitive and reliable, as our newly developed antibodies can be used in many type of immunoassays. This work represents an important step forward in efforts to diagnose and prevent the spread of BRR, and the methods and solutions developed in this work are covered by six Polish, one European and one US patents.